Child, Preschool ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; virology ; Male

Objective To study the surgical techniques and effects of laparoscopy-assisted sigmoid colon vaginoplasty.MethodsClinical data of 27 patients,who underwent laparoscopy-assisted sigmoid colon vaginoplasty at Beijing Anzhen Hospital between June 2006 and June 2007,were retrospectively analyzed.A 15-cm segment of pedicled sigmoid colon was isolated using an ultrasound knife.The distal end of the segment was pulled into the vaginal space in the cul-de-sac of Douglas under a laparoscopic vision as the neovagina.The continuity of the intestinal tract(end-to-end bowel anastomosis) was restored using a circular mechanical suture through the rectum.ResultsThe surgery was successfully completed in all the cases,no intra-operative complication occurred.The mean blood loss and operation time was 82 ml(50-180 ml) and 168 min(120-246 ml) respectively.One patient developed incomplete intestinal obstruction 16 days after the operation,and was cured by conservative treatment.Follow-up was available in 21 patients for 14-20 months.Five patients had no sexual partner during the follow-up,while the other 16 patients were satisfied with their sexual lives after the surgeries.ConclusionLaparoscopic vaginal reconstruction using a sigmoid colon segment is satisfying for cosmetic,functional,and anatomic results.

BACKGROUND:Tumor necrosis factor-α(TNF-α), a main cytokine inducing chondrocyte apoptosis of osteoarthritis, plays a regulatory role in the process of osteoarthritis. OBJECTIVE:To compare the rat models of chondrocyte apoptosis induced by different mass concentrations of TNF-α, thus providing theoretical basis for further study on the regulation of drugs on chondrocyte apoptosis. METHODS:Chondrocytes were isolated from the knee cartilage of 4-week-old Sprague-Dawley rats of clean grade by mechanical l col agenase digestion and were then incubated with different mass concentrations of TNF-αto induce apoptosis. The morphology of chondrocytes was observed under inverted phase contrast microscope, cel s were identified using immunohistochemical staining of type II col agen, as wel as the cel viability and apoptosis were detected by MTT and DAPI, respectively. RESULTS AND CONCLUSION:(1) In vitro, the cytoplasm of chondrocytes was stained brown-yel ow by using immunohistochemical staining of type II col agen. (2) At 48 hours, the apoptosis rate of chondrocytes induced by 10, 20 and 30μg/L TNF-αwas significantly higher than that of the 0μg/L TNF-α(P<0.01), and the apoptosis rate of chondrocytes induced by 40μg/L TNF-αwas significantly higher than that of the 10μg/L TNF-α(P<0.01). (3) The viability of chondrocytes induced by 10, 20 and 40μg/L TNF-αwas significantly lower than that of the 0μg/L TNF-α(P<0.01). In detail, the viability of chondrocytes induced by 20μg/L TNF-αwas lower than that of the 10μg/L TNF-α(P<0.05);the viability of chondrocytes induced by 40μg/L TNF-αwas significantly lower than that of the 10 and 20μg/L TNF-α(P<0.01, P<0.05). (4) These results suggest that 20μg/L TNF-αcan successful y induce the chondrocyte apoptosis model.

Polysaccharides with multiple biological activities are usually considered as one of the major bioactive compounds in Chinese medicines (CMs). At present, the development of drug and functional foods related to polysaccharides have attracted a great deal of attention due to their great potential effects and diverse action mechanisms. However, quality control of polysaccharides is the bottleneck and a challenge due to their complexity and chemical diversity. Actually, the bioactivities of polysaccharides are closely related to their molecular structures. In order to ensure their safety and efficacy, the development of novel approaches based on the molecular structures for the improvement of quality control of polysaccharides is significantly important. Therefore, in this article, the relationship between biological activities and chemical structures, as well as the action mechanisms of polysaccharides from CMs were summarized first. Furthermore, saccharide mapping, a novel strategy for quality control of bioactive polysaccharides from CMs, was introduced and the application and perspectives were also discussed.
Carbohydrate Sequence ; Drugs, Chinese Herbal ; chemistry ; Molecular Sequence Data ; Polysaccharides ; chemistry ; Quality Control

Cytokine-Induced Killer Cells ; immunology ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Fetal Blood ; immunology ; HL-60 Cells ; Humans ; K562 Cells ; Killer Cells, Natural ; immunology

