OBJECTIVE To investigate the effect mechanism of andrographolide （Andro） on neuropathic pain （NP） in rats. METHODS Rats were randomly separated into sham operation group， model group， Andro low-dose （1 mg/kg）， Andro medium-dose （5 mg/kg） and Andro high-dose （10 mg/kg） groups， and sodium ferulic acid （150 mg/kg） group， with 12 rats in each group. Except for sham operation group， other groups used the chronic sciatic nerve compression injury method to induce NP model. After modeling， each group was given relevant dose of Andro intrathecally or sodium ferulate intragastrically. The sham operation group and model group were given a constant volume of normal saline once a day for 14 consecutive days. The mechanical withdrawal threshold （MWT） and thermal withdrawal latency （TWL） of rats were detected in each group after 7 and 14 days of administration. After the last medication， the serum levels of interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， prostaglandin E2 （PGE2）， and substance P （SP） in rats were detected in each group， and the pathological morphology of spinal cord tissue was observed. mRNA and protein expressions of ionized calcium binding adaptor molecule-1 （Iba-1）， growth arrest specific protein 6 （Gas6）， and Axl in spinal cord tissue were determined. RESULTS Compared with model group， MWT and TWL after 7 and 14 days of the administration， the mRNA and protein expressions of Gas6 and Axl after the last medication were all increased significantly in administration groups （P＜0.05）， while the levels of IL-6， TNF-α， PGE2 and SP， mRNA and protein expressions of Iba-1 were all decreased significantly （P＜0.05）； pathological injuries such as the disordered arrangement of spinal cord neurons and dilation and congestion of capillaries had been alleviated to varying degrees. Compared with sodium ferulic acid group， there was no statistically significant difference in the above indicators in the Andro high-dose group （P＞0.05）. CONCLUSIONS Andro may inhibit inflammatory response by activating the Gas6/Axl signaling axis， thereby alleviating NP.

Abstract: Objective To express Mycobacterium tuberculosis Rv2626c protein in Escherichia coli (E. coli) and study the antigenicity of the purified recombinant Rv2626c protein. Methods The amino acid sequence of Rv2626c protein from Mycobacterium tuberculosis H37Rv strain (accession number: CCP45424.1) in GenBank was retrieved and converted into the corresponding DNA sequence according to the codon preference of E. coli. This DNA sequence was synthesized and cloned into pET24a(+) plasmid to construct pET24a(+)-Rv2626c recombinant plasmid. This plasmid was transformed into E. coli BL21(DE3) cells, and the expression of Rv2626c protein was induced under various conditions of isopropyl β-D-thiogalactopyranoside (IPTG) concentrations, temperature, and period. The recombinant Rv2626c protein was identified by SDS-PAGE and Western Blot. The recombinant Rv2626c protein was purified by nickel chelate affinity chromatography and used to immunize violet blue rabbits to prepare anti-Rv2626c anti-serum. The specificity and titer of the serum were respectively detected by Western Blot and enzyme-linked immunosorbent assay (ELISA). Results The recombinant plasmid pET24a(+)-Rv2626c was successfully constructed. SDS-PAGE analysis showed that recombinant Rv2626c was expressed in the recombinant plasmid transformed E. coli with IPTG induction, with a molecular weight of about 14 500, and the size was consistent with the expectation. The optimal expression condition for recombinant Rv2626c protein was at 31 ℃ with 1.0 mmol/L IPTG for 6 hours. The target protein was mainly present in a soluble form, which was consistent with the results of Western blot. The hyperimmunized serum with recombinant Rv2626c protein vaccination showed good specificity, with a titer of 1∶ 256 000 detected by ELISA. Conclusions Mycobacterium tuberculosis Rv2626c protein is successfully expressed in E. coli, and the purified protein has good purity and antigenic activity, laying the foundation for further reveals of its biological functions.

【Objective】 To explore the risk factors of transfusion-related acute lung injury (TRALI). 【Methods】 The clinical symptoms, signs, imaging examinations, and laboratory test results of two patients with TRALI after blood transfusion were retrospectively analyzed, and human leukocyte antigen (HLA) genotyping of the patient and HLA antibodies typing of the plasma donors were performed. 【Results】 The clinical manifestations and laboratory parameters of two patients were consistent with those of TRALI after blood transfusion. After timely clinical respiratory support treatment, all patients were improved. Blood donors produced high titers of HLA-Ⅱ antibodies after pregnancy, including antibodies that specifically recognize the patient′s HLA antigen. 【Conclusion】 Two patients developed TRALI after platelet transfusion from a female blood donor, which was caused by HLA-Ⅱ antibodies.

