BACKGROUND The poor prognosis and rising incidence of esophageal cancer highlight the need for improved therapeutics that are essential prior to treatment.LCL161 is an SMAC(second mitochondrial activator of caspases)m...BACKGROUND The poor prognosis and rising incidence of esophageal cancer highlight the need for improved therapeutics that are essential prior to treatment.LCL161 is an SMAC(second mitochondrial activator of caspases)mimic and inhibitor of apoptosis protein(IAP)antagonist which exhibits anti-tumor effects and improves the chemical sensitivity of many cancers.AIM To ascertain the effects and mechanisms of the SMAC analog LCL161 on esophageal cancer cells.METHODS MTT assay and TUNEL assay were used to detect cell proliferation and apoptosis,respectively.Western blot analysis was used to study the molecular mechanisms of LCL161-induced death of ECA109 cells.RESULTS LCL161 decreased ECA109 cell proliferation in dose-and time-dependent manner and induced apoptosis of ECA109 cells in a dose-dependent manner.Also,LCL161 induced a significant decrease in the expression of the XIAP and significant increase in the expression of Caspase-3.In addition,Bax increased significantly with increasing concentrations of LCL161,and the relative expression of Bax was significantly different between groups.CONCLUSION These findings support the hypothesis that LCL161 can inhibit proliferation and induce apoptosis in esophageal cancer cells by regulating the expression of IAP family members,suggesting that it has potential to be an effective treatment for esophageal squamous cell carcinoma.展开更多
Studies concerning correlations between pituitary adenomas and cell apoptosis have mainly focused on upstream apoptosis signaling, but seldom on downstream mediators. In the present study, second mitochondria-derived ...Studies concerning correlations between pituitary adenomas and cell apoptosis have mainly focused on upstream apoptosis signaling, but seldom on downstream mediators. In the present study, second mitochondria-derived activator of caspases (Smac), X-linked inhibitor of apoptosis protein (XIAP), and caspase-3 protein were qualitatively analyzed using immunohistochemistry, and quantified by western blot. Smac, XIAP, and caspase-3 mRNA expressions were detected by reverse transcription-PCR. Results showed that XIAP protein and mRNA expressions were greater in the invasive pituitary adenoma group compared with the noninvasive pituitary adenoma group. However, Smac and caspase-3 protein and mRNA expressions were lower in the invasive pituitary adenoma group compared with the noninvasive pituitary adenoma group. In the invasive pituitary adenomas, Smac expression was positively correlated with caspase-3 protein and mRNA expression (Protein: r = 0.55, P 0.01; mRNA: r = 0.50, P 0.01). Smac and caspase-3 expressions were negatively correlated with XIAP protein and mRNA expression (Protein: r = -0.56, -0.64, P 0.01; mRNA: r = -0.69, -0.67, P 0.01). However, no significant differences in correlation among Smac, XIAP, and caspase-3 were detectable in noninvasive pituitary adenomas. These data indicated that high expression of XIAP and low expression of Smac and caspase-3 suppressed cell apoptosis and led to enhanced invasiveness of pituitary adenomas. Thus, Smac, XIAP, and caspase-3 may be useful markers in determining the invasive behavior of pituitary adenomas.展开更多
Wallerian degeneration is a complex biological process that occurs after nerve injury,and involves nerve degeneration and regeneration.Schwann cells play a crucial role in the cellular and molecular events of Walleria...Wallerian degeneration is a complex biological process that occurs after nerve injury,and involves nerve degeneration and regeneration.Schwann cells play a crucial role in the cellular and molecular events of Wallerian degeneration of the peripheral nervous system.However,Wallerian degeneration regulating nerve injury and repair remains largely unknown,especially the early response.We have previously reported some key regulators of Wallerian degeneration after sciatic nerve injury.Baculoviral inhibitor of apoptosis protein repeat-containing protein 3(BIRC3)is an important factor that regulates apoptosis-inhibiting protein.In this study,we established rat models of right sciatic nerve injury.In vitro Schwann cell models were also established and subjected to gene transfection to inhibit and overexpress BIRC3.The data indicated that BIRC3 expression was significantly up-regulated after sciatic nerve injury.Both BIRC3 upregulation and downregulation affected the migration,proliferation and apoptosis of Schwan cells and affected the expression of related factors through activating c-fos and ERK signal pathway.Inhibition of BIRC3 delayed early Wallerian degeneration through inhibiting the apoptosis of Schwann cells after sciatic nerve injury.These findings suggest that BIRC3 plays an important role in peripheral nerve injury repair and regeneration.The study was approved by the Institutional Animal Care and Use Committee of Nantong University,China(approval No.2019-nsfc004)on March 1,2019.展开更多
Apoptosis proteins play an important role in the development and homeostasis of an organism. The elucidation of the subcellular locations and functions of these proteins is helpful for understanding the mechanism of p...Apoptosis proteins play an important role in the development and homeostasis of an organism. The elucidation of the subcellular locations and functions of these proteins is helpful for understanding the mechanism of programmed cell death. In this paper, the recurrent quantification analysis, Hilbert-Huang transform methods, the maximum relevance and minimum redundancy method and support vector machine are used to predict the subcellular location of apoptosis proteins. The validation of the jackknife test suggests that the proposed method can improve the prediction accuracy of the subcellular location of apoptosis proteins and its application may be promising in other fields.展开更多
The expression of X-linked inhibitor of apoptosis protein (XIAP) gene and its effect on chemotherapeutic sensitivity in bladder carcinoma was explored. By using immunohistochemistry, the expression of XIAP was detecte...The expression of X-linked inhibitor of apoptosis protein (XIAP) gene and its effect on chemotherapeutic sensitivity in bladder carcinoma was explored. By using immunohistochemistry, the expression of XIAP was detected in 47 bladder carcinomas and 5 normal bladder tissues. The XIAP gene was transfected into bladder cancer cell line T24 by liposome and the positive clone was screened by G418. Cellular XIAP mRNA level was detected by RT-PCR. Low-dose mitocycin C was administered to induce the apoptosis of T24 cells. The in vitro growth of bladder carcinoma cells was analyzed by MTT colorimetry, and the apoptosis rate was assayed by TUNEL methods. It was found XIAP was moderately expressed in bladder carcinomas with the the positive rate being 78.73% (37/47), but the positive rate was not correlated with carcinoma stages and grades (P<0.05). XIAP mRNA level in transfected T24 cells was significantly increased by 3.8 times as compared with that in the cells not transfected with XIAP. After treatment with low-dose mitomycin C (0.005 and 0.05 mg/mL), the growth rate in XIAP no-transfected control group was increased by (11.60±0.25)% and (16.51±0.87)% (P<0.05), and the apoptosis rate was decreased by (10.1±0.2)% and (11.9±0.2%) (P<0.05) respectively as compared with XIAP transfected group. It was concluded that XIAP was expressed in most of bladder carcimoma samples. Overexpression of XIAP in T24 could significantly reduce the MMC-induced apoptosis of bladder carcinoma, suggesting its effect on the chemothera- peutic sensitivity of T24 cells.展开更多
Objective:Using isobaric tags for relative and absolute quantitation(i TRAQ)technology to study differential protein expression in the retinal tissue of DR mouse models,providing proteomic evidence at the protein leve...Objective:Using isobaric tags for relative and absolute quantitation(i TRAQ)technology to study differential protein expression in the retinal tissue of DR mouse models,providing proteomic evidence at the protein level for the pathogenesis of DR.Methods:Firstly,establish diabetic mice and DR mice models,use i TRAQ technology to detect the retinal tissue samples of normal control group and DR model group mice,label the total retinal proteins of mice with i TRAQ reagent,and analyze them using mass spectrometry technology.Evaluate the differential proteins of the two groups using BioWorks TM 3.0 software,and conduct interaction feature analysis on the different proteins using the STRING website.Results:The i TRAQ technology detected a total of 406 different proteins between the diabetes and normal control groups,with 19 of them closely related to retinal cell apoptosis.Among them,significantly different proteins include acyl-CoA dehydrogenase short chain specific(ACADS),ataxin-10(ATXN10),BCL-2-associated X protein(BAX),caspase-3(CASP3),collagen type IIα1 chain(COL4A2),glycyl-tRNA synthetase(GARS),glial fibrillary acidic protein(GFAP),legumain(LGMN),mucin-4(MUC4),N-myc downstream-regulated gene 1 protein(NDRG1),with ratios to internal controls in the normal group of 1.67,2.06,1.76,2.16,1.53,1.87,1.24,1.61,0.42,0.56,respectively;and there is a potential functional association between GFAP,CASP3,and BAX proteins.Conclusion:In DR mice retinas,there are abnormal changes in the expression of a large number of apoptosis-related proteins.i TRAQ technology can effectively screen out key apoptosis proteins,among which GFAP,CASP3,and BAX may have adverse effects on the progression of DR by participating in the apoptosis process.The application of i TRAQ technology can provide new technical support for proteomic research on apoptosis proteins.展开更多
