Effects of Rho-associated coiled-coil-forming kinase inhibitor Y27632 on mouse preantral follicles development in vitro / 解剖学报

Objective To investigate the effects of Rho-associated coiled-coil-forming kinase (ROCK) inhibitor Y27632 on the development of mouse preantral follicles in vitro. Methods The ovaries of 12.5-day-old mice were collected and single preantral follicle was isolated by mechanical method to culture Nunclon Sphera 96-well U-bottom 3D cell culture plate in vitro. Set up a control group and treated group containing Y27632, and evaluate the overall development of the follicles through follicular morphology, follicle diameter, the number of mature follicles, and changes in oocyte spindles. The expression of oocytes [bone morphogenetic protein 15(BMP 15), growth differentiation factor 9(GDF9)], granulosa cells [follicle stimulating hormone receptor (FSHR)] and apoptosis related genes (Bad, Bax and Caspase-3) were also assessed by the Real-time PCR technique. Meanwhile, the follicle viability was detected by follicular activity test during follicular development. Results Rock inhibitor Y27632 had no significant effect on the growth diameter and basic development indicators (follicle survival rate, cavity formation rate and maturation rate) (P>0.05). And abnormal spindle assembly occurred during follicle development in the control group. After 8 days of culture, compared with the control group, the granulosa cell-specific gene FSHR was up-regulated in the experimental group; As well as the oocyte-specific genes BMP 15 and GDF9 were up-regulated, while forkhead box 03(Fox03) was down-regulated; The apoptosis genes Bax, Bad and Caspase-3 were also showed down-regulated. And the granulosa cell apoptosis in the experimental group was significantly less than that in the control group. Conclusion ROCK inhibitor Y27632 can inhibit the apoptosis of granulosa cells and prevents the abnormal assembly of oocyte spindles to improve the quality of follicle development in vitro.

Effects of single heat stress treatment on spermatogenic cells in mice / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of single heat stress treatment on spermatogenic cells in mice.</p><p><b>METHODS</b>We randomly divided 36 C57 male mice into a control and a heat stress treatment group and submerged the lower part of the torso in water at 25 °C and 43 °C, respectively, both for 15 minutes. At 1, 7, and 14 days after treatment, we obtained the testicular organ indexes, observed the changes in testicular morphology by HE staining, and determined the location and expression levels of the promyelocytic leukemia zinc finger (PLZF) and synaptonemal comlex protein-3 (SCP-3) in the testis tissue by immunohistochemistry and Western blot.</p><p><b>RESULTS</b>The testicular organ index was significantly lower in the heat stress treatment than in the control group (P < 0.05). Compared with the controls, the heat shock-treated mice showed loosely arranged spermatogenic cells scattered in the seminiferous tubules at 1 day after heat stress treatment, atrophied, loosely arranged and obviously reduced number of spermatogenic cells at 7 days, and relatively closely arranged seminiferous tubules and increased number and layers of spermatogenic cells at 14 days. The number of SCP-3 labelled spermatocytes obviously decreased in the heat stress-treated animals at 1 and 7 days and began to increase at 14 days. The PLZF protein expression was significantly reduced in the heat stress treatment group at 1 day as compared with that in the control (0.19 ± 0.12 vs 0.64 ± 0.03, P < 0.01), but elevated to 0.77 ± 0.02 at 7 and 14 days, even remarkably higher than in the control animals (P < 0.01).</p><p><b>CONCLUSION</b>Heat stress treatment can induce short-term dyszoospermia in mice, which can be recovered with the prolonged time after treatment.</p>

