Testosterone levels in patients with varicocele and azoospermia / 北京大学学报(医学版)

OBJECTIVE@#Androgen deficiency is common in aging males and may have unfavourable health consequences. Large-scale studies suggested low testosterone level might increse mortality and morbidity in ageing males. However, young men with low testosterone level might be neglected. Recent studies reported young men with infertility may have reduced testosterone level. To investigate the incidence of androgen deficiency in males with infertility and possible factors affecting the low testosterone level.@*METHODS@#Between January 2011 and December 2012, 407 men with infertility caused by varicocele (VC), obstructive azoospermia (OA) and nonobstructive azoospermia (NOA) in our center were included. The number of men in each group of OA, NOA and VC was 141, 97 and 169, respectively. All the eligible patients underwent a serum testosterone assessment by a single morning blood draw (between 8:00 to noon) to test for concentration of the total testosterone. All serum samples were determined by radioimmunoassay in our andrology laboratory. Androgen deficiency was defined as having a total testosterone level less than 300 ng/dL.@*RESULTS@#The mean age was (30.4±5.8) years. The mean testosterone level was (4.18±1.64) ng/dL (range 0.30 to 11.32 ng/dL). The overall incidence of androgen deficiency was 26.5% (108/407). The incidences of androgen deficiency in NOA, OA and VC groups were 40.2% (39/97), 19.1% (27/141) and 24.9% (42/169), respectively, which were significantly higher in the NOA than in the VC and OA groups (P < 0.001). The incidences had no difference between the VC and OA groups (P=0.229). Univariate analysis revealed the cause of infertility, FSH and the mean testis volume as possible affecting factors for androgen deficiency. However, on multivariate analysis the only cause of infertility was an independent predictor. The incidence of androgen deficiency was the highest in the NOA group [OR 0.492 (95% confidence interval 0.288-0.840)].@*CONCLUSION@#NOA and varicocele might be risk factors of androgen deficiency. Young men with NOA may have a higher possibility of low testosterone level. Testosterone level should be followed up after NOA and varicocele treatment. Androgen deficiency should be assessed in males with infertility in clinical practice.

Appllication of human acellular dermal matrix in surgical treatment of genitourinary disease / 北京大学学报(医学版)

Humanacellular dermal matrix (HADM) is widely used in the field of burn wound repair and tissue engineering plastic surgery. HADM is manufactored by physical and chemical decellular process to remove the antigenic components that might cause immune rejection in dermis.The extracellular matrix of three-dimensional cell scaffold structure with collagen fibers had been used for wound repair and tissue regeneration, while HADM characterized with low absorption rate after implantation and strong ability to induce angiogenesis in host tissue. Studies reported that after the HADM was implanted into the patient, the host cells, such as fibroblasts and myofibroblasts, as well as lymphocytes, macrophages, granulocytes and mast cells, rapidly infiltrated the graft. The connective tissue and neovascularization were then formed within the HADM three-dimensional cell scaffold, the lymphatic system also appears after vascular reconstruction. Traditional urethral reconstruction using autologous skin flaps has some defects, such as complexity of the technology, risk of necrosis of the skin flaps after transplantation, and failure to achieve functional repair of the urethral epithelium. It has been reported that using HADM to reconstruct the urethra in patients with urethral stricture, hypospadias and bladder-vaginal fistula, showed promising results. Others have reported the experience of using HADM to repair and reconstruct congenital classic bladder exstrophy. HADM has also been used for tissue repair in patients with penile skin defect caused by Fonier's gangrene and hidradenitis suppurativa, and implanted under Bucks' fascia to enlarge the penis. The report of HADM implantation for treating premature ejaculation also deserves attention. Researchers found that HADM implantation can form a tissue barrier between the skin and corpus cavernosum, which can effectively reduce penile sensitivity and treat premature ejaculation. The safety and effectiveness of HADM implantation in the treatment of premature ejaculation need to be further standardized by data from multi-center, large-sample clinical studies. In summary, HADM is the extracellular matrix and three-dimensional cell scaffold of human dermis. As a new type of tissue repair material, new blood vessels are formed actively after implantation, which shows good histocompatibility. HADM has shown increasingly broad application prospects in treatment of genitourinary diseases including penis, urethra and bladder diseases. HADM has also been used in the treatment of premature ejaculation in recent clinical studies, and its long-term safety and efficacy need to be further investigated.