AIM: To study the effect of senescence marker protein 30(SMP30) on the proliferation and apoptosis of human lens epithelial cell(HLEC) SRA01/04.METHODS: SMP30 overexpression(OE) and knock down(KD) type cell ...AIM: To study the effect of senescence marker protein 30(SMP30) on the proliferation and apoptosis of human lens epithelial cell(HLEC) SRA01/04.METHODS: SMP30 overexpression(OE) and knock down(KD) type cell lines were cultivated by using two groups regucalcin(RGN; SMP30) lentiviral vectors(LVRGN, LV-RGN-RNAi) and the respective negative control virus infect SRA01/04 cells. Western blot and real-time quantitative polymerase chain reaction(q-PCR) analysis were used to determine RGN overexpression and knock down efficiency. We use cell counting kit-8(CCK8) assay to measure cell viability and 5-bromodeoxyuridine(Brd U) assay to test cell proliferation. Cell cycle was measured by PI FACS assay and cell apoptosis was tested by Annexin V-APC assay through flow cytometry. We use Western blot to measure the content of caspase-3 in SRA01/04.RESULTS: We used PCR and Western blot techniques to determine the successful transfection of SMP30 OE and KD SRA01/04 cell lines. By CCK8, Brdu and PI FACS cell cycle assay, it was found that the SMP30 OE group promoted cell proliferation(P〈0.05) compared with the control group, and the KD group inhibited cell proliferation(P〈0.05). The results of Annexin V-APC signal staining detection indicated that compared with respective control group, the cell apoptosis rate was higher in KD group(P〈0.05) but lower in OE group(P〈0.01). The expression of caspase-3 was down-regulated in OE group through Western blot assay and up-regulated in KD group compared with respective control group. CONCLUSION: Proliferation of SRA01/04 was promoted by SMP30 OE and apoptosis was suppressed. Increasing the expression of SMP30 may protect HLEC SRA01/04 against apoptosis in cataract.展开更多
Objective The cellular apoptosis susceptibility(CAS) protein plays a regulatory role in the induction of cell death in tumor cells. The objective of this study was to investigate the association of the expression of C...Objective The cellular apoptosis susceptibility(CAS) protein plays a regulatory role in the induction of cell death in tumor cells. The objective of this study was to investigate the association of the expression of CAS protein with HBV infection in the development of HCC. Methods The expression level of CAS was measured with immunohistochemistry. The occurrence of HBs Ag, HBe Ag and HBV DNA in HCC were concurrently examined with immunohistochemistry and in situ hybridization, respectively. Results The results showed that the CAS protein was detected in 86%(43/50), 70%(7/10), 15%(3/20) and none(0/20) of livers from patients with HCC, cholangiocarcinoma, cirrhosis and hepatitis, respectively. Furthermore, the level of CAS protein was higher in poorly differentiated tumors than moderately or well differentiated HCC. Interestingly, the CAS was stained significantly stronger in HBV-infected HCC than in non-HBV infected tissues(P < 0.01). Conclusions The expression of CAS is facilitated by HBV infection in HCC, suggesting that CAS might be a prognostic marker and a putative therapeutic target for HCC.展开更多
Septic encephalopathy is a frequent complication of sepsis,but there are few studies examining the role of micro RNAs(mi Rs) in its pathogenesis.In this study,a mi R-219 mimic was transfected into rat hippocampal ne...Septic encephalopathy is a frequent complication of sepsis,but there are few studies examining the role of micro RNAs(mi Rs) in its pathogenesis.In this study,a mi R-219 mimic was transfected into rat hippocampal neurons to model mi R-219 overexpression.A protective effect of mi R-219 was observed for glutamate-induced neurotoxicity of rat hippocampal neurons,and an underlying mechanism involving calmodulin-dependent protein kinase II γ(Ca MKIIγ) was demonstrated.mi R-219 and Ca MKIIγ m RNA expression induced by glutamate in hippocampal neurons was determined by quantitative real-time reverse transcription-polymerase chain reaction(q RT-PCR).After neurons were transfected with mi R-219 mimic,effects on cell viability and apoptosis were measured by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide(MTT) assay and flow cytometry.In addition,a luciferase reporter gene system was used to confirm Ca MKIIγ as a target gene of mi R-219.Western blot assay and rescue experiments were also utilized to detect Ca MKIIγ expression and further verify that mi R-219 in hippocampal neurons exerted its effect through regulation of Ca MKIIγ.MTT assay and q RT-PCR results revealed obvious decreases in cell viability and mi R-219 expression after glutamate stimulation,while Ca MKIIγ m RNA expression was increased.MTT,flow cytometry,and caspase-3 activity assays showed that mi R-219 overexpression could elevate glutamate-induced cell viability,and reduce cell apoptosis and caspase-3 activity.Moreover,luciferase Ca MKIIγ-reporter activity was remarkably decreased by co-transfection with mi R-219 mimic,and the results of a rescue experiment showed that Ca MKIIγ overexpression could reverse the biological effects of mi R-219.Collectively,these findings verify that mi R-219 expression was decreased in glutamate-induced neurons,Ca MKIIγ was a target gene of mi R-219,and mi R-219 alleviated glutamate-induced neuronal excitotoxicity by negatively controlling Ca MKIIγ expression.展开更多
Electroacupuncture has therapeutic effects on ischemic brain injury, but its mechanism is still poorly understood. In this study, mice were stimulated by electroacupuncture at the Baihui(GV20) acupoint for 30 minute...Electroacupuncture has therapeutic effects on ischemic brain injury, but its mechanism is still poorly understood. In this study, mice were stimulated by electroacupuncture at the Baihui(GV20) acupoint for 30 minutes at 1 m A and 2/15 Hz for 5 consecutive days. A cerebral ischemia model was established by ligating the bilateral common carotid artery for 15 minutes. At 72 hours after injury, neuronal injury in the mouse hippocampus had lessened, and the number of terminal deoxynucleotide transferase-mediated d UTP nick-end labeling-positive cells reduced after electroacupuncture treatment. Moreover, expression of adenosine monophosphate-activated protein kinase α(AMPKα) and phosphorylated AMPKα was up-regulated. Intraperitoneal injection of the AMPK antagonist, compound C, suppressed this phenomenon. Our findings suggest that electroacupuncture preconditioning alleviates ischemic brain injury via AMPK activation.展开更多
