Objective To evaluate the diagnostic value of serum albumin (Alb),total protein (TP),body mass index (BMI),Mini Nutritional Assessment (MNA) and hand strength in the nutritional status in maintenance hemodialysis (MHD) patients.Methods A total of 126 MHD patients were included in this study who had been on MHD for at least 3 months.Depending on the levels of Alb,patients were divided into two groups:normal nutrition group (group A) and malnutrition group (group B).TP,BMI,MNA and hand strength were also detected at the same time.Independent samples t test,Spearman correlation analysis,ROC curve were used to analyze their difference between the two groups and evaluate their diagnostic value in nutritional status in MHD patients.Results Age,sex,height,weight and dialysis ages had no statistical significant difference (P ＞ 0.05) between group A and group B,while Alb,TP,BMI,MNA and hand strength had statistical significant difference (P ＜ 0.05) between two groups.After adjusting for age,sex and hemodialysis age,Alb,TP,BMI,MNA and hand strength were positively correlated with each other (P ＜ 0.05).Since the area under the ROC curve of BMI was the smallest,BMI had the lowest diagnostic value in evaluation of the nutritional status in those patients.Conclusions Alb,TP,MNA and hand strength are good indexes in evaluation of the nutritional status in MHD patients but BMI is not.

Objective To establish a flow cytometry-based drug susceptibility test for the rapid de-tection of antifungal susceptibility or resistance of Candida isolates.Methods The gate selection and opti-mal experimental conditions of flow cytometry-based drug susceptibility test were determined by using Candi-da albicans strain ATCC90029 as the test strain and propidium iodide ( PI) as the fluorescent dye .The es-tablished flow cytometry-based drug susceptibility test was used to detect the susceptibility or resistance to fluconazole or voriconazole of 110 isolates belonging to Candida species, and the obtained results were com-pared with those by using typical M 27-A3 constant dilution method .Results The killed and viable Candida albicans ATCC90029 strains were clearly divided into two groups on the figure of SS /log (FL3) by regulating voltages.There was a high correlation between the results of susceptibility test and the proportions of killed and viable fungi in mixture (r=0.999).The flow cytometry-based drug susceptibility test could provide the results within 30 min and its optimal concentration of fungal suspension , time of drug-fungus incubation , dyeing method and time were 1.0×106/ml, 3 h incubation and sodium deoxycholate-pretreated plus PI dye-ing for 5 min, respectively .The total coincident rates between the established test and the constant dilution method were 98.2%and 87.3%in the detection of drug susceptibility of 110 fungal isolates to fluconazole and voriconazole .Conclusion The flow cytometry-based drug susceptibility test shows advantages of rapidi-ty, accuracy and high sensitivity compared with the constant dilution method .It has a great potential for clin-ical application .

Objective:To interpret the main revision about the test method for bacteriostat effect in Chinese Pharmacopoeia(2015 edition). Methods:The main difference of the bacteriostat effect test method in Chinese Pharmacopoeia(2015 edition) and (2010 edi-tion) was compared. Results:The bacteriostat effect test method in the 2015 edition was revised at a comparatively large scale in the positioning of bacteriostat effect test, product classification, assessment criteria and so on. Conclusion: The bacteriostat effect test method in Chinese Pharmacopoeia (2015 edition) gradually improves the check standards in line with the international standards.

Profilin-Ⅰis a small actin monomer-binding protein involved in regulating actin polymerization.It plays an important role in cell proliferation,differention,motility and signals transduction in different cell types including endothelial cells and vascular smooth muscle cells.This article recapitulates the wealth of information on structure,function of profilin-Ⅰand its potential role in the genesis and development of cardiovascular diseases.

