CNKSR2 interactome analysis indicates its association with the centrosome/microtubule system

The protein connector enhancer of kinase suppressor of Ras 2(CNKSR2),present in both the postsynaptic density and cytoplasm of neurons,is a scaffolding protein with several protein-binding domains.Variants of the CNKSR2 gene have been implicated in neurodevelopmental disorders,particularly intellectual disability,although the precise mechanism involved has not yet been fully understood.Research has demonstrated that CNKSR2 plays a role in facilitating the localization of postsynaptic density protein complexes to the membrane,thereby influencing synaptic signaling and the morphogenesis of dendritic spines.However,the function of CNKSR2 in the cytoplasm remains to be elucidated.In this study,we used immunoprecipitation and high-resolution liquid chromatography-mass spectrometry to identify the interactors of CNKSR2.Through a combination of bioinformatic analysis and cytological experiments,we found that the CNKSR2 interactors were significantly enriched in the proteome of the centrosome.We also showed that CNKSR2 interacted with the microtubule protein DYNC1H1 and with the centrosome marker CEP290.Subsequent colocalization analysis confirmed the centrosomal localization of CNKSR2.When we downregulated CNKSR2 expression in mouse neuroblastoma cells(Neuro 2A),we observed significant changes in the expression of numerous centrosomal genes.This manipulation also affected centrosome-related functions,including cell size and shape,cell proliferation,and motility.Furthermore,we found that CNKSR2 interactors were highly enriched in de novo variants associated with intellectual disability and autism spectrum disorder.Our findings establish a connection between CNKSR2 and the centrosome,and offer new insights into the underlying mechanisms of neurodevelopmental disorders.

Physical Activity,Sedentary Behavior,and the Risk of Cardiovascular Disease in Type 2 Diabetes Mellitus Patients:The MIDiab Study

The aim of this study was to explore the associations of moderate-to-vigorous-intensity physical activity(MVPA)time and sedentary(SED)time with a history of cardiovascular disease(CVD)and multifactorial(i.e.,blood pressure(BP),body mass index(BMI),low-density lipoprotein cholesterol(LDL-C),and glycated hemoglobin A1c(HbA1c))control status among type 2 diabetes mellitus(T2DM)patients in China.A cross-sectional analysis of 9152 people with type 2 diabetes from the Multifactorial Intervention on Type 2 Diabetes(MIDiab)study was performed.Patients were grouped according to their self-reported MVPA time(low,<150 min·week−1;moderate,150 to<450 min·week−1;high,≥450 min·week−1)and SED time(low,<4 h·d–1;moderate,4 to<8 h·d–1;high,≥8 h·d–1).Participants who self-reported a history of CVD were identified as having a CVD risk.Odds ratios(ORs)and 95%confidence intervals(CIs)of CVD risk and multifactorial control status associated with MVPA time and SED time were estimated using mixed-effect logistic regression models,adjusting for China’s geographical region characteristics.The participants had a mean±standard deviation(SD)age of(60.87±8.44)years,44.5%were women,and 25.1%had CVD.After adjustment for potential confounding factors,an inverse association between high MVPA time and CVD risk that was independent of SED time was found,whereas this association was not observed in the moderate-MVPA group.A higher MVPA time was more likely to have a positive effect on the control of BMI.Compared with the reference group(i.e.,those with MVPA time≥450 min·week−1 and SED time<4 h·d–1),CVD risk was higher in the low-MVPA group:The OR associated with an SED time<4 h·d–1 was 1.270(95%CI,1.040–1.553)and that associated with an SED time≥8 h·d–1 was 1.499(95%CI,1.149–1.955).We found that a high MVPA time(i.e.,≥450 min·week−1)was associated with lower odds of CVD risk regardless of SED time among patients with T2DM.

