Objective To compare the clinical results of Endobutton and Rigidfix graft fixation at the femoral tunnel after ACL reconstruction with hamstring autografts.Methods 48 patients accepting ACL reconstruction with hamstring autografts were retrospective studied.According to different fixation adopted at the femoral tunnel,they were divided into group A(24 patients,with Rigidfix cross pin)and group B(24 patients,with Endobuaon).All Cases were fixed with Intrafix interference screw and Spiked Washer screw at the tibial tunnel.The average follow-up time was 27 months.KT-1000 arthrometric measurement and the IKDC,Lysholm,Tegner scores were used for evaluation.The diameter of bone tunnel was also measured by MRI.Results Tunnel widening were found in both groups,but the increase of both the femoral and tibial tunnels in group B were significantly greater than group(P<0.05).In both groups after operation,there was significantly increases of IKDC suhjective scores,Lysholm scores,Tegner scores and KT-1000 results also had obviously improvement(P<0.05).But no significant difference was found between two groups after operation.Conclusion Compared with double cross-pins for fixation at the femoral tunnel,tunnel widening was more prone to happen when using Endobutton suspensory fixation system.

Objective To discuss the clinical value of serum tumor specific growth factor (TSGF) and vascular endothelial growth factor (VEGF) in early diagnosis of thyroid cancer.Methods Eighty thyroid cancer patients (thyroid cancer group),120 nodular goiter patients (nodular goiter group),and 80 healthy adult (blank control group) were enrolled in this study.The level of TSGF and VEGF in two groups were detected by ELISA and compared.Results The level of TSGF and VEGF in thyroid cancer group and nodular goiter group were significantly higher than those in blank control group (P ＜ 0.05).Conclusion Serum TSGF and VEGF have some clinical value in early diagnosis of thyroid cancer.

Objective To evaluate the effects of recombinant human bone morphogenetic-2-polylactide sustained release nanospheres (rhBMP-2-PLA-Ns) on cultured rabbit osteoblasts in vitro. Methods Rabbit osteoblasts were cultured in vitro, and rhBMP-2-PLA-Ns was added into the medium of the third generation of rabbit osteoblasts. The expression of the proliferating cell nuclear antigen (PCNA) was examined by immunofluoreacence staining, and the formation of tuberculums observed with alizarin red staining. Western blot was used to detect the effects of rhBMP-2-PLA-Ns on the expression of vascular endothelial growth factor (VEGF), which was compared with that in rhBMP-2 group and blank group. Results There was no significant difference in the number of osteoblasts with positive PCNA expression among three groups five days later. Ten days later, the number of osteoblasts with positive PCNA expression in rhBMP-2-PLA-Ns group was significantly higher than that in rhBMP-2 group and blank group, which indicated that rhBMP-2-PLA-Ns could enhance the expression of PCNA. Compared with rhBMP-2 group and blank group, rhBMP-2-PLA-Ns could significantly enhance the formation of tuberculums, with statistical difference (P<0.05). The expression of VEGF was detected in all three groups, and the level of the VEGF expression in rhBMP-2-PLA-Ns group was significantly higher than that in the other 2 groups (P<0.05). Conclusion The biological activity of rhBMP-2-PLA-Ns is superior than that of rhBMP2, and rhBMP-2-PLA-Ns can promote the proliferation, mineralization of osteoblasts and the secretion of VEGF, which has a better prospect in facilitating traumatic bone healing.

It expounded the important position of physique in etiology and process of disease,investigated essence and application of physique in disease control,revealed the classifi cation of physique in three yin three yang.Also it expounded the signif icance of diagnosis of disease in TCM theory and clinical therapy,then revealed basic pathogenesis of diabetes:damaged yin and qi due to inner heat.Then it discussed the syndrome differentiation in the TCM treatment and suggested that it should combine with physique differentiation and diagnosis of disease.At last it expounded the trinity of physique differentiation,diagnosis of disease and syndrome differentiation in treating diabetes and its complication.

