2.Research of the improved fixed-substrate film for detecting of sperm acrosome hyaluronidase
Ke WANG ; Xiao-Qin GAO ; Xian-Lin LIU ;
Chinese Journal of Laboratory Medicine 2003;0(08):-
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
3.Evaluation of left ventricular myocardial function in patients with severe aortic stenosis by two-dimensional speckle tracking imaging
Hanjing GAO ; Mingxing XIE ; Li YANG ; Xiaojuan QIN ; Lin HE ; Xiaofang LU ; Ke LI
Chinese Journal of Ultrasonography 2013;22(11):921-926
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.
4.Molecular epidemiology of urogenital Chlamydia trachomatis infection in patients attending sexually transmitted diseases clinics
Bangyong ZHU ; Yongzhen LI ; Xing GAO ; Jie ZHANG ; Geng HUANG ; Shanlie QIN ; Xiumei ZHAO ; Yuqing WU ; Qiu KE ; Quan GAN
Chinese Journal of Dermatology 2008;41(5):282-284
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.
5.A phase Ⅰ / Ⅱ study of concurrent cisplatin chemotherapy in patients with carcinoma of the cervix receiving pelvic radiotherapy
Xiaomei LI ; Qingguo WANG ; Fuquan ZHANG ; Ke HU ; Min ZHANG ; Hongzhen LI ; Qing ZHAO ; Shangbin QIN ; Dongliang HOU ; Xianshu GAO
Chinese Journal of Radiation Oncology 2012;21(1):52-55
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.
6.Effects of Ophiocordyceps xuefengensis on proliferation of DC-CIK cells and activity of killing HepG-2 cells by DC-CIK cells
Bing ZHENG ; Fangyi XIE ; Guohui CAI ; Rucai ZHU ; Ke LI ; Shouquan GAO ; Dianbo TAN ; Xiaoyong HAO ; Yuhui QIN
Chinese Journal of Immunology 2015;(2):189-192
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.
7.Research of aberrant global DNA methylation level in CD14+ monocytes in patients with coronary artery disease and its mechanism
The Chinese Journal of Clinical Pharmacology 2017;33(18):1740-1743
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.
8.Research progress on influencing factors of intestinal lavage in patients with defecation dysfunction after spinal cord injury
Lifeng YAO ; Qin JIA ; Qin ZHOU ; Jun LI ; Shanshan CHEN ; Ping GAO ; Ke LI
Chinese Journal of Modern Nursing 2022;28(1):116-120
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.
9.Effect of 2,5-hexanedione on light-molecular-weight neurofilaments (NF-L) degradation of rat nerve tissues.
Chao-shuang ZOU ; Ke-qin XIE ; Rui-rui KOU ; Yuan GAO ; Fu-yong SONG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2012;30(6):418-422
OBJECTIVETo investigate the effect of 2,5-hexanedione (HD) on degradation of low-molecular-weight neurofilaments (NF-L) in nervous tissue of rats, and to explore the molecular mechanism of n-hexane neuropathy.
METHODSFifty male Wistar rats were randomly divided into one-week poisoning group (n = 10), two-week poisoning group (n = 10), three-week poisoning group (n = 10), four-week poisoning group (n = 10), and control group (n = 10). In the four poisoning groups, a rat model of n-hexane neuropathy was established by intraperitoneal injection of HD (400 mg/kg/d). The change in the sciatic nerve ultrastructure of each rat was observed under an electron microscope. The progression of HD-induced peripheral neuropathy was evaluated using a gait scoring system. The degradation rates of NF-L in the sciatic nerve and spinal cord of each rat were measured by Western Blotting.
