Objective To investigate the mechanism of the apoptosis of human primary gastric cancer cells implanted in nude mice induced by genistein. Methods Human primary gastric cancer cells were implanted into Balb/c nude mice to establish the cancer model, and the growth curve of implanted tumor was measured. Twenty-five nude mice with implanted gastric cancer cells were divided 5 groups. Genistein at different doses (0.5,1.0 and 1.5 mg/kg), normal saline and dimethyl sulfoxide were injected around the neoplasm. After treatment, transmission electron microscope and TUNEL staining were used to detect the apoptosis of implanted tumor cells. Immunohistochemical staining and RT-PCR were used to detect the expression of apoptosis-related genes bcl-2 and bax in implanted tumor. Results Genistein could significantly inhibit carcinoma growth when it was injected near the carcinoma. The apoptosis of tumor cells in implanted tumor induced by genistein was detected by transmission electron microscopy. The apoptosis index of the genistein treatment groups (28.7%?1.1%, 33.4%?1.4% and 37.1%?1.0%) was significantly higher than that of control groups(11.8%?0.4% and 11.7%?0.4%; P

ObjectiveThe purpose of the present study was to discuss the method to avoid spillage by means of adding chemotherapeutic drugs into sealed transfusion bottles.Methods90 penicillin sealed bottles of the same batch number were randomly divided into the test group and the control group,each with 45 bottles.Standard method according to Basic Nursing was used to add chemotherapeutic drugs into transfusion bottles in the control group.Drugs were added into transfusion bottles under negative pressure in the test group.The operating time was measured,the spillage volume of puncture site was calculated and microbial detection of syringe was observed in the two groups.ResultsThere was a significant difference in the spillage volume of puncture site between the two groups (P<0.01),but no difference was seen in the operating time and microbial detection of syringe (P>0.05),ConclusionsThe spillage volume of puncture site was reduced significantly by means of adding chemotherapeutic drugs into transfusion bottles under negative pressure.This can decrease chemotherapeutic professional exposing risks and drug wastage.

AIM: To investigate the apoptosis in primary gastric cancer cells induced by resveratrol, and the relation between this apoptosis and expression of bcl- 2 and bax. METHODS: In in vitro experiments, MTT assay was used to determine the cell gowth inhibitory rate. Transmission electron microscopy and TUNEL staining were used to quantitatively and qualitively detect the apoptosis of primary gastric cancer cells before and after the resveratrol treatment. Immunohistochemical staining and RT - PCR was used to detect the expression of apoptosis - regulated gene bcl - 2 and bax. RESULTS: Resveratrol inhibited the growth of primary gastric cancer cells in a dose - and time - dependent manner. Resveratrol induced primary gastric cancer cells to undergo apoptosis with typically apoptotic characteristics. TUNEL assay showed that after the treatment of primary gastric cancer cells with resveratrol for 24, 48, 72, 96 hours, the apoptotic indexs were 4.93% ± 0.19%, 16.74% ± 0.43%, 27.88% ±0.36%, 36.84% ± 1.07 % respectively. Immunohistochemical staining showed that after the treatment of primary gastric cancer cells with resveratrol for 24, 48, 72, 96 hours, the positive rates of Bcl - 2 proteins were 20.68% ± 0.49%, 10.84% ±0.33%, 6.80% ± 0.34%, 3.91% ± 0.15% and the positive rates of Bax proteins were 19.79% ± 0.98%, 30.74% ±0.85%, 40.14% ± 1.17%, 60.08% ± 1.64%. After exposed to resveratrol for 24 h, 48 h, 72 h and 96 h, the density of bcl- 2 mRNA decreased progressively with elongation of time and the density of bax mRNA increased progressively with elongation of time by RT- PCR. CONCLUSION: Resveratrol is able to induce the apoptosis in primary gastric cancer. This apoptosis may be mediated by down- regulation of Bcl- 2 and up- regulation of Bax.

Objective To explore the relationship between intrahepatic expression of interleukin-17 (IL-17) and liver fibrosis in patients with chronic hepatitis B virus infection. Methods IL-17 expressions in livers with different inflammation activity grades and hepatic fibrosis stages from patients with chronic hepatitis B virus carriers (n= 30), chronic hepatitis B (CHB, n = 55), liver cirrhosis (LC, n=20) were measured by immunohistochemistry. Serum IL-17 and liver fibrosis indices of haluronic acid (HA), laminin (LN), type Ⅲ procollagen (PC Ⅲ ) and type Ⅳ collagen ( Ⅳ C) were determined by enzyme linked immunosorbent assay (ELISA). The differences between groups were compared by Kruskal-Wallis test, and the Mann-Whitney test, and the correlation analysis was done by Spearman test. Results Intrahepatic IL-17 expression in LC group was significantly higher than CHB group (x2 =25. 3982, P=0. 004), and that in CHB group was higher than chronic hepatitis B virus carriers group (x2 = 11. 5056, P= 0. 001). The inflammation activity grade and hepatic fibrosis stage were both positively correlated with IL-17 expression (r= 0.718, 0. 693, respectively; both P＜0.01). IL-17 mainly located in portal area and the expression was positively correlated with serum levels of HA, LN, PCⅢ and ⅣC (r=0. 793, 0. 834, 0. 722, 0. 883, respectively; all P＜0.01).Conclusion Intrahepatic IL-17 expression is closely correlated with liver inflammation activity grade and hepatic fibrosis stage.

