The value of urinary exosomal microRNA-21 in the early diagnosis and prognosis of bladder cancer.

Bladder cancer (BC) poses high morbidity and mortality, with urinary exosomal microRNA (miR)-21 showing potential value in its diagnosis and prognosis, and we probed its specific role. We prospectively selected 116 BC patients and 116 healthy volunteers as the BC and control groups, respectively. BC urinary exosomal miR-146a-5p, miR-93-5p, miR-663b, miR-21, and miR-4454 relative expression levels were assessed. The correlations between clinical indexes and urinary exosomal miR-21, prognostic value of miR-21, and diagnostic value of the five candidate miRNAs, urine cytology, and miRNA joint diagnostic panel for BC and urinary exosomal miR-21, miR-4454, and urine cytology for Ta-T1 and T2-T4 stage BC were analyzed. Urinary exosomal miR-146a-5p, miR-93-5p, miR-663b, miR-21, and miR-4454 were highly expressed in BC patients. miR-146a-5p, miR-93-5p, miR-663b, miR-21, miR-4454, miRNA combined diagnostic panel, and urine cytology had certain diagnostic value for BC, with miR-21, miR-4454, and miRNA co-diagnostic panel showing the highest diagnostic value. Collectively, urinary exosomal miR-21 was closely related to Tumor-Node-Metastasis staging and grading in BC patients. Urinary exosomal miR-21 had high diagnostic value for BC and Ta-T1 and T2-T4 stage BC, and had high predictive value for BC poor prognosis, providing an effective indicator for the occurrence, development, and prognostic assessment of BC.

Dentin matrix protein 1 and phosphate homeostasis are critical for postnatal pulp, dentin and enamel formation.

Deletion or mutation of dentin matrix protein 1 (DMP1) leads to hypophosphatemic rickets and defects within the dentin. However, it is largely unknown if this pathological change is a direct role of DMP1 or an indirect role of phosphate (Pi) or both. It has also been previously shown that Klotho-deficient mice, which displayed a high Pi level due to a failure of Pi excretion, causes mild defects in the dentinal structure. This study was to address the distinct roles of DMP1 and Pi homeostasis in cell differentiation, apoptosis and mineralization of dentin and enamel. Our working hypothesis was that a stable Pi homeostasis is critical for postnatal tooth formation, and that DMP1 has an antiapoptotic role in both amelogenesis and dentinogenesis. To test this hypothesis, Dmp1-null (Dmp1(-/-)), Klotho-deficient (kl/kl), Dmp1/Klotho-double-deficient (Dmp1(-/-)/kl/kl) and wild-type (WT) mice were killed at the age of 6 weeks. Combinations of X-ray, microcomputed tomography (µCT), scanning electron microscopy (SEM), histology, apoptosis and immunohistochemical methods were used for characterization of dentin, enamel and pulp structures in these mutant mice. Our results showed that Dmp1(-/-) (a low Pi level) or kl/kl (a high Pi level) mice displayed mild dentin defects such as thin dentin and a reduction of dentin tubules. Neither deficient mouse line exhibited any apparent changes in enamel or pulp structure. However, the double-deficient mice (a high Pi level) displayed severe defects in dentin and enamel structures, including loss of dentinal tubules and enamel prisms, as well as unexpected ectopic ossification within the pulp root canal. TUNEL assay showed a sharp increase in apoptotic cells in ameloblasts and odontoblasts. Based on the above findings, we conclude that DMP1 has a protective role for odontoblasts and ameloblasts in a pro-apoptotic environment (a high Pi level).

[Protective effect of baicalin on experimental periodontitis in rats and its possible mechanisms].

OBJECTIVE: To investigate the effect of baicalin on the experimental periodontitis in rats, as well as the expression of MMP-1, MMP-2, MMP-9. METHODS: Twenty-seven adult male Sprague-Dawley rats were divided into three groups, with 9 rats in each group. A nylon thread was placed around the lower first molars of rats, which were sacrificed after 7 days. Baicalin (200 mg/kg) was administered to the experimental group by oral gavage, starting one day before the induction of periodontitis. The negative control group received vehicle (0.5% carboxymethylcellulose) alone. The blank control group did not get induction of periodontitis. The alveolar bone loss (ABL) and the area fraction (AA% ) occupied by collagen fibers were assessed. MMP-1, MMP-2 and MMP-9 protein expressions in the gingiva were detected by immunohistochemistry. RESULTS: Baicalin treatment significantly decreased ABL compared with the negative control group (P = 0.009). AA% of collagen fibers was significantly higher in baicalin-treated group than in the negative control group (P = 0.047). Baicalin treatment significantly down-regulated the protein expression for MMP-1 (P = 0.023) and MMP-9 (P = 0.042) and decreased the expression for MMP-2 (P = 0.099) compared with the negative control group. CONCLUSIONS: Baicalin protects against tissue damage in ligature-induced periodontitis in rats, which might be mediated in part by its inhibitory effect on the expression of MMP-1, MMP-2 and MMP-9.

