Standardization and quality control for determination of fructose in seminal plasma.
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Centrifugation↗
Biomedical subjects
Jin-Chun Lu
Publications and source records attributed to Jin-Chun Lu.
Preliminary investigations on the standardization and quality control for the determination of acid phosphatase activity in seminal plasma.
BACKGROUND: The determination of ACP in seminal plasma was considered as an appropriate biochemical marker to evaluate prostate function, as recommended by the WHO manual. However, few reports on the standardization and quality control for the determination of biochemical markers in seminal plasma have been documented. METHODS: Two frozen samples of seminal plasma with or without phenylmethylsulfonyl fluoride were determined for their acid phosphatase (ACP) levels. The ACP level and sperm concentration of each of 72 samples of seminal plasma obtained at 1000xg for 10 min or 3000xg for 15 min centrifugation were assayed. ACP activity in 10 samples of seminal plasma was measured immediately or standing for 30 min after dilution. The ACP levels in seminal plasma with or without chymotrypsin were also assayed. RESULTS: There was no significant difference of ACP levels (P=0.166) but of sperm concentrations (P=0.000) in seminal plasma obtained by centrifugation at different velocity. ACP activities in seminal plasma measured when standing for 30 min after dilution were significantly lower than those measured immediately after dilution (P=0.001). Both chymotrypsin and freezing-thawing had no apparent effect on the determination of ACP in seminal plasma. CONCLUSION: The results indicated that standing time after dilution and centrifugation velocity should be standardized, and frozen seminal plasma could serve as the quality control products for the determination of ACP activity among different laboratories.
Acid Phosphatase↗
Chymotrypsin effects on the determination of sperm parameters and seminal biochemistry markers.
BACKGROUND: Few reports of the effects of treatment with chymotrypsin on the determination of sperm parameters and seminal biochemistry markers are documented. METHODS: Sperm parameters of 63 liquefied and 27 non-liquefied samples, untreated or treated with chymotrypsin, were evaluated using computer-assisted semen analysis. In addition, biochemistry markers such as gamma-glutamyltranspeptidase, alpha-glucosidase and fructose in 50 liquefied and 39 non-liquefied samples, untreated or treated with chymotrypsin, were determined. RESULTS: Treatment with chymotrypsin had no effect on sperm concentration, motility, motility a and b, straightness, curvilinear velocity, straight line velocity, average path velocity and beat cross frequency in both liquefied and non-liquefied semen. However, linearity (p=0.025) decreased and the amplitude of the lateral head (p=0.029) increased significantly in non-liquefied semen after treatment with chymotrypsin. The levels of gamma-glutamyltranspeptidase, alpha-glucosidase and fructose in seminal plasma were unaffected by chymotrypsin, regardless of liquefaction status. CONCLUSIONS: Chymotrypsin had no effects on the detection of sperm parameters and biochemistry markers, and could be used to treat non-liquefied samples before semen analysis in the andrology laboratory.
Biomarkers↗
[Comparison and evaluation of two methods for measuring acid phosphatase activity in seminal plasma].
OBJECTIVE: To compare the routine method and the kit method for the measurement of acid phosphatase activity in seminal plasma, and to explore the possibility of the kit method for routine measurement. METHODS: Seventy-nine seminal plasma samples were assayed by routine method and kit method respectively for acid phosphatase. One sample was detected 10 times for within-run analysis, and an other two were measured by both the methods once a day for 10 days for between-run analysis. Acid phosphatase activities in another 10 seminal plasma samples collected at random were measured immediately or 30 min after dilution by two technicians, respectively. RESULTS: There were significant positive correlations between the acid phosphatase activities measured by routine and kit methods (r = 0.745, P = 0.000). In the within-run assay, the coefficient of variation for the kit method (13.72%) was similar with that for the routine method (10.66%). But in the between-run assay, the coefficients of variation for the kit method (13.8% and 15.49%) were obviously lower than those for the routine method (24.43% and 21.04%). Compared with the acid phosphatase activities in seminal plasma measured immediately after dilution, those measured after 30-min standing were notably lower for either of the methods (P < 0.05). However, there wasnt significant difference in the acid phosphatase activities detected by the routine method between the two technicians (P = 0.165). CONCLUSION: The kit method is superior and preferable to the routine method for the measurement of acid phosphatase in seminal plasma.
Acid Phosphatase↗
[Evaluation of the determination of seminal ACP and gamma-GT activities and correlation between seminal ACP or gamma-GT activity and semen parameters].
