G-protein coupled receptor (GPCR) family is the large st group of therapeutic targets. High throughput screening (HTS) plays critical ro les in early stage of drug discovery. Based on signaling pathway stimulated by l igand binding on the GPCRs, many functional assay technologies have been develop ed for HTS. These technologies include Fluorometric microvolume assay technology (FMAT), Fluorescence polarization (FP), Competitive enzyme-linked immunosorben t assay (ELISA), Scintillation proximity assay (SPA), Melanophore assay, Reporte r gene assay and Calcium assay. The principles and applic- ations of these technologies were summarized in this review. We particularly emphasize those techniques of nonradioactive, su bstrate and cofactor free assays, which will be the major approaches for HTS, su ch as reporter gene and calcium assays.

Objective To study the apoptosis of PBLC in SLE patients and the roles of caspase in the pathogeneses of SLE.Methods PBLC apoptosis was evaluated by vitro cell culture in 25 patients with SLE(11 active stage,14 catabasis),and 18 controls.The expression of caspase of PBLC was also detected by Western blot.Results In SLE,compared with controls,the apoptosis of PBLC were reduced before cell culture while those were increased after culture 24 and 48hrs.Expressions of caspase-2,3 in active stage patients were lower than those of catabasis group and controls.Conclusions Abnormal PBLC apoptosis results in decreasing Caspase2,3 in SLE patients.

Objective Three kinds of different internal promoters were inserted into the lentiviral vector to drive the expression of CD19-specifc chimeric antigen receptor (CAR)-redirected T lymphocytes with the green fluorescent protein(GFP) and the GFP expression efficiency of transduced cells driven by lentivirus were compared.Methods Three GFP reporter lentivirus vectors carrying different promoters and anti-CD19-CAR,including CMV,EF1α,and CMV-EF1α were selected.Three different vectors' names abbreviate to pHAGE-CMV-EFlα-EGFP-CD19,pHAGE-CMV-EGFP-CD19,pHAGE-EF1α-EGFP-CD19.Human embroic kideny 293T cells were cotransfected with the three plasmids by calcium phosphate DNA precipation.And the expression of GFP was observed under fluorescent microscope after transfecting 293T cells,and virus supernatant was collected after 72 h and centrifuged.Nucleic acid copy number in RNA and the expression of EGFP and CD3 zeta were identified through fluorescence quantitative PCR,flow cytometry and western blotting,respectively.The titers of the lentiviral vectors were determinde by scoring GFP expression following swerial dilutions of the viral supernatant on 293T cells.T lymphocytes was infected with lentivims produced from these reporter vectors.Then,fluorescence microscope,wesyern blotting were used to detect the GFP expression strength.Results Results of fluorescence microscope maintained that different internal promotors driving the expressing effect is different.293T as targeted cell,the transfected 293T cells were found containing strong expression of GFP.The amount of the 293T cells expressing GFP driven by CMV-EF1α promoter was the largest,while the EF1α promoter was the smallest.Flow cytometry results show that the rate of transduction is 72.4％,20.6％ and 14.5％,respectively.T lymphocytes as targeted cell,Western blot showed that CMV-EF1α was the largest among all promoters,while the CMV was the smallest.Conclusion In the study of gene expression mediated by lenuvirus,proper cell lines and promoters should be selected to obtain high efficiency.

Objective To investigate the effect and mechanism of p38 MAPK inhibition in reducing the damage to rat's hippocampal neurons caused by kainic acid (KA) and observe the three-dimensional morphological changes on the cell surface. Methods The rat's hippocampal neurons cultured primarily for 10 days were pretreated with SB203580 (0.2 ?mol/L, a p38 MAPK inhibitor). Thirty minutes later, the hippocampal neurons were administered with KA at concentrations of 0, 25 and 250 ?mol/L for action for 10 and 100 minutes respectively. The cellular membrane structure was scanned and examined at nano-level by using atomic force microscope. Results Normal neurons displayed smooth membrane surface with homogeneous and regular undulation. In contrast, the neurons treated with KA showed coarse membrane surface with holes. Furthermore, the degree changes increased with the action time and the KA concentrations in a dose-effect dependent fashion. The above-mentioned changes were obviously mitigated by means of pretreatment with SB203580 (200 nmol/L). Conclusions Inhibition of p38 MAPK may, in certain degrees, protect the neurons against the impairments on cytomembrane resulted from the toxic effect of KA.

Aplastic anemia (AA) refers to a life-threatening bone marrow failure disorder.With respect to the precise pathophysiology of AA at present, it is still unclear.MicroRNA (miRNA), about 22 nucleotides in length, is a kind of small RNAs and it can regulate gene expression post-transcriptionally.According to domestic and foreign reports recently, there are abnormal expressions of several miRNAs in AA patients, suggesting that miRNAs may be involved in the development and progression of AA.This article reviews the study progress of miRNAs in AA.

