Expression of c-erbB-2 oncogene protein, epidermal growth factor receptor, and TGF-β1 in human pancreatic ductal adenocarcinoma

Objective: To detect the relations of c-erbB-2 onco-gene protein, epidermal growth factor receptor (EG-FR) and transforming growth factor-β1 (TGF-β1)to the progression or metastasis of pancreatic carci-noma.Methods: Using streptavidinbiotin complex (SABC)method, c-erbB-2 oncongene protein, we examinedimmunohistochemically EGFR and TGF-β1 expres-sions in wax-tissue sections from 10 individuals withnormal pancreas (NP), 13 patients with chronic pan-creatitis (CP) and 36 patients with pancreatic ductaladenocarcinoma (PC).Results: The positive expression rates of c-cerbB-2oncogene protein, EGFR and TGF-β1 in the NP, CPand PC groups were 0, 0, 10%; 7.7%, 7.7%,7.7%; and 41.7%, 50.0%, 44.4%, respectively.The positive expression rates of the three specific pro-teins increased more significantly in the PC groupthan in the NP and CP groups (P【0.05). The indi-vidual expression of c-erbB-2, EGFR and TGF-β1was not related to the age and sex of the patients aswell as the site, size and histopathological grade oftumors (P】0.05), but to the clinical stage of tumors(P【0.01). The coexpression rate of the three pro-teins was 27.8 % (10/36). This coexpression in thePC group was correlated with the histopathologicalgrades and clinical stages of tumors (P【0.01).Conclusion: Detection of c-erbB-2 oncogene protein,EGFR, and TGF-β1 expressions in pancreatic tissueis helpful to judge the malignancy, progression, andmetastasis of PC.

Response of Subcutaneous Xenografts of Endometrial Cancer in Nude Mice to Inhibitors of Phosphatidylinositol 3-Kinase/Akt and Mitogen-Activated Protein Kinase (MAPK) Pathways: An Effective Therapeutic Strategy for Endometrial Cancer

Objective: This study was designed to explore whether inhibition of the extracellular-regulated kinase (ERK) and phosphatidylinositol-3-kinase (PI3K) signaling pathways can inhibit the growth of xenografts of endometrial cancer cell lines with different estrogen receptors (ER) profiles in vivo and to provide preliminary laboratory basis for the probability of endometrial adenocarcinoma treatment with blockage of the two pathways, especially to endometrial cancer with low ER status. Methods: Human endometrial cancer Ishikawa bearing ER and HEC-1Awith low ER status cells were subcutaneously injected into BALB/c nude mice to establish endometrial cancer xenograft tumor models. The effects of PI3K/Akt inhibitor LY294002, MAPK/ERK1/2 inhibitor PD-98059 and their combinations on the growth of the xenograft tumors and apoptotic state of Ishikawa and HEC-1Acells were tested in vivo using the inhibitory rate, the terminal deoxynucleotidyl transferase-mediated nick-end labeling assay, H/E-stain. Western blot analysis was used to detect the alterations of activated ERK (P-ERK) and AKT (P-AKT) during this process. Results: LY294002, a PI3K/Akt pathway inhibitor, induced significant suppression in the growth of both Ishikawa and HEC-1Acell xenograft tumors, concomitant with increased apoptosis in xenografts as evidenced by TUNEL. A similar effect was also observed when the MAPK/ERK1/2 signaling pathway was inhibited by PD98059. Concurrent inhibition of the PI3K/Akt and MAPK/ERK1/2 pathways showed enhanced anti-tumor effects in vivo as indicated by increased apoptosis. At the same time, the levels of P-ERK and P-AKT in both xenograft tumors decreased, and their levels in combination group was the lowest. Conclusions: PD98059, LY294002 and their combinations showed remarkable inhibitory effects on xenograft tumors of endometrial carcinoma cell lines with different expression status of ER in vivo through blockage of PI3K/Akt and MAPK/ERK1/2 signaling pathways. This suggests that targeting these pathways may be an effective therapeutic strategy against endometrial carcinomas, especially for ER-negative cancers which show poor response to endocrinal therapy.

