Condom stigma among men who have sex with men population: Concept synthesis / 中南大学学报(医学版)

OBJECTIVES@#The epidemic of acquired immune deficiency syndrome (AIDS) among men who have sex with men (MSM) is severe in China. And MSM has now become a key population for the infection and transmission of AIDS. At present, the bottleneck of AIDS prevention and control among MSM population is low rate of continuous condom use and high incidence of unsafe sexual behavior. Inductive summarization of the literature revealed that the most critical reason for low rate of continuous condom use among the MSM population was condom-related stigma. Although many studies mentioned condom-related stigma among MSM populations, there has been no any definition of MSM-related condom stigma and no measurement for it. Therefore, the paper aims to explore barriers to condom use among MSM, then construct the conceptual and operational definition of "MSM-related condom stigma" through Meta synthesis and concept synthesis, and provide a new perspective for AIDS prevention and control among MSM.@*METHODS@#Based on evidence-based method, "PICoS" framework and Meta-synthesis was used to include the literatures. Then, we used synthesized qualitative evidence from included studies to construct the concept and operational definition of MSM-related condom stigma by the means of thematic analysis and concept synthesis.@*RESULTS@#According to the results of the concept synthesis, MSM-related condom stigma refers to any taboos or misbeliefs about condom use or feeling ashamed or embarrassed to talk about using condoms which perceived by individuals at the individual, interpersonal, and social levels.It was demonstrated through 4 sub-themes at operational level: a symbol of distrust, a symbol of HIV/sexual transmitted infections (STIs) prevention, a symbol of an embarrassing topic, and a symbol of violating the traditional cognition of sexual intercourse. According to the Social-ecological Model (SEM), a symbol of distrust refers to that the MSM population believes that not using condoms represents mutual trust between sexual partners, while using condoms is difficult to express intimacy, trust and loyalty between sexual partners. A symbol of HIV/STIs prevention at the interpersonal level refers to that the MSM population believes that condom use is a "symbol" for the prevention or infection of AIDS; on the one hand, if someone proposes to use condoms, he may be considered infected with HIV or have unsafe sex experiences, thus, making it difficult to propose condom use; on the other hand, if they believe that sexual partners are "AIDS free" (often a wrong perception, such as sexual partners may have the risk of AIDS infection although they do not have AIDS), it is considered that condom use is completely unnecessary. The environmental level includes a symbol of an embarrassing topic and a symbol of violating the traditional cognition of sexual intercourse. A symbol of an embarrassing topic refers to the MSM population feels shame about topics related to sexual behavior and is embarrassed to carry/buy/propose condom use or be ashamed to engage in conversations about whether to use condoms during sexual behavior. And a symbol of violating the traditional cognition of sexual intercourse: The MSM population have limitations in their perception of "sex" or "sexual behavior" and believe that real sex (behavior) is unobstructed contact between the bodies and exchange between all body fluids.@*CONCLUSIONS@#The concept of MSM-related condom stigma is proposed for the first time, and its operational definition is given. The concept includes 3 levels and 4 dimensions. It is helpful to understand MSM people's attitude and cognition towards condoms, and adds indicators with cultural sensitivity and behavioral sensitivity to the behavioral intervention for AIDS in the future.

Effect of Mobile Health for Standardized Management on Women with Gestational Diabetes Mellitus / 中国医学科学院学报

Objective To explore the performance of mobile health platform for standardized management of pregnant women with gestational diabetes mellitus(GDM). Methods A randomized controlled trial was conducted,in which 295 women with GDM were randomized into two groups(traditional management group and mobile health management group)by a computer-generated sequence.The traditional management group accepted standardized GDM management,and the mobile health management group was supplemented by mobile health management based on the standardized management.The glycemic control rate and the incidences of low birth weight,macrosomia,preterm birth,premature rupture of membranes,postpartum hemorrhage after cesarean section,neonatal asphyxia,malformation,and admission to the neonatal intensive care unit were compared between the two groups. Results The glycemic control rate in mobile health management group was significantly higher than that in the traditional management group [(67.22±22.76)%

