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Esophageal cancer is a common malignant tumor of the digestive tract. At present， the pathogenesis of esophageal cancer has not been fully clarified. Although surgery， radiotherapy， chemotherapy， targeted therapy， and immunotherapy have achieved good clinical results in the treatment of esophageal cancer， there are still many complications and severe adverse reactions. As an important part of traditional Chinese medicine （TCM）， in recent years， many basic experiments and clinical studies have proved that Chinese medicine has a good effect in treating esophageal cancer. At the same time， the multi-component and multi-target characteristics of Chinese medicine and unclear pathogenesis of esophageal cancer determine that there are some problems such as unclear mechanisms of Chinese medicine in preventing and treating esophageal cancer. It is necessary to start with modern medicine and reveal the mechanism of Chinese medicine in preventing and treating diseases from the aspects of molecular biology and network pharmacology. It is believed in TCM that the occurrence of esophageal cancer is mostly attributed to stagnation of liver Qi， phlegm stasis and Qi stagnation， fluid consumption and heat accumulation， the decline of healthy Qi， and the cementation of cancer toxicity. According to the literature review， Chinese medicinal compounds mainly include tonic formulae （such as Liu Junzitang）， drying and moistening formulas （such as Qigesan）， and heat-clearing formulas （such as Fufang Kushen injection）. Chinese medicinal monomers mainly include drugs potent in attacking poison and killing insects， clearing heat， activating blood and resolving stasis， and regulating Qi， which is consistent with the etiology and pathogenesis of esophageal cancer in TCM. It is also found that Chinese medicine can promote cell apoptosis and autophagy， block cell cycle， and reverse cell resistance by regulating phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）， neurogenic locus notch homolog protein （Notch）， Wnt/β-catenin， mitogen-activated protein kinase （MAPK）， nuclear factor-κB （NF-κB）， Janus kinase 2/signal transducer and activator of transcription 3 （JAK2/STAT3）， and other related signaling pathways， but there is no systematic summary. This study systematically summarized the relevant signaling pathways of Chinese medicine in regulating esophageal cancer， which is helpful to clarify the relevant mechanisms of Chinese medicine in the process of esophageal cancer occurrence， development， invasion， and metastasis， so as to provide new targets and new perspectives for the treatment of esophageal cancer and promote the modernization of TCM.
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OBJECTIVE To investigate the intervention effect and potential mechanism of breviscapine on hepatic fibrosis （HF） in rats based on the transforming growth factor-β（1 TGF-β1）/Smad2/extracellular signal-regulated protein kinase 1（ERK1） and Kelch-like epichlorohydrin-associated protein 1（Keap1）/nuclear factor-erythroid 2-related factor 2（Nrf2）/heme oxygenase-1（HO-1） pathways. METHODS Totally 60 rats were randomly divided into normal control group， model group， breviscapine low-dose， medium-dose and high-dose groups （5.4， 10.8， 21.6 mg/kg）， and colchicine group （positive control， 0.45 mg/kg）， with 10 rats in each group， half male and half female. Except for the normal control group， HF model of the other groups was induced by carbon tetrachloride. Subsequently， each drug group was given corresponding medicine by gavage once a day for 28 days. The liver appearance of rats in each group was observed and their liver coefficients were calculated. The levels of alanineaminotransferase （ALT） and aspartate aminotransferase （AST）in serum， those of ALT， AST， superoxide dismutase （SOD），malondialdehyde （MDA） and glutathione peroxidase （GSH- Px） in liver tissue were detected. The liver tissue inflammatory and fibrotic changes were observed. The protein and mRNA expressions of TGF-β1， Smad2， ERK1， Nrf2， Keap1 and HO-in liver tissue were detected. RESULTS Compared with the normal control group， the model group showed large areas of white nodular lesions in the liver， obvious inflammatory cell infiltration and collagen fiber deposition. The body weight， the levels of SOD and GSH-Px in liver tissue， the protein and mRNA expressions of Nrf2 and HO-1 were significantly lowered in the model group （P＜0.05）； the liver coefficient， the percentage of Masson staining positive area， ALT and AST levels of serum and liver tissue， MDA level of liver tissue， the protein and mRNA expressions of TGF-β1， Smad2， ERK1 and Keap1 were significantly increased （P＜0.05）. Compared with the model group， the liver lesions of rats in each drug group were improved， and the above quantitative indexes were generally reversed （P＜0.05）. CONCLUSIONS Breviscapine has a good intervention effect on HF rats， which may be related to inhibiting TGF-β1/Smad2/ERK1 pathway for anti-fibrosis and regulating Keap1/Nrf2/HO-1 pathway to inhibit oxidative stress.
