0.05).Urethroscopy showed a smooth and intact internal mucosa,wide urethral caliber and normal-appearing urethral tissue in the 2 groups. Conclusions VECM has the same regenerative process as UECM in the replacement of urethral defect.Moreover,VECM has wider source and better elasticity and mechanical properties,so VECM appears to be an ideal material for urethral replacement.

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

OBJECTIVE To assess the outcomesof three different therapeutic approaches for the treatment of T3N0M0(stage Ⅲ) glottic squamous cell carcinoma. METHODS Sixty-five cases of T3N0M0 glottic squamous cell carcinoma treated with curative intent by three different modalities include vertical hemilaryngectomy(VHL,n=21),total laryngectomy(TL,n=31)and radiotherapy(RT,n=13) were reviewed retrospectively. The survival rate,recurrence rate at the primary lesion site and jugular lymph node, and laryngeal preservation was compared among three methods. RESULTS There was no significant difference in the overall survival rates(Cox proportion hazard model) and recurrence rates at the primary lesion site or jugular lymph node among the three methods. Laryngeal function was preserved in 100% of the cases in the VHL and RT groups after initial treatment. CONCLUSION The three treatment modalities had statistically similar survival and recurrence rates. Patients treated with VHL and RT had a higher rate of laryngeal preservation compared to that of TL, hence VHL or RT is a valid alternative to TL in treating selected patients with T3N0M0 glottic squamous cell carcinoma.

Objective To evaluate the clinical characters, management and prognostic factors of patients with differentiated invasive thyroid carcinoma (DITC). Methods The data were analyzed retrospectively for 114 DITC patients treated at Department of Head and Neck Surgery of Sun Yat-sen University Cancer Center. Survival analysis was performed by Kaplan-Meier method, comparison among/between groups was performed using log-rank test, and multivariate analysis was carried out using Cox proportional hazard model. Results After surgery, 68 patients were with tumor residue. The 5-year and 10-year overall survival rate were 91.9% and 80.1% respectively in all patients, while the 10-year overall survival rate were 88.5% 、78.5% and 53.1% in no tumor residue group, micro-residue group and grossresidue group respectively. This study failed to prove that radiotherapy might improve the survival rate in patients with postoperative tumor residue. Multivariate analysis indicated that age, invasion to esophagus and recurrence predict the prognosis. Conclusion DITC may be treated mainly by surgical operation. Radical resection is the key factor in the treatment of DITC. Patients with DITC have a relatively poor prognosis.Age, esophagus invasion and status of tumor residue are the most important factors affecting the prognosis.

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

Objective To investigate the prognostic factors of differentiated thyroid carcinoma (DTC) in patients younger than 45 years. Methods Clinical data of all patients less than 45 years old at presentation with DTC in our hospital from Jan 1985 to Dec 1997 were analyzed retrospectively. Single variable analysis was performed by life-table method. Multivariate analysis was performed by Cox proportional hazard model. Results Two hundred and seventy two cases were analyzed. The overall 10-year survival rate was 93. 0%. The main prognostic factors influencing survival were age at presentation, the status of lymph node metastasis and distant metastasis; distant metastasis was the risk factor independently influencing survival by multivariate analysis. Conclusions Distant metastasis is the factor influencing survival significantly. The prognosis of these patients without distant metastasis is good. Total or near-total thyroidectomy and postoperative 131I therapy may be essential for a better prognosis in patients with distant metastasis.

ObjectiveTo study the value of the exfoliative cytology of prostatic fluid obtained from patients with elevated PSA in prostate cancer diagnosis.MethodsProstatic fluid was obtained from 130 patients with elevated PSA before prostate biopsy and then the Wright's stain and cytological class were done.Each cytological class and patient's age,PSA,total prostate volume,prostatic fluid volume and the number of leukocyte in the prostatic fluid were recorded.The relationship of leukocyte number and patient's age,PSA and prostate volume was analyzed by Spearman correlation test.ResultsProstate biopsy pathology results showed that there were 77 (59.2％) cancer cases and 53 (40.8％) non-cancer cases.Patient numbers in cytological class 1 to 5 were 28 (21.5％),32 (24.6％),22 (16.9％),36 (27.7％),12(9.2％),respectively.The prostate fluid cytology had a specificity of 100％ and high sensitivity of 62.5％(10/16) in patients with PSA≥20 μg/L.PSA value had significant difference between class 1,2,3,4and 5.Significant correlation was found among the prostatic fluid volume,total leukocyte number and prostate volume.Prostate volume,leukycyte density and total leukycyte number was significant higher in noncancer patients than in prostate cancer patients.ConclusionsThe exfoliative cytology of prostatic fluid is a valuable method in detecting prostate cancer,particularly in patients with high PSA levels.It has the advantages of non-invasion and less injury than prostate biopsy.There is a relationship between elevated PSA value and high leukocyte numbers.

