Human astrovirus (HastV) is recognized as one of the leading causes of acute viral diarrhea in infants. The HastV non-structural protein, nsPla, and C-terminal protein, nsPla/4, contain various conserved functional domains,and may play an important role in virus replication, transcription and the virus-host interactions of HastV. This study used an E. coli system to investigate the expression of nsPla and nsPla/4 proteins. Firstly,the nsPla and nsPla/4 genes of HAstV-1 were cloned into the prokaryotic expression vector,PGEX-4T-1, to build the PGEX-4T-1a and PGEX-4T-la/4 fusion protein plasmids. Then, the recombinant plasmids were transformed into Escherichia coli BL21 (DE3) and induced with isopropyl-β-D-thiogalactopyranoside (IPTG). The optimal expression conditions of the two fusion proteins were identified and then analyzed by polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting, respectively. The results showed that the pGEX-4T-la fusion protein was maximally expressed at 30 °C after 12 hours of induction with 1.0 mM IPTG. The pGEX-4T-la/4 fusion protein was maximally expressed at 20 °C after 8 hours of induction with 0.5 mM IPTG. Western blot analysis showed that the two fusion proteins specificity reacted with the anti-nsPla and anti-GST monoclonal antibodies, respectively. This study successfully obtained the HAstV non-structural protein, nsP1a, and its C-terminal protein nsP1a/4 protein using an E. coli system. This novel study lays the foundation for future research into the pathogenic mechanisms of human astrovirus and the functions of its non-structural protein.
Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Mamastrovirus ; genetics ; metabolism ; Viral Nonstructural Proteins ; genetics ; metabolism

In this paper, a novel and simple RP-HPLC method for the determination of related substances of tiopronin for injection was described. The RP-HPLC analysis was performed on a C18 column, with acetonitrile-0. 1% phosphoric acid(8 ∶92), mobile phase in isocratic mode at a rate of 1. 0 mL/min. The photodiode array detector was set at 210 nm. Seven related substances were detected and the structures were characterized by mass spectrometry. The method showed great suitability, specificity and excellent linearity over the concentration range of 0. 3 to 50 μg/mL(r≥0. 999), and the limits of detection and quantitation were found to be 0. 10 and 0. 31 μg/mL, respectively. The accuracy of the method determined by the entire mean recovery ranged from 98. 7% to 103. 7%. The intra-and inter-day precision was satisfactory(RSD≤4. 4%)and robust(RSD≤6. 4%). And this method was successfully applied for the determination of related substances of tiopronin for injection, which revealed the retention of sulfhydryl compounds and glycine analogues on the RP-HPLC and the effect of the pH value of the mobile phase on the chromatographic behavior of the analytes.

In order to strengthen the training of grassroots health personnel,in Shanghai the program of general practice training has been included in the residency training systemsince 2010,and the program integrated theresidency training with master degree study.At the same time the training of general practitioners in Shanghai also included the following programs:job-transfer training for grassroots doctors,order oriented training for rural doctors,order oriented free medical education,and standardized training of assistant general practitioners.

Objective To explore the effects of treatment decision aid on addressing decision-making difficulties and anxiety, depression in patients. Methods Totally 289 patients with lung cancer who were treated in Harbin Medical University Tumor Hospital between July and December 2016 were selected by purposive sampling and randomly divided into treatment group (n=147) and control group (n=142). Patients in the control group received routine guidance and health education from doctors, while patients in the treatment group received decision support manuals and took part in treatment decision-making with medical staff. The Decision Distress Scale and the Hospital Anxiety and Depression Scale were used to assess the intervention effects. Results There was no statistically significant difference between the two groups judging from the scores of the Decision Distress Scale before the intervention (t=0.237, P=0.532). After the intervention, the patients in the treatment group scored lower than the patients in the control group (t=6.437, P<0.001), and there was no statistically significant difference in the level of anxiety (t=0.214, P=0.617) and the level of depression (t=0.105, P=0.874) between the two groups. Conclusions Treatment decision aid can address the decision-making distress and mitigate the sense of uncertainty in patients effectively.

Rheumatoid arthritis(RA)is an autoimmune disease with the basic pathological manifestation of synovial inflammation.Symmetric poly-articular pain and swelling are the main symptoms in clinical practice,and even extra-articular manifestations and comorbidities such as interstitial fibrosis and coronary artery disease are triggered,which seriously affect the quality of life of patients.Traditional Chinese medicine(TCM)has achieved good clinical efficacy in the prevention and treatment of RA with the advantages of multi-pathway,multi-target,multi-component,and less toxic side effects,and plays an important role in the treatment of RA.Recently,many studies have demonstrated that Chinese medicine monomers and Chinese herbal compound can control inflammation,reduce angiogenesis,induce apoptosis of synovial fibroblasts,and inhibit their proliferation,invasion and migration by regulating the PI3K/Akt signaling pathway,so as to play a key role in the treatment of RA.For this reason,the article summarizes current knowledge regarding the PI3K/Akt signaling pathway and its role in RA,as well as summarizes the current research progress of TCM in the treatment of RA by regulating the PI3K/Akt signaling pathway.The aim of this review is to provide theoretical bases for the prevention and treatment of RA and the development of new drugs.

Ankylosing spondylitis(AS)is a chronic inflammatory autoimmune disease characterized by inflammatory back pain.Its pathological features mainly include inflammation,bone destruction,and pathologic new bone formation.The etiology of AS is complex,and it may be related to genetics,infections,the environment,and intestinal flora.Its pathogenesis has not yet been clarified.In recent years,osteoimmunology,as a new theme in the study of inflammatory arthritis,plays an important role in the pathogenesis and development of AS,which was embodied in the inflammatory response and imbalance of bone metabolism.Traditional Chinese medicine(TCM)has the characteristics of multiple pathways,multiple components,multiple targets and multiple levels.TCM can improve the inflammatory response and bone metabolism imbalance of AS by regulating the osteoblasts of the skeletal system and the related factors of the immune system,thus to prevent and control AS.For this reason,the paper summarizes the role of bone immunology in the pathogenesis of AS,and reviews the current status of research on the intervention of TCM in bone immunology for the treatment of AS,with a view to providing certain references for the future clinical application of TCM in the prevention and treatment of AS.