Objective Human sperm hyaluronidase in the acrosome is a key enzyme during fertilization.To study and establish a modified substrate film method to improve the diagnotic and treatment of male infertility and investigate the influence of sperm hyaluronidase on male fertility.Methods According to the biochemical feature of sperm hyaluronidase that can dissolve the hyaluronic acid in matrixes,the modified sodium hyaluronate-Gelatin membrane was used as substrate to demonstrate the sperm hyaluronidase activity by incubation and staining.70 human semen samples were selected,categorized and detected for hyaluronidase activity according to the resuh of routine clinical semen examination.The average sperm hyaluronidase activity was statistically anyliezed between the fertile group and infertile group.Results Under general light microscope,the clear and bright digestion circle around the sperm head can be observed against the background of deep purple-blue stained substrate at the positive-reaction area.The amounts of positive reaction and the diameter of the bright circle are positively related to the activity of sperm hyaluronidase.The average hyaluronidase reaction rate and diameter of normal fertile group were 70.84% and 78.17 ?m;What about the infertile group A were 60.02% and 76.92 ?m;The infertile group B were 29.11% and 8.22 ?m; There was statistical difference of HYD activity between the infertile group.B and the fertile group(P

Animals ; Chromatography, Gas ; methods ; Hexanones ; blood ; Mice

Objective To evaluate left ventricular(LV) systolic function by two-dimensional speckle tracking imaging(2D-STI) in severe aortic stenosis(AS) patients.Methods Standard echocardiography and 2D-STI examinations were performed in a total of 54 subjects including 26 consecutive patients with severe AS with LV ejection fraction(LVEF) ≥50％ and 28 controls.2D-STI including systolic longitudinal strain (LS) and strain rate(LSr) were assessed from the apical 4-chamber,3-chamber and 2-chamber views,the circumferential strain(CS) and strain rate(CSr),radial strain(RS) and strain rate(RSr) were measured from the standard short axis views (averaging 6 segments per view).The above parameters of AS groups were compared with those of normals.The repeatability of LVEF,LS and RS was evaluated in 6 AS patients and 6 normal volunteers selected at random from the investigation.Results ① There was no significant difference between AS group and control group in LVEF,LV end-diastolic dimension(LVEDD),LV endsystolic dimension (LVESD) and midwall fractional shortening (mwFS) (P ＞ 0.05).② Significant differences were detected between the two groups.LS,RS,CS,LSr,RSr,and CSr were obviously decreased in AS group(P ＜0.05).③The repeatability of LVEF,LS and RS was good by consecutive measurement of identical and independent observers.Conclusions Despite the presence of normal LVEF,the LV systolic function is impaired in severe AS patients.

Objective To determine the prevalence and genotype profiles of urogenital Chlamydia trachomatis and their related factors in patients attending sexually transmitted diseases (STD) clinics in Guangxi Zhuang Autonomous Region. Methods C. trachomatis was screened by a plasmid PCR in 598 patients attending STD clinics. Then, positive specimens underwent nested-PCR to amplify the major outer membrane protein 1 (ompl) gene. The amplicons of ompl gene were digested by restriction endonucleases Alu I plus Hinf I and Cfol . C. trachomatis was differentiated according to the restriction fragment length polymorphism (RFLP) patterns. Results Out of the 598 samples, 83 were positive for plasmid-PCR. The prevalence of chlamydial infection was 13.9% with no significant difference between male and female patients. Nested-PCR based RFLP analysis showed that genotype E amounted to 27.7% (23/83), F 20.5% (17/83), D 13.2%(11/83), G 12.0%(10/83), K 7.2%(6/83), H 4.8%(4/83), I 3.6% (3/83), J 3.6%(3/83)and uncertain types 7.2% (6/83). Visible symptoms were observed less frequently in infections with C. trachomatis genotypes E and F compared with the other genotypes, while low abdominal pain occurred in 80% of infections with type G. Conclusions A certain proportion of out-patients attending STD clinic are infected with various types of C. trachomatis in Guangxi Zhuang Autonomous Region. The polymorphism of ompl gene may serve as a useful tool in molecular epidemiological studies of C. trachomatis.