Effect of active component compound of Epimedii Folium,Astragali Radix,and Puerariae Lobatae Radix on expression of ADAM17 in HT22 cells by mediating hepcidin / 中国中药杂志

Alzheimer's disease(AD) patients in China have been surging, and the resultant medical burden and care demand have a huge impact on the development of individuals, families, and the society. The active component compound of Epimedii Folium, Astragali Radix, and Puerariae Lobatae Radix(YHG) can regulate the expression of iron metabolism-related proteins to inhibit brain iron overload and relieve hypofunction of central nervous system in AD patients. Hepcidin is an important target regulating iron metabolism. This study investigated the effect of YHG on the expression of a disintegrin and metalloprotease-17(ADAM17), a key enzyme in the hydrolysis of β amyloid precursor protein(APP) in HT22 cells, by mediating hepcidin. To be specific, HT22 cells were cultured in vitro, followed by liposome-mediated siRNA transfection to silence the expression of hepcidin. Real-time PCR and Western blot were performed to examine the silencing result and the effect of YHG on hepcidin in AD cell model. HT22 cells were randomized into 7 groups: control group, Aβ25-35 induction(Aβ) group, hepcidin-siRNA(siRNA) group, Aβ25-35 + hepcidin-siRNA(Aβ + siRNA) group, Aβ25-35+YHG(Aβ+YHG) group, hepcidin-siRNA+YHG(siRNA+YHG) group, Aβ25-35+hepcidin-siRNA+YHG(Aβ+siRNA+YHG) group. The expression of ADAM17 mRNA in cells was detected by real-time PCR, and the expression of ADAM17 protein by immunofluorescence and Western blot. Immunofluorescence showed that the ADAM17 protein expression was lower in the Aβ group, siRNA group, and Aβ+siRNA group than in the control group(P<0.05) and the expression was lower in the Aβ+siRNA group(P<0.05) and higher in the Aβ+YHG group(P<0.05) than in the Aβ group. Moreover, the ADAM17 protein expression was lower in the Aβ+siRNA group(P<0.05) and higher in the siRNA+YHG group(P< 0.05) than in the siRNA group. The expression was higher in the Aβ+siRNA+YHG group than in the Aβ+siRNA group(P<0.05). The results of Western blot and real-time PCR were consistent with those of immunofluorescence. The experiment showed that YHG induced hepcidin to up-regulate the expression of ADAM17 in AD cell model and promote the activation of non-starch metabolic pathways, which might be the internal mechanism of YHG in preventing and treating AD.

Correlation analysis of prostate biopsy and radical prostatectomy specimen / 临床外科杂志

Objective To study the reliability of Gleason score by prostate biopsy with prostatic cancer(PCa)in prediction of tumor location and analyse related influencing factors of positive surgical margins after radical prostatectomy.Methods The retrospective study recruited 72 patients with PCa who had been diagnosed by trans rectal ultrasound-guided prostate biopsy,and eventually treated with laparoscopic radical prostatectomy.The correlation of each index were analyzed by statistical software.Results In this study,Kappa test indicated a good agreement that Gleason score between prostate biopsy and radical prostatectomy.Biopsy positive more focused on four peripheral points near the bottom.Biopsy tumor location was not suitable for predicting tumor distribution.Biopsy Gleason score and biopsy positive percentage in biopsy specimen were independent predictors of positive surgical margins.F and L points had good correlation with the positive surgical margins.Conclusion The biopsy Gleason score is a good evidence to pathological grading in patients.These information will lead to positive surgical margins rise possibly:needle biopsy Gleason score exceed 7.25,positive biopsy percentage more than 45% and the location positive biopsy close to prostate apex.

