Triptorelin relieves lower urinary tract symptoms in Chinese advanced prostate cancer patients: a multicenter, non-interventional, prospective study.

BACKGROUND: Although triptorelin is increasingly used in China for biochemical castration, its effects on primary prostate cancer symptoms remain unclear. This study aimed to assess the prevalence of lower urinary tract symptoms (LUTS) in Chinese prostate cancer patients and the effectiveness of triptorelin on LUTS. METHODS: In this 48-week multicenter, non-interventional, prospective study, we enrolled patients with locally advanced or metastatic prostate cancer. Patients received triptorelin (15 mg) intramuscularly at baseline and at weeks 12, 24, and 36 with symptom assessment using the International Prostate Symptoms Score (IPSS). The primary endpoints were the prevalence of LUTS at baseline per IPSS categories and the percentage of patients with moderate to severe LUTS (IPSS > 7) at baseline, having at least a 3-point reduction of IPSS score at week 48. RESULTS: A total of 398 patients were included; 211 (53.0%) and 160 (40.2%) among them had severe and moderate LUTS, respectively. Of the patients with IPSS scores available at baseline and at week 48 (n = 213), 81.2% achieved a reduction in IPSS of at least 3 points. Of the patients with moderate to severe LUTS at baseline and IPSS scores available at baseline and at week 48 (n = 194), 86.6% achieved a total IPSS reduction of at least 3 points. CONCLUSIONS: The vast majority of Chinese patients with locally advanced or metastatic prostate cancer scheduled to receive triptorelin as part of their standard treatment have severe or moderate LUTS. Triptorelin therapy resulted in sustained improvement of LUTS in these patients.

[Gujing Maisiha Tablets combined with natural vitamin E for the treatment of idiopathic asthenospermia].

OBJECTIVE: To investigate the therapeutic efficacy and safety of Gujing Maisiha Tablets combined with natural vitamin E in the treatment of idiopathic asthenospermia. METHODS: This study included 135 outpatients with idiopathic asthenospermia received in our hospital from February 2015 to January 2016. We randomly divided them into a treatment group (n = 65, aged 22－44 ï¼»mean 32.8ï¼½ yr) and a control group (n = 55, aged 21－43 ï¼»mean 33.7ï¼½ yr) to be treated with Gujing Maisiha Tablets combined with natural vitamin E or natural vitamin E only, both for 90 days. We obtained total sperm motility and the percentage of progressively motile sperm (PMS) from the patients before and after medication and evaluated the clinical effects by comparing the collected parameters and pregnancy rates between the two groups. RESULTS: The baseline total sperm motility and PMS were (25.23 ± 5.57)% and (17.53 ± 5.78)% in the treatment group, with no statistically significant differences from (26.05 ± 6.77)% and (15.11 ± 6.55)% in the control (P >0.05). After 90 days of medication, both the treatment and the control groups showed remarkable increases in total sperm motility (ï¼»48.73 ± 8.66ï¼½% and ï¼»36.54 ± 8.09ï¼½%, P <0.05) and PMS (ï¼»32.77 ± 6.04ï¼½% and ï¼»26.99 ± 6.87ï¼½%, P <0.05). However, both total sperm motility and PMS were significantly higher in the treatment than in the control group after medication (P <0.05), and so was the total rate of clinical effectiveness (73.85% vs 54.55%, P <0.05). No adverse reactions were observed in either of the two groups during the treatment. CONCLUSIONS: Gujing Maisiha Tablets combined with natural vitamin E is safe and effective for the treatment of idiopathic asthenospermia.

Transurethral endoscopic treatment of seminal vesicle cysts (report of seven cases).

