Significance of sRANKL/OPG ratio in diagnosis of multiple myeloma bone disease / 中国实验血液学杂志

This study was purposed to investigate the relationship between the levels of soluble receptor activator of nuclear factor kappa B ligand (sRANKL) and osteoprotegerin (OPG) in serum of the patients with multiple myeloma (MM) and multiple myeloma bone disease (MBD). The serum levels of sRANKL, OPG, tartrate-resistant acid phosphatase-5b (TRAP-5b) and C-terminal telopeptide of collagen I (CTP-I) which both are indexes for metabolism of osteoclast (OC) in newly diagnosed MM patients (n=42, experimental group) and healthy persons (n=25, control group) were detected by enzyme-linked immunosorbent assay. The roentgenography was used to determine bone damage in MM patients at the same time. According to these results acquired, the correlation of sRANKL/OPG ratio with levels of TRAP-5b/CTP-I, the incidence and degree of bone destruction were analyzed. The results indicated that the level of sRANKL (median value 9.33 microg/L) increased and level of OPG (median value 4.93 microg/L) decreased and the sRANKL/OPG ratio (2.65) increased significantly in experimental group. Compared with control group, the differences in all the corresponding indicators were statistically significant (p<0.05). The sRANKL/OPG ratio was closely related to levels of TRAP-5b (r=0.512, p<0.05) and CTP-I (r=0.481, p<0.05) in MM patients. After all patients in experimental groups were divided into group with bone destruction (n=29) and without bone destruction (n=13), the sRANKL/OPG ratio in the group with bone destruction was 5.13 and much higher than that in group without bone destruction (1.12) (p<0.05). A close correlation between the sRANKL/OPG ratio and degree of bone destruction (r=0.445, p<0.05) was acquired when all MM patients were divided into three groups according to degree of bone destruction, but no difference between the ratio and clinical classification and International Staging System (ISS) in MM patients was found. It is concluded that the sRANKL/OPG ratio in serum of MM patients is significantly elevated, which may be closely related to increase metabolism of OC along with the incidence and degree of bone destruction. In short, the sRANKL/OPG ratio can be used as a reference index for the diagnosis of MBD.

Primitive research on the effect of FSH autoantibody on the spermatogenic capability of the rat testis / 中华男科学杂志

<p><b>OBJECTIVE</b>To establish rat models of FSH autoantibody and to investigate the effect of FSH autoantibody on the spermatogenic capability of rat testis.</p><p><b>METHODS</b>Thirsty 21-day old SD rats were randomly divided into an experimental and a control group of equal number. A specific polypeptide corresponding to the rat FSHbeta subunit was synthesized and coupled to (keyhole limpet hemocyanin) KLH. The rats in the experimental group were immunized with polypeptide-KLH and these in the control group with KLH. Further immunization was performed every 2 weeks for 7 times. On the 77th, 91st and 105th day of the immunization, 5 rats from the experimental group and another 5 from the control group were killed. Then the structures of the seminiferous tubule and epididymal sperm were observed by light and electron microscope, respectively. Meanwhile, the counts of sperms and the percentage of swelled sperm were calculated. And the level of serum testosterone was detected by enzyme-linked immunospecific assay (ELISA).</p><p><b>RESULTS</b>The titer of the anti-polypeptide antibody was 1:200 on the 49th day of the immunization, and reached 1:400 on the 63rd. Compared with the control group, the percentage of swelled sperm significantly decreased on the 91st day (60.4 +/- 6.23 vs 50.60 +/- 3.05, P < 0.05), and the number of spermatogenic cells and sperms in seminiferous tubules reduced on the 105th day in the experimental group, the counts of sperms (46.08 +/- 6.56 vs 32.53 +/- 3.41) and the percentage of swelled sperm (60.60 +/- 5.86 vs 48.60 +/- 3.85) significantly lower (P < 0.05), while the level of serum T significantly higher than that in the control group (P < 0.05).</p><p><b>CONCLUSION</b>FSH autoantibody might cause testis dyszoospermia.</p>

Determination of alpha-glucosidase activity in seminal plasma by semi-automatic biochemistry analyzer / 中华男科学杂志

<p><b>OBJECTIVE</b>To establish a method of determining alpha-glucosidase activity in seminal plasma by semi-automatic biochemistry analyzer.</p><p><b>METHODS</b>Alpha-glucosidase activity in seminal plasma from 51 men with normal semen parameters in routine semen analysis were detected by semi-automatic biochemistry analyzer and manual glucose oxidase method, respectively. Then, the intra-assay and inter-assay coefficient variation (CV) and normal reference value were calculated. In the meanwhile, the correlation between the two methods was analyzed.</p><p><b>RESULTS</b>The intra-assay CVs of 2 seminal plasma samples with different alpha-glucosidase activity detected by semi-automatic biochemistry analyzer were 12.63% and 9.13%, and the inter-assay CVs were 10.67% and 13.49%, respectively. The normal reference value for seminal alpha-glucosidase activity detected with semi-automatic biochemistry analyzer ranged from 102.28 to 555.08 U/L. There was a significantly positive correlation between the semi-automatic biochemistry analyzer and the manual glucose oxidase method (r = 0.792, P < 0.01).</p><p><b>CONCLUSION</b>The method of determining alpha-glucosidase activity in seminal plasma by semi-automatic biochemistry analyzer, with its simplicity, less cost of time and reagents, and more reliable result, could be applied to clinical laboratory medicine.</p>