Objective To study the effect of neuroepithelial cell transforming gene 1 (Net1) on the cellular radiosensitivity and underlying mechanism.Methods Real-time quantitative PCR was used to measure the variations in Net1 expression level upon irradiation.Radiosensitivity was analyzed by colonyforming assay after Net1-siRNAs.Net1-associated proteins were identified by co-immunoprecipitation.Results The Net1 mRNA level in the cells was increased significantly (t =-10.52,P ＜ 0.05) after irradiation.Compared to the control group,siRNA-mediated silencing of Net1 enhanced cell radiosensitivity (t =15.31,11.65,P ＜0.05).Net1 was found to interact with Ku70,Ku80 and DNA-PKcs under either normal conditions or after irradiation.Conclusions Net1 could protect cells from irradiation by interaction with DNA repair proteins in non-homologous end joining pathway.

Objective To study the difference of expression of proteins in pancreatic cancer and adjacent fissue.Methods Proteomes of eight pairs of pancreatic ductal adenocarcinoma tissue samples and adjacent tissue samples were obtained by high resolution two-dimensional gel electrophoresis(2-DE).Comprehensive analyses of proteins were focused on total protein spots exhibiting statistical alternations between the two groups.Protein identification was done by peptide mass fingerprinting with tandem mass spectrometry(MS/MS).In addition,Western blotting and immunohistochemistry were performed to verify the expression of certain candidate protein.Results A total of 28 protein spot-features were found to be significantly increased and 17 significantly decreased in tumor tissues.Thirty of these protein spots were identified,which included enzymes,antioxidant proteins,signal transduction proteins,calcium-binding protein,structural proteins,chaperones and others.Western blotting and IHC further validated up-regulated expressions of one candidate protein(annexin II)in tumor tissues.Conclusions The analysis of proteomics with 2-DE on human tissue is a useful method for discovering valuable cancer marker candidates.These differential expressed proteins may serve as biomarkers for early detection and therapeutic targets to pancreatic cancer.

Objective To study the curative effect of XRCC2 gene silencing mediated by shRNA combined with radiation on human colonic trans?planted carcinoma in nude mice. Methods Colonic carcinoma T84 cells were transfered into BALB/c nude mice to establish a tumor xenograft mod?el in vivo. Mice were divided into three groups:control,shRNA?SC and shRNA?XRCC2 and exposed to X?ray radiation. The change of volume and weight of the xenografts were examined after receiving radiotherapy and the pathological analysis of tumor tissues were conducted. Results Tumor xenografts transfected with shRNA?XRCC2 in nude mice grew slowly. The xenograft volume in the shRNA?XRCC2 group was decreased significant?ly from day 12 to day 28 after radiotherapy compared with the control group(P<0.01). The xenograft weight in the shRNA?XRCC2 group was small?er than in the control group,with statistically significant difference(t=18.843,P<0.01). The inhibited rate of xenografts in the shRNA?XRCC2 group(56.25%),was markedly higher than that in the shRNA?SC group(4.69%). Pathological analysis of colonic transplanted carcinoma showed that nuclear atypia was not obvious,karyokinesis was decreased and small areas of necrosis were present in tumor xenografts treated with shRNA?XRCC2 transfection. Conclusion XRCC2 gene silencing combined with radiation has significant inhibition effect on colonic transplanted carcino?ma in nude mice.