Epidermal growth factor receptor and K-Ras in non-small cell lung cancer-molecular pathways involved and targeted therapies

Lung cancer is currently the leading cause of cancer death in Western nations.Non-small cell lung cancer(NSCLC)represents 80%of all lung cancers,and adenocarcinoma is the predominant histological type.Despite the intensive research carried out on this field and therapeutic advances,the overall prognosis of these patients remains unsatisfactory,with a 5-year overall survival rate of less than 15%.Nowadays,pharmacogenetics and pharmacogenomics represent the key to successful treatment.Recent studies suggest the existence of two distinct molecular pathways in the carcinogenesis of lung adenocarcinoma:one associated with smoking and activation of the K-Ras oncogene and the other not associated with smoking and activation of the epidermal growth factor receptor(EGFR).The K-ras mutation is mainly responsible for primary resistance to new molecules which inhibit tyrosine kinase EGFR(erlotinib and gefitinib)and most of the EGFR mutations are responsible for increased tumor sensitivity to these drugs.This article aims to conduct a systematic review of the literature regarding the molecular pathways involving the EGFR,K-Ras and EGFR targeted therapies in NSCLC tumor behavior.

Transretinoic acid inhibits rats gastric epithelial dysplasia induced by N-methyi-N-nitro-N-nitrosoguanidine:influences on cell apoptosis and expression of its regulatory genes

INTRODUCTIONGastric epithelial dysplasia (GED) hypothetically is a straight-forward concept: dysplastic epithelium replacing the normal gastric epithelium of the stomach [1].In the stomach ,like any other segment of the gut ,it is defined as an unequivocal non-invasive epithelial change[2,3].The observation of gastric dysplasia as a cancerous lesion was recognized over a century ago ,but it is only after the advent of gastroscopy that its clinical significance has been stressed[4-7].

Gastrointestinal tumors in transplantation: Two case reports and review of literature

BACKGROUND Gastrointestinal stromal tumors(GISTs)are the most common mesenchymal tumors of the gastrointestinal tract.As most of them harbor a KIT mutation(75%),selective kinase inhibitors are the therapeutic option and show a sustained objective response among patients with metastatic or unresectable GISTs.A wellknown higher risk of neoplasm has been described among renal transplant recipients(RTRs).Nevertheless,only few cases of GIST onset among transplant patients have been reported in the literature.CASE SUMMARY Here,we describe 2 cases of gastric GIST occurring during the follow-up of RTRs.We also review the existing literature concerning GIST occurrence in transplant patients.In total and in association with our 2 cases,16 patients have been reported.The median age was 59.5 years and 69%were male.With a median tumor size of 45 mm,no patient displayed metastatic dissemination at diagnosis.Time from transplantation to diagnosis was highly variable between 5 mo and 21 years.Histopathological data mostly revealed high risk of progression(43%).Death increased to 29%during follow-up.Surgical treatment was systematically performed when the tumor was operable(94%).The use of adjuvant therapy was uncommon(19%).CONCLUSION GISTs represent rare but potentially severe malignant complication among transplant patients.

Advances in BRAF gene mutations in papillary thyroid carcinoma

Thyroid cancer is the most common endocrine system tumor.Ultrasound guided fine needle puncture(FNA)can identify benign and malignant thyroid nodules.However,due to the limitation of cytological detection,some thyroid nodules are difficult to distinguish benign and malignant.BRAF gene mutation is a common human oncogenic mutation and the highest mutation frequency in papillary thyroid carcinoma.The combination of FNA and BRAF gene detection can significantly improve the diagnostic rate of benign and malignant thyroid nodules and make up for the deficiency of single diagnosis of cytology.Moreover,while the incidence of thyroid cancer is growing rapidly worldwide,its mortality remains stable.The problem of overdiagnosis and overtreatment of thyroid cancer is becoming more and more obvious.However,due to the limitations of current studies on BRAF genes,its prognostic value for papillary thyroid carcinoma remains controversial.Therefore,in order to reduce the adverse effects of overdiagnosis and treatment,the relationship between gene and tumor biological behavior needs further study in the future.