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Objective:To investigate the cytopathic effect of amino acids 86-175 of rotavirus non-structural protein 4 (NSP4 86-175) on rat cardiomyocytes and the possible mechanism. Methods:Rat H9C2 cardiomyocytes were treated with NSP4 86-175 protein. Changes in the growth and morphology of the cells were observed. The activity of LDH in the cell culture medium was detected. Fluo-3AM was used to label intracellular free calcium ions and the concentrations of calcium ions in rat cardiomyocytes with and without NSP4 86-175 treatment were detected by confocal laser microscopy. The expression of Bax, Bcl-2, caspase-3, 78 kDa glucose-regulated protein (GRP78), C/EBP homologous protein (CHOP) and caspase-12 at mRNA level was detected by real-time PCR. The expression of caspase-3, caspase-8, caspase-9, GRP78, CHOP and caspase-12 at protein level was detected by Western blot. Results:Normal cardiomyocytes showed a typical myoblast-like morphology, presenting as spindle-shaped cells with clear boundaries. Obvious cytopathic effect, vacuolar degeneration, shriveled and rounded cells, and cell fragmentation were observed after the treatment with purified NSP4 86-175 protein. The activity of LDH in cell culture medium was enhanced by NSP4 86-175 protein. NSP4 86-175 protein also enhanced the fluorescence of the calcium ions in rat cardiomyocytes, promoted cell apoptosis, up-regulated the expression of apoptotic factors including caspase-3, caspase-8, caspase-9 and Bax-2, and increased the expression of classical markers of endoplasmic reticulum stress such as GRP78, CHOP and caspase-12. Conclusions:NSP4 86-175 had a cytopathic effect on rat cardiomyocytes, which might be related to the induced intracellular calcium overload, endoplasmic reticulum stress, apoptosis and necrosis. These results might be used as theoretical reference for further study on rotavirus infection and myocardial injury.
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Objective:To construct a mutant strain of hypervirulent Klebsiella pneumoniae NTHU-K2044 with hfq gene deletion and to analyze its biological characteristics. Methods:The hfq gene of NTUH-K2044 was knocked out by homologous recombination technology to construct △ hfq mutant strain. Its biological characteristics including growth rate, environmental stress tolerance, biofilm formation, capsular polysaccharide synthesis, resistance to neutrophil phagocytosis and lethality to Galleria mellonella larvae were analyzed by comparing with the wild-type strain using phenotypic experiments. Results:The △ hfq mutant strain of hypervirulent Klebsiella pneumoniae NTHU-K2044 was successfully constructed. Phenotypic experiments showed that the △ hfq mutant strain had significantly slower growth rate, smaller colonies and decreased hypermucoviscosity. Its growth was significantly inhibited under different environmental stress conditions such as pH9, pH5.5, 0.7 mmol/L SDS, 5% NaCl, 0.1% H 2O 2 and high temperature of 50℃. In terms of virulence and pathogenicity, the △ hfq mutant strain showed decreased ability to form biofilm and capsule, significantly down-regulated expression of magA and rmpA genes required for capsule synthesis, lower survival rate in the neutrophil bactericidal test and obviously reduced lethality to Galleria mellonella larvae. Conclusions:As a RNA chaperone, Hfq protein could participate in post-transcriptional regulation and play an important role in regulating the physiology, environmental adaptability and virulence of hypervirulent Klebsiella pneumoniae. This study provided reference for further study on hypervirulent Klebsiella pneumoniae.
