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 therapeutic effect of Chinese herbal medicines, which have the actions of dispersing stagnated liver-Qi and promoting bile secretion, clearing heat and removing damp, activating blood and removing blood-stasis, in the treatment of the third-level or above intrahepatic biliary duct stone. [Methods] After the routine surgical treatment including cholecystectomy, choledochotomy with T-tube drainage and choledochoscope lithotomy, 42 cases of third-level or above intrahepatic biliary duct stone received Dandao Paishi Mixture (DPM, mainly composed of Herba Lysimachiae, Radix Clematidis, Herba Artemisiae Scopariae, Radix Aucklandiae, Rhizoma Curcumae Longae, Radix Curcumae, etc.) for oral use. The effects of DPM on the intrahepatic biliary duct stone were evaluated after treatment. [Results] Before the treatment of DPM, the stones in third-level or above intrahepatic biliary ducts cannot be taken out; after the treatment of DPM, T-tube reverse biliary contrast examination showed that stones were removed in 39 (92.9%) cases, stones still existed in the common bile duct in 3 cases and then were removed by lithotomy through T-tube sinus of the common bile duct with Olympus fiber choledochoscope 8 weeks later. [Conclusion] DPM has good lytholytic and lithagogue effects for the treatment of third-level or above intrahepatic biliary duct stone and for intrahepatic biliary duct incarcerated stone. It can also reduce the postoperative residue of stone and recurrence rate.

Biomarkers ; Humans ; Idiopathic Pulmonary Fibrosis/genetics* ; MicroRNAs/metabolism* ; Sjogren's Syndrome/genetics*

ObjectiveTo retrospectively analyze the clinical outcome of patients with iatrogenic peripheral nerve injury so as to summarize the experiences and lessons.MethodsA retrospective study was performed on 72 patients with iatrogenic peripheral nerve injury treated in our hospital from 2004 to 2010.Therapy methods included conservative treatment in 24 patients,surgical release in 21 and nerve anastomosis in 27.Results All the patients were followed up for average 10 months ( range,3-24months).The neurological recovery was excellent in 24 patients,good in 21,fair in 16 and poor in 11,with excellent rate of 64％,according to the trial standard of upper limb functional assessment established by Chinese Medical Society of Hand Surgery.ConclusionsRisk awareness should be strengthened on the iatrogenic peripheral nerve injury,especially the minimally invasive treatment of fractures in recent years,which has increased the risk of nerve injury.For patients who may be presented with iatrogenic injury,we should develop a detailed preoperative surgical plan for early one stage treatment as possible.

Objective To summarize the clinical manifestation ,the main pathogenic bacteria distribution and drug resistance of neonatal community acquired sepsis late onset in our hospital .Methods Retrospectively analyse the clinical material of 122 cases (41 premature cases and 81 cases of full term) with neonatal community acquired sepsis late onset ,which were clinically diagnosed , from January 2009 to December 2012 in our hospital .Results The main clinical manifestation of neonatal community acquired sep-sis late onset was poor response(64 .7% ) ,repellent milk(57 .4% ) ,temperature changes(61 .5% ) ,and the respiratory tract and um-bilical region were the main infection ways .42 cases were checked out with pathogen in the 122 cases ,blood culture positive rate was 34 .4% ,and there was no statistically differences between the premature and the full term infant group ,In the 42 cases ,there were 29 cases with staphylococcus ,including 10 cases of staphylococcus aureus ,14 cases of coagulase negative staphylococcus and 5 cases of enterococcus ;and there were 10 cases are checked out with e .coli .All of the coccus detected were resistant to penicillin and erythromycin ,but sensitive to vancomycin ,teicoplanin ,linezolid .The e .coli was sensitive to amikacin ,imipenem ,meropenem ,and al-so had a higher sensitivity to cefazolin ,ceftriaxone ,cefepime ,cefoperazone and nitrofurantoin .Conclusion Blood culture positive rate is not high in neonatal community acquired sepsis late onset ,and its′clinical manifestations are nonspecific .The main pathogenic bacteria is coagulase negative staphylococcus ,staphylococcus aureus ,followed by escherichia coli .

Objective To investigate the incidence and possible reasons of patients with rheumatoid arthritis (RA) who stopped taking slow-acting antirheumatic drugs (SAADs).Methods Two hundreds and twenty-four in-or out-patients with RA were prospectively followed up for 4-year,and their demographic and clinical information and reasons of SAADs cessation were recorded.The difference between patients who stuck to SAADs and those who stopped SAADs and the factors that resulting the cessation were analyzed.Results ① Fourty-eight percent of 224 patients with RA ceased taking SAADs during 4-year period.② The main reasons of ceasing SAADs,provided by patients themselves,were side effects of SAADs,ineffectiveness,lack of knowledge about drugs,short of SAADs,concurrent diseases,and intention to pregnancy.③ There was significant difference in low income,poor educational status,number of tender joints,and rheumatoid factor level between the patients who stuck to SAADs and those who did not.Conclusion The high incidence of SAADs cessation in RA patients should rise concern.Both medical and patient factors contribute to the cessation.