To measure the relative affinity and type Specificity of neutralizing monoclonal antibodies against the hexon antigen of adenovirus type 7,a simple ELISA double antibody binding system has been developed.By this method,ralative affinity of nine MAbs was estimated from the antibody concentrations at approximately 50% of plateau binding and were ranked.The theoretical basis for this method was discussed. Hybridomas secreting antibodies of desired affinity can be selected at an early stage after fussion by measuring relative affinity of hybridomas supernatants.The type specificity was judged by the difference between end-point concentration against adeno- virus type 7 and type 3.And the relation between affinity and specificity of MAbs was discussed.

Glycyrrhiza uralensis Fisch. ex DC is widely used in traditional Chinese medicine (TCM). Among its various active components, glycyrrhizic acid is believed to be the marker component. Squalene synthase (SQS) and beta-amyrin synthase (beta-AS) are key enzymes in the biosynthetic pathway of glycyrrhizic acid in G uralensis. To reveal the effects of co-expression of SQS1 and beta-AS genes on this pathway, 7 yeast expression vectors harboring different SQS1 variants and beta-AS were constructed and expressed in Saccharomyces cerevisiae as fusion proteins. TLC and GC-MS results showed that co-expression of SQS1 and beta-AS enhanced the accumulation of beta-amyrin. The effects of SQS12 were more obvious than the other two SQS1 variants. This study is significant for further investigations concerned with exploring the biosynthesis of glycyrrhizic acid in vitro and strengthening the efficacy of G. uralensis by means of increasing the content of glycyrrhizic acid.
Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Glycyrrhiza uralensis ; genetics ; Intramolecular Transferases ; metabolism ; Oleanolic Acid ; analogs & derivatives ; metabolism ; Plant Proteins ; genetics ; Recombinant Proteins ; metabolism ; Saccharomyces cerevisiae ; metabolism

Objective To explore the value of real-time tissue elastography (RTE) with tissue dispersion quantitative analysis technique for assessment of liver fibrosis stage.Methods 51 rats were injected 6％ thioacetamide to induce liver fibrosis model,and 9 rats were injected saline as control group.In modeling 4 weeks,8 weeks,12 weeks respectively,14 rats in group of liver fibrosis model and 3 rats in control group were randomly selected to RTE.All the rats underwent tissue dispersion quantitative analysis,to obtain 12 quantitative parameters of elasticity,which included average relative strain value (MEAN),standard deviation of relative strain value (SD),area ratio of low-strain region (％ AREA),complexity (COMP),kurtosis (KURT),skewness (SKEW),contrast (CONT),entropy (ENT),inverse difference moment (IDM),angular second moment (ASM),correlation (CORR) and liver elasticity index (LF index).Subsequently,rats were sacrificed and their livers were taken for pathology analysis.Liver fibrosis model group was divided into S0,S1,S2,S3,S4 group.The 12 quantitative parameters of elasticity were compared with each group.Results 49 rats were successfully modeled,and 42 rats were analyzed.Except COMP,KURT,CORR,the other quantitative parameters had statistically differences (P ＜ 0.05).The other 9 parameters were correlated with liver fibrosis stage.Among these parameters,MEAN,％ AREA and LF index had higher related coefficient(r =-0.831,0.882,0.866).The ROC curve was made by MEAN,LF index and ％AREA to estimate the fibrosis stage,when S≥S1,S≥S2,S≥S3,S =S4,the areas under the ROCcurve were 0.884,0.925,0.934,0.962 (MEAN);0.917,0.958,0.984,0.962 (％AREA);0.917,0.948,0.966,0.967 (LF index),respectively.Conclusions As a non-invasive examination,RTE dispersion quantitative analysis technology can be used to quantitatively assess liver fibrosis.

β-Amyrin synthase (β-AS) genes of Glycyrrhiza uralensis from 6 different regions were analyzed by PCR-SSCP and sequenced, then the correlationship between β-AS SNP and regions of Glycyrrhiza uralensis were determined. According to the 1 coding single nucleotide polymorphism on the first exon of β-AS gene at 94 bp site, Glycyrrhiza uralensis could be divided into 3 genotypes. In these genotypes, the percentage of 94A type in genuine regions was much higher, and it had significant differences with the percentage in non-genuine regions (P < 0.001). The results of the experiment proved that different β-AS genotypes at 94 bp site from different regions may be one of the important reasons to result in the genuineness of Glycyrrhiza uralensis.
Exons ; Genotype ; Glycyrrhiza uralensis ; classification ; enzymology ; genetics ; Intramolecular Transferases ; genetics ; Plant Proteins ; genetics ; Polymorphism, Single Nucleotide ; Polymorphism, Single-Stranded Conformational

OBJECTIVETo assess the efficacy of tendons of minimally invasive therapy (TMIT) combined drug therapy by comparing it with treatment by drug therapy alone on patients with knee osteoarthritis (KOA).