Objective To investigate the clinical effect and the prospect of Infliximab in treatment of intravenous immunoglobulin (IVIG)-resistant Kawasaki disease (KD) patients.Methods Clinical features,inflammatory markers and coronary changes were observed in 2 cases of IVIG-resistant KD patients hospitalized in Peking University First Hospital,who were treated effectively by Infliximab.Relevant researches on the mechanism and progress of the Infliximab treatment for IVIG-resistant KD in the last 10 years were reviewed at the same time.Results Two KD patients hospitalized in Peking University First Hospital had been treated with 2 g/kg IVIG for 2 times and followed by methylprednisolone treatment.However,fever and other clinical manifestations occurred again after 2 days and 6 days when temperature returned normal.They both defervesced and all the symptoms were improved after 1 dose of Infliximab (5 mg/kg) by laboratory examinations.Four published literatures of the basic research and 9 retrospective or prospective clinical researches of Infliximab treatment of KD showed that Infliximab alleviated the inflammatory level in the KD patients significantly.Complete remission was up to 72.73％-92.11％.Those KD patients defervesced within 12 h,with dramatic improvement of symptoms and signs.Arthralgia also disappeared in the patients with arthritis.Only 1 case was complicated with hepatitis in the acute phase and cholecystitis in recovery time.A phase 3 randomised,double-blinded,placebo-controlled trial had been done to assess the addition of Infliximab to the standard therapy.Conclusions Infliximab is a feasible choice for IVIG-resistant KD patients.Efficacy and safety of Infliximab for KD treatment have been proved in the literature.However,Infliximab for KD treatment has not been indicated in the drug instruction,so the informed consent from the guardians and Ethics Committee is needed.

OBJECTIVETo analyze the differences between the oral microbiota of monozygotic and dizygotic twins by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE).

METHODSA total of 20 pairs of twin children were included in this study, in which 10 pairs were monozygotic (MZ) twins, and 10 pairs were dizygotic (DZ) twins. Of the 20 pairs, 10 pairs of twins had primary dentition, and 10 pairs had mixed dentition; 17 children had caries, and 23 children had no caries. Genomic DNA was extracted from saliva samples. The 16s rRNA was amplified and analyzed by PCR-DGGE. The PCR-DGGE band number and Shannon index were calculated.

RESULTSCluster analysis showed high similarity in the oral bacterial community seen in co-twins. However, no significant difference was seen between MZ and DZ twins. In the primary dentition, the PCR-DGGE band number and Shannon index of children with caries (11.00 +/- 1.56, 1.05 +/- 0.36) were lower than those of children without caries (14.00 +/- 2.74, 1.44 +/- 0.37) (P < 0.05). In mixed dentition, the PCR-DGGE band number and Shannon index of children with caries (11.88 +/- 4.05, 1.18 +/- 0.36) were lower than those of children without caries (14.31 +/- 5.71, 1.28 +/- 0.47), but the differences were not statistically significant (P > 0.05).

CONCLUSIONEnvironmental factors may have a stronger effect on the constitution of oral microbiota in children compared with genetic factors. Children without caries may have a richer microbial diversity compared with children with caries.

Bacteria ; Child ; Denaturing Gradient Gel Electrophoresis ; Dental Caries ; Female ; Humans ; Male ; Microbiota ; Mouth ; microbiology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; Saliva ; Twins, Monozygotic

Objective:To study the inhibitory effect of retinoblastoma 94(Rb94) gene combined with radiotherapy ionizing radiation on the growth of esophageal carcinoma cells of tumor-bearing nude mice, and to clarify the synergistic effect of Rb94 gene and radiotherapy in inhibiting the growth of tumor cells.Methods:The models of tumor-bearing BALB/c-nu nude mice were built by inoculating the K150 cells.The model mice were divided into five groups:blank control(no any treatment), Ad-LacZ(control adenovirus including LacZ gene but not Rb94 gene, Ad-LacZ was transfered into tumor xenograft on 0, 3, 7 d separately), Ad-Rb94(tumor xenograft was transfected with Ad-Rb94 on 0, 3, 7 d separately), radiation (tumor xenograft was irradiated with 4 Gy γ-radiation on 1, 4, 8 d separately) and Ad-Rb94 combined with radiation(combination group, tumor xenograft was irradiated with 4 Gy γ-radiation after transfected with Ad-Rb94) groups.The volumes and the weights of esophageal carcinoma and the inhibitory rates of tumor growth of the mice in various groups were detected.The expression levels of ABL and JNK kinase in tumor tissue of the mice in various groups were measured,and the pathological changes of tumor tissue were investigated.Results:The speeds of tumor growth of the nude mice in Ad-RB94, radiation, and combination groups were slower than that in control group.The volume of esophageal carcinoma in combination group at day 15 after treatment was markedly smaller than those in Ad-RB94 and radiation groups,and there were significant differences compared with control group and Ad-LacZ group (F=26.7,23.8;P<0.01).The tumor weight of the nude mice in combination group was the lightest at the end of treatment;the inhibitory rate of tumor growth in combination group reached 81.16% and was significantly higher than those in Ad-Rb94 group(57.84%)and radiation group(38.20%)(P<0.01).The expression levels of ALB and JNK kinase in tumor tissue of the mice in combination group was markedly higher than those in control group(P<0.01).Compared with other groups, the tumor cells in combination group had fewer karyokinesis and lower level of nuclei hyperchromasia.Conclusion:Rb94 gene combined with radiotherapy shows synergistic effect in inhibiting the growth of tumor of tumor-bearing nude mice.