The special nature of medical care determines the high riskiness of the medical profession that is lacking in other professions. In view of the high-techness and high riskiness of the medical profession, it is only proper to start thinking about setting up corresponding mechanisms for guarding against and dissolving the risks so as to safeguard the legitimate rights of the doctors. Guarding against and dissolving risks of the medical profession fall into a systematic framework of regulations and legal arrangements that are composed of mechanisms for dispersing risks of the medical profession, mechanisms for sharing medical liabilities and systems of medical care and social security.

Objective To investigate the number of osteoclast (OC) precursor in the peripheral blood of patients with ankylosing spondylitis (AS) and its relationship with serum receptor activator of nuclear factor KB-ligand (RANKL) and Osteoprotegerin (OPG) concentration as well as the disease activity. Methods The peripheral blood mononuclear cells from 8 cases of AS patients and 5 healthy controls were cultured in the medium containing macrophage colony-stimulating factor (M-CSF) (25 ng/ml) and RANKL (40 ng/ml). After being cultured for 14 days, cytochemistry was applied to detect tartrate-resistant acid phosphatase (TRAP) expression and the cells with TRAP expression and ≥3 nuclei were counted and defined as OC. Bone resorption assay was used to demonstrate OC function. ELISA was used to measure serum RANKL and OPG concentration in 23 cases of AS and 17 healthy controls. The relationship was analyzed in AS patients between the number of OC precursors and serum RANKL and OPG concentration as well as the disease activity. The indicators of disease activity were Bath ankylosing spondylitis disease activity index (BASDAI), Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). T test, t' test and Spearman correlation were selec-ted. Results ① Significantly higher OC production was observed in the peripheral blood of AS patients than that of healthy control group. The OC number per ten fields was 10.9±3.4 and 6.2±1.3 respectively (P<0.05); ② There was significant difference between AS patients and healthy controls in serum concentration of OPG and RANKL and the ratio of RANKL/OPG. OPG was significantly higher in AS patients [(157±49) pg/ml] than in healthy controls [(105±20) pg/ml] (P<0.05). RANKL was significantly higher in AS patients [(5.4± 3.8) pg/ml] than in healthy controls [(1.6±0.8) pg/ml] (P<0.05). The ratio of RANKL/OPG was significantly higher in AS patients (0.037±0.026) than in healthy controls (0.016±0.008) (P<0.01 );③Significantly positive correlation was observed between the OC number and the serum concentration of RANKL (r=0.692, P=0.009), the ratio of RANKL/OPG (r=0.813, P=0.001);④ In AS patients, serum concentration of OPG was found to have significantly negative correlation with BASDAI (r=-0.444, P=0.044). Serum RANKL concentration was found to have significantly positive correlation with BASDAI (r=0.543, P=0.011). The ratio of RANKL/OPG was found to have significantly positive correlation with BASDAI (r=0.672, P=0.001). Conclusion ① More OC precursors exist in the peripheral blood of AS patients. These cells may differentiate into osteoclasts, which might play a role in joints destructions in AS;② The mechanism of high OC production is likely to be due to high RANKL concentration which is caused by inflammatory reaction.

BACKGROUND:In recent years, minimal y invasive percutaneous plate fixation has been a selectable method to repair fracture of lower limbs, especial y complex fracture of lower limbs. Its advantages are to reduce the damage to soft tissues, and do not destroy bone nutrient supply vessels. However, there is no unified criterion to select which method in the repair of distal tibial fractures. OBJECTIVE:To observe clinical effects of minimal y invasive percutaneous plate fixation versus open reduction and plate fixation in the repair of distal tibial fractures. METHODS:A total of 60 cases of distal tibial fractures treated with minimal y invasive percutaneous plate osteosynthesis (n=35) and open reduction and plate fixation (n=25) were selected. The time of surgery was identified by evaluating soft tissue. We should pay attention to the protection of soft tissue in surgery and reasonable fixation method should be selected. After fixation, we guided patients to do active early functional exercise. They were fol owed up and regularly received X-ray reexamination. Operation time, weight loading time, healing time and functional recovery were observed and the clinical therapeutic effects of the two methods were compared. RESULTS AND CONCLUSION:Al patients were fol owed-up after surgery. They were fol owed up for 3 to 15 months. No significant difference in healing time of type A fracture was detected between minimal y invasive percutaneous plate fixation and open reduction and plate fixation. The healing time of types B and C fracture was better in minimal y invasive percutaneous plate fixation group than in open reduction and plate fixation group. These results indicated that minimal y invasive percutaneous plate fixation in repair of tibial fractures, especial y distal complex tibial fractures, is an effective method. The healing rate of fracture was high, but postoperative complications were less.

