Objective To compare the influence of early weight bearing exercise and plaster fixed method on postoperative function and fracture healing of unstable ankle fracture patients.Methods 50 cases of unstable ankle fracture patients who did surgical treatment in our hospital were selected as the research object,and were randomly divided into an observation group and a control group.The observation group was applied with early postoperative weight bearing exercise,while the control group patients was applied with postoperative plaster fix with 2 weeks.The postoperative function and fracture healing situation of both groups were compared.Results At postoperative 24 hours and 1 week,the perimeter difference of two groups were not statistically different (P＞0.05).At postoperative 6 weeks,the perimeter difference was (0.53 ±0.31) cm in the observation group and (1.25±0.47)cm in the control group,the comparative differences was statistical significance (t=6.39,P＜0.05).The total effective rate of alleviate swelling in the observation group at postoperative 6 weeks was 96.0％,which was higher than that of the control group (76.0％) with statistical difference (x2=4.15,P＜0.05).At postoperative 24 hours,postoperative 1 week and postoperative 6 weeks,the pain index comparison difference between the two groups was not statistically significant (P＞0.05).The Baird-Jackson score of the observation group at postoperative 3 months was not different with the control group with statistically significant (P＞0.05) ; At the postoperative 6 months,the Baird-Jackson score of the observation group was (95.21 ± 8.25),which was higher than that in the control group (82.52±6.96) with statistical difference (t=5.88,P＜0.05).Conclusion The early weight bearing exercise can promote haemal circumfluence,relieve swelling,and be helpful for function recovery and fracture healing of unstable ankle fracture patients.

Objective To construct eukaryotic expressing vector of human melanin-concentrating hormone receptor 1(MCHR1), then to transfect CHO cells with the vector for establishment of stable CHO cell line. Methods The full-length MCHR1 cDNA fragment was amplified by PCR from the human fetal brain cDNA library and then inserted into eukaryotic expression vector pcDNA3.1(+).The recombinant was transfected into CHO cells by lipofectamine TM 2000 after identification of digestion and sequencing on the recombinant eukaryotic expression vector pcDNA3.1(+)/MCHR1. The stable transfected CHO cell line was then established by screening cultures with G418, and the transcription and expression of MCHR1 were identified by RT-PCR, Western blot and immunofluorescence. Results The eukaryotic expression vector pcDNA3.1(+)/MCHR1 was constructed successfully, stable transfected CHO cell line was established, the MCHR1 protein was expressed successfully. Conclusion The construction of eukaryotic expression vector pcDNA3.1(+)/MCHR1 and the establishment of stable transfected CHO cell line provided a solid experimental foundation for further studies on the function of MCHR1.

Objective To construct eukaryotic expression vectors expressing short hairpin RNA(shRNA)sections targeting human MCHR2 and to observe their effects on MCHR2 gene expression in HEK293 cell line.Methods According to the sequence of human MCHR2 gene,the oligonucleotides of shRNA were designed and synthesized and directionally cloned into plasmid pGenesil-1 with enhancing green fluorescence protein(EGFP)gene and Kan gene.The recombinant vectors were confirmed by enzyme digestion analysis and DNA sequencing.The recombinant vectors were transfected into HEK293 cell line by LipofectamineTM2000,the effects on MCHR2 at mRNA and protein levels were observed.Results Four shRNA expressing recombinants and the corresponding negative control vector were constructed and transfected into HEK293 cell successfully.MCHR2 transcript was reduced by about 45.8%-66.4%,the protein of MCHR2 was reduced by about 44.2%-81.0% in four transfectants respectively.Conclusion The construction of eukaryotic expression vectors expressing shRNA sections targeting human MCHR2 and identification successfully established a favourable foundation for further study on the function of MCHR2.

Objective To establish a drug resistant human hepatoma cell line (QGY/DDP) induced by cisplatin (DDP) in vitro, and to investigate its biological features and mechanisms of resistance. Methods The drug resistant cell line (QGY/DDP) of hepatoma was established by intermittent administration of stepwise increas of the dosage of DDP. Drug sensitivity was evaluated by MTT assay. Cell counting was employed to graph the growth curve and to calculate the doubling time. Flow cytometry (FCM) was used to study the cell cycle distribution. In addition, the intracellular platinum accumulation was detected by atomic absorption spectrometry, and the expressions of P-glycoprotein (P-gp) and glutathione S-transferase-? (GST-?) were analyzed by FCM. Results QGY/DDP cell line was established successfully after 3 months with stable resistance to DDP, and the resistance index (RI) was 10.81. The established cell line exhibited obvious cross-resistance to 5-FU, VCR, MMC, EPI and HCPT. Compared with parental cell line, the QGY/DDP cell line grew slower and its doubling time became longer. The proportion of G0/G1 phase cells of the resistant cells was significantly higher than that of their parental cells, whereas the proportion of S and G2/M phase cells was decreased. The cellular platinum accumulation was obviously decreased in the QGY/DDP cell line, and the expression of GST-? in QGY/DDP cells was higher than that of their parental cells, but no over expression of P-gp was found in the resistant cell line. Conclusions QGY/DDP cell line shows the typical and stable resistant phenotype and possesses the basic biological features of resistant cells. The resistance of the cell line to DDP may be due to the reduction of intracellular platinum accumulation and the over expression of GST-?, but may not be related to the expression of P-gp.

