Novel methods for studies of testicular development and spermatogenesis: From 2D to 3D culture / 中华男科学杂志

The two-dimensional model of cell culture is an important method in the study of testicular development and spermatogenesis but can not effectively mimic and regulate the testicular microenvironment and the whole process of spermatogenesis due to the lack of relevant cell factors and the disruption of a three-dimensional spatial structure. In the past 20 years, the development and optimization of the in vitro model such as testis organotypic culture and in vivo model such as testis transplantation achieved a transformation from two- to three-dimension. The maintenance and optimization of the testicular niche structure could mimic the testicular microenvironment and cell types including Leydig, Sertoli and germ cells, which showed similar biological behaviors to those in vivo. Besides, the cell suspension or tissue fragment floats in the gas-liquid interface so that the development of somatic and germ cells is well maintained in vitro whilst the feedback linkage between grafted testis tissue and hypothalamus-pituitary of the host rebuilt in the in vitro model provides an endocrinological basis for spermatogenesis, which serves as an effective methodology to better understand the organogenesis and development of the testis as well as testicular function regulation, advancing the concept of treatment of male infertility. Al- though each of the methods may have its limitations, the progress in the processing, freezing, thawing, and transplantation of cells and tissues will surely promote their clinical application and present their value in translational medicine.

Seminal vesiculoscopy in the treatment of refractory hemospermia and ejaculatory duct obstruction / 中华男科学杂志

<p><b>Objective</b>To summarize the experience in the diagnosis and treatment of refractory hematospermia and ejaculatory duct obstruction by seminal vesiculoscopy.</p><p><b>METHODS</b>We retrospectively analyzed the clinical data about 42 cases of refractory hematospermia and 6 cases of ejaculatory duct obstruction with azoospermia. We investigated the diagnosis, treatment, and prognosis of the diseases.</p><p><b>RESULTS</b>All the patients underwent pelvic MRI and seminal vesiculoscopy. MRI for the 42 refractory hematospermia patients showed that 21 (50.0%) had cystic dilatation in the uni- or bilateral seminal vesicles, 25 (59.5%) had abnormal internal signal intensity in the uni- or bilateral seminal vesicles, 12 (28.6%) had both the problems above, and 4 (9.52%) had no obvious abnormality in the seminal vesicle area. The bilateral seminal vesicles were <1 cm in width in 3 of the 6 cases of ejaculatory duct obstruction, and obviously enlarged in the other 3, but without abnormal internal signals. No recurrence was found during the 3－36 months follow-up.</p><p><b>CONCLUSIONS</b>The history and physical examination play important roles in the diagnosis of refractory hemospermia, and MRI is more valuable than TRUS in the diagnosis of seminal vesicle diseases. Seminal vesiculoscopy is an effective option for the management of persistent hematospermia and ejaculatory duct obstruction.</p>

Spermatic vein ligation and intra-vaginal ejaculation latency / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the intra-vaginal ejaculation latency time (IELT) of varicocele patients, the influence of spermatic vein ligation on IELT, and the relationship of Visual Analogue Score (VAS) with IELT.</p><p><b>METHODS</b>We selected 112 males who had regular sexual life after spermatic vein ligation and conducted follow-up visits for 6 months. According to preoperative IELT, we divided the patients into an IELT < or = 2 min group and an IELT > 2 min group, and compared their IELT, VAS and Chinese Index of Sexual Function for Premature Ejaculation-5 (CIPE-5) scores before and 6 months after operation.</p><p><b>RESULTS</b>Follow-up was accomplished in 81 of the patients, 18 in the IELT < or = 2 min group and 63 in the IELT >2 min group. Compared with the baseline, IELT was significantly prolonged postoperatively in both the IELT < or = 2 min group ([1.26 +/- 0.37] vs [4.53 +/- 1.69] min, P < 0.01) and the IELT >2 min group ([5.14 +/- 2.03] vs [7.69 +/- 4.51] min, P < 0.05); the postoperative CIPE-5 scores were remarkably improved in the former (11.27 +/- 3.52 vs 15.64 +/- 2.37, P < 0.05) but insignificantly in the latter group (20.42 +/- 4.65 vs 21.83 +/- 5.49, P > 0.05); the postoperative grades of the CIPE-5 scores showed significant differences in both groups (chi2 = 6.353, P = 0.042 and chi2 = 3.910, P = 0.048); the postoperative VAS was markedly increased (3.18 +/- 0.92 vs 1.56 +/- 0.83 and 3.24 +/- 0.95 vs 1.74 +/- 0.79, P < 0.05), with significant differences in the grades of VAS in both groups (chi2 = 4.433, P = 0.035 and chi2 = 10.088, P = 0.001). The variation of VAS was negatively correlated with that of IELT in both groups (r = -0.572, P < 0.01 and r = -0.465, P < 0.05).</p><p><b>CONCLUSION</b>Varicocele may be one of the causes of premature ejaculation, and some of the varicocele patients with IELT < or = 2 min may benefit from spermatic vein ligation. Improved VAS is negatively correlated with prolonged IELT. The relationship between varicocele and premature ejaculation deserves further studies.</p>