[Holmium laser urethrotomy for male urethral stricture].

OBJECTIVE: To evaluate transurethral holmium laser incision, its safety and effect in the treatment of male urethral stricture. METHODS: Thirty-eight males with urethral stricture were treated by 1045 W holmium laser urethrotomy, 18 with the stricture length shorter than 1.0 cm, 9 between 1.0 cm and 1.5 cm, 7 longer than 1.5 cm , 4 with occlusive stricture and 6 companies with bladder calculus. The average peak urinary flow rate (Q(max)) was (5.6 +/- 2.3) ml/s. RESULTS: Successful surgery was achieved in 36 of the cases, with no complications and the average Q(max) increased to (17.5 +/- 3.4) ml/s. Two cases were converted to open surgery. Thirty-two cases were followed up for 3-18 months, of whom 4 received urethral dilation and 2 underwent a second holmium laser urethrotomy. CONCLUSION: Holmium laser urethrotomy is a safe, effective and minimally invasive therapeutic modality for male urethral stricture.

The effect of a single nucleotide polymorphism in the matrix metalloproteinase-1 (MMP-1) promoter on force-induced MMP-1 expression in human periodontal ligament cells.

Matrix metalloproteinase-1 (MMP-1) 1G/2G (-1,607) polymorphisms have been identified and shown to influence the transcription of the MMP-1 gene. In order to compare the expression of MMP-1 with different MMP-1 gene promoter alleles after force loading, human periodontal ligament (PDL) cells were cultured and genotyped into three alleles by polymerase chain reaction and restriction endonuclease cleavage. The three genotypes of PDL cells were centrifuged and the expression of MMP-1 mRNA and protein were determined by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. The results showed that centrifugal force upregulated the expression of both MMP-1 mRNA and protein in all three genotypes of PDL cells. The induction of MMP-1 by force was significantly greater in cells with a 2G/2G genotype or a 1G/2G genotype than in cells homozygous for the 1G allele. The MMP-1 mRNA and protein levels were significantly higher for cells with the 2G allele than for cells with the 1G/2G allele or the 1G allele. These results suggest that a single nucleotide polymorphism in the -1,607 bp MMP-1 promoter region might be associated with the difference observed in the endogenous expression of MMP-1 in PDL cells under mechanical force.

[Small interfering RNA inhibits the expression of connexin43 in the human corpus cavernosum penis smooth muscle cells].

OBJECTIVE: To inhibit the expression of connexin43 (Cx43) in the human corpus cavernosum penis smooth muscle cells by small interfering RNA (siRNA) and detect the gap junction intercellular communication (GJIC), and to investigate the application of siRNA technology in the gap junction of corpus cavernosum penis smooth muscle cells and its role in the penile erection process. METHODS: With the help of the software of Ambion Corporation, the specific recombinant plasmids with siRNA targeting human Cx43 gene were constructed. The recombinant plasmids having been stably transferred into human corpus cavernosum penis smooth muscle cells for 48 hours, semi-quantitive reverse transcription polymerase chain reaction (RT-PCR) and Western blotting techniques were used to examine the inhibitory effects of siRNA on the expressions of the Cx43 gene and protein, in comparison with the siRNA negative control and the blank control group, respectively. The GJIC was detected by scrape-loading and fluorescence dye transfer experiments through the fluorescence microscope. RESULTS: The results of enzyme digestion analysis and DNA sequencing showed that the recombinant plasmid pSilencer 1.0-U6-siRNA-Cx43 was successfully constructed. The relative levels of Cx43 mRNA and protein expression in the smooth muscle cells were (0.45 +/- 0.08)% and (0.56 +/- 0.06)% after successful transfer of the recombinant plasmid. However, the expression levels of mRNA and protein were (0.72 +/- 0.04)% and (0.80 +/- 0.08)% in the negative siRNA transfer group, and (0.74 +/- 0.09)% and (0.77 +/- 0.11)% in the blank control, respectively, with a significant difference (P < 0.05). The GJIC also decreased significantly. CONCLUSION: siRNA can significantly inhibit the expression of Cx43 and block the GJIC in the human corpus cavernosum penis smooth muscle cells. siRNA technology plays an important role in penile erection and flaccidity.