Objective: To analyze the association between different sleep behaviors and overweight and obesity of junior high school students in Yangzhou City, and to provide a basis for policies and interventions related to adolescent health management. Methods: A total of 1 589 students in grades 7-9 from two middle schools in Yangzhou City were selected using the cluster sampling method and were administered with sleep time, bedtime, social jetlag difference, and sleep habits. Results: Totally 64.38% were sleep deprived during the school days, 86.78% went to bed too late, 46.51% had a social jetlag of ≥1 h, and 37.44% took a nap every day(Incluldes holidays and school days). Social jetlag length was statistically different between grades( F =6.97, P < 0.01 ). Girls[(0.95±0.65)h] shown significantly higher social jetlag than the boys[(0.76±0.59)h]( t=6.19, P <0.01). Later bedtime on weekends, later wake up time on weekends and poor sleep behavior were risk factors for overweight and obesity in junior high school students( OR=1.20, 1.14, 1.04, P <0.05). Conclusion Junior high school students had less sleep and later bedtimes with the increase of grade, and weekend bedtimes,wake up times and poor sleep behavior were independently associated with the risk of overweight and obesity in junior high school students. Parents and schools should be instructed to pay attention to their sleep health and carry out adolescent sleep health guidance.

Background and purpose:As a tumor suppressor gene, ribosomal S6 kinase 4 (RSK4) plays important roles in inhibiting cell proliferation, migration and inducing cell apoptosis. However, the proteins interacting with RSK4 are still unknown. This study aimed to screen proteins interacting with RSK4 in breast cancer cell line MDA-MB-231 by lfag-tag affnity puriifcation and LC-MS/MS (liquid chromatography/mass spectrometry).Methods:The pcDNA3.1/EGFP-RSK4-Flag eukaryotic expression vector was constructed by inserting full lengthRSK4 gene into vector pcDNA3.1/EGFP-Flag. And then the recombinant plasmids were transferred into MDA-MB-231 cells. Real-timelfuorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot were used to detect the expression of RSK4 in MDA-MB-231 cells. Affnity puriifcation and LC-MS/MS were applied to screen proteins interacting with RSK4, and the related action mechanism of RSK4 with its interacted proteins was detected based on bioinformatics gene ontology (GO) and ingenuity pathway analysis (IPA).Results:Twenty-four proteins, such as serine/threonine-pro-tein kinase 38 (STK38)/serine/threonine-protein kinase 38-like (STK38L), MOB kinase activator 2 (MOB2) and protein arginineN-methyltransferase 5 (PRMT5), were successfully identiifed by Flag-tag affnity puriifcation followed by LC-MS/MS analysis, which probably interacted with RSK4. Bioinformatics analysis of the identiifed proteins suggested the proteins interacting with RSK4 were involved in diverse biological pathways, such as apoptosis and cell migration. Conclusion:According to bioinformatics results of proteins interacting with RSK4 identiifed by affnity puriifcation and LC-MS/MS, biological networks of RSK4 are involved in apoptosis and migration in breast cancer cells.

OBJECTIVETo analyze the clinical feature and constituent ratio of adult hip fractures in Southwest China.

METHODSThe data of adult inpatients and outpatients with hip fractures treated between January 2010 and December 2011 in 11 hospitals of the Southwest China were collected and analyzed. The data includes gender, age, age distribution and fracture pattern according to AO classification.

RESULTSThere were a total of 2,833 adult hip fractures, including 1,340 (47.30%) males and 1,493 (52.70%) females, with a male-to-female incidence ratio of 1: 1.11 and a mean age of (66±18) years. The highest frequency of hip fractures was seen in the 71 to 85 years age group (42.18%, 1,195/2,833). There were 844 fractures (29.79%) in the young and middle-aged group (16-<60 years) and 1 898 fractures (70.21%) in the geriatric group (≥60 years). Men had a higher rate than women (men: 577 fractures, 68.4%) in the young and middle-aged group, while women had a higher rate than men (women: 1,226 fractures, 61.64%) in the geriatric group, with a significant difference in the sex distribution between the two groups (χ2=214.001, P<0.01). The proportion of intertrochanteric fracture (type 31-A), femoral neck (type 31-B) and femoral head fracture (type 31-C) was 46.59%, 49.74% and 3.67% respectively. The highest frequency of the sub-type in each fracture type was type 31-A2, type 31-B2 and type 31-C2.