Integrated analysis of single-cell and bulk RNA-seq establishes a novel signature for prediction in gastric cancer

BACKGROUND Single-cell sequencing technology provides the capability to analyze changes in specific cell types during the progression of disease.However,previous single-cell sequencing studies on gastric cancer(GC)have largely focused on immune cells and stromal cells,and further elucidation is required regarding the alterations that occur in gastric epithelial cells during the development of GC.AIM To create a GC prediction model based on single-cell and bulk RNA sequencing(bulk RNA-seq)data.METHODS In this study,we conducted a comprehensive analysis by integrating three singlecell RNA sequencing(scRNA-seq)datasets and ten bulk RNA-seq datasets.Our analysis mainly focused on determining cell proportions and identifying differentially expressed genes(DEGs).Specifically,we performed differential expression analysis among epithelial cells in GC tissues and normal gastric tissues(NAGs)and utilized both single-cell and bulk RNA-seq data to establish a prediction model for GC.We further validated the accuracy of the GC prediction model in bulk RNA-seq data.We also used Kaplan–Meier plots to verify the correlation between genes in the prediction model and the prognosis of GC.RESULTS By analyzing scRNA-seq data from a total of 70707 cells from GC tissue,NAG,and chronic gastric tissue,10 cell types were identified,and DEGs in GC and normal epithelial cells were screened.After determining the DEGs in GC and normal gastric samples identified by bulk RNA-seq data,a GC predictive classifier was constructed using the Least absolute shrinkage and selection operator(LASSO)and random forest methods.The LASSO classifier showed good performance in both validation and model verification using The Cancer Genome Atlas and Genotype-Tissue Expression(GTEx)datasets[area under the curve(AUC)_min=0.988,AUC_1se=0.994],and the random forest model also achieved good results with the validation set(AUC=0.92).Genes TIMP1,PLOD3,CKS2,TYMP,TNFRSF10B,CPNE1,GDF15,BCAP31,and CLDN7 were identified to have high importance values in multiple GC predictive models,and KM-PLOTTER analysis showed their relevance to GC prognosis,suggesting their potential for use in GC diagnosis and treatment.CONCLUSION A predictive classifier was established based on the analysis of RNA-seq data,and the genes in it are expected to serve as auxiliary markers in the clinical diagnosis of GC.

Practical Progress of SBAR Communication Model-Related Teaching in Clinical Teaching

This paper elaborates the core and essence of situation,background,assessment,recommendation(SBAR)communication model,clarify the application status of SBAR communication model in the clinical teaching of nursing students,and describe its effectiveness and limitations,so as to provide reference for improving clinical nursing teaching ability.

The Latest Research Progress in the Application of MEWS Scoring System in Clinical Nursing

This paper summarizes the background of the formation of the Modified Early Warning System(MEWS)evaluation system,its current status of clinical teaching applications in different fields of hospitals,and its significance on the medical and nursing career,aiming to provide specific theoretical basis for medical staff and lay a foundation for continuing to carry out related work on MEWS.

The Clinical Pathologic Analysis of Radiotherapy for Cutaneous B-cell Lymphoma

OBJECTIVE To report results of radiation therapy treatment of 30 B-cell lymphoma patients with an initial cutaneous presentation according to the new classification by the WHO/EORTC. METHODS Thirty patients with cutaneous B-cell lymphoma （CBCL） were treated by cutaneous irradiation based on the number and location of the lesions and the stage of their tumor. Treatment was conducted using a Satume Clinac. RESULTS A complete response （CR） from the treatment for our series was 86%. The length of complete remission ranged from 4 to 301 months. Three patients （11%） developed a partial response （PR）. One patient was progressive. Disease-free survival（DFS） at 10 years was 87%. Three patiens died [One PCMZL two PCLBCL leg type （29%）]. Radiotherapy was generally well tolerated. CONCLUSION According to the WHO/EORTC classification, the survivor results were good for PCMZL and PCFCL. The PCLBCL leg type had a poor prognosis. Localized field irradiation is an effective treatment for some localized forms of primary cutaneous B-cell lymphoma, and this mode of therapy can produce prolonged remissions.The patients with wide-spread skin involvement are usually candidates for extended field irradiation and/or chemotherapy. For advanced stages of cutaneous B-cell lymphoma, where chemotherapy is the treatment of choice, a degree of palliation can be achieved using local field irradation.