Rose bengal staining in detection of oral precancerous and malignant lesions with colorimetric evaluation: a pilot study.

Early detection of oral precancerous and malignant lesions is still a diagnostic challenge for most of clinicians, and ideal adjuncts for detection of these lesions are currently unavailable. Our preliminary study has indicated that rose bengal (RB) staining might have the potency as a diagnostic aid; however, its clinical significance and reliability in hospital-based population are still not clear. In the present study, we investigated the efficacy of RB staining in detection of oral precancerous and malignant lesions. RB staining was conducted in 132 patients, and staining results were determined by a 4-graded shade guide, which had been quantitatively measured in the 1976 CIEL*a*b* space by instrumental colorimetry. Histological examination was performed in 128 of 132 patients after RB staining. The sensitivity and specificity to detect epithelial dysplasia (DP) and oral squamous cell carcinoma were 93.9 and 73.7%, respectively. The positive and negative likelihood ratios were 3.570 and 0.082, respectively. Moreover, RB staining seemed promising to detect DP in oral leukoplakia, lichen planus and leukokeratosis. In this study, 5 of 6 DP or oral squamous cell carcinoma were identified by RB staining before histological examination. In conclusion, RB staining may be a valuable diagnostic test in detection of oral precancerous and malignant lesions.

[Effects of small interfering RNA magnetic nanoparticles combination with external magnetic fields on survivin gene expression of bladder cancer cells and apoptosis].

OBJECTIVE: To investigate the effects of the small interfering RNA plasmid-dextran magnetic nanoparticles (siRNA-DMN) combination with external magnetic fields on silencing survivin gene expression of bladder cancer cells and apoptosis when DMN used as gene carrier to transfer siRNA-survivin recombinant plasmid in vivo. METHODS: The siRNA-survivin recombinant plasmid specific targeted survivin was synthesized in previous experiment. DMN were prepared by chemical coprecipitation method and used as gene carrier. The siRNA-DMN were constructed by static electricity of polylysine and transferred into human bladder cancer BIU-87 cells with the help of external magnetic fields. The growth inhibiting rate (IR) of BIU-87 cells was observed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide's test and the apoptosis index (AI) was detected by transferase-mediated dUTP nick end labeling method. The relatively transcription levels of survivin mRNA and protein expression were respectively detected by semi-quantitive reverse transcription polymerase chain reaction and Western Blotting techniques. RESULTS: The diameter, effective diameter and saturation magnetization of DMN-siRNA were about 10 - 12 nm, 94.8 nm and 0.19 emu/g, respectively. The IR (39.60%) and AI (28.72%), the relative expression of survivin mRNA and protein of siRNA-DMN combination with external magnetic fields on BIU-87 cells were significantly higher and lower than those in the control group and single siRNA-DMN group, respectively (P < 0.05). CONCLUSIONS: The siRNA-survivin plasmid-DMN combination with external magnetic fields could effectively inhibit survivin expression and induce BIU-87 cells apoptosis which provided experimental basis for the magnetic targeting gene therapy of bladder tumor.

[Effects of different immunodepressants on the sperm parameters of kidney transplant recipients].

OBJECTIVE: To study the effects of different immunodepressants on the sperm parameters of kidney transplant recipients. METHODS: In 15 healthy fertile men and 37 kidney transplant recipients, ejaculates were aseptically obtained by masturbation. Thirty-seven patients were divided into two groups, 20 patients were treated with Prograf (FK506) combination with mycophenolate mofetil (MMF) and prednisone; 17 patients were treated with cyclosporine (CsA) combination with azathioprine with prednisone. The sperm viability, mobility parameters such as prorsad percentage motility, straight line velocity (VSL), curve line velocity (VCL), velocity of average path (VAP) and morph were estimated with a computer-assisted sperm analyzer (CASA) provided with a multiple-exposure photography system. RESULTS: There were no significant difference in sperm viability rate [(81.7 +/- 5.7)%, (79.4 +/- 6.8)% and (83.8 +/- 6.0)%], VCL [(24.1 +/- 8.6)%, (23.9 +/- 4.4)%, (24.8 +/- 4.2)% ] and VAP [(19.7 +/- 6.6)%, (18.6 +/- 2.9)%, (21.0 +/- 4.0)%] among groups of FK506, CsA and control, respectively (P > 0.05). The rate of anomaly [(67.8 +/- 5.7)%], the prorsad percentage motility [(46.4 +/- 8.1)%] and VSL [(15.4 +/- 4.6)%] in the group of FK506 were respectively significantly lower and higher than those in the group of CsA [(80.1 +/- 5.6%, (33.3 +/- 6.4)%, (10.2 +/- 2.4)%] (P < 0.05). CONCLUSION: The application of FK506 combined with MMF could help recover the mobility and morphology of the sperm in kidney transplantation recipients.

