Objective To study the renal protective effects of superfine powder of Rhodiola rosea in diabetic nephropathy (DN)rats.Methods The DN rats model was established by one side nephrectomy and intraperitoneal injection of streptozotocin(STZ).Then rats were randomly divided into 6 groups:sham-operation group,model group,Rhodiola rosea common powder groups(3 g/kg·d-1),and superfine powder of low,medium and high dose groups (0.75、1.5、3.0 g/kg·d 1),10 rats in each group.Rats were given the relevant drugs respectively for 7 weeks.Finally the levels of fasting blood glucose (FBG),serum total cholesterol (STC),blood urea nitrogen (BUN),creatinine (Cr) and urinary protein (Upro) were detected.The kidney tissue was taken for histopathological observation.Results Compared with DN modelgroup [FBG,STC,BUN,Cr and Upro was (24.78±3.66)mmol/L,(4.07±1.32)mmol/L,(22.36±2.54)mmol/L,(86.78±7.47) μmol/L,(50.23±6.82)mg respectively],the rats in DN model group after treatment with common and superfine powder (medium and high dose groups) of Rhodiola rosea appeared different levels of improvement ofFBG [(18.67±2.74) mmol/L,(17.21 ±3.17)mmol/L,(15.48±2.46) mmol/L],STC [(3.33±0.87) mmol/L,(3.42 ± 0.74) mmol/L,(3.21 ±0.92)mmol/L],BUN [(15.43±2.04)mmol/L,(16.56±1.85)mmol/L,(12.44±1.36)mmol/L],Cr [(66.17 ± 4.43) μmol/L,(68.42 ± 5.06) μmol/L,(61.33±4.21)μmol/L] and Upro [(37.47±5.64) mg、(35.66 ± 4.72) mg,(27.37± 3.92) mg],P＜ 0.05.The pathologic changes of kidney tissue were improved,too.Conclusion Rhodiola rosea has a good renal protective effect in rats with DN.Furthermore,the superfine powder is more effective than the common powder in renal protective effects.

Objective To investigate the role of r-aminobutyric acid B receptor in the development of liver fibrosis.Methods Thirty-two Sprague-Dawley (SD) rats were divided into four groups including a control group,a model group,a baclofen group,and a CGP35348 group.Liver fibrosis was then induced by carbon tetrachloride (CCl4).Baclofen and CGP35348 treatment were carried out after the formation of liver fibrosis,followed by complete extraction of the eyeball to obtain blood sample to test liver function.Liver tissue specimens were cut and stored for histological staining,histochemistry,real-time polymerase chain-reaction (RT-PCR),and western blot analysis.Results Histological staining indicated that the degree of liver fibrosis was more severe in the CGP35348 group than in the baclofen group (P＜0.001).The levels of alanine transaminase (ALT),aspartate aminotransferase (AST),gamma-glutamyl transferase (GGT),total bilirubin (TBil),and direct bilirubin (DBil) were significantly lower in the baclofen group than in the CGP35348 group (P＜0.01).The levels of ALT,AST,GGT,TBil,and DBil were significantly higher in the CGP35348 group than in the model group (P＜0.05).Immunofluorescence results show that the hepatic cell migration was inhibited in the baclofen group.Western blot results showed that the expression levels of α-SMA protein were significantly lowered in the baclofen group when compared to that of the CGP35348 group and model group (P＜0.01).Conclusion GABAB receptor might play a role in the liver protection by inhibition of migration of hepatic cells in liver fibrosis.Further studies into the mechanism behind this function are further needed and may be a potential source of future anti-fibrotic treatment.

AIM: To investigate the effects of octreotide (Oct) on the proliferation and extracellular matrix (ECM) synthesis in hepatic stellate cells (HSCs). METHODS: HSCs were isolated from normal male Sprague-Dawley rat liver by a combination of pronase-collagenase perfusion and density gradient centrifugation. The concentration of 2.5 ?g/L transforming growth factor ?1 (TGF?1) was used in all the experiment settings. Oct at concentrations of 0.01 mg/L ,0.1 mg/L,1 mg/L and 10 mg/L,respectively,or 0.01 mg/L Oct + TGF?1,0.1 mg/L Oct+TGF?1,1 mg/L Oct+TGF?1,10 mg/L Oct+TGF?1 were respectively added to the cultured HSCs. Effects of Oct on HSC proliferation and ECM synthesis were respectively determined by MTT method,-TdR and -proline incorporation,or radioimmunoassay. RESULTS: Oct inhibited MTT intake by cultured hepatic stellate cells and down-regulated -TdR incorporation,compared with control group. The concentrations of hyaluronic acid,laminin,collagen type IV in the culture supernatant and -proline incorporation in HSCs were decreased by Oct. TGF?1 obviously up-regulated proliferation and ECM synthesis in cultured HSCs,and Oct significantly blocked these actions. CONCLUSION: Oct inhibited proliferation and ECM synthesis in cultured HSCs,and elicited the effects of anti-hepatofibrogenesis.

