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.

Objective:To analyze the inhibiting mechanism of microRNA-199a/b-3p(miR-199)on cell proliferation of triple negative breast cancer(TNBC)cells.Methods:Expression of miR199 in BT549 and MDA-MB-231 cells transfected with miR-199a/b-3p was detected by qRT-PCR.The proliferation of BT549 and MDA-MB-231 cells transfected with miR-199 were analysed by MTT as-say.Cell cycle of TNBC cells transfected with miR-199 was detected by Flow Cytometry assay.Results:MiR-199a/b-3p could suppress the proliferation of BT549 and MDA-MB-231 cells.Comparing with normal control,the proliferation rate were up to(41.02±2.34)%and(28.42 ±6.70 )%,and the cell cycle were arrest at G 1 phase.Conclusion: MiR-199a/b-3p could suppress the proliferation of TNBC,and may be a promising anti-cancer drug for TNBC.

Objective:To probe the expression of miR-126 and VEGF in breast cancer, and the anti-tumor effect of miR-126. Methods:The expression of miR-126 and VEGF in breast cancer tissues and cells were detected by qRT-PCR and Western blot;after transfection with miR-126 mimics into MDA-MB-231,expression of VEGF was detected again,MTT assay and cell scratch test were used to verify the influence of miR-126 on proliferation and migration of tumor cells. Results: The expression of miR-126 was lower in the breast cancer tissues and cells,the expression of VEGF was negative correlation with it,increasing the expression of miR-126 may decrease the expression of VEGF and inhibit the proliferation and migration of breast cancer cells. Conclusion:miR-126 can reduce the proliferation and migration of breast cancer cells by inhibiting the expression of VEGF,which play an anti-tumor effect.

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 observe the inhibitory effect of haploid vaccine pcDNA-CEA625-667 and three tandem repeats of minigene DNA vaccine pcDNA-triCEA625-667 derived from CEA gene on tumor in mice bearing tumor and the changes of survival time. Methods:The experimental animal model of mouse liver cell carcinoma was established and the mice were immunized with pcDNA-CEA625-667 and three series of DNA vaccine. Some of the mice were treated with normal saline as control group. The growth curve of tumor growth curve was recorded and the effect of vaccine on the survival time of tumor bearing mice was observed. Results:Compared with the normal saline control group,the two vaccines were able to significantly inhibit the tumor size and growth rate ( P<0. 01 ) of CEA positive tumor bearing mice,the inhibition of pcDNA-triCEA625-667 vaccine group was significantly better than the pcDNA-CEA625-667 vaccine group (P<0. 01),while the two were not inhibited tumor growth in CEA negative tumor bearing mice. The average survival time of the pcDNA-CEA625-667 vaccine group was(48. 50±6. 73)d,and there was significant difference (P<0. 01) compared with the saline control group ( 39. 00 ± 6. 64 ) d. The survival time ( 48. 50 ± 6. 73 ) d of the pcDNA-triCEA625-667 vaccine group was significantly higher than that of the normal saline control group and the pcDNA-CEA625-667 vaccine group (P<0. 01). The survival time of CEA negative tumor bearing mice could not be prolonged in the two groups. Conclusion:Either the haploid or the three series of the DNA vaccine,were able to significantly inhibit tumor growth rate (P<0. 01) and significantly prolong the survival time (P<0. 01) of CEA positive tumor bearing mice,but they had no therapeutic effect on CEA negative tumor bearing mice.

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.

Objective:To observe the immunological activity of haploid vaccine and three tandem repeats of minigene DNA vac -cine derived from Carcinoembryonic Antigen (CEA) gene.Methods:The immunoreaction was induced by intramuscular injection with pc-DNA3.0,pcDNA-CEA625-667 and pcDNA-triCEA625-667 in BALB/c.Four weeks after injection,the spleen cells and serum were separa-ted respectively from the mice for the in vitro assessment .Changes of the T lymphocytes subset was analyzed by flow cytometry .Lymph proliferation responses were tested by 3 H-TdR incorporation ,IFN-γ,IL-4 and GM-CSF in their cultural supernatants were detected with ELISA and seral IgG antibody against CEA were detected with Western blot and ELISA .Results:The difference of the ratio of CD 4+/CD8+of the mice immuned by pc-DNA3.0,pcDNA-CEA625-667 or pcDNA-triCEA625-667 was not significant.Lymph proliferation responses were more significant in the mice immuned by pcDNA-CEA625-667 and pcDNA-triCEA625-667 in a shorter time by contrast with na ?ve mice.Low tilter IgG antibody against CEA was detected in the antiserum of the mice immuned by repeats of minigene DNA vaccine , which suggested the activation of helper T-cell.ELISA showed that the level of IFNγin the 3 days culture of the splenocytes was rela-tively higher in the groups of minigene DNA vaccination than in the control groups ,while IL-4 expression was absent in all groups .The immune response level elicited by three tandem repeats of minigene DNA vaccine pcDNA -triCEA625-667 was superior to that elicited by pcDNA-CEA625-667 ,which showed that any immunogenic inadequacies in minigene presentation can be rectified by linking itself in a string-of-beads vaccine.Conclusion:The haploid vaccine and three tandem repeats of minigene DNA vaccine derived from CEA geneboth can not change the ratio of CD 4+/CD8+but can induce the activation of helper T-cell and skew T -cells toward Th -1 response.The immune response level elicited by three tandem repeats of minigene DNA vaccine was superior to thatelicited by haploid vaccine .