METHODSTotally 60 KOA patients were assigned to the treatment group and the control group according to random digit table, 30 in each group. Patients in the control group took Hydrochloric Acid Glucosamine Capsule and Celecoxib Capsule. Patients in the treatment group additionally received TMIT. The treatment course for all was 4 weeks. Scores for visual analogue scale (VAS) and the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index were observed and recorded at week 1 and 4 after treatment by acupotomology mirror.

RESULTSCompared with before treatment, improvement was shown in VAS score, pain and stiffness degrees, activities and functions, and WOMAC scores at week 1 and 4 after treatment in all patients with statistical difference (P < 0.05). Besides, better effect was shown in the treatment group (P < 0.05).

CONCLUSIONSTMIT combined drug therapy could relieve KOA patients' pain, stiffness and joint activities, elevate the overall efficacy. TMIT was easily operated with less injury.

Celecoxib ; Drug Therapy, Combination ; methods ; Humans ; Knee Joint ; Osteoarthritis, Knee ; drug therapy ; Pain ; Pain Measurement ; Tendons ; Treatment Outcome

Atmosphere ; chemistry ; Chromatography, High Pressure Liquid ; Polycyclic Aromatic Hydrocarbons ; analysis ; Sensitivity and Specificity

Objective To evaluate the right ventricular (RV) systolic function in patients with systemic lupus erythematosus (SLE) by speckle tracking imaging (STI).Methods A total of 40 cases with SLE were divided into two groups,SLE (24 cases) and SLE complicated with pulmonary arterial hypertension (PAH) (16 cases).Meanwhile,35 healthy subjects were used as control group.Longitudinal peak systolic strain (PSS) of the basal,mid,apical segments of the free wall of right ventricular and interventricular septum and suben-docardial myocardium of the whole right ventricular were measured by STI respectively.Every parameter was compared.And the main factors affecting the strain of the whole right ventricule were analyzed.T test,one-way ANVOA,SNK-q,Spearman's correlation and Pearson's correlation analysis were used for statistical analysis.Results ① In SLE group,strains of the whole RV(GLS RV) [(-22±4)％] and all segments of RV wall[the basal segments of free wall (bas FW) (-27±8)％ were lower than that of the control group (-26±4)％,(-33±6)％,respectively (t=-4.815,P=0.000; t=-3.619,P=0.001)].Tricuspid valve orifice peak velocity E/A and right ventricular ejection fraction were significantly different between SLE group and control group (P＜0.05).② In terms of conventional parameters and the strain of all segments of RV wall,bas FW and GLS RV were different between the SLE group [(-30±7)％,(-23.7±3.5)％,respectively] and SLE with PAH group [(-22±6)％,(-18.4±2.3)％,respectively] (t=-3.581,P=0.001; t=-5.769,P=0.000).③ The RV anterior wall thickness of the moderate-severe subgroup was significantly higher than that of the mild group [(5.0±0.8) mm,(4.3±0.8) mm; t=-2.49,P=0.017].GLS RV[(-19±4)％,(-24±7)％;t=-3.016,P=0.015],bas FW [(-22±9)％,(-35±6)％; t=-3.125,P=0.03] and bas IVS [(-19±6)％,(-24±6)％; t=-2.272,P=0.029] were significantly lower than that of the mild group.Conclusion Right ventricular function is decreased in patients with SLE.STI can be used objectively to assess the right ventricular longitudinal systolic function.