Aberrant TGF-β1 signaling contributes to the development of primary biliary cirrhosis in murine model

AIM:To investigate whether transforming growth factor-β1(TGF-β1)signaling pathway is involved in the pathogenesis of primary biliary cirrhosis(PBC).METHODS:A murine model of PBC was developed by injection of polyinosinic polycytidylic acids(polyⅠ:C)in C57BL/6 mice,and the liver expressions of TGFβ1,TGF-βreceptorⅠ(TβRⅠ),TGF-βreceptorⅡ(TβRⅡ),p-Smad2/3,monoclonalα-smooth muscle actin antibody(α-SMA)andα1(Ⅰ)collagen in the mouse model and control mice were evaluated by immunohistochemistry,immunoblotting and real-time polymerase chain reaction(RT-PCR).Lymphocyte subsets in liver were analyzed using flow cytometry.RESULTS:The mouse model had several key phenotypic features of human PBC,including elevated levels of alkaline phosphatase,antimitochondrial antibodies,portal bile ducts inflammation,and progressive collagen deposition.Compared with control mice,protein and mRNA levels of TGFβ1,TβRⅠ,TβRⅡ,p-Smad2/3,α-SMA andα1(Ⅰ)collagen in liver(1.7±0.4 vs 8.9±1.8,0.8±0.2 vs 5.1±1.5,0.6±0.01 vs5.1±0.1,0.6±0.3 vs 2.0±0.3,0.9±0.4 vs 3.4±0.6,0.8±0.4 vs 1.7±0.3,1.1±1.2 vs 11.8±0.6,P<0.05),and the total number and percentage of CD4+CD25+FOXP3+and CD8+lymphocytes(0.01±0.001vs 0.004±0.00,0.12±0.04 vs 0.52±0.23,P<0.01)were higher in the mouse model.CONCLUSION:TGFβ1 might play a dual role in the development of PBC:it suppresses inflammatory response but operates to enhance fibrogenesis.The aberrant activity of TGF-β1 signaling contributes to the development of PBC.

Picroside Ⅱ down-regulates matrix metalloproteinase-9 expression following cerebral ischemia/reperfusion injury in rats

Studies have shown that Picroside Ⅱ attenuates inflammatory reactions following brain ischemia through the inhibition of the TLR-4-NF-KB signal transduction pathway, and ameliorates cerebral edema through the reduction of aquaporin-4 expression. Matrix metalloproteinase-9 （MMP-9）, located downstream of the TLR-4-NF-KB signal transduction pathway, can degrade the neurovascular matrix, damage the blood-brain barrier to induce cerebral edema, and directly result in neuronal apoptosis and brain injury, Therefore, the present study further observed MMP-9 expression in the brain tissues of rats with cerebral ischemia/reperfusion injury following Picroside Ⅱ treatment. Results demonstrated that Picroside Ⅱ significantly reduced MMP-9 expression in ischemic brain tissues, as well as neuronal apoptosis and brain infarct volume, suggesting Picroside Ⅱ exhibits neuroprotection by down-regulating MMP-9 expression and inhibiting cell apoptosis.

Correlation of Epstein-Barr virus and its encoded proteins with Helicobacter pylori and expression of c-met and c-myc in gastric carcinoma