Sexual development characteristics and sex hormone levels in obese male adolescents / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the characteristics of sexual development and sex hormone levels in obese male adolescents.</p><p><b>METHODS</b>We included 156 obese male adolescents with micropenis and microorchidia in an observation group and 50 healthy ones in a control group. We measured the body mass index (BMI), penile natural length and testicular volume, investigated the incidence of spermatorrhea and the age of the first spermatorrhea, detected the levels of serum luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin (PRL), total testosterone (TT), free testosterone (FT), progesterone (P) and estradiol (E2) using radioimmunoassay, and calculated TT/E2 and testosterone secretion index (TSI).</p><p><b>RESULTS</b>Compared with the healthy controls, the obese adolescents showed significantly higher BMI ([20.4 +/- 1.6] vs [27.1 +/- 2.2] kg/m2, P < 0.05), but shorter penile natural length ([6.7 +/- 2.1] vs [5.6 +/- 1.7] cm, P < 0.05) and lower testis volume ([9.9 +/- 3.1] vs [7.6 +/- 2.3] cm3, P < 0.05). The incidence of spermatorrhea was significantly decreased in the observation group in comparison with that of the control (chi2 = 17.335, P < 0.05), but there was no significant difference in the age of the first spermatorrhea between the two groups (P > 0.05). The levels of LH, E2 and P were remarkably higher in the observation group than in the control ([7.82 +/- 2.14] vs [5.39 +/- 1.76] mIU/ml, P < 0.05; [48.57 +/- 8.34] vs [8.61 +/- 4.08] pg/ml, P < 0.01; and [1.25 +/- 0.58] vs [0.64 +/- 0.19] ng/ml, P < 0.05), while TT and FT were markedly lower in the former than in the latter ([0.73 +/- 0.20] vs [1.47 +/- 0.41] ng/ml, P < 0.01 and [5.09 +/- 2.60] vs [11.28 +/- 4.72] pg/ml, P < 0.01), and so were the TT/E2 ratio and TSI (0.015 +/- 0.004 vs 0.173 +/- 0.037 and 0.098 +/- 0.026 vs 0.272 +/- 0.084, P < 0.01). BMI was correlated positively to PRL and E2, but negatively to TT, FT, TT/E2 and TSI (P < 0.05); the penile natural length positively to TT, FT, TT/E2 and TSI, but negatively to E2 (P < 0.05); and the mean testis volume positively to TT, FT, TT/E2 and TSI, but negatively to LH, PRL and E2 (P < 0.05).</p><p><b>CONCLUSION</b>Testis dysplasia and alteration of sex hormone levels exist in obese male adolescents. Obesity and fat accumulation lead to increased E2 and decreased TT and FT, particularly the reduction of TT/E2 and TSI, which suggest that the body fat content has an important influence on the development of the male reproductive system.</p>