A single nucleotide polymorphism in the matrix metalloproteinase-1 promoter enhances oral squamous cell carcinoma susceptibility in a Chinese population.

We genotyped 96 oral squamous cell carcinoma (OSCC) patients for the 1G/2G polymorphism of matrix metalloproteinase-1 (MMP-1) promoter -1607 bp using PCR-RFLP. A control population of 120 frequency-matched subjects was also genotyped for the same polymorphism. The detection frequency of 2G allele was significantly higher in OSCC subjects (76%) than in the control group (56.7%). The frequency of 2G allele had a significant difference between the OSCC and controls group (p = 0.00, Odds Ratio, OR = 2.232, 95% CI = 1.477-3.372). The genotype 2G/2G was found in 57.3% of the OSCC, and 34.2% in the controls. The proportion of 2G homozygote (2G/2G) was significantly higher in the OSCC group when compared to controls (p = 0.001, OR = 2.585, 95% CI = 1.487-4.494). OSCC patients were stratified by clinicopathological parameters including gender, smoking, clinical stage and lymph node metastasis, but the only statistically significant association with MMP-1 genotype was with smoking. The results showed that a SNP in the MMP-1 promoter -1607 bp was associated with OSCC susceptibility in a Chinese population.

[The effects of culture supernatant of bladder tumor cells on formation and function of dendritic cells].

AIM: To explore the effects of culture supernatant of bladder tumor cell lines BIU-87 on the development and function of peripheral blood mononuclear cell (PBMC)-derived dendritic cells. METHODS: The culture supernatant of BIU-87 cells was collected and used as the conditional culture medium for PBMC-derived DCs culture in experiment group. Normal cultured DCs were used as control group. The phenotypes of DCs in experiment group and in control group were analyzed by FACS. The capacity and difference to stimulate allogenetic T cells of PBMC-derived DCs from the two groups were evaluated by MTT colourimetry. RESULTS: Phenotypic analysis showed that DCs co-cultured with culture supernatant of BIU-87 cells expressed lower levels of CD1a, CD83 and CD86. Moreover, as compared with control group, the maturation of DCs was inhibited, and the capacity to stimulate allgentic T cell proliferation by DCs descended(P<0.05). CONCLUSION: The culture supernatant of BIU-87 cells can inhibit the development and function of DCs. It is a possible reason for the decrease of the number of DCs and the descent of its functional in the patients with bladder tumor.

[Effect of photodynamic therapy with metalloporphyrin compound on human prostate cancer PC-3 cells in vitro].

OBJECTIVE: To investigate the effect of the photodynamic therapy (PDT) with the new water-soluble metalloporphyrin compound on human prostate cancer PC-3 cells in vitro and the anticancer mechanism of PDT. METHODS: The new water-soluble manganese, 5,10,15, 20-tetra (N-methyl4-pyridyl) porphinato (2-) tetraiodide salt, was synthesized. The PC-3 cells were treated with the compound of serial concentrations(0, 0.1, 1, 1.0 micromol/L) followed by irradiation of different dosages of visible light. The techniques of MTT and Annexin-V/propidium iodide double-labeled flow cytometry (FCM) were applied to measuring the inhibitory effect of the compound on the growth activity and apoptosis of the cells. RESULTS: When the metalloporphyrin compound concentration was within 10 micromol/L and the irradiation time was within 30 min, the water-soluble metalloporphyrin compound had a significant inhibitory effect on the proliferation of PC-3 cells and induced PC-3 cell apoptosis, and the effects depended greatly on metalloporphyrin concentration and illumination dosages. Higher concentrations and dosages induced the death of the majority of PC-3 cells. CONCLUSION: The PDT of the water-soluble metalloporphyrin compound followed by light irradiation has a distinctive killing effect on PC-3 cells in vitro, and the rates of proliferation inhibition and cell apoptosis are correlated with metalloporphyrin concentration and the dosages of light irradiation. The results suggest that the mechanism of metalloporphyrin PDT may be involved with the induction of apoptosis in human prostate cancer cells.