AIM： To investigate the interrelationship of Epstein-Barr virus （EBV） and EBV- encoded proteins with Helicobacter pylori （H pylor/~ infection and the expression of c-met and c-myc oncogene proteins in gastric carcinoma, and to explore their role in gastric carcinogenesis. METHODS： One hundred and eighty-five gastric carcinoma tissues were detected by polymerase chain reaction （PCR）-Southern blot for EBV genome and in situ hybridization （ISH） for EBV-encoded small RNA 1 （EBER1）. Gastric carcinoma with positive EBER1 signals was confirmed EBV-associated gastric carcinoma （EBVaGC）. The status of Hpylori infection in 185 gastric carcinomas was assessed by rapid urease test and PCR. The samples with positive PCR and urease test were defined as H pylorl infection. The expression of c-met and c-myc oncogene proteins in tissues of EBVaGC and matched EBV-negative gastric carcinoma （EBVnGC） were examined by immunohistochemistry. RT-PCR and Southern hybridization were used to detect the expression of nuclear antigens （EBNAs） 1 and 2, latent membrane protein （LMP） 1, early genes BARF1 and BHRF1 in EBVaGC cases. RESULTS： The positive rate of H pylori and EBV in 185 gastric carcinomas was 59.45% （110/185） and 7.03% （13/185） respectively. No difference was found in sex, age, pathological differentiation, clinical stages and lymph node metastasis between H pylori-positive and H pylori-negative gastric carcinomas. However, the positive rate of H pylori infection in the antrum gastric carcinomas was higher than that of cardia and body gastric carcinomas. In our series, age, pathological differentiation, clinical stages, lymph node metastasis and location of cancer were not different between EBVnGC and EBVaGC, while the positive rate of EBV in male patients was significantly higher than that of female patients. The positivity of Hpylori in EBV-associated and EBV-negative gastric carcinomas was 46.15% （6/13） and 81.40%（104/172） respectively. There was no significant correlation between EBV and H pylori infection. The c-met overexpression was significantly higher in the EBVaGC group than in the EBVnGC group. However, c-met and c-myc expression did not show significant difference between the two groups. Transcripts of EBNA1 were detected in all 13 EBVaGCs, while both EBNA2 and LMP1 mRNA were not detected. Six of the 13 cases exhibited BARF1 transcripts and 2 exhibited BHRF1 transcripts. CONCLUSION： The positivity of H pylori in EBVnGCs is higher than that of EBVaGCs, but no significant correlation is found between EBV infection and H pylori infection. H pylori-positive gastric carcinoma is predominant in antrum location, while EBVaGC has a tendency of predominance in cardia/body location. EBV infection is associated with c-met abnormal expression but not with c-myc protein in EBVaGC. c-met overexpression is not induced by LMP1. BARF1 and BHRF1 may play important roles in the tumorigenesis of EBVaGC through different pathways.

CXCR4/SDF-1 axis is involved in lymph node metastasis of gastric carcinoma

AIM:To investigate the role of CXC chemokine receptor-4 (CXCR4) and stromal cell-derived factor-1 (SDF-1) in lymph node metastasis of gastric carcinoma.METHODS:In 40 cases of gastric cancer,expression of CXCR4 mRNA in cancer and normal mucous membrane and SDF-1 mRNA in lymph nodes around the stomach was detected using quantitative polymerase chain reaction (PCR) (TaqMan) and immunohistochemistric assay.SGC-7901 and MGC80-3 cancer cells were used to investigate the effect of SDF-1 on cell proliferation and migration.RESULTS:Quantitative reverse transcription PCR and immunohistochemistry revealed that the expression level of CXCR4 in gastric cancer was significantly higher than that in normal mucous membrane (1.6244 ± 1.3801 vs 1.0715 ± 0.5243,P < 0.05).The expression level of CXCR4 mRNA in gastric cancer with lymph node metastasis was also significantly higher than that without lymph node metastasis (0.823 ± 0.551 vs 0.392 ± 0.338,P < 0.05).CXCR4 expression was significantly related to poorly differentiated,high tumor stage and lymph node metastasis.Significant differences in the expression level of SDF-1 mRNA were found between lymph nodes in metastatic gastric cancer and normal nodes (0.5432 ± 0.4907 vs 0.2640 ± 0.2601,P < 0.05).The positive expression of SDF-1 mRNA in lymph nodes of metastatic gastric cancer was consistent with the positive expression of CXCR4 mRNA in gastric cancer (r=0.776,P < 0.01).Additionally,human gastric cancer cell lines expressed CXCR4 and showed vigorous proliferation and migratory responses to SDF-1.AMD3100 (a specific CXCR4 antagonist) was also found to effectively reduce the migration of gastric cancer cells.CONCLUSION:The CXCR4/SDF-1 axis is involved in the lymph node metastasis of gastric cancer.CXCR4 is considered as a potential therapeutic target in the treatment of gastric cancer.

