Objective To investigate the effects of insulin resistance (IR)on hypoxia/reoxygenation (H/R)inj ury in PC1 2 cells and the protective effects of N-actyl-L-cystine (NAC)against H/R inj ury. Methods PC12 cells were treated with 100 nmol/L insulin to induce IR. The H/R injury model was established by Na2 S2 O4. CCK-8 assay was used to detect the cell viability. Glucose consumption was detected by glucose oxidase method. Activity of superoxide dismutase (SOD)was detected by xanthine oxidase method. The levels of malonaldehyde (MDA)were measured by thiobarbituric acid method. Flow cytometry was used to determine the apoptosis and mitochondrial membrane potential (MMP). Results High-insulin inhibited insulin-induced glucose uptake in PC12 cells without affecting the cell viability (P<0. 05). PC12 cells with IR exhibited lower SOD activity and higher levels of MDA (P<0. 05 ),and enhanced apoptosis and depolarization of MMP induced by H/R(P<0. 05). NAC neutralized these effects induced by IR(P<0. 05). Conclusion IR aggravates H/R injury in PC12 cells by enhancing oxidative stress and NAC reduces the H/R injury in PC12 cells with IR via inhibiting oxidative stress and stabilizing MMP.

Objective:To investigate the expression of microRNA-199a/b-3p (miR-199a/b-3p) in hepatocellular carcinoma ( HCC) tissues,and to explore the relationship with clinical outcomes. Methods: Real time quantitative PCR technique was used to measure the expression of miRNA-199a/b-3p in HCC tissues. The correlation between miR-199a/b-3p expression and the clinic pathological features of patients were analyzed. Results: Comparing with adjacent control, miRNA-199a/b-3p presented lower expressions in HCC tissues (P<0. 05);lower miR-199a/b-3p was found correlated with metastasis and poor survival. Conclusion:MiR-199a/b-3p take a crucial role in HCC metastasis and recurrence.

Objective:To analyze the effect of PAK4 interruption by microRNA-199a/b-3p (miR-199a/b-3p) on migration and invasion of hepatocellular carcinoma (HCC).Methods: To test targeting of PAK4 by miR-199a/b-3p,we used luciferase assay in HEK293T cells cotransfected miR-199a/b-3p mimcs and pmirGLO-PAK4 3′UTR.The expression of PAK4 in SMMC-7721 transfected with miR-199a/b-3p was detected by Western blot.The biology behaviors of SMMC-7721 cells transfected with miR-199a/b-3p or PAK4 Si were analysed by cell migration assay and invasion assay.Results:MiR-199a/b-3p could suppress the mRNA and protein ex-pression of PAK4 by targeting PAK4 3′UTR,and the downregulating PAK 4 expression suppress the migration and invasion of SMMC-7721 cells.Conclusion: MiR-199a/b-3p could suppress the expression of PAK 4, which are considered key HCC suppressors and inhibit the migration and invasion of HCC cells.

Objective:To analyze the inhibiting mechanism of microRNA-199a/b-3p ( miR-199a/b-3p) on cell motility of breast cancer cells.Methods:The expression of PAK4 in MDA-MB-231 cells transfected with miR-199a/b-3p was detected by Western blot.The biology behaviors of MDA-MB-231 cells transfected with miR-199a/b-3p or PAK4 SiRNA were analysed by cell migration assay,invasion assay and protrusion dynamics.Results: MiR-199a/b-3p could suppress the expression of PAK 4 in MDA-MB-231 cells.Comparing with normal control ,miR-199a/b-3p or PAK4 SiRNA could suppress the migration ,invasion and membrane protrusion of MDA-MB-231 cells.Conclusion:miR-199a/b-3p could suppress the expression of PAK4,which are considered key breast cancer suppressors and inhibit the cell motility of breast cancer cells.

Objective:To analyze the expression levels of serum microRNA-199a/b-3p ( miR-199-3p) in hepatocellular carcinoma (HCC) patients and to explore whether miR-199a/b-3p could be a novel biomarker of early HCC.Methods: Patients with normal liver,liver cirrhosis and early HCC in clinic were included in this study.Blood samples of the patients were collected for analy-sis.Real time quantitative PCR technique was used to measure the expression level of serum miRNA-199a/b-3p.The AFP expression was got from clinical data.Results: Comparing with normal liver and liver cirrhosis patients, miRNA-199a/b-3p presented lower expressions in early HCC patients (P<0.05);the accuracy and positive rate of serum miR-199a/b-3p detection in cirrhosis and HCC were better than AFP detection.Conclusion:Data indicated that miRNA-199a/b-3p had diagnostic value for indicating liver cirrhosis and early HCC.

AIM: To investigate the effect of suberoylanilide hydroxamic acid (SAHA) on the apoptosis of hu-man small-cell lung cancer H446 cells and its possible mechanism.METHODS: H446 cells were incubated in the medi-um containing SAHA.CCK-8 assay was used to detect the anti-tumor effect of SAHA on the H446 cells, and IC50 values of SAHA were calculated.Flow cytometry was used to analyze the apoptosis.After Notch3 gene was silenced, the pro-apopto-tic effect of SAHA on the H446 cells was inhibited ( P <0.05).Eukaryotic expression plasmid containing N3ICD was transfected into the H446 cells, so that N3ICD was expressed in the H446 cells.The mRNA expression of Notch3 was measured by RT-PCR.The protein levels of Notch3, N3ICD, Puma and cleaved caspase-3 were determined by Western blot.RESULTS: SAHA remarkably reduced the cell viability in a dose-dependent manner (P <0.05), and the IC50 value of SAHA was 1.91 μmol/L.SAHA induced apoptosis in a dose-dependent manner (P <0.05).The expression of Notch3 gene was negative in the H446 cells, SAHA reactivated Notch3 gene and Notch3 pathway in a dose-dependent manner (P <0.05).Notch3 knockdown inhibited apoptosis induced by SAHA (P <0.05).Over-expression of N3ICD up-regula-ted the protein levels of Puma and cleaved caspase-3.CONCLUSION: SAHA induces apoptosis in human small-cell lung cancer H446 cells by activating Notch3 pathway and up-regulating the protein level of Puma.