Background: Effective methods for managing patients with solitary pulmonary nodules(SPNs) depend critically on the predictive probability of malignancy.Methods: Between July 2009 and June 2011, data on gender, age...Background: Effective methods for managing patients with solitary pulmonary nodules(SPNs) depend critically on the predictive probability of malignancy.Methods: Between July 2009 and June 2011, data on gender, age, cancer history, tumor familial history, smoking status, tumor location, nodule size, spiculation, calcification, the tumor border, and the final pathological diagnosis were collected retrospectively from 154 surgical patients with an SPN measuring 3-30 mm. Each final diagnosis was compared with the probability calculated by three predicted models—the Mayo, VA, and Peking University(PU) models. The accuracy of each model was assessed using area under the receiver operating characteristics(ROC) and calibration curves.Results: The area under the ROC curve of the PU model [0.800; 95% confidence interval(CI): 0.708-0.891] was higher than that of the Mayo model(0.753; 95% CI: 0.650-0.857) or VA model(0.728; 95% CI: 0.623-0.833); however, this finding was not statistically significant. To varying degrees, calibration curves showed that all three models overestimated malignancy.Conclusions: The three predicted models have similar accuracy for prediction of SPN malignancy, although the accuracy is not sufficient. For Chinese patients, the PU model may has greater predictive power.Background: Here, we introduced our short experience on the application of a new CUSA Excel ultrasonic aspiration system, which was provided by Integra Lifesciences corporation, in skull base meningiomas resection.Methods: Ten patients with anterior, middle skull base and sphenoid ridge meningioma were operated using the CUSA Excel ultrasonic aspiration system at the Neurosurgery Department of Shanghai Huashan Hospital from August 2014 to October 2014. There were six male and four female patients, aged from 38 to 61 years old(the mean age was 48.5 years old). Five cases with tumor located at anterior skull base, three cases with tumor on middle skull base, and two cases with tumor on sphenoid ridge.Results: All the patents received total resection of meningiomas with the help of this new tool, and the critical brain vessels and nerves were preserved during operations. All the patients recovered well after operation.Conclusions: This new CUSA Excel ultrasonic aspiration system has the advantage of preserving vital brain arteries and cranial nerves during skull base meningioma resection, which is very important for skull base tumor operations. This key step would ensure a well prognosis for patients. We hope the neurosurgeons would benefit from this kind of technique.Background: The purposes of this study were to explore the effects of high mobility group protein box 1(HMGB1) gene on the growth, proliferation, apoptosis, invasion, and metastasis of glioma cells, with an attempt to provide potential therapeutic targets for the treatment of glioma. Methods: The expressions of HMGB1 in glioma cells(U251, U-87 MG and LN-18) and one control cell line(SVG p12) were detected by real time PCR and Western blotting, respectively. Then, the effects of HMGB1 on the biological behaviors of glioma cells were detected: the expression of HMGB1 in human glioma cell lines U251 and U-87 MG were suppressed using RNAi technique, then the influences of HMGB1 on the viability, cycle, apoptosis, and invasion abilities of U251 and U-87 MG cells were analyzed using in a Transwell invasion chamber. Also, the effects of HMGB1 on the expressions of cyclin D1, Bax, Bcl-2, and MMP 9 were detected. Results: As shown by real-time PCR and Western blotting, the expression of HMGB1 significantly increased in glioma cells(U251, U-87 MG, and LN-18) in comparison with the control cell line(SVG p12); the vitality, proliferation and invasive capabilities of U251 and U-87 MG cells in the HMGB1 siR NA-transfected group were significantly lower than those in the blank control group and negative control(NC) siR NA group(P〈0.05) but showed no significant difference between the blank control group and NC siR NA group. The percentage of apoptotic U251 and U-87 MG cells was significantly higher in the HMGB1 siR NA-transfected group than in the blank control group and NC siR NA group(P〈0.05) but was similar between the latter two groups. The HMGB1 siR NA-transfected group had significantly lower expression levels of Cyclin D1, Bcl-2, and MMP-9 protein in U251 and U-87 MG cells and significantly higher expression of Bax protein than in the blank control group and NC siR NA group(P〈0.05); the expression profiles of cyclin D1, Bax, Bcl-2, and MMP 9 showed no significant change in both blank control group and NC siR NA group. Conclusions: HMGB1 gene may promote the proliferation and migration of glioma cells and suppress its effects of apoptosis. Inhibition of the expression of HMGB1 gene can suppress the proliferation and migration of glioma cells and promote their apoptosis. Our observations provided a new target for intervention and treatment of glioma.展开更多
Objective: To study the expression and clinical value of apoptosis control gene bcl-2 and bax in breast cancer. Methods: Protein bax and bcl-2 in 41 breast cancers obtained from operations in our hospital in 1996 were...Objective: To study the expression and clinical value of apoptosis control gene bcl-2 and bax in breast cancer. Methods: Protein bax and bcl-2 in 41 breast cancers obtained from operations in our hospital in 1996 were detected using ABC immunohistochemical stain assay and compared with 10 cases with normal breast tissues. Results: The positive rate of bax in normal breast tissue was 90% and in breast cancer was 59%, with a significant statistical difference between them (P<0.05), but there was no statistical difference in bcl-2 protein expression. Among the 41 breast cancer, the group with lymph node metastasis (21 cases) had obviously low bax expression (43%) and high bcl-2 expression (76%), showing significant difference to the group without lymph node metastasis (P<0.05). Conclusion: The antiapoptosis function of bcl-2 was stronger than bax in breast cancer. Protein bax and bcl-2 assay may be useful in understanding the biological behaviors of breast cancer.展开更多
Polyethyleneimine-polyethylene glycol (PEI-PEG), a novel nanocarrier, has been used for trans- fection and gene therapy in a variety of cells. In our previous study, we successfully carried out PEI-PEG-mediated gene...Polyethyleneimine-polyethylene glycol (PEI-PEG), a novel nanocarrier, has been used for trans- fection and gene therapy in a variety of cells. In our previous study, we successfully carried out PEI-PEG-mediated gene transfer in spiral ganglion cells. It remains unclear whether PEI-PEG could be used for gene therapy with X-linked inhibitor of apoptosis protein (XIAP) in the inner ear. In the present study, we performed PEI-PEG-mediated XIAP gene transfection in the cochlea of Sprague-Dawley rats, via scala tympani fenestration, before daily cisplatin injections. Audito- ry brainstem reflex tests demonstrated the protective effects of XIAP gene therapy on auditory function. Immunohistochemical staining revealed XIAP protein expression in the cytoplasm of cells in the spiral ganglion, the organ of Corti and the stria vascularis. Reverse transcription-PCR detected high levels of XIAP mRNA expression in the cochlea. The present findings suggest that PEI-PEG nanocarrier-mediated XIAP gene transfection results in XIAP expression in the cochlea, prevents damage to cochlear spiral ganglion cells, and protects hearing.展开更多
Obstructive sleep apnea can worsen the prognosis of subarachnoid hemorrhage.Howeve r,the underlying mechanism remains unclear.In this study,we established a mouse model of subarachnoid hemorrhage using the endovascula...Obstructive sleep apnea can worsen the prognosis of subarachnoid hemorrhage.Howeve r,the underlying mechanism remains unclear.In this study,we established a mouse model of subarachnoid hemorrhage using the endovascular perforation method and exposed the mice to intermittent hypoxia for 8 hours daily for 2 consecutive days to simulate sleep apnea.We found that sleep apnea aggravated brain edema,increased hippocampal neuron apoptosis,and worsened neurological function in this mouse model of subarachnoid hemorrhage.Then,we established an in vitro HT-22 cell model of hemin-induced subarachnoid hemorrhage/intermittent hypoxia and found that the cells died,and lactate dehydrogenase release increased,after 48 hours.We further investigated the underlying mechanism and found that sleep apnea increased the expression of hippocampal neuroinflammatory factors interleukin-1β,interleukin-18,inte rleukin-6,nuclear factorκB,pyro ptosis-related protein caspase-1,pro-caspase-1,and NLRP3,promoted the prolife ration of astrocytes,and increased the expression of hypoxia-inducible factor 1αand apoptosis-associated speck-like protein containing a CARD,which are the key proteins in the hypoxia-inducible factor 1α/apoptosis-associated speck-like protein containing a CARD signaling pathway.We also found that knockdown of hypoxia-inducible factor 1αexpression in vitro greatly reduced the damage to HY22 cells.These findings suggest that sleep apnea aggravates early brain injury after subarachnoid hemorrhage by aggravating neuroinflammation and pyroptosis,at least in part through the hypoxia-inducible factor 1α/apoptosis-associated speck-like protein containing a CARD signaling pathway.展开更多
Bladder carcinoma is the most common tumor in the urinary system. In 1996, a sampleinvestigation showed that bladder carcinoma, in which more than 90% was mainly primary bladder transitional cell carcinoma (BTCC), w...Bladder carcinoma is the most common tumor in the urinary system. In 1996, a sampleinvestigation showed that bladder carcinoma, in which more than 90% was mainly primary bladder transitional cell carcinoma (BTCC), was one of the ten highest mortality malignant tumors in China. Bladder carcinoma represented 2% of all malignant tumors and has the fifth most common malignancy in men in Europe and North America.展开更多
