Firstly discovered in 1980s,human immunodeficiency virus (HIV) continues to affect more and more people.However,there is no effective drug available for the therapy of HIV infection.Betulinic acid existing in various medicinal herbs and fruits exhibits multiple biological effects,especially its outstanding anti-HIV activity,which has drawn the attentions of many pharmacists.Among the derivatives of betulinic acid,some compounds exhibited inhibitory activities at the nanomolar concentration,and have entered phase Ⅱ clinical trials.This paper summarizes the current investigations on the anti-HIV activity of betulinic acid analogues,and provides valuable data for subsequent researches.

There are few articles or reports collecting evidence about parenterally administered salvianolate from premarketing and postmarketing research or studies systematically. This article is an exact miniature of a systematical report about parenterally administered salvianolate. We analyzed information from four aspects, such as quality control reports, non-clinical premarketing safety experiments, postmarketing research (efficacy studies, hospital information system data and national spontaneous reporting system data) and literature analysis. All the four aspects build an evidence body for Kudiezi Solution in order to inform its safety use in clinical practice and further study.
Hospital Information Systems ; Humans ; Plant Extracts ; administration & dosage ; adverse effects

OBJECTIVE To observe the effect of electrolyzed oxidizing water(EOW) on the surface sterilization of indoor environment.METHODS Sterilized the surface of ground,wall,table,mob and thermostat-controlled water-bath in cell culture room and good laboratory practice(GLP) with EOW,then to draw the materials and culture with culture dish for 48 h in a 37℃ incubator,and count the colony number.Sterilization method and grouping: group A treated as a control,group B sterilized with EOW,group C sterilized with ultraviolate ray for 30 min and group D first treated with ultraviolate ray for 30 min,then sterilized with EOW.RESULTS In group A,the bacteria were overgrew and formed flakiness in 10/10 culture dishes;1 colony was formed in group B,the sterilization effective rate was 90%.The bacteria culture of group C found no bacteria growing after sterilization with ultraviolate ray,however,sample from surface and culture after sterilization were seen bacteria,though the number of bacteria was less than group A.The bacteria culture outcome of group D was negative.CONCLUSIONS The EOW has a good sterilization effect,it is safe and untoxic,costly cheap and convenient to use,and fit to claim of environmental protection.

Objective To investigate the adverse effects of taurine on rabbit corneal endothelial cells. Methods Six rabbits (12 eyes) were selected, and 6 histologic sections were prepared from each of the eyes. Rabbit corneal endothelial cells were cultured by explant culture method. Cells were innoculated on a 12-well tissue culture plate, 2%, 4%, 6%, 8% and 10% taurine solutions were added respectively (cells from the right and left eyes of the same rabbit were added the same concentration of taurine solution), and blank control was established. The growth of corneal endothelial cells was observed by inverted microscopy, and cell morphology on the 1st, 2nd, 4th, 6th and 8th day of culture was observed with Wright staining. Results Corneal endothelial cells cultured with 2%, 4% and 6% taurine solutions and those of blank control formed endothelial cell layers after culture for one week, and the cells exhibited hexagonal or round-like morphology. Corneal endothelial cells cultured with 8% taurine solution appeared to be undergrowth with small cell body on the 4th day, and cell death occurred on the 8th day. Corneal endothelial cells cultured with 10% taurine solution turned out to be undergrowth with small cell body on the 2nd day, and cell death had occurred. The same growth velocity and cell morphology were observed in the corneal endothelial cells from the right and left eyes of the same rabbit. Conclusion Taurine with concentration between 2% and 6% has no adverse effects on the growth of rabbit corneal endothelial cells.