Objective To explore the efficacy of intervention combined with traditional Chinese medicine for the treatment of early femoral head necrosis. Methods 62 cases of early femoral head necrosis were treated with interventional and traditional Chinese medicine, all patients were assessed for clinical efficacy before treatment and 6 months after treatment. The clinical hip function, pain index evaluation and rating of the femoral head venography were recorded. Results 41 cases were cured, 18 cases improved,no improvement in 3 cases,the total effective rate was 95. 2%. Clinical hip function after treatment compared with before treatment increased by 32.5%,there was a statistically significant difference (P＜0.01); compared with before treatment, significantly reduced pain (P＜0. 01). After treatment, Ⅲ and Ⅳ were significantly increased number of cases, respectively, compared with the preoperative increase of 146.2% and 160.0%,femoral venography ratings before and after treatment had significant difference(P＜0.01). Conclusion Intervention combined with traditional Chinese medicine treatment could significantly improve the early femoral head necrosis.

AIM: To construct the shuttle plasmid vector for thymidine kinase (tk) and EGFP fusion protein gene driven by IGF - Ⅱ P3 promoter, and investigate the specific killing effect of the HSV - tk/GCV system on hepatocellular carcinoma(HCC) cells in vitro. METHODS: Recombinant shuttle plasmid vector was constructed by techniques of genetic recombination and screening, and identified by restriction digestion and sequencing analysis. Then the recombinant shuttle plasmid was transfected into HepG2 and HeLa cells by techniques of lipofectamine transfection and its expression was detected by fluorescence microscope and RT -PCR. Cell killing after ganciclovir(GCV) application was determined by MTT. RESULTS: Identification of pDC316 -tkEGFP- P3 by enzyme digestion and sequencing analysis showed that the length, inserted location and direction of the target genes which were inserted into the recombinant were correct. It was found that enhanced green fluorescence protein could only be seen in HepG2 cells, but not in HeLa cells. The results of RT -PCR showed that only two bands could be seen in the samples of pDC316 -tkEGFP- P3 transfected HepG2 cells. The MTT test showed the selective cytotoxicity of GCV to the transfected HepG2 cells. CONCLUSION: The shuttle plasmid vector carrying the tkEGFP fusion protein gene driven by IGF - Ⅱ P3 promoter has been constructed successfully and its specific expression in HepG2 cells provided a sound basis for targeted gene therapy for HCC.

AIM:To investigate the effect of RNA interference (RNAi)-mediated insulin-like growth factor 1 receptor ( IGF1R) gene silencing on the growth , migration, and invasion of hepatocellular carcinoma cells .METHODS:The most effective siRNA targeting IGF1R gene was designed and screened .After lentiviral expression vector pLVX-shR-NA2-IGF1R carrying the most effective siRNA sequence was constructed , it was transfected into 293T cells and packed into pLVX-shRNA2-IGF1R lentivirus.Huh7 and Hep3B cells were infected with the pLVX-shRNA2-IGF1R lentivirus to screen the positive clone Huh7 cells and Hep3B cells with the lentivirus .These Huh7 cells and Hep3B cells were cultured to ana-lyze the mRNA level of IGF1R, cell proliferation, cell cycle, cell apoptosis, cell migration/invasion, and the protein levels of IGF1R, Ki-67, p-AKT, p-ERK1, Gli1,β-catenin, cyclin D1, p21 and BCL-XL.RESULTS:The mRNA expression of IGF1R in Huh7 cells and Hep3B cells with pLVX-shRNA2-IGF1R lentivirus was significantly reduced .The proliferation of these cells was remarkably inhibited , and the number in G 1 phase was increased significantly .The percentages of apop-totic cells were increased markedly , and the number of cell migration/invasion was decreased markedly .The protein levels of IGF1R, Ki-67, p-AKT, p-ERK1, Gli1,β-catenin, cyclin D1, p21 and BCL-XL were decreased significantly compared with the blank control group and negative control group .CONCLUSION:The RNAi-mediated IGF1R gene silencing sig-nificantly suppresses the growth and the malignant biological characteristics of Huh 7 cells and Hep3B cells, which may be involved in the reduced protein levels of the above genes induced by down -regulation of IGF1R expression.

