Objective To evaluate the prospects of recombinant 38000 protein of Mycobacterium tuberculosis in tuberculosis epidemic investigation and subunit vaccine preparation.Methods Physicochemical characteristics of recombinant 38000 protein was detected by P I, peptide-mapping analysis and circular dichroism,guinea pig skin test,MTT stain,and peripheral blood macrophage phagocytosis were used to investigate the roles of recombinant 38000 protein in the cell immunity.Results Recombinant 38000 protein was acidic protein,its P I, was 4 67.The number of alkaline amino acid correspond with theoretic number;The secondary structure of recombinant 38000 protein was composed of ?-helix(32 6%),?-turn(31 6%) and random coil(35 8%) Recombinant 38000 protein could induce DTH in guinea pig sensitized by Mycobacterium tuberculosis Recombinant 38000 protein enhanced phagocytosis of macrophage in mice . PBMC from 30 8% healthy donors and 25% tuberculosis patients were stimulated by the recombinant 38000 protein.Conclusion Recombinant 38000 protein may be used as diagnostic reagent and as an candidate in development of subunit vaccine.

Objective To compare expression of extracellular proteins of virulent H37Rv and attenuated H37Ra in order to search differential proteins,to provide a train of thought for studing M.TB toxicity further.Methods Extracellular proteins were extracted from H37Rv and H37Ra which were inoculated and cultured on Suton's medium for three weeks.The first dimensional electrophoresis was performed on immobilized pH gradient rod gels(pH 3～10).Then the proteins in the rod gels were separated using SDS-PAGE gels.The silver-stained gels were dried and scanned with image scanner.The 2D image analysis was performed with image Master 2D Elite 3 10.Results The most protein spots deriving from extracellular proteins of H37Rv and H37Ra strains were in acidic range.In the basic range(pI more than 9 0),the number of protein spots belong to extracellular proteins of H37Rv and H37Ra was few.Three protein spots belong to low molecular range in H37Rv strain.However,absent in H37Ra strain.Conclusions Two-dimensional gel electrophoresis is useful to separate protein in Mycobacterium tuberculosis.

Objective To investingate the role of inflammation in coronary atherosclerosis and the relationship between the inflammatory factors and coronary risk score. Methods 56 patients with acute coronary syndrome were studied, among them 26 cases were diagnosed as acute myocardial infarction (AMI) and 30 unstable angina pectoris (UAP). The study group was compared with a control group of 30 cases who were identified as normal by coronary angiography. The concentrations of serum sICAM-1 and hsCRP were determined by ELISA assay. The coronary risk score was recorded in patients with UAP. Results Serum sICAM-1 levels were significantly elevated in patients with AMI or UAP compared with that of control group, while higher hsCRP level was observed only in the patients with AMI compared with those with UAP and the control group. By linear regression analysis, only serum sICAM-1 levels were correlated with coronary risk score (r=0.445, P

Aim To explore the influence of atorvastatin on the ultramicrostructure of the membrane surface of the rabbit endothelial cells in rabbit atherosclerosis(AS)in the nanometer level.Methods A total of 44 male New Zealand white rabbits were randomly divided into 3 groups:control group consisting of 12 rabbits,AS group consisting of 16 rabbits and atorvastatin group consisting of 32 rabbits.By the end of 2nd,6th week 6~8 rabbits of each group were sacrificed and the middle segments of thoracic aortas were obtained to be observed with atomic force microscope.Results The control group vascular endothelial cells(VECs)were fusiform in shape and aligned regularly.Their size were about 11.96 ?m?3.72 ?m and their macroaxis were in parallel with the direction of hemokinesis.VECs in the atherosclerotic group were in deformity and bigger than those of the control group.They aligned irregularly and their volumes changed to be swelled.The membrane protein of VECs in the control group was composed of many round and elliptical eminences,which were almost in the same size.and with distinct boundary lines.The membrane protein of VECs in the atherosclerosis group was composed of many irregular eminences in different size.It was vague among the eminences in which there were many holes.But the VECs of atorvastatin group were better than those of atherosclerosis group.The ultramicrostructure of the membrane surface of the atorvastatin group VECs was obviously improved.Meanwhile,the mean roughness(Ra)of membrane protein of three groups was compared.The Ra of the atherosclerosis group was significantly higher than that of the control group and the atorvastatin group(P