The history of prostate biopsy and recent research situation / 中国医师进修杂志

According to the latest data from the National Cancer Center of China, prostate cancer (PCa) has become the highest incidence tumors of urinary since 2008, and its incidence and mortality has occupied great attention in the past decades. Therefore, the choice of safe and effective diagnostic method, to detect the occurrence of PCa, is necessary. The way of clinical diagnosis mainly includes digital rectal examination, serum PSA, transrectal ultrasound, MRI spectroscopy imaging, and prostate biopsy, ect. Prostate biopsy is the most reliable diagnosis method and agold standardfor the diagnosis of PCa. The systematic use of the prostate biopsy has greatly improved the diagnosis of PCa. However, there are several prostate biopsy scheme and scholars proposed a variety of programs on the choice of puncture point. It has not yet formed a standard method at this stage. In this study, we will review the development of prostate biopsy and the status recent research to explore application value of different method.

Advances in tubeless percutaneous neprolithotomy / 中国综合临床

Percutaneous neprolithotomy (PCNL) is the preferred method for the treatment of the majority of urinary calculi, especially for the calculi lies in the renal and upper ureteral.Recently, a number of trials were applied into TPCNL, such as, decrease the nephrostomy and indwelling externally grooved ureteral stent, internal ureteral catheter and not aiming replace the preventive use of renal tube.TCNL has its advantages in postoperative pain, hospitalization time and postoperative recovery than standard percutaneous neprolithotomy.

Prostatic arterial embolization for benign prostatic hyperplasia in high-risk aged males / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the efficacy and safety of prostatic arterial embolization (PAE) in the treatment of benign prostatic hyperplasia ( BPH) in high-risk aged males.</p><p><b>METHODS</b>We retrospectively analyzed the clinical data about 21 high-risk BPH patients aged 77-91 (mean 80) years treated by PAE.</p><p><b>RESULTS</b>PAE was successfully performed in all the 21 patients, with the operation time of 90-120 min. At 2 weeks, 3 months, 6 months, and 12 months after surgery, the International Prostate Symptom Scores (IPSS) were 18.3 ± 3.1, 9.8 ± 2.7, 9.4 ± 2.5, and 10.1 ± 2.2, the quality of life scores ( QOL) were 4.6 ± 1.4, 4.3 ± 1.2, 4.6 ± 1.1, and 4.9 ± 0.6, the maximum urinary flow rates ( Qmax) were (12.5 ± 2.5), (15.8 ± 2.4), (16.6 ± 2.2), and (16.3 ± 1.8) ml/s, and the postvoid residual urine volumes (PVR) were (35.0 ± 3.4), (13.0 ± 3.3), (10.0 ± 3.0), and (8.0 ± 2.5) ml, respectively, markedly improved as compared with the baseline (IPSS: 24.5 ± 3.7, QOL: 5.7 ± 1.6, Qmax: [8.3 ± 2.1] ml/s, and PVR: [98.0 ± 11.0] ml), with statistically significant differences in IPSS, QOL, Qmax, and PVR (all P < 0.05). The maximal velocity of blood flow in the prostate was obviously decreased and the prostate volumes were (74.4 ± 4.8), (42.5 ± 4.4), (38.3 ± 4.0), and (36.7 ± 3.5) cm3 at 2 weeks, 3 months, 6 months, and 12 months, respectively, also significantly reduced in comparison with (84.3 ± 5.4) cm3 preoperatively (all P < 0.05).</p><p><b>CONCLUSION</b>PAE is a safe and effective option for the treatment of BPH in high-risk aged males.</p>

Comparation of efficacy between percutaneous decortication and ureteroscopic decompression in treating simple renal cyst / 国际外科学杂志

