Effects of liver regeneration on the proliferation of rat fetal hepatocytes after intrasplenical transplantation

AIM:To study the effect of environment of liver regeneration on the proliferation of rat fetal hepatocytes after intrasplenical transplantation. METHODS:Fetal hepatocytes isolated from 3-week SD rat fetuses bred were transplanted into the spleens of liver regeneration model rats with 70% partial hepatectomy. The cell cycle of the hepatocytes in the remnants liver was analyzed by flow cytometer and the density dimensions of the donor fetal hepatocytes in spleen were measured by image analysis system 7 and 30 days post-transplantation, respectively. RESULTS:Compared with the control group, the proportions of S and G₂ /M cells in the remnants liver were obviously decreased (P＜0.05), but the density dimensions of the donor fetal hepatocytes in spleen increased significantly (P＜0.05) in rats with hepatectomy 7 days post-transplantation. CONCLUSION:The environment of liver regeneration is propitious to the proliferation of fetal hepatocytes after transplantation into spleen.     

Se-containing spirulina phycocyanin attenuated liver injury induced by carbon tetrachloride in mice

AIM:To investigate the effect of Se-containing spirulina phycocyanin (Se-SPC) on liver injury of mice induced by carbon tetrachloride (CCl₄). METHODS:The mouse model was conducted by intragastric feeding with 2% CCl₄ oil for three times, meanwhile Se-SPC, spirulina phycocyanin (SPC) and Na₂SeO₃ were injected (ip) to various groups for 7 days. Then selenium (Se), glutathione peroxidase (GPx), superoxide dismutase (SOD), alanine aminotransferase (ALT), malondiaoldehyde (MDA) and nitric oxide (NO) levels in blood and liver were measured. RESULTS:The level of Se,GPx and SOD activities were obviously higher(P<0.05)but ALT activity,MDA and NO₂^-/NO₃^- levels were remarkably lower(P<0.05)in Se-SPC treated groups than those in CCl₄ groups,and effects of high dose Se-SPC on Se,GPx,MDA and NO₂^-/NO₃^- were even more significant(P<0.01).Under the same dose of Se or protein,effects of all selected targets in Se-SPC groups were more efficient than those in SPC groups and inorganic-Se groups.Furthermore,Se levels had a positive correlation with GPx activity(r=01705),which had negative correlation with levels of MDA,NO₂^-/NO₃^- and ALT(r=-0.629,r=-0.336,r=-0.457,respectively), and positive correlations between ALT activity and MDA or NO₂^-/NO₃^- level were found (r=0.519,r=0.641). CONCLUSION:These results indicated that Se-SPC may attenuate liver injury of mice induced by CCl₄ through its anti-inflammatory action and enhancing selenoenzyme expression.     

Effects of nitric oxide on hepatic encephalopathy in cirrhotic rats induced by LPS

AIM: To investigate the effects of nitric oxide (NO) on hepatic encephalopathy in cirrhotic rats induced by LPS. METHODS: The cirrhotic model of rats was established by complex pathogeny. Since the end of the 8 th week, the rats were intragastrically-infused with 0.9% salt, L-arginine(L-arg) and LNNA respectively for 2 weeks.The hepatic encephalopathy in cirrhotic rats were induced by 3 mg/kg LPS (ip) 4 hours before the rats were sacrificed. RESULTS: The normal behaviors and electroencephalograph were appeared in L-arg group. LNNA group showed hepatic encephalopathy. The content of NO₂^-/NO₃^- of brain tissue was markedly higher in L-arg group than LNNA group(P<0.05), but the content of histamine in brain tissue was lower in L-arg group than LNNA group(P<0.05). There was a negative correlation between the content of histamine in brain tissue and the content of NO₂^-/NO₃^- of brain tissue. CONCLUSION: NO can prevent hepatic encephalopathy in cirrhotic rats induced by LPS.     

Expression of undulin in experimental rat liver fibrosis

AIM: To observe the expression of undulin (Un) in liver tissue and to clarify the diagnostic significance of serum Un in experimental rat liver fibrosis. METHODS: Rat liver fibrosis was induced by carbon tetrachloride. The expression of Un in liver tissue was detected by immunohistochemical method. Serum Un levels was measured by enzyme immunoassay. RESULTS: Expression of Un increased in fibrotic liver than normal liver, and it was mainly distributed in portal tract stroma, central veins and fibrotic septa in fibrotic liver. Also, the level of serum Un was significantly higher in fibrotic liver than normal liver. CONCLUSION: These data suggest that Un should be a component of the hepatic extracellar matrix, and its expression could be increased greatly in fibrotic rat liver. Serum Un levels may be used as an indicator in liver fibrosis diagnosis.     

高效液相色谱法测定氟甲喹可溶性粉含量

采用高效液相色谱法测定氟甲喹可溶性粉的含量,采用C 18 色谱柱(150 mm×4.6 mm,5μm),以乙腈-水-磷酸(35∶65∶0.5,V/V)为流动相,柱温25 ℃,流速1.0 mL/min,检测波长248 nm.方法的线性范围 100～400 μg/mL,线性方程A=2.455×104C 28 111,r=0.999 9,平均回收率100.3%,RSD= 0.74%.本法具有专属性,可将氟甲喹与已知的杂质进行有效分离.     