OBJECTIVE: To investigate the change of the resistance of common pathogenic bacteria isolated in the First Clinical Hospital of Jilin University during the past 5 years to quinolones.METHODS: Disk diffusion test was used to study the susceptibility test,with the results evaluated based on US Clinical and Laboratory Standards Institute(CLSI).WHONET-5 software was used to analyze the drug resistance data.RESULTS: During 2003～2007,the detection rates of extended spectrum beta-lactamases(ESBLs)-producing Escherichia coli and Klebsiella.spp were 43.6%～60.1% and 51.0%～65.1%,respectively;the detection rates of methicillin-resistant Staphylococcus aureus(MRSA) and methicillin-resistant coagulase negative Staphylococcus(MRSCN) were 50.8%～83.3% and 80.0%～81.5%,respectively;the resistance of common pathogenic bacteria to quinolones showed an increasing tendency,much as in MRSA,MRSCN,feces enterococci,Enterococcus faecalis,and Escherichia coli.CONCLUSION: Monitoring of the resistance of common pathogenic bacteria to quinolones should be strengthened.The change of the antimicrobial resistance should be investigated in order to direct rational drug use in the clinic and prevent the propagation of drug-resistant strains.

OBJECTIVE:To establish the method to identify the polysaccharide element in Holothuria atra Jeager by spectroscopy.METHODS:The purity of polysaccharide in Holothuria atra Jeager was determined by electrophoretic method;the infrared absorption spectrum,ultraviolet absorption spectrum,carbon spectra and hydrogen spectra of NMR were identified in structure by infrared spectrophotometry,ultraviolet spectrophotometry and NMR spectrometry respectively.RESULTS:The gel band after electrophoresis took blue color and the results of spectrum determination accountable for each other.CONCLUSION:Uniformly constituent of polysaccharose was found in the Holothuria atra Jeager,which was an acidic polysaccharose with vitriolic esterfunction,without glucopeptide;?-configuration,?-glucopyranose was found and the intramolecular hydrogen bond was formed by hydroxide radical.

AIM: To clone P1 and P3 promoters of the human insulin-like growth factor Ⅱ(IGF-Ⅱ) gene. METHODS: According to the complete DNA sequence of IGF-Ⅱ gene, the nested primer PCR was performed for amplifying P1 and P3 promoter fragments of the gene from human L-02 cell line.These PCR products were analyzed by agarose gel electrophoresis, and cloned by using TOPO TA Cloning kit. The positive clones containing P1 and P3 fragments were selected and confirmed by sequencing.RESULTS: The DNA sequences of P1 and P3 promoters cloned were accordant with GenBank data. CONCLUSION: In this study P1 and P3 promoters of the IGF-II gene were cloned successfully.

Objective To analyze the structure of microbial community in the bile of patients with obstructive jaundice.Methods From October 2010 to October 2013,117 patients with obstructive jaundice were selected.The results of bile microbial regular culture and anaerobic bacteria culture were both negative.A total of 10 mL bile of each case was aspired and DNA of bile microbial community was isolated.16S rDNA of bile microbial was amplified and underwent terminal restriction fragment length polymorphism (T-RFLP) analysis.Clone libraries were constructed and conducted sequencing and system analysis.Chi-square test was performed for data analysis.Results Among the 117 patients,16S rDNA of 50 cases was positive,and the total positive rate was 42.7 ％.The positive rate of bile bacterial 16S rDNA in stone and tumor cased obstruction was 97.3％ (36/37) and 17.5％ (14/80),and the difference was statistically significant (x2=65.828,P＜0.01).There was no statistically significant difference in the positive rate of bile bacterial 16S rDNA among hilar obstruction,above hilar obstruction and below hilar obstruction,which ware 43.3％ (13/30) and 42.5％(37/87),respectively (P＞0.05).Bile microbial community of obstruction coused by stone was mainly Enterobacteriaceae (Escherichia,Salmonella,Klebsiella,Proteus),Streptococcus (Streptococcus),digestive coccaceae (digestive bacteria genus,peptostreptococcus),Micrococcus branch (Staphylococcus),Propionibacterium (Propionibacterium) and Neisseriabacteria (Acinetobacter).Bile microbial community of obstruction caused by tumor was mainly Enterobacteriaceae (Clay Beth spp,Escherichia,Salnonellatyphi) and Micrococcus branch (Staphylococcus).Conclusion The condition and variety of bile microbial community of patients with obstructive jaundice could be effectively evaluated by 16S rDNA fragment through T RFLP analysis.