Through analysis on the conflicts in the First-Aid,the article holds that the causes include not only the insufficiency of the doctor's responsibility,emergence consciousness and service consciousness,but also the deficits of medical service and unreasonable medical cost.Furthermore,the author points out that the underlying cause is the irrational architectonic ethics,puts forward the countermeasures,which could prevent the medical conflicts in First-Aid.

Objective To explore the effects of X-ray irradiation combined with RNAi against signal transducer and activator of transcription 3(STAT3)on the radiosensitivity of human esophageal carcinoma cells.Methods Human esophageal carcinoma cells of the line Eca-109 were euhured.Three pairs of DNA template aiming at the base sequences of the coding regions 2037-2055,1243-1261,and 455-473 of the STAT3 mRNA were synthesized(siRNAI,siRNA2,and siRNA3),and a negative sequence was synthesized to be used as control.STAT3-siRNA positive recombinant plasmids(pRNAT-U6.1-siRNAI,pRNAT-U6.1-siRNA2, and pRNAT-U6.1-siRNA3), and a STAT3-siRNA negative recombinant plasmid (pRNAT-U6.1-negative)were thus constructed and then transfected into the cultured Eca-109 cells,which were divided into transfection reagent control group,pRNAT-U6.1-siRNAl-3 transfection groups,and pRNAT-U6.1-negative centrel group.The positive eell clones were screened.RT-PCR and Westem blotting were used to detect the STAT3 mRNA and protein expression.The transfected Eca-109 cells were exposed to 0,2,4,6,and 8 Gy of X-rays,respectively,and the survival fraction of the cells was analyzed by clone formation assay.Flow cytometry was applied to analyze the cycle arrest and cell apoptosis 4 Gy post-irradiation.Results Agarose gel electrophoresis confirmed the successful construction of the plasmid pRNAT-U6.1-siRNA.RT-PCR and Western blotting demonstrated that the mRNA and protein expression levels of STAT3 transfected with sTAT3-siRNA3 were both significanfly lower than those of the control groups.At 2-8 Gy, the survival fractions of the siRNA3 group were aU significantly lowered than those of the control group(t＝-0.228--0.051,P<0.05).Flow cytometry showed that the percentage of the cell cycle G0/G1 phase and the apoptosis rate of the siRNA3 group were both significantly higher than those of the control groups at 4 Gy post-irradiation(t＝-13.137-16.350,P<0.01).Conclusions X-ray irradiation combined with RNAi against sTAT3 could inhibit the proliferation of the human esophageal carcinoma cells,induce cell cycle arrest and apoptosis,improve the radiosensitivity in Eta-109 cells.

Objective: To study the cell proliferation, cell cycle and apoptosis of esophageal carcinoma Eca-109 cells treated with RNA interference technique to silence signal transducers and activators of transcription 3 (STAT3) gene. Methods: Three pairs of DNA template coding siRNA specific for human STAT3 gene mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pRNAT-U6.1/neo plasmid to construct STAT3-siRNA expression vector which was then transfected into Eca-109 cells. The expression of STAT3 mRNA and protein in cancer cells was detected by RT-PCR and Western blot, respectively. The cell proliferation, cell cycle distribution and apoptosis were examined by MTT and flow cytometry. Results: STAT3-siRNA expression vector was successfully constructed and identified by sequencing. The results of RT-PCR and Western blot demonstrated that STAT3 expression in Eca-109 cells transfected with STAT3-siRNA expression vector was significantly higher than that in the control group (P＜ 0.01). MTT showed that after transfection of the siRNA vector into Eca-109 cells, cell proliferation was obviously reduced and the cell growth inhibition ratio in the siRNA3 group was 35.68%, significantly higher than that in the control group (P＜0.01). Flow cytometry results suggested that cell cycle arrest and more apoptosis were observed in the siRNA3 group. Cell cycle was arrested at G_0/G_1 phase, and the rate of apoptosis was 13.26%, much higher than that in the control group (P＜0.01). Conclusion: Silencing STAT3 gene by RNA interference technique can effectively inhibit STAT3 expression, suppress the proliferation of Eca-109 cells, induce cell cycle arrest at G_0/G_1 phase, and promote apoptosis.