Objective To compare the efficacy and complications of percutaneous nephroscope decortication of cystic renal disease with transureteroscopic decompression of cystic renal disease.Methods Retrospectively analyze the clinical data of 42 simple renal cyst cases,who under treatment of surgical in Zhongnan Hospital of Wuhan University from Sep.2010 to Sep.2014 via percutaneous nephrolithotomy as well as ureteroscope.There were 21 patients in each group.Comparisons were made between the two groups on operation time,peripheral tissue injury,blood loss,postoperative infection,hospitalzation time.Postoperative recurrence were followed up.Results For the percutaneous nephroscope decortication of cystic renal disease group and transureteroscopic decompression of cystic renal disease group,the operation time were (38.43 ± 9.76) minutes,(28.95 ± 8.67) minutes,the number of tissue injury were 8,6;the blood loss were (28.62 ± 9.82) mL,(23.48 ± 7.65) mL;the number of postoperative infection was 4,10;the postoperative recurrence were 2,5;the hospitalzation time were 2 days and 8 days.Compared with the transureteroscopic decompression group,the percutaneous nephroscope decortication group had a less postoperative infection and fewer postoperative recurrence (P ＜ 0.05).But the operation time was longer in the percutaneous nephroscope decortication group (P ＜ 0.05).Conclusions The therapeutic effect of percutaneous nephroscope decortication is much better than that of transureteroscopic decompression,but also has a little disadvantage.

Combined subfascial endoscopic perforator surgery and endovenous laser treatment without impact on the great saphenous vein for management of lower-extremity varicose veins / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Conventional high ligation and stripping of the great saphenous vein (GSV) has a good curative effect but is highly traumatic with a considerable relapse rate. Subfascial endoscopic perforator surgery (SEPS) plus endovenous laser treatment (EVLT) could be applied as individual therapy. This study aimed to evaluate the feasibility of performing combined SEPS and EVLT without impacting GSV in the management of valvular insufficiency of the lower-limb venous perforators.</p><p><b>METHODS</b>Placement of lower-limb venous perforator insufficiency was marked by ascending phlebography in 83 affected limbs from September 2010 to June 2011. After randomization, SEPS was performed on 41 limbs to address the insufficiency of the venous perforators under the deep fascia, in combination with EVLT to close the superficial varicose veins without impacting the GSV. The remaining 42 limbs were treated using traditional GSV phlebectomy as controls.</p><p><b>RESULTS</b>Postoperatively, all varicose veins were resolved, with lightening of the pigmentation and healing of the ulcer. Within a follow-up period of 5 - 11 months, no symptoms had recurred. Compared with the control group, the operation time, the number of incisions sutured, and the in-hospital time decreased on average by 1.5 hours, 4.7, and 6.8 days, respectively (P < 0.01 in all cases).</p><p><b>CONCLUSION</b>Combined SEPS and EVLT for treatment of valvular insufficiency of the lower-limb venous perforators offer the advantages of microtrauma and rapid cure.</p>

Establishment of cryptorchid infertile models by postnatal injection of estradiol in mice / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the incidence and spermatogenesis of cryptorchid testes induced by postnatal injection of estradiol.</p><p><b>METHODS</b>Ninety male newborn Balb/C mice were randomly divided into an experimental (n = 60), a solvent control (n = 20) and a normal control group (n = 10). The experimental mice were again assigned to a 4-week, a 6-week, an 8-week, and a 10-week subgroup, and injected subcutaneously with 17-beta estradiol (5 microg/d) from 3 to 28, 3 to 42, 3 to 56 and 3 to 70 days after birth, respectively. The incidence of cryptorchidism and morphological changes of the testes were observed at 2 weeks after drug withdrawal.</p><p><b>RESULTS</b>The incidence rates of cryptorchidism in the 4-, 6-, 8- and 10-week groups were 0%, 26.7%, 60% and 60%, respectively, but no cryptorchidism occurred in the solvent and normal control groups. The 4- and 6-week groups showed autonomous descent of the cryptorchid testes and recovery of spermatogenesis after drug withdrawal. The models became stable and no spermatogenesis recovery was observed after 8 weeks of continuous medication.</p><p><b>CONCLUSION</b>Stable cryptorchid infertile models can be established in mice by postnatally continuous injection of estradiol for over 8 weeks.</p>