Efficacy of Ganshuang granules(肝爽颗粒)on non-alcoholic fatty liver and underlying mechanism:a network pharmacology and experimental verification

OBJECTIVE:To investigate the potential pharmacological mechanisms of Ganshuang granules(肝爽颗粒,GSG)in treating non-alcoholic fatty liver(NAFLD).METHODS:All the active components and targets of GSG were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.Protein-Protein interaction network,Kyoto Encyclopedia of Genes and Genomes and Gene Ontology function annotation of common targets were analyzed to predict the mechanisms of action of GSG in the treatment of NAFLD.Then,the mouse models of NAFLD were constructed in a diet-induced manner and treated with GSG.The levels of interleukin 6(IL-6),tumor necrosis factor-alpha(TNF-α)and phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT)pathway-related proteins in the liver of mice in each group were measured by enzyme linked immunosorbent assay and Western blot,respectively.RESULTS:Network pharmacology revealed a total of 159 potential targets of GSG for the treatment of NAFLD.Functional enrichment analysis indicated that the PI3K/AKT signaling pathway may be involved during GSG treatment of NAFLD.Further experiments showed that the significantly decreased alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase,total cholesterol,triglyceride and low-density lipoprotein cholesterol levels in NAFLD model mice serum after GSG treatment,as well as the expression levels of IL-6 and TNF-αin the liver.Furthermore,drug intervention increased the protein expression levels of phosphorylated-PI3K(P-PI3K)and P-AKT in the liver of the model group mice,and decreased the protein expression level of sterol regulatory element-binding protein 1.CONCLUSION:We found that GSG is effective in treating NAFLD and the potential therapeutic targets may be involved in PI3K/AKT signaling pathway.

Mechanism of total glucosides from Chishao(Radix Paeoniae Rubra)on proliferation and apoptosis of hepatocellular carcinoma cells via phosphatase and tensin homolog deleted on chromosome ten/phosphatidylinositol 3-kinase/protein kinase B signaling pathway

OBJECTIVE:To investigate the possible molecular mechanism of total glycosides of Chishao(Radix Paeoniae Rubra)(TG-RPR)on proliferation and apoptosis of hepatocellular carcinoma cells.METHODS:The proliferation of TG-RPR on Hep G2 cells was detected using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.The apoptosis of Hep G2 cells was measured by annexin V-FITC/double staining.The phosphatase and tensin homolog deleted on chromosome ten(PTEN)/phosphatidylinositol 3-kinase(PI3 K)/protein kinase B(Akt)signaling pathway was evaluated by Western Blot and reverse transcription-polymerase chain reaction(RT-PCR).RESULTS:TG-RPR can up-regulation the expression of pro-apoptotic factors such as PTEN and BCL2-Associated X(Bax),down-regulation the expression of anti-apoptotic factors including B-cell lymphoma-2(Bcl-2),PI3 K,and Akt.CONCLUSION:TG-RPR significantly inhibits the proliferation of Hep G2 cells in a dose-dependent manner and promotes apoptosis.These results demonstrated TG-RPR has significant inhibitory effect on Hep G2 cells.These results identify a critical role of TG-RPR in proliferation and apoptosis of Hep G2 cells via modulating PTEN/PI3 K/Akt signaling pathway.TG-RPR may offer a promise as a potential pharmaceutical therapy for hepatocellular carcinoma.

肝细胞生长因子／c—met系统与头颈肿瘤

肝细胞生长因子是一种多功能的细胞生长因子，其生物活性由c—met蛋白所介导，肝细胞生长因子通过与原癌基因c-met受体结合激活受体酪氨酸蛋白激酶活性调节细胞的分裂、增殖、分化、迁移等。故肝细胞生长因子／c—met信号传导在肿瘤的侵袭转移中发挥重要作用。人类许多肿瘤的生长、侵袭和转移与肝细胞生长因子／c—met系统密切相关。研究肝细胞生长因子／c-met系统与头颈部肿瘤侵袭、转移的关系，将对头颈肿瘤的诊断、治疗及预后有重要意义。

甲状腺髓样癌研究进展

甲状腺髓样癌（medullary thyroid carcinoma,MTC）来源于分泌降钙素和癌胚抗原的甲状腺滤泡旁细胞（又称C细胞）,且多因RET基因突变引发.它属于甲状腺癌中恶性程度较高的一类,预后相对较差.本文重点综述了近年来甲状腺髓样癌基础分子生物学研究、诊断、影像学检查及治疗方面的进展.