Primary investigations on the quality control for semen analysis in Nanjing City / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate and analyze the results of the determination of sperm concentration, fructose concentration, alpha-glucosidase and acid phosphatase (ACP) activities in the seminal plasma from different hospitals in the city of Nanjing, so as to provide a basis for the external quality control (EQC) of semen analysis within Jiangsu Province or even the whole country.</p><p><b>METHODS</b>Eight samples of quality control products for low and high concentrations sperm count, fructose, alpha-glucosidase and ACP determination were prepared and divided, each detected for the sperm concentration, fructose, alpha-glucosidase and ACP activity, and the coefficient variances (CVs) were calculated. The products were then distributed to 11 hospitals in the city, and the results were collected and analyzed. In addition, the total relative errors (REs) for each product was calculated based on the results after dividing as reference values.</p><p><b>RESULTS</b>The CVs from the 8 samples after dividing were 3.83% - 11.16%. Collected from the 11 hospitals attending EQC were 11 reports of the results of sperm concentration, and 5 the results of fructose, alpha-glucosidase and ACP in seminal plasma. Among the results from different laboratories, those of fructose determination showed the minimal difference (CVs: 8.99% and 3.95% for low and high concentrations, respectively) , next came alpha-glucosidase (CVs: 16.66% and 18.41% for low and high activities, respectively), and ACP determination showed the maximal difference (CVs: 54.12% and 65.58% for low and high activities, respectively). Moreover, the same trend was observed in RE values, as shown in the total REs, which were 11.99% (low concentration) and 20.31% (high concentration) for the determination of fructose in seminal plasma, 22.92% and 27.26% for alpha-glucosidase, 7.34% and 318.35% for ACP in different laboratories, and the maximal RE value was detected in the result of the high-activity ACP sample. Of the 11 hospitals, 6 determined sperm concentration with the computer-assisted semen analysis (CASA) system, and the other 5 with the modified hemocytometer. RE values (148.47% and 187.59% for low and high concentration samples, respectively) and sperm concentrations ([62.74 +/- 16.63] x 10(6)/ml and [163.32 +/- 36.24] x 10(6)/ml) counted with the hemocytometer were significantly higher than those with the CASA system (REs 13.97% and 10.48%; sperm concentrations [24.88 +/- 4.16] x 10(6)/ml and [54.24 +/-23.06] x 10(6)/ml ).</p><p><b>CONCLUSION</b>The methods of seminal alpha-glucosidase and fructose determination were relatively stable in current andrology laboratories, and the variance range could be accepted. However, the method of seminal ACP determination might be unadaptable to clinical application, and needs to be further improved. Hemocytometer, which significantly overestimated sperm concentration, could not be applied to the assay of sperm concentration.</p>

Application of distilled water in sperm counting and hypoosmotic swelling test / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the application of distilled water in sperm-counting and hypoosmotic swelling test.</p><p><b>METHODS</b>Thirty-seven semen samples were collected and each was diluted by distilled water and sodium acid carbonate-formaldehyde solution, respectively. Then the hemacytometer was used for sperm counting. Meanwhile, the percentage of swelled sperm diluted by distilled water was compared with the result of hypoosmotic swelling test recommended by WHO. Another 26 semen samples were diluted by distilled water and hypoosmotic swelling solution respectively, and the percentages of the swelled sperm were compared.</p><p><b>RESULTS</b>There was no significant difference either between the sperm concentrations obtained by distilled water and sodium acid carbonate-formaldehyde solution (P > 0.05) or between the percentages of the swelled sperm diluted by distilled water and hypoosmotic swelling solution.</p><p><b>CONCLUSION</b>Distilled water can not only replace sodium acid carbonate-formaldehyde solution for sperm-counting dilution but also be used as a hypoosmotic swelling solution.</p>

Comparison and evaluation of two methods for measuring acid phosphatase activity in seminal plasma / 中华男科学杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>The kit method is superior and preferable to the routine method for the measurement of acid phosphatase in seminal plasma.</p>

Evaluation of the determination of seminal ACP and gamma-GT activities and correlation between seminal ACP or gamma-GT activity and semen parameters / 中华男科学杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>