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OBJECTIVE To study the effects of Tibetan medi cine Shanhu qishiwei pill in lowering blood lipid of hyperlipidemia（HLP）model rats ，and to explore its mechanism primarily. METHODS According to their body weigh ，60 SD rats were randomly divide into normal group ，model group ，simvastatin group （positive control ，20 mg/kg）and Shanhu qishiwei pill low-dose，medium-dose and high-dose groups （50，100，200 mg/kg），with 10 rats in each group. Normal group was given conventional diet ，and other groups were given high-lipid diet to induce HLP model ，for consecutive 4 weeks. Administration groups were given relevant medicine intragastrically at the same time of modeling ；normal group and model group were given equal volume of normal saline intragastrically ，once a day ，for consecutive 4 weeks. After last administration ，the serum levels of TC ，TG， LDL-C and HDL-C were determined ；pathological changes of liver tissue were observed ；the protein expressions of AMPK ， p-AMPK，LKB1，and HMGCR in liver tissue were detected in each group. RESULTS Low-dose，medium-dose and high-dose of Shanhu qishiwei pill could significantly reduce the serum levels of TC ，TG and LDL-C and protein expression of HMGCR in liver tissue（P＜0.05），while significantly increased serum level of HDL-C ，phosphorylation level of AMPK ，protein expression of LKB 1 in liver tissue in HLP model rats （P＜0.05）；the pathological changes of liver tissue in HLP model rats were improved to different extents. CONCLUSIONS Shanhu qishiwei pill can reduce the blood lipid level of HLP model rats ，and its mechanism may be related to inhibiting the transmission of LKB 1/AMPK signal pathway and regulating lipid metabolism.
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OBJECTIVE：To study the improve ment effect of salvianolate on renal interstitial fibrosis （RIF）model rats and its possible mechanism. METHODS ：Totally 50 male SD rats were randomly divided into normal group ，model group ，losartan group （positive control group ，9 mg/kg）and salvianolate low-dose and high-dose groups （18，36 mg/kg）according to body weight ，with 10 rats in each group. Except for normal group ，other groups were given adenine 250 mg/kg intragastrically to establish RIF model. After modeling ，administration groups were given relevant medicine intragastrically ，and normal group and model group were given constant volume of normal saline intragastrically ，the volume was 10 mL/kg，once a day ，for consecutive 30 days. After last medication，the serum levels of creatinine （Scr），urea nitrogen （BUN）and 24 h proteinuria （24 h UPro ）were detected by ELISA. HE staining and Masson staining were used to observe the histopathological characteristics and fibrosis of the kidney. The degree of renal tubular injury and glomerulosclerosis were scored ，and the percentage of positive staining area of renal tissue was calculated ； immunohistochemistry and Western blot assay were used to determine the protein expression of Wnt 5a，Wnt5b，and β-catenin. RESULTS：Compared with normal group ，Scr，BUN and 24 h UPro levels ，renal tubular injury score ， glomerulosclerosis score ， the percentage of positive staining area in renal tissue ，the protein expression of Wnt 5a and β-catenin were increased significantly in model group （P＜0.05），while the expression of Wnt 5b protein was decreased significantly （P＜0.05）. Pathological changes such as mesangial hyperplasia ，fibrous tissue increase and inflammatory cell infiltration were observed under microscope. Compared with model group ，above indexes of rats were improved significantly in losartan group ，salvianolate low-dose and high-dose groups （P＜ 0.05），and the effect of salvianolate had dose-dependent trend. CONCLUSIONS ：Salvianolate has the improvement effect on RIF model rats induced by adenine ，and its mechanism may be related to inhibition of Wnt/ β-catenin signal pathway.
