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ObjectiveTo study the effect of Qizhu Kang'ai prescription （QZAP） on the gluconeogenesis enzyme phosphoenolpyruvate carboxykinase 1 （PCK1） in the liver of mouse model of liver cancer induced by diethylnitrosamine （DEN） combined with carbon tetrachloride （CCl4） and Huh7 cells of human liver cancer， so as to explore the mechanism on regulating metabolic reprogramming and inhibiting cell proliferation of liver cancer cells. MethodDEN combined with CCl4 was used to construct a mouse model of liver cancer via intraperitoneal injection. A normal group， a model group， and a QZAP group were set up， in which QZAP （3.51 g·kg-1） or an equal volume of normal saline was administered daily by gavage， respectively. Serum and liver samples were collected after eight weeks of intervention. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， γ-glutamyltransferase （γ-GT）， and alpha-fetoprotein （AFP） in mice were detected to evaluate liver function changes of mice in each group. Hematoxylin-eosin （HE） staining and Sirius red staining were used to observe pathological changes in liver tissue. In the cell experiment， Huh7 cells were divided into blank group， QZAP low， medium， and high dose groups and/or PCK1 inhibitor （SKF-34288 hydrochloride） group， and Sorafenib group. The corresponding drug-containing serum and drug treatment were given， respectively. Cell counting kit-8 （CCK-8） method， colony formation experiment， Edu fluorescent labeling detection， intracellular adenosine triphosphate （ATP） content detection， and cell cycle flow cytometry detection were used to evaluate the proliferation ability， energy metabolism changes， and change in the cell cycle of Huh7 cells in each group. Western blot was used to detect the protein expression levels of PCK1， serine/threonine kinase （Akt）， phosphorylated Akt （p-Akt）， and cell cycle-dependent protein kinase inhibitor 1A （p21）. ResultCompared with the model group， the pathological changes such as cell atypia， necrosis， and collagen fiber deposition in liver cancer tissue of mice in the QZAP group were alleviated， and the number of liver tumors was reduced （P<0.01）. The serum ALT， AST， γ-GT， and AFP levels were reduced （P<0.01）. At the cell level， compared with the blank group， low， medium， and high-dose groups of QZAP-containing serum and the Sorafenib group could significantly reduce the survival rate of Huh7 cells （P<0.01） and the number of positive cells with Edu labeling （P<0.01） and inhibit clonal proliferation ability （P<0.01）. The QZAP groups could also reduce the intracellular ATP content （P<0.05） and increase the distribution ratio of the G0/G1 phase of the cell cycle （P<0.05） in a dose-dependent manner. Compared with the model group and blank group， PCK1 and p21 protein levels of mouse liver cancer tissue and Huh7 cells in the QZAP groups were significantly reduced （P<0.05，P<0.01）， and the p-Akt protein level was significantly increased （P<0.01）. Compared with the blank group， the ATP content and cell survival rate of Huh7 cells in the SKF-34288 hydrochloride group were significantly increased （P<0.05）， but there was no statistical difference in the ratio of Edu-positive cells and the proportion of G0/G1 phase distribution. Compared with the SKF-34288 hydrochloride group， the QZAP combined with the SKF-34288 hydrochloride group significantly reduced the ATP content， cell survival rate， and Edu-positive cell ratio of Huh7 cells （P<0.05） and significantly increased the G0/G1 phase distribution proportion （P<0.05）. ConclusionQZAP may induce the metabolic reprogramming of liver cancer cells by activating PCK1 to promote Akt/p21-mediated tumor suppression， thereby exerting an anti-hepatocellular carcinoma proliferation mechanism.
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Objective To explore the application value of detection of Mycoplasma,Chlamydia and anti-sperm antibodies for male infertility.Methods Culture method,immune chromatography,and enzyme linked immunosorbent assay were adopted for the detection of Mycoplasma,Chlamydia,and anti-sperm antibodies respectively in 102 cases of infertile males and 42 cases fertile males.And the routine semen analysis was proceed as well.All the subjects were divided into 4 groups according to the detection re-sults:simple Mycoplasma or/and Chlamydia positive group(71 cases),simple anti-sperm antibodies positive group(21 cases),My-coplasma and Chlamydia or/and anti-sperm antibodies positive group(8 cases),Mycoplasma,Chlamydia and anti-sperm antibodies negative group(44 cases).The main indexes of semen routine were compared among 4 groups.Results The positive rates of Myco-plasma,Chlamydia and anti-sperm antibodies in infertile males were significantly higher than those of fertile males (P <0.05).The sperm densities,activity rates,activity of simple Mycoplasma or/and Chlamydia positive group,simple anti-sperm antibodies posi-tive group,and Mycoplasma and Chlamydia or /and anti-sperm antibodies positive group were significantly lower than the nega-tive group,while the sperm malformation rates,liquefaction times,white blood cell counts of the three groups were significantly higher than the negative group(P <0.05 ).Conclusion Mycoplasma,Chlamydia infection and anti-sperm antibodies production have significant effect on the indexes of semen,which cause decline in semen quality and the occurrence of male infertility.
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Objective To explore the dynamic changes of von willebrand factor(VWF)and Pseleetin in the finger-replanted patients,and the relationship between the prognosis of the surgery and hypercoagulability.Methods From December 2004 to December 2006,eishty finger-replanted patients were recruited to our study.with 40 healthy volunteers as controls.Plasma VWF and P-selectin were detected by enzyme-linked immunosorbent assay(EUSA)in both controls and patients before or after replantation.Results The VWF and P-selectin levels had significant differences between the replantations and controls(F=14.76,11.76,P＜0.01).The VWF levels in the patients of 1,4,8,16 hours after replantation were(1 715±493),(1 396±549),(1 266±504),(1 163±436)U/L respectively,all markedly higher than the controls(P＜0.01).The P-selectin levels in patients of 1,4,8,16,24 hours after operation were(14.7±2.6),(12.5±3.0),(11.8±3.2),(11.1±3.0)、(10.5±2.6)μg/L,significanfly higher than the controls(P＜0.01).The VWF levels in patients of pre-replantion and the 1,4,8,16,24,48,72 hours after replantation were(854±209),(1 535±389),(1 177±407),(1 040±283),(958±216),(829±193),(777±151),(713±137)U/L in successful group,and were(1 202±164),(2 333±243),(2 146±161),(2 039±244),(1 865±170),(1 645±283),(1 427±331),(1 188±262)U/L in unsuccessful groups.They were all significantly different at the same test-time points between two groups(t=4.44,5.12,6.10,8.43,10.17,8.85,5.10.4.61,P＜0.05).The P-selectin levels in patients of 1,4,8,16,24,48,72 hours after replantation were(13.9±2.5),(11.2±2.0),(10.2±1.6),(9.6±1.2),(9.2±0.9),(9.5±0.6),(9.3±0.4)μg/L in successful group,and(17.2±1.0),(16.9±1.0),(17.0±1.3),(16.1±1.1),(14.9±1.5),(13.8±1.4),(12.8±1.2)μg/L in unsuccessful group.Significant difference existed at the same testtime points between two groups again(t=5.22.9.91,10.35,12.79,9.46.9.45,9.33,P＜0.01).After replantation,both VWF and P-selectin were rapidly elevated and went to the summit 4 hours later,then declined to pre-replantation level about 24 to 48 hours later after replantation.Conclusions VWF and P-selectin were associated with the hypercoagulability.Dynamic monitoring VWF and p-selectin may be useful in determining the existence of hypercoagulability and the therapy of anti-coagulability.