10.3969/j.issn.2095-4344.2013.24.024

Objective To study the curative effect of transurethral incision for male urethral stricture or atresia. Methods A retrospective) review was made on 55 male patients with urethral stricture or atresia treated by transurethral incision. Results The success rate of the operation on one session was 90.9% (50/55), while the remaining 5 patients were cured by two times of operations.Forty-seven patients were followed for 6~12 months (mean,10 months).Urethral dilatation was performed for once within 1 week after the remorval of catheter in 10 patients,for 3~5 times within 3 months after operation in 21 patients,and after the third postoperative month in 16 patients. Conclusions Transurethral incision for male urethral stricture or atresia is effective.

Objective To explore the effects of quercetin and quercetin in combination with cisplatin on adhesion, migration and invasion of HeLa cells. Methods Adhesion, migration and invasion of HeLa cells treated with quercetin and quercetin in combination with cisplatin were measured by adhesion assay, wound healing assay, and transwell chamber method respectively. Results The results showed that quercetin and quercetin in combination with cisplatin could inhibit adhesion, migration and invasion of HeLa cells. The adhesive ratio, migration rate and the invasiveness were negatively proportional to concentration of quercetin and quercetin in combination with cisplatin. With increasing concentration of quercetin from 20 to 80 μmol/L, the adhesive ratio decreased from ( 82. 2±1. 5 ) % to ( 48. 4±1. 1 ) % ; the migration rote decreased from (7.26±0. 20) μm/h to (3.78±0. 64) μm/h; the invasiveness decreased from ( 124. 3± 1. 5) piece to (90. 7±2. 1 ) piece(P <0. 05). In quercetin and quereetin in combination with 10 μmol/L cisplatin treatment group, with quercetin concentration increasing from 20 to 80 μmol/L, the adhesive ratio decreased from (42. 6±1. 2) % to ( 27. 5±1. 7 ) %, the migration rate decreased from ( 2. 20±0. 33) μm/h to (0.72±0. 19) μm/h and the invasiveness decreased from (78.7±2.5) piece to (44.0±4.0) piece (P < 0. 05 ). Compared with the quercetin groups, querectin in combination with cisplatin groups had significantly higher inhibitory effects ( P < 0. 05 ). Conclusions Quercetin and quercetin in combination with cisplatin can inhibit adhesion and migration and invasion of HeLa cells. Quercetin can enhance the inhibitory effect of cisplatin on HeLa cell adhesion, migration and invasion.

Objective To investigate the expression and significance of the human telomeruse gene (hTERC)in the cytologic specimens of cervix for early cervical lesion screening.Methods Cytologic samples of cervix including 120 normal persons,86 CIN patients and 14 SCC patients were analyzed for aberrations of 3q26 using a commercially available two-color FISH probe.Results Positive expression rates of hTERC gene in normal,CIN Ⅰ ,CIN Ⅱ,CIN Ⅲ and SCC samples were 1.7%(2/120),2.6%(1/38),55.6%(10/118),87.5%(14/16),100.0%(14/14),100.0%(14/14),respectively.Compared with CIN Ⅰ/CIN Ⅱ group,hTERC gene positive expressions in SCC/CIN Ⅲ group were higher significantly(P=0.01),and them were also difference between CIN Ⅰ-CIN Ⅲ group and normal controls(X~2=113.52,P ＜0.01).With progression from CIN to carcinoma,polyploidization resulting in aneuploidy and high TERC gene copy numbers in SCC group ascended significantly.Among the total abnormal cells,the number of the cells percentage which hTERC amplification signals was more than 3:3 in CIN Ⅰ ,CIN Ⅱ,CIN Ⅲ and SCC groups were 6.12%,7.98%,28.07% and 33.97%,respectively.Condusion The hTERC gene is increasingly gained with progression of CIN and may appear to be another screening testmarker for early cervical cancer screening and accessory diagnosis.

Objective The purpose of this study was to investigate the mechanism of action of polypeptide extracts of Eupolyphaga sinensis Walker ( ESW) against oxidative aging.Methods Mice were intraperitoneally injected D-galac-tose for consecutive 20 days to establish an aging mouse model.The model mice were administered with different doses of ESW polypeptide (0, 40, 80, 160 mg/kg/d).The normal activity, movement and anti-stress ability of the mice were ob-served.The activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX) in blood and different tissues and the content of glutathione ( GSH) and malondialdehyde ( MDA) of the aging mice were assessed by xanthin oxidase activity measurement and spectrophotometry, respectively.The expression of nuclear factor erythroid 2-re-lated factor 2 (Nrf2) in Caco-2 cells was detected by immunofluorescence.Results Comparing the control and polypep-tide groups, there were significant decreases of body weight gain, organ indexes, anti-stress ability and activity capacity, the activity of SOD, CAT, GSH-PX and the content of GSH, and an increase of the content of MDA in blood and different tissues in the aging mice.With the increasing dose of polypeptide extracts of ESW, the body weight gain, organ indexes of the liver, spleen and kidney were significantly increased, the static and dynamic exercise time was prolonged in the poly-peptide group, and their abilities of hypoxia tolerance and heat tolerance were close to that of normal controls.The SOD, CAT, GSH-PX activity and GSH level in blood and different tissues were significantly increased, but MDA content de-creased.The expression of Nrf2 in Caco-2 cell nuclei was significantly increased in the polypeptide group, close to that of the positive control group.Conclusions The results of our study show that polypeptide extracts of ESW improve the anti-stress and antioxidative capacity in D-galactose-induced mouse models of oxidative aging by initiating Nrf2-ARE antioxidant signaling pathway, therefore, delay the oxidative aging in mice.