OBJECTIVE: To evaluate the determination of seminal acid phosphatase (ACP) and gamma-glutamyltranspeptidase (gamma-GT) activity, and analyze the correlation between seminal ACP or gamma-GT and semen parameters. METHODS: ACP and gamma-GT activities in 133 samples of seminal plasma were measured. Two of the samples were randomly selected for intra-assay, one for the detection of ACP activity and the other for gamma-GT activity. And another four were selected the same way for the same purpose, two for the detection of ACP activity and the other two for gamma-GT activity. The semen volume, pH, sperm concentration, motility, and grade-a and -b motility were analyzed by CASA system and so were the correlation between seminal ACP or gamma-GT activity and semen parameters. RESULTS: There was significant positive correlation between ACP and gamma-GT activities (r = 0.570, P = 0.000). The intra-CV of ACP was 13.72%, and inter-CVs of ACP were 13.80% and 15.49%. The intra-CV of gamma-GT was 7.68%, and inter-CVs of gamma-GT were 7.76% and 9.73%. Both seminal ACP and gamma-GT activities had significant negative correlation with pH (r = -0.330, P = 0.000 vs r = - 0. 388, P = 0.000). There was obvious correlation between gamma-GT activity and sperm concentration (r = 0.165, P = 0.045), but not between ACP activity and sperm concentration (r = 0.048, P = 0.546). Neither of seminal ACP and gamma-GT activity was correlated with sperm motility, grade-a and -b motility, semen volume, abstinence time and age. CONCLUSION: The precision of the measurement of gamma-GT activity in seminal plasma was higher than that of ACP. The correlation between seminal gamma-GT activity and semen parameters was similar to that between seminal ACP activity and semen parameters. Thus, the determination of gamma-GT activity was a more reliable marker than that of ACP activity for the evaluation of prostate function.
Acid Phosphatase↗
[Advances in the studies of Chlamydia trachomatis infection in males].
Chlamydia trachomatis(Ct) infection in reproductive tract is one of the most common sexually transmitted diseases at present. However, relatively fewer studies are made on Ct infection in men. The paper reviews the epidemiology of Ct infection in males, Ct infection and male diseases, Ct infection and male infertility, and the detection of Ct infection in men. It aims at providing a theoretical basis and a practical guide for the prevention and control of Ct infection in men.
Chlamydia Infections↗
[Cloning and sequence analysis of Chlamydia trachomatis heat shock protein 10].
OBJECTIVE: To obtain Chlamydia trachomatis heat shock protein (cHSP) 10 gene from clinical secretion samples. METHODS: cHSP10 gene was amplified from 20 cases of clinical secretion samples with positive gold-labelling by specific primers of cHSP10 and identified by sequence analysis. RESULTS: cHSP10 full-length gene was amplified from 1 of 20 cases of clinical secretion samples with positive gold-labelling. cHSP10 gene encoding 102 amino acids contains 306 bp, which nuclotide at position 194 changes from T to A, leading to the change of corresponding amino acid. CONCLUSIONS: cHSP10 gene may be cloned from clinical secretion samples with positive gold-labelling, which make it possible to further construct expression plasmid of recombinant cHSP10.
Antibodies, Bacterial↗
[Detection of human sperm morphology and acrosome reaction with Coomassie brilliant blue staining].
OBJECTIVES: To evaluate the application of Coomassie brilliant blue (CBB) G250 staining for the detection of human sperm deformity rate, rate of intact acrosome and acrosome reaction. METHODS: The smear of spermatozoa before and after capacitation and induced acrosome reaction with progesterone (P) were stained with 0.05% CBB G250 and Wright-Giemisa solution respectively, and visualized with light microscopy. The deformity rate of spermatozoa, rate of intact acrosome and acrosome reaction were calculated. RESULTS: There was no any difference in detection of deformity rate of spermatozoa and rate of intact acrosome with CBB G250 and Wright-Giemisa staining(P < 0.05). The sperm population of acrosome reaction with induced P was divided by CBB staining into two types: positive staining with dark violet blue on acrosome cap and pale or negative staining on the same area. The rate of the latter was increasing with increasing inductive time, maybe representative of the rate of acrosome reaction. The mean rate was(75.1 +/- 3.8)% after induced for 1 h. CONCLUSIONS: CBB G250 staining is a reliable method for assessment of the human sperm morphology and acrosome reaction.
Acrosome↗
[Preliminary study on hens immunized with recombinant human testis prostaglandin D synthase DNA].
OBJECTIVES: In order to identify whether the hens immunized with recombinant human testis prostaglandin D synthase (rhtL-PGDS) DNA can produce anti-L-PGDS antibody. METHODS: The serum were got from the hens immunized with recombinant plasmid pGEX-2T/htL-PGDS DNA (100 micrograms) every 2 weeks for 2 times. The exist of anti-L-PGDS antibody and its titer were tested with agarose dual immunodiffusion and ELISA with rhtL-PGDS as antigen. RESULTS: The serum anti-L-PGDS antibody in hen immunized with pGEX-2T/htL-PGDS DNA were confirmed and its titer tested by ELISA was 1:2,048. CONCLUSIONS: It is feasible to produce anti-L-PGDS antibody by immunizing hens with recombinant pGEX-2T/htL-PGES DNA.
Animals↗
[Advances in sperm capacitation].
Sperm must be capacitated before sperm-ovum fusion. Capacitation was once considered as hyperactivation. But now many investigators thought that capacitation wasn't equal to hyperactivation, and that sperm hyperactivation might be a moiety of capacitation or the result of capacitation. In the present, the methods used to study sperm capacitation include fertilization in vitro, induction of sperm acrosome reaction, FITC-labeled chlortetracycline and plant hemoagglutinin. The studies on sperm capacitation in vitro mainly focused on the inductive substances of sperm capacitation and subsequent results analysis. It could lay foundation for the manifestation of molecular mechanism of sperm capacitation and destination of sperm capacitation in molecular levels.
Adenylyl Cyclases↗