BACKGROUND: As a biochemical marker of cytoplasm, the increased activity of creatine kinase (CK) in human spermatozoa is correlated with both the residual cytoplasma and the ratio of sperm with abnormal func tion. It is a marker of mature sperm and associated with the potential of in semination. OBJECTIVE: To investigate the differences of CK activity in sperm and the relative contents of sperm CK-MM and CK-BB isoenzymes between normal fertile males and infertile males, and evaluate its significance in clinical diagnosis of male infertility. DESIGN: Case-control observation. SETTING: Laboratory of Biochemistry, Family Planning Research Insti tute of Sichuan Province. PARTICIPANTS: Ninety-four male infertile patients between January 1999 and October 2000 were selected from the Department of Family Planning Sciences of Sichuan Province, who had no aspermatism with their wives proved to be fertile. The average age of subjects were 31 years. Eighty subjects with the sperm count ＞ 2×1010 L-1 were taken as normal sperm group and 14 subjects with the sperm counts ＜ 2×1010 L-1 were considered as oligospermia group. Semen obtained from 27 healthy males who were normal in routine examinations and with children was taken as the healthy control group.METHODS: Semen sample collected by masturbation after abstinence of 3 to 5 days was incubated at 37 ℃ for liquefication and routinely analyzed.Total activity of CK in sperm was determined by using a kinetic spectrophotometry and the relative contents of CK isozyme was determined by agarose gel electrophoresis followed by density scanning of CK isozyme.MAIN OUTCOME MEASURES: Sperm counts, percentages of viability and motility of sperm, total CK activity and the relative contents of CK-MM and CK-BB isozyme in spermatozoa.RESULTS: A total of 94 enrolled patients and 27 normal controls were involved in the analysis of results. ①Sperm counts, percentage of viability and motility in oligospermia group ( Ⅱ + Ⅲ, WHO method) were obviously lower than those in the healthy control group, and those in the normal sperm group, except the sperm counts, were remarkably lower than the healthy control group [the sperm counts in healthy control group, normal sperm group and oligospermia group was (6.05 ±0.81 )×1010 L-1, (7.76±1.37)×1010 L-1 and (1.46±0.19)×1010 L-1 respectively (P ＜ 0.01). The survival rate in healthy control group, normal sperm group and oligospermia group was 85.1%,56.8% and 58.2%, P ＜ 0.01, and the sperm motility was 62.9% ,34.6% and 29.5% respectively, P ＜ 0.01].②Total activity of sperm CK in oligospermia group was significantly higher than the healthy control group [respectively (9.000±6.117) and (1.933 ±0.943) kat/108 sperm,P＜ 0.05],although that in the normal sperm group (2.800±0.862) kat/108 sperm was a little higher than the healthy control group, while there were no significant difference between the two groups (P ＞ 0.05).③The relative contents of sperm CK-MM isozyme in the normal sperm group and oligospermia group were obviously decreased (that in the healthy control group, normal sperm group and oligospermia group was 30.5%, 19.0% and 18.0% respectively,P ＜ 0.05), which implied a remarkable difference in sperm differentiation between healthy control group and the latter two groups.CONCLUSION: The determination of sperm CK is meaningful for the diagnosis and pathogenesis of oligospermia. More work should be done on the distribution of sperm CK-MM isozyme in different infertile population as well as its importance in the diagnosis of infertility.

Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Humans ; Kidney Diseases ; therapy ; Nephrotic Syndrome ; therapy ; Rituximab

[Objective]To evaluate the effect of the tissue engineered nerve for bridging and repairing verve defect.[Method]Human bone marrow stromal cells(hBMSCs) were purified with centrifugate method,cultured in DMEM,induced with ATRA,BDGF and affected by heregulin,forsholin,bFGF(basic fibroblast growth factor),and PDGF(platelet-derived growth factor) of hBMSCs).The protein positive rate of S100 and GFAP of hBMSCs were determined by immunohistochmical staining.The tissue engineered nerves were constructed with hBMSCs mixed with extra-cellular matrix(ECM) and polylactic acid(PLA) tube.A 10mm defect of sciatic nerve was created in 24 Wistar mouse right limbs and ramdonly divided into three groups: group A(n=8),nerve defects bridged with polylactic acid(PLA) tube containing induced Schwann cells mixed with ECM,group B(n=8),with PLA tube containing ECM,group C(n=8) with autologous nerve graft.Functional recovery of nerve was examined by electrophysiological method and histological changes were examined with histological stainning of nerve and measurement quantity of new axon.[Result]The Schwann cells were presented at 12 wks after operation.The histologic and functional recovery of nerves of group A and group C were better than those of group B.the showed significant difference between group A or group C and group B and no significant difference between group A and group C.(PAB=0.021,PBC=0.001,PAC=0.065).Degradation of PLA tubes showed in group A and group B.[Conclusion]Schwann cells could be induced from hBMSCs,and the tissue engineered nerves,which were contructed by induced Schwann cells mixed with ECM and PLA tube,could be used to bridged and repair the peripheral nerve defect.