Several studies have investigated the protective functions of brain-derived neurotrophic factor(BDNF) in retinitis pigmentosa. However, a BDNF-based therapy for retinitis pigmentosa is not yet available. To develop ...Several studies have investigated the protective functions of brain-derived neurotrophic factor(BDNF) in retinitis pigmentosa. However, a BDNF-based therapy for retinitis pigmentosa is not yet available. To develop an efficient treatment for fundus disease, an eukaryotic expression plasmid was generated and used to transfect human 293 T cells to assess the expression and bioactivity of BDNF on acute retinal pigment epithelial-19(ARPE-19) cells, a human retinal epithelial cell line. After 96 hours of co-culture in a Transwell chamber, ARPE-19 cells exposed to BDNF secreted by 293 T cells were more viable than ARPE-19 cells not exposed to secreted BDNF. Western blot assay showed that Bax levels were downregulated and that Bcl-2 levels were upregulated in human ARPE-19 cells exposed to BDNF. Furthermore, 293 T cells transfected with the BDNF gene steadily secreted the protein. The powerful anti-apoptotic function of this BDNF may be useful for the treatment of retinitis pigmentosa and other retinal degenerative diseases.展开更多
Micro RNA-124(mi R-124) is abundantly expressed in neurons in the mammalian central nervous system, and plays critical roles in the regulation of gene expression during embryonic neurogenesis and postnatal neural di...Micro RNA-124(mi R-124) is abundantly expressed in neurons in the mammalian central nervous system, and plays critical roles in the regulation of gene expression during embryonic neurogenesis and postnatal neural differentiation. However, the expression profile of mi R-124 after spinal cord injury and the underlying regulatory mechanisms are not well understood. In the present study, we examined the expression of mi R-124 in mouse brain and spinal cord after spinal cord injury using in situ hybridization. Furthermore, the expression of mi R-124 was examined with quantitative RT-PCR at 1, 3 and 7 days after spinal cord injury. The mi R-124 expression in neurons at the site of injury was evaluated by in situ hybridization combined with Neu N immunohistochemical staining. The mi R-124 was mainly expressed in neurons throughout the brain and spinal cord. The expression of mi R-124 in neurons significantly decreased within 7 days after spinal cord injury. Some of the neurons in the peri-lesion area were Neu N+/mi R-124-. Moreover, the neurons distal to the peri-lesion site were Neu N+/mi R-124+. These findings indicate that mi R-124 expression in neurons is reduced after spinal cord injury, and may reflect the severity of spinal cord injury.展开更多
Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder characterized by degeneration and loss of anterior horn cells in the spinal cord and brain stem nuclei, leading to progressive limb and ...Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder characterized by degeneration and loss of anterior horn cells in the spinal cord and brain stem nuclei, leading to progressive limb and trunk paralysis and muscular atrophy. Depending on the age of onset and maximum muscular function achieved, SMA is recognized as SMA1, SMA2, SMA3 or SMA4, and most patients have a deletion or truncation of the survival motor neuron 1 (SMN1) gene. In this report, we present a patient with a mild SMA phenotype, SMA3, and define his genetic abnormality. Tetra-primer amplification refractory mutation system PCR combined with restriction fragment length polymorphism analysis and array comparative genomic hybridization were used to determine the genetic variations in this patient. A 500 kb deletion in chromosome 5q13.2, including homozygous deletion of neuronal apoptosis inhibitory protein, and heterozygous deletion of occludin and B-double prime 1 was identified. This SMA region deletion did not involve SMN, indicating that SMN was likely to function normally. The phenotype was dependent of the large deletion and neuronal apoptosis inhibitory protein, occludin and B-double prime 1 may be candidate genes for SMA3.展开更多
Objectiv:To observe the effects ofRadix Salviae Miltiorrhizae (SM) on humanfibroblast in culture of kidney with lupus nephritis (LN ). Methods: Fibroblasts wereisolated from culture of kidney biopsy of LN patients, an...Objectiv:To observe the effects ofRadix Salviae Miltiorrhizae (SM) on humanfibroblast in culture of kidney with lupus nephritis (LN ). Methods: Fibroblasts wereisolated from culture of kidney biopsy of LN patients, and effect of SM on 3H-TdR incorporated rate of fibroblasts was observed. Theapoptosis and c-myc expression were detectedin the same time by flow cytometry.Results:SM could inhibit the proliferation of fibrolast,and promote the programmed cell deaththrough upregulate c-myc protein expression inhuman renal fibroblasts. Conclusions: Longterm administration of SM in large dosagecould be effective on interstial fibrosis of LN,so that to prevent or reduce the scar tissue for-mation and teatrd the occurrence of uremia.展开更多
Background:Circular RNAs(circRNAs)are considered to be important regulators in cancer biology.In this study,we focused on the effect of circRNA baculoviral inhibitor of apoptosis protein(IAP)repeat containing 6(circBI...Background:Circular RNAs(circRNAs)are considered to be important regulators in cancer biology.In this study,we focused on the effect of circRNA baculoviral inhibitor of apoptosis protein(IAP)repeat containing 6(circBIRC6)on non-small cell lung cancer(NSCLC)progression.Methods:The NSCLC and adjacent non-tumor tissues were collected at Shanghai Ninth People's Hospital.Quantitative real-time polymerase chain reaction was conducted for assessing the levels of circBIRC6,amyloid beta precursor protein binding protein 2(APPBP2)messenger RNA(mRNA),baculoviral IAP repeat containing 6 mRNA(BIRC6),and microRNA-217(miR-217).Western blot assay was adopted for measuring the protein levels of APPBP2,E-cadherin,N-cadherin,and vimentin.Colony formation assay,transwell assay,and flow cytometry analysis were utilized for evaluating cell colony formation,metastasis,and apoptosis.Dualluciferase reporter assay and RNA immunoprecipitation assay were carried out to determine the interaction between miR-217 and circBIRC6 and APPBP2 in NSCLC tissues.The murine xenograft model assay was used to investigate the function of circBIRC6 in tumor formation in vivo.Differences were analyzed via Student's t test or one-way analysis of variance.Pearson's correlation coefficient analysis was used to analyze linear correlation.Results:CircBIRC6 was overexpressed in NSCLC tissues and cells.Knockdown of circBIRC6 repressed the colony formation and metastasis and facilitated apoptosis of NSCLC cells in vitro and restrained tumorigenesis in vivo.Mechanically,circBIRC6 functioned as miR-217 sponge to promote APPBP2 expression in NSCLC cells.MiR-217 inhibition rescued circBIRC6 knockdown-mediated effects on NSCLC cell colony formation,metastasis,and apoptosis.Overexpression of miR-217 inhibited the malignant phenotypes of NSCLC cells,while the effects were abrogated by elevating APPBP2.Conclusion:CircBIRC6 aggravated NSCLC cell progression by elevating APPBP2 via sponging miR-217,which might provide a fresh perspective on NSCLC therapy.展开更多
文摘BACKGROUND The poor prognosis and rising incidence of esophageal cancer highlight the need for improved therapeutics that are essential prior to treatment.LCL161 is an SMAC(second mitochondrial activator of caspases)mimic and inhibitor of apoptosis protein(IAP)antagonist which exhibits anti-tumor effects and improves the chemical sensitivity of many cancers.AIM To ascertain the effects and mechanisms of the SMAC analog LCL161 on esophageal cancer cells.METHODS MTT assay and TUNEL assay were used to detect cell proliferation and apoptosis,respectively.Western blot analysis was used to study the molecular mechanisms of LCL161-induced death of ECA109 cells.RESULTS LCL161 decreased ECA109 cell proliferation in dose-and time-dependent manner and induced apoptosis of ECA109 cells in a dose-dependent manner.Also,LCL161 induced a significant decrease in the expression of the XIAP and significant increase in the expression of Caspase-3.In addition,Bax increased significantly with increasing concentrations of LCL161,and the relative expression of Bax was significantly different between groups.CONCLUSION These findings support the hypothesis that LCL161 can inhibit proliferation and induce apoptosis in esophageal cancer cells by regulating the expression of IAP family members,suggesting that it has potential to be an effective treatment for esophageal squamous cell carcinoma.
