OBJECTIVE:To establish a method to determinate the content of paeoniflorin in Bailing tablet,and to evaluate the uncertainty of results. METHODS:HPLC was performed on the column of Platisil C18 with mobile phase of water- acetonitrile(87:13,V/V) at a flow rate of 1 ml/min,the detection wavelength was 230 nm,the column temperature was 25 ℃ and the injection volume was 10 μl. Quantitative analysis was conducted for the processes and procedures and the uncertainty was evaluated. RESULTS:The linear range of paeoniflorin was 0.100 1-2.502 8 μg;RSDs of precision,stability and reproducibility tests were lower than 2.0%,recovery rate was 98.7%-100.9%(RSD=0.86%,n=6). Reference substance amount,elution produce and the accuracy of method had great effect on the uncertainty. CONCLUSIONS:The established method can be used for the content determination of paeoniflorin in Bailing tablet,and the uncertainty evaluation provides reliability for the method.

AIM: To observe the curative effect of titanium wiring in treatment of the fractures of Neer Ⅱ type distal clavicle and dislocation of Tossy Ⅲ type acromioclavicular joint. METHODS: From March 2004 to December 2006, 37 cases with Neer Ⅱ type distal clavicle fractures or Tossy Ⅲ type acromioclavicular joint dislocation were treated with titanium wiring in Changzheng Hospital, Second Military Medical University of Chinese PLA. Titanium wiring (produced by Sofamor-Danek Company in USA) had good histocompatibility, intensity of tension and flexibility. The broken coracoclavicular ligament was repaired directly by titanium wiring in all cases. The outcomes were evaluated according to Herscovici's criteria. RESULTS: All the cases were followed up for 6 to 24 months. All the dislocated acromioclavicular joints returned to normal 2 weeks after operation. Excellent and good rate of shoulder function was 100% according to Herscovici standard. X-ray imaging showed all clavicle fractures united, acromioclavicular joint had no redislocation or titanium wiring breaking. The incisions of all cases had no obvious swelling and exudation. CONCLUSION: When Neer Ⅱ type distal clavicular fracture and Tossy Ⅲ type acromioclavicular joint dislocation are treated with titanium wiring, the acromioclavicular joint can exercise earlier and recover quickly.

Objective To evaluate the prevalence and risk factors of peripheral arterial obstructive disease(PAD) in type 2 diabetes mellitus(T2DM) patients. Methods A total of 401 patients with T2DM(201 men and 200 women),mean aged was(63.4?8.8)years.PAD was diagnosed by an ankle-brachial index(ABI)≤0.9 on either leg.Results Overall prevalence of PAD was 19.4%,without significant sexual difference.Univariate analyses disclosed age,hypertension,systolic blood pressure(SBP),low dense lipoprotein cholesterol(LDL-C) and T2DM duration as significant risk factors.In multiple Logistic regression,age,SBP,LDL-C and T2DM duration were independent risk factors with respective odds ratios(95% confidence intervals,CI) of 1.94(1.20-2.60),1.12(0.88-1.43),1.32(0.97-1.63) and 1.27(0.96-1.73) respectively.Conclusions Prevalence of PAD in T2DM patients is(19.4)% and the major risk factors were older age,higher SBP,higher LDL-C and T2DM duration.This implies that it is very important to prevent the incidence of PAD for better glucose,lipid and blood pressure control early.

Objective To investigate the distribution of alpha2-HS glycoprotein (AHSG) gene polymorphisms and the relationships of AHSG gene polymorphisms with atherosclerosis as well as serum bone related biochemical mark-era. Methods Blood samples of 344 hospitalized female patients, aged 20 ～ 80 years, were sampled for serum bone alkaline phosphatase, cross-linked N-telopeptide of collagen type Ⅰ, cross-linked C-telopeptide of collagen type Ⅰ , osteoprotegrin and leptin were determined by ELISA. Serum TC,TG and calcium content were detected. Poly-morphism of AHSG gene was detected by polymerase chain reaction fragment length polymorphisms (PCR-RFLP) of restriction enzyme Sac Ⅰ. BMD (Norland XR-36) of the anteroposterior spine (AP), supine lateral spine (Lat) and femoral neck (FN) were measured. Morphological changes in the aorta and bone of type GG patient were detected by pathological microscopy. IMT were measured by color doppler ultrasound equipment(SEQUOIAS12). Results (1) The genotype frequency of CC, CG, and GG were 59.7%, 25.1% and 15.2% respectively in all elderly female patients. There were significant difference in blood lipids, Ca~(2+) and serum bone relative biochemical markers to different AHSG genotypes. (2)There were significant differences in the BMD of the AP, Lat, FN and IMT and the serum biochemical markers among the CC, CG and GG genotypes. (3)GG-female patients bone tissue pathology section verify the AHSG polymorphism genetic mutation and atherosclerosis, osteoporosis development of the relationship. Conclusion There was close relationship among AHSG polymorphism variation and the incidence of arteriosclerosis and osteoporosis in elderly female.