Objective To study the correlation of expression of DNA topoisomerase Ⅱ alpha (TOP2A)with expressions of human epidermal growth factor receptor 2 (HER2)and phosphatase and tensin homolog (PTEN)and gene mutation of phosphatidylinositol 3-kinase (PI3K)in breast cancer so as to provide reference for prognosis of the cancer and evaluation of drug efficiency.Methods This study enrolled totally 96 breast cancer patients. Tumor specimens were resected.The gene expressions of TOP2A,HER2 and PTEN were analyzed using branched DNA-liquid-chip,and PI3K gene mutation was detected by xTAG-liquid-chip.Correlations between gene expressions and gene mutation were further explored by Spearman correlation analysis so as to clarify the relationship between TOP2A and HER2 signaling pathway gene.Results Co-expression of TOP2A and HER2 was strong,and TOP2A tended to be highly expressed in the presence of high expression of HER2 (P =0.01).The expression of PTEN was not significantly correlated with the expression of TOP2A,whereas the mutation of PI3K had a positive association with the high expression of TOP2A (P =0.004).Conclusion Anthracycline drug resistance factor TOP2A may be related to the critical factors of HER2 signaling pathway,suggesting that HER2 expression and PI3K mutation may be key factors in regulation of TOP2A expression,which would provide important evidence for chemotherapeutic resistance.

Objective To investigate the growth inhibition effect of evodiamine (Evo) on renal carcinoma 786-0 cells and to explore its molecular mechanism. Methods After treated with Evo, methyl thiazolyl tetrazolium ( MTT) assay was used to detect the vitality of 786-0 cells, flow cytometry was employed to examine the cell cycle distribution in 786-0 cells, and immunoblotting was utilized to determine the expression levels of target proteins related to cell cycle progression. Results Evo remarkably inhibited 786-0 cells vitality in dose-dependent manner. Cell cycle analysis indicated that 786-0 cells were arrested in G2/M phase followed by Evo treatment. Furthermore, the results of immunoblotting showed that Evo up-regulated the protein expression levels of P53, P21 and its downstream target gene CyclinB1 in 786-0 cells. Conclusion Evo treatment can induce 786-0 cell cycle G2/M arrest, and its underlying mechanism might be dependent on the P53/P21 signal pathway.

Objective To investigate the potential role of Lycium bararum polysaccharide (LBP) with or without interferon -inducible protein 10 ( CXCL10) in inducing dendritic cells ( DC) functional maturation by monitoring the alteration of cytokines for inducing DC maturation in peripheral blood and by detecting the expression of S-100 protein in tumor tissue, thus to reveal its mechanism of inhibiting experimental liver cancer. Methods H22 bearing mice model was established. The mice were randomized into model group, LBP group (50 mg/kg, ig), CXCL10 (right axillary subcutaneous injection of 15 μg/kg), LBP + CXCL10 group (LBP 50 mg/kg, ig, and right axillary subcutaneous injection of CXCL10 15 μg/kg), 5- fluorouracil (5FU) group ( intraperitoneal injection of 12mg/kg) , 12 mice in each group. The mice were administered the corresponding medicine once a day. After treatment for 2 continuous weeks, blood was sampled from infraorbital vein, and the tumor mass, spleen, thymus were extracted for the calculation of anti-tumor rate, thymus index and spleen index separately . The mRNA expression levels of interleukin 12 (IL-12) and tumor necrosis factor-α (TNF-α) in peripheral blood were detected by fluorescence quantitative PCR, the expression of S-100 protein in tumor tissues was detected by immunohistochemical assay. Results Compared with the model group, tumor growth in LBP group and LBP+CXCL10 group was obviously inhibited, and tumor-inhibitory rate was 55.90%, 50.91%, respectively. Meanwhile, the mRNA expression level of IL-12 was 2.94 folds higher in LBP group and 3.39 folds higher in LBP + CXCL10 group, and TNF-α mRNA expression level was 1.55 folds higher in LBP group and 4.74 folds higher in LBP+CXCL10 group than the model group, the differences being statistical significant ( P<0.05 or P<0.01). Results of immunohistochemical assay showed that S-100+DC number in LBP group and LBP+CXCL10 group was larger than that in the model group (P<0.05 ). Conclusion LBP and LBP+CXCL10 exert significant effect on inhibiting experimental liver cancer. The mechanism may be related with inducing the secretion of IL-12 and TNF-α, which plays a key role in inducing DC maturation, and with the increase of the number of DC in tumor microenvironment.