Expression of insulin-like factor 3 in the testis of flutamide-induced cryptorchidism mice and its significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the changes in the mRNA expression of insulin-like factor 3 (INSL-3) in the testis of mouse models of flutamide-induced cryptorchidism.</p><p><b>METHODS</b>We randomized pregnant BALB/c mice to groups A (control) , B, C, D and E to receive continuous gavage of flutamide at 0, 150, 300, 500 and 700 mg/kg body weight, respectively, from gestation day 12 to 21. We detected the expression of INSL-3 mRNA in the testis of the neonates by real-time PCR at 4 and 8 postnatal weeks.</p><p><b>RESULTS</b>No cryptorchidism was found in group A; unilateral cryptorchidism was seen in groups B (10.0%) and C (25.0%); and bilateral cryptorchidism was observed in groups D (21.1%) and E (40.0%). The expression of INSL-3 mRNA was reduced with the increased dose of flutamide, not significantly changed in groups B and C (P > 0.05) but remarkably decreased in D and E as compared with A (P < 0.05).</p><p><b>CONCLUSION</b>Administration of flutamide to pregnant mice can induce unilateral cryptorchidism at 150 and 300 mg/kg and bilateral cryptorchidism at 500 and 700 mg/kg in their male offspring, which may be related with its reducing effect on the expression of INSL-3 in the testis of the mice.</p>

Characteristics of patients with fluid extravasation during retrograde ureteroscopic holmium laser lithotripsy for renal calculi

To explore the risk factors of fluid extravasation during retrograde ureteroscopic holmium laser lithotripsy for renal calculi. Three hundred and twenty-seven patients with renal calculi ranging 10 to 20 mm received retrograde ureteroscopic holmium laser lithotripsy at Renmin Hospital of Wuhan University, Wuhan, China from January 2004 to December 2010. The clinical records were reviewed, and the correlation was studied between various clinical factors and fluid extravasation complications during operation. The clinical factors to be tested included patients' gender and age [<30, 30-50, and >50 years], hydronephrosis degree, previous intervention for renal calculi [none, shock-wave lithotripsy, and open surgery], upper urinary tract infection, ureteral access sheath placement, and procedure duration [<60, 60-120, and >120 mins]. The data were processed by SPSS Version 16.0 statistical software, x[2] test, and binary logistic regression were used for analysis. Fluid extravasation complications appeared in 35 patients. Patients` gender, age, and hydronephrosis degree were irrelevant to the occurrence of fluid extravasation, while having previous open surgery for renal calculi, without ureteral access sheath placement, upper urinary tract infection, and long procedure duration were all responsible for higher incidence of the complications. Reasonable selection of patients, effective control of upper urinary tract infection, routine ureteral access sheath placement, and controlling procedure duration help to decrease the incidence of fluid extravasation complications in retrograde ureteroscopic holmium laser lithotripsy for renal calculi

Murine model of busulfan-induced spermatogenesis regeneration: a quantitative evaluation / 中华男科学杂志

<p><b>OBJECTIVE</b>To quantitatively evaluate the murine model of spermatogenesis regeneration induced by two-dose busulfan injection.</p><p><b>METHODS</b>Fifty-four male mice were randomly divided into a control and two model groups of equal number, the former treated by two-dose intraperitoneal injection of 50% DMSO solution at 10 ml/kg, and the latter by that of busulfan at 10 mg/kg and 15 mg/kg respectively to establish spermatogenesis regeneration models, both at the interval of 24 days between the two doses. Spermatogenesis in seminiferous epithelia was evaluated by Johnsen score, and the expressions of GATA-4 and GDNF mRNA in Sertoli cells were detected by real time quantitative PCR at 3, 4 and 8 weeks after the treatment.</p><p><b>RESULTS</b>Johnsen score kept stable in the control group at all stages (P > 0.05), but higher than in the model groups at 3 and 4 weeks (P < 0.01). It was lower in the 15 mg/kg than in the 10 mg/kg model group at 4 and 8 weeks (P < 0.01) , and than in the control group at 8 weeks (P < 0.05), but had no significant difference between the 10 mg/kg and the control groups (P > 0.05). Nor did the expression of GATA-4 mRNA in Sertoli cells show any significant difference among the three groups at different stages after the treatment (P > 0.05), and that of GDNF mRNA at different stages in the control group (P > 0.05). Compared with the controls, the level of GDNF mRNA in Sertoli cells was significantly higher at 3 weeks but lower at 4 weeks in the model groups (P < 0.01), and lower in the 15 mg/kg group (P < 0.01) and comparable in the 10 mg/kg group at 8 weeks (P > 0.05); and it was lower in the 15 mg/kg than in the 10 mg/kg group at all stages (P < 0.01).</p><p><b>CONCLUSION</b>Two-dose intraperitoneal injection of 10 mg/kg busulfan at the interval of 24 days is an optimal option for the establishment of a murine model of spermatogenesis regeneration. Higher dose of busulfan may induce deficient expression of GDNF in Sertoli cells and result in incomplete restoration of spermatogenesis.</p>