Effects of electroacupuncture at Taichong(LR 3) and Baihui(DU 20)on cardiac hypertrophy in rats with spontaneous hypertension

OBJECTIVE: To investigate the effects of electroacupuncture(EA) at Taichong(LR 3) and Baihui(DU 20)on myocardial hypertrophy in spontaneously hypertensive rats(SHRs).METHODS: Thirty-six SHRs were randomly assigned to model, EA, and Losartan groups, with twelve rats per group. Twelve Wistar Kyoto rats were selected as the normal control group. Systolic blood pressure(SBP) and cardiac function were measured in all rats.Expression levels of factors associated with the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR) pathway were evaluated by Western blotting and real-time PCR.Pathological changes of the heart tissue were observed by hematoxylin-eosin staining.RESULTS: After treatment, enhanced SBP was significantly decreased in the EA and Losartan groups compared with the model group(P < 0.01). Echocardiographic and morphological analyses revealed that enhanced end-diastolic interventricular septal thickness and left ventricular posterior wall thickness, as well as ratio of left ventricular weight to body weight were markedly diminished in the EA and Losartan groups(P < 0.01 or P < 0.05), while reduced left ventricular end-diastolic dimension and left ventricular ejection fraction were significantly ameliorated(P < 0.01). Real-time PCR and western blotting analyses showed that the expression levels of PI3K,Akt, and mT OR in SHRs were significantly up-regulated by EA and Losartan(P < 0.01), while the expression levels of PTEN and ANP were down-regulated(P < 0.01).CONCLUSION: EA at Taichong(LR 3) and Baihui(DU20) inhibited the development of cardiac hypertrophy and improved the cardiac function in SHRs, possibly through regulation of the PI3K/Akt/mTOR signalling pathway.

Overexpression of Bcl-2 partly inhibits apoptosis of human cervical cancer SiHa cells induced by arsenic trioxide

OBJECTIVE: To study the biological effect of arsenic trioxide (As2O3) on human cervical cancer SiHa cells and SiHa cells overexpressing bcl-2 gene. METHODS: SiHa cells with overexpression of Bcl-2 (SiHa-Bcl2 cells) were established by transfecting SiHa cells with Bcl-2 expression vector. The sensitivities of SiHa and SiHa-Bcl2 cells to As2O3 were determined using MTT (Thiazolyl blue) reduction and colony forming ability assay, morphological analysis, flow cytometric analysis, DNA agarose gel electrophoresis, in situ cell death detection (TUNEL), Northern blot, RT-PCR and Western blot. RESULTS: As2O3 inhibited the growth of SiHa cells and induced G2/M arrest and apoptosis of the cells. RT-PCR and Western blot analysis revealed that As2O3 induced SiHa cell apoptosis possibly via inhibiting the expression of HPV16 E7 and decreasing the expression of c-myc. However, we found that SiHa-Bcl2 cells partly resisted As2O3 induced apoptosis, which might be related to the prevention of the down-regulation of HPV16 E7 and c-myc gene expression. Nevertheless, As2O3 at a high concentration could still induce apoptosis of SiHa-Bcl2 cells mainly via decreasing Bcl-2 expression and slightly inhibiting viral gene expression. CONCLUSION: As2O3 is an inducer of the apoptosis of human cervical carcinoma cells and the cells overexpressing Bcl-2 can partly resist As2O3 induced apoptosis, but the exact mechanism is unclear.

Effects of External Application of Chinese Medicine on Diabetic Ulcers and the Expressions of β-catenin,c-myc and K6