Gastric outlet obstruction caused by heterotopic pancreas:A case report and a quick review

A 46-year-old Chinese woman presented with nausea, recurrent vomiting, and abdominal pain. Gastroduodenal endoscopic examination revealed an oval-shaped submucosal tumor at the prepyloric area on the posterior wall of the stomach. A degenerated gastrointestinal stromal tumor was suspected. Distal gastrectomy was performed and a histological diagnosis of heterotopic pancreas （HPs） was confirmed. The patient had an uneventful postoperative course and was discharged 7 d a^er operation. The patient remains healthy and symptom-free in the follow-up of 6 mo. This is a report of a case of gastric outlet obstruction resulting from pancreatic heterotopia in the gastric antrum in an adult woman.

Anti-oxidant effect of picroside II in a rat model of cerebral ischemia/reperfusion injury

Picroside II,the major active component of picroside,has been shown to induce PC12 cell axonal growth and relieve free radical damage.In vivo experiments have demonstrated that picroside II can improve neurological function in rats with cerebral ischemia/reperfusion injuries.In the present in vivo study,enzyme-linked immunosorbent assay and immunohistochemistry revealed that picroside II increased superoxide dismutase content and reduced inducible nitric oxide synthase content in the ischemic hemisphere.The effects of picroside II were similar to those of salvianic acid A sodium,an active control drug.These results indicate that picroside II exerts a neuroprotective effect,possibly by downregulating inducible nitric oxide synthase expression,increasing superoxide dismutase activity,and inhibiting neuronal apoptosis.

Contralateral needling at unblocked collaterals for hemiplegia following acute ischemic stroke

Hemiplegia caused by stroke indicates dysfunction of the network between the brain and limbs, namely collateral shock in the brain. Contralateral needling is the insertion of needles into acupoints on the relative healthy side of the body to treat diseases such as apoplexy. However, there is little well-designed and controlled clinical evidence for this practice. This study investigated whether contralateral needling could treat hemiplegia after acute ischemic stroke in 106 randomly selected patients with acute ischemic stroke. These patients were randomly assigned to three groups： 45 in the contralateral needling group, receiving acupuncture on the unaffected limbs; 45 in the conven- tional acupuncture group, receiving acupuncture on the hemiplegic limbs; and 16 in the control group, receiving routine treatments without acupuncture. Acupuncture at acupoints Chize （LU5） in the upper limb and Jianliao （TEl4） in the lower limb was performed for 45 minutes daily for 30 consecutive days. The therapeutic effective rate, Neurological Deficit Score, Modified Barthel Index and FugI-Meyer Assessment were evaluated. The therapeutic effective rate of contralateral nee- dling was higher than that of conventional acupuncture （46.67% vs. 31.11%, P 〈 0.05）. The neuro- logical deficit score of contralateral needling was significantly decreased compared with conven- tional acupuncture （P 〈 0.01）. The Modified Barthel Index and FugI-Meyer Assessment score of contralateral needling increased more significantly than those of conventional acupuncture （both P 〈 0.01）. The present findings suggest that contralateral needling unblocks collaterals and might be more effective than conventional acupuncture in the treatment of hemiplegia following acute ischemic stroke.

