Detecting testis- and epididymis-specific methylated promoters in human cell-free seminal DNA by MeDIP-qPCR / 中华男科学杂志

<p><b>OBJECTIVE</b>To establish a method of methyl-DNA immunoprecipitation (MeDIP)-real time quantitative PCR (qPCR) for detecting the promoter methylation level in cell-free seminal DNA (cfsDNA).</p><p><b>METHODS</b>We obtained cfsDNA samples from 6 normozoospermia men (the NZ group) and 6 post-vasectomy patients (the PV group), and mixed the samples from different individuals of each group, respectively. Then we made DNA fragments by ultrasonication, separated the methylated DNA fragments by MeDIP, and determined the methylation level of the promoters in cfsDNA by qPCR.</p><p><b>RESULTS</b>The methylation levels of the promoters PRAME, PEG10, MORC1, GML, HOXA5, DNMT3L, SNURF, MSH4, DAZ1 and CLPB were 14.93, 2.64, 0.69, 2.66, 17.50, 21.10, 5.98, 2.28, 13.50 and 3.86%, respectively, in the NZ group, obviously lower than 121.25, 73.62, 16.25, 42.90, 76.74, 112.40, 59.79, 25.85, 91.90 and 64.53% in the PV group. The results of MeDIP-qPCR for the methylation of PRAME, MORC1, GML, HOXA5, DNMT3L, SNURF, MSH4 and DAZ1 were coincident with the results of genome-wide promoter methylation microarray.</p><p><b>CONCLUSION</b>MeDIP-qPCR can quantitatively measure the promoter methylation level in cfsDNA, and effectively determine the testis- and epididymis-specific methylated promoters in human semen.</p>

Advances in the studies of cell-free seminal DNA / 中华男科学杂志

Cell-free DNA, also referred to as extracellular DNA, has been detected in many kinds of human body fluids, including blood plasma, urine, cerebrospinal fluid, bronchoalveolar lavage fluid, amniotic fluid, and seminal plasma. At present, cell-free DNA has been reported widely as promising noninvasive biomarkers for disease diagnosis and research. Recent years have witnessed some progress in the studies of the general characteristics of cell-free DNA, such as its concentration, extent of molecular weight, origin and existing forms, as well as in its clinical application. Cell-free seminal DNA has been proposed as promising noninvasive biomarkers for the studies and diagnosis of male idiopathic infertility, and the early diagnosis, treatment evaluation and outcome prediction of testicular germ cell tumors and prostatic cancer. This review summarizes the general characteristics and biological functions of cell-free DNA, and outlines the research status and application perspective of cell-free seminal DNA.

Expression patterns of age-related molecule c-kit, HIWI and vimentin in the rat testis and epididymis / 中华男科学杂志

<p><b>OBJECTIVE</b>To examine the differences in the expressions of c-kit, HIWI and vimentin in the testis and epididymis of rats aged different months, investigate the regular changes in the expression of characteristic molecules in the ripening and senescent process of the testis and epididymis, and provide experimental evidence for studies on the aging of the male reproductive system.</p><p><b>METHODS</b>Sprague-Dawley male rats were divided into a young group (6-month-old, n=10), an adult group (12-month-old, n=10) and an aged group (24-month-old, n=10). The immunohistochemical SP method was used to examine c-kit, HIWI and vimentin in the testis and epididymis of the rats.</p><p><b>RESULTS</b>The positive immunohistochemical reaction to c-kit was observed mainly in spermatogonia, weakly in other cells and also in the epithelia and lumens of the epididymis. The abundant expression of HIWI was detected in all stages of spermatogenic cells in the testis and epididymis of the 6-month-old and 12-month-old rats. Moreover, spermatogonias and primary spermatocytes displayed intense expression. Sertoli cells, Leydig cells, myoid cells and vascular endothelial cells (VEC) were also HIWI-positive cells. But the positive expression decreased with age in the testis and epididymis of the 24-month-old rats (P > 0.05). vimentin expression was weak in the testis and epididymis of the 6-month-olds but increased significantly in the 24-month-olds (P > 0.01).</p><p><b>CONCLUSION</b>The expressions of c-kit and HIWI decreased in the testis and epididymis of the 24-month-old rats, while vimentin expression increased markedly with age. The results suggest that the aging of the testis and epididymis is closely related to the abnormal transduction of cell signals.</p>

Analysis of the differences in the expression of HSP27 and c-kit between benign prostatic hyperplasia and prostatic cancer tissues / 中华男科学杂志

<p><b>OBJECTIVE</b>To examine the differences in the expression of HSP27 and c-kit between benign prostatic hyperplasia (BPH) and prostatic cancer (PCa) tissues and to analyse the relationship between their expression and BPH and PCa, especially the relationship with the occurrence, development, prognosis and treatment of PCa.</p><p><b>METHODS</b>An immunohistochemical staining (SP method) for HSP27 and c-kit was undertaken on 40 BPH and 40 PCa tissues samples.</p><p><b>RESULTS</b>Consistent patterns of cytoplasmic staining for HSP27 were seen in all sections of tissue from BPH. The glandular epithelium stained very strongly positively and the stroma stained positively. The staining for HSP27 in PCa tissues was located in the cytoplasm of glandular epithelia, but the expression of HSP27 in PCa was higher than BPH (P < 0.05). The staining for c-kit in BPH tissues was located in the cytoplasm of smooth muscle cells, and in PCa tissues was located in epithelial cells. The expression of c-kit in PCa tissues was lower than BPH (P < 0.05). The expression level of both HSP27 and c-kit were decreased with the development of grade of PCa (P < 0.05); HSP27 was increased with the development of clinical stage of PCa (P < 0.05 ); c-kit was decreased with the development of clinical stage of PCa (P < 0.05).</p><p><b>CONCLUSION</b>The expression level of HSP27 and c-kit was highly correlated with the process of the development from BPH to PCa, and also correlated with tumor grades and stages. The expression of HSP27 and c-kit may be used as an important pathological index and may be helpful for the treatment of PCa.</p>

