ObjectiveTo construct expressing vector of microRNA with molecular cloning methods which target CD4 and electroporating the vector to the MT4 cell to determine its effect to CD4 expression.MethodsPredict a microRNA can target CD4.Linking the pre-mir-181a PCR products to T vector,then cloning into the pEGFP-N1 vector after enzyme digestion.To test the integrity through the colony PCR and sequencing analysis.Electroporating the vector to MT4 cell,using FACS to test the CD4 expression.ResultsHsa-mir-181a is able to target CD4.The sequence of the construct was correct.The hsa-mir-181a is stable expressing in MT4 after electroporating with the vector and MT4 cell CD4 was down-regulated.ConclusionThe construct can be stable expressing hsa-mir-181a in MT4 cell and down-regulating the CD4 expressing.This method can be utilized as a novel intervention to the HIV fusion,it shows potential as a gene therapy tool in vitro.

BMP6 is a potent protein for future treatment strategies of bone regeneration as it is a very important regulator of bone homeostasis. Active BMP6 is a dimer containing multidisulfide bonds and is a highly hydrophobic protein prone to aggregation. To obtain soluble and active BMP6 in Escherichia coli, we compared the effects of four N-terminal fusion tags (TRX, GST, MBP and CBD) and N-terminal His6-tag. The expression and solubility were tested under the different conditions (expression hosts, temperatures and inductor concentrations). A series of experiments leads to the finding that the placement of MBP before the BMP6 is best in availing the soluble expression of the protein. Our study alsodemonstrates that in E. coli BL21trxB(DE3) cytoplasm, which is a thioredoxin reductase mutant strain, soluble homodimeric BMP6 can be formed. The overexpressed MBP-BMP6 fusion protein is purified by chromatography, and shown to be functionally active.
Bone Morphogenetic Protein 6 ; biosynthesis ; genetics ; Carrier Proteins ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Maltose-Binding Proteins ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Solubility ; Transformation, Bacterial

Objective:To investigate the effectiveness of case-based learning (CBL) combined with online teaching in standardized residency training of rheumatology and immunology.Methods:A total of 78 individuals who participated in standardized residency training in Department of Rheumatology and Immunology in our hospital from June 2019 to August 2020 were included and divided into observation group and control group. The individuals in the control group received traditional teaching, and those in the observation group received CBL combined with online teaching. The physicians receiving standardized residency training were evaluated by theoretical examination, clinical operation skill assessment, and instructor rating, and the degree of satisfaction with teaching, degree of satisfaction with teaching methods, and classroom learning atmosphere were also evaluated.Results:The observation group had a theoretical examination score of (94.10±2.01) and a clinical operation skill assessment score of (90.44±1.57), which were significantly higher than those of the control group ( P<0.05), and the observation group had a significantly better instructor rating (89.36±1.33) than the control group ( P<0.05). Compared with the control group, the observation group had significantly higher degree of satisfaction with teaching (3.79±0.41), degree of satisfaction with teaching methods (3.92±0.27), and evaluation of classroom learning atmosphere (3.90±0.31) ( P<0.05). Conclusion:CBL combined with online teaching can help to improve learning efficiency, stimulate the enthusiasm for learning, expand clinical thinking, promote the growth of teaching and learning, and form a virtuous cycle among trainees receiving standardized residency training, which holds promise for further exploration.

Objective To investigate the interventional effect and mechanism of a novel ampe-lopsis hydrogel on dampness-heat accumulation syndrome of gouty arthritis.Methods A total of 90 patients with gouty arthritis who met the diagnostic criteria of western medicine and were differentiated as damp-heat accumulation syndrome of traditional Chinese medicine(TCM)were randomly divided into treatment group,control group and blank group,with 30 patients in each group.The blank group was treated with etoricoxib only,the control group was treated with etoricoxib combined with ampelop-sis hydrogel,and the treatment group was treated with etoricoxib combined with external application of ampelopsis hydrogel.The clinical efficacy,time to symptom improvement,safety,comfort,changes in syndrome scores of TCM,serum inflammatory factors[C-reactive protein(CRP),tumor necrosis factor-α(TNF-α),erythrocyte sedimentation rate(ESR)],NF-κB signaling pathway-related pro-teins,Visual Analogue Scale(VAS)scores for pain,and joint mobility were compared among the three groups before and after treatment.Results The total effective rates in the treatment group and control group were 93.33％and 90.00％,respectively,which were higher than 70.00％in the blank group(P＜0.05).The time for improvement of pain,redness,tenderness,and limited joint mobility in the treatment group was shorter than that in the control group and blank group(P＜0.05).After 7 days of treatment,the TCM syndrome score in the treatment group was lower than that in the con-trol group and blank group,and the levels of serum CRP,TNF-α,and ESR and the expressions of NF-κB signaling pathway-related proteins P50 and P65 in the treatment group and control group were lower than those in the blank group(P＜0.05).After 7 days of treatment,the VAS score in the treatment group was lower than that in the control group and blank group,and the comfort score in the treatment group was higher than that in the control group(P＜0.05).There was no significant differ-ence in the incidence of adverse reactions among the three groups(P＞0.05).Conclusion The effect of ampelopsis hydrogel in treating gouty arthritis is better than that of ampelopsis paste,and its mechanism may be related to the regulation of the NF-κB pathway and inhibition of inflammatory fac-tor expression.The hydrogel is easy to use,hygienic,and comfortable,and is expected to become a safe,effective,and convenient external medicine for gouty arthritis.