Chloroquine inhibits viability of renal carcinoma cells and enhances sunitinib-induced caspase-dependent apoptosis / 北京大学学报(医学版)

OBJECTIVE@#To determine whether chloroquine (CQ), an often used inhibitor of late autophagy and autophagosome/lyosome fusion, can inhibit proliferation of renal carcinoma cells and investigate its effect on sunitinib (ST)-induced apoptosis.@*METHODS@#Renal carcinoma cell line 786 O and ACHN had been used as cellular model and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay was carried out to detect the cell viability in response to CQ or ST treatment. Both transmission electron microscope and immunoblotting had been employed to observe apoptotic and autophagic process. To examine the involvement of autophagy in ST-dependent apoptosis, autophagy had been inhibited either chemically or genetically via utilizing autophagy inhibitor or specific small interference RNA (siRNA) targeted to either Ulk1 (unc-51-like kinase 1) or LC3 (microtubule associated protein 1 light chain 3 fusion protein), two essential autophagic proteins.@*RESULTS@#Both ST and CQ induced cell viability loss, indicating that either of them could inhibit renal cancer cell proliferation. Clone formation experiments confirmed the aforementioned results. Furthermore, the combined ST with CQ synergistically promoted the loss of cell viability. By transmission electron microscopy and immunoblotting, we found that the ST induced both autophagy and caspase-dependent apoptosis. While 3-MA, an early autophagy inhibitor, reduced the ST-induced cleavage of poly (ADP-ribose) polymerase-1 (PARP-1), a substrate of caspase 3/7 and often used marker of caspase-dependent apoptosis, CQ promoted the ST-dependent PARP-1 cleavage, indicating that the early and late autophagy functioned differentially on the ST-activated apoptotic process. Moreover, the knock down of either Ulk1 or LC3 decreased the ST-caused apoptosis.Interestingly, we observed that rapamycin, a specific inhibitor of mTOR (mammalian target of rapamycin) and an inducer of autophagy, also showed to inhibit cell viability and increased the cleavage of PARP-1 in the ST-treated cells, suggesting that autophagy was likely to play a dual role in the regulation of the ST-induced apoptosis.@*CONCLUSION@#ST activates both apoptotic and autophagic process in renal carcinoma cells. Although autophagy precedes the ST-induced apoptosis, however, early and late autophagy functions differentially on the apoptotic process induced by this compound. Additionally, ST can coordinate with the inducer of autophagy to inhibit the cell proliferation. Further research in this direction will let us illuminate to utilize CQ as a potential drug in the treatment of renal carcinoma.

Effects of intensive therapy program on gross motor function of children with spastic cerebral palsy / 医用生物力学

Objective To explore the effects of intensive therapy program on gross motor function of children with spastic cerebral palsy. Methods Thirty 3-15 year old children with spastic cerebral palsy and level I-III in gross motor function classification system (GMFCS) were randomly divide into 2 groups. The trial group adopted intensive therapy program for treatment, including functional dynamic suit and universal exercise unit combined with functional movement training. The control group adopted core stability training for treatment. Children in two groups took a 1-month training program for 3 hours per day and 5 days per week. The pediatric evaluation of gross motor function measure (GMFM-66) and peabody developmental motor scale (PDMS-2) were administered before and after treatment. Results The scores of GMFM-66 as well as the stationary and locomotion scores of PDMS-2 in both groups showed significant differences within group (P<0.01) after treatment. For the scores of GMFM and PDMS-2 between two groups, no significant differences were found. Conclusions The intensive therapy program can improve the gross motor function of children with spastic cerebral palsy and gain the same effects as core stability training, which can provide a novel and effective intervention for children with cerebral palsy.

Effects of intensive therapy program on gross motor function of children with spastic cerebral palsy / 医用生物力学

Objective To explore the effects of intensive therapy program on gross motor function of children with spastic cerebral palsy.Methods Thirty 3-15 year old children with spastic cerebral palsy and level Ⅰ-Ⅲ in gross motor function classification system (GMFCS) were randomly divide into 2 groups.The trial group adopted intensive therapy program for treatment,including functional dynamic suit and universal exercise unit combined with functional movement training.The control group adopted core stability training for treatment.Children in two groups took a 1-month training program for 3 hours per day and 5 days per week.The pediatric evaluation of gross motor function measure (GMFM-66) and peabody developmental motor scale (PDMS-2) were administered before and after treatment.Results The scores of GMFM-66 as well as the stationary and locomotion scores of PDMS-2 in both groups showed significant differences within group (P ＜ 0.01) after treatment.For the scores of GMFM and PDMS-2 between two groups,no significant differences were found.Conclusions The intensive therapy program can improve the gross motor function of children with spastic cerebral palsy and gain the same effects as core stability training,which can provide a novel and effective intervention for children with cerebral palsy.