Objective To explore the value of combining Uniplate anterior cervical plate system with titanium cage to treat single level cervical spondylotic myelopathy. Method Observing the clinical outcome and X-ray results of 32 patients with cervical spondylotic myelopathy undergone anterior decompression, autograft fusion and internal fixation with Uniplate anterior cervical plate system and titanium cage. Results All patients were followed up for 3 to 12 months. According to bony fusion criteria,no implants breakage, dislocation or loosening was found in all cases in 3 months' postoperatively radiological review. According to Odom classification,excellent outcome was 27 patients, good was 5 patients, the excellent and good rate was 100%(32/32). Conclusions Combining Uniplate anterior cervical plate system with titanium cage to treat single level cervical spondylotic myelopathy has advantages of easy manipulation,safe, less complication and higher fusion rate. It is an ideal way for cervical anterior fixation.

Objective To characterize the x-ray image of vertebral artery type of cervical spondylosis and explore its significance in rehabilitation treatment. Methods The X-ray films of 42 cases of vertebral artery type of cervical spondylosis were measured and analyzed. The treatment approaches including restriction of cervical movement,traction,epidural block and surgery were planned and implemented accordingly. Results The X-ray image revealed the following changes in the patients: the straightened physiologic curvature of the cervical spine,cervical vertebral unsteadiness,zygapophyzeal joint hyperplasia. Various treatment approaches are effective to patients with various manifestations. Conclusion Employing treatment approaches on the basis of X-ray images is of great signi-ficance for the effective treatment of patients with cervical spondylosis of vertebral artery type.

Objective To investigate the value of Q-analysis real-time elasticity in differentiating between benign and malignant thyroid nodules. Methods Eighty-six thyroid nodules in 62 patients with pathologic diagnosis were included in this study and were examined using Q-analysis real-time elasticity. The real-time elasticity features were observed and the quantitative index including the whole elasticity rate and the local elasticity rate were compared between benign and malignant nodules. Results There were 51 benign and 35 malignant nodules according to histopathological examination. The Q-analysis curve of real-time elasticity of benign nodules was smoother and with lower peak, compared with that of malignant nodules. The whole elasticity rate of malignant nodules were significantly higher than that of benign nodules (3.59±0.84 vs 2.32±0.56, P=0.000). And the local elasticity rate of malignant nodules were significantly higher than that of benign nodules (3.96±1.32 vs 2.39±0.58, P=0.000). The cutoff point of whole elasticity rate for the differential diagnosis was 3.25 with sensitivity, specificity and diagnostic accuracy as 71.4%, 96.1% and 86.0% respectively. The cutoff point of local elasticity rate for the differential diagnosis was 3.45 with sensitivity, specificity and diagnostic accuracy as 68.6%, 96.1% and 84.9% respectively. The diagnostic efficiency of whole elasticity rate and local elasticity rate had no significant difference (P=0.591).Conclusions Q-analysis real-time elasticity could provide the real-time elasticity features of thyroid nodules. The whole and local elasticity rate as the quantitative index contributed to the differential diagnosis of benign and malignant thyroid nodules.

Objective To investigate the morbidity and related factors for osteoporosis in postmenopausal patients with type 2 diabetes mellitus (T2DM).Methods The bone mineral density (BMD) at lumbar vertebrae(L1-4), left femoral neck, femoral trochanter and total hip were measured by dual energy X-ray absorptiometry (DEXA) in 79 postmenopansal patients with T2DM.The patients were divided into two groups: osteoporosis group and non-osteoporosis group.The correlations between BMD and age, course of disease, menopausal age, menopausal duration and body mass index (BMI) were analyzed by multivariate regression analysis.Results There were significantly statistical differences in age, BMI, interleukin 6 (IL-6), osteocalcin and menopause duration between two groups.Linear correlation analysis showed IL-6 was positively correlated with osteoporosis (r=0.260, P=0.020) and glycosylated hemoglobin (GHbAlc) (r=0.259, P=0.023) Logistic regression analysis showed that the morbidity of osteoporosis had an independent positive correlation with age, but an independent negative correlation with BMI.Conclusions In postmenopausal patients with T2DM, age and low BMI are independent risk factors for osteoporosis.

Aim To construct a eukaryotic expression vector of human MCHR2 and to analyze the stable expression and signal transduction pathways of MCHR2 in CHO cells. Methods The full-length MCHR2 cDNA fragment was amplified by PCR from the human fetal brain cDNA library and inserted into eukaryotic expres-sion vector pcDNA3.1(+), resulted in the recombinant expression vector pcDNA3.1-MCHR2. The recombinant plasmid was confirmed by the restriction enzymatic digestion and DNA sequencing analysis, and then transfected into CHO cells by lipofectamine. A stably-transfected cell line was obtained by the dominant G418 selection, and the expression of the MCHR2 gene on transcription and translation levels were identified by RT-PCR and Western blot. Signal transduction pathways mediated by MCHR2 were analyzed by measurement of intracellular cAMP and calcium. Results The eukaryotic expression vector pcDNA3.1-MCHR2 was successfully constructed and the MCHR2 gene was stably transfected into CHO cells. A stably-transfected cell line was established and the MCHR2 gene was efficiently transcribed and translated. MCH stimulation had no effect on the production of cAMP, however, it could induce a clear and transient increase of intracellular calcium, suggesting that MCHR2 was only coupled to Gq protein. Conclusion The stable expression of MCHR2 and analysis of its signal transduction pathways in CHO cell line provided a solid experimental foundation for further studies on the function of the MCHR2 gene in vitro.