Objective：To observe the clinical efficacy of Chinese medicine（CM） treatment of Hongyou Ointment（红油膏） and Shengji Powder（生肌散） on diabetic ulcers,and to observe the influence of CM treatment on the expressions of proteins associated with the Wnt signaling pathway,such asβ-catenin,c-myc and K6.Methods：sixty-two patients fitting the registration standards were randomly divided into the CM group（31 patients） and the Western medicine（WM） group（31 patients） by a random number table.The patients in the CM group were treated with Hongyou Ointment and Shengji Powder externally.The patients in the WM group were treated with mupirocin ointment,growth factor（bFGF）,and Vaseline gauze for external use and with basic therapies.Wound-healing time and four-week healing rate were recorded.The wounds were measured by digital photography and ImageJ software.Skin biopsies were obtained from 24 patients before CM treatment and 20 patients after CM treatment.Immunohistochemical tests and semi-quantitative imaging with NIH ImageJ 1.42 software were used to analyze the changes in protein expression ofβ-catenin,c-myc,and K6.Results：Fifty-three patients completed the trial;four patients in the CM treatment group and five patients in the WM group dropped out.Among them,four were dissatisfied with the treatment process,two could not continue because of their jobs,two failed to complete the course of follow-up appointments,and one was diagnosed with squamous cell carcinoma during treatment.The comparison of ulcer healing rates between the two groups showed insignificant differences（P=0.77）.The ulcer healing rates were 33.33%（9/27） in the CM group and 26.92%（7/26） in the WM group.However,the effective rate was significantly higher in the CM group（81.48%,22/27） than in the WM group（57.69%,15/26,P=0.04）.The mean wound healing time was shorter in the CM group（22.71±5.46 days） than in the WM group（26.56±7.56 days,P=0.04）.CM treatment was well tolerated,and there was no withdrawal due to adverse reactions.Immunohistochemical analysis in the refractory wound indicated higher expressions ofβ-catenin,c-myc and K6 compared with the normal skin.β-catenin was abnormally expressed in the nuclei of the keratinocytes and fibroblasts at the wound margins,and the expressions of c-myc and K6 were highly expressed in the full hyperplastic epidermis,especially in the granular layer（P0.05）.The expressions of these proteins decreased after CM treatment.The expression levels ofβ-catenin,c-myc,and K6 proteins before and after the treatment were 101.88±10.76 vs.140.42±8.45;113.27±16.75 vs.153.79±8.32; 90.39±11.07 vs.151.29±7.39,respectively.Conclusions：CM treatment using Hongyou Ointment and Shengji Powder was efficient in the management of diabetic skin ulcers.The mechanism of action might be related to the inhibition of the Wnt signaling pathway.

Spleen-kidney supplementing formula alleviates insulin resistance via regulating AKT/glycogen synthase kinase 3β pathway in rats with type 2 diabetic induced by high-fat diet

OBJECTIVE: To explore the molecular mechanism underpinning the action by investigating its effect on glycogen content and AKT(also known as protein kinase B)/glycogen synthase kinase 3β(GSK-3β) pathway in the liver of rats with type 2 diabetic induced by high-fat diet.METHODS: The rat model of type 2 diabetes was induced by high-fat diet and multiple low-dose streptozotocin injection. Diabetic rats were divided into five groups: the model control group, the Metformin group, spleen-kidney supplementing formula groups of low, medium and high doses. Fasting blood glucose(FBG) levels were measured before treatment and every two weeks during treatment.After the treatment, oral glucose tolerance test was performed, and hemoglobin A1c (HbA1c) and C-peptide were measured to assess the formula's effect on glucose metabolism and insulin resistance. The protein expression levels of AKT, GSK-3βand their phosphorylated forms in the liver were also measured to study the formula's role in insulin signaling pathway.RESULTS: Spleen-kidney supplementing formula significantly relieved the symptoms of polydipsia,polyuria and weight loss in type 2 diabetic rats, reduced FBG and HbA1c levels, increased glycogen content, and improved insulin sensitivity. The anti-diabetic effects of spleen-kidney supplementing formula are dose dependent. It also increased the total AKT protein level and the GSK-3β phosphorylation in the liver of type 2 diabetic rats.CONCLUSION: Spleen-kidney supplementing formula has hypoglycemic effect and relieves insulin resistance by enhancing AKT/GSK-3β signaling pathway in the liver of type 2 diabetic rats.

Noni(Morinda citrifolia L.) fruit juice delays immunosenescence in the lymphocytes in lymph nodes of old F344 rats

Objective： Aging is associated with the development of diseases because of immunosuppression and altered functioning of the neuroendocrine system. The medicinal properties of Morinda citrifolia L. have been widely exploited for the treatment of age-associated diseases. This study aims to investigate the in vitro and in vivo effects of noni（M. citrifolia） fruit juice（NFJ） on neuro-immunomodulation in the lymph node lymphocytes of F344 rats.Methods： Lymphocytes isolated from axillary and inguinal lymph nodes of young（3–4 months） and old（18–21 months） rats were treated in vitro with different concentrations（0.0001%, 0.01%, and 1%） of NFJ for a period of 24 h. In the in vivo study, old（16–17 months） male F344 rats were treated with 5 mL/kg body weight of 5%, 10% and 20% of NFJ, twice a day, by oral gavage, and lymph node lymphocytes were isolated after 60 d. Concanavalin A（Con A）-induced lymphocyte proliferation, interleukin-2（IL-2）and interferon-γ（IFN-γ） production and expression of intracellular markers, such as phosphoextracellular signal-regulated kinase（p-ERK1/2）, phospho-γ AMP response element-binding protein,phospho-protein kinase B（p-Akt）, phospho-tyrosine hydroxylase（p-TH）, phospho-nuclear factor of κ light polypeptide gene enhancer in B-cells inhibitor-α（p-IκB-α） and phospho-nuclear factor-κB（p-NF-κB p65 and p50） were examined in the lymphocytes of lymph nodes.Results： NFJ increased Con A-induced lymphocyte proliferation, IL-2 and IFN-γ production, and p-ERK1/2 expression both in vitro and in vivo. In in vivo NFJ-treated old rats, lymph node lymphocytes showed increased expression of p-TH and Akt, nitric oxide production and decreased expression of p-NF-κB p65 and p50.Conclusion： These results suggest that the immunostimulatory properties of NFJ are facilitated through intracellular signaling pathways involving ERK1/2, Akt and NF-κB.