Dynamic study and screening of new markers of spermatogonial stem cells by iTRAQ protein mass spectrometry / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the dynamic changes in the protein marker expression in the spermatogonial stem cells (SSCs) of mice at different ages by iTRAQ protein mass spectrometry and to screen new markers using the bioinformatic proteome database.</p><p><b>METHODS</b>Based on the postnatal weeks, we divided 80 healthy male C57BL/6 mice into eight age groups of equal number, harvested their testicular tissues, extracted proteins following purification of the SSCs by compound enzyme digestion and magnetic-activated cell sorting. Then we analyzed and identified proteins using two-dimensional electrophoresis, protein mass spectrometry, and protein database information.</p><p><b>RESULTS</b>Totally, 248,510 mass spectra were obtained from the MS experiment and 1132 proteins were identified. By the criteria of >1.2-fold for protein abundance difference and P value <0.05, we identified 298 differentially expressed proteins and 9 currently known makers of SSCs (PCNA, GFRalpha1, CDH1, Annexin A7, UCHL1, VASA, CD49f, CD29, and PLZf). Compara- tive analysis showed different expressions of the proteins in the SSCs of the mice of different ages, and the differences in the expressions of GFRalpha1, CD49f, and CD29 were consistent with the findings in other published literature. Ten proteins (P63, CD71, CD98, K19, ACE, K18, K15, K17, SH2, and SH3) were selected as SSC markers to be further studied.</p><p><b>CONCLUSION</b>The proteins in SSCs are differentially expressed in mice of different ages. The technology of iTRAQ protein mass spectrometry can be used to analyze and compare the proteome information of mouse SSCs, obtain differentially expressed proteins in mice of different ages, and thus offers a new ap- proach to further analysis and study of the function and roles of these differential proteins.</p>

A case-control study on risk factors for tuberculosis in Yinchuan city / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the risk factors of tuberculosis in Yinchuan city and lay a basis for its prevention and control.</p><p><b>METHODS</b>A matched case-control (119:179) study for the risk factors was carried out. Data were analyzed with single-variable analysis and multiple factor logistic regression analysis.</p><p><b>RESULTS</b>Single-variable analysis showed that the education background (chi2 = 2.363, P = 0.018), family economic income (chi2 = 3.040, P = 0.002), smoking (chi2 = 2.500, P = 0.012), physical activities (chi2 = 2.330, P = 0.020), bacille Calmette-Guerin (BCG) vaccination history (chi2 = 22.151, P = 0.000), history of exposure to tuberculosis (chi2 = 15.740, P = 0.000) and so on had significant effects on tuberculosis. Multiple logistic regression analysis showed that family monthly income, smoking, physical activity, BCG vaccination history, history of exposure to tuberculosis entered the final regression model (chi2 = 5.880, 7.368, 3.891, 21.127, 14.536; OR = 0.529, 1.571, 0.774, 0.264, 3.978; P < 0.05).</p><p><b>CONCLUSION</b>History of exposure to tuberculosis and smoking should be the risk factors of tuberculosis in Yinchuan. Having much income, physical activities, and BCG vaccination history should be the preventive factors.</p>

DNA fingerprinting of Mycobacterium tuberculosis strains from Beijing, Guangdong and Ningxia / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the epidemic distribution of Mycobacterium tuberculosis isolates from Beijing, Guangdong and Ningxia, and to determine M. tuberculosis strains of the "Beijing Family".</p><p><b>METHODS</b>Two hundred and six IS6110 DNA fingerprinting patterns of M. tuberculosis strains from three provinces (city) were transferred to digital data, compared with the world M. tuberculosis DNA fingerprinting database, and then clustered by Gel compare 4.1 software. The clustering values in different patients with tuberculosis were compared by chi(2) test. Risk factors for recent transmission were calculated using odd ratios.</p><p><b>RESULTS</b>No M. tuberculosis strains were found the same as those of DNA fingerprint database. 56.8% (117/206) fingerprinting patterns of M. tuberculosis shared by least two-thirds of the IS6110 fragments and their Spoligotyping fingerprinting patterns were consistent with those of M.tuberculosis strains of the "Beijing Family". There were significant differences between female and male, different age groups (< 42 years old) and older (>or= 42 years old) (P < 0.05). Odd ratio was 5.06 in the group younger than 42 years old (95% CI: 1.00 - 34.34) and was 4.43 (95% CI: 0.94 - 28.76) in males.</p><p><b>CONCLUSION</b>M. tuberculosis strains of "Beijing Family" were popular in Beijing, Guangdong and Ningxia. Men and younger age group (< 42) were shown to be infected by identical strains more often than women and older aged which might play an important role in the recent transmission of tuberculosis in these areas. IS6110 DNA fingerprinting of M. tuberculosis could be used to trace the source of tuberculosis infection.</p>