ObjectiveTo define the maximum tolerated dose (MTD) of weekly cisplatin in concurrent chemoradiotherapy for Chinese cervical carcinoma.MethodsCervical carcinoma of stage ⅠB2- ⅣA were eligible for the study.PhaseⅠstudy was dose-escalation trial with 15 patients.All patients received whole pelvic radiotherapy with three dimentional conformal radiotherapy technique. Concurrent cisplatin started from the dose of 20 mg/m2 to 25 mg/m2,30 mg/m2,35 mg/m2,40 mg/m2 for the weekly schedule ( ≥3 patients per dose group) and the doses were steadily escalated to 40 mg/.m2.If the dose was increased to 40 mg/m2 without dose-limiting toxicity ( DLT),40 mg/m2 would be the maximum tolerated dose (MTD).According to the MTD dose from Phase Ⅰ study,we conducted phase Ⅱ clinical trial with 36 patients.ResultsIn Phase Ⅰ study,cisplatin dose was escalated to 40 mg/m2 and DLT had not been reached.Thirty-six patients in Phase Ⅱ study included 9 inpatients and 27 outpatients.All 9 inpatients completed 6 cycles of chemotherapy. In 27 outpatients,18 patients (66％) completed 6 cycles of chemotherapy,19 patients (70％) completed 5 cycles and 25 patients (92％) completed 4 cycles of chemotherapy.All patients completed radiotherapy.Major adverse effects were grade 1 and 2 gastrointestinal toxicities and neutropenia.ConclusionsWeekly 40 mg/m2 cisplatin concurrent with radiotherapy is well tolerated when given to Chinese patients with cervical carcinoma. For outpatients with poor performance status,the cisplatin dose needs to be reduced.

Objective:To study the effects of Ophiocordyceps xuefengensis on proliferation of DC -CIK cells and the activity of killing HepG-2 cells by DC-CIK cells.Methods:Peripheral blood mononuclear cells were routinely isolated and induced into DC and CIK.DC and CIK co-cultured by 1∶5 for 7 days,then Ophiocordyceps xuefengensis were added into medicine group ,observed the mor-phology of the cells on the tenth day and counted the DC-CIK number of each group.DC-CIK cells acted as effector cells and the HepG-2 cells as target cells , cck-8 method for the detection of DC-CIK in the killing rate of HepG-2.Results: The Ophiocordyceps xuefengensis was able to proliferate the DC-CIK dramatically ,the optimal concentration was 0.1 mg/ml.Cultivation of Ophiocordyceps xuefengensis induced DC-CIK cells on HepG-2 cells killing effect was better than that of the routine method of DC-CIK cells; the effection of Ophiocordyceps xuefengensis killing HepG-2 cells was not obviously.Conclusion: Ophiocordyceps xuefengensis can enhance the anti-tumor activity of DC-CIK mainly by promoting the proliferation of it.

Objective To research mainly focuses on the changes of genomic DNA global methylation of peripheral blood CD14 + monocytes in patients with coronary artery disease (CAD).Methods CD14 + monocytes were isolated from peripheral blood of treatment group (CAD patients) and control group(no-CAD patients) by density gradient centrifugation and magnetic bead selection.We detected genomic DNA global methylation level with genome global methylation quantitation kit.Quantitive real-time polymerase chain reaction(qRT-PCR) was performed to detect the expression levels of DNA methyltransferases (DNMTs) and methyl binding protein 2 (MBD2) mRNA.Results In control group and treatment group,genomic DNA global methylation levels of CD14 + monocytes were 0.30 ± 0.04,0.44 ± 0.03,the difference was statistically significant(P ＜ 0.05).In control group and treatment group,the expression levels of DNMT1 mRNA were 2.16 ± 0.32,1.40 ± 0.13,with significant difference(P ＜0.05).The expression levels of DNMT3A mRNA were 0.56 ± 0.05,0.89 ±0.12,with significant difference(P ＜0.05).The expression levels of DNMT3B mRNA were 0.59 ±0.07,0.97 ±0.10,with significant difference(P ＜0.01).The expression levels of MBD2 mRNA were 1.68 ±0.20,1.20 ±0.08,with significant difference(P ＜0.01).In treatment group of coronary heart disease,global methylation level and the expression levels of DNMT1 mRNA (r =0.648) as well as DNMT3B (r =0.700) were positive correlated and statistically significant (both P ＜ 0.05),global methylation level and the expression levels of MBD2 (r =-0.540) were negative correlated and statistically significant(P ＜0.05),while global methylation level and the expression level of DNMT3A were not correlated,the difference was not significant (P ＞ 0.05).Conclusion Genomic DNA of peripheral CD14 + monocytes in patients with coronary artery disease presents global hypermethylation,which may be as a result of out-of-balance between methylation and demethylation induced by abnormal expressions of DNMTs and MBD2.