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Objective:To analyze the data of clinical characteristics, hospitalization actual cost and hospital stay of inpatients with lymphoma, and to provide references for local epidemiological research, hospital optimization management, and appropriate resource allocation.Methods:The quadratic programming was performed based on hospital information management system and case system, as well as the information from inpatients with lymphoma in Shandong Tai'an Central Hospital between January 1, 2004 and November 30, 2019, and then they were derived and collected into the SQL server database. The gender, onset age, age distribution, and hospital stay of all inpatients with lymphoma were retrospectively analyzed. The patients were divided into the young group (< 45 years old), middle-aged group (≥45 years old and < 60 years old) and elderly group (≥ 60 years old) according to the age. The hospitalization cost and hospital stay of these inpatients in different age groups were collected and compared.Results:The number of admissions for inpatients with lymphoma between January 1, 2004 and November 30, 2019 was 3 866, accounting for 0.38% (3 866/1 011 627) of the total inpatient admissions. There were 817 newly-diagnosed lymphoma inpatients, including 499 males and 318 females (male to female ratio was 1.57:1). The median onset age of the newly-diagnosed lymphoma inpatients was 58 years old (7 months to 86 years old). Among the newly-diagnosed lymphoma patients, the proportion of them with age ≥ 45 years old was increased, and peaked at 60-65 years old, and decreased after 75 years old. There were 180, 270 and 367 cases in the young group, middle-aged group and elderly group respectively; the median hospital stay was 13.5 d, 13.0 d and 14.0 d, respectively (χ 2 = 0.419, P = 0.811). The median total cost of hospitalization was 9 846.39 yuan, 12 232.16 yuan and 14 022.36 yuan, respectively (χ 2 = 9.443, P = 0.009); and the median drug cost was 3 423.37 yuan, 3 523.62 yuan and 3 654.54 yuan, respectively (χ 2 = 2.202, P = 0.333); the ratio of median drug cost to the total cost was 44.80%, 40.49% and 34.58%, respectively (χ 2 = 6.512, P = 0.039). Conclusions:The number of lymphoma inpatients in Shandong Tai'an Central Hospital is on the rise. The incidence of inpatients with lymphoma is increased annually with older onset age. The total cost of inpatients was increased with the aging, but it has no obvious relevance with hospital stay. Middle-aged and elderly people are the target populations for disease control and prevention. Measures including screening, early diagnosis and early treatment should be enhanced.
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Objective To investigate the capsular polysaccharide (CPS) serotypes and molecular characteristics of carbapenem resistant hypermucoviscous Klebsiella pneumoniae (CR-HMKP) and study the possible mechanism of carbapenem resistance. Methods A retrospective study was conducted on 18 nonduplicate CR-HMKP strains which were collected from the First Affiliated Hospital of Nanchang University from 2012 to 2016. The clinical data were retrieved from medical records. The capsular serotypes, resistance genes and virulence factors were detected by polymerase chain reaction and DNA sequencing. Antimicrobial susceptibility testing was determined on VITEK 2 compact system. The CR-HMKP strains were characterized molecularly by using PCR, multilocus sequence typing (MLST) and pulsed field gel electrophoresis (PFGE). Modified carbapenem inactivation method was used to screen carbapenemase-producing strains. Plasmid conjugation transfer experiments were carried out to study transmission of carbapenem resistance. Results Eighteen (2.7％) CR-HMKP isolates were identified, which belonged to 4 serotypes, including wzi128-K1 (n=1), wzi206-K57 (n=1), wzi2-K2 (n=2), and wzi64-K14.64 (n=14). PCR and sequencing analysis identified blaNDM-1 gene in 2 CR-HMKP strains, blaKPC-2 gene in 17 strains, qnrS1 gene in 18 strains, blaCTX-M-3 gene in 3 strains, blaCTX-M-14 gene in 18 strains, blaTEM-1 gene in 16 strains, blaSHV-12 gene in 17 strains, and rmtB in 5 strains. All the 18 CR-HMKP strains carried virulence-associated genes, including rmpA (88.9％, 16/18), magA (5.6％, 1/18), iroN (83.3％, 15/18), aerobactin (27.8％, 5/18), rmpA2 (66.7％, 12/18) and mrkD (100％, 18/18). Three sequence types (STs) were identified by MLST, including ST11 (15 strains), ST86 (2 strains), and ST412 (1 strain). PFGE resulted in three major PFGE clusters, of which cluster A corresponds to ST1 isolates, and cluster B corresponds to ST86 isolates, and cluster C corresponds to ST412 isolates. All the blaKPC-2- positive strains belonged to ST11. Plasmid conjugation was successful in 5 (27.8％) of the 18 CR-HMKP isolates. Conclusions wzi64-K14.64 is the predominant capsule serotype of the CR-HMKP strains in this hospital. KPC-2 gene conjugationmay contribute to the emergence of CR-HMKP isolates. In addition, CRHMKP strain may be the highly prevalent ST11, and highly virulent CPS serotypes harboring K1/K2.