OBJECTIVE: To study the how's for pharmacists at inpatient pharmacy to provide pharmaceutical care for improved safe and effective use of drugs. METHODS: Pharmacists at inpatient pharmacy can provide pharmaceutical care as per specific clinical needs, and start from such details as preparing the booklet of commonly-used chemotherapy drugs, offering face-to-face advises on drug use for diabetics, and helping nurses to manage the medicine cabinets. RESULTS & CONCLUSION: Pharmacists at inpatient pharmacy should provide the right pharmaceutical care that can meet actual clinical needs.

ObjectiveTo develop an antibody-array system for multiple detection of antibodies against Japanese B encephalitis virus (JEV),Tick-borne encephalitis virus (TBV),Dengue virus ( DENV ),West Nile virus (WNV),Western equine encephalitis virus (WEEV) and East Equine encephalitis virus (EEEV).MethodsRecombined antigens were spotted on array as capture antigens.Specific antibodies were detected by using a sandwich ELISA format.Rabbit antiserum was employed to select and confirm the specificity of antigens and to optimize the conditions of the assay.The detection efficiency of the system was validated by 40 clinical suspected serum samples and compared with the relative ELISA assays.ResultsEleven recombined antigens were selected as diagnostic antigens with high specificity.Better detection could be achieved when scale of antigen concentrations were within 0.125-0.900 mg/ml and the serum dilutions were 1:100-1:1000.When detecting the 26 clinical suspected TBE serum samples,20 were IgG positive (76.9％),and 17 were IgM positive (65.3％) which was 96.1％ and 84.6％ consistent with the relevant ELLSA tests,the 8 clinical suspected JEV serum samples,4 were IgG positive (50.0％),and 5 were IgM positive (62.0％),which was 86.3％ and 90.1％ consistent with the relevant ELLSA tests.As for the 22 DEN serum samples,13 were IgG positive (60％) and 15 were IgM positive (68％) which was 85％ and 93％ consistent with ELISA.The specificity of the assay was 100％ and the sensitivity was higher than the relative ELISAs.ConclusionThe developed antibody-array is highly specific and reliable,which could be used for the detection of antibodies against the 6 arboviruses.

Objective To express and purify the recombinant nucleoprotein fragments of hemor-rhagic fever with renal syndrome ( HFRS) virus and to evaluate their diagnostic efficacy by using array-ELISA technology .Methods The target genes encoding nucleoprotein fragments of HFRS virus were amplified by PCR, and then inserted into prokaryotic expression vectors to construct the recombinant plasmids of pET -32a (+)/Pn and pET-32a(+)/Pc.The plasmids were transformed into E.coli BL21 ( DE3) to induce the ex-pression of nucleoprotein fragments by IPTG and the expressed products were purified by affinity chromatog -raphy using Ni-NTA agarose.The specificity and sensitivity of the recombinant antigens were evaluated by the assay of array-ELISA using commercial colloidal gold assay kit as a comparison .Results The recombi-nant nucleoprotein fragments of HFRS virus were correctly expressed in E.coli and highly purified by affinity chromatography .Array-ELISA showed that 13 of 16 suspected serum samples were positive by using the His-Pn protein as diagnostic antigen , consistency with the commercial colloidal gold assay kit reaching 94%. Conclusion The recombinant His-Pn protein expressed in E.coli cells could be used for specific serodiag-nosis of HFRS virus as its high antigenicity and sensitivity .The array-ELISA is an effective assay for the de-tection of virus at protein level .