Clinical analysis of influencing factors of fluid extravasation in patients with calyceal calculi treated by ureteroscopic holmium laser lithotripsy / 中华泌尿外科杂志

Objective To explore the underlying clinical factors and precautionary measures of fluid extravasation in patients with calyceal calculi treated by ureteroscopic holmium laser lithotripsy.Methods A retrospective review was made on clinical records of 138 patients with calyceal calculi receiving retrograde ureteroscopic holmium laser lithotripsy from May 2005 to March 2009. The relevance was studied between the occurance of fluid extravasation complications and various clinical factors using x2 test and binary Logistic regression. The clinical factors included patients' sexes, age groups (＜30 years, 30-50 years, ＞50 years), history of treatment (ESWL or open surgery) for upper urinary tract calculi, preoperative upper urinary tract infection, intraoperative placement of ureteral catheter and the length of procedure duration (＜ 50 min, 50-80 min, ＞ 80 min). Results Fluid extravasation complications occurred in 24 patients. The sexes and age groups were irrelevant to the occurance of fluid extravasation complications; while history of ESWL or open surgery and preoperative infection in upper urinary tract, without intraoperative ureteral catheter placement and long duration of procedure were responsible for the higher rates of the fluid extravasation complications.Conclusion Reasonable selection of patients and timing of operation, regular intraoperative ureteral catheter placement and control the length of procedure duration help to reduce fluid extravasation during retrograde ureteroscopic lithotripsy.

Mouse testicular gene expression pattern differences between spermatogonial stem cell proliferative and differential stages / 中华泌尿外科杂志

Objective To detect the mouse testicular gene expression pattern differences be-tween spermatogonial stem cell (SSC) proliferative and differential stages and study the molecular reg-ulation mechanism in SSC proliferation and differentiation. Methods With the interval of 24 days, male Kunming mice were injected intraperitoneally with two doses of busulfan (10 mg/kg) to establish spermatogenesis regeneration models. 36 k Mouse Genome Array was used to detect the differential gene expression profiles between the stages of SSC proliferation and differentiation. Bioinforrnsties analysis was conducted in GO (gene ontology) and KEGG (Kyoto Encyclopedia of Genes and Ge-nomes) pathway to describe the potential roles that may play in spermatogonial stern cells behavior regulation. Results Nine hundred and eleven differential expression genes were identified by gene arrays in mice testes, consisting of 608 up-regulated and 303 down-regulated in SSC proliferation stage and SSC differentiation stage. The differential expression genes were classified by their biological process, molecular function and cellular component, respectively. Alterations with statistical signifi-cance (P<0.05)appeared in 84 KEG(;signal pathways, including Notch and Wnt signaling pathways which had been proved to be important for stem cell maintenance. Fifty-six differential expression genes were selected as genes related to stem cells, among which 40 genes were up-regulated, including some stem cell biomarkers(such as Cd9, StraS, hgbl-, Oct4 and Thyl)and some growth factors(such as Fgf2, Pdgfa and Csfl). Conclustion The regulation of SSC proliferation and differentiation involves inmany differentially expressed genes in various signal pathways. This study provides a molecular basis for the elucidation of the molecular mechanism behind self-renewal and differentiation of spermatogonial stem cells.