文摘Studies concerning correlations between pituitary adenomas and cell apoptosis have mainly focused on upstream apoptosis signaling, but seldom on downstream mediators. In the present study, second mitochondria-derived activator of caspases (Smac), X-linked inhibitor of apoptosis protein (XIAP), and caspase-3 protein were qualitatively analyzed using immunohistochemistry, and quantified by western blot. Smac, XIAP, and caspase-3 mRNA expressions were detected by reverse transcription-PCR. Results showed that XIAP protein and mRNA expressions were greater in the invasive pituitary adenoma group compared with the noninvasive pituitary adenoma group. However, Smac and caspase-3 protein and mRNA expressions were lower in the invasive pituitary adenoma group compared with the noninvasive pituitary adenoma group. In the invasive pituitary adenomas, Smac expression was positively correlated with caspase-3 protein and mRNA expression (Protein: r = 0.55, P 0.01; mRNA: r = 0.50, P 0.01). Smac and caspase-3 expressions were negatively correlated with XIAP protein and mRNA expression (Protein: r = -0.56, -0.64, P 0.01; mRNA: r = -0.69, -0.67, P 0.01). However, no significant differences in correlation among Smac, XIAP, and caspase-3 were detectable in noninvasive pituitary adenomas. These data indicated that high expression of XIAP and low expression of Smac and caspase-3 suppressed cell apoptosis and led to enhanced invasiveness of pituitary adenomas. Thus, Smac, XIAP, and caspase-3 may be useful markers in determining the invasive behavior of pituitary adenomas.
基金supported by the National Natural Science Foundation of China,Nos.31971277,31950410551Scientific Research Foundation for Returned Scholars+2 种基金Ministry of Education of ChinaPriority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)the Postgraduate Research&Practice Innovation Program of Jiangsu Province of China,No.KYCX 19-2050(all to DBY)。
文摘Wallerian degeneration is a complex biological process that occurs after nerve injury,and involves nerve degeneration and regeneration.Schwann cells play a crucial role in the cellular and molecular events of Wallerian degeneration of the peripheral nervous system.However,Wallerian degeneration regulating nerve injury and repair remains largely unknown,especially the early response.We have previously reported some key regulators of Wallerian degeneration after sciatic nerve injury.Baculoviral inhibitor of apoptosis protein repeat-containing protein 3(BIRC3)is an important factor that regulates apoptosis-inhibiting protein.In this study,we established rat models of right sciatic nerve injury.In vitro Schwann cell models were also established and subjected to gene transfection to inhibit and overexpress BIRC3.The data indicated that BIRC3 expression was significantly up-regulated after sciatic nerve injury.Both BIRC3 upregulation and downregulation affected the migration,proliferation and apoptosis of Schwan cells and affected the expression of related factors through activating c-fos and ERK signal pathway.Inhibition of BIRC3 delayed early Wallerian degeneration through inhibiting the apoptosis of Schwann cells after sciatic nerve injury.These findings suggest that BIRC3 plays an important role in peripheral nerve injury repair and regeneration.The study was approved by the Institutional Animal Care and Use Committee of Nantong University,China(approval No.2019-nsfc004)on March 1,2019.
基金supported by the National Natural Science Foundation of China (Grant No. 11071282)the Chinese Program for New Century Excellent Talents in University (Grant No. NCET-08-06867)+4 种基金the Natural Science Foundation of Hunan Province of China(Grant No. 10JJ7001)the Lotus Scholars Program of Hunan Province of Chinathe Aid Program for Science and Technology Innovative Research Team in Higher Educational Institutions of Hunan Province of Chinathe Australian Research Council (GrantNo. DP0559807)the Postgraduate Research and Innovation Project of Hunan Province of China (Grant No. CX2010B243)
文摘Apoptosis proteins play an important role in the development and homeostasis of an organism. The elucidation of the subcellular locations and functions of these proteins is helpful for understanding the mechanism of programmed cell death. In this paper, the recurrent quantification analysis, Hilbert-Huang transform methods, the maximum relevance and minimum redundancy method and support vector machine are used to predict the subcellular location of apoptosis proteins. The validation of the jackknife test suggests that the proposed method can improve the prediction accuracy of the subcellular location of apoptosis proteins and its application may be promising in other fields.
基金a grant from National Natu-ral Sciences Foundation of China (No. 30271301)
文摘The expression of X-linked inhibitor of apoptosis protein (XIAP) gene and its effect on chemotherapeutic sensitivity in bladder carcinoma was explored. By using immunohistochemistry, the expression of XIAP was detected in 47 bladder carcinomas and 5 normal bladder tissues. The XIAP gene was transfected into bladder cancer cell line T24 by liposome and the positive clone was screened by G418. Cellular XIAP mRNA level was detected by RT-PCR. Low-dose mitocycin C was administered to induce the apoptosis of T24 cells. The in vitro growth of bladder carcinoma cells was analyzed by MTT colorimetry, and the apoptosis rate was assayed by TUNEL methods. It was found XIAP was moderately expressed in bladder carcinomas with the the positive rate being 78.73% (37/47), but the positive rate was not correlated with carcinoma stages and grades (P<0.05). XIAP mRNA level in transfected T24 cells was significantly increased by 3.8 times as compared with that in the cells not transfected with XIAP. After treatment with low-dose mitomycin C (0.005 and 0.05 mg/mL), the growth rate in XIAP no-transfected control group was increased by (11.60±0.25)% and (16.51±0.87)% (P<0.05), and the apoptosis rate was decreased by (10.1±0.2)% and (11.9±0.2%) (P<0.05) respectively as compared with XIAP transfected group. It was concluded that XIAP was expressed in most of bladder carcimoma samples. Overexpression of XIAP in T24 could significantly reduce the MMC-induced apoptosis of bladder carcinoma, suggesting its effect on the chemothera- peutic sensitivity of T24 cells.