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Objective To investigate the antimicrobial resistant mechanisms of high level carbapenem resistant Klebsiella pneumoniae infection of burn patients .Methods A retrospective study was conducted on totally 18 non-repetitive high level CR-KP which were isolated from burn patients hospitalized between July 2014 and June 2015.MIC of antibiotics were determined by using the GN 13 cards and agar dilution method.The specific PCR and DNA sequence analysis were performed to confirm the β-lactamase type.Plasmid conjugation transfer experiments and southem hybridization were applied to study the mode of carbapenem resistance transmission .Outer membrane proteins ( Omps) were isolated and examined by PCR and ( sodium dodecyl sulfate polyacrylamide gel electrophores ) SDS-PAGE.Pulsed-field gel electrophoresis( PFGE ) and Multilocus sequence typing ( MLST ) was used to determine the genotypes . Results Susceptibility of antimicrobial agents indicated that all these strains with multiple drug resistance . The resistance rate to piperacillin/tazobactam, ceftriaxone, levofloxacin, ciprofloxacin, gentamicin, tobramycin, cefotetan, ceftazidime, cefepime, aztreonam, imipenem, and meropenem was 100％ (18/18).Moreover, the resistance rate of CR-KP isolates to amikacin was 72.2％ ( 13/18 ) , compound sulfamethoxazole was 61.1％(11/18), tigecycline was 0％(0/18).Conjugation study with Escherictda coli J53 resulted in the transfer of significant reduced carbapenem susceptibility from donors (MICs increased at least 8-fold).By PCR, eighteen strains of Klebsiella pneumoniae carried NDM-1 gene, 5 strains carried KPC-2 gene.The blaNDM-1 was transferable by plasmids.Southern blot hybridization indicated that the blaNDM-1 gene was located on plasmid in size of 46 kb.The plasmid belonged to incompatibility group IncX 3.Seven types of CR-KP were detected by PFGE.In addition, MLST assigned them to sequence type ( ST)11, ST395, ST17, ST37, ST263, ST14 and ST76 types.SDS-PAGE and ompK35/36 genes sequence analysis of Omp indicated that there was absence of outer membrane proteins OmpK 36 in ST11, ST395, ST37 strains.However, the other STs strains expressed lower quantities of OmpK 36.Conclusions High level carbapenem resistance in K.pneumoniae causing infection in burn patients is attributable to production of plasmid-mediated metallo· β-laetamase NDM-1 combined with porin OmpK36 deficiency or low expression .The K.pneumoniae with NDM-1 and KPC-2 carbapenemase were detected .