This article reviews the influencing factors of intestinal lavage in patients with defecation disorder after spinal cord injury, and summarizes the intervention methods to improve the effect of intestinal lavage, so as to provide a reference for the correct implementation of intestinal lavage and improve the quality of life of patients.

OBJECTIVETo explore the role of X-linked inhibitor of apoptosis protein (XIAP) in the fibronectin (Fn)-adhesion mediated drug resistance of HL-60 cells.

METHODSCulture plates were coated with Fn and bovine serum albumin (BSA) (as control), respectively. Colorimetric CCK-8 assay was used to determine the effects of Fn on the cytotoxicity of DNR to HL-60 cells. Intracellular DNR accumulation was assayed with flow cytometry. Reverse transcription-PCR and Western blot were used to examine the mRNA expression and XIAP, bcl-2, MRP and mdr1 proteins, respectively. HL-60 cells were added to Fn coated Culture plates. The fully phosphorothioate antisense oligonucleotide (AS-ODNs) and the control ODNs of XIAP were delivered into HL-60 cells in the form of liposome-ODN complexes. IC(50) was calculated by linear regression of survival percent versus drug concentration.

RESULTSHL-60 cells adhered to Fn-coated plates had a significant survival advantage over those grown on BSA coated plates and in suspension when exposed to DNR, the IC(50) of Fn group being significantly higher than that of BSA group and suspension group (0.526 micromol/L vs 0.132 micromol/L, 0.123 micromol/L, respectively, P < 0.05). XIAP was up-regulated significantly in Fn group compared with BSA group and suspension group (P < 0.05), whereas there was no difference in the expressions of bcl-2, MRP and mdr1 among the three groups (P > 0.05). The intracellular concentration of DNR in Fn-adhered HL-60 cells was similar to that in BSA group and suspension group (P < 0.05). AS-ODNs of XIAP down-regulated the XIAP expression in Fn-adhered HL-60 cells. In addition, AS-ODNs sensitized HL-60 cells to the cytotoxic effects of DNR.

CONCLUSIONThe increased XIAP protein level contributes to the drug resistance induced by adhesion to Fn. AS-ODNs of XIAP might reverse the drug resistance.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Cell Adhesion ; Daunorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; physiology ; Drug Resistance, Neoplasm ; genetics ; physiology ; Fibronectins ; HL-60 Cells ; Humans ; Multidrug Resistance-Associated Proteins ; genetics ; metabolism ; Oligonucleotides, Antisense ; genetics ; RNA, Messenger ; genetics ; Transfection ; X-Linked Inhibitor of Apoptosis Protein ; genetics ; metabolism ; physiology

OBJECTIVETo evaluate the effect of modified culture method used to cytogenetic analysis and the clinically significance of chromosomal abnormalities to multiple myeloma (MM).

METHODSMononuclear cells were isolated from bone marrow aspirate of 20 MM patients; and then cultured for 3 days without any cytokines, and 6 days in the presence of IL-6 (10 ng/mL) and GM-CSF (30 ng/mL) before RHG banding analysis; the remained part of aspirates were treated directly. Eight cases of iron deficiency anemia were taken as control.

RESULTSThe experiment was failure in 2 cases because of blood clot, and another 2 cases could be analyzed only by direct method due to inadequate cells. The karyotype abnormalities were found from 4 cases of 16 available patients. Of them, three cases had complex karyotypes. The abnormalities were detected after 6 days culture with addition of cytokines. No abnormalities were detected from those groups of directly analysis and 3 day culture. Meantime, the clinical data showed that the patients with cytogenetic abnormalities were in stage III, and had a high percentage of MM cells (25%-56%) in their bone marrow, and also poor responses to prior chemotherapy. No cytogenetic abnormalities were found from control individuals in all groups.

CONCLUSIONExtended culture in the presence of cytokines could improve the efficiency of cytogenetic analysis to MM. Complex karyotype was common cytogenetic abnormalities in MM patients with poor response to chemotherapy.

Aged ; Chromosome Aberrations ; Cytogenetic Analysis ; Cytokines ; metabolism ; Female ; Humans ; Karyotyping ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology