AIM: To develop a reliable primary cultured hepatocytes model in vitro for liver metastasis research. METHODS: Hepatocytes were isolated by a modification of the two-step collagenase perfusion method. The apoptosis and cell cycle of hepatocytes were measured with flow cytometry. The proliferation of hepatocytes was detected by SRB method. RESULTS: The viability and purity of hepatocytes were 90% and 95%,respectively. The result of flow cytometry analysis showed that there was little apoptosis in hepatocytes and most of hepatocytes were in G_0/G_1 phase. The proliferation and albumin-secreting function of hepatocyte cultured by low glucose DMEM and high glucose DMEM were higher than that of cultured by RPMI1640 during 1 to 6 day, but there was no significant different between low glucose DMEM group and high glucose DMEM group. CONCLUSION: Hepatocytes have higher purity and viability with the normal biological activity for about 6 days by this method and it may be a cell model for the study of liver metastasis in vitro.

Objective To study the CT manifestation of testicular endodermal sinus tumor in childhood.Methods Clinical and CT data in sixteen cases of testicular endodermal sinus tumor in children were retrospectively reviewed.Results The tumors appeared as solid masses on plain CT scan,the density of the tumor was homogeneous in 14 cases and heterogeneous in 2 cases.The masses were obviously heterogeneous enhancement after administration of contrast material in 12 cases.Serum ?-fetoprotein(AFP)level was elevated to more than 1000?g/L(normal:less than 20?g/L).Conclusion CT is a valuable in diagnosis of testicular endodermal sinus tumor in children.CT combined with serum AFP can improve the diagnosis of the disease.

Objective To explore the effect of hepatitis B virus(HBV) on the expression of complement 3 (C3) and complement 4 (C4) and its regulatory mechanism.Methods Differentially expressed genes between HepG2 and HepG2.2.15 cells was screened by gene chip,serum complement component 3 (C3) and 4 (C4) levels in patients with HBV infection and in healthy individuals were measured by Immunoturbidimetry,HBV infectious clone pHBV1.3 was transfected into HepG2 cells,and expression of C3 and C4 was measured by RT-PCR and Western blot.Results Expression of C3 and C4 mRNA was lower in HepG2.2.15 cells than in HepG2 cells,serum C3 and C4 levels was much lower in patients with chronic hepatitis B and hepatic carcinoma as compared to healthy individuals (P＜0.05 ).HBV could downregulate the expression of C3 and C4 at mRNA and protein levels.Conclusion HBV may inhibit the expression both in vivo and in vitro.

Objective The experiment was conducted to explore the influence of progesterone antagonist\|RU486 on the macrophages in the uteri of pregnant mice and its relevance to pregnancy. Methods On day 4 and day10 of gestation, each mouse was subcutaneously injected with 150mg/L of RU486 (treatment group) or the same volume of saline (control group). The macrophages were immunohistochemically identified in the uterus 12, 24, 36?h after injection. Results Injection of RU486 on day 4 gestation had completely blocked the implantation of blastocysts. A large number of macrophages were found in the endometrium, myometrium and blood vessel layer 12～36?h after RU486 injection, which was significantly higher than that of the control groups ( P

Objective To study curative efficacy of compound tanshin injection and acupuncture and moxibustion in treatment of peripheral nerve injury its effects on nerve function.Methods 110 patients of peripheral nerve injury whoreceived therapy from April 2014 to September 2014 in our hospital were selected as research objects,according to the treatment were divided into observation group and control group,observation group using curative efficacy of compound tanshin injection,the control group using compound tanshin injection and low frequency dc therapy.Observe clinical efficacy of the two groups after treatment,compared two groups before and after treatment can motor nerve and sensory nerve nerve conduction velocity,and to evaluate the basic function after treatment.Results After treatment,the observation group total effectiveness 94.5％,obviously higher than the control group total effectiveness 81.8％,significant difference(P< 0.05); Observation group after treatment of nerve function was 80.0％,significantly higher than the control group was 60.0％,significant difference(P<0.05); And after the treatment of motor nerve and sensory nerve conduction velocity observation group were significantly higher than the control group,significant difference(P<0.05).Conclusion Curative of compound tanshin injection and acupuncture and moxibustion in treatment of peripheral nerve injury,can significantly improve patients with nerve function,improve the effect of treatment,and no side effect,is worth popularizing in clinic.

AIM: To investigate the role of brainstem auditory evoked potential(BAEP) in the early diagnosis of diabetic central neuropathy (DCN) and to probe into the relationship between plasma nitric oxide (NO) and DCN. METHODS: Experimental diabetes(ED) rat model was used. BAEP and plasma NO were measured dynamically after ED modeling. RESULTS: The peak latency (PL) of Ⅰ, Ⅲ, Ⅳ and V and the interpeak latency (IPL) of I-III and III-V of BAEP in ED group prolonged after 4 weeks of ED modeling (P＜0.05). All PL and IPL of BAEP prolonged after 6 weeks、8 weeks and 10 weeks (P＜0.01). The plasma NO increased to the top in ED group at 2 weeks and then decreased gradually. There was negative correlation between plasma NO and PL and IPL of BAEP. CONCLUSION: BAEP was sensitive and valuable in the early diagnosis of DCN. Reduction of plasma NO may play a role in the pathogenesis of DCN.