Objective To identify the effect of arsenic trioxide (As2O3) on the differentiation and apoptosis of different types of neuroblastoma(NB) cell lines.Methods The cell lines [SK-N-SH,SK-N-BE2,SH-SY5 Y] were induced with different concentrations (0 μmol/L,1 μmol/L,3 μmol/L,5 μ mol/L and 7 μ mol/L) of arsenic trioxide for 24 h,48 h,72 h under the same conditions.The expression of MYCN gene was examined by fluorescence in situ hybridization assay in SK-N-BE2,cell proliferation,cell cycle and cell apoptosis were detected with cell counting kit-8 (CCK-8) assay and flow cytometry.Results 5 μmol/L of As2O3 inhibited the expression of MYCN gene in SK-N-BE2;CCK-8 assay indicated that As2O3 inhibited the proliferation of NB cell in a dose-and time-dependent manner,the cell proliferation was significantly suppressed compared with the low concentration (1 μ mol/L) after treated with As2O3 by 1 μmol/L,3 μmol/L,5 μmol/L and 7 μmol/L in 24 h,48 h and 72 h,SH-SY5Y:24 h(chisq =9.666 7,P ＜ 0.05),48 h (chisq =9.666 7,P ＜ O.05),72 h (chisq =9.512 8,P ＜ 0.05);SK-N-SH,24 h (chisq =10.38,P＜0.054 6),48 h(chisq=8.641 0,P＜0.05),72 h(chisq=9.461 5,P＜0.05);SK-N-BE2:24 h (chisq =8.435 9,P ＜0.05),48 h(chisq =8.641 O,P ＜0.05),72 h(chisq =9.545 5,P ＜0.05);compared with the control group,the As2O3-treated cells showed increased apoptosis percentage,with the percentage of 1.6％ (0 μmol/L),3.8％ (1 μmol/L),6.1％ (3 μmol/L),10.4％ (5 μmol/L),40.2％ (7 μ mol/L);the cell cycle was arrested at G2/M phase,which prevented cell division.Conclusions (1) As2O3 play an important role on the NB cells proliferation,apoptosis which were dose-and time-dependent manner.(2)As2O3 can inhibit the expression of MYCN gene.(3)As2O3 also could block NB cell cycle at S and G2/M,and inhibit the cell nucleus replication and the As2O3 had different induced effect between different types of NB cell.

Objective To investigate the effects of 13-cis retinoic acid (13-cis RA) in inducing differentiation of 3 types of human neuroblastoma (NB) cells in vitro.Methods The status of MYCN gene amplification of cultured SH-SY5Y,SK-N-SH and SK-N-BE2 cells was detected by fluorescence in situ hybridization.After treatment with different concentrations of 13-cis RA,morphological changes were observed by phase-contrast microscope,and neuron-specific enolase (NSE) concentrations were determined by enzyme linked immunosorbent assay.The cell viability was measured through cell counting kit-8 assay,and the cell apoptosis was assayed with flow cytometry (FCM).Results The morphological changes in differentiation were observed in all 3 types of NB cells after 13-cis RA treatment.MYCN amplification was detected in SK-N-BE2 cells even after 13-cis RA treatment,while the other 2 cell lines were amplification-null.After different concentrations of 13-cis RA treatment,NSE concentration increased with prolonged time,especially for SK-N-BE2 cell(F =27.00,P ＜ 0.000 1).13-cis RA stimulated cell proliferation within 48 hours of treatment,and then inhibited cell growth.FCM indicated that the degree of apoptosis in SH-SY5Y cell was significantly higher after 13-cis RA treatment of 10 μmol/L concentration for continuous 96 h and 120 h as compared to the control group (F =16.21,P =0.011;F =16.04,P =0.016).Cell apoptosis of SK-N-SH cell after 13-cis RA treatment of 1 μ mol/L and 10 μ mol/L concentration for 48 h,were significantly higher than those of the control group (F =15.05,P =0.012;F =31.18,P =0.005);while SK-N-BE2 cell with different concentrations of 13-cis RA(1 μmol/L,5 μmol/L,10 μ mol/L) for 120 h were significantly higher than those of the control group(F =9.05,P =0.030;F =11.38,P =0.028;F =7.88,P =0.041).Conclusions The present study showed that 13-cis RA could induce differentiation of human NB ceils in vitro.It induces cell proliferation within 48 hours of 13-cis RA,and thereafter suppresses cell growth.No improvement was found in MYCN amplification cells with the detection of DNA level after 13-cis RA treatment,which suggests that combined treatment is possibly needed.