Objective To investigate the pathogenicity of Alternaria alternata in ICR mice. Methods The cell suspension of Alternaria alternata was inoculated into testes and footpads of normal and immunocompromised ICR mice. H istopathologic examination and fungal culture were performed on 5th d, 7th d, 10 th d, 14th d, 21st d, 28th d, 35th d and 42nd d respectively after inoculation. Results The abscesses were developed at the inoculated sites in all normal and immunocompromised mice. The lesions in immunocompromised mice were more severe and persisted longer than those in normal mice. The abscesses in footpads mostl y disappeared within 14 days in normal mice but in 28 to 35 days in immunocompro mised mice. Alternaria alternata was isolated from the inoculated tissues in bot h groups. The histopathologic examination showed abscess and granulomas in which thick hyphae were observed. The necropsy revealed no disseminated infection of Alternaria alternate in all the test mice. Conclusion Alternaria alternata is a kind of opportunistic fungus with low virulence.

Objective To evaluate the feasibility of bladder extracellular matrix (BACM) seeding with autologous cultured vaginal smooth muscle cells (VSMC) repaired rabbit vaginal defect.Methods This study included 24 female rabbits.BACM and vaginal acellular matrix (VACM) were obtained from 8 rabbits by decellularization process.The other 16 rabbits were randomly divided into experimental and control groups.Vaginal tissue biopsies ( ～ 1 cm2) were harvested from female New Zealand rabbits.VSMC were cultured and stained with a-smooth muscle actin antibodies. Cultured VSMC cells were seeded on BACM ( experimental group) or VACM ( control group) at a cell density of 1 × 107 cells/cm2.The cell-seeded matrixes were cultured for 5days.In experimental group of 8 rabbits,a 2 cm segment of vagina was resected and replaced with BACM seeding with VSMC.Then the regenerative segment was studied with histological technique by hematoxylin-eosin staining after 3,6 and 12 weeks postoperative aud vaginography was performed at 12 weeks postoperative.The 8 rabbits in control group underwent the exact same procedure as above but the vaginal defect was repaired with VACM seeding with VSMC,Results The prepared BACM and VACM were transparent,HE staining and scanning electron microscopy showed the two acellular matrix was both consisted of abundant network of fibers with the regular arrangement,without cellular debris.Primary culture of VSMC was successfully established and passaged,and was uniformly spindle-shaped in the confluent state and showed a characteristic hill and valley'formation.Immunohistochemical staining showed VSMC in culture stained positively with a-smooth muscle actin antibodies.VSMC began to adhere to the BACM 5 hour after implanting and the number of the adhered cells increased with time.Cells gradually expanded and showed the typical morphology of smooth muscle cells.Three weeks after implantation in vivo,the luminal surface of matrix was completely covered by vaginal epithelial tissue,a layer of smooth muscle cells was formed in the outer surface of the matrix.Multilayered vaginal epithelial and improved development of organized muscle bundles was observed after 6 weeks.The regenerative tissue was equivalent to the normal vaginal tissue at 12 weeks postoperatively.Vaginography demonstrated the maintenance of full patency and a wide vaginal caliber without fibrosis and graft rejection.There was no significant difference in all evaluated items between experimental and control groups.Conclusions BACM has the same regenerative process as VACM in the replacement of vaginal defect.Moreover,BACM has wider source and appears to be an suitable material for vaginal replacement.