[Preparation and feasibility of superparamagnetic dextran iron oxide nanoparticles as gene carrier].

BACKGROUND & OBJECTIVE: Application of magnetic nano- particles as gene carrier in gene therapy of tumor has developed quickly. To obtain a new type non-viral gene introduction and therapy system,which is convenient,and can drive target gene to express highly and stably,this study was designed to explore the preparation of superparamagnetic dextran iron oxide nanoparticles(SDION),and the feasibility of SDION used as gene carrier in vitro. METHODS: SDION were prepared by chemical co-precipitation,separated by gel filtration chromatography on Sephacryl S-300HR,and centrifugation techniques,characterized by transmission electron microscopy,laser scattering system,and vibrating sample magnetometer signal processor. The green fluorescent protein-C2 (GFP-C2) plasmid was used as target gene. SDION-GFP- C2 compounds were synthesized by oxidation-reduction reaction. The connection rate of SDION and GFP-C2 was analyzed by agarose electrophoresis,and evaluated by measuring concentration of GFP in the supernatant after centrifugation. Liposome transfection was used as control,the efficiencies of SDION and liposome in transferring GFP gene into human bladder cancer BIU-87 cells were evaluated under fluorescence microscope in vitro. RESULTS: The diameter of SDION ranged from 3 nm to 8 nm, the effective diameter was 59.2 nm, and the saturation magnetization was 0.23 emu/g. After oxidized by sodium periodate of 10 mmol/L,and deoxidized by sodium hydride boron of 0.5 mol/L, SDION could connect with GFP in maximum degree,the transfection efficiency of SDION as gene carrier was about 45%, even higher than that of liposome (about 30%). CONCLUSION: SDION could connect with GFP plasmid by oxidation- reduction reaction,and success to transfer GFP gene into human bladder cancer BIU-87 cells in vitro.

[The effects of the active constituents of Alisma orientalis on renal stone formation and bikunin expression in rat urolithiasis model].

OBJECTIVE: To evaluate the effects of the active constituents of Alisma orientalis on the expression of bikunin mRNA in rat urolithiasis model, and explore the mechanism of this traditional Chinese medicine on prevention of urinary calculi. METHODS: Modern phytochemistry and bioactivity guided isolation techniques were applied to extract the active constituents of Alisma orientalis. Hyperoxaluria and the renal oxalate calcium stone formation were induced in rats by infusion into the stomach with 1% ethylene glycol and 2% ammonium chloride. 30 adult male Wistar rats were randomized into 3 groups of 10 rats: control group, infused into the stomach with running water; stone-forming group, infused into the stomach with 1% ethylene glycol and 2% ammonium chloride so as to make into renal oxalate calcium stone model; and group of Alisma orientalis, infused into the stomach with 2% ammonium chloride and the constituents of Alisma orientalis. Four weeks after the rats were killed and their kidneys were taken out. Reverse transcription polymerase chain reaction technique was used to examine the bikunin mRNA expression levels in the rat renal tissues. The calcium oxalate deposits in the kidneys were detected by microscopy. The serum creatinnine and blood urea nitrogen levels, renal tissue calcium content, 24 h urinary calcium and oxalate excretion were also detected. RESULTS: In the group administered with the active constituents of Alisma orientalis, calcium oxalate deposits in the kidney, serum creatinnine and blood urea nitrogen levels, the bikunin mRNA expression levels, renal tissue calcium content and 24 h urinary calcium excretion were all significantly lower than those in the model group (the bikunin mRNA expression levels: 0.53 +/- 0.17 vs 0.71 +/- 0.25, P < 0.05; renal tissue calcium content: 4.70 mg/g +/- 0.08 mg/g vs 9.49 mg/g +/- 0.45 mg/g, P < 0.01; 24 h urinary calcium excretion: 37 micromol +/- 2 micromol vs 62 micromol +/- 2 micromol, P < 0.01). CONCLUSION: The active constituents of Alisma orientalis can down-regulate the bikunin mRNA expression, decrease the calcium oxalate formation in rat kidney, and inhibit the renal stone formation in rat urolithiasis model.

[Effects of baicalin on the expression of pro-MMP-1 and MMP-3 in human gingival fibroblasts and periodontal ligament cells].