Background and purpose:Several reports demonstrated that the expression of STAT3 has been found to be an oncogene in solid tumors such as head and neck squamous cell carcinoma,esophageal carcinoma and prostate carcinoma.This study was done to explore the effects of X-ray irradiation combined with RNAi against STAT3 on proliferation and apoptosis of human esophageal carcinoma cell line Eca-109.Methods:Three pairs of DNA template coding siRNA were synthesized against STAT3 to reconstruct pRNAT-U6.1-siRNA-STAT3,which was transfected into Eca-109 cells,the positive cell clones were screened with G418.Inhibitory effect of STAT3 mRNA and protein in Eca-109 cells was detected by RT-PCR and Western blot,respectively.The transfected cells were exposed to 0,2,4,6 and 8 Gy X-ray respectively;the survival fraction of Eca-109 cells was analyzed by clone formation assay,and flow cytometry was applied to analyze cell apoptosis at the dose of 4 Gy.Results:pRNAT-U6.1-siRNA-STAT3 was reconstructed and identifi ed as correct by sequencing.RT-PCR and Western blot analyses demonstrated that STAT3-siRNA could obviously reduce the expression of STAT3 mRNA and protein in Eca-109 cells.Clone formation assay and flow cytometry results showed that irradiation at different doses combined with STAT3-siRNA could inhibit the proliferation of Eca-109 cells,irradiation with 4 Gy X-ray could induce apoptosis.Conclusion:X-ray irradiation combined with RNAi against STAT3 could inhibit the proliferation of Eca-109 cells and induce apoptosis.

Objective To study the relationship between serum HBV DNA level and apoptosis of peripheral blood mononuclear cells (PBMC) , and the relationship between serum HBV DNA level and the activity of caspase-8 in the patients with chronic hepatitis B (CHB). Method 30 CHB patients were selected as experimental group, and it was divided into three subgroups according to the serum HBV DNA level, subgroup A (high serum HBVDNA), subgroup B (medium serum HBVDNA), and subgroup C (low serum HBVDNA). 10 healthy adults were random selected as control group. PBMC were isolated from two groups by separating medium of lymphocytic cell and culturing it with phytohemagglutinin (PHA) in vitro for 72 hours. The PBMC was stained with PI and the apoptosis was assayed with flow cytometry. At the same time, the aetivity of caspase-8 of PBMC was assayed by color matching. Results The apoptosis rate of PBMC of experimental group ( 26. 88 ± 7.37 ) % were higher than that of the control group ( 14. 95 ±2. 53)% ( P <0. 01 ). In the experimental group, the apoptosis rate of PBMC of subgroups A, B and C showed decreasing order (34. 75 ± 4. 59)%, (25.63 ± 3.55 )%, ( 18. 91 ± 3. 81 )%. The activity of caspase-8 of experimental group 2. 99 ±0. 82 were higher than that of the control group 1.43 ±0. 91 ( P <0. 01 ). The activity of caspase-8 of subgroup A, B and C showed the same decreasing order: 3. 87 ±0. 35,2. 95 ± 0. 36, 1.95 ± 0. 29. There was a positive correlation between the apoptosis level of PBMC and the activity of caspase-8 in experimen tal group ( r = 0. 610, P < 0. 01 ). Conclusion AICD of PBMC was found in patients with CHB. The activity of caspase-8 increased in that process, and it may participate in the transduction of apoptosis signal. Serum HBV DNA level was related with the apoptosis rate of PBMC and the activity of caspase-8, and it may be one of the reasons of apoptosis in PBMC.

Objective To probe into the content of DNA Topo Ⅱ in osteosarcoma after chemotherapy.Methods 30 patients with osteosarcoma received two courses of chemotherapy treatment before the surgical resection of the tumor tissue.Then immunohistochemistry was used to detect the content of Topo Ⅱ in tissues and detected its relationship in pathology.Results There were 8 out of 30 cases in which Topo Ⅱ was presented positive in osteosarcoma (26.7 ％).The protein content of Topo Ⅱ was unrelated to the patient' s age,gender,degree of tumor malignancy,tumor location and translocation or Enneking staging (P ＞ 0.05),but related to patients survival rate (P ＜ 0.05).Conclusion Patients with lower expression of Topo Ⅱ are more likely to have poor prognosis.