Objective To summarize clinical outcomes of interventional therapy on children with common congenital heart diseases(CHD).Methods A retrospective study was conducted.One hundred and fourteen patients with CHD were selected as our subjects,who underwent catheter interventional therapy in the Second Affiliated Hospital of Zhengzhou University from Jan.2004 and Dec.2012.The size of occluder was chose according to intraoperative echocardiography or cardiac imaging measurements,and occluder was released under assisted monitoring by subtraction angiography or cardiac ultrasound.Results There are all together 112 patients got the therapy successfully,2 cases failed(occluder detachment),and the success rate of operation was 98.2％.After the success of interventional,echocardiography examination showed that 11 cases were with star point across shunt,but the function of the around valve was not affected.One months after operation,echocardiography examination showed star point across shunt of 11 cases were disappeared,and no occluder was shifted as well as no thrombosis formed.Three months after operation,chest radiograph showed pulmonary congestion decreases and heart shadow was shrink.Thirty-eight cases were with three tricuspid regurgitation before operation and 32 cases were without reflux at 3 months after operation,and 6 cases relieved significantly.The patients were followed up for 6 months or 3 years,activity endurance was significantly improved than that before operation.No occluder was shiftand hemolysis and arrhythmia occurred.Meanwhile,No thrombosis or embolism occurred.Conclusion Interventional treatment for children with congenital heart disease is proved as a safe,effective methods and it have broad prospects in clinical application.

Objectives　To assess the effect of an ACE inhibitor and an Ang Ⅱ type 1 (AT1) receptor antagonist on preventing hepatic fibrosis induced by CCl4 in rats and to investigate whether there is the expression of AT1 receptors on hepatic stellate cells. Methods　Studies were conducted in male Sprague-Dawley rats. Except for model group and control group, in three treated groups, either enalapril (5?mg/kg), or losartan (10?mg/kg), or enalapril+losartan were given to the fibrotic rats (daily gavage). Saline vehicle was given to the control group. After 6 weeks, liver fibrosis was assessed directly by hepatic morphometric analysis. The expression of AT1 receptors and α-smooth muscle actin (α-SMA) in liver tissue and isolated hepatic stellate cells (HSC) were detected by immunohistochemical techniques. Results　Compared with the fibrosis in rats of the model group, rats treated with either enalapril or losartan, or a combination of two drugs, showed a limited expansion of the interstitium (P<0.05), but no significant difference was observed among the three treated groups (P>0.05). The expression of AT1 receptors was found in abundance in the fibrotic interstitium of the fibrotic rats, whereas in the normal control rats they were limited to the vascular wall. AT1 receptors were also expressed on activated HSC in culture plates. Conclusions　Angiotensin-converting enzyme inhibitors and AT1 blockers might slow the progression of hepatic fibrosis. Activated HSCs expressed AT1 receptors. Activation of RAS might be related to hepatic fibrogenesis induced by CCl4.

OBJECTIVETo examine the expression changes of activin beta A, beta C, beta E and follistatin mRNA in the development of rat hepatic fibrosis induced by carbon tetrachloride (CCl(4)).

METHODSHepatic fibrosis was induced in rats by subcutaneous injections of 40% carbon tetrachloride oily solution for a period of 1 to 7 weeks. After carbon tetrachloride injection of 1, 2, 3, 4, 5, 6, and 7 weeks, the 6-12 rats were killed every time. The kinetics of activin beta A, beta C, beta E and follistatin mRNA expression were assessed by the semi-quantity RT-PCR.

RESULTSActivin beta A, beta C, beta E and follistatin mRNA could be detected in normal rat livers. After CCl(4) injection for 2 or 3 weeks, beta A mRNA was transiently decreased and became undetectable, then increased gradually. After CCl injection for 6 and 7 weeks, beta A mRNA level was significantly higher than controls (P<0.01). beta C mRNA could be detected after CCl(4) injection for 1 to 4 weeks and was significantly increased after 5 weeks over controls (P<0.05). beta E mRNA could not be detected after CCl(4) injection for 1 to 5 weeks, but significantly increased after CCl(4) injection for 6 or 7 weeks compared with controls (P<0.01). Except for normal rat liver, no follistatin mRNA was detected in rats after CCl(4) injection.

CONCLUSIONSActivins and follistatin have different expression changes in the development of hepatic fibrosis and the imbalance of activins and follistatin expression may involve in the formation of hepatic fibrosis.