OBJECTIVE: Seminal vesicle cysts are a rare disorder of the male reproductive system. The goal of this report was to summarize the radiological manifestations and transurethral endoscopic treatment of large seminal vesicle cysts. MATERIALS AND METHODS: The clinical data of seven cases of giant seminal vesicle cysts, including their symptoms, radiological images, transurethral endoscopic treatment, and postoperative follow-up, were retrospectively reviewed. RESULTS: Computerized tomography and magnetic resonance imaging (MRI) demonstrated the cysts behind the bladder, above the prostate, and away from the midline. The lesions ranged in size from 8.26 cm × 7.98 cm × 4.85 cm to 9.27 cm × 8.95 cm × 8.15 cm. Two cases were associated with ipsilateral renal and ureteral agenesis and were classified as congenital malformations. The other five cases were simple seminal vesicle cysts thought to be secondary to acquired ejaculatory duct obstruction. All seven cases were successfully treated using transurethral endoscopic unroofing with cautery of the mucosa. All the seminal vesicle cysts were confirmed by pathologic examination. No malignant disease was found. All preoperative symptoms resolved after surgery. No complications were observed. No patient developed abnormalities of erection, ejaculation, or orgasm. No bladder or rectal injuries were noted. The seminal vesicle cysts were significantly decreased in size or absent 3-6 months after treatment. CONCLUSION: MRI best characterized seminal vesical cysts and their cause. Transurethral unroofing with cautery of the mucosa is an extension of well-accepted cystoscopic techniques. It is safe, easy to perform, and effective. It is the preferred method for the treatment of large seminal vesicle cysts.

Reduction of fertility in male mice immunised with pSG.SS.C3d3.YL.Bin1b recombinant vaccine.

OBJECTIVES: To study the anti-fertility effect of a DNA vaccine using Bin1b as the target antigen in male mice. METHODS: A novel recombinant eukaryotic vector containing a fusion gene sequence of mouse Bin1b in tandem with three copies of C3d fragment (C3d3) was used to construct pSG.SS.C3d3.YL.Bin1b. The correct expression of the Bin1b-C3d3 protein was confirmed in transfected HEK293 cells by indirect immunofluorescence and western blot analysis. The fertility of immunised mice was determined by a mating experiment and sperm motility test. Anti-Bin1b antibody titres in sera were examined by ELISA assays. Binding activity of C3d3 fragment of the fusion protein was verified in C3d receptor-expressing Raji cells and flow cytometric analysis. RESULTS: Immunisation of pSG.SS.C3d3.YL.Bin1b recombinant DNA vaccine significantly decreased sperm motility and compromised fertility in male mice. ELISA results showed that the titres of anti-Bin1b IgG in sera of immunised mice increased markedly with the immunisation process. Further, the anti-fertility effect of pSG.SS.C3d3.YL.Bin1b was significantly better than that of pSG.SS.YL.Bin1b DNA vaccine and generated higher titres of anti-Bin1b antibody. CONCLUSIONS: Our results show that recombinant DNA vaccine targeting Bin1b can markedly reduce fertility in male mice, providing an alternative approach for birth control.

Safety and efficacy of levofloxacin versus ciprofloxacin for the treatment of chronic bacterial prostatitis in Chinese patients.

Levofloxacin is a synthetic fluoroquinolone that is usually used to treat chronic bacterial prostatitis. We investigated the safety and efficacy of levofloxacin compared with ciprofloxacin for the treatment of chronic bacterial prostatitis in Chinese patients. This was a multicenter, open-label, randomized controlled non-inferiority trial. Four hundred and seventy-one patients with clinical symptoms/signs were enrolled into the study, and 408 patients were microbiologically confirmed chronic bacterial prostatitis, who were randomized to either oral levofloxacin (500 mg q.d.) or ciprofloxacin (500 mg b.i.d.) for 4 weeks. Bacterial clearance rate, clinical symptoms/signs, adverse reactions and disease recurrence were assessed. The clinical symptoms and signs (including dysuria, perineal discomfort or pain) and bacteria cultures in 209 patients treated with levofloxacin and 199 patients treated with ciprofloxacin were similar. The most common bacteria were Escherichia coli and Staphylococcus aureus. One to four weeks after the end of 4 weeks treatment, the bacterial clearance rate (86.06% vs. 60.03%; P<0.05) and the clinical efficacy (including clinical cure and clinical improvement(93.30% vs. 71.86%; P<0.05)) were significantly higher in the levofloxacin-treated group than in the ciprofloxacin-treated group. The microbiological recurrence rate was significantly lower in the levofloxacin-treated group than in the ciprofloxacin-treated group (4.00% vs. 19.25%; P<0.05). Rates of adverse events and treatment-related adverse events were slightly lower in the levofloxacin-treated group than in ciprofloxacin-treated group. Levofloxacin showed some advantages over ciprofloxacin in terms of clinical efficacy and disease recurrence, with a low rate of adverse events, for the treatment of chronic bacterial prostatitis in Chinese patients.