Objective To explore the feasibility of high-frequency ultrasound detecting median nerve (MN) in type 2 diabetes mellitus(T2DM)with diabetic peripheral neuropathy(DPN).Methods Forty-two healthy volunteers were included as control group(Group A).Eighty-eight T2DM patients were divided into two groups according to symptoms,signs and electrophysiological results:with DPN (group C,46 patients) and without DPN (Group B,42 patients).The width(W),thickness(T) and cross-sectional area(CSA) of MN at 6 points [the wrist crease,pisiform bone,hamate bone,the 6 cm proximal to the wrist crease,the 4 cm proximal to tip of the medial epicondyle(ME) and the midpoint of the humerus] were evaluated.ROC curve was plotted to determine clinical diagnostic value of three lines on six points in diagnosis of DPN. Results ①Comparing with Group A,the width(P <0.05) except for the 4 cm proximal to tip of ME and the midpoint of the humerus,the thickness (P <0.05) except for the 4 cm distal to ME and CSAs of all points were enlarged in DPN patients(P <0.001).The width(P <0.05)at the wrist crease,hamate bone and the midpoint of the humerus,the thickness (P <0.01) at the wrist crease and pisiform bone,CSAs (P <0.01) of all sites except for the wrist crease were increased in Group B.Comparing Group B with C, the width at the wrist crease,the thickness in the midpoint of the humerus,CSAs except for the 4 cm proximal to ME and the midpoint of the humerus were dramaticlly increased in Group C(P <0.001).②A diagnostic value comparison:when diagnostic value of W,T and CSA at six points is the most predictive, the sensitivity is 80.4%,84.8%,84.8% respectively and specificities are all 54.8%.Diagnostic value of CSA and T is superior to the width.The CSA and T of the hamate bone level were the most predictive of DPN.The corresponding optimal cut-off value was 0.097 cm 2 ,0.233 cm respectively.Conclusions The backbone and the abnormal changes of different lines of the MN in DPN can be observed by high-frequency ultrasound.The width and thickness of the MN at six points are increased in different degree,but CSAs of all levels can enlarge.When diagnostic value of W,T and CSA at six points is the most predictive for DPN, the sensitivity is superior to specificity respectively.The CSA and T of the hamate bone levelare the most predictive for the evaluation of DPN.

BACKGROUND:Most bone marrow mesenchymal stem cel s are infused intravenously and have very low efficiency of homing to the bone marrow. However, cel infusion via the femoral approach is little reported. OBJECTIVE:To explore the distribution of luciferase gene modified red fluorescent protein transgenic bone marrow mesenchymal stem cel s in vivo through different infusion routes. METHODS:Luciferase gene modified bone marrow mesenchymal stem cel s at different gradients (5×106, 1×106, 1×105, 1×104) were seeded or injected into the in vitro pore plate or free femurs to observe the fluorescence imaging and select the best concentration of cel s. Luciferase gene modified bone marrow mesenchymal stem cel s at the best cel concentration were injected into the mice via the femur and the tail vein, respectively. The distribution of fluorescence and cel number in the mice were explored by using bioluminescence, pathological examination, flow cytometry and quantitative PCR. RESULTS AND CONCLUSION:Ex vivo fluorescence intensity of luciferase gene modified bone marrow mesenchymal stem cel s was positively correlated with the cel concentration;fluorescent cel s in vivo appeared in the femur first and then quickly spread to the lungs in the femur group, while fluorescent cel s in the tail vein group spread to the lungs quickly after cel infusion. Fluorescent cel s could be seen in the spleen, liver and other organs 24 hours later in the two groups. The distribution and migration of cel s in mice could be observed successful y by bioluminescence;5 minutes after cel infusion, the lungs of mice in the two groups began to emit fluorescence that could spread to the liver, spleen and other tissues 24 hours later, and the fluorescence intensity reached its peak after 15 minutes. The distribution of bone marrow mesenchymal stem cel s in mice had no significant difference between the femur group and the tail vein group. To conclude, cel injection through the bone marrow cavity and tail vein fails to promote the homing of bone marrow mesenchymal stem cel s to the bone marrow.

Objective To analyze the sensitivity of clinical isolates of Candida albicans to flucona- zole,to detect mutations in their ERG11 genes,and to investigate the correlation between ERG11 gene mutation and resistance to fluconazole.Methods Candida albicans was identified from clinical isolates of Candida spp..The sensitivity to fluconazole was detected in vitro by microdilution-basesd method and Rosco tablets method.Three pairs of primers were designed to amplify three fragments of ERG11 gene(483 bps, from 295 bp to777 bp;482bps,from 723 bp to 1204 bp;489 bps,from 1179 bp to 1667 bp)after the extracting of genomic DNA.PCR products were sequenced.Results Eighty clinical isolates of Candida spp.were collected,which included 52 isolates of Candida albicans,all of which were sensitive to flucona- zole.Nineteen mutations were detected in ERG11 gene of 5 fluconazole-sensitive clinical isolates.Of the 19 mutations,14 were samesense mutations,and the remaing 5 missense mutations(T495A,A530C, G640A,A945C and G1609A),resulting in amino acid substitution D116E,K128T,E165K,E266D and V488I,respectively in lanosterol 14 alpha-demethylase.E165K was a novel mutation.Conclusions The clinical isolates of Candida albicans were highly sensitive to fluconazole;E165K and V488I might not lead to the resistance of Candida albicans to fluconazole.