CONCLUSIONSWomen have a higher rate than men in Southwest China. Geriatric patients are more than the young and middle-aged patients. The femoral neck fractures, intertrochanteric fractures and femoral head fractures are in descending orders according to the proportion of the three different hip fractures.

Aged ; Aged, 80 and over ; China ; epidemiology ; Female ; Femoral Fractures ; Femoral Neck Fractures ; Femur ; Femur Head ; Femur Neck ; Hip Fractures ; epidemiology ; Humans ; Incidence ; Male ; Middle Aged

The chemical constituents of Rosa chinensis Jacq were diverse ,mainly including flavonoids ,flavonoid glyco-sides ,phenolic acids ,aromatic oils ,tannins and pigments .Its extract and some chemical constituents had shown multiple phar-macological activities ,such as antitumor ,antifungal ,anti-viral ,anti-oxidation etc ..The advances in the study on chemical com-ponents and pharmacological actions of Rosa chinensis Jacq were reviewed and its application prospect was prospected .

An ultrasensitive immunoassay was developed based on As3+ and Hg2+ labeled SiO2 @ Au nanoparticles signal tags and hydride generation-atomic fluorescence spectrometry (HG-AFS) for the detection of carcinoembryonic antigen(CEA) and carbohydrate antigen 19-9 (CA 19-9) respectively. Firstly, amino SiO2@ Au NPs were synthesized for selective absorption of As3+ and Hg2+ ions respectively. Subsequently,the secondary antibody (Ab2) of CEA and CA 19-9 was respectively labeled on As3+ or Hg2+-SiO2 @ Au NPs to prepare the corresponding signal tags for CEA and CA 19-9. Based on the sandwich immunoassay scheme, the tags, two antigen and corresponding first antibodies were bio-conjugated on the bottom of 96-well plate at room temperature to form the immunocomplex. After it was dissolved in alkali solution, As3+ and Hg2+ ions were released in solution and detected by HG-AFS, which concentration was proportional with logarithms of CEA and CA 19-9. The reaction conditions were optimized and the tags were characterized. This assay was based on determination of the concentration of As3+ and Hg2+ for quantization of the corresponding CEA and CA 19-9 antigen. The assay showed a wide linear range from 0. 001 to 100. 0 μg / L for CEA and 0. 01-80 U/ mL for CA 19-9, and a lower detection limit of 0. 5 ng / L and 0. 005 U/ mL respectively. This proposed method was used in real serums samples, the results were consistence with that by ELISA. The immunoassay showed three orders of magnitude of sensitivity lower than that of ELISA, which provides a promising simultaneous immunoassay for the early diagnosis of cancer .

The target of rapamycin ( TOR) , a Ser/Thr protein kinase of PIKKs ( phosphatidylinositol kinase-related kinases ) , is the central factor of a highly conserved signaling pathway in eukaryotes , and regulates cell growth in response to nutrients , hor-mones, and stresses.It controls temporal growth by activating anabolic processes such as translation , ribosome biogenesis , protein syn-thesis, transportation of amino acid and metabolic enzymes .The advances in TOR pathway in the most pervasive human fungal patho-gen Candida albicans.

Objective To investigate the protective effects and mechanism of TG 6 on myocardial ischemia/reperfusion injury . Methods the protective effects of TG 6 on myocardial ischemia/reperfusion was investigated by setting up models of ischemia and reperfusion of rats induced by ligating the left coronary anterior descending artery in vivo,isolated rat hearts through an improved Lange-ndorff device, and hypoxia /reoxygenation injury of neonatal rat cardiomyocytes , and the serum CK,LDH,T-SOD, MDA were taken as research markers .Results TG6 significantly reduced the myocardial infarct size , decreased the activity of CK and the content of MDA in serum, reduced the activity of LDH, and increased the activity of T-SOD in vivo;TG6 obviously increased the coronary blood flow after low rate perfusion and reperfusion , decreased the content of MDA and the leakage of CK , LDH in myocardial tissue , elevated the activity of T-SOD in vitro of isolated rat hearts;TG6 had no effects on cells in normal growth condition , raised the viability of cardio-myocytes significantly, and reduced the rate of CK leakage and the content of [Ca2+]i obviously in Na2S2O4 treated cells in vitro of neonatal rat cardiomyocytes .Conclusion TG6 could effectively protect myocardial ischemia/reperfusion injury .