BACKGROUND:Anticancer drug deguelin delays the embryonic development of zebrafish, but its exact mechanism is not yet clear.
OBJECTIVE:To explore expression differences in genes and signaling pathways in deguelin-treated zebrafish embryos.
METHODS:Zebrafish embryos were harvested from zebrafish fed according to the Zebrafish Book. Zebrafish embryos at 2-cel stage were selected and divided into two groups:control group treated with dimethyl sulfoxide incubator liquid, and experimental group treated with 0.6μmol/L deguelin. Total RNA was extracted from the two groups at sphere stage. Then, gene chip technique was used to detect differential y expressed genes in the deguelin-treated zebrafish embryos. Real-time quantitative PCR was employed to validate microarray cluster analysis and pathway analysis to explore the mechanism of action of deguelin.
RESULTS AND CONCLUSION:Chip results showed that 407 genes were upregulated more than three times and 461 genes were downregulated more than three times after deguelin treatment. PCR validation results were consistent with those of the chip. Fourteen pathways were identified by KEGG pathway analysis. Deguelin may play an important role by intervening cel metabolism growth and differentiation.

ObjectiveTo report 18 cases of Bowen dis ea se and erythroplasia of Queyrat.MethodsClinicopatholog ical data of 18 cases of Bowen disease and erythroplasia of Queyrat were present ed.10 cases (mean age 49 year)underwent circumcision and local resection and 8 c ases electric coagulation.Results15 cases have been fo llowed up for 4～48 months.2 cases had new disease at an untreated area and the o ther 13 have been free of the disease.Recurrence ocurred in 2 were cured with 5% 5-fluorouracil cream external application.ConclusionsThe result of the surgical treatment for Bowen disease and erythroplasia of Qu eyrat is good and an alternative therapy with 5-fluorouracil for external appli cation may be considered.Adequate biopsy including deeper tissue is necessary fo r differential diagnosis between carcinoma in situ and invasive penile carcinoma or a variety of lesions of the male genitalia.

Objective To study the clinical and pathological features of a case of retroperitoneal desmoplastic small round cell tumor. Methods One case of desmoplastic small round cell tumor was presented with review of the literature. Results A 11 years old child complained of abdominal pain and a 9.0 cm?6.0 cm?3.0 cm cystic tumor located in the left retroperitoneal space near the left kindey was resected.The connective tissue was significantly proliferated around the tumor cell nests. Immunohistochemical staining revealed CD 99 (+) and EWS/WT1 (+). After resection of the tumor,chemotherapy was performed with DDP?ifosfamide and VP16. No tumor recorrence has been noted 16 months after operation. Conclusions Desmoplastic small round cell tumor has specific clinicopathologic stigmata occuring usually in young males and the tumor was characterized by expression of MIC2 gene product CD 99 and the EWS/WT1 protein. Surgical resection with cheomotherapy is the treatment of choice.

Objective To investigate the changes of interleukin-21 (IL 21) and interleukin-21 receptor (IL 21R) expression level in Crohn's disease (CD) patients before and after accepted infliximab (IFX) treatment.Methods From June 2009 to July 2011,twenty-two CD patients met the research criteria were recruited at Tenth People's Hospital of Tongji University.Patients were treated with infliximab at weeks 0,2,6,and 16 healthy individuals were set as healthy control group at same time.Peripheral blood of healthy control group was taken at regular physical examination and blood of CD patients was taken before treatment and 10 weeks after treatment,intestinal mucosa biopsy samples were taken under colon endoscopy examination.The changes of Crohn's disease activity index (CDAI),erythrocyte sedimentation rate (ESR),C-reactive protein (CRP) in CD patients were observed.The change of IL-21R in peripheral blood CD4+ T lymphocytes was detected by flow cytometry.The change of IL-21 expression at mRNA level in intestinal mucosa was determined by realtime quantitative polymerase chain reaction (PCR).The data were analyzed by t test.Results Before treatment,the level of IL21R in peripheral blood CD4+ T lymphocytes of CD patients (12.25％±3.25％) and the expression of IL-21 at mRNA level in inflamed intestinal mucosa (1.38±0.32) were both significantly higher than those of healthy controls (4.25 ％ ± 1.41％,0.44±0.18),the differences were statistically significant (F=15.88,6.75 ; both P＜0.05).At 10th week,the level of IL-21R in peripheral blood CD4+ T lymphocytes of CD patients (8.12％ ± 2.05％) and the expression of IL-21 at mRNA level in intestinal mucosa (0.77 ± 0.24) were both significantly lower than those before treatment,the differences were statistically significant ( t=4.880,8.019; both P＜0.01).Before treatment,ESR,CRP and CDAI of CD patients was (46.8±11.4) mm/1 h,(52.4±11.5) mg/L and 319±74,which was (23.5±9.0) mm/1 h,(11.6±4.6) mg/L and 113±42 after treatment,the differences were statistically significant (t=9.485,16.458,11.100; all P＜0.05).Conclusion The IL-21 expression of active CD patients decreases after IFX treatment,which indicates that IL-21 may involve in IFX induced clinical remission of active CD.