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To investigate the protective effect of the seed oil of Abelmoschus esculentus on gastric ulcer,two acute gastric ulcer mice models were established by intragastric administration of aspirin or absolute ethanol,respectively.Clinical index of ulcer area,ulcer index,gastric volume,gastric pH value,free acidity,total acidity,and histopathological assessment were measured to evaluate the injuries of gastric ulcer and the protective effect of okra seed oil In order to comprehensively uncover the possible underlying mechanism,a series of biochemical assays were also performed,including serum TNF-α,IL-6,IL-10 and Tbil,NO,MPO and SOD in the stomach included.Moreover,the ALT,AST and ALP in the liver of mice were also tested to evaluate the possible hepatic toxicity of the seed oil.The results indicated that the seed oil of A.esculentus exerted protective effect in ethanol-induced gastric ulcer mice by reducing the ulcer area and ulcer index,declining the free and total acidity,and increasing the pH value of gastric content.Histopathological observation showed the gastric mucosa of the acute gastric ulcer mice induced by alcohol was incomplete and severely damaged,with submucosal edema and nuclear pyknosis,as well as glandular structure disappearing,compared with that of normal mice.What's more,a number of inflammatory cell infiltration occured in the gastric mucosa of alcohol-model mice,with messes of neutrophils,lymphocytes,eosinophils and plasma cells.Okra seed oil could improve the damaged structure of the gastric mucosa and gland caused by ethanol,but could not ameliorate the condensation of nucleus and infiltration of inflammatory cells.Biochemical analysis revealed that the seed oil of A.esculentus could counteract the damage induced by ethanol via decreasing Tbil and TNF-α in serum,decreasing NO and myeloperoxidase,and increasing SOD in stomach.Meanwhile,okra seed oil exhibited protective effect in aspirin-induced gastric ulcer mice by increasing the gastric content pH,and reducing free and total acidity.Compared with the control group,the gastric mucosa of aspirin-model group showed multifocal coagulation necrosis,sheet edema and infiltration of inflammatory cells by histopathological assessment.Compared with the aspirin-model group,the soybean oil group and okra seed oil group could ameliorate the inflammatory cell infiltration.Biochemical analysis revealed that okra seed oil could counteract the injury induced by aspirin via decreasing TNF-α and IL-6,and increasing IL-1O in serum,decreasing NO and MPO and increasing SOD in stomach.In a word,the okra seed oil exerted protective effect on acute gastric ulcer by anti-inflammation,anti-oxidation and hepatocyte protection.The okra seed oil deserves further development and utilization.
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Objective To evaluate the performance of VITEK 2-Compact GN13 methods for testing imipenem susceptibility of Klebsiella pneumoniae and assess the reliability of its Advanced Expert System (AES).Methods A retrospective study was conducted with a total of 157K. pneumoniae strains, which were isolated from blood and intra-abdominal infections in the First Affiliated Hospital of Nanchang University from 2014 to 2015. Thein vitro minimum inhibitory concentration (MIC) values of imipenem were determined by disc diffusion, VITEK 2-Compact GN13 and broth microdilution methods, respectively. Categorical agreement (CA) rates of disc diffusion and VITEK 2-Compact GN13 methods were determined using broth microdilution as reference method. The genes encoding ESBLs and carbapenemase were screened by PCR and sequencing analysis. The phenotypic confirmatory tests such as modified Hodge test, PCR and DNA sequencing were used to confirm the resistance mechanism and evaluate the reliability of AES in interpreting the imipenem susceptibility of K. pneumoniae.Results Among the 157 isolates, 64 and 8 were identified as resistant and intermediate strains by broth microdilution method, respectively; 52 and 10 were tested as resistant and intermediate strains by disc diffusion method, respectively; 54 and 13 were determined as resistant and intermediate strains by VITEK 2-Compact GN13 method, respectively, while 70 and 3 were judged as resistant and intermediate strains by VITEK 2-Compact GN13 method plus AES validation. The CA of disc diffusion and VITEK 2-Compact GN13 methods compared with broth microdilution method were all higher than 90 %. However, the major error (ME) rate was 3.8 % and very major error (VME) rates were all 0.6 % in imipenem susceptibility testing by VITEK 2-Compact GN13 and disc diffusion. The imipenem susceptibility of 16 strains were modified by the AES, which eliminated 0.6 % VME, but increased major error by 1.3 % and minor error by 1.9 %. Phenotypic confirmatory tests showed that 75 % (12/16) of these strains were validated as producers of both ESBLs and carbapenemase, which was consistent with the result of AES validation. PCR and DNA sequencing analysis proved that 62.5 % (10/16) of these strains produce IMP-4/KPC-2 /NDM-1 and ESBLs.Conclusions Both disc diffusion and VITEK 2-Compact GN13 methods can be used for testing the imipenem susceptibility of K. pneumoniae isolates with reliable and accurate results. Attention should be paid to the possibility of ME and VME when testing imipenem susceptibility. The VME can be avoided by the AES mechanism. However, AES intervention will increase ME and minor error, which may be associated with decreased expression of carbapenemase.