Objective To screen and identify the predominant T-and B-cell ( T-B) combined an-tigenic epitopes on the outer membrane protein Loa22 of pathogenic Leptospira interrogans ( L.interrogans) stains and to further analyze their immunogenicity.Methods PCR analysis was used to detect loa22 gene in L.interrogans strains belonging to eight different serogroups or serovars prevalent in China.The PCR prod-ucts were sequenced after T-A cloning.A prokaryotic expression system for loa22 gene of L.interrogans sero-group Icterohaemorrhagiae serovar Lai strain Lai was constructed.The expressed recombinant protein rLoa22 was extracted by Ni-NTA affinity chromatography.The rabbit anti-rLoa22 serum samples and IgG were pre-pared.The T-B combined antigenic epitopes on Loa22 protein were predicted by using professional bioinfor-matic softwares.Phage display in combination with Western blot assay and ELISA were performed to identify the immunogenicity of the recombinant phage PⅢ protein-displayed and artificially-synthesized T-B com-bined antigenic epitopes, respectively.MTS assay and ELISA were performed to detect the activation of T cells and the expression of IL-2, IL-4 and IFN-γinduced by T-B combined antigenic epitopes, respectively. Results All of the tested pathogenic Leptospira strains were positive for loa22 gene, sharing 85.5%-99.8%homologies in nucleotide sequences and 93.9%-99.5%homologies in amino acid sequences.The construc-ted prokaryotic expression system for loa22 gene efficiently expressed the rLoa22 protein.Among four T-B combined antigenic epitopes (Loa22-77, Loa22-90, Loa22-125 and Loa22-157), only Loa22-90 epitope presented a strong positive band in Western blot analysis.The proliferation of CD4+T cells and the expres-sion of IL-2 ( Th1 ) and IL-4 ( Th2 ) were significantly enhanced by the stimulation with Loa22-90 epitope peptide (P<0.05).Conclusion Loa22 protein is a sequence-conserved genus-specific outer membrane protein of L.interrogans.The Loa22-90 epitope is the predominant T-B combined antigenic epitope of Loa22 protein, which might be used as a candidate antigenic epitope in the development of multiple antigenic pep-tide ( MAP) vaccines against Leptospira infection.

Objective To establish an animal model of metabolic syndrome which is prone to atherosclerosis. Methods Eight rabbits were randomly selected from in total of 16 rabbits as control group who were fed with normal rabbit feed. And the rest were used as model group, who were fed with high fat diet. Animals were weigh every week, and triacylg-lycerol (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), and glucose (GLU) content were tested in the 8th, 10th , 13th, 16th and 18th week. Femoral arterial blood pressure and insu-lin (INS) content were assessed at the end of experimental period which is the 18th week;Also, aortic sclerosis rating, arterial stiffness index, abdominal fat coefficient, liver coefficient and cardiac coefficient were calculated and heart, aorta, liver and lower extremity artery pathology were examed. Results Compared with the control group, weight in the model group in-creased significantly;Serum levels of TC, HDL-C, blood pressure abdominal fat coefficient, liver coefficient were significant-ly increased in model group;the whole blood viscosity, plasma viscosity and blood pressure fat coefficient, liver coefficient, aortic sclerosis rating, arterial stiffness index, and GLU all increased significantly in model group;while HDL-C/TCHO, in-sulin sensitivity index decreased significantly in model group(P < 0.05 or P<0.01). Conclusion Food induced method can form metabolic syndrome and atherosclerosis in rabbit model.

Objective:To interpret the main revision of the microbiological examination for nonsterile products:tests for specified microorganisms in Chinese Pharmacopoeia (2015 edition). Methods:The microbiological examination for nonsterile products:tests for specified microorganisms in Chinese Pharmacopoeia (2015 edition) was compared with the relevant content in the 2010 edition, and then the differences were investigated. Results:Microbiological examination for nonsterile products:tests for specified microorganisms in Chinese Pharmacopoeia (2015 edition) had been revised at a comparatively large scale in the inspection items, test method, micro-bial culture system, the quality control concept and so on. Conclusion:Microbial inspection system in Chinese Pharmacopoeia (2015 edition) is gradually improved to become a high standard check system in line with the international standards.

Objective To investigate the relationship of cellular immunity of the hand-foot-mouth disease (HFMD) children and the disease severity and the variation following the recovery of disease.Methods A total of 560 HFMD cases was collected,and divided into severe and common groups.Another 120 cases were collected for comparison.T cell subsets (CD3 +,CD4 +,and CD8 +) rates were tested.The difference in cell immunity in each group were compared,and the comparison of cell immunity improv-ment during acute and recovery periods was conducted at the same time.Results In the 560 cases of children with HFMD,CoxA16-positive rate in common group was higher than that in severe group (x2 =280.72,P ＜0.01,severe cases); EV71 and other virus positive rates in severe group were higher than that in common group (x2 =127.75,P ＜ 0.01,x2 =5.43,P ＜ 0.05).Cell immunity was compared among3 groups (t =9.82,4.98,3.06); CD3+,CD4+,CD8+ results,tested within 2h after admission and after 1 week,were compared between severe and common groups (common group t =7.73,3.86,4.71; severe group t =6.13,2.60,3.36).Compared to severe group,cell immunity improvement was more obvious between before and after 1-week treatment in common group (t =2.57,2.51,2.95).The difference was statistically significant (P ＜ 0.05).Conclusions According to the etiology test of children with HFMD,CoxA16-positive rate was higher in common group; EV71 and other virus positive rates were higher in severe group.Cell immunity function decreased in severe and common group at the beginning of the disease; it was,however,significantly restored after 1-week treatment; and it was related to the severity of clinical symptoms.