Effect of PDGF-Rb antagonist imatinib on endometrial injury repairing in mouse model

Objective: To study the effects of PDGF-Rb antagonists imatinib on endometrial injury repairing in the mouse model. Methods: The cultured MSCs cells from male mice were marked with Brd U in vitro, and then transplanted to the female mice which suffered from radiation injury through tail vein, PDGF-Rb antagonists imatinib was injected through abdominal cavity. Four groups were arranged, which were radiation transplantation group, normal control group, imatinib intervention group and radiation control group. Brd U incorporation, SRY expression and MVD status were detected in uterus of mice. Results: SRY gene was negative expressed in normal control group and radiation control group. SRY gene presented positive in radiation transplantation group and imatinib intervention group; Brd U incorporation showed negative in radiation control group and normal control group which died in the early stage in mice; the incorporation of Brd U was higher in radiation transplantation group compared with imatinib intervention group; CD34 was positive on the uterus of all the four groups,which showed highest in radiation control group and lowest in radiation control group; The MVD in imatinib intervention group was lower than radiation control group; the difference of MVD was significantly compared with normal control group(P<0.05). Conclusions: PDGF-Rb antagonists imatinib could inhibit the repairing function of MSCs in the endometrial lesions in mice.

CD133^+ gallbladder carcinoma cells exhibit self-renewal ability and tumorigenicity

AIM： To identify cancer stern cells （CSCs） in human gallbladder carcinomas （GBCs）. METHODS： Primary GBC cells were cultured under serum-free conditions to produce floating spheres. The stem-cell properties of the sphere-forming cells, including self-renewal, differentiation potential, chemoresistance and tumorigenicity, were determined in vitro or in vivo. Cell surface expression of CD133 was investigated in primary tumors and in spheroid cells using flow cytometry. The sphere-colony-formation ability and tumorigenicity of CD133＋ cells were assayed.floating spheroids were generated from primary GBC cells, and these sphere-forming cells could generate new progeny spheroids in serum-free media. Spheroid cells were differentiated under serum-containing conditions with downregulation of the stem cell markers Oct-4, Nanog, and nestin （P 〈 0.05）. The differentiated cells showed lower spheroid-colony-formation ability than the original spheroid cells （P 〈 0.05）. Spheroid ceils were more resistant to chemotherapeutic reagents than the congenetic adherent cells （P 〈 0.05）. Flow cytometry showed enriched CD133＋ population in sphereforming cells （P 〈 0.05）. CD133＋ cells possessed high colony-formation ability than the CD133 population （P 〈 0.01）. CD133＋ cells injected into nude mice revealed higher tumorigenicity than their antigen-negative counterparts （P 〈 0.05）. CONCLUSION： CD133 may be a cell surface marker for CSCs in GBC.

Effects of acupuncturing Tsusanli (S_T36) on expression of nitric oxide synthase in hypothalamus and adrenal gland in rats with cold stress ulcer

AIM： To study the protective effect of acupuncturing Tsusanli （ST36） on cold stress ulcer, and the expression of nitric oxide synthase （NOS） in hypothalarnus and adrenal gland. METHODS： Ulcer index in rats and RT-PCR were used to study the protective effect of acupuncture on cold stress ulcer, and the expression of NOS in hypothalamus and adrenal gland. Images were analyzed with semi-quantitative method. RESULTS： The ulcer index significantly decreased in rats with stress ulcer. Plasma cortisol concentration was up regulated during cold stress, which could be depressed by pre-acupuncture. The expression of NOS1 in hypothallamus increased after acupuncture. The increased expression of NOS2 was related with stress ulcer, which could be decreased by acupuncture. The expression of NOS3 in hypothalamus was similar to NOS2, but the effect of acupuncture was limited. The expression of NOS2 and NOS3 in adrenal gland increased after cold stress, only the expression of NOS1 could be repressed with acupuncture. There was no NOS2 expression in adrenal gland in rats with stress ulcer. CONCLUSION： The protective effect of acupuncturing Tsusanli （ST36） on the expression of NOS in hypothalamus and adrenal gland can be achieved.