Objective To investigate the distribution of Human papillomavirus(HPV)genotyping and age in women treated in People′s Hospital of Hubei Province.At the same time,we want to analyse the characteristics of HPV infection.Methods HPV genotyping was detected from the cervical exfoliated cell samples from 7 746 female patients treated in People′s Hospital of Hubei Province from January 6,2016 to January 12,2017.Results There were 1 336 positive cases detected in 7 746 cases of female pa-tients,and the total positive rate was 17.2%.The top eight of HPV subtype infection were 16,52,58,53,81,18,39,56.There were 1 076 cases of single infection,accounting for 80.5%,and the infection of the two types were found in 202 cases,accounting for 15.1%.There were 47 cases of triple infections,accounting for 3.5% and fourfold infection and more were in 11 cases,accounting for 0.9% of all cases.The infection rate of HPV and the infection rate of middle and high risk subtype increased with age,and the infection rate of low risk subtype decreased with age.The detection rate of HPV infection in different age groups had a statistically significant difference(P<0.05).Conclusion The survey of female HPV infection subtype in Hubei Province conforms to the Asian population distribution,and the HPV high-risk subtype infection is mainly distributed in the population over the age of 40.

OBJECTIVE: The current study was designed to examine the expression of Skp2 gene in laryngeal squamous cell carcinoma (LSCC) and to investigate the role of Skp2 gene in tumorigenesis and progression of LSCC. METHOD: FQ-PCR method was used to examined the expression of Skp2 gene in 40 LSCC and 10 normal laryngeal mucosa tissues, and relationship between its expression and clinical biological factors of patients with LSCC was analyzed. RESULT: The median copy number of Skp2 mRNA expression in LSCC was 6622.54 copy/microg RNA, the median copy number of Skp2 mRNA expression in normal laryngeal mucosa tissues was 0 copy/microg RNA, there was a very significant difference between them (P < 0.01); The positive rate of Skp2 mRNA expression in LSCC and adjacent normal laryngeal tissue were 50%, 0, respectively (P < 0.01). The median copy number of Skp2 RNA expression in LSCC with cervical lymph node metastasis was 617138.4 copy/microg RNA, the median copy number of Skp2 mRNA expression in LSCC without cervical lymph node metastasis was 0 copy/microg RNA, there was a very significant difference between them (P < 0.05); The positive rate of Skp2 mRNA expression in LSCC with and without cervical lymph node metastasis were 100.00%, 35.48%, respectively (P < 0.01). CONCLUSION Skp2 gene might have relation with the cervical lymph node metastasis of LSCC. FQ-PCR is an accurate assay to detecting expression of Skp2 mRNA in patient with LSCC. The level of Skp2 mRNA expression might be a new and more accurate marker, and it can be used to predict cervical lymph node metastasis of LSCC.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; genetics ; Female ; Gene Expression ; Humans ; Laryngeal Neoplasms ; genetics ; Lymphatic Metastasis ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; RNA, Messenger ; genetics ; S-Phase Kinase-Associated Proteins ; genetics ; metabolism

The purpose of this study is to evaluate theretention capabilities of poloxamer-basedhydrogels for vaginal application with nonoxinol-9 as the model drug. Twohydrogel formulations, which contained 18% poloxamer 407 plus 1% poloxamer 188 (GEL1, relative hydrophobic) or 6% poloxamer 188 (GEL2, relative hydrophilic), were compared with respect to the rheological properties,hydrogel erosion and drug release. The vaginal retention capabilities of these hydrogel formulations were further determined in two small animal models, including drug quantitation of vaginal rinsing fluid in mice and isotope tracing withTc in rats. The two formulations exhibited similar phase transition temperatures ranging from 27 to 32 °C. Increasing the content of poloxamer 188 resulted in higher rheological moduli under body temperature, but slightly accelerated hydrogel erosion and drug release. When compared, GEL1 was eliminated significantly slower in rat vagina than GEL2, while the vaginal retention of these two hydrogel formulations behaved similarly in mice. In conclusion, increases in the hydrophilic content of formulations led to faster hydrogel erosion, drug release and intravaginal elimination. Rats appear to be a better animal model than mice to evaluate thehydrogel for vaginal application.