Detection of pim-1 mRNA in prostate cancer diagnosis

Background Pim-1 plays an important role in the apoptosis, proliferation, differentiation of cancer cells and progression of cancer. In this study we detected the expression of pim-1 mRNA in normal prostate, benign prostatic hyperplasia （BPH）, and prostate cancer （PCa） and explored its diagnostic value for PCa. Methods The prostate tissues were collected from 23 patients with PCa, 37 patients with BPH, and 3 healthy volunteers. Pim-1 mRNA expression levels in these samples were determined by the quantitative real-time PCR （QRT-PCR）. The differences of expression were calculated based on a standard curve. Results The ratio of pim-1 mRNA to β-actin in the normal prostate, BPH, and PCa were 1.05±0.04, 2.57±0.74 and 4.45±0.63, respectively. The differences among PCa, BPH and NT were significant （P〈0.05, respectively）. Conclusion Detecting pim-1 mRNA expression by QRT-PCR provides a reliable metric for the diagnosis of PCa.

Diazoxide preconditioning plus subsequent hypothermia increased resistance of rat cultured hippocampal neurons against hypoxia-reoxygenation injury

Background Cerebral ischemia-reperfusion/hypoxia-reoxygenation insult triggers lots of pathophysiological and biochemical events that separately affect the evolution of cerebral damage. Accordingly, all known effective neuroprotective measures should be taken to get the optimal efficacy of therapy. This study was undertaken to investigate whether diazoxide （DZ） preconditioning combined with the following hypothermia could contribute to synergistic neuroprotection compared with either hypothermia or DZ preconditioning alone. Methods Cultured for 9-10 days in vitro, the hippocampal neurons of SD rats were preconditioned with DZ 0 pmol/L or DZ 250 pmol/L for 1 hour per day and this treatment lasted for 3 days. Subsequently, neurons were subjected to deprivation of oxygen for 4 hours at 37°0, 34°C, 30℃ and 22℃, respectively. This experiment consisted of 8 groups （4 temperature groups and 4 combination groups） and each group contained 12-well or 2-dish cells. Survival rate, expression of Bcl-2, fluorescence magnitude of intracellular calcium, and concentration of malondialdehyde （MDA） were determined at 24 hours after reoxygenation. Results The survival rate and expression of Bcl-2 were both increased in individually hypothermic conditions compared with those at 370G （P〈0.05）, whereas intracellular calcium and MDA did the opposite exhibition simultaneously （P〈0.05）. 22℃ contributed to a higher survival rate and greater expression of Bcl-2 in comparison with other hypothermia （P〈0.05）. Preceding administration of 250 pmol/L DZ took the similar effects on the neurons like hypothermia. Moreover, compared with individual hypothermia or DZ preconditioning, the neuronal survival rate and expression of Bcl-2 in the combination group were increased significantly （P〈0.05）, whereas the calcium fluorescence density and concentration of MDA were reduced further （P〈0.05）. 250 Iamol/L DZ preconditioning combined with 22℃ provided a maximal neuroprotection. Conclusions Compared with either individual hypothermia or DZ preconditioning, the combination of both treatments conferred synergistic protection for cultured hippocampal neurons in vitro against hypoxia- reoxygenation insult.