Qianlieping capsule plus alpha-blocker for chronic non-bacterial prostatitis: analysis of 220 cases / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the clinical effects of Qianlieping Capsule combined with alpha-receptor blocker tamsulosin on chronic non-bacterial prostatitis (CNBP).</p><p><b>METHODS</b>We assigned 220 CNBP patients to three groups to receive oral Qianlieping Capsule (2.0 g tid) plus alpha-receptor blocker tamsulosin (0.2 mg qd) (n = 98), Qianlieping Capsule alone at 2.0 g tid (n = 66), and tamsulosin alone at 0.2 mg qd (n = 56) , respectively. After 6 weeks of medication, we assessed the therapeutic effects according to the NIH-CPSI scores and the number of small particles of lecithin (SPL) in the prostatic fluid after treatment.</p><p><b>RESULTS</b>Qianlieping Capsule alone increased the number of SPL by 46.9% and reduced the NIH-CPSI score by 24.4%. Combination of Qianlieping and tamsulosin more significantly increased the number of SPL (61.4%) and decreased the NIH-CPSI score (42.3%) than tamsulosin alone (33.7% and 28.6%) (P < 0.01).</p><p><b>CONCLUSION</b>Qianlieping Capsule chronic is effective for chronic non-bacterial prostatitis, and the combination of Qianlieping Capsule with tamsulosin produces even better effect than tamsulosin alone.</p>

Characteristics and risk factors of lymph node metastases in esophageal carcinoma / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the status of lymph node metastases (LNM) of esophageal carcinoma and to identify the risk factors.</p><p><b>METHODS</b>Clinical data of 308 patients who underwent esophagectomy with three-field lymphadenectomy during January 2006 and December 2010 were reviewed. Characteristics of LNM were studied.</p><p><b>RESULTS</b>The average number of dissected lymph nodes was 35.6 ± 14.5 in 308 patients. There were 197 patients(64%) had LNM. Logistic regression analysis showed that lymphatic vessel invasion(P=0.019) and deep tumor invasion(P<0.001) were risk factors of LNM. The highest LNM site was paratracheal node(25.0%). The incidence of cervical LNM was 14.1% in the middle thoracic carcinoma, higher than that of upper thoracic (7.3%) and lower thoracic (8.3%). Rate of LNM was lower in upper thoracic carcinomas than that in middle or lower ones(P=0.001). No significant difference of LNM was found among upper, middle and lower thoracic carcinoma for cervical or thoracic nodes. Lymphatic vessel invasion(P<0.001) and metastases in paratracheal lymph nodes (P=0.014) were risk factors for cervical LNM.</p><p><b>CONCLUSIONS</b>LNM of esophageal carcinoma can be found in both directions vertically and skipped metastasis. Paratracheal lymph nodes involvement is an indicator for cervical lymphadenectomy in thoracic esophageal carcinoma.</p>