Polymorphisms of Epstein-Barr virus BHRF1 gene, a homologue of bcl-2

Background and Objective: EBV BamHI fragment H rightward open reading frame 1 (BHRF1), the Epstein-Barr virus (EBV) early gene, is structurally and functionally homologous to the oncogene bcl-2 and may play an important role in the development of EBV-associated tumors. To characterize the polymorphisms of BHRF1 in EBV-associated tumors, we analyzed the sequences of BHRF1 in isolates from nasopharyngeal carcinoma (NPC) and EBV-associated gastric carcinoma (EBVaGC) biopsies as well as throat washing (TW) samples from healthy donors. Methods: BHRF1 DNA sequences were analyzed by polymerase chain reaction (PCR) and sequencing for 39 NPC samples, 40 EBVaGC samples, and 53 EBV-positive TW samples from healthy donors. The variants of BHRF1 gene were classified according to the signature changes. The EBV types 1 and 2 at nuclear antigen (EBNA) 3C locus were determined by PCR. Results: Compared with EBV standard cell line B95-8, all isolates carried a silent mutation at amino acid (AA) 80 (nucleotide 54616 T→C), the AA88 L→V mutation was found in most isolates, and the AA79 V→L mutation in a few isolates. Other mutations were sporadically distributed. Based on the mutations at AA88 and AA79, 3 distinct variants of BHRF1 genes, designated as 79V88V, 79L88L, and 79V88L, were identified. The 79V88V was the most common variant. The distribution of the BHRF1 variants among the NPC, EBVaGC, and TW samples was not significant. The corresponding regions of bcl-2 homologues were conserved in all isolates except for 3 samples. The distribution of BHRF1 variants in type 1 and type 2 strains was significant different (P < 0.001, contingency coefficient was 0.554). Conclusions: The 79V88V is the dominant variant in NPC, EBVaGC, and TW samples from healthy donors and preferential linkages between BHRF1 and EBNA3C variants exist. Conserved BHRF1 in Bcl-2 homologous domains is helpful to remain the important role of BHRF1.

Cromakalin pretreatment affects mitochondrial structure and function in a rat model of ischemia/reperfusion injury

BACKGROUND：Mitochondrial structural changes and energy dysmetabolism frequently occur subsequent to cerebral ischemia.Adenosine triphosphate（ATP）-sensitive potassium channel openers exhibit protective effects on cerebral ischemia/reperfusion injury.OBJECTIVE：To validate the effects of cromakalin on mitochondrial structure and function in ischemic penumbra brain tissue in a rat model of middle cerebral artery occlusion（MCAO）.DESIGN,TIME AND SETTING：The present single-factor analysis of variance,randomized,controlled,animal experiment was performed at the Institute of Brain Science,Affiliated Hospital of Qingdao University Medical College between October 2007 and March 2008.MATERIALS：Forty male,Wistar rats were randomly divided into four groups,with 10 rats per group：sham-operated,MCAO,MCAO＋ATP-sensitive potassium channel opener（cromakalin）,and MCAO＋cromakalin＋ATP-sensitive potassium channel blocking agent（glibenclamide）.METHODS：Focal cerebral ischemia/reperfusion injury was induced by MCAO in all groups except the sham-operated group.The MCAO cromakalin group was administered 10 mg/kg cromakalin（i.p.） prior to MCAO induction.The MCAO＋cromakalin＋glibenclamide group received an injection of 10 mg/kg cromakalin（i.v.）,and subsequently an injection of 10 mg/kg cromakalin（i.p.） prior to MCAO induction.MAIN OUTCOME MEASURES：At 24 hours after cerebral ischemia/reperfusion injury,cellular apoptosis was detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate（dUTP） nick-end labeling technique.Cytochrome C expression was measured by immunohistochemistry.In addition,mitochondrial swelling,membrane fluidity,membrane phospholipid and malonaldehyde（MDA） contents,as well as Na^＋-K^＋-ATPase,Ca^2 ＋-ATPase,and superoxide dismutase（SOD） activities were determined.RESULTS：Compared with the sham-operated group,the three ischemia groups exhibited significantly elevated mitochondrial MDA content,reduced membrane phospholipid and ATP contents,down-regulated membrane fluidity,and reduced Na^＋-K^＋-ATPase,Ca^2＋-ATPase,and SOD activities（P 〈 0.05-0.01）.In the MCAO＋cromakalin group,the number of apoptotic cells decreased,and cytochrome C expression,as well as MDA content,were reduced.However,ATP content and Na^＋-K^＋-ATPase,Ca^2＋-ATPase,and SOD activities significantly increased compared with the MCAO group（P 〈 0.05-0.01）.Glibenclamide noticeably antagonized cromakalin protection of mitochondria.CONCLUSION：Pretreatment with the ATP-sensitive potassium channel opener cromakalin increased mitochondrial Na^＋-K^＋-ATPase,Ca2＋-ATPase,and SOD activities,decreased neuronal apoptosis,and inhibited cytochrome C expression following MCAO.

