Animals ; Dependovirus ; genetics ; immunology ; Gene Transfer Techniques ; instrumentation ; Genetic Therapy ; instrumentation ; Genetic Vectors ; genetics ; immunology ; Humans

By molecular cloning technique, the expressing plasmids pDOR-SV40-Bcl-2 cDNA were successfully constructed. The reconstructed plasmids with lipofectamine were transduced into gastric cancer cell line SGC7901 and then the positive clones which contained the reconstructed plasmids were choosed by G418. Circa 150 positive clones were choosed from 2 x 10_5 gene transduced cells, which suggested that the transducing efficiency was more than 1%o; 2 positive clones were expanded and passage, then 1 drug-resistance cell strain (SGC7901 anBcl-2 cells) was obtained. The bloting results suggested, Bcl-2 cDNA were expressed in both gene transduced and not transduced cell strains, but the expressing level of mRNA and protein in gene transduced cell strain was very low than that in gene not transduced cell strain were positive by the means of Southern blot, Northern blot and Western blot. This results showed that normally in gastric cancer cells the expressing level of Bcl-2 gene was very high, and the cDNA fragment of antisense Bcl-2 were successfully transduced into gastric cancer cells, and in gene transduced cell strains the expressing of Bcl-2 gene was effi-cientlv blocked.

0.05).3.Thirty-three point three percent of animals from parental strain group were found fibrillations potentials and the po-sitive sharp waves in gastrocnemius electromyogram,no obvious abnormal waves were found in animals from both waaF mutant and control group.Conclusions The ganglioside-like epitope in LOS of CJ is critical antigen in inducing GM1-IgG antibody and in inducing conduction block of peripheral nerve,therefore,provide a support for the molecular mimicry theory as a pathogenesis in the axonal GBS following CJ infection.

Objective To study the killing effect of focused ultrasound activated-hematoporphyrin on H-22 and its optimum exposure time.Methods The distribution of hematoporphyrin in H-22 cells was measured by a fluorescence photometer,the uhrastructure changes were evaluated at different time with a scanning electron micro- scope after treatment with focused ultrasound at the frequency of 1.43 MHz and intensity of 1.0 W/cm~2, 2.0 W/cm~2,3.0 W/cm~2,respectively,in combination with HpD.Results The concentration of HpD in the H-22 cells reached its peak after being added to H-22 tumor cells for 45 minutes,which will produce the hest anti-tumor effect when activated by ultrasound.Morphological observation showed that HpD alone had a slight influence on H-22 cells;ULtrasound alone showed an anti-tumor effect on tumor cells,which was dosage-dependent.Ultrasound dia- thermy in combination with HpD has more effective in terms of its antitumor effect when compared with uhrasound dia- thermy alone.Conclusion The killing effect on H-22 tumor cells of sonodynamic treatment was dependent on the intensity of uhrasound and the content of HpD in the cells,as well as on the time of action of both uhrasound and HpD.

Gene Library ; Humans ; Nucleic Acid Hybridization ; methods ; Pulmonary Fibrosis ; diagnosis