Objective To analyze the platelet activating factor acetylhydrolase ( PAF-AH) activity of a gene product encoded by LA2144 gene of Leptospira interrogans ( L. interrogans) , to investigate the ex-pression and secretion of LA2144 protein in various cell cultures and to further understand its function in in-ducing internal hemorrhage in an animal model. Methods The DNA sample containing LA2144 gene was extracted from L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai and used as the template for gene cloning by PCR. The LA2144 gene without the signal sequence coding region was amplified by PCR and inserted into a prokaryotic expression construct for the protein expression. The expressed recombinant protein, rLep-PAF-AH, was purified by Ni-NTA affinity chromatography. Spectrophotometry was used to measure the hydrolytic activity, hydrolytic efficiency, Km and Kcat values of the rLep-PAF-AH protein in hydrolyzing PAF substrate. Real-time fluorescent quantitative RT-PCR ( qRT-PCR) and Western blot assay were performed to measure the expression of LA2144 gene at mRNA and protein levels in human umbilical vein endothelial cells (HUVEC), human monocytes (THP-1) and murine macrophages (J774A. 1) with L. interrogans strain Lai infection, respectively. Each syrian hamster was intravenously injected with 100 μg of LPS-free rLep-PAF-AH for two times. Hemorrhage in the lungs, livers and kidneys were observed in three days after the injection. Results The constructed prokaryotic expression system for LA2144 gene of L. inter-rogans strain Lai could highly express the rLep-PAF-AH upon the induction of IPTG. The purified rLep-PAF-AH showed high purity with a single protein band in gel as indicated by SDS-PAGE. The efficiency of 5 μg of rLep-PAF-AH in hydrolyzing PAF substrate was 26. 6 U/L with a Km value of 82. 79 μmol/L and a Kcat value of 0. 24 S-1 . The expression of Lep-PAF-AH at mRNA level in HUVEC, THP-1 and J774A. 1 cells were significantly elevated after co-culture with L. interrogans strain Lai for 1 or 2 hours (P<0. 05). A large amount of Lep-PAF-AH were detected in the supernatants from co-cultures of L. interrogans strain Lai with the three cell lines, but not from the culture of the spirochete in EMJH medium. The signs of hemor-rhage were observed in the lung of hamsters injected with rLep-PAF-AH, but not in tissue samples from liver and kidney. Conclusion The LA2144 gene product was characterized by a stronger PAF-AH activity. The expression of LA2144 gene at mRNA and protein levels in various cell lines were enhanced during L. interro-gans infection. Moreover, the rLep-PAF-AH could induce the pulmonary hemorrhage in hamsters. This stud-y indicated that the protein encoded by LA2144 gene was an important virulence factor causing hemorrhage in hosts during L. interrogans infection.

Aim TostudytheroleofASIC1aonthe matrix turnover and MAPK expression of the rat articu-lar chondrocytes with extracellular acidosis. Methods ArticularchondrocyteswereisolatedfromSprague-Dawley rats, and their phenotype was determined by toluidine blue and immunocytochemical staining. The GAG content of cell culture supernatant was deter-mined by dimethyl-methylene blue spectrophotometric assay, while Hyp content by chloramine T assay. ELISA assay was used to measure MMP-2 , TIMP-2 content. Furthermore, the ERK1/2, p38 MAPK phos-phorylation protein expression levels were tested by Westernblotassay.Results ASIC1acontributedto the effect of GAG, Hyp and TIMP-2 levels reduction induced by extracellular acidification, while the effect of MMP-2 was weaker. Moreover, ASIC1a could in-crease the ERK1/2 , p38 MAPK phosphorylation pro-teinexpressionlevels.Conclusion ASIC1acould regulate rat articular chondrocytes matrix turnover via ERK1/2 and p38 MAPK signaling pathway, and there-by inhibit the rat articular cartilage damage induced by acidosis.