Objective To clone and fuse the cDNA of human cytokeratin 9 in prokaryotic expression system,and purify and identify the fusion protein.Methods The cDNA fragment of human cytokeratin 9 was amplified from human keratinocyte (HaCaT) total RNA with specific primers.The PCR products were cloned into vector pET-28a,then the fusion protein of his-CK9 was induced by IPTG.The expressed fusion protein of his-CK9 was purified by nickel ion affinity chromatography and identified by SDS-PAGE and Western blot.Results The sequencing proved that the recombinant vector of the cDNA of CK9 was correct.The fusion protein of his-CK9 was induced to be expressed in E.coli.The fusion protein of his-CK9 was highly purified and his-CK9 showed specific binding to the commercialized antibodies of CK9.Conclusion The recombinant vector of pET-28a-CK9 has been successfully constructed,and the fusion protein of his-CK9 has been successfully expressed and purified.

Objective To evaluate the effect ofJiawei-Danggui-Buxue decoction in the treatment for the patients with hemorrhagic anemia after total knee arthroplasty (TKA).Methods A total of 63 TKA patients with hemorrhagic anemia were included, and were divided into the treatment group (31 patients) and the control group (32 patients). All patients started to take medicine the second day after surgery. The control group took the Ferrous Succinate Tablets and the treatment group took Jiawei-Danggui-Buxue decoction in the following 4 weeks. The Hb and Hct were detected,and the changes of TCM syndrome scores and the total effect rates of anemiawere observed.Results The total effect rate of TCM syndrome scores in the treatment group was 93.54%, which was significantly higher than control group 83.87% (P<0.05); The total effect rate of anemia in the treatment group was 90.32%, which is significantly higher than control group 75.00% (P<0.05). At the fifteenth day after operation,. And at the postoperative 29th day, the levels of Hemoglobin (129.61 ± 13.02 g/Lvs. 113.35 ± 15.46 g/L,t=6.245) and Hematocrit (0.45% ± 0.03 %vs. 0.41% ± 0.04%,t=5.962) in the treatment group were significantly higher than those in the control group (P<0.05).Conclusions TheJiawei-Danggui-Buxue decoctionimprovethe Hb, HCT and the clinical effect rates of anemia and TCM syndromes for the patients with hemorrhagic anemia after TKA.

With the common occurrence of knee osteoarthritis, total knee arthroplasty has become an effective method for the treatment of middle and late knee osteoarthritis. Continuous passive movement is an effective auxiliary method for rehabilitation after total knee arthroplasty, but it also has some limitations. This article reviews the research progress of continuous passive movement after total knee arthroplasty.

Bone infection is a serious infectious disease in clinical practice due to its difficult treatment and poor prognosis. Therefore, early accurate diagnosis of bone infection is the key for successful treatment. The traditional detection methods is time consuming with lower positive rate. Therefore, it is urgent to find better detection techniques to identify the pathogenic microorganisms of bone infection. In recent years, metagenomic second-generation sequencing technology has been widely used in the diagnosis of clinical infectious diseases because of its advantages of accurate, rapid, efficient and comprehensive diagnosis of pathogenic microorganisms, and can be used as an effective tool for the diagnosis of bone infections. This paper mainly reviews the advantages, disadvantages and development direction of metagenomics next-generation sequencing technology in the diagnosis of bone infection.