Resveratrol induces apoptosis associated with mitochondrial dysfunction in bladder carcinoma cells.

OBJECTIVE: Resveratrol shows chemopreventive activity in a variety of human cancers by targeting mitochondria and triggering apoptosis. The purpose of this study was to investigate the antitumor action of resveratrol in bladder cancer and its underlying mechanism. METHODS: Using two different bladder cell lines, BTT739 and T24, the cytotoxicity of resveratrol were determined by MTT assay. The apoptosis induced by resveratrol was assayed by transferase dUTP nick end labeling staining. To show whether the mitochondrial dysfunction involved in the effects of resveratrol, mitochondrial function was detected by mitochondrial membrane potential, reactive oxygen species production and adenosine 5'-triphosphate content. In addition, the markers of apoptosis in the intrinsic mitochondrial-dependent pathway were analyzed by the release of cytochrome c and the activities of caspase-9 and caspase-3. RESULTS: Resveratrol effectively decreased cell viability and induced apoptosis in a concentration- and time-dependent manner. In addition, resveratrol significantly disrupted the mitochondrial membrane potential in both intact cells and isolated mitochondria. Resveratrol also increased reactive oxygen species production and reduced adenosine 5'-triphosphate concentrations. Western blot analysis showed that resveratrol provoked the release of cytochrome c from mitochondria to the cytosol. Furthermore, resveratrol significantly promoted the activation of caspase-9 and caspase-3. CONCLUSIONS: These findings suggest that resveratrol efficiently triggers apoptosis in bladder cancer cells through the intrinsic mitochondrial-dependent pathway, which is associated with mitochondrial dysfunction. Resveratrol might have great pharmacological promise in the treatment of bladder cancer.

A novel DNA/peptide combined vaccine induces PSCA-specific cytotoxic T-lymphocyte responses and suppresses tumor growth in experimental prostate cancer.

OBJECTIVE: To develop a completely novel DNA peptide-combined vaccine and determine whether it can efficiently improve tumor-specific cytotoxic T lymphocyte (CTL) responses and inhibit tumor progression in experimental prostate cancer models. METHODS: The DNA/peptide combined vaccine was prepared by the self-assembly of a cationic peptide ([K]18P9) containing 18 lysines and a CTL epitope peptide, prostate stem cell antigen (PSCA (14-22)) (HLA-A2 restricted) with a recombinant plasmid encoding human full-length PSCA gene (pcDNA3.1(+)-PSCA) through electrostatic interactions. The formation of a DNA/peptide complex was examined by DNA retardation assay, DNase I protection assay, and transmission electron microscopy. The efficacy of vaccination using this complex was demonstrated in terms of the PSCA-specific CTL activity and antitumor immunity to PSCA(+) tumors in a murine model. RESULTS: This form of DNA/peptide complex could efficiently transfer the plasmid encoding full-length PSCA gene into mammalian cells and induced potent CTLs cytotoxicity against a human prostate carcinoma cell line established from the left supraclavicular lymph node metastasis from a 50-year-old man with prostate carcinoma in 1977. Expressing PSCA compared with pcDNA3.1(+)-PSCA, [K]18P9 peptide, or pcDNA3.1(+). Moreover, the vaccination of mice with this complex induced a potent antitumor immunity to prostate carcinomas in a xenograft tumor model in nude mice. CONCLUSION: This study suggests that a specific antitumor immune response can be induced by this DNA/peptide combined vaccine, which represents a new strategy for use in the immunotherapy of prostate cancer.