Objective To assess the early prognosis of 117 patients after carduopulmonary resuscitation (CPR) in ICU by using the markers of inflammation,Glasgow Coma Scale (GCS) and Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) scores.Methods A total of 117 CPR patients admitted between 2010 January to 2012 December were enrolled for study.Within 24 h after admission,inflammatory markers,blood routine items,hepatorenal function,electrolytes of blood were measured.The GCS and APACHE Ⅱ scores were recorded.Arterial blood gas analyses were performed at 0,12,and 24 h after hospitalization,and the 12-h and 24-h lactate clearance rates were calculated.Seven days after treatment,according to the outcomes,the patients were divided into survival group and death group,and the clinical data of two groups were analyzed.Results (1) Of them,73 patients died and 44 survived.Factor analysis showed that age,time elapsed from resuscitation to ICU admission,D-dimer,arterial oxygenation index (FiO2),arterial blood pH,arterial blood lactate concentration upon ICU admission,GCS score and APACHE Ⅱ score were significantly different between the two groups (P ＜ 0.05or P ＜ 0.01); (2) Two classification logistic regression analysis showed that D-Dimer,GCS score and APACHE Ⅱ score significantly correlated with the mortality risk of the patients in the wake of CPR with relative odds ratios of 1.000,2.091,and 0.531,respectively (P ＜ 0.05 or P ＜ 0.01) ; (3) Receiver operating characteristic curve analysis indicated that the area under the curve of GCS (0.821) and APACHE Ⅱ (0.869) had higher predictive value than D-dimer (0.655).The highest accuracy (84.6％) in predicting patient survival was achieved when the GCS score was 6.5.Meanwhile,the highest accuracy (82.1％) in predicting patient death was achieved when the APACHE Ⅱ score was 17.5.Conclusions Both GCS score and APACHE Ⅱ score has obvious correlation with the prognosis of the critically ill patients after CPR and could be used to predict prognosis at early stage.