AIM: To investigate the role of brainstem auditory evoked potential(BAEP) in the early diagnosis of diabetic central neuropathy (DCN) and to probe into the relationship between plasma nitric oxide (NO) and DCN. METHODS: Experimental diabetes(ED) rat model was used. BAEP and plasma NO were measured dynamically after ED modeling. RESULTS: The peak latency (PL) of Ⅰ, Ⅲ, Ⅳ and V and the interpeak latency (IPL) of I-III and III-V of BAEP in ED group prolonged after 4 weeks of ED modeling (P

AIM:To clarify the effects of high fat/cholesterol diet on lipid metabolism and atherogenesis in treble genes mutant mice.METHODS:ApoE-/-/LDLR-/-/Leprdb/db mice were generated by cross apolipoprotein E,lower density lipoprotein receptor gene knockout mice with leptin receptor gene spontaneous point mutants.The mice were fed with high fat/cholesterol diet from 22-day-old.The total plasma cholesterol(TC),triglyceride(TG)and glucose levels were measured and pathological changes of aorta intima and liver were analyzed.RESULTS:A significant elevated TC,TG and glucose levels in plasma with progress of time in young treble gene mutant mice were observed,which were higher than that in ApoE-/-/LDLR-/-and Leprdb/db mutants.At time of only 2 weeks after fed with high fat/cholesterol diet,TC and TG levels reached(106.75?3.40)mmol/L,(9.12?1.35)mmol/L,respectively in treble gene mutant mice,4.33-and 2.36-fold higher than those in treble genes mutants fed with normal chow diet.The levels were continuously increased until final experimental point.Intima of the aorta appeared with various injuries such as edema,desquamation of the endothelial cells,foam cell formation,rupture of IEL in local regions of root and arch areas of aorta at 2 weeks after fed with high fat/cholesterol diet.Microscopic pathological complex of significant local intima incrassation and fatty change of the liver were observed in the mutants that fed with high fat/cholesterol diet for 8 weeks.Injuries of aorta were severe than normal dietetic control group.CONCLUSION:High fat/cholesterol diet as a key dietary factor is significant aggravated lipid metabolism abnormity,promotes early damage of aorta and process of atherogenesis in the treble genes mutants.

Objective: To explore the effect of hepatitis C virus (HCV) on the expression of serine protease inhibitor Kazal1 (SPINK1) and its clinical implication. Methods: mRNA and protein expression of SPINK1 in Huh7.5.1 cells infected by HCV JFH-1 and the control cells were measured by RT-PCR and western blotting, SPINK1 levels in the cell supernatants and sera of HCV patients were measured by enzyme-linked immunosorbent assay (ELISA), the difference of SPINK1 levels between healthy controls and HCV patients was analyzed. Results: Expression of SPINK1 mRNA and protein was higher in Huh7.5.1 cells infected by HCV JFH-1 than in the control cells, serum SPINK1 levels was much higher in HCV patients than in healthy controls (P=0.016). Conclusions HCV can upregulate the expression of SPINK1.

OBJECTIVE: To assess the association of apolipoprotein (apo) C1 (APOC1) gene rs4420638A/G and -317H1/H2 polymorphisms with the risk of pre-eclampsia (PE) and the influence of their genotypes on the clinical and metabolic indexes among Chinese women. METHODS: In total 289 PE patients and 824 women with uncomplicated pregnancies were included. The rs4420638A/G genotype was determined by a Taqman real-time PCR allelic discrimination assay. The -317H1/H2 genotype was measured through PCR and restriction fragment length polymorphism analysis. Serum lipid and apo levels were measured by an enzymatic kit and a PEG-enhanced immunoturbidimetric assay. RESULTS: Allelic and genotypic frequencies of the APOC1 gene rs4420638A/G and -317H1/H2 were not significantly different between the two groups (all P> 0.05). However, patients carrying the G allele of the rs4420638A/G locus had higher serum levels of triglyceride, non-HDL-C and apoB, and a higher apoB/apoA1 ratio compared with those with an AA genotype (all P< 0.05). Patients carrying the H2 allele of the -317H1/H2 polymorphism had smaller delivery gestational weeks compared with those with the H1H1 genotype (P< 0.05). CONCLUSION Polymorphisms of the APOC1 gene rs4420638 and -317H1/H2 sites may be associated with abnormal lipoprotein metabolism among Chinese patients with PE, though no association was found between variants of the APOC1 gene and the risk of PE among them.