BACKGROUND: Embryonic stem cells (ESCs), the seed cells of all mature cells in vivo, are useful tools for nerve transplantation and developmental gene function research. Notch1 signaling pathway is the key pathway to control the ordered neural development and differentiation of many kinds of neural cells, however, there is no report on the dynamic expression of Notch1 signal during the ESC differentiation to date. OBJECTIVE: To investigate the expression of Notch1 protein, transmembrane signal transduction molecule, during directional differentiation of embryonic stem cells into neural cells. DESIGN, TIME AND SETTING: Cell research was carried out between October 2003 and October 2004 at Zhongshan Ophthalmic Center, SUN Yat-sen University, Guangzhou, Guangdong Province, China. MATERIALS: BALB/C mouse embryonic stem cell line Ⅵ (passage 11)was obtained from experimental animal center of SUN Yat-sen University, provided by professor Huang Bing. ESC culture medium was high-glucose DMEM medium with 20% bovine serum and 106 IU/L mouse leukemia inhibitory factor. Induced differentiation medium was high-glucose DMEM medium with 20% fetal bovine.serum and 5×107 mol/L retinoid acid(RA). METHODS: Passage 11 ESCs were resuscitated and incubated by ESC culture medium in incubator at 37℃ with 5% CO2. Passage 11 ESCs were subcultured after 2 or 3 days and RA was added into medium to induce differentiation. Three time points for observation were established: induced for 1, 5 and 9 days. MAIN OUTCOME MEASURES: Morphological changes were observed under inverted phase contrast microscope, MAP-2 antigen expressed in differentiated cells was detected by immunofluorescence method. Immunocytochemistry, Western Blot, flow cytometry assay were used to investigate the Notch1 protein expression. RESULTS: ESCs presented clone-like growth. After induced by RA for 9 days, single neural network was achieved around most of the cell clusters. With the prolongation of induction, MAP-2 positive neural cells increased gradually. Almost all ESC clones expressed Notch1 protein strongly or positively, but Notehl protein expression decreased gradually after induced differentiation (P < 0.01). CONCLUSION: Notch1 signal shuts off progressively during induction of ESCs into neural cells, which suggests Notch1 may play an important role in the differentiation of ESCs into neural cells.

Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.

Bronchoalveolar Lavage Fluid ; Humans ; Interleukin-1beta ; metabolism ; Macrophages, Alveolar ; cytology ; drug effects ; metabolism ; Silicon Dioxide ; adverse effects ; Silicosis ; metabolism

Objective To investigate the influence of 1oganin on proliferation and differentiation of rat preadipocytes. Methods Rat preadipocytes were cultured. The proliferation of rat preadipoeytes was detected by MTT method. The expression of GPDH during the preadipoeytes differentiation was determined by using enzyme tissue chemical way, and the accumulation of cellular lipid was determined by Oil Red O staining. Results Loganin at the concentrations of 8, 16, 32 ? mol/L stimulated rat preadipocyte proliferation, inhibited GPDH increase and lipid accumulation during differentiation in a concentration-dependent manner. Conclusion Loganin can stimulate rat preadipocyte proliferation, inhibite rat preadipocyte differentiation.

Objective To develop and verify an influenza A virus antigen-detecting kit which can detect all the subtypes of influenza A virus. Methods Double-antibodies sandwich enzyme linked immunosorbent assay (ELISA) was utilized for developing the influenza A virus antigen-detecting kit. The sensitivity, specificity, accuracy and stability of the kit were evaluated by the clinical samples. Results The lower detection limit of the kit for the N protein was 7. 63 ng/mL, which was 256 times lower than that of the hemagglutination and 16 times lower than that of immune colloidal gold technique. The kit didn't show any cross-reaction with the influenza B virus, respiratory syncytial virus, respiratory adenovirus, para-influenza virus type Ⅰ and Ⅲ, mycoplasma pneumomiae, avian newcastle disease virus, avian infectious bursal disease virus or avian infectious bronchitis virus. The specificity was 100%. Both the intra batch variaton coefficient (CV) value and inter batch CV value were less than 15%, which met the national standard for ELISA kits. The results proved that the kit could keep stable at 4 ℃ for more than 1 year and at 37 ℃ for more than 7 days. The kit could identify H1N1, H3N2, H5N1 and H9N2 influenza A viruses. The clinical research data of human influenza virus showed the consistency rate between the kit and regular cell culture method was 93. 44% for the positive samples and 99. 31% for the negative samples. The clinical research data of avian influenzavirus showed the consistency rate between the kit and regular cell culture method was 95. 45% for positive samples and 98. 09% for negative samples. Conclusion The influenza A virus antigen-detecting ELISA kit can be used for the epidemiological survey of the infection of human influenza A virus or avian influenza virus with high sensitivity and specificity.