OBJECTIVE: To investigate the influence of baicalin on the IL-1beta induced pro-MMP-1 in HGF and the effects of baicalin on MMP-3 expression in periodontal ligament cells (PDLCs). METHODS: The amount of secreted pro-MMP-1 and MMP-3 expression was detected by ELISA and cell immunochemistry. RESULTS: (1) The amount of secreted pro-MMP-1 (3.333 +/- 0.123) microg/L increased significantly following 1 microg/L of IL-1beta, compared with control group (1.960 +/- 0.180) microg/L. Addition of baicalin to cell culture medium for 1 hour following IL-1beta decreased pro-MMP-1 secretion in a dose-dependent manner in the range of 10 approximately 1,000 microg/L. (2) 1 microg/L IL-1beta could significantly stimulate the synthesis and secretion of MMP-3 in PDLCs. (3) The baicalin could not interfere the synthesis of MMP-3, but could inhibit the release of MMP-3 from PDLCs. CONCLUSIONS: Baicalin could inhibit the secretion of pro-MMP-1 and MMP-3 expression in IL-1beta induced HGF and PDLCs, which suggests that baicalin may play an important role in preventing and treating periodontal disease.

Influences of dibutyryl cyclic adenosine monophosphate and forskolin on human sperm motility in vitro.

AIM: To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro. METHODS: Semen samples, aseptically obtained by masturbation and prepared by swim-up technique from 20 fertile men, were incubated with different concentrations of dbcAMP and forskolin at 37 deg. Measurements were carried out after 10 min, 20 min, 30 min and 60 min incubation. Motility parameters were estimated by using an automatic analyzing system. RESULTS: Treatment with dbcAMP or forskolin resulted in a significant increase in sperm motility and progressive motility. The larger the concentrations of dbcAMP or forskolin, the greater the effect appeared. The straight linear velocity and curvilinear velocity were not affected by both agents. CONCLUSION: dbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro.

[An experimental study of effect of different extracts of Alisma orientalis on urinary calcium oxalate stones formation in rats].

OBJECTIVE: To study the effect of different extracts of Alisma orientalis on urinary calcium oxalate stone formation in rats and to identify the effective constituents. METHOD: Different extracts were administered through a stomach tube to rats of different groups with renal calcium oxalate stones induced by ethylene glycol (EG) and ammonium chloride (AC). RESULT: In the rats administered with ethyl acetate elution of ethyl acetate extract, blood Cr, BUN, renal tissue calcium content, urinary calcium excretion and crystals deposition in renal tissue were significantly lower than those of the stone formation group. CONCLUSION: The ethyl acetate elution of ethyl acetate fraction extract of Alisma orientalis can significantly inhibit urinary calcium oxalate stone formation in rats and be the most effective constituent of Alisma orientalis.

[Biological characteristics of rabbit corporal smooth muscle cells cultured in vitro].

OBJECTIVES: To study the biological characteristics of rabbit corporal smooth muscle cells (SMC) cultured in vitro. METHODS: In vitro tissue culture technique, morphological observation, cell counting, mitosis index and adhesion rate evaluation were applied to study the biological features of the SMC. RESULTS: 1. SMC were spindle-shaped and parallel along their longitudinal axis, showing obvious orientation. 2. The attachment and the proliferation of SMC in vitro were rapid. SMC cultured in vitro can grow and maintain their steady characteristics provided appropriate passage rate and culture condition. CONCLUSIONS: The SMC cultured in vitro are proved to be used to evaluate and investigate the effect of some medicine on penile erection.

Influence of several uropathogenic microorganisms on human sperm motility parameters in vitro.

AIM: The effects of certain uropathogenic microorganisms (Neisseria gonorrhoeae, Staphylococcus aureus, Staphylococcus epidermidis and Mycobacterium tuberculosis) on human sperm motility characteristics were studied in vitro. METHODS: In 10 healthy fertile men, ejaculates were aseptically obtained by masturbation and with a swim-up technique, a sperm suspension of high motility and purity was obtained. Several uropathogenic bacteria were obtained from outpatients with genitourinary tract infections. The sperm suspension was incubated with the pathogens at a bacteria: sperm ratio of 50:1 at 37deg. The sperm mobility parameters were estimated with a computer-assisted sperm analyzer (CASA) provided with a multiple-exposure photography system (Madi Corp., Zhejiang, China). Measurements were carried out at 0, 2 and 4 hours of incubation. RESULTS: Staphylococcus aureus significantly decreased the sperm motility and viability, but Staphylococcus epidermidis, Mycobacterium tuberculosis and Neisseria gonorrhoeae did not. CONCLUSION: Staphylococcus aureus has an inhibitory effect on human sperm motility in vitro.