Objective To investigate the correlation between de nove anti-endothelial cell antibodies (AECA) and adverse events after renal transplantation and the effect of transplanted renal function within six months after operation.Methods The expression of AECA in serum of 85 renal transplant recipients was detected by indirect immunofluorescence assay (ⅡF) before and 1 day,3 days,7 days,15 days and 1 month after renal transplantation.The patients were divided into double positive group (AECA positive before and after surgery),single positive group (postoperative de nove AECA positive) and negative group (preoperative and postoperative AECA negative) according to AECA test results.The occurrence of adverse events in each group such as DGF,rejection,infection and so on,as well as the serum creatinine levels were recorded at each time point within six months.Results Of the 85 recipients,29 were positive for AECA,including 19 in the double positive group,10 in the single positive group,and 56 in the negative group.The incidence of rejection in single positive group (30％) was higher than that in the rest two groups (5.3％ for double positive group,and 17.9％ for negative group),but there was no statistically significant difference (P =0.21).The incidence of DGF in the single positive group,double positive group and negative group was 70.0％,26.3 ％ and 32.1 ％ respectively.The positive rate of the single positive group was significantly higher than that of the rest two groups (P =0.04),and the duration of DGF was significantly longer in the single positive group than that of the rest two groups (P＜0.01).The incidence of infection in the single positive group,double positive group and negative group was 20.0％,21.1％ and 8.9％respectively,and there was no significant difference among the three groups (P =0.31).As compared with the double positive group and the negative group,the serum creatinine level was significantly increased in the single positive group at 1st week,1st month,3rd month and 6th month after operation (P =0.02,P =0.04,P =0.04 and P =0.02 respectively).Conclusion Postoperative AECA can increase the risk of DGF,prolong the duration of DGF,and affect the recovery of renal function within 6 months after renal transplantation.

Objective To investigate whether the donor's glomerular filtration rate (GFR) to recipient's weight ratio (Dg/Rw) is a useful tool to predict early clinical outcome in living-related do-nor transplantation.Methods A total number of 108 living donor transplant recipients in the Chinese Military 309th Hospital from Jan.2014 to July 2015 were enrolled in this study.The patients who had multi-organ transplantation or developed grafts rejection,delayed graft function,hydronephrosis or renal vascular stenosis were excluded.The 90 qualified recipients were divided into G1 group (Dg/Rw ≤0.81),G2 group (Dg/Rw 0.81～1.11),and G3 group (Dg/Rw≥1.12).We respectively analyzed the relationship between recipient's serum creatinine Scr and Dg/Rw at 3-,7-,30-day and 1 year after transplantation.Results Scr at 3-,7-,30-day and 1 year after transplantation had linear correlation with Dg/Rw.As compared with G1 and G2 groups,Scr level was significantly reduced in G3 group at different time points (P＜0.05).Conclusion Dg/Rw has a negative relationship with Scr level after renal transplantation.Pre-transplant Dg/Rw is a potential index to predict the early clinical outcome in living-related donor transplantation.

Classical swine fever virus (CSFV), an enveloped positive-stranded RNA virus in the genus Pestivirus of the Flaviviridae family, is the causative agent of a highly contagious swine disease characterized by symptoms of hemorrhagic fever and immune depression, usually leading to substantial economic losses. The serological methods for detection of CSFV antibody such as ELISA are important means for the diagnosis of CSFV and immune surveillance. It is difficult to obtain CSFV antigen with high quality using traditional method because its titration titer is low in cell culture. CSFV has four structural protein named C, E0, El and E2. The E2 protein contains major antigenic determinants that are conserved between different CSFV strains and involved in neutralization by antibodies. So recombinant E2 protein can be developed as an alternative to the intact viral antigen. So far, CSFV E2 have not been expressed in E. coli with high level. Many factors, such as the secondary structure, the stability of 5' and 3' terminus of gene, the location of SD sequence and the bias of codes, are involved in the expressing level of foreign gene in E. coli . In this study, two sites of the E2 gene sequence were confirmed to be detrimental to its expression efficiency in E. coli through the computer-aided analysis. So they were mutated using recombinant PCR without changing the amino acids sequence of CSFV E2 gene. A plasmid was constructed by inserting the mutated E2 gene into the prokaryotic expression vector pET-28a(+) and named pETE2. The E. coli competent host BL21 (DE3)lysS transformed with pETE2 could express the E2 gene at high level, amounting to 28% of the total protein of the induced recombinant bacteria at the presence of IPTG. Except the hydrophobic transmembrane domain at C terminus, the recombinant E2 protein includes the total aa sequence. So it contains all the potential linear antigen epitopes of E2 protein because hydrophobic aa region can not form epitope. The recombinant E2 protein was CSFV-specific as proved by Western blotting and indirect ELISA. The rabbits immunized with the recombinant E2 can be protect from the challenge of hog cholera lapinized virus. This is the first report that E2 gene is expressed with high level expression in E. coli. In conclusion, it is an effective measure that mutate the CSFV E2 gene to increase its expression level in E. coli. The recombinant CSFV E2 protein possess fine immunonicity and can be used the antigen for the detection of CSFV antibody.
Antigens, Viral ; genetics ; immunology ; metabolism ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Mutagenesis, Site-Directed ; methods ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Viral Envelope Proteins ; genetics ; immunology ; metabolism