Objective To discuss the indication and efficacy for treating hypospadias with ure-throplasty of Pedicled preputial double-faced island flap(PPDIF). Methods Records of 99 children (mean age, 5.1 years; age range, 4 months-12 years)with hypospadias who underwent PPDIF oper-ation were reviewed. Of them, 56 cases were penile type of hypospadias, 24 cases of penile root type, 13 cases of scrotal type and 6 cases of perineal type according to Barcat classification. Based on Duck-ett technique, the partial preputial cortical plate remained and was displaced to gastr-penis with ure-thra. , and finally the new urethra would be covered by the cortical plate. Results During follow-up of average 2 years, 8 patients(8. 1%) developed urethral fistula, 1 (1.0 %) developed mild meatal ste-nosis, and 1(1.0 %) developed flap necrosis. The success rate for total PPDIF procedure was 89.90 % (89/99). And the success rate of simple PPDIF for cases with penile and penile root hypospadias, and PPDIF+Duplay for cases with serotal and perineal hypospadias were respectively 91.25 % (73/80) and 84.21%(16/19). respectively. Conclusions PPDIF is good for treating most hypospadias. Furth-more it make penis better shape. In the severe cases, PPDIF+Duplay prededure is recommended and the effect was affirmative.

Objective:To investigate the correlation between the expression of angiogenic factors (MVD, IGF-1, and STAT3) in colorectal carcinoma and adenoma and the microvascular characteristics under narrow band imaging (NBI), in order to evaluate the fea-sibility of NBI in real-time observation of angiogenesis. Methods:Patients with pathologically confirmed colorectal polyps were re-cruited and examined by NBI. Vascular patterns were classified into typeⅠ(invisible or faintly visible), typeⅡ(clearly visible and regularly arranged in a round, oval honeycomb-like pattern), and typeⅢ(clearly visible and irregularly arranged in terms of size and caliber or irregularly winded). Immunohistochemical staining was performed to determine the expression of CD34, IGF-1, and STAT3. Histological results were compared with the vascular patterns under NBI. Results:The NBI endoscopy results of 64 sites (15 adenocar-cinomas, 29 adenomas, and 20 normal tissues) from 58 patients were introduced and examined in this study. Adenomas ranked the first (82.1%, 23/28) among the vascular patternⅡcases, whereas early adenocarcinomas dominated the vascular patternⅢcases (66.7%, 10/15). The expression levels of MVD-CD34 and IGF-1 in normal mucosa, adenomas, and adenocarcinomas were significantly differ-ent (P<0.0001 and P=0.0062, respectively). All the expression levels of MVD-CD34, IGF-1, and STAT3 in sites displaying vascular pat-ternsⅠ,Ⅱ, andⅢwere significantly different (P<0.0001, P=0.0010, and P=0.0055, respectively). Spearman correlation coefficients between the NBI vascular patterns and the MVD-CD34, IGF-1, and STAT3 expression levels were 0.67, 0.41, and 0.40, respectively. Conclusion:Vascular pattern analysis through an NBI system can be a promising tool to evaluate angiogenesis of colorectal lesions in real-time endoscopic observation.