Network pharmacology and molecular docking study on the effect of Kaempferol in treatment of metabolic associated fatty liver disease

OBJECTIVE: To study therapeutic effect of kaempferol on metabolic associated fatty liver disease(MAFLD) by network pharmacology and molecular docking combined with cell experiments. METHODS: First, use the Swiss Target Prediction database to predict the targets of kaempferol, and collect the targets of MAFLD through the Disgenet database and the Gene Cards database. Then, the common target of kaempferol and MAFLD was enriched and analyzed by the Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes, and the protein-protein interaction(PPI) network was constructed through the string database to obtain the key targets, and carry out molecular docking of key targets with kaempferol;In cell experiment, oleic acid induced steatosis in Hep G2 cells, which was intervened by kaempferol, the level of triglyceride(TG) was detected, the lipid deposition was observed by oil red O staining, and the protein expression was detected by Western blot. RESULTS: The results showed that there are 33 common targets for kaempferol and MAFLD. The biological process of GO is related to the regulation of protein kinase B, cell apoptosis, inflammatory factors, lipoxygenase, etc. Its action pathway is related to the phosphatidylinositol-3-kinase and protein kinase B(PI3K-AKT) signaling pathway, hypoxia-inducible factor 1 signaling pathway, forkhead box protein O signaling pathway, AMP-activated protein kinase signaling pathway, tumor necrosis factor signaling pathway, etc., the key targets are protein kinase B(AKT1), prostaglandin G/H synthase 2, matrix metalloproteinase-9, epidermal growth factor receptor, and the molecular docking of kaempferol with the four key targets shows good binding properties. Cell experiments show that kaempferol can reduce cell TG levels, reduce lipid deposition, increase the expression of PI3K, AKT, and beclin-1, and reduce the expression of caspase-3 and nuclear factor-kappa B. Kaempferol can treat MAFLD by regulating the PI3K-AKT signaling pathway to regulate cell autophagy, apoptosis, and inflammation. CONCLUSIONS: This study shows that kaempferol can regulate lipid metabolism, reduce apoptosis, regulate inflammation and autophagy in the fatty liver cell model. It reveals the therapeutic mechanism of kaempferol on MAFLD and provides a natural product candidate for the treatment of MAFLD.

Prognostic significance of clinicopathologic parameters in gastrointestinal stromal tumors:a study of 156 cases

The biological behavior of gastrointestinal stromal tumors(GISTs)are highly variable.To invest-igate the biological behavior of GIST,we collected 83 cases of gastric and 62 cases of small intestinal GIST from the Department of Pathology of the Chinese PLA General Hospital.The parameters include age,primary tumor location,tumor diameter,mitotic index,tumor necrosis,risk assessment,clinical stage and the c-kit exon 11 muta-tion.All these were analyzed in 105 cases along with the follow-up data and tested by log rank and COX hazard proportional model.We find that the average age of gast-ric GIST was 55.4 years.Of the 62 cases that were fol-lowed up,17 cases had metastasis or recurrence and the 5-year survival rate was(66.51±17.06)%.For the small intestinal GIST,the average age was 50.6 years and 43 cases were followed up.Of these,22 cases had meta-stasis or recurrence and the 5-year survival rate was(61.76±18.30)%.Small intestinal GIST was more fre-quently associated with metastasis and tumor relapse than gastric GIST(x^(2)=56.131,P=0.013).For gastric GIST,patients younger than 50 years(P=0.046),the advanced clinical stage(P=0.0001),the large tumor diameter(P=0.0001),a high mitotic index(P=0.0001),necrosis(P=0.0001)and a high risk grade(P=0.004)were all correlated with a lower survival rate.The COX hazard proportional model revealed that advanced clinical stage(P=0.001),large tumor size(P=0.001),a high mitotic index(P=0.002)and the high risk grade(P=0.018)indi-cated a poorer prognosis in gastric GIST.For small intest-inal GIST,necrosis(P=0.036)and advanced clinical stage(P=0.010)were associated with lower survival rates and the clinical stage was shown to be an independent prognostic indicator.A total of 25 cases harbored muta-tions in c-kit exon 11.The frequency of c-kit mutation was 32%and 22.5%for gastric and small intestinal GIST,respectively.In gastric GIST,the mutated c-kit was pre-dominant in patients older than 50 years of age.But in the small intestinal GIST,the mutated c-kit was predominant in the age group of 40-49 years.In conclusion,for gastric GIST,clinical stage,tumor size,mitotic index,and risk grade are the prognostic indicators.For small intestinal GIST,necrosis and clinical stage are the prognostic indi-cators.Small intestinal GIST are more aggressive than gastric GIST.The occurrence of c-kit mutation may cor-relate with the age of patients.