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Objective To explore the expression and significance of 25-hydroxy vitamin D [25(OH)D] in rheumatic disease patients,and its association with the risk of cardiovascular disease (CVD).Methods The level of 25(OH)D,bone mineral density (BMD) test and clinical indicators of 134 rheumatic disease cases were analyzed retrospectively.T test was used for statistical analysis.Results Patients who were combined with CVD and rheumatic disease,the level of 25-hydroxy vitamin D was (13±8) ng/ml,who were without CVD the level was (14±8) ng/ml.There was no significant difference between two groups(t=0.638,P＞0.05).But patients who were complicated with CVD and rheumatic diseases,their TC,TG,high-density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C),apolipoprotein (Apo) A,ApoB and lipoprotein levels were higher than patients who were without CVDs (t=3.314,3.397,3.092,2.159,3.621,2.011,2.066,both P＜0.05),and their BMD was lower than patients who were without CVDs (t=-16.131,P＜0.05).Conclusion To a certain extent,serum 25(OH)D levels of patients with rheumatic diseases is associated with CVD.
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@#A qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(HPLC-QTOF-MS)was developed for identification of major constituents in process residue of tripterygium glycosides. The HPLC-QTOF-MS assay was performed on a Zorbax SB-C18 column(4. 6 mm × 50 mm, 1. 8 μm)with the mobile phase consisting of acetonitrile and water containing 0. 2% formic acid in gradient mode. Positive ion mode was used for TOF-MS. According to the accurate molecular weight, MS fragment pathway, comparison with the retention time of reference compounds, total 30 compounds, including fifteen alkaloids, ten diterpenoids, four triterpenoids and an unsaturated fatty acid were identified or tentatively characterized in process residue of tripterygium glycosides. This study may be helpful to the comprehensive exploitation and utilization of process residue of tripterygium glycosides.
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Objective To investigate the molecule phenotype, epidemiology, and resistance genes of the New Delhi metallo- β-lactamase-1 ( NDM-1 ) producing Klebsiella pneumoniae ( K. pneumoniae ) . Methods Retrospective study was made on one hundred and ten non-repetitive carbepenem-resistant K. pneumoniae clinical isolated strains, which were collected from January 2011 to December 2012 in our hospital. The minimal inhibitory concentrations ( MICs ) of antibiotics were tested by the GN13 cards of BioMerieux Company. Modified Hodge test were used for the detection of carbapenemases. The blaNDM-1 encoding gene and linkage of ISAba125-NDM were detected by PCR method. The purified PCR products were cloned and sequenced. The homology of the K. pneumoniae were analyzed by the multilocus sequence typing ( MLST ) . Plasmid conjugation experiment and curing method were used to study the transfer of bacterial resistance. The Fisher′s exact probability test was used to compare the data. Results 13% NDM-1-producing K. pneumoniae were detected and confirmed as blaNDM-1 by sequencing (14/110). The resistance rates of the 14 NDM-1-producing K. pneumoniae strains to imipenem, meropenem, ertapenem, ciprofloxacin, levofloxacin, amikacin, and aztreonam were 14/14, 14/14, 13/14, 10/14, 9/14, 5/14, and 11/14. Meanwhile, the positive rate of ISAba125-NDM linkage of those 14 NDM-1-producing K. pneumoniae strains was 14/14. The E. coli J53 transconjugants, whose MICs of imipenem, meropenem, and ertapenem were increased by 4 to 64 times, were blaNDM-1 gene and ISAba125-NDM linkage positive. In addition, it was showed that the blaNDM-1 gene and ISAba125-NDM linkage were located on a plasmid with a size of approximately 65 000 bp. Conclusions The NDM-1 producing K. pneumoniae strains in this study were resistant to many commonly used antibiotics, however, the resistance rate to aminoglycoside and aztreonam were relatively low. The carbapenemase-resistant genotype spread by blaNDM-1 carried plasmid. Attention should be paid to its easily transmissible feature among the strains in clinic. The insertion sequence ISAba125 may be involved in the blaNDM-1 gene mediated carbapenemase-resistant genotype.