Objective To investigate the expression of microRNA (miRNA)-10a in the intestinal mucosa,serum and peripheral blood mononuclear cell (PBMC) of patients with inflammatory bowel disease (IBD) and explore its role and relevance in the pathogenesis of the disease.Methods The intestinal or colonic mucosal biopsy specimens of nine active ulcerative colitis (UC) patients,11 active Crohn's disease (CD) patients and eight patients with negative colonoscopy result as control were collected.The sera of 12 active UC patients,13 active CD patients and nine healthy controls were collected.The PBMC of nine active UC patients,11 active CD patients and eight healthy controls were collected.The expression of miRNA-10a in the intestinal mucosa,sera and PBMC and the expression of IL-12/IL-23 p40 in the intestinal mucosa were detected by real-time polymerase chain reaction (PCR).Each 8 cases of active UC and CD patients were collected.The intestinal mucosa before infliximab (IFX) treatment and six weeks after three times of IFX treatment were collected.And at same time,the intestinal mucosa of 11 active UC patients and 10 active CD patients were collected and cultured for 18 hours stimulated with IFX in vitro and then the expression of miRNA-10a in the intestinal mucosa was tested.One-way analysis of variance was used for comparison in three samples.Paired t-test was used for two samples comparison.Spearman test was used for correlation analysis.Results Compared with healthy controls,the expression of miRNA-10a in the intestinal mucosa,serum and PBMC of UC and CD patients significantly decreased (F=38.45,30.46 and 14.74,all P＜0.05).There was no statistic significance between UC and CD groups.The expression of IL-12/IL-23 p40 in the intestinal mucosa of UC and CD patients significantly increased (F=32.90,P＜0.05).The expression of IL-12/IL-23 p40 was negatively correlated with the expression of miRNA-10a in the intestinal mucosa of CD patients.After three times of IFX treatment,the expression of miR-10a in the intestinal mucosa of IBD patients significantly increased (t=3.341,3.382,both P＜0.05).After stimulated with IFX in vitro,the expression of miRNA-10a in the intestinal mucosa significantly increased (t=3.095,7.193,both P＜0.05).Conclusions miRNA-10a was closely correlated with the inflammation of IBD patients and with the role of targeting IL-12/IL-23 p40.miRNA-10a might be a new target for the IBD treatment.

Objective To investigate the expression of complement regulatory proteins on placentas of pregnant C57BL/6 mice infected with Toxoplasma gondii in order to explore the molecular immunological mechanism for abnormal pregnancy induced by T. gondii infection. Methods Twenty-four pregnant C57BL/6 mice were randomly divided into two groups equally. The infection group was intraperitoneally injected with 200 of living T, gondii RH strain tachyzoites on the 8th day of gestation, and the normal group of mice was injected with physiological saline. All mice were killed on day 14 after gestation and placentas were collected. The expression levels of Crry, GPI-DAF and CD59a mRNA were analyzed by real-time quantitative PCR, and the positive rates of Crry and GPI-DAF were measured with flow cytometry. Results The died fetus rates of infected group and control were 80. 95% and 4. 41% , respectively. The infected group was significantly higher than of that the control group (P＜0.01). The expression levels of Crry, GPT-DAF and CD59a mRNA in the infected and control group were 0.786 ±0. 199, 0.594 ±0.096, 0.880 ±0. 179 and 0.550 ±0.077, 0.221 ±0.074, 0.591 ± 0.075 , respectively, and the difference of three kind of complement regulation proteins between two groups was all significant (P＜0.01). The positive percentages of Crry and GPI-DAF cells of infected and control group were (10. 03 ± 2. 11) % , (2.95 ±1.04)% and (3. 15 ± 1. 32) % , (0. 66 ±0. 26) % , respectively, and the difference of the two kind complement regulation proteins between two groups was also significant ( P ＜ 0. 01). Conclusion The expression level of mouse placental complement regulatory proteins was increased after infection with T. gondii, and then immunological microenvironment at the fetomaternal interface was destroyed. It may be one of important immunological mechanism for abnormal pregnancy induced by T. gondii infection.

Objective To create Mucin gene 1 (MUC1) antisense peptide nucleic acid (PNA),and to observe its effects on MKN-45 cell invasion and explore the mechanism. Methods The sequence of antisense PNA was designed according to MUC1 gene sequence and transfected into human gastric cancer cells (MKN-45) by liposome,and the empty vector group (randomized control group)and blank control group (negative control group) were involved. The expression of MUC1 was detected by real time quantitative PCR and the changes of E-cadherin expression were also observed.The effects on gastric cancer cell invasion were tested with transwell chamber assays.Results The expression of MUC1 gene was effectively suppressed by the 3 created antisense PNA,and their expression level (0.62±0.18,0.49±0.12 and 0.60±0.21) was significantly lower than that of negative control group (1.18 ± 0.03,P ＜ 0.01). There was no significant difference between radomized control group and negative control group (1.00±0.04,P=0.657).After MUC1 PNA transfected,the capability of gastric cancer cell invasion decreased significantly (P=0.005).And the expression of E-cadherin at mRNA and protein level was up-regulated.Conclusions There is negative correlation between MUC1 and E-cadherin expression in gastric cancer cell MKN-45.The capability of tumor cell invasion is significantly inhibited by suppressing MUC1 gene expression.