Objective To investigate the relationship of programmed cell death 4(PDCD4) with the invasion of astrocytic glio-mas .Methods Using the immunohistochemical method to detect the expression of PDCD4 in astrocytic gliomas in different grades . Measuring the peritumoral low-density area on MRI scan ,then compared with the results of immunohistochemical expression .Re-sults The downregulation of PDCD4 was with the increasing of the malignant grade of astrocytic gliomas .The tumor grade malig-nancy was positively correlated with the grade of the peritumoral low-density area on MRI scan(P<0 .05) ,while the expression of PDCD4 was negatively correlated with the grade of astrocytic gliomas (P<0 .01) .Conclusion PDCD4 might serve as one of the in-dicators of invasion and malignant phenotype for astrocytic gliomas .

Objective:To observe the effect of tannic acid on renal morphology and function of diabetes mellitus model rats ,and to explore the mechanism of improving effect of tannic acid from oxidative stress , nitrosative stress angle.Methods: 8 rats were randomly selected from 68 6-week-old male Wistar rats as normal control group and the remaining 60 rats accepted high-sugar and high-fat diet for 4 weeks, then were injected streptozotocin ( STZ, 52 mg/kg ) intraperitoneally in order to manufacture a diabetic rat model.Further the diabetic rats were randomly divided into model group ,aminoguanidine group ,low-dose of tannic acid group and high dose of tannic acid group.The rats in aminoguanidine group were injected aminoguanidine [AG,40 mg/(kg· d)] intraperitoneally, those in low-dose of tannic acid group were injected tannic acid [TA,20 mg/(kg· d)] and those in high-dose of tannic acid group were injected tannic acid [TA,30 mg/(kg· d)].The rats in normal control group and model group were injected normal saline [NS, 30 mg/(kg· d)] and all rats were sacrificed and tissues were derived at the end of the week 10.Morphologic changes of kidney in diabetic rats were observed by HE staining and correlative biochemical indices of renal function were detected by biochemical analyzer.8-hydroxy deoxyguanosine (8-OHdG) and 3-nitrotyrosine (3-NT) content of renal tissue in rats in different groups were detected by ELISA method.Mesangial cells cultured in vitro were treated with high concentration of glucose (30 mmol/L) and AGEs (250 mg/L) and at the same time with different concentration of tannic acid (10,20,40 and 80μmol/L) on the basis of setting corre-sponding control group.The contents of 8-OHdG and 3-NT in the culture supernatant were measured by ELISA method after 48 hours.Results:Tannic acid can effectively improve the renal pathological changes and improve renal function of diabetic rats .The contents of 8-OHdG and 3-NT in kidney tissue homogenate of diabetic rats and in the supernatant of GMC cultured with high glucose or AGEs were all significantly increased and can be reduced by tannic acid.Conclusion:Tannic acid improving the structure and function damage of kidney in diabetic rats might be achieved by oxidative stress and nitrosative stress mechanism .

Objective:To research rmIL-18 in vitro culture system CCs induce tumor-specific cytotoxic T lymphocytes CTL and anti-tumor effect in mice.Methods:Used Stem SepTM immune magnetic cells separation method to culture mouse spleen NK cells,T cells and DCs,established culture systems in vitro;used of different approaches,different doses rmIL-18 to immunize HCC tumor-bearing mice,researched the effect of rmIL-18 on tumor growth rate and survival time.Results:rmIL-18 could induce and promote tumor-specific CTL-mediated killing effects in vitro culture system;tumor-specific CTL could significantly inhibit tumor growth(P<0.01) of and prolong the survival time of liver cancer tumor-bearing mice(P<0.01),and the effect was increased with rmIL-18 concentration increased(P<0.01),and intratumoral injection was superior to intraperitoneal injection(P<0.01).Conclusion:rmIL-18 can induce tumor-specific CTL in vitro and play a role in anti-liver cancer in mice.

Combining the functional metabolic imaging and anatomical imaging,PET-CT stands for the highest level of current nuclear medical imaging techniques,and has been applying increasingly in acupuncture studies.However,owning to complexity of the brain function and sensitivity of brain metabolism,the lower reproducibility of cerebral function imaging even may reverse many results.This has been provoked more and more attention by researchers.Its main cause is intimately related to poor experimental methodology.For this reason,the present paper raises the quality control of the research on mechanisms of acupuncture in the process of application of PET-CT techniques from 1) the included standards of participants,2) the preparation of participants during the scanning process of PET-CT,3) the setting of the scanning parameters,4) the used machine and the imaging reagent,5) the analysis of dada,6) the standardization of acupuncture manipulation,and 7) the designs of the acupuncture operation,needling opportunity and scanning opportunity,hoping to offer some beneficial information for the coming researches.