RESULTSThe rats showed decrease in muscle strength and abnormal gait after two weeks of HD poisoning and mild or moderate paralysis after four weeks of HD poisoning. The sciatic nerve showed degenerative change, according to electron microscope observation. Compared with the control group, the two-week poisoning group, three-week poisoning group, and four-week poisoning group had the NF-L degradation rates decreased by 25.8%, 70.4%, and 69.7%, respectively, in the supernatant fraction of sciatic nerve, and by 14.7%, 64.6%, and 67.3%, respectively, in the sediment fraction of sciatic nerve, all showing a significant difference (P < 0.01). Compared with the control group, the one-week poisoning group had the NF-L degradation rate decreased by 33.87% in the supernatant fraction of spinal cord, the four-week poisoning group had the NF-L degradation rate increased by 16.2% in the supernatant fraction of spinal cord, and the one-week poisoning group and two-week poisoning group had the NF-L degradation rates decreased by 46.3% and 13.0% in the sediment fraction of spinal cord, all showing a significant difference (P < 0.01).
CONCLUSIONHD poisoning significantly inhibits NF-L degradation in the sciatic nerve, which may be associated with NF degeneration and accumulation in the axons of patients with n-hexane neuropathy.
Animals ; Hexanes ; poisoning ; Hexanones ; pharmacology ; Male ; Nerve Tissue ; drug effects ; metabolism ; physiopathology ; Neurofilament Proteins ; drug effects ; metabolism ; Rats ; Rats, Wistar ; Sciatic Nerve ; drug effects ; metabolism ; physiopathology
10.Replication of Bombyx mori Densonucleosis Virus (Zhenjiang isolate) in different silkworm strains.
Xu HAN ; Qin YAO ; Lu GAO ; Yong-Jie WANG ; Fang BAO ; Ke-Ping CHEN
Chinese Journal of Biotechnology 2007;23(1):145-151
The invertebrate parvovirus Bombyx mori Densonucleosis Virus type 3(Zhenjiang isolate), named BmDNV-3, is a kind of bidensovirus. The most obvious characteristic in the genome of BmDNV-3 is that it has 2 sets of DNA molecular (VD1, VD2),and each of them is encapsidated respectively in the form of single-stranded liner DNA ( + VD1, - VD1, + VD2, - VD2) in equal percentage. So the BmDNV-3 has 4 kinds of virions. Furthermore the sequence of BmDNV-3 is able to encode DNA polymerase itself. Some strains of silkworm revealed complete resistance to BmDNV-3, so they didn' t fall sick. To investigate the difference in the process of infection and replication between the 2 virions ( VD1, VD2) of this bidensovirus, and the difference of the increment in the resistant or susceptible host, the 5th instar larvae of the susceptible silkworm strain (HUABA 35) and the resistant silkworm strain(QIUFENG d) were inoculated determinate dose of BmDNV-3 by oral ingestion. Then the midgut were collected at 9 timepoints. The silkworm cytoplasm actin A3 was used to be normalized gene, so the number of cells in collected tissue could be determined. The result shows that whatever in the susceptible silkworm strain or in the resistant one, the copies of VD1 and VD2 in the genome of BmDNV-3 collected at the different timepoint were almost at the equal level respectively, so that the VD1 and VD2 were replicated with synchronization. The process of infection in the susceptible silkworm strain was devided into 3 partitions, latent period( 2 - 12 hours post inoculation), exponential phase (12 - 36 hours post inoculation)and stationary phase (36- 96 hours post inoculation and there are about 2 x 10(5) copies per cell) . In the resistant silkworm strain, the virus were replicated at a very low level, that was from 6 - 10 copies 2 hours post inoculation to 150 - 200 copies 96 hours post inoculation (about 20 times) . So we predict that the resistance in some of the silkworm strains from BmDNV-3 was a kind of chronic representation that the host carried virus without being caused flacherie.
Animals
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Bombyx
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genetics
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virology
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DNA, Viral
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genetics
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Densovirus
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genetics
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physiology
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Female
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Genome, Viral
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genetics
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Host-Pathogen Interactions
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Male
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Polymerase Chain Reaction
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methods
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Time Factors
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Virion
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genetics
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physiology
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Virus Replication
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genetics