Activins ; genetics ; Animals ; Carbon Tetrachloride ; Follistatin ; Gene Expression ; Inhibin-beta Subunits ; genetics ; Liver Cirrhosis, Experimental ; chemically induced ; genetics ; pathology ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

Purpose: C-X-C motif chemokine receptor 4 (CXCR4) and integrin αvβ6 play important roles in the malignant progression of multiple cancers. However, it remains unclear whether the expression of one or both proteins in breast cancer (BC) is of clinical significance. In this study, we investigated the expression of CXCR4 and integrin αvβ6 in BC tissues and their correlation with clinicopathological characteristics, including survival. Methods: CXCR4 and αvβ6 expression in 111 BC tissues was examined by immunocytochemistry. Correlations between the expression of the 2 proteins and patient clinicopathological characteristic were investigated using the Kaplan–Meier method and the Cox proportional hazards model. Results: CXCR4 and αvβ6 were overexpressed in BC tissue compared with normal breast tissue. Overexpression of both molecules was related to lymph node status (p = 0.013 and p = 0.022, respectively). αvβ6 overexpression was also associated with tumor size (p = 0.044). A positive correlation was detected between the expression of CXCR4 and αvβ6 (r = 0.649, p = 0.001), and co-overexpression of both molecules was associated with tumor size (p = 0.018) and lymph node metastasis (p = 0.015). Kaplan–Meier analysis revealed that overexpression of CXCR4, αvβ6, or both molecules was associated with short overall survival (OS; p < 0.001, p < 0.001, and p = 0.009, respectively) and disease-free survival (DFS; p < 0.001, p = 0.005, and p = 0.019, respectively). Multivariate analysis indicated that lymph node metastasis was an independent prognostic factor for unfavorable OS and DFS (p = 0.002 and p = 0.005, respectively), whereas co-overexpression of CXCR4 and αvβ6 was an independent prognostic factor only for OS (p = 0.043). Conclusion CXCR4 and αvβ6 may play synergistic roles in the progression of BC, and co-targeting of CXCR4 and αvβ6 could be a potential strategy for the prevention and treatment of BC.

Objective To compare the advantages and disadvantages of the three isolation regents for isolating primary hepatic stellate cells(HSCs) of mice.Methods The mice HSCs was isolated by in situ perfusion of per-perfusion solution,perfusion with collagenase Ⅳ.After digestion,Ficoll-Paque PLUS,perco11 and Nycodenz were use for the gradient centrifugation.To calculate the quantity,purity and the viability of the HSCs.The autofluorescence and Oil O staining were used to identify primary HSCs.Results The quantity of HSCs was (1.0 ± 0.5) × 106,(1.0 ± 0.5) × 105,(0.8 ± 0.5) × 106/ g_liver,respectively.Using Ficoll-Paque PLUS,percoll and Nycodenz,the purity of primary HSCs were 75％-85％,91％-95％,91％-95％,respectively.The viability were all above 90％.Conclusion compared with Ficoll-Paque PLUS and percoll,the Nycodenz is the the best separating medium for isolation and purification of mice primary HSCs.

Objective To evaluate the role of serum GP73 for diagnosing decompensated cirrhosis in population with chronic HBV infections.Methods The present study included three populations:200 patients with chronic HBV infections and receiving liver biopsy; 200 patients with decompensated cirrhosis;and 200 patients with hepatocellular carcinoma.All patients were HBsAg-positive over six months.Results Comparing with those in the patients with chronic hepatitis B (67.38 ±57.45 ng/ml),the serum GP73 levels in cirrhosis patients (221.9 ± 108.5 ng/ml) and HCC patients (152.4 ± 102.7 ng/ml) were significantly increased.Taken the population with chronic hepatitis B as "control group",the patients with decompensated cirrhosis as "patients group",the area of ROC analysis was 0.91 (95％ CI:0.88-0.94) (P ＜0.0001).Set the cut-off value at 150 ng/ml,the diagnosing sensitivity and specificity were 72.5％ and 93.5％,respectively.Conclusion For patients with chronic HBV infections and a higher GP73 levels,decompensated cirrhosis should be considered,except diagnosis of hepatocellular carcinoma.

Objective To evaluate the serum levels of GP73 for diagnosing autoimmune liver disease.Methods Three populations were included:80 patients with autoimmune liver disease; 80 patients with chronic hepatitis B,and 80 apparently healthy controls.Results The serum levels of GP73 in patients with autoimmune liver disease (112.3 ± 72.55 ng/ml) were significantly higher than those of patients with chronic hepatitis B (86.44 ± 60.69 ng/ml),and healthy controls (35.84 ± 11.8 ng/ml).However,there were no significant differences was observed in group of subjects with autoimmune hepatitis (107.4 ± 90.6 ng/ml),primary biliary cirrhosis ((89.0 ± 45.38 ng/ml),and primary sclerosing cholangitis (113.3 ± 50.87 ng/ml).Chosen the healthy control as "control group",the patients with chronic hepatitis B as "patients group ",the sensitivity and specificity of GP73 for diagnosing autoimmune liver disease were 75.0％ and 97.53％,respectively.The area of ROC analysis was 0.93(95％ CI;0.89-0.97).Conclusion The serum GP73 levels was significantly increased in patients with autoimmune liver disease,compared with those of healthy controls.For patients with higher levels of serum GP73,diagnosis of autoimmune liver disease should also be considered,especially for those patients with overlap-syndrome,except for chronic virus hepatitis and hepatocellular carcinoma.