Objective To confirm the role of human parvovirus B 19 ( B19) infection in childhood immune thrombocytopenia ( ITP) patients.Methods A total of 416 cases of newly diagnosed childhood ITP patients from Jan .2011 to Dec.2013 had been summarized to be cases group , Then a total of 130 childhood patients with common respiratory tract infection were selected randomly as the control group . All patients had been divided in grouped by age as <1 years group(n=187), 1-3 years group(n=127), 3-7 years group(n=71), and 7-14 years group(n=31).The incidence rate of B19 infection in all age groups, the prognosis conditions in B19 infection positive and negative groups after the same therapy protocol were obserred .Results The B19 infection rate had no statistical significance in re-search groups and control groups .And no significant difference in the same age groups compared with control groups (all P>0.05) was found.All the ITP patients had not been given anti -B19 treatment.The PLT remission rate,respectively, after treated with the same pro-tocol including glucocorticoid and/or immunoglobulin had a declining trend as the ages increasing .The B19 infection groups of all ages al-so had no significant difference PLT remission rate had been confirmed in non -B19 infection patients in each age groups (all P>0.05). Conclusions B19 infection may not be a major cause in childhood newly ITP , and treatment protocol with no anti -B19 treatment had no influence on the clinical curative efficacy .

Objective] The pathogenesis of HuangQin Tang is discussed little in six meridians syndrome differentiation, and which meridian syndrome does it belong to isn’t clear. So this article tries to discuss the belonging and pathogenesis of it. And the key point of using it in clinic will also be involved.[Method] Combining the observation of efficacy of XiaoChaiHu Tang and HuangQin Tang and the elaboration articles are related to that two prescriptions to analyze the difference of that two prescriptions in pathogenesis.[Result]It’s found that HuangQin Tang and XiaoChaiHu Tang all belong to ShaoYin in six meridian syndrome.But their pathogenesis is different. HuangQin Tang should be used in heat syndrome of ShaoYang disease which caused by heat pathogen of ShaoYang meridian and diarrhea will appear because belly is shocked. However, XiaoChaiHu Tang is used to drive cold pathogen of ShaoYang meridian. [Conclusion]Both of HuangQin Tang and XiaoChaiHu Tang belong to ShaoYang Meridian, but it’s known that HuangQin Tang is used less. And less discussion of HuangQin Tang should be blamed. The more we know on pathogenesis of HuangQin Tang, the better we use it.

Objective To investigate the effect of HPA gene polymorphism and its alleles on platelet transfusion.Methods From January 2015 to January 2017,50 patients suffered from Platelet transfusion refractoriness were collected as an observation group while 50 patients without Platelet transfusion refractoriness were collected as a control group.The HPA gene polymorphism of the both groups were compared.Results When compared with the control group,the HPA type coincidence rate of Donor and recipient in the observation group decreased significantly (4.00％ vs.82.00％,P =0.000).And patients in the observation group got decreased levels of percent platelet recovery and platelet corrected growth index.(HPA16,9,15) were common in both groups.However,aa,ab and bb of the HPA-2 allele got 37,8 and 5 cases in the observation group,while there were 48,2 and 0 cases in the control group (P=0.007).And aa,ab and bb of the HPA-3 allele got 33,9 and 8 cases in the observation group,while there were 46,3 and 1 cases in the control group (P =0.005).Moreover,aa,ab and bb of the HPA-15 allele got 35,8 and 7 cases in the observation group,while there were 47,2 and 1 cases in the control group (P =0.007).Homozygous aa were the most common gene type in the donors of the both groups,and the difference was no statistically significant (P＞0.05).As aa,ab and bb of the HPA-2,3 change,the percent platelet recovery and platelet corrected growth index both decreased (P＜0.05).Conclusion Platelet transfusion refractoriness is related to HPA gene polymorphism,and allele b may be the cause of platelet transfusion refractoriness.

Objective To establish a method for accurate typing of ABO in blood donors and patients with difficult blood types,and to provide clinical reference for safe blood transfusion,organ transplantation,and bone marrow matching.Methods The results of serological and genotyping of 178 volunteers from four ethnic groups in China were analyzed statistically,and the blood type difference coincidence test was carried out.Results The difference of blood type difference between the four nationalities was checked (x2 =24.5,P＞0.05).The results showed that there was no difference in blood group distribution between the four nationalities.Conclusion Serological results of 178 volunteers from four ethnic groups in China are consistent with the results of molecular biology,and there is no difference in blood type distribution.

Objective To explore the new method to prevent graft versus host disease (GVHD) by clearing T lymphocytes of bone marrow graft through Fas-FasL way. Methods FasL-cDNA was transfected into BALB/C mouse hematopoietic cells by liposomes. The transfected cells were cultured together with BAC mice bone marrow graft. The mixed bone marrow graft was injected into BALB/C mouse recipients after 60 Co-? ray irradiating. Then the mortality, manifestation and pathologic change of GVHD of recipient mice were observed. Results Compared with control group, the mortality of the recipients in the experimental group was markedly decreased in 60 days (20% vs 70%, P