Ischemic stroke susceptibility gene in a Northern Han Chinese population

Interleukin-18 gene promoter polymorphisms are potential risk factors for ischemic cerebrovascular disease, and the –607C allele may increase ischemic stroke risk in the Han Chinese population. In the present study, we recruited 291 patients with ischemic cerebrovascular disease from the Affiliated Hospital of Qingdao University Medical College, China, and 226 healthy controls. Both patients and controls were from the Han population in northern China. Immunoresonance scattering assays detected increased serum amyloid A protein, C-reactive protein, and interleukin-18 levels in ischemic cerebrovascular disease patients compared with healthy controls. Analysis of the –607C/A （rs1946518） polymorphism in the interleukin-18 gene promoter showed ischemic cerebrovascular disease patients exhibited increased frequencies of the CC genotype and C alleles than healthy controls. Genotype and allele frequencies of the interleukin-18 –137G/C （rs187238） polymorphism and the –13T/C （rs11024595） polymorphism in the 5＇-flanking region of serum amyloid A, showed no significant difference between the two groups. Multivariate logistic regression analysis on the interleukin-18 promoter A/C genetic locus, for correction of age, gender, history of smoking, hypertension, diabetes mellitus, hypercholesteremia, and an ischemic stroke family history, showed ischemic cerebrovascular disease risk in individuals without the A allele （C homozygotes） was 2.2-fold greater than in A allele carriers. Overall, our findings suggest that the –13T/C （rs11024595） polymorphism in the 5′-flanking region of serum amyloid A has no correlation with ischemic cerebrovascular disease, but the C allele of the –607C/A （rs1946518） polymorphism in the interleukin-18 promoter is a high-risk factor for ischemic cerebrovascular disease in the Han population of northern China. In addition, the A allele is likely a protective gene for ischemic cerebrovascular disease.

Association of cholesterol with risk of pancreatic cancer:A meta-analysis

AIM: To evaluate the effect of dietary cholesterol and serum total cholesterol(TC) on the risk of pancreatic cancer. METHODS: A literature search was performed up to June 2014 in Pub Med, EMBASE, China National Knowledge Infrastructure and China Biology Medicalliterature database for relevant articles published in English or Chinese. Pooled relative risks(RRs) with 95% confidence intervals(CIs) were calculated with a random-effects model. RESULTS: We included 14 published articles with 439355 participants for dietary cholesterol, and 6 published articles with 1805697 participants for serum TC. For the highest vs lowest category of dietary cholesterol, the pooled RR(95%CI) of pancreatic cancer was 1.308(1.097-1.559). After excluding two studies(RR > 3.0), the pooled RR(95%CI) was 1.204(1.050-1.380). In subgroup analysis stratified by study design, the pooled RRs(95%CIs) were 1.523(1.226-1.893) for case-control studies and 1.023(0.871-1.200) for cohort studies. The association of dietary cholesterol with the risk of pancreatic cancer was significant for studies conducted in North America [1.275(1.058-1.537)] and others [2.495(1.565-3.977)], but not in Europe [1.149(0.863-1.531)]. No significant association [1.003(0.859-1.171)] was found between the risk of pancreatic cancer and serum TC. CONCLUSION: Dietary cholesterol may be associated with an increased risk of pancreatic cancer in worldwide populations, except for Europeans. The results need to be confirmed further.