文摘Objective:Using isobaric tags for relative and absolute quantitation(i TRAQ)technology to study differential protein expression in the retinal tissue of DR mouse models,providing proteomic evidence at the protein level for the pathogenesis of DR.Methods:Firstly,establish diabetic mice and DR mice models,use i TRAQ technology to detect the retinal tissue samples of normal control group and DR model group mice,label the total retinal proteins of mice with i TRAQ reagent,and analyze them using mass spectrometry technology.Evaluate the differential proteins of the two groups using BioWorks TM 3.0 software,and conduct interaction feature analysis on the different proteins using the STRING website.Results:The i TRAQ technology detected a total of 406 different proteins between the diabetes and normal control groups,with 19 of them closely related to retinal cell apoptosis.Among them,significantly different proteins include acyl-CoA dehydrogenase short chain specific(ACADS),ataxin-10(ATXN10),BCL-2-associated X protein(BAX),caspase-3(CASP3),collagen type IIα1 chain(COL4A2),glycyl-tRNA synthetase(GARS),glial fibrillary acidic protein(GFAP),legumain(LGMN),mucin-4(MUC4),N-myc downstream-regulated gene 1 protein(NDRG1),with ratios to internal controls in the normal group of 1.67,2.06,1.76,2.16,1.53,1.87,1.24,1.61,0.42,0.56,respectively;and there is a potential functional association between GFAP,CASP3,and BAX proteins.Conclusion:In DR mice retinas,there are abnormal changes in the expression of a large number of apoptosis-related proteins.i TRAQ technology can effectively screen out key apoptosis proteins,among which GFAP,CASP3,and BAX may have adverse effects on the progression of DR by participating in the apoptosis process.The application of i TRAQ technology can provide new technical support for proteomic research on apoptosis proteins.
基金Supported by the National Natural Science Foundation of China(No.81360146)
文摘AIM: To study the effect of senescence marker protein 30(SMP30) on the proliferation and apoptosis of human lens epithelial cell(HLEC) SRA01/04.METHODS: SMP30 overexpression(OE) and knock down(KD) type cell lines were cultivated by using two groups regucalcin(RGN; SMP30) lentiviral vectors(LVRGN, LV-RGN-RNAi) and the respective negative control virus infect SRA01/04 cells. Western blot and real-time quantitative polymerase chain reaction(q-PCR) analysis were used to determine RGN overexpression and knock down efficiency. We use cell counting kit-8(CCK8) assay to measure cell viability and 5-bromodeoxyuridine(Brd U) assay to test cell proliferation. Cell cycle was measured by PI FACS assay and cell apoptosis was tested by Annexin V-APC assay through flow cytometry. We use Western blot to measure the content of caspase-3 in SRA01/04.RESULTS: We used PCR and Western blot techniques to determine the successful transfection of SMP30 OE and KD SRA01/04 cell lines. By CCK8, Brdu and PI FACS cell cycle assay, it was found that the SMP30 OE group promoted cell proliferation(P〈0.05) compared with the control group, and the KD group inhibited cell proliferation(P〈0.05). The results of Annexin V-APC signal staining detection indicated that compared with respective control group, the cell apoptosis rate was higher in KD group(P〈0.05) but lower in OE group(P〈0.01). The expression of caspase-3 was down-regulated in OE group through Western blot assay and up-regulated in KD group compared with respective control group. CONCLUSION: Proliferation of SRA01/04 was promoted by SMP30 OE and apoptosis was suppressed. Increasing the expression of SMP30 may protect HLEC SRA01/04 against apoptosis in cataract.
文摘Objective The cellular apoptosis susceptibility(CAS) protein plays a regulatory role in the induction of cell death in tumor cells. The objective of this study was to investigate the association of the expression of CAS protein with HBV infection in the development of HCC. Methods The expression level of CAS was measured with immunohistochemistry. The occurrence of HBs Ag, HBe Ag and HBV DNA in HCC were concurrently examined with immunohistochemistry and in situ hybridization, respectively. Results The results showed that the CAS protein was detected in 86%(43/50), 70%(7/10), 15%(3/20) and none(0/20) of livers from patients with HCC, cholangiocarcinoma, cirrhosis and hepatitis, respectively. Furthermore, the level of CAS protein was higher in poorly differentiated tumors than moderately or well differentiated HCC. Interestingly, the CAS was stained significantly stronger in HBV-infected HCC than in non-HBV infected tissues(P < 0.01). Conclusions The expression of CAS is facilitated by HBV infection in HCC, suggesting that CAS might be a prognostic marker and a putative therapeutic target for HCC.
基金supported by the National Natural Science Foundation of China,No.81101159the Natural Science Foundation of Jiangsu Province of China,No.BK20151268
文摘Septic encephalopathy is a frequent complication of sepsis,but there are few studies examining the role of micro RNAs(mi Rs) in its pathogenesis.In this study,a mi R-219 mimic was transfected into rat hippocampal neurons to model mi R-219 overexpression.A protective effect of mi R-219 was observed for glutamate-induced neurotoxicity of rat hippocampal neurons,and an underlying mechanism involving calmodulin-dependent protein kinase II γ(Ca MKIIγ) was demonstrated.mi R-219 and Ca MKIIγ m RNA expression induced by glutamate in hippocampal neurons was determined by quantitative real-time reverse transcription-polymerase chain reaction(q RT-PCR).After neurons were transfected with mi R-219 mimic,effects on cell viability and apoptosis were measured by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide(MTT) assay and flow cytometry.In addition,a luciferase reporter gene system was used to confirm Ca MKIIγ as a target gene of mi R-219.Western blot assay and rescue experiments were also utilized to detect Ca MKIIγ expression and further verify that mi R-219 in hippocampal neurons exerted its effect through regulation of Ca MKIIγ.MTT assay and q RT-PCR results revealed obvious decreases in cell viability and mi R-219 expression after glutamate stimulation,while Ca MKIIγ m RNA expression was increased.MTT,flow cytometry,and caspase-3 activity assays showed that mi R-219 overexpression could elevate glutamate-induced cell viability,and reduce cell apoptosis and caspase-3 activity.Moreover,luciferase Ca MKIIγ-reporter activity was remarkably decreased by co-transfection with mi R-219 mimic,and the results of a rescue experiment showed that Ca MKIIγ overexpression could reverse the biological effects of mi R-219.Collectively,these findings verify that mi R-219 expression was decreased in glutamate-induced neurons,Ca MKIIγ was a target gene of mi R-219,and mi R-219 alleviated glutamate-induced neuronal excitotoxicity by negatively controlling Ca MKIIγ expression.
基金supported by the National Natural Science Foundation of China,No.81273821
文摘Electroacupuncture has therapeutic effects on ischemic brain injury, but its mechanism is still poorly understood. In this study, mice were stimulated by electroacupuncture at the Baihui(GV20) acupoint for 30 minutes at 1 m A and 2/15 Hz for 5 consecutive days. A cerebral ischemia model was established by ligating the bilateral common carotid artery for 15 minutes. At 72 hours after injury, neuronal injury in the mouse hippocampus had lessened, and the number of terminal deoxynucleotide transferase-mediated d UTP nick-end labeling-positive cells reduced after electroacupuncture treatment. Moreover, expression of adenosine monophosphate-activated protein kinase α(AMPKα) and phosphorylated AMPKα was up-regulated. Intraperitoneal injection of the AMPK antagonist, compound C, suppressed this phenomenon. Our findings suggest that electroacupuncture preconditioning alleviates ischemic brain injury via AMPK activation.