Model establishment and biological behaviour observation of mouse blastocyst co-cultured with hepatocarcinoma cell lines with differently invasive and metastatic potential in vitro

AIM: To explore interaction and biological behaviour changes of two kinds of cells-blastocysts and hepatocarcinoma cells in the same microenvironment. METHODS:The models of mouse blastocysts co-cultured with human hepatocarcinoma cell lines were established, then biological behaviours and mutual effects of the two kinds of cells in co-culture system were observed. RESULTS: Compared with control group, hepatocarcinoma cells with differently invasive and metastatic potential significantly enhanced the rates of blastocyst hatchment , attachment and outgrowth(P＜0.05). There was no significant difference in those among hepatocarcinoma cells co-cultured groups (P＞0.05). The blastocyst hatched and attached to hepatocarcinoma cells with differently invasive and metastatic potential. Then, differential trophoblasts invaded hepatocarcinoma cells. The clear-cut interfaces were gradually formed between both sides. Hepatocarcinoma cells on interface showed changes of growth direction and cell shapes and did not invade blastocysts. CONCLUSIONS: Hepatocarcinoma cells promoted blastocyst development. Blastocysts implanted and invaded hepatocarcinoma cells with differently invasive and metastatic potential in vitro, which indicate that blastocyst implantation in vitro does not relate with the kinds and differential level of interactional cells and the low selectivity maybe relate with high adaptability of early life.     

Characteristics in the secretion of liver TNF-α induced by infusion of LPS into veins in goats

AIM: To study the role of liver in immune regulation in experimental endotoxemia. METHODS: 17 castrated male goats were subjected to simultaneously installing catheters in jugular, hepatic and portal veins by surgery. Four days later, lipopolysaccharide (LPS) was infused in term of three groups as followings: In group ①, LPS of 20 EU (endotoxin unit, EU)·kg^-1 was infused into portal vein; In group ②, LPS of 20 EU·kg^-1 was infused into jugular vein and LPS of 1 500 EU·kg^-1 infused into jugular vein in group ③. Before and after infusion, blood samples were collected from the three veins through the catheters for 8 h.The plasma levels of TNF-α were measured by RIA. RESULTS: In group ①, the plasma TNF-α levels of hepatic and portal vein rose to peak value at 5 h, but that of the jugular vein did not changed. In group ②, the plasma TNF-α levels in hepatic vein rose to peak value at 3 h. The TNF-α levels of jugular vein rose to peak value at 1 h and the one in portal vein enhanced continuously between 0-8 h. In group ③, the plasma TNF-α levels in jugular, hepatic and portal vein rose to significant peaks at 1 h simultaneously. CONCLUSION: During experimental endotoxemia,liver showed different dynamic characteristics in TNF-α secretion according to the pathway and doses of LPS delivery.     

Effects of changing PKC activity on proliferation and telomerase expression in human hepatocellular carcinoma cells

AIM: To investigate whether protein kinase C (PKC) is involved in the proliferation and the telomerase expression in human hepatocellular carcinoma cells. METHODS: Human hepatocellular carcinoma cells (BEL-7402) were treated with exogenous phorbol-12-myristate-13-acetate (PMA, PKC activator) and staurosporine (SP, PKC inhibitor) for 48 hours. The techniques of cell culture and the telomeric repeat amplification protocol silver staining in combination with computer image scanning system in vitro were used to observe the variations of the growth and the telomerase expression. RESULTS: The proliferative potential of BEL-7402 cells was decreased by the action of PMA as well as SP, and the telomerase expression was also inhibited by PMA and SP. CONCLUSION: Our findings suggest that the proliferation of human hepatocellular carcinoma cells and the telomerase expression may be related to PKC.     

Effects of inhibition of Kupffer cell and splenectomy on thioacetamide-induced hepatic injury

AIM: To examine the effects of inhibition of Kupffer cell and splenectomy on intestinal endotoxemia and hepatic injury. METHODS: The hepatic injury model was established by treatment with thioacetamide (TAA). At the same time, inhibition of Kupffer cells by intravenous GdCl₃ and splenectomy were performed. Serum alanine aminotransferase (ALT), TNF-α, endotoxin content and phagocytic index were observed. RESULTS: In the TAA+GdCl₃ group, and TAA+splenectomy group, the endotoxin content was significently higher than that in normal and TAA group (P<0.05). The plasma TNF-α, ALT and total bilirubin were significantly lower than those in TAA group (P<0.05). CONCLUSION: Inhibition of Kupffer cells and splenectomy increase plasma endotoxin level, but decreases the plasma TNF-α levels and alleviates the hepatic injury induced by TAA.     

UHPLC-Q/TOF MS结合HPLC-DAD定性和定量分析硫酸黏菌素可溶性粉中的未知添加物

定性和定量分析一批兽药硫酸黏菌素可溶性粉中的未知添加物。照《中国兽药典》2010年版一部对该批检品用微生物检定法进行含量测定时,发现该样品的抑菌圈为虚圈,用薄层色谱鉴别该样品,未显示与标准品溶液一致的主斑点,怀疑该样品中有处方外非法添加物。采用超高效液相色谱-四级杆-飞行时间质谱(UHPLC-Q/TOF MS)对该样品进行筛查,发现疑似添加物,并使用液相色谱-二极管阵列检测(HPLC-DAD)法进行了双重确证和含量测定。该样品中非法添加物确证为磺胺氯达嗪和甲氧苄啶,添加量分别为58.5 mg/g和13.4 mg/g。本研究通过建立筛查方法为监管部门提供技术支撑,通过分析非法添加物的可能原因为打击兽药处方外非法添加提供了思路。