Objective To study the mechanism of adaptive immunity against Rhizopus arrihizus (R. arrihizus) infections. Methods Bone marrow derived dendritic cells (BMDCs) were separated from C57BL/6 mice and Card9-/- mice and then were cultured in vitro. Resting spores and swollen spores of R. arrihizus were in vitro co-cultured with BMDCs with or without Syk inhibition. Secretion of cytokines ( IL-23, IL-1βand IL-12) was analyzed by ELISA after 24 hours of culture. Na?ve T cells derived from C57BL/6 mice were in vitro co-cultured with spore-stimulated BMDCs for four days. Levels of IL-17A and IFN-γ in supernatants of cell culture were analyzed by ELISA. Flow cytometry was performed to analyze T cell differ-entiation. Confocal microscopy was used to observe the images of stained β-glucan on the surface of resting and swollen spores. Swollen spores were co-cultured with Dectin-1, Dectin-2, TLR2 and mannose receptor ( MMR) , and the binding results were analyzed by flow cytometry. Results Swollen spores but resting spores could induce the maturation of BMDCs and promote the secretion of cytokines (IL-23, IL-1βand IL-12). Co-culturing T cells with swollen spore-stimulated BMDCs enhanced their differentiation to Th17 and Th1. In addition, swollen spores promoted the secretion of Th1-related cytokine ( IFN-γ) and Th17-related cytokine (IL-17A). Adding Syk inhibitor to Card9-/-BMDCs or wild type BMDCs significantly inhibited the secretion of cytokines and T cell differentiation, especially in the Card9-/- group. β-glucan was overserved on the surface of swollen spores, but not on resting spores. On the surface of swollen spores existed pathogen associated molecular patterns ( PAMPs) that could bind with Dectin-1 and TLR2. Conclusion Swollen spores of R. arrihizus could active BMDCs to secrete cytokines of IL-23, IL-1β and IL-12 and trigger T cell responses in vitro. The possible mechanism might be associated with β-glucan exposed on the surface of swollen spores that binds with Dectin-1. The responses between BMDCs and R. arrihizus are Syk-Card9-dependent.

OBJECTIVEThe purpose of this study is to construct a pyruvate oxidase gene deficiency variant strain of Streptococcus oralis (S. oralis).

METHODSThe sopox gene, which was got using polymerase chain reaction (PCR), and the 130-basepair segment of which was cut down with endonuclease BamHI, and transferred into S. oralis (ATCC10557) by using electrotransformation. The authors obtained a variant strain of S. oralis, and then the catalase activity of the first culture and 3-4 subcultures was examined.

RESULTSThe authors obtained a pyruvate oxidase gene deficiency variant strain of S. orlis. The catalase activity examination showed that the ability of producing H2 O2 of the variant strain of S. orlis declined, whose catalase activity was between those of the positive control (ATCC10557) and the negative control (Escherichia coli, JM109). But the produced H2 O2 quantity of their subcultures was less than that of the negative control.

CONCLUSIONThe construction of the pyruvate oxidase gene deficiency variant strain of Streptococcus oralis is successful.

Cloning, Molecular ; Genes, Bacterial ; Genetic Engineering ; Genetic Variation ; Polymerase Chain Reaction ; Pyruvate Oxidase ; genetics ; Streptococcus oralis ; enzymology ; genetics

BACKGROUND:It has been proved that erythropoietin can promotes angiogenesis in injured tissue, which is closely related to the proliferation and differentiation of endothelial progenitor cel s. However, the involved mechanism remains unclear yet.
OBJECTIVE:To investigate the effect of erythropoietin on the function and activity of bone marrow-derived endothelial progenitor cel s in mice, and to explore the signal pathway.
METHODS:The endothelial progenitor cel s from the bone marrow of mice were separated by means of density gradient centrifugation and then cultured. The cel s were preconditioned by specific inhibitor of PI3K
(LY294002), and were divided into the fol owing groups:EGM-2 group, three erythropoietin preconditioned groups (the concentrations of erythropoietin in medium were 1, 5, 10 U/mL respectively), erythropoietin+LY group (10 U/mL erythropoietin and 10 mmol/L LY294002 in medium), LY group (10 mmol/L LY294002 in medium), dimethyl sulfoxide group (1 mL/L dimethyl sulfoxide in medium). The cel proliferation and apoptosis were evaluated by cel counting kit-8 and flow cytometry respectively. The contents of endothelial nitric oxide synthase and vascular endothelial growth factor in cel lysates were detected by the method of ELISA, and the expressions of Akt and p-Akt were by western blot assay.
RESULTS AND CONCLUSION:Erythropoietin could promote the proliferation of endothelial progenitor cel s in a dose-dependent manner, which was, however, completely inhibited by LY294002. The apoptosis rate in the erythropoietin preconditioned groups was significantly lower than that in the erythropoietin+LY group. The contents of endothelial nitric oxide synthase and vascular endothelial growth factor in cel lysates of LY group and erythropoietin+LY group were significantly lower than those in the erythropoietin groups. There was no difference in Akt expression found in each group, while the p-Akt expression in the erythropoietin+LY group was significantly lower than that in the erythropoietin groups. The above results reveal that erythropoietin can promote the proliferation of endothelial progenitor cel s and decrease the cel apoptosis, which is depending on PI3K/Akt signal pathway.