In order to analyze the correlation between critical residues in the catalytic centre of BSH and the enzyme substrate specificity, seven mutants of Lactobacillus salivarius bile salt hydrolase (BSH1) were constructed by using the Escherichia coli pET-20b(+) gene expression system, rational design and site-directed mutagenesis. These BSH1 mutants exhibited different hydrolytic activities against various conjugated bile salts through substrate specificities comparison. Among the residues being tested, Cys2 and Thr264 were deduced as key sites for BSH1 to catalyze taurocholic acid and glycocholic acid, respectively. Moreover, Cys2 and Thr264 were important for keeping the catalytic activity of BSH1. The high conservative Cys2 was not the only active site, other mutant amino acid sites were possibly involved in substrate binding. These mutant residues might influence the space and shape of the substrate-binding pockets or the channel size for substrate passing through and entering active site of BSH1, thus, the hydrolytic activity of BSH1 was changed to different conjugated bile salt.
Amidohydrolases ; genetics ; metabolism ; Bile Acids and Salts ; metabolism ; Escherichia coli ; metabolism ; Gene Expression ; Lactobacillus ; enzymology ; genetics ; Substrate Specificity

Acute Disease ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Phytotherapy ; Stroke ; drug therapy

Objective To investigate the liver protective effects of Siphonostegia chinensis decoction in rats with liver fibrosis induced by carbon tetrachloride(CCl4).Methods A total of 120 male SD rats were randomly divided into six groups: a blank control group, a model group, a Siphonostegia chinensis decoction group, a Salvia miltiorrhiza decoction group, a Rhizoma polygoni cuspidati decoction group and a polyene phosphatidylcholine capsules group, with 20 rats in each group. The model of liver fibrosis in rats was induced by subcutaneous injection with CCl4, except the blank group. Except rats in the blank control group and the model group, the rats in the other four groups were treated with Siphonostegia chinensis decoction, Salvia miltiorrhiza decoction, Rhizoma polygoni cuspidati decoction and polyene phosphatidylcholine capsules(10 ml/kg). The serum level of tumor necrosis factor-α(TNF-α)was determined with enzyme-linked immunosorbent assay, the fibrosis stage assessment were determined by histopathological examination.Results Compared with the model group, Siphonostegia chinensis decoction group significantly reduced the stage of liver fibrosis(P<0.01)and there were no obvious difference in liver fibrosis stage among the Salvia miltiorrhiza decoction group, Rhizoma polygoni cuspidati decoction group and polyene phosphatidylcholine capsules group, and the Siphonostegia chinensis decoction group(allP>0.05).Compared with liver fibrosis model group(368.06±24.90)U/L, the serum level of TNF-α in the Siphonostegia chinensis decoction group(336.61± 20.20)U/L significantly reduced(P<0.01), and there were no obvious difference among the Siphonostegia chinensis decoction group, the Salvia miltiorrhiza decoction group(337.81±21.04)U/L, the Rhizoma polygoni cuspidati decoction group(338.95±24.43)U/L and the polyene phosphatidylcholine capsules group(337.11± 23.64) U/L(allP>0.05).Conclusion Siphonostegia chinensis decoction has certain liver protective effect in rats with liver fibrosis induced by CCl4.

A case of localized scleroderma associated with mandibular osteomyelitis is reported.The formation mechanism of the case is a-nalysised according to clinical characteristics and literature data.