Objective To investigate the interventional effect and mechanism of a novel ampe-lopsis hydrogel on dampness-heat accumulation syndrome of gouty arthritis.Methods A total of 90 patients with gouty arthritis who met the diagnostic criteria of western medicine and were differentiated as damp-heat accumulation syndrome of traditional Chinese medicine(TCM)were randomly divided into treatment group,control group and blank group,with 30 patients in each group.The blank group was treated with etoricoxib only,the control group was treated with etoricoxib combined with ampelop-sis hydrogel,and the treatment group was treated with etoricoxib combined with external application of ampelopsis hydrogel.The clinical efficacy,time to symptom improvement,safety,comfort,changes in syndrome scores of TCM,serum inflammatory factors[C-reactive protein(CRP),tumor necrosis factor-α(TNF-α),erythrocyte sedimentation rate(ESR)],NF-κB signaling pathway-related pro-teins,Visual Analogue Scale(VAS)scores for pain,and joint mobility were compared among the three groups before and after treatment.Results The total effective rates in the treatment group and control group were 93.33％and 90.00％,respectively,which were higher than 70.00％in the blank group(P＜0.05).The time for improvement of pain,redness,tenderness,and limited joint mobility in the treatment group was shorter than that in the control group and blank group(P＜0.05).After 7 days of treatment,the TCM syndrome score in the treatment group was lower than that in the con-trol group and blank group,and the levels of serum CRP,TNF-α,and ESR and the expressions of NF-κB signaling pathway-related proteins P50 and P65 in the treatment group and control group were lower than those in the blank group(P＜0.05).After 7 days of treatment,the VAS score in the treatment group was lower than that in the control group and blank group,and the comfort score in the treatment group was higher than that in the control group(P＜0.05).There was no significant differ-ence in the incidence of adverse reactions among the three groups(P＞0.05).Conclusion The effect of ampelopsis hydrogel in treating gouty arthritis is better than that of ampelopsis paste,and its mechanism may be related to the regulation of the NF-κB pathway and inhibition of inflammatory fac-tor expression.The hydrogel is easy to use,hygienic,and comfortable,and is expected to become a safe,effective,and convenient external medicine for gouty arthritis.

Paridis Rhizoma， a traditional valuable Chinese herbal medicine， has the functions of clearing heat and removing toxin， relieving edema and pain， cooling liver and calming convulsion， which can be used to treat various diseases such as mumps， abscess， burn， bleeding， and tumor. It has been used in folk medicine for a long time and is the main raw material of various Chinese patent medicines such as Gongxuening Capsules and Yunnan Baiyao. Polyphyllin Ⅰ， an isospirostanol saponin and one of the main active components in Paridis Rhizoma， is distributed in the rhizome， pericarp， and leaves of Paris polyphylla. With high polarity， polyphyllin Ⅰ is mainly extracted by n-butanol extraction and macroporous adsorption resin chromatography， separated by silica gel column chromatography and preparative high performance liquid chromatography， and purified with the combination of methods. With anti-tumor， anti-inflammatory， antibacterial， and anti-virus effects， it is generally employed to treat liver cancer， lung cancer， gastric cancer and other cancers as well as arthritis， influenza， sore toxin， and bacterial infection. However， polyphyllin Ⅰ may cause stomach irritation， hemolysis， liver damage， kidney damage， heart damage， and other adverse reactions. Pharmacokinetic studies show that it has problems such as low bioavailability and poor intestinal absorption and permeability， which affect the clinical application of polyphyllin Ⅰ. This paper summarizes the research on the plant sources， extraction and separation methods， pharmacological effects， adverse reactions， and pharmacokinetics of polyphyllin Ⅰ in recent years， which is expected to provide a reference for the rational clinical application and other in-depth research work of polyphyllin Ⅰ.

As a rare Chinese medicinal material， Paridis Rhizoma is mainly distributed in Yunnan， Guangxi， and Guizhou in southwestern China， with the effect of clearing heat and detoxifying， alleviating edema and relieving pain， cooling liver and tranquilizing mind. It is particularly effective for injuries from falls， fractures， contusions and strains， snake bites， cold wind-induced convulsion， and other diseases， which has been used for more than 2 000 years. According to modern research， polyphyllin Ⅱ， one of the main active components of Paridis Rhizoma， belongs to diosgenin in structure. It has the anti-tumor， anti-inflammatory， antiviral， antibacterial， immune-regulating， antioxidant， and multidrug resistance-reversing activities， showing good application prospect. Especially， the anti-tumor effect of polyphyllin Ⅱ has attracted wide attention， and the mechanism is inhibiting proliferation， migration， and invasion of tumor cells， inducing cell cycle arrest， apoptosis， and autophagy， suppressing angiogenesis， and modulating tumor microenvironment. However， the pharmacokinetic results show that polyphyllin Ⅱ has low bioavailability in vivo due to the low solubility， poor absorption， unsatisfactory distribution， and slow metabolism， which limit the clinical application. In recent years， there has been an explosion of research on the adverse reactions of polyphyllin Ⅱ， such as the strong hemolytic activity and obvious cytotoxicity to liver， kidney， myocardium and cardiovascular cells. Thus， papers were retrieved from "CNKI"， "VIP"， "Wanfang Data"， "PubMed"， "Web of Science"， and "Elsevier SD" with "Paris saponin Ⅱ"， "Polyphyllin Ⅱ" as the main keywords， and the pharmacological activities and mechanisms， pharmacokinetics， and adverse reactions were summarized. The findings are expected to serve as a reference for the in-depth research， development， and utilization of polyphyllin Ⅱ.