Objective To investigate the distribution of alpha2-HS glycoprotein(AHSG) gene polymorphisms and the relationships of AHSG gene polymorphisms with atherosclerosis as well as serum bone related biochemical markers.Methods Blood samples of 344 hospitalized female patients,aged 20～80 years,were sampled for serum bone alkaline phosphatase,cross-linked N-telopeptide of collagen typeⅠ,cross-linked C-telopeptide of collagen type Ⅰ,osteoprotegrin and leptin were determined by ELISA.Serum TC,TG and calcium content were detected.Polymorphism of AHSG gene was detected by polymerase chain reaction fragment length polymorphisms(PCR-RFLP) of restriction enzyme Sac Ⅰ.BMD(Norland XR-36) of the anteroposterior spine(AP),supine lateral spine(Lat) and femoral neck(FN) were measured.Morphological changes in the aorta and bone of type GG patient were detected by pathological microscopy.IMT were measured by color doppler ultrasound equipment(SEQUOIA512).Results(1) The genotype frequency of CC,CG,and GG were 59.7%,25.1% and 15.2% respectively in all elderly female patients.There were significant difference in blood lipids,Ca2+ and serum bone relative biochemical markers to different AHSG genotypes.(2)There were significant differences in the BMD of the AP,Lat,FN and IMT and the serum biochemical markers among the CC,CG and GG genotypes.(3)GG-female patients bone tissue pathology section verify the AHSG polymorphism genetic mutation and atherosclerosis,osteoporosis development of the relationship.Conclusion There was close relationship among AHSG polymorphism variation and the incidence of arteriosclerosis and osteoporosis in elderly female.

Objective To investigate the mitochondrial energy metabolism in D-galactoseinduced cell ageing model.Methods MRC-5 cells were cultivated for 72 hours in a medium containing 55 mmol/L D-galactose.The analysis of cell proliferation capacity by CCK8 method,β-galactosidase staining and detection of p21 protein expression level were performed for identifying cell senescence.The cell oxidation-reduction state was evaluated by an analysis of cellular ROS levels,SOD activity,MDA content and oxidative damage level of mitochondrial DNA(mtDNA).For purpose of detecting mitochondrial function and its impairment,mitochondrial morphology was observed by electron microscope,mitochondrial quantity was analyzed by flow cytometry,mitochondrial membrane potential(△Ψm) was measured by JC-1 staining,and ATP content was analyzed by HPLC,and mitochondrial oxygen consumption rate was detected by Seahorse cell energy metabolism detection system.Results The decreased MRC-5 cell proliferation,up-expression of p21 protein,increased β-galactosidase activity were observed in D-Gal-treated cells,which indicated the cell premature senescence.When treated with D-Gal,the significantly increased ROS and MDA level,decreased SOD activity and increased oxidized mtDNA proved that the cells kept higher oxidative stress.D-Gal induced-mitochondrial impairment was evidenced by the dimming of mitochondrial cristae and double membrane structure,decrease of transmembrane potential and ATP synthesis,and decrease of its oxygen consumption rate(OCR).Conclusions The 55 mmol/L D-Gal causes an impairment of mitochondrial structure and a decrease of function of energy metabolism,which is associated with cellular senescence induced by D-Gal.

Objective To discuss the outcome of repair of limbs bone defect by using mineralized collagen graft.Methods The clinical data of 35 cases who diagnosed the existence of bone defect from January 2013 to December 2013 in our department were retrospectively analyzed.After operation,Lane Sandhu X-ray score standard was evaluated.Results All patients were followed up for 12 months.Postoperative 1 month,Lane Sandhu X-ray score was 3 points in 20 cases and 15 cases with 2 pointss;postoperative 6 months,Lane Sandhu X-ray score was 9 points in 21 cases,10 cases with 8 points and 4 cases with 7 points.Postoperative 12 months,Lane Sandhu X-ray score was 12 points in 28 cases,7 cases with 11 points.The scores of postoperative 6 months were better than the scores of postoperative 1 month [(2.60±0.49)points vs.(8.49±0.12)points,t=107.860,P=0.000].There was significant difference between the scores of postoperative 12 months and scores of postoperative 6 months [(8.49±0.12)points vs.(11.8±0.06)points,t=41.630,P=0.000].Conclusion MC is used as an effective bone substitute material,and its clinical effect is good.

New field battle X-ray vehicle(model XCY2002-1/200) is used usually and control method is grasped too. By analyzing operation capability, the advantages and disadvantages are found, and then corresponding improvement is proposed to educe efficiency of medical support.

Objective To investigate the changes between type 2 diabetes mellitus(T2DM) with peripheral vessel disease and osteoporosis. Methods A total of 100 cases T2DM patients were divided into non-peripheral vessel disease (A group) and peripheral vessel disease (B group). B-mode ultrasonography to measure the intima-media thickness(IMT) of carotid artery,dual energy X-ray absorptiometry (DEXA) to measured bone mineral density(BMD) and VS-1000 arteriosclerosis diagnometer to measure brachi-ankle pulse wave velocity (baPWV) were used in 100 T2DM patients. The clinical data and BMD were compared between two groups. Results Compared with normal group,the BMD in T2DM patients significantly decreased.Compared with T2DM patients without peripheral vessel disease,the BMD decreased in T2DM patients with peripheral vessel disease. Conclusions Atherosclerosis in T2DM with peripheral vessel disease is probably related to osteoporosis, and the relationship between them is worth further investigation.