Objective To study the efficacy and safety of T-614 in treating rheumatoid arthritis(RA). Methods Two hundred and eighty patients with active RA were randomly allocated to 3 groups:T-614 50 mg each day,25 mg each day or placebo.Clinical and laboratory parameters were analyzed at baseline,2,4,6,12, 18 and 24 weeks.Results The ACR response rate was significantly higher in the T-614 treatment group com- pared with the placebo group during the first 6 weeks.After 24 weeks,25 mg/d,50 mg/d dosage group and the placebo group showed 39.1%,61.3% and 24.2% in ACR20,23.9%,31.2% and 7.4% in ACR50 respectively.A time-response in ACR response after 24 weeks was observed,with clear superiority of the 25 mg/d and 50 mg/d dosage groups compared to the placebo,and 50 mg/d dosage group compared to 25 mg/d dosage group(P

Scm in volume(42/60),multiple site involvement(44/60),blood type"O"(31/41),in comparison with those of survival group,and the difference was statistically significant.C-erbB-2,p16,p53,P-gp,CD_(44) and CD_(25)expression were not significantly different in these two groups. Conclusion The clinical stage, lymph node metastasis,lymphatic tumor emboli and/or neural involvement,infiltration depth,histological dif- ferentiation,tumor volume,involvement extension are important prognostic factors in patients with gastric can- cer,while the significance of cancer-related gene expression in gastric carcinomas needs to be studied further.

Transcriptions are regulated by transcription factors. Natural transcription factors usually consist of at least two functional domains: a DNA-binding domain and an effector domain. According to this, novel artificial transcription factors are designed to up or down regulate transcription and expression of a target gene. The Cys2-His2 zinc finger domain is a DNA-binding module that has been widely used as the DNA-binding domain in artificial transcription factors. Each zinc finger domain, which comprises about 30 amino acids that adopt a compact structure by chelating a zinc ion, typically functions by binding 3 base pairs of DNA sequence. Several zinc fingers linked together would bind proportionally longer DNA sequences. According to the "bipartite complementary" library strategy, a pair of zinc finger phage display libraries were constructed. After construction of the libraries, a 9bp sequence (5'-GCAGAGGCC-3') on the promoter of SV40 was chosen as a target for next step. After parallel selection, PCR amplification, desired fragments recovery, re-ligation, and additional rounds of selection, phage enzyme-linked ELISA experiments were performed to identify specific binding clones displaying the zinc fingers with predetermined sequence-specificity to our target sequence. Then two clones with strong ELISA signals were chosen to be tested for binding both to its full target site (5'-GCAGAGGCC-3') and to sites containing single transition mutations. The binding specificity of one of the two clones (clone 3) was shown to be fairly good. The three-finger DNA-binding domain targeted to SV40 promoter, that is, zinc finger sequences on clone 3, was fused to KOX1 suppression domain KRAB and cloned into pcDNA3.1 (+) (which expression product was artificial transcription factor). The zinc fingers (which expression product was the DNA-binding domain of artificial transcription factor) and KRAB domain only (which expression product was effector domain of artificial transcription factor) were also cloned separately into the same expression vector. All constructs contained an N-terminal nuclear localization signal. Every of the vectors (including pcDNA3.1 (+) without inserting sequences) were cotransfected with pGL3-Control and pRL-TK and the activity of luciferase was used to indicate the function of product from transfected expression vectors. Our artificial transcription factor was proved to repress the expression of reporter gene efficiently,while with only DNA-binding domain or effector domain the repression was not remarkable. By adding different effector domains and changing the DNA-binding domain, artificial transcription factor would have a wide range of potential applications.
Enzyme-Linked Immunosorbent Assay ; Genes, Synthetic ; genetics ; physiology ; Models, Theoretical ; Peptide Library ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics ; Transcription Factors ; chemical synthesis ; chemistry ; metabolism ; Zinc Fingers ; genetics ; physiology