Imaging diagnosis, transurethral endoscopic observation, and management of 43 cases of persistent and refractory hematospermia.

The goal of this study was to explore minimally invasive transurethral imaging and surgery for the treatment of severe, persistent hematospermia in cases that were refractory to conservative treatments. The study included 43 patients (aged 22-77 years; average, 44.6 years) with long-lasting, severe hematospermia, accompanied by discomfort or pain in the lumbosacral or perineal region, dysuria, frequent micturition, decreased semen volume, and/or azoospermia. Patient symptoms had persisted for 1 to 10 years (average, 5.3 years). Computed tomography or magnetic resonance imaging of each patient was evaluated, and transurethral surgery was performed. The causes of hematospermia were identified in all 43 patients, and their ejaculatory duct obstruction or seminal vesiculitis was successfully treated. No serious intraoperative or postoperative complications occurred. Pathologic analyses revealed that all of the resected or biopsied seminal vesicle tissues had chronic nonspecific inflammation in the seminal vesicle wall, and no tumors were identified. Preoperative symptomology of hematospermia disappeared in all patients followed up for 2 to 30 months (average, 16 months). A single patient experienced recurrence at 11 months and had a second minimally invasive surgery that was curative. A total of 95.3% (41 of 43) of the patients experienced normal orgasmic intensity after surgery. Magnetic resonance imaging is a valuable and accurate diagnostic method for the identification of causative factors underlying hematospermia. Transurethral dilation of ejaculatory ducts, incision of the verumontanum or the distal end of the ejaculatory ducts, and incision or resection of the relevant cysts represent simple, safe, and reliable approaches for the management of refractory cases of hematospermia that do not respond to conservative treatments.

Transumbilical laparoendoscopic single-site renal pedicle lymphatic disconnection for refractory chyluria.

PURPOSE: To report our preliminary techniques and experience with transumbilical laparoendoscopic single-site renal pedicle lymphatic disconnection (LESS-RPLD) in seven patients with refractory chyluria. PATIENTS AND METHODS: Between June 2009 and September 2010, seven patients with refractory chyluria underwent LESS-RPLD. In the patients, a 2- to 3-cm single inverted U-shaped supraumbilical incision was made, and a homemade single multichannel port using a surgical glove and three conventional trocars was placed into the abdominal cavity. Flexible electric coagulation hook and pliers were used for renal pedicle dissection. A straight ultrasound knife was used for lymphatic disconnection. RESULTS: All the operations were successfully completed without conversion to open surgery, although an additional 3-mm trocar was used to push the liver in one patient. The mean operative time was 125 (96-165) minutes. The mean blood loss was estimated to be 112 (50-250) mL. Chyluria disappeared in all patients after surgery and did not recur during the follow-up period (3-15, mean 8.3 mos). CONCLUSION: LESS-RPLD is safe and feasible, with favorable short-term outcomes and aesthetic effect.

FSCB phosphorylation in mouse spermatozoa capacitation.

It is generally accepted that spermatozoa capacitation is associated with protein kinase A-mediated tyrosine phosphorylation. In our previous study, we identified the fibrous sheath CABYR binding protein (FSCB), which was phosphorylated by PKA. However, the phosphorylation status of FSCB protein during spermatozoa capacitation should be further investigated. To this aim, in this study, we found that phosphorylation of this 270-kDa protein occurred as early as 1 min after mouse spermatozoa capacitation, which increased over time and remained stable after 60 min. Immunoprecipitation assays demonstrated that the tyrosine and Ser/Thr phosphorylation of FSCB occurred during spermatozoa capacitation. The extent of phosphorylation and was closely associated with the PKA activity and spermatozoa motility characteristics. FSCB phosphorylation could be induced by PKA agonist DB-cAMP, but was blocked by PKA antagonist H-89.Therefore, FSCB contributes to spermatozoa capacitation in a tyrosine-phosphorylated format, which may help in further elucidating the molecular mechanism of spermatozoa capacitation.

