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.

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.

A large series of lens were collected from normal and senile cataractous human eyes, and were investigated according to the theories of oxyradical and lipid peroxide (LPO). The findings showed that the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase in the senile cataractous lens were very significantly lower than those in the normal ones, while the content of malondialdehyde (MDA) in the lens of senile cataractous eyes was very significantly higher than that of normal eyes. The results suggest that the oxyradical and LPO might be one of the direct causes leading to senile cataract formation.

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 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.

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 develop and identify monoclonal antibodies (McAbs) against adult worm of Angiostrongylus cantonensis and observe its applicability. Methods BALB/c mice were immunized with soluble antigen of adult worms of A.cantonensis. The spleen cells of immunized mice were fused with myeloma cell, and the hybridoma secreting high titer of McAbs with high specificity was screened. By using the McAbs, serum of angiostrongyliasis patient and sera of the rats infected with A.cantonensis were detected by Western blotting and double antibody sandwich ELISA respectively. Results Three McAbs were established (2A2,3F1,4H2), which all showed no cross reaction with antigens of Schistosoma japonicum, Paragonimus westermani, Cysticercus cellulosae and Trichinella spiralis. Western blotting analysis demonstrated that the three McAbs recognized a Mr 15 000 soluble antigen of adult worm of A.cantonensis and recognized the Mr 24 000 and Mr 15 000 circulating antigens from the serum of angiostrongyliasis patient. The double antibody sandwich ELISA detection showed a positive rate of 76^5%. Conclusion Three hybridoma cell lines against adult worm of A.cantonensis have been established which secret high titer of McAbs with high specificity and seem promising in detecting the circulating antigen of the angiostrongyliasis patient.

AIM: To investigate the genetic and epigenetic alterations of p14~ ARF gene and mutation status of p53 gene in human primary colorectal carcinomas and to analyze the relationship between the two gene changes and the role of abrogation of the p14~ ARF -p53 pathway in colorectal carcinogenesis. METHODS: The homozygous deletions, mutations, methylation of 5′ CpG islands, mRNA expression of p14~ ARF gene and mutations of p53 gene were assessed by PCR, direct sequencing, methylation-specific PCR, and RT-PCR in the tumorous and matched adjacent normal colorectal tissues from 56 patients with colorectal carcinoma. RESULTS: ① p14~ ARF alterations were detected in 27% (15/56) of colorectal carcinoma tissues studied, of which 1 case showed homozygous deletion, 14 cases showed 5′ CpG island methylation, and no mutation was found in any tumor. ②15 colorectal carcinomas with p14~ ARF alterations indicated lack of (13 cases) or at low level of expression (2 cases) of p14~ ARF mRNA, while expression of the p14~ ARF transcript was detected in the remaining 41 colorectal carcinomas and any matched adjacent normal colorectal tissues. ③ The mutations of p53 gene were detected in 48% (27/56) of colorectal carcinomas investigated. ④ Of these 56 cases, 12 had p14~ ARF alterations alone, 24 had p53 mutations alone, 3 had both p53 mutations and p14~ ARF methylation, and 17 had neither. 70% (39/56) of the samples had either or both abnormalities of the two genes, and p14~ ARF hypermethylation was related to wildtype p53 (P

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.