Effect of eucommia ulmoides oliv on SOD activity and alpha-actin expression in the penile tissues of diabetic rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the effect of aqueous abstract from eucommia ulmoides oliv on the activity of superoxide dismutase (SOD) and alpha-actin expression in the penile tissues of rats with diabetes mellitus (DM) in vitro.</p><p><b>METHODS</b>A diabetes model was established by administration of alloxun twice to Sprague Dawley (SD) rats. Ten diabetic and 10 normal rats were randomly selected and the penile strips of each rat were divided into four equal shares and cultured in two groups, a eucommia ulmoides oliv coculture group (Group A, further dicided into 1 microg/ml, 10 microg/ml and 100 microg/ml subgroups) and a control group (Group B). Seven days later, the activity of SOD in the culture medium was detected by spectrophotometry, and the levels of micro-actin expression in the penile tissues were examined with the immunohistochemical method.</p><p><b>RESULTS</b>Compared with Group B, the activity of SOD in the culture medium in athe 10 and 100 microg/ml subgroups was notably elevated (P < 0.01), and the numbers of immunoreactive positive cells of alpha-actin in the penile tissues remarkably increased (P < 0.01).</p><p><b>CONCLUSION</b>The activity of SOD and alpha-actin expression in the penile tissues of diabetic rats in vitro can be increased by eucommia ulmioides oliv.</p>

Effect of L-methionine on trace elements in lead-intoxicated mice / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the effect of L-methionine (L-Met) on the content of Zn, Cu, Mn, Fe in liver, brain, spleen and kidney of lead intoxicated mice.</p><p><b>METHODS</b>Distilled water was given to 10 mice (normal control group) and lead acetate solution of 400 micro g/ml Pb(2+) to 20 mice to serve as drinking water for 10 days. The lead administration was then withdrawn and lead exposed mice were randomly divided into two groups: the lead control group took distilled water as drinking water for 4 weeks to serve as positive control, the other one took L-Met solution (0.5 mg/ml) as drinking water for 4 weeks (Pb + L-Met group) to serve as the treatment group. All the animals were sacrificed on the 1st day after 4 weeks, and the contents of Zn, Cu, Fe, Mn, Pb in liver, brain, spleen and kidney were measured by Inductively Coupled Plasma (ICP) Emission Spectrometry.</p><p><b>RESULTS</b>Lead contents in liver, brain, spleen and kidney of Pb control group [(1.490 +/- 1.654) micro g/g, (3.470 +/- 2.757) micro g/g, (4.975 +/- 2.993) micro g/g, (0.066 +/- 0.001) micro g/g respectively], were higher than those in normal control group [(0.015 +/- 0.001) micro g/g, (0.009 +/- 0.007) micro g/g, (0.027 +/- 0.002) micro g/g, (0.006 +/- 0.015) micro g/g, P < 0.05] while Zn contents in liver, brain, spleen and Fe and Mn content in liver, brain, spleen and kidney in Pb control group were lower than those in normal control group (P < 0.05). Pb contents of brain, spleen and Cu content of kidney in Pb + L-Met group were higher than those in normal control group (P < 0.05). Zn contents of liver, brain, spleen, Fe contents of liver, brain, spleen, kidney, and Mn contents of brain, spleen in Pb + L-Met group were lower than those in normal control group (P < 0.05). Fe contents of liver, brain, Zn content of spleen, Cu content of kidney and Mn contents of liver, brain, spleen in the Pb + L-Met group were higher than those in the Pb control group (P < 0.05). The lead levels of four organs in the Pb + L-Met group were lower than those in the Pb control group (P < 0.05).</p><p><b>CONCLUSION</b>Lead could be eliminated by L-Met, which may affect the distribution and metabolism of trace elements in mice.</p>

Urodynamic study of neurogenic bladder model after spinal cord injury in rat / 中华物理医学与康复杂志

Objective To observe the urodynamic change after spinal cord injury at different levels and the relationship with neurogenic dysfunction of bladder and urethra. Methods Eighty female rats were divided into a control group (20 rats) , a suprasacral spinal cord injury group (30 rats) and a sacral spinal cord injury (30 rats). The urodynamic exam was performed with all the rats before and 20 days after the spinal cord injury model was established by surgical operation. Results The maximum bladder volume and compliance in the su- prasacral injury group were significantly less than the sacral spinal cord injury group and the control, the maxi- mum volume and compliance in sacral spinal cord injury group were significantly less than the control. The DLPP in suprasacral injury group was significantly higher than that in the sacral spinal cord injury group and the con- trol, the DLPP in sacral spinal cord injury group was significantly less than that in the control group. Conclu- sion Urodynamic study is very useful for the early diagnosis and individualized treatment of the neurogenic bladder after spinal cord injury.