[One-hole high ligation of internal spermatic vein by embryonic natural orifice transumbilical laparoscopy: a recommendable treatment of varicocele].

OBJECTIVE: To evaluate the one-hole method for high ligation of the internal spermatic vein by embryonic natural orifice transumbilical laparoscopy. METHODS: We used the one-hole method for high ligation of the internal spermatic vein by embryonic natural orifice transumbilical laparoscopy in the treatment of 15 cases of varicocele, 13 in the left and 2 in the right side, and appraised the treatment results by follow-up 1 month after the surgery. RESULTS: All the operations succeeded and no complications developed. The average operation time was 28 minutes and the mean hospital stay was 4 days. Symptoms were significantly relieved in all the patients and the scars were inconspicuous at follow-up. CONCLUSION: The one-hole method is a novel option for high ligation of the internal spermatic vein by embryonic natural orifice transumbilical laparoscopy in the treatment of varicocele. It is recommendable for its advantages of simple procedure, less pain, few complications, quick recovery, mini-invasiveness and cosmetic acceptability.

Construction of a DNA vaccine encoding Flk-1 extracellular domain and C3d fusion gene and investigation of its suppressing effect on tumor growth.

Although the critical role of complement component C3d as a molecular adjuvant in preventing virus infection is well established, its role in cancer prophylaxis and treatment is unclear. In this study, we constructed a recombinant plasmid encoding Flk-1 and C3d3 fusion proteins and investigated its transient expression in vitro in transfected eukaryotic cells and its antibody response in immunized mice. Subsequently, we investigated the vaccine's ability to elicit an immune response leading to suppression of angiogenesis and tumor growth in mice bearing bladder transitional cell carcinoma. Using Western blotting, immunocytochemistry, and flow cytometry, we detected the expression of Flk-1 and C3d3 fusion proteins in COS-7 cells transfected with these recombinant plasmids. Further binding experiment using CR2 (C3d receptor) positive Raji cells that were incubated with transfected COS-7 supernatant indicated that C3d was successfully fused to Flk-1. Although both vaccines elicited peak antibody levels at 5 weeks, Flk-1-specific antibody titer in pSG.SS.Flk-1(ECD).C3d3.YL-immunized mice was significantly higher when compared to pSG.SS.Flk-1(ECD).YL-immunized mice. The results of experiments with bladder tumor-bearing mice showed that the vaccine inhibited tumor growth significantly. These results suggest that C3d plays a critical role in tumor immunotherapy by promoting antibody response in Flk-1-based DNA vaccines. This approach may provide a new strategy for the rational design of anti-angiogenic therapies for the treatment of solid tumors and provide a basis for the further exploitation and application of the anti-angiogenesis DNA vaccines.

[Urological complications in 1223 kidney transplants].

OBJECTIVE: To review retrospectively the urological complications in 1 223 kidney transplants. METHODS: A total of 1 223 kidney transplants were divided into ureteroneocystostomy group (n = 948) and ureteroureterostomy group (n = 275) according to the methods of urinary tract reconstruction. The incidence and management of urological complications such as urinary fistula, obstruction of ureter, vesicoureteral reflux (VUR) and urinary tract infection (UTI) were summarized respectively. RESULTS: Overall, urological complications were encountered in 217 (17.7%) cases, including 43 cases of urinary fistula (3.5%), 35 obstruction of ureter (2.9%), 14 VUR (1.1%) and 125 UTI (10.2%). Urinary fistula was 39 (4.1%) cases and 4 cases (1.5%) (P < 0.05), obstruction of ureter 22 (2.3%) & 13 (4.7%) (P < 0.05), VUR 14 (1.5%) & 0 (0%) (P < 0.05) and UTI 109 (11.5%) & 16 (5.8%) (P < 0.01) in the ureteroneocystostomy group and ureteroureterostomy group respectively. Seventy patients underwent surgical treatment. The 3-year survival rate of graft with urological complications and without urological complications were 82.3% and 84.7% respectively. CONCLUSIONS: Ureteroureterostomy can decrease the incidence of urological complications after kidney transplantation. Most of urological complications require surgical interventions. The long-term graft survival is not affected by a correctly treated urological complication.