Long-term culture and transplantation of spermatogonial stem cells from BALB/c mice / 中华泌尿外科杂志

Objective To establish a long-term culture system for mouse spermatogonial stem cells(SSCs). Methods Testis cells from 4-6 days postpartum male transgenic BALB/C mice were collected by a modified two-step enzymatic digestion method.After three differential adherence selections,the enriched germ cells were finally suspended in StemPro-34 SFM medium supplemented with other nutrients factors and plated on mouse embryonic fibroblast(MEF)feeder layer.20 ng/ml Glial cell line-derived neurotrophic factor,10 ng/ml basic fibroblast growth factor and 200 ng/ml GDNF-family receptor al were added to the serum-free medium to promote SSCs proliferation.Aduh male BALB/C mice,4-5 weeks old,underwent intraperitoneal injection of 40 mg/kg busulfan as recipient mice.Cultured SSCs were also injected into the seminiferous tubules of the left recipient testis through micromanipulator and right testis as self-control.Testes of recipient mice were observed by a fluorescence stereomicroscope and HE stains at 2 months after transplantation. Results By improved digestion method,the vitality of isolated testis cells was more than 98%and the stem cells was enriched about 18.5 fold. 1-2 days after transferred to MEF feeder, the round germ cells started to proliferate and had the shape of paired or aligned undifferentiated spermatogonia connected by cytoplasmic bridges. After 3-4 days, SSCs proliferated continuously and formed typical colonies. SSCs from BALB/c mice could be cultured and passaged in a steady state for 3 months. Cryostat section through the transplanted testis showed that most of seminiferous tubules were filled with germ cells expressing EGFP.HE staining further showed clearly that seminiferous tubules contained complete spermatogenesis.Conclusions SSCs from BALB/c mice could be cultured in an improved culture system for 3 months.The culture system could facilitate understanding the regulatory mechanism that governs SSCs and might provide an opportunity for the cure of infertility.

Long-term culture and identification of spermatogonial stem cells from BALB/c mice in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To establish a long-term culture system for mouse spermatogonial stem cells (SSCs) and to discuss the key factor that supports mouse SSC self-renewal and proliferation.</p><p><b>METHODS</b>Testis cells from 4-6 days postpartum male transgenic BALB/c mce were collected by a modified two-step enzymatic digestion method and plated on 0. 2% elatin-coated tissue culture plates. The germ cells were enriched by differential adherence selections after respectively incubated for 1, 5 and 24 h and then plated on the mitomycin C-inactivated mouse embryonic fibroblast (MEF) feeder layer. The basal culture medium was StemPro-34 SFM supplemented with other 15 nutrient factors. The 20 ng/ml Glial cell line-derived neurotrophic factor (GDNF), 10 ng/ml basic fibroblast growth factor (bFGF) and 200 ng/ml GDNF-family receptor alpha 1 (GFRalpha1) were added to the serum-free medium to promote SSC proliferation. Several important surface markers and special genes were examined by immunocytochemical staining and RT-PCR analysis.</p><p><b>RESULTS</b>After 3-4 days culture on the MEF feeder, SSCs proliferated continuously and formed typical colonies. SSCs from the BALB/c mice could be cultured in a steady state for 3 months. Immunocytochemical staining showed that Oct4 was specifically expressed in the cultured SSC nucleus and GFRalpha1 strongly expressed on the surface of the membrane. RT-PCR confirmed that the cultured SSCs expressed Oct-4, GFRalpha1, Sox2 and several other special genes resembling undifferentiated spermatogonia.</p><p><b>CONCLUSION</b>SSCs from BALB/c mice could be cultured in the improved culture system for 3 months. This culture system could help further understand the regulating mechanism of SSCs and might provide an opportunity for the treatment of male infertility by SSC transplantation.</p>