Objective To explore the effects of donor dendritic cells(DC)treated with Ad-IL- 12p35siRNA on the survival of allogragft recipients.Methods The recombinant adenoviral vectors carrying IL-12p35siRNA and HKsiRNA were transfected into bone marrow derived DC of BALB/C murine.C57BL/6 recipients were infused with DG(Ad-IL-12p35siRNA DC,Ad-HKsiRNA DC and control DC)from BALB/C donors 7 days before cardiac allograft,the survival time of murine and the change of T_H 1 and T_H2(IL-2,IL-4,IL-10 and IFN-?)cytokine were observed.Results The survival time of p35 group(20.17?2.71)days was longer than that of control group(7.81?1.61)days and HK group(7.17?1.60)days.The concentration of IL-2 and IFN-?in p35 group were significantly lower than those of control group and HK group,otherwise were the concentration of IL-4 and IL-10. Conclusion Pretreatment of dondor dendritic ceils with Ad-IL-12p35siRNA could prolonged cardiac allograft survival in recipicents.

Objective:To investigate the expression levels of CXCR1,CXCR2 and their common ligand CXCL8 in peripheral blood mononuclear cells (PBMCs) and liver biopsy from the patients with hepatitis B related hepatocellular carcinoma and their clinical significances.Methods:Quantitative real time polymerase chain reaction (qRT-PCR) was used to detect the mRNA levels of CXCR1,CXCR2,CXCL8 in the peripheral blood mononuclear cells of thirty-six hepatitis B related hepatocellular carcinoma and the protein levels of CXCR1 and CXCR2 and CXCL8 in liver biopsy were detected by SP immunohistochemical method.The level of C-reactive protein in serum was determined by chemiluminescence immunoassay respectively.Then,the correlations between CRP and the mRNA of CXCR1,CXCR2 and CXCL8 were analyzed.Results:The mRNA levels of CXCR1 (0.952 7±0.197 2),CXCR2 (0.896 9±0.173 0),CXCL8 (1.771 9±0.248 9) in the PBMCs of hepatitis B related hepatocellular carcinoma were significantly higher than those in controls (P＜0.01).And the protein levels of CXCR1,CXCR2 and CXCL8 were also obviously increased in liver biopsy of hepatitis B related hepatocellular carcinoma (P＜0.05).In addition,there was positive correlations between the level of serum C-reactive protein and the mRNA expression of CXCR1 (r =0.54,P＜0.01),CXCR2 (r =0.49,P＜0.01),CXCL8 (r =0.63,P＜0.01).Conclusion:The levels of CXCR1,CXCR2 and CXCL8 significantly increased in hepatitis B related hepatocellular carcinoma patients and positively correlated with serum CRP,suggesting that CXCR1,CXCR2 and their common ligand CXCL8 signal transduction process may play a key role in the pathological process of hepatitis B related hepatocellular carcinoma,which may provide a new direction for the immune intervention therapy of hepatocellular carcinoma.

Objective To investigate the expression and clinical significance of CXC chemokine receptors 1 and 2 (CXCR1 and CXCR2) and CXCL8 in peripheral blood mononuclear cells (PBMCs) and liver biopsy tissues from patients with primary hepatocellular carcinoma (PHC).Methods Serum specimens were collected from 36 patients with PHC, 30 patients with liver cirrhosis and 28 healthy subjects.Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to measure the expression of CXCR1, CXCR2 and CXCL8 at mRNA level in PBMCs.Streptavidin-perosidase (SP) immunohistochemistry was used to detect the expression of CXCR1, CXCR2 and CXCL8 at protein level in liver biopsy tissues.Levels of C-reactive protein (CRP), alpha-fetoprotein (AFP) and ferritin (FER) in the serum specimens were detected by chemiluminescence immunoassay.Then the correlations between these markers were analyzed.Results All of the results showed that the expression of CXCR1, CXCR2 and CXCL8 at mRNA level in PBMCs from the PHC group were higher than those of the healthy control group (P<0.01) as well as those of the liver cirrhosis group (P<0.05).Up-regulated expression of CXCR1, CXCR2 and CXCL8 in patients with PHC were associated with the depth of tumor invasion, lymph node or distant metastasis, clinical stage and levels of CRP, AFP and FER in serum (P<0.05).The expression of CXCR1, CXCR2 and CXCL8 at protein level in liver biopsy tissues were also significantly increased in the PHC group in comparison with those of the healthy control group as indicated by the result of SP immunohistochemistry (P<0.05).Conclusion Levels of CXCR1, CXCR2 and CXCL8 in the patients with PHC are significantly increased and positively correlated with the levels of AFP, FER and CRP in serum, suggesting that the signal transduction process mediated by CXCR1, CXCR2 and their common ligand CXCL8 may play a key role in the pathological process of PHC.This study may provide a potential new strategy for immune intervention in hepatocellular cancer.