Relationship between glucocorticoid receptor signal pathway and androgen-independent prostate cancer.

OBJECTIVE: To study the role of the glucocorticoid receptor (GR) signal pathway and downstream cytokine interleukin-6 (IL-6) in androgen-independent growth of prostate cancer (PC). METHODS: The human androgen-dependent PC (ADPC) cell line LNCaP and androgen-independent PC (AIPC) cell line DU145 were cultured in vitro. Immunocytochemistry was used to examine the expression of the androgen receptor (AR), GR, HSP90 and IL-6. The GR antagonist RU486 was used to treat cultured cells, and the effects of RU486 on the proliferation of both cell lines were analyzed by MTT assay. Expression of HSP90 and IL-6 mRNA and protein was assessed by RT-PCR and Western blots, respectively. RESULTS: LNCaP cells were AR-positive and GR-negative, whereas DU145 cells were GR-positive and AR-negative. The expression of HSP90 and IL-6 in DU145 cells were significantly stronger than that in LNCaP cells (p < 0.01). RU486 had no obvious effects on the growth of LNCaP cells, but exerted a significant time- and dose-dependent growth inhibition on DU145 cells at doses as low as 0.1 micromol/l. RU486 treatment of DU145 cells also resulted in a dose-dependent decrease in the expression of HSP90 and IL-6 mRNA and protein. CONCLUSIONS: The GR signal pathway may be the main survival pathway for DU145 cells. Abnormal hyperactivation of the GR signal pathway and its promoting the expression of HSP90 and IL-6 contribute to the progression of ADPC to AIPC after androgen ablation.

[Testis homotransplantation: a report of 12 cases].

OBJECTIVE: To evaluate the effect of testis homotransplantation in the treatment of androgen deficiency and infertility. METHODS: We retrospectively analyzed 12 cases of testis homotransplantation. RESULTS: Surgical success was achieved in 11 cases, all with a significantly increased level of serum testosterone, and markedly improved secondary sex characteristics and sexual function. CONCLUSION: Testis homotransplantation is highly effective for the treatment of androgen deficiency in males, but has little effect on spermatogenesis.

[Construction and expression of eukaryotic recombinant of mouse fertilin beta subunit in HEK293 cells].

OBJECTIVE: To construct the eukaryotic expression recombinant pSG. SS. C3d3. YL-Fbeta and analyze the expression of mouse fertilin beta subunit in HEK293 cells. METHODS: The cDNA fragment expressing the disintegrin domain of mouse fertilin beta was obtained by PCR, and then inserted into the eukaryotic plasmid pSG. SS. C3d3. YL to get recombinant plasmid pSG. SS. C3d3. YL-Fbeta, which was transfected into the HEK293 cell line to express the target protein Fbeta after identified by restriction enzyme digestion. And then Fbeta was detected by indirect immunofluorescence through confocal laser scanning microscopy, Western blot, immunohistochemistry staining and flow cytometry assay. RESULTS: The recombinant vector pSG.SS.C3d3. YL-Fbeta could express Fbeta in HEK293 cells. CONCLUSION: The expression of Fbeta in eukaryotic cells provides a foundation for further researches on the effect of high F, expression on fertilization process.

[High throughput screening and analysis of prostate cancer-related genes through mining databases].