Objective To discuss the influence of microRNA(miR)-155/miR-21 on toll-like receptor 4 (TLR4) in children with sepsis.Methods Fifty children with sepsis who were hospita-lized in Pediatric Intensive Care Unit,Shenzhen Children's Hospital,were enrolled in the study,and 15 healthy children at the same age were selected as healthy control group.Expression levels of TLR4 protein and human leukocyte antigen(HLA)-DR in CD14 + monocytes (MC) were detected by using flow cytometry,and sepsis patients were divided into 2 groups according to whether they exceeded the value of HLA-DR by 30％ or not.Expression level of programmed cell death factor 4 (PDCDM) and inositol phosphatases 1 containing SH2 (SHIP1) were detected at the same time.MC were separated by CD14 + immune magnetic bead,and expression level of miR-155,miR-21 and tumor necrosis factor-α (TNF-α),interleukin-10 (IL-10) mRNA in CD14 + MC were detected by using real-time fluorescent quantitative PCR.Results Sepsis group consisted of 27 male and 23 female,and their ages were (2.34 ± 0.79) years old,among whom 9 patients died.There were 36 patients in the HLA-DR increase group and 14 patients in the HLA-DR decrease group.Expressions ofTLR4(2.33±0.90),miR-155[(7.19±3.75) ×10 3] and TNF-α[(21.98±14.15) ×10-2 pg/L] in CD14 + MC were higher in the HLA-DR increase group than those in the HLA-DR decrease group [1.24±0.60,(4.83 ±1.17) × 10-3,(14.18±5.45) ×10-2 μg/L] and healthy control group[1.57±0.55,(3.99 ± 1.29) × 10-3,(1.61 ± 0.84) × 10 2 pg/L],and the differences were statistically significant(F =11.943,7.583,18.538,all P ＜0.05),while the expressions of miR-21 (12.10 ±5.66),IL-10[(29.74 ± 12.55) × 10-4 μg/L] in CD14 + MC were lower in the HLA-DR increase group than those in the HLA-DR decrease group[4.68 ± 2.07,(12.50 ± 5.73) × 10-4 μg/L] and healthy control group [2.39 ± 0.86,(2.04 ± 0.92) × 10-4 μg/L],and the differences were statistically significant(F =41.673,54.991,all P ＜ 0.05).The levels of SH1P1 and PDCD4 decreased in sepsis compared with healthy control group[0.70 ±0.36)vs.(1.59 ±0.48);(1.55 ±0.56) vs.(3.01 ±0.70)],and the differences were statistically significant (t =7.682,8.339,all P ＜ 0.05),but SHIP1 decreased more significantly in the HLA-DR increase group than that in the HLA-DR decrease group [(0.60 ± 0.34) vs.(0.97 ± 0.26)],and the difference was statistically significant (F =39.214,P ＜ 0.05).PDCD4 decreased more significantly in the HLA-DR decrease group than that in the HLA-DR increase group (0.94 ±0.19 vs.1.79 ±0.47),the difference was statistically significant(F =65.367,P ＜ 0.05).Conclusions Regulation imbalance of miR-155/miR-21 may be one of the reasons for abnormal expression of TLR4 in children with sepsis,and it plays a role in enlarged or inhibited expression of TLR4 in the sepsis process which results in different immune status in sepsis patients.

Objective To discuss the expression level of serum procalcitonin( PCT) and clinical val-ue in acute viral diarrhea patients. Methods A total of 186 patients with acute viral diarrhea treated in our hospital from September 2013 to September 2015 were retrospectively reviewed. One hundred and seven were male and 79 were female,of which 171 cases were infected by rotavirus and 15 cases infected by norovirus. The average age was ( 1. 29 ± 0. 89 ) years old. All patients′ blood and stool cultures were negative. The patients were divided into three groups according to the degree of dehydration and whether complicated with multiple organ dysfunction or not:severe group( complicated with severe dehydration,shock or multiple organ dysfunction,n=33),mild-moderate dehydration group(n=68) and no dehydration group(n=85). Thirty-five healthy children with the same age were enrolled as the control group. Serum PCT levels,high sensitivity CRP(hs CRP) and blood routine were detected. Results The serum PCT levels increased in 73 patients with acute viral diarrhea,8 cases>100 ng/ml,21 cases 5 to 100 ng/ml and 44 cases 0. 5 to 5. 0 ng/ml. PCT( ng/ml)[0. 36(0. 14,1. 67),hsCRP(mg/L)[3. 50(0. 70,14. 83)] and WBC( × 109/L)[9. 06(6. 79,12. 50)] levels increased in the diarrhea patients compared with those in the healthy group[0. 09(0. 05,0. 13);1. 00 (0.40,2.50);6.90(5.90,8.20)](all P < 0.05). The level of PCT in patients with dehydration [0.54 (0. 19,7. 83)]was higher than that without dehydration[ 0. 26(0. 11,0. 55)](P<0. 05) and increased in severe group[13. 69(3. 41,60. 30)] compared with in those the mild-moderate group[0. 33(0. 13,0. 89)] and no dehydration group[0. 26(0. 11,0. 55)](all P<0. 017),but there was no difference among groups in hsCRP and WBC levels ( P>0. 05 ) . Conclusion PCT can increase in pediatric patients with acute viral diarrhea,especially in those with severe dehydration,shock and organ dysfunction. Continued high levels of PCT indicates critical condition and has poor prognosis. PCT can be used as a good indicator to evaluate the severity of disease and the prognosis.