Effects of prepubertal exposure to diethylstilbestrol on testicular development and function of SD rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of prepubertal exposure to diethylstilbestrol (DES) on the testicular development and function of Sprague-Dawley (SD) rats.</p><p><b>METHODS</b>Ninety 21-day-old male SD rats were randomly and equally divided into 4 experimental groups (Da, Db, Dc and Dd), which were injected with DES dissolved in corn oil at the dose of 0.01, 0.1, 1.0 and 10.0 microg/(kg x d) from postnatal day (PND) 22 to 35, and a control group (C), which received vehicle only. The testicular development of all the rats was observed, and their testes were harvested in the stages of late puberty (PND 50), sexual maturity (PND 64) and adulthood (PND 130) respectively to determine the weight and histological features of the testis and examine the quality of the sperm in the epididymal cauda of the PND 130 rats.</p><p><b>RESULTS</b>The testis descent in the C, Da, Db, Dc and Dd groups occurred on PND 26.17 +/- 1.94, 26.83 +/- 1.47, 28.68 +/- 1.03, 33.50 +/- 1.87 and 41.50 +/- 2.74 respectively, significantly delayed in the Db, Dc and Dd groups compared with the C group (P < 0.05 or P < 0.01). On PND 50, the unilateral testis weights in the C, Da, Db, Dc and Dd groups were (1.38 +/- 0.01) g, (1.38 +/- 0.12) g, (1.30 +/- 0.14) g, (0.86 +/- 0.18) g and (0.73 +/- 0.27) g respectively, significantly less in the Dc and Dd groups than in the C group (P < 0.01). Compared with the C group, there was a slight decrease in the number of the cells in the epithelia of a few seminiferous tubules in the Db group on PND 50, maldevelopment of seminiferous tubules, reduced cell number in seminiferous epithelia, blocked spermatogenesis and aplasia of Leydig cells in the Dc and Dd groups in a dose-dependent manner. On PND 64, the unilateral testis weights in the C, Da, Db, Dc and Dd groups were (1.60 +/- 0. 06) g, (1.62 +/- 0.11) g, (1.58 +/- 0.08) g, (1.47 +/- 0.10) g and (0.99 +/- 0.37) g respectively, significantly less in the Dc and Dd groups than in the C group (P < 0.05 or P < 0.01), and the histological alteration of the testis in the Dc and Dd groups was similar to or less than that on PND 50. On PND 130, no statistic difference was observed either in unilateral testis weight or in the histological features of the testis between any experimental group and the control (P > 0.05). The sperm concentration in the epididymal cauda in the C, Da, Db, Dc and Dd groups were (73.00 +/- 16.90) x 10(6)/ml, (68.00 +/- 19.67) x 10(6)/ml, (68.67 +/- 12.15) x 10(6)/ml, (35.17 +/- 15.64) x 10(6)/ml and (19.13 +/- 5.17) x 10(6)/ml, significantly lower in the Dc and Dd groups than in the C group (P < 0.01). There was a significant decrease in sperm motility in the Dd group (P < 0.01), the percentage of grade a sperm in the Db, Dc and Dd groups (P < 0.05) and the percentage of grade b sperm in the Dd group (P < 0.01).</p><p><b>CONCLUSION</b>Prepubertal exposure to low dose of DES (0.01 microg/[kg x d] x 14 d) does not significantly affect the testicular development and function of SD rats, while high dose (1.0-10.0 microg/[kg x d] x 14 d) has significant short- (PND 50 and 64) or long-term (PND 130) toxic effect, which increases with dose and decreases with age. The mechanism of the toxic effect involves the insults to the development and function of Leydig and Sertoli cells.</p>

Correlation of the prognosis of chronic prostatitis/chronic pelvic pain syndrome with psychological and other factors: a Cox regression analysis / 中华男科学杂志

<p><b>OBJECTIVE</b>To establish the role of psychological factors in the etiology and symptomatology of chronic prostatitis / chronic pelvic pain syndrome (CP/CPPS), analyze the influence of the psychological obstacles and other relative factors on the prognosis of CP/CPPS by univariate and multivariate Cox regression analyses, and provide a scientific basis for psychotherapy of the problem.</p><p><b>METHODS</b>A total of 291 CP/CPPS patients and 100 normal controls were investigated in age, education, occupation, character, disease course, NIH chronic prostatitis syndrome index (NIH-CPSI) and leukocyte count in EPS and by self-rating anxiety scale (SAS) and self-rating depression scale (SDS) to establish the psychological factors related with CP/CPPS. Then, all the CP/CPPS patients were treated with the same method and followed up for 6 weeks. Based on the therapeutic results, the influence of psychological and other relative factors on the prognosis of CP/CPPS was analyzed with univariate and multivariate Cox regression.</p><p><b>RESULTS</b>All together 258 valid questionnaires were collected from the patients and 87 from the normal controls. Of the 258 CP/CPPS patients, the mean scores on SAS and SDS were 42.8 +/- 11.43 and 48.15 +/- 11.49 respectively, both significantly higher than those of the controls (32.12 +/- 9.68 and 35.12 +/- 10.81) (P < 0.01). The rates of anxiety, depression and anxiety and/or depression in the CP/CPPS group were 25.97, 21.71 and 34.50 % respectively, all significantly higher than in the control group (P < 0.01). The rate of introversion was significantly higher while that of extroversion significantly lower in the former than in the latter (P < 0.01). The total effectiveness rate of treatment was 70.54 % in the CP/CPPS patients. Univariate and multivariate analyses with Cox regression revealed that anxiety, depression and disease course were the definite factors that negatively affected the prognosis of CP/CPPS, while the other factors, such as age, CPSI, character and leukocyte count in EPS had no influence.</p><p><b>CONCLUSION</b>Such psychological obstacles as anxiety and depression play an important role in the pathogenesis, development and prognosis of CP/CPPS. In the treatment of CP/ CPPS, importance should be attached to the patients'psychological status and proper psychological intervention is sometimes necessary.</p>