BACKGROUND & OBJECTIVE: Investigation of differentially expressed genes in prostate cancer tissues may help to understand the molecular mechanism of prostate cancer and provide diagnostic markers or new targets for therapy. The Cancer Genome Anatomy Project (CGAP) public database provides an unprecedented opportunity for cancer researchers to mine genes differentially expressed in cancer tissues by bioinformatic methods. This study was to explore the feasibility of incorporating the Internet-available Serial Analysis of Gene Expression (SAGE) and cDNA databases to find human prostate cancer-related genes. METHODS: SAGE digital gene expression displayer (DGED) and cDNA DGED were used to analyze differentially expressed (>5 folds) genes in malignant prostate tissues compared with normal prostate tissues. The SAGE tags were filtered by their confidence and our 3 criteria. To test the confidence of tags of virtual digital analysis for gene identification, the modified GLGI (generation of longer cDNA fragments from SAGE tags for gene identification) was used to get the cDNA 3' end downstream of 20 tags. Main functions of all candidate genes and their relations to prostate cancer were annotated. RESULTS: Fifty-three differentially expressed genes were screened out by SAGE DGED, 26 of them were up-regulated and 27 were down-regulated in prostate cancer; 28 differentially expressed genes were got by cDNA DGED, 15 of them were up-regulated and 13 were down-regulated in prostate cancer. CONCLUSION: Reasonable use of public databases by the Internet-available tools is a simple, effective approach to get cancer-related genes, and might provide useful clues for further investigation although the results require experimental validation.

[Preliminary screening of specific surface markers of human spermatogonial stem cells].

OBJECTIVE: To explore the specific surface markers for the isolation and purification of human spermatogonial stem cells (SSC). METHODS: Specific markers of human SSC were screened and identified in fetal and adult testes by immunohistochemical assay, using HSC markers c-kit, Thy-1 and human ES integrins. RESULTS: In human adult testes, the alpha6 integrin extensively and significantly expressed on the surface of most of the germ cells in the seminiferous tubule, and beta1 integrin mainly expressed on the surface of the germ cells residing on or near the basal membrane in the seminiferous tubule. Thy-1 scattering expressed on the surface of some cells of the basal membrane, and on some Leydig cells as well. The three antigen markers expressed on the SSC of human adult testes specifically to some extent. SSEA-1 specifically expressed on the surface of the gonocytes in the fetal testes. CONCLUSION: The alpha6 and beta1 integrins and Thy-1 may be used for the SSC isolation as positive markers. SSEA-1 can be used as an identification marker for the fetus SSC.

[Long-term survival of human spermatogonial stem cells in vitro and its functional identification].

OBJECTIVE: The culture of human spermatogonial stem cells (SSC) has not been studied in detail yet. Here we tried to explore the optimized culture method of human SSC by using several different co-culture systems. METHODS: The alpha6 +Thy-1 +c-kit- cells acquired by the immunomagnetic beads sorting technique were cultured in different co-culture systems. Their morphological, biological characteristics and survival rates were intensively observed by microscopic or immunocytochemical assay. The long-term survival rate of human SSC during culture period was evaluated by germ cell transplantation technique. RESULTS: The alpha6 +Thy-1 +c-kit- cells could stably survive in the DMEM and DMEM/F12 mediums with fetal bovine serum (FBS) on feeder layer. The survival rates within 1 week were more than 90%. The long-time culture showed the cells were gradually attached on the surface of Sertoli cells by the manner of scattered single cell or accumulated masses. Part of the SSC became more tightly attachment with Sertoli cells or mounted among the Sertoli cells. They could survive or even proliferate for more than 3 months in vitro. Germ cells transplantation study showed that some alpha6 +Thy-1 +c-kit- cells labeled by PKH26 could resided on the basal membrane of seminiferous tubule of nude mice, appearing as single or coupled cells 2 months later after transplantation. The function evaluation of the cultured cells by counting the fluorescent cells in the seminiferous tubule showed 54.9% and 9.2% of SSC in the alpha6 +Thy-1 +c-kit- cells were still remained after cultured for 2 and 4 weeks, respectively. CONCLUSION: Human SSC could maintain survival in vitro for more than 3 months, but it was still need to seek for a more optimized and successful culture system for its efficient expansion and proliferation. Thus it will open up a wide prospect for the understanding of the biology of human SSC and the treatment of male sterility.