Objective To establish a one-step recombinase polymerase amplification (RPA) method for pathogen screening and rapid detection in the field targeting for five hemorrhagic fever related viruses (Zaire ebola virus, Sudan ebola virus, Marburg virus, Lassa virus and Yellow fever virus). Methods The specific nucleic acid (NA) fragments of each virus were selected as target genes by genome sequence analysis, and the primers and probes for RPA assays were designed according to the sequence. A series of diluted template genes were used for RPA detection to determine the sensitivity. The hemorrhagic fever-related viral nucleic acids were used for RPA detection to determine the specificity. The amplification experiments were carried out at different temperature ranging from 37℃ to 42℃ to validate the reaction temperature range. Results The RPA reaction systems of the five hemorrhagic fever viruses could effectively amplify the target genes, the sensitivities were between 1.5×102 and 1.5×103 copies. No cross reactions existed with the other hemorrhagic fever-related viral genes. Meanwhile, RPA assay could effectively amplify the target genes at 37-42℃. Conclusion The isothermal RPA assays of five hemorrhagic fever viruses are established, which may amply target genes fast and react at a wide temperature range, and be potentially useful for in field pathogens detection.

Objective To explore the relationship of DVL2 expression and the development of (CCRCC) by comparing the changes of DVL2 mRNA and protein expression in CCRCC specimens and matched normal renal specimens and its clinical significance. Methods DVL2 mRNA expressions in 22 CCRCC tissues, the matched adjacent normal tissues, and 10 CCRCC tissues alone were examined by semiquantitative RT-PCR and fluorescence quantitative PCR (real-time RT-PCR). Meanwhile, the different expression of the CCRCC between TNM Stage Ⅲ + Ⅳ and Stage Ⅰ +Ⅱ was also examined. Furthermore,immunohistochemistry was employed to examine DVL2 protein expression in 22 CCRCC and the matched adjacent normal tissues, and the other 10 CCRCC tissuses without the matched tissues. Results The DVL2 mRNA expression levels in 17 CCRCC tissues were increased by semi-quantitative RT-PCR and by real time RT-PCR compared with that in corresponding adjacent normal tissues, with the difference being significantly different (t = 2.535, P =0.0197). The DVL2 expression of 8 in 13 Ⅲ + ⅣCCRCC was higher than Ⅰ +ⅡCCRCC. Immunohistochemical examination showed that the DVL2 protein was located in cytomembrane and cytoplasm. Moreover, the positive level of DVL2 protein in CCRCC tissues[81.8 % (18/22)]was significantly higher than those in the adjacent tissues. However the expression was not associated with patients' age, gender, TNM stages (Fisher exact frenquently, P >0.05). Conclusion The DVL2 expression in CCRCC is obviously higher than the corresponding normal tissues in the level of mRNA and protein. And the higher DVL2 expression might be closely associated with the development and progression of CCRCC in the level of mRNA, which may be a potential molecular marker of CCRCC development and metastasis mechanism.

Objective To express and purify Japanese encephalitis virus ( JEV) EDⅢprotein and evaluate the possibility of using it as a candidate antigen in JEV diagnostic kit .Methods PCR primers spe-cific for the gene encoding JEV EDⅢprotein were designed and used to amplify the gene fragment by RT-PCR.The cloned gene fragment was then inserted into pET-30a (+) to construct the recombinant expression plasmid.The transformed E.coli BL21 carrying expression plasmid were induced by IPTG to express JEV EDⅢ protein.The expressed JEV EDⅢprotein and a control antigen of tick-borne encephalitis virus protein were deposited in small spots to set up ELISA microarray .The serum samples from patients with Japanese encephalitis and healthy people were detected by Array-ELISA.The results obtained by Array-ELISA were compared with those by using indirect immunofluorescence assay .Results The gene fragment encoding JEV EDⅢprotein was successfully cloned and expressed in E.coli BL21.The recombinant protein could be used in Array-ELISA assay for the detection of serum samples from patients with Japanese encephalitis and healthy subjects .The results were consistent with those by using indirect immunofluorescence assay.Conclusion The recombinant JEV EDⅢprotein can be used as a candidate antigen for the diagnosis of JEV infection .