文摘Background: Effective methods for managing patients with solitary pulmonary nodules(SPNs) depend critically on the predictive probability of malignancy.Methods: Between July 2009 and June 2011, data on gender, age, cancer history, tumor familial history, smoking status, tumor location, nodule size, spiculation, calcification, the tumor border, and the final pathological diagnosis were collected retrospectively from 154 surgical patients with an SPN measuring 3-30 mm. Each final diagnosis was compared with the probability calculated by three predicted models—the Mayo, VA, and Peking University(PU) models. The accuracy of each model was assessed using area under the receiver operating characteristics(ROC) and calibration curves.Results: The area under the ROC curve of the PU model [0.800; 95% confidence interval(CI): 0.708-0.891] was higher than that of the Mayo model(0.753; 95% CI: 0.650-0.857) or VA model(0.728; 95% CI: 0.623-0.833); however, this finding was not statistically significant. To varying degrees, calibration curves showed that all three models overestimated malignancy.Conclusions: The three predicted models have similar accuracy for prediction of SPN malignancy, although the accuracy is not sufficient. For Chinese patients, the PU model may has greater predictive power.Background: Here, we introduced our short experience on the application of a new CUSA Excel ultrasonic aspiration system, which was provided by Integra Lifesciences corporation, in skull base meningiomas resection.Methods: Ten patients with anterior, middle skull base and sphenoid ridge meningioma were operated using the CUSA Excel ultrasonic aspiration system at the Neurosurgery Department of Shanghai Huashan Hospital from August 2014 to October 2014. There were six male and four female patients, aged from 38 to 61 years old(the mean age was 48.5 years old). Five cases with tumor located at anterior skull base, three cases with tumor on middle skull base, and two cases with tumor on sphenoid ridge.Results: All the patents received total resection of meningiomas with the help of this new tool, and the critical brain vessels and nerves were preserved during operations. All the patients recovered well after operation.Conclusions: This new CUSA Excel ultrasonic aspiration system has the advantage of preserving vital brain arteries and cranial nerves during skull base meningioma resection, which is very important for skull base tumor operations. This key step would ensure a well prognosis for patients. We hope the neurosurgeons would benefit from this kind of technique.Background: The purposes of this study were to explore the effects of high mobility group protein box 1(HMGB1) gene on the growth, proliferation, apoptosis, invasion, and metastasis of glioma cells, with an attempt to provide potential therapeutic targets for the treatment of glioma. Methods: The expressions of HMGB1 in glioma cells(U251, U-87 MG and LN-18) and one control cell line(SVG p12) were detected by real time PCR and Western blotting, respectively. Then, the effects of HMGB1 on the biological behaviors of glioma cells were detected: the expression of HMGB1 in human glioma cell lines U251 and U-87 MG were suppressed using RNAi technique, then the influences of HMGB1 on the viability, cycle, apoptosis, and invasion abilities of U251 and U-87 MG cells were analyzed using in a Transwell invasion chamber. Also, the effects of HMGB1 on the expressions of cyclin D1, Bax, Bcl-2, and MMP 9 were detected. Results: As shown by real-time PCR and Western blotting, the expression of HMGB1 significantly increased in glioma cells(U251, U-87 MG, and LN-18) in comparison with the control cell line(SVG p12); the vitality, proliferation and invasive capabilities of U251 and U-87 MG cells in the HMGB1 siR NA-transfected group were significantly lower than those in the blank control group and negative control(NC) siR NA group(P〈0.05) but showed no significant difference between the blank control group and NC siR NA group. The percentage of apoptotic U251 and U-87 MG cells was significantly higher in the HMGB1 siR NA-transfected group than in the blank control group and NC siR NA group(P〈0.05) but was similar between the latter two groups. The HMGB1 siR NA-transfected group had significantly lower expression levels of Cyclin D1, Bcl-2, and MMP-9 protein in U251 and U-87 MG cells and significantly higher expression of Bax protein than in the blank control group and NC siR NA group(P〈0.05); the expression profiles of cyclin D1, Bax, Bcl-2, and MMP 9 showed no significant change in both blank control group and NC siR NA group. Conclusions: HMGB1 gene may promote the proliferation and migration of glioma cells and suppress its effects of apoptosis. Inhibition of the expression of HMGB1 gene can suppress the proliferation and migration of glioma cells and promote their apoptosis. Our observations provided a new target for intervention and treatment of glioma.
文摘Objective: To study the expression and clinical value of apoptosis control gene bcl-2 and bax in breast cancer. Methods: Protein bax and bcl-2 in 41 breast cancers obtained from operations in our hospital in 1996 were detected using ABC immunohistochemical stain assay and compared with 10 cases with normal breast tissues. Results: The positive rate of bax in normal breast tissue was 90% and in breast cancer was 59%, with a significant statistical difference between them (P<0.05), but there was no statistical difference in bcl-2 protein expression. Among the 41 breast cancer, the group with lymph node metastasis (21 cases) had obviously low bax expression (43%) and high bcl-2 expression (76%), showing significant difference to the group without lymph node metastasis (P<0.05). Conclusion: The antiapoptosis function of bcl-2 was stronger than bax in breast cancer. Protein bax and bcl-2 assay may be useful in understanding the biological behaviors of breast cancer.
基金supported by the Natural Science Foundation of Guangdong Province in China,No.S2011040003553
文摘Polyethyleneimine-polyethylene glycol (PEI-PEG), a novel nanocarrier, has been used for trans- fection and gene therapy in a variety of cells. In our previous study, we successfully carried out PEI-PEG-mediated gene transfer in spiral ganglion cells. It remains unclear whether PEI-PEG could be used for gene therapy with X-linked inhibitor of apoptosis protein (XIAP) in the inner ear. In the present study, we performed PEI-PEG-mediated XIAP gene transfection in the cochlea of Sprague-Dawley rats, via scala tympani fenestration, before daily cisplatin injections. Audito- ry brainstem reflex tests demonstrated the protective effects of XIAP gene therapy on auditory function. Immunohistochemical staining revealed XIAP protein expression in the cytoplasm of cells in the spiral ganglion, the organ of Corti and the stria vascularis. Reverse transcription-PCR detected high levels of XIAP mRNA expression in the cochlea. The present findings suggest that PEI-PEG nanocarrier-mediated XIAP gene transfection results in XIAP expression in the cochlea, prevents damage to cochlear spiral ganglion cells, and protects hearing.
基金the Natural Science Foundation of Jiangsu Province(Youth Program),No.BK20190129National Scientific Program of Jiangsu Colleges and Universities of China,No.19KJB320012(both to LY)。
文摘Obstructive sleep apnea can worsen the prognosis of subarachnoid hemorrhage.Howeve r,the underlying mechanism remains unclear.In this study,we established a mouse model of subarachnoid hemorrhage using the endovascular perforation method and exposed the mice to intermittent hypoxia for 8 hours daily for 2 consecutive days to simulate sleep apnea.We found that sleep apnea aggravated brain edema,increased hippocampal neuron apoptosis,and worsened neurological function in this mouse model of subarachnoid hemorrhage.Then,we established an in vitro HT-22 cell model of hemin-induced subarachnoid hemorrhage/intermittent hypoxia and found that the cells died,and lactate dehydrogenase release increased,after 48 hours.We further investigated the underlying mechanism and found that sleep apnea increased the expression of hippocampal neuroinflammatory factors interleukin-1β,interleukin-18,inte rleukin-6,nuclear factorκB,pyro ptosis-related protein caspase-1,pro-caspase-1,and NLRP3,promoted the prolife ration of astrocytes,and increased the expression of hypoxia-inducible factor 1αand apoptosis-associated speck-like protein containing a CARD,which are the key proteins in the hypoxia-inducible factor 1α/apoptosis-associated speck-like protein containing a CARD signaling pathway.We also found that knockdown of hypoxia-inducible factor 1αexpression in vitro greatly reduced the damage to HY22 cells.These findings suggest that sleep apnea aggravates early brain injury after subarachnoid hemorrhage by aggravating neuroinflammation and pyroptosis,at least in part through the hypoxia-inducible factor 1α/apoptosis-associated speck-like protein containing a CARD signaling pathway.