Clinicopathological significance of aromatase expression in breast cancers / 中华外科杂志

<p><b>OBJECTIVE</b>To study the effects of aromatase on breast cancer proliferation and invasive ability, so as to detect the relationship between in situ estrogen levels and molecular biological characteristics of breast cancer.</p><p><b>METHODS</b>By immunohistochemistry staining, the expression of aromatase, matrix metalloproteinases 2 (MMP2) and matrix metalloproteinases (MMP 9) in the primary breast cancers were detected, the associations between aromatase and MMPs as well as clinical-pathological factors were analyzed.</p><p><b>RESULTS</b>The positive rates of aromatase were 25.0% (+) and 29.9% (++). Aromatase status was associated with MMP2, MMP9 and co-expression of MMP2 and MMP9 (P < 0.05), but not associated with tumor size, ER/PR status, menopausal status and tumor grade (P > 0.05). In the postmenopausal patients there was a relationship between aromatase and tumor size (P < 0.05), but not in the premenopausal patients (P > 0.05); there was a relationship between aromatase and co-expression of MMP2/MMP9 in the patients with ER and/or PR positive (P < 0.05), but not in the patients with ER and PR negative (P > 0.05).</p><p><b>CONCLUSIONS</b>In the breast cancer in situ estrogen produced by tumor aromatase may promote the cancer cells proliferation and invasiveness and maybe through ER pathway especially in the postmenopausal patients.</p>

A preliminary study on the effect of prepubertal exposure of male rats to diethylstilbestrol on the apoptosis of spermatogenic cells after sexual maturation and its mechanism / 中华男科学杂志

<p><b>OBJECTIVE</b>To preliminarily study the effect of prepubertal exposure of male SD (Sprague-Dawley) rats to diethylstilbestrol (DES) on the apoptosis of spermatogenic cells after sexual maturation and its mechanism.</p><p><b>METHODS</b>Thirty 21-day-old male SD rats were randomly divided into 4 experimental groups, DES 0.01, 0.1, 1.0 and 10.0 microg/(kg x d) and 1 control group. The experimental groups were injected (s.c.) with different doses of DES (dissolved in corn oil) during prepuberty [from postnatal day (PND) 22 to PND 35] and the control group with medium only. The apoptosis and related proteins Bcl-2 and Bax expressions of testicular spermatogenic cells were studied with TUNEL and immunohistochemistry after the rats sexual maturation (at PND 64).</p><p><b>RESULTS</b>Compared with the control group, the apoptosis of testicular spermatogenic cells in the DES 0.01 microg/kg group had no difference, but significantly increased in the DES 0.1, 1.0 and 10.0 microg/kg groups and the apoptosis increased with the increase of DES dose. In the control and DES 0.01 microg/kg groups, Bax protein expressed weakly but Bcl-2 protein strongly in spermatogenic cells. With the increase of DES exposure, Bax protein expression in spermatogenic cells increased but Bcl-2 protein expression decreased.</p><p><b>CONCLUSION</b>Prepubertal exposure of SD rats to inappropriate dose of DES can make the apoptosis of spermatogenic cells increase after sexual maturation. Bax and Bcl-2 proteins participate in the apoptotic course caused by prepubertal DES exposure.</p>