Objective To investigate the detection of early cervical cancer patient's peripheral blood CK19 mRNA and its clinical significance. Methods 30 early (ⅠA2~ⅡA) cervical cancer patients from the department of gynecological tumors in the third affiliated hospital of Kunming Medical University were selected as experimental group, 15 patients with uterine myoma and 15 healthy volunteers as negative control group,15 advanced (ⅡB~ⅢB) cervical cancer patients as positive control group. Peripheral blood samples before receiving radical resection of experimental group,and the same samples before treatment of control groups were collected. The change of relative expression level and positive rate of peripheral blood CK19 was analyzed using qRT-PCR technique, and its correlation with clinicopathological parameters and prognosis was further analyzed. Results There was significant difference in relative expression level of CK19 mRNA in the peripheral blood between experimental group and control groups ( <0.001) . And the positive rate of CK19 in the peripheral blood from 30 patients with early cervical cancer before operation in the experimental group was 30% (9/30) . Furthermore, CK19 mRNA positive rate before surgery was correlated with lymph node metastasis, muscularis invasion and pathologic risk factors for postoperative ( <0.05) . There were no significant correlation between CK19 mRNA positive rate and other clinical and pathological features such as age,clinical stage (FIGO),tumor cell differentiation,tumor size,pathological type and etc ( > 0.05) . Conclusions CK-19 mRNA is a specific and suitable molecular marker for the detection of circulating tumor cells in early cervical cancer. Detection of peripheral blood CK19 mRNA expression before receiving radical hysterectomy in patients with early cervical cancer can also make up for the disadvantage of other methods such as the imaging, tumor marker detection. Most important of all, it can provide the basis for choosing a proper adjuvant therapy post-operation and estimating the relapse and prognosis.

Background Mutant C57BL/6 mouse with corneal opacity (B6-Co) appears eye open at birth (EOB) phenotype,which is a good animal model in the study of developmental mechanism of eyelid.Investigating the relationship between serum response factor (SRF) and EOB phenotype can provide theoretical support for the research on the mechanism of innate defects in eyelid development in humans.Objective This study was to assess the dynamic expressions of SRF in eyelid of embryonic B6-Co mouse.Methods Total RNA was extracted from B6 and B6-Co mice eyelid tissue at embryonic day 16.5 (E16.5 d),E17.5 d and E18.5 d.The relative expression levels of SRF mRNA and protein in the eyelid tissue of B6 and B6-Co embryonic mice were assayed by real-time quantitative PCR and Western blot,respectively.In situ expressions of SRF protein in eyelid of B6-Co mice and B6 mice were detected using immunofluorescence technique.The use and care of the animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals of Nantong University.Results The relative expression levels ofSRF mRNA in the eyelids were 0.41±0.06 and 0.24±0.17 in E16.5 d and E17.5 d of B6-Co mice,showing a significant decline in comparison with 1.03 ±0.17 and 1.01 ±0.09 in the B6 mice (P =0.025,0.017).The expression levels of SRF protein in the eyelids of E16.5 d and E17.5 d B6-Co mice were 0.08±0.01 and 0.08± 0.01,which were significantly lower than 0.12 ±0.03 and 0.13 ± 0.02 of B6 mice (P =0.036,0.024).However,there were no significant differences in the expression levels of SRF mRNA and protein in E18.5 d between the B6-Co mice and B6 mice (P =0.387,0.774).Immunofluorescence assay displayed that SRF was expressed in the keratinocytes of eyelids in both mice,but the fluorescence intensity was weaker in the B6-Co mice.Conclusions SRF probably interrupts the developing process of eyelid in early embryo of B6-Co mice.

Objective Study the ideal drug to treat age depen de nt epileptic ncephalopathy (ADEE) in earlier period. Methods fi fty-four patients with ADEE were studyed.21 cases used with single topiramate(T PM) as initial treatment and 27 cases used with TPM and antiepileptic drug(AEDs ) .Results Seizure was fully controlled in 11 patients with TPM m onotherapy and in 4 patients with TPM as adjunction.The differences was signif icant(P