Sorting of spermatogonial stem cells in mice / 中华男科学杂志

<p><b>OBJECTIVE</b>To search for an effective method for enriching spermatogonial stem cells in mice.</p><p><b>METHODS</b>Bilateral artificial cryptorchidism was performed on 20 six-week old male Kunming mice. Three months after the operation, the testes were removed and single cell suspension prepared by two-step enzyme digestion. FITC-conjugated anti-alpha6-integrin antibody and PE-conjugated anti-c-kit antibody were added for adequate time on ice. Then the cells with low side scatter light-scattering properties were sorted and positively stained for alpha6-integrin and negative c-kit expression. And the viability of the isolated cells was assessed by trypan blue exclusion.</p><p><b>RESULTS</b>The sorted spermatogonial stem cells constitute 2.8% of the testis cells and over 95% of them were viable.</p><p><b>CONCLUSION</b>FACS can be used to isolate quantities of viable spermatogonial stem cells.</p>

Feasibility of spermatogonial stem cells separation with alpha 6-integrin and c-kit as specific surface makers in mice / 中国组织工程研究

BACKGROUND:It is accepted that the best method for spermatogonial stem cells separation is using artificial cryptorchism model combined with surface makers.OBJECTIVE:To explore the feasibility of separation spermatogonial stem cells with ?6-integrin and c-kit as specific surface markers.DESIGN,TIME AND SETTING:The randomized control experiment was performed at the Renmin Hospital of Wuhan University from May to December 2006.MATERIALS:Forty adult,white Kunming mice with 6 weeks old were randomly divided into cryptorchidism and control groups,with 20 animals in each group.METHODS:Artificial cryptorchidism model was prepared by made an incision at the median of abdomen,and testis was pulled into abdominal cavity,which was fixed at the each side of lateral abdominal wall.There was no treatment in the control group.The single cell suspension of seminiferous epithelium was obtained by traditional two step enzyme digestion at 2-3 months after operation.FITC-conjugated anti-?6-intergrin antibody and PE-conjugated anti-c-kit antibodies were added.Then the cells with low side scatter light-scattering properties were sorted and positively stained for ?6-intergrin and negative c-kit expression.Meanwhile,the viability of the isolated cells was assessed by trypan blue staining.MAIN OUTCOME MEASURES:The morphological changes of cryptorchidism,and the sorting results of spermatogonial stem cells.RESULTS:Cell distribution in seminiferous tubule was disorder with reduced numbers.The layer and lumens were disappeared,and cell division phase could be seen in the center of tubules.Compared to the control group,the testicular cells in the cryptorchidism group were increased in the side scatterlow,Forward scatterhi areas,with figure left-upward displacement.The distribution of ?6-integrin+ and c-kit cells were deviated each other,it named that most ?6-integrin+ cell were not spermatogonial stem cells,so do the c-kit-cells.Only 2.8% of testicular cells exhibited side scatterlow,?6-integrin+,and c-kit-,which were spermatogonial stem cells in the cryptorchidism group.And trypan blue staining showed that over 95% of them were viable.CONCLUSION:Using the two surface markers to sort spermatogonial stem cells can advance the purity of the spermatogonial stem cells in cell suspension,but the specificity is insufficient.

Treatment of buried penis in children with fixation at the dartos coat of penis through prepubic route / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the long-time results of the treatment of buried penis with fixation at the base of penis through pre-pubic route.</p><p><b>METHODS</b>From Aug 2002 to Dec 2003, the procedure was performed in 34 children without penile skin insufficiency. The major technique involved the release of hided penis shaft by partly shearing suspensory ligament, fixating the bucks fascia and tunica albuginea at 3 and 9 o'clock positions with the subcutaneous penile skin at the base of penis just under the pubic symphysis.</p><p><b>RESULTS</b>Twenty-one patients have been reviewed over one year with satisfactory appearance. Thirteen patients were lost to follow-up. No recurrence happened in our group.</p><p><b>CONCLUSION</b>We compare this technique with major treatment methods for buried penis in children at present, and we conclude this technique is a reasonable, simplified and aesthetical method for the treatment of buried penis in children.</p>