In vitro study of the effects of estrogen receptor beta expression on the biological behavior of a human breast cancer cell line / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study effects of estrogen receptor beta (ER beta) on the biological behavior of a human breast cancer cell line MDA-MB-435.</p><p><b>METHODS</b>Human ER beta cDNA was introduced into MDA-MB-435 cells by stable transfection. Effects of ER beta expression on cell proliferation and invasion were investigated by MTT, flow cytometry and transwell techniques. Cyclin A, cyclin E, cyclin D1, p21, MMPs, Ets-1, VEGF and b-FGF were detected by RT-PCR and/or Western blot or gelatin zymography.</p><p><b>RESULTS</b>ER beta was shown to be able to significantly increase the proliferation and invasion of MDA-MB-435 cells in an estradiol-independent manner. The S phase distribution of the cells with ER beta overexpression was 46.8%, significantly higher than that of wild type (29.9%) and mock transfected cells (27.6%) (P = 0.01). In ER beta transfected cells, the expression of p21 decreased by 33.3% at mRNA level (P = 0.03) and by 47.4% at protein level (P = 0.02), respectively. The expression of MMP-9 increased by 91.3% at mRNA level (P < 0.01) and its activity was up-regulated by 67.3% (P = 0.02). Furthermore, the mRNA and protein levels of Ets-1 increased 62.2% (P = 0.01) and 51.0% (P = 0.01), respectively. No significant difference was observed in the mRNA levels of cyclin A, cyclin E, cyclin D1, MMP-1, MMP-2, MMP-7, VEGF and b-FGF among these cells.</p><p><b>CONCLUSION</b>ER beta can enhance proliferation and invasion of breast cancer cells. Down-regulation of p21 and up-regulation of MMP-9 and Ets-1 may be involved in its mechanisms.</p>

Pattern of lymph node metastasis in patieats with squamous cell carcinoma of the thoracic esophagus and its significance in lymphadenectomy / 中华胸心血管外科杂志

Objective To explore the pattern of lymph node metastasis in patitsen with squamous cell carcinoma of the thoracic esophagus and its significance in lymphadenectomy.Methods The clinical data of 230 patients who received radical esophagectomy with three-field lymphadenectomy was analyzed.The metastatic sites of lymph nodes were correlated with tumor location by chi-square test.Logistic regression was used to analyze the relationship between clinic pathoingical factors and lymph node metastasis.Results Lymph node metastases were found in 133 of the 230 patients(57.8%).The average number of resected lymph nodes was 25.3? 11.4 per patient(range 11～71).The rates of lymph node metastasis were 41.6%,19.44%and 8.3%in the neck,thoracic medi- astinum and abdominal cavity for patients with upper thoracic esophageal carcinoma,33.3%,34.7%and 14%for patients with mid- die thoracic esophageal carcinoma and 36.4 %,34.1%and 43.2 %for patients with lower thoracic esophageal carcinoma.No signifi- cant difference in cervical or thoracic metastatic rate was observed among upper,middle and lower thoracic carcinoma.The difference in lymph node metastatic rate for nodes in the abdominal cavity was significant among upper,middle and lower thoracic carcinoma. The lower thoracic esophageal cancers were more likely to metastasize to the abdominal cavity.Logistic-regression showed depth of tu- mor invasion and angiolymphatic invasion were factors influencing lymph node metastasis.Conclusion Cervical and mediastinal node dissection should be performed independently from tumor location.Abdominal node dissection should be conducted more vigorously for lower thoracic esophageal cancer than of other locations.Patients with greater tumor grade,depth of tumor invasion and angiolymphatic invasion were more prone to develop lymph node metastasis.

Inhibition effects of epigallocatechin-3-gallste (EGCG) on cell cycle of hum an breast cancer cells MDA-MB-435 and its mechanism / 中国癌症杂志

Purpose:To study the effect of EGCG(extracts fr om green tea) on cell cycle in human breast cancer cell line and its mechanism. Methods:Cell proliferation assay kit was used to produce the dose-response curve. Flow cytometric assay was used to evaluate the cell cycle changes on MDA-MB435 cells with and without EGCG. The expression of protein was detected by Western blot. Results:EGCG could inhibit the proliferation of the breast canc er cells MDA-MB-435, 40 ?g/ml EGCG induced G 0 /G 1 phase arrest and the entrance to S phase. Both mRNA and protein level of p21 waf1/cip1 were up regulated in MDA-MB435 cells when treated by 40?g/ml EGCG. Conclus ions:Green tea can inhibit the growth of human breast cancer cells, perhaps by means of p21 and therefore inhibiting the progression of the cell cycle.