Daily low-dose tadalafil for erectile dysfunction induced by pelvic fracture urethral disruption / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of daily low-dose tadalafil on erectile dysfunction (ED) induced by pelvic fracture urethral disruption (PFUD).</p><p><b>METHODS</b>This study included 46 cases of PFUD-induced ED treated from Jan 2008 to Dec 2011. The patients were aged 33.9 +/- 7.2 years (range 25 -51 yr), and the interval between injury and treatment was 19.6 +/- 12.7 months (range 3 - 72 mo), all with normal erectile function before PFUD. Based on the nocturnal penile tumescence and rigidity (NPTR) recorded by erectometry without medication of phosphodiesterase type 5 inhibitor (PDE-5I), the patients were divided into an abnormal nocturnal erection group and a non-nocturnal erection group, and treated with tadalafil 10 mg once daily for 3 months. The therapeutic effect was evaluated by IIEF-5 scores and the rate of yes responses to questions 2 and 3 of the Sexual Encounter Profile (SEP).</p><p><b>RESULTS</b>Totally 38 (82.6%) of the patients accomplished the treatment and follow-up, 26 (68.4%) in the abnormal nocturnal erection group and 12 (31.6%) in the non-nocturnal erection group. After 3 months of daily tadalafil treatment at 10 mg, the IIEF-5 scores were significantly improved in the abnormal nocturnal erection group than in the non-nocturnal erection group (P < 0.05), and the rate of yes responses to SEP2 and SEP3 was remarkably higher in the former than in the latter (76.9% vs 41.7% and 65.4% vs 25.0%, P < 0.05).</p><p><b>CONCLUSIONS</b>Daily low-dose tadalafil can effectively improve the erectile function of PFUD-induced ED patients, particularly in those with nocturnal erection.</p>

Expression and localization of CKLFSF2 in human spermatogenesis / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To investigate the expression and subcellular localization of chemokine-like factor superfamily 2 (CKLFSF2) in human testis and its potential role in spermatogenesis.</p><p><b>METHODS</b>A specific polyclonal antibody against CKLFSF2 was raised. The expression and cellular localization of CKLFSF2 in the seminiferous tubules was checked by immunohistochemistry method. Also, in situ hybridization was applied to localize the mRNA distribution. The EGFP-CKLFSF2 fusion protein was expressed in COS-7 cells to localize its subcellular location in vitro. In addition, the abnormal expression of CKLFSF2 in testes of patients with male infertility was assayed by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry methods.</p><p><b>RESULTS</b>Having a close correlation with spermatogenesis defects, CKLFSF2 was specifically expressed in meiotic and post-meiotic germ cells, which were localized to the endoplasmic reticulum (ER) near the Golgi apparatus.</p><p><b>CONCLUSION</b>CKLFSF2 could play important roles in the process of meiosis and spermiogenesis, and might be involved in the vesicular transport or membrane apposition events in the endoplasmic reticulum.</p>

Construction and expression of ATP50 fluorescent protein in TM3 mouse Leydig cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the expression and localization of ATP50 by construction of ATP50-pEYFP-N1 in primary cultured mouse Leydig cells.</p><p><b>METHODS</b>Primary cultured mouse Leydig cells were confirmed by 3B-HSD staining. ATP50 was cloned into pEYFP-N1 between Bam HI and Eco RI sites. Cell-transfection and living-cell fluorescence imaging microscopy were employed to investigate the sub-cellular localization of YFP-ATP50 in TM3 mouse Leydig cells.</p><p><b>RESULTS</b>ATP50 green fluorescent protein was well co-localized with red fluorescence mitochondrion marker-Mitotracker in TM3 mouse Leydig cells.</p><p><b>CONCLUSION</b>ATP50 was expressed in primary cultured mouse Leydig cells. The fluorescent expression vector of ATP50 was constructed successfully and YFP-ATP50 was located in mitochondria in TM3 mouse Leydig cells, which provided a useful clue for further research on the steroidogenesis dysfunction in aging males.</p>

Construction of pGEX4T-1-Cox7a2 and expression, purification and identification of the recombinant protein / 中华男科学杂志

<p><b>OBJECTIVE</b>To clone and express Cox7a2, one mitochondrial respiratory chain related gene, and to identify its recombinant protein.</p><p><b>METHODS</b>The coding region of Cox7a2 was amplified from primary cultured mouse Leydig cells by RT-PCR. The PCR product was cloned into pGEX4T-1 vector by BamH I and EcoR I sites, and confirmed by DNA sequencing. The recombinant fusion protein vector was transformed and expressed into BL21. The recombinant fusion protein was identified by Western blotting.</p><p><b>RESULTS</b>The entire coding region of Cox7a2 was cloned and expressed. The fusion protein was identified by anti-GST monoclonal antibody using Western blotting.</p><p><b>CONCLUSION</b>The cloning of Cox7a2 and the expression of the recombinant protein would help to study the detailed function of Cox7a2, one respiratory chain related and highly differently expressed gene in the tissues of aging testes.</p>

Influence of bi-level positive airway pression ventilation via nasal on ET-1 of overlap syndrome / 中国基层医药

Objective To explore the effect of nBiPAP on ET-1 of patients with overlap syndrome.Methods Levels of ET-1 in plasma of 25 cases of OS,20 cases of COPD were analysed by radioimmunoassay,and ET-1 in OS was measured before and after treatment.The relationship between ET-1 with SaO_2 during sleep was analyzed. Results (1)The ET-1 levels in plasma of OS group were significantly higher than those of COPD group before treatment(P