基金This work was supported by a grant from the National Nature ScienceFoundation of China (No.3000754)
文摘Bladder carcinoma is the most common tumor in the urinary system. In 1996, a sampleinvestigation showed that bladder carcinoma, in which more than 90% was mainly primary bladder transitional cell carcinoma (BTCC), was one of the ten highest mortality malignant tumors in China. Bladder carcinoma represented 2% of all malignant tumors and has the fifth most common malignancy in men in Europe and North America.
基金supported by the National Natural Science Foundation of China,No.81271046the Joint Program of Beijing Municipal Natural Science Foundation(category B)Beijing Educational Committee(key project),No.KZ201510025025
文摘Several studies have investigated the protective functions of brain-derived neurotrophic factor(BDNF) in retinitis pigmentosa. However, a BDNF-based therapy for retinitis pigmentosa is not yet available. To develop an efficient treatment for fundus disease, an eukaryotic expression plasmid was generated and used to transfect human 293 T cells to assess the expression and bioactivity of BDNF on acute retinal pigment epithelial-19(ARPE-19) cells, a human retinal epithelial cell line. After 96 hours of co-culture in a Transwell chamber, ARPE-19 cells exposed to BDNF secreted by 293 T cells were more viable than ARPE-19 cells not exposed to secreted BDNF. Western blot assay showed that Bax levels were downregulated and that Bcl-2 levels were upregulated in human ARPE-19 cells exposed to BDNF. Furthermore, 293 T cells transfected with the BDNF gene steadily secreted the protein. The powerful anti-apoptotic function of this BDNF may be useful for the treatment of retinitis pigmentosa and other retinal degenerative diseases.
基金supported by the National Natural Science Foundation of China,No.81371364
文摘Micro RNA-124(mi R-124) is abundantly expressed in neurons in the mammalian central nervous system, and plays critical roles in the regulation of gene expression during embryonic neurogenesis and postnatal neural differentiation. However, the expression profile of mi R-124 after spinal cord injury and the underlying regulatory mechanisms are not well understood. In the present study, we examined the expression of mi R-124 in mouse brain and spinal cord after spinal cord injury using in situ hybridization. Furthermore, the expression of mi R-124 was examined with quantitative RT-PCR at 1, 3 and 7 days after spinal cord injury. The mi R-124 expression in neurons at the site of injury was evaluated by in situ hybridization combined with Neu N immunohistochemical staining. The mi R-124 was mainly expressed in neurons throughout the brain and spinal cord. The expression of mi R-124 in neurons significantly decreased within 7 days after spinal cord injury. Some of the neurons in the peri-lesion area were Neu N+/mi R-124-. Moreover, the neurons distal to the peri-lesion site were Neu N+/mi R-124+. These findings indicate that mi R-124 expression in neurons is reduced after spinal cord injury, and may reflect the severity of spinal cord injury.
基金the Foundation of Science and Technology Department of Zhejiang Province,China,No. 2007C33049Fund for Fostering Talents in Basic Science of the National Natural Science Foundation of China,No. J0710043
文摘Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder characterized by degeneration and loss of anterior horn cells in the spinal cord and brain stem nuclei, leading to progressive limb and trunk paralysis and muscular atrophy. Depending on the age of onset and maximum muscular function achieved, SMA is recognized as SMA1, SMA2, SMA3 or SMA4, and most patients have a deletion or truncation of the survival motor neuron 1 (SMN1) gene. In this report, we present a patient with a mild SMA phenotype, SMA3, and define his genetic abnormality. Tetra-primer amplification refractory mutation system PCR combined with restriction fragment length polymorphism analysis and array comparative genomic hybridization were used to determine the genetic variations in this patient. A 500 kb deletion in chromosome 5q13.2, including homozygous deletion of neuronal apoptosis inhibitory protein, and heterozygous deletion of occludin and B-double prime 1 was identified. This SMA region deletion did not involve SMN, indicating that SMN was likely to function normally. The phenotype was dependent of the large deletion and neuronal apoptosis inhibitory protein, occludin and B-double prime 1 may be candidate genes for SMA3.
文摘Objectiv:To observe the effects ofRadix Salviae Miltiorrhizae (SM) on humanfibroblast in culture of kidney with lupus nephritis (LN ). Methods: Fibroblasts wereisolated from culture of kidney biopsy of LN patients, and effect of SM on 3H-TdR incorporated rate of fibroblasts was observed. Theapoptosis and c-myc expression were detectedin the same time by flow cytometry.Results:SM could inhibit the proliferation of fibrolast,and promote the programmed cell deaththrough upregulate c-myc protein expression inhuman renal fibroblasts. Conclusions: Longterm administration of SM in large dosagecould be effective on interstial fibrosis of LN,so that to prevent or reduce the scar tissue for-mation and teatrd the occurrence of uremia.
文摘Background:Circular RNAs(circRNAs)are considered to be important regulators in cancer biology.In this study,we focused on the effect of circRNA baculoviral inhibitor of apoptosis protein(IAP)repeat containing 6(circBIRC6)on non-small cell lung cancer(NSCLC)progression.Methods:The NSCLC and adjacent non-tumor tissues were collected at Shanghai Ninth People's Hospital.Quantitative real-time polymerase chain reaction was conducted for assessing the levels of circBIRC6,amyloid beta precursor protein binding protein 2(APPBP2)messenger RNA(mRNA),baculoviral IAP repeat containing 6 mRNA(BIRC6),and microRNA-217(miR-217).Western blot assay was adopted for measuring the protein levels of APPBP2,E-cadherin,N-cadherin,and vimentin.Colony formation assay,transwell assay,and flow cytometry analysis were utilized for evaluating cell colony formation,metastasis,and apoptosis.Dualluciferase reporter assay and RNA immunoprecipitation assay were carried out to determine the interaction between miR-217 and circBIRC6 and APPBP2 in NSCLC tissues.The murine xenograft model assay was used to investigate the function of circBIRC6 in tumor formation in vivo.Differences were analyzed via Student's t test or one-way analysis of variance.Pearson's correlation coefficient analysis was used to analyze linear correlation.Results:CircBIRC6 was overexpressed in NSCLC tissues and cells.Knockdown of circBIRC6 repressed the colony formation and metastasis and facilitated apoptosis of NSCLC cells in vitro and restrained tumorigenesis in vivo.Mechanically,circBIRC6 functioned as miR-217 sponge to promote APPBP2 expression in NSCLC cells.MiR-217 inhibition rescued circBIRC6 knockdown-mediated effects on NSCLC cell colony formation,metastasis,and apoptosis.Overexpression of miR-217 inhibited the malignant phenotypes of NSCLC cells,while the effects were abrogated by elevating APPBP2.Conclusion:CircBIRC6 aggravated NSCLC cell progression by elevating APPBP2 via sponging miR-217,which might provide a fresh perspective on NSCLC therapy.