[ ABSTRACT] AIM:To investigate the effects of microRNA145 ( miRNA145 ) on the viability, apoptosis, inva-sion and metastasis of hepatoma HepG2 cells.METHODS: HepG2 cells were randomly allocated into 3 groups: blank control group, empty mimic transfected group and miRNA145 mimic transfected group.Under the induction of Lipofectami-neTM 2000, the recombinant was transfected into HepG2 cells.After transfection, the expression level of miRNA145 was detected by real-time PCR.The protein level of N-cadherin and the mRNA expression levels of miRNA145 and N-cadherin were detected by Western blot and real-time PCR.The cell viability was detected by MTS assay.The cell cycle and apopto-sis were analyzed by flow cytometry.Invasion and metastasis were detected by Transwell assay.RESULTS:Compared with negative control, miRNA145 expression was up-regulated significantly, while the expression of N-cadherin was down-regu-lated significantly.Meanwhile, the cell viability, cell cycle, apoptosis, invasion and metastasis of hepatoma HepG2 cells were all significantly inhibited (P<0.05).CONCLUSION:miRNA145 dramatically inhibits viability, apoptosis, inva-sion and metastasis of hepatoma cells.

Objective To analyze the genotype of Japaneso encephalitis virus (JEV) strains isola-ted in 2004 from mosquitoes collected in Bazhong city, Sichuan province of China, and the characters of amino acid in the PrM and E gene. Methods The isolated virus strains from mosquitoes were identified by biological, serological and molecular biology. PrM and E segments of the isolated JEV were amplified by RT-PCR, the PCR products were purified and sequenced. Multiple alignment, phylogenetic and amino acid (AA) analysis were carried out by Clustal X (1.8) , MEGA4 and GENEDOC (3.2) . Results The total of 4688 mosquitoes were collected including Armigeres and Culex. Six isolates were identified be-longing to genotype 1 JEV. The comparison between new genotype 1 JEV strains and live attenuated vaccine strain SA14-14-2 in PrM and E gene showed that total 3 sites amino acid differences in PrM gene and 14 sites in E gene, respectively. Three sites (PrM2, 64 and 65 ) in PrM protein and four sites (E129, 222,327 and 366) in E protein were only belonging to genotype 1 JEV. Conclusion The new isolated JEV strains in Sichuan province belong to genotype 1. It suggests that the vaccine strain SA14-14-2 currently used for preventing Japanese encephalitis is able to protect people against JEV, although in the segments of it had some amino acid differences between vaccine strain and the epidemic genotype 1 JEV strains in PrM and E gene.

Objective To observe the effects of ziprasidone and risperidone on patients with schizophrenia and their influence on bloodglucoseandlipids.Methods 96patientswithschizophrenicenrolledinthestudywererandomlydividedintotwogroups,zi‐prasidone and risperidone group ,and both were treated for 8 weeks .Their blood glucose ,blood lipid of base line and at the end of the 4th ,8th week were determined respectively .Results The positive and negative symptoms scores of the two groups by using Positive and Negative Syndrome Scale(PANSS) before and after treatment were not statistically different(P> 0 .05) .Compared with the baseline scores ,scores at the end of 4th and 8th week in both ziprasidone and risperidone groups significantly decreased(P<0 .05) ,but there was no statistically significant difference between the two groups(P>0 .05) .After 8 weeks′ treatment ,the ef‐fective rate was 91 .7% in ziprasidone groups and 89 .6% in risperidone group .There were no significant differences between the two groups(P>0 .05) .The blood lipids and glucose levels were less increased after ziprasidone treatment ,but was not statistically significant(P>0 .05) .The blood lipids and glucose levels significantly increased after risperidone treatment(P<0 .05) .Conclusion Ziprasidone and risperidone had the same effect on schizophrenia .Ziprasidone had no effect on blood glucose and lipids in schizo‐phrenic patients ,while risperidone could increase blood glucose and lipids level ,we should pay attention to the side effects of long‐term use .