Dentin matrix protein 1 and phosphate homeostasis are critical for postnatal pulp, dentin and enamel formation / 国际口腔科学杂志·英文版

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).

Holmium laser urethrotomy for male urethral stricture / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate transurethral holmium laser incision, its safety and effect in the treatment of male urethral stricture.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>Holmium laser urethrotomy is a safe, effective and minimally invasive therapeutic modality for male urethral stricture.</p>

Protective effect of baicalin on experimental periodontitis in rats and its possible mechanisms / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of baicalin on the experimental periodontitis in rats, as well as the expression of MMP-1, MMP-2, MMP-9.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Expression of connexin43 and its correlation with the expression of bcl-2 and bax protein in transitional cell carcinoma of the bladder / 肿瘤

Objective: To study the expression of connexin43 (Cx43) gene and its correlation with the expression of apoptosis related genes bcl-2 and bax in transitional cell carcinoma of the bladder (BTCC), and to investigate the role of Cx43 in the BTCC. Methods: Reverse transcription polymerase chain reaction was used to detect the expression of Cx43 mRNA and immunocytochemistry technique was used to detect the expression of bcl-2 and bax proteins in 60 cases of BTCC tissue, and compared with that of 15 cases of pericancerous tissue and 15 cases of normal bladder tissue. Results: The positive rate of Cx43 mRNA expression in 60 cases of BTCC tissues was 43.33% which was significantly lower than that in pericancerous tissues (73.33% ) and normal tissues (100%) (x2 = 17.58, P 0.05). Expression of Cx43 negatively correlated with bcl-2 protein (r = 0.63, P < 0.01) and positively correlated with bax protein (r = 0.52, P < 0.01). Conclusion: The down-regulated expression of Cx43 gene was closely associated with the development, invasion and metastasis of BTCC. It could be used a prognostic indicator for BTCC. Cx43 gene may have antagonistic effects with bcl-2 gene and synergistic effects with bax gene in the initiation and progression of BTCC.

Small interfering RNA inhibits the expression of connexin43 in the human corpus cavernosum penis smooth muscle cells / 中华男科学杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Effects of different immunodepressants on the sperm parameters of kidney transplant recipients / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the effects of different immunodepressants on the sperm parameters of kidney transplant recipients.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>The application of FK506 combined with MMF could help recover the mobility and morphology of the sperm in kidney transplantation recipients.</p>

Effects of small interfering RNA magnetic nanoparticles combination with external magnetic fields on survivin gene expression of bladder cancer cells and apoptosis / 中华外科杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSIONS</b>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.</p>

Effect of photodynamic therapy with metalloporphyrin compound on human prostate cancer PC-3 cells in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Effects of baicalin on the expression of pro-MMP-1 and MMP-3 in human gingival fibroblasts and periodontal ligament cells / 中华口腔医学杂志

<p><b>OBJECTIVE</b>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).</p><p><b>METHODS</b>The amount of secreted pro-MMP-1 and MMP-3 expression was detected by ELISA and cell immunochemistry.</p><p><b>RESULTS</b>(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.</p><p><b>CONCLUSIONS</b>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.</p>

Influences of dibutyryl cyclic adenosine monophosphate and forskolin on human sperm motility in vitro / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>dbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro.</p>

An experimental study of effect of different extracts of Alisma orientalis on urinary calcium oxalate stones formation in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of different extracts of Alisma orientalis on urinary calcium oxalate stone formation in rats and to identify the effective constituents.</p><p><b>METHOD</b>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).</p><p><b>RESULT</b>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.</p><p><b>CONCLUSION</b>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.</p>

Biological characteristics of rabbit corporal smooth muscle cells cultured in vitro / 中华男科学杂志

<p><b>OBJECTIVES</b>To study the biological characteristics of rabbit corporal smooth muscle cells (SMC) cultured in vitro.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>The SMC cultured in vitro are proved to be used to evaluate and investigate the effect of some medicine on penile erection.</p>

Influence of several uropathogenic microorganisms on human sperm motility parameters in vitro / 亚洲男科学杂志(英文版)

<p><b>AIM</b>The effects of certain uropathogenic microorganisms (Neisseria gonorrhoeae, Staphylococcus aureus, Staphylococcus epidermidis and Mycobacterium tuberculosis) on human sperm motility characteristics were studied in vitro.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>Staphylococcus aureus significantly decreased the sperm motility and viability, but Staphylococcus epidermidis, Mycobacterium tuberculosis and Neisseria gonorrhoeae did not.</p><p><b>CONCLUSION</b>Staphylococcus aureus has an inhibitory effect on human sperm motility in vitro.</p>