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AIM:To explore the change of antithrombin Ⅲ( AT-Ⅲ) in the patients with atherosclerotic cere-bral infarction .METHODS:Chromogenic substrate assay was used to measure the activity of AT-Ⅲ in 55 patients with atherosclerotic cerebral infarction and 55 healthy controls , and the correlation analysis was applied to determine the AT-Ⅲactivity with the severity of damage in central nervous system and general biochemical parameters .The levels of TNF-αand IL-6 in the plasma were detected by ELISA .Immunocomplex in the plasma was measured by enzyme immunoassay (EIA). The number and phenotype of the monocytes in peripheral blood were analyzed by flow cytometry .ELISA was also applied to determine the secretion of TNF-αand IL-6 from the monocytes after the stimulation of immunocomplex .The expression of AT-Ⅲin human brain vascular endothelial cells after the stimulation of TNF-αand IL-6 was observed by Western blotting . RESULTS:The activity of AT-Ⅲsignificantly decreased in the patients with atherosclerotic cerebral infarction , and nega-tively correlated with the damage degree of nervous system function , systolic pressure , diastolic pressure , glucose , choles-terol, triglyceride, low-density lipoprotein cholesterol and homocysteine , while positively correlated with high-density lipo-protein.In addition, the plasma levels of TNF-αand IL-6 increased significantly , accompanied with the enhancement of immunocomplex level .The numbers of CD14 + CD16 + and CD14 + CD32 + monocytes in peripheral blood were not changed , while CD14 +CD64 +monocytes increased obviously .The secretion of TNF-αand IL-6 by monocytes were signifi-cantly enhanced after stimulated with immunocomplex , while the protein expression of AT-Ⅲ in the human brain vascular endothelial cells was down-regulated after co-incubated with TNF-αor IL-6.CONCLUSION:Decreased AT-Ⅲactivity in the patients with atherosclerotic cerebral infarction is one of the risk factors of cerebral infarction , and related with the dis-ease severity .The production of pro-inflammatory cytokines through immunocomplex from CD 14 +CD64 +monocytes may be involved in the mechanism .Improvement of AT-Ⅲactivity may protect against cerebral ischemia .
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Objective To observe the effect of hemoperfusion on micro-inflammation in patients with maintenance hemodialysis (MHD).Methods Fifty MHD patients (MHD group) and 25 healthy volunteers (control group) were involved in this study.The MHD patients were divided into two groups by random digits table:hemoperfusion combined with hemodialysis group (HP + HD group,25 cases) and hemodialyais group (HD group,25 cases).The plasma levels of micro-inflammatory cytokines including high-sensitive C-reactive protein (hs-CRP),interleukin-6 (IL-6) and tumor necrosis faetor-α (TNF-α ) were measured before and after treatment.Results The plasma levels of hs-CRP,IL-6 and TNF-α in MHD group were significantly higher than those in control group [(6.72 ± 2.63) mg/L vs.(1.35 ± 0.92) mg/L,(348.83 ± 64.41) ng/L vs.(54.49 ±22.47) ng/L,(7.52 ± 3.17) ng/L vs.(2.53 ±0.88) ng/L](P＜0.05).There was no significant difference in the plasma levels of hs-CRP,IL-6,TNF-α before treatment between HD group and HP+HD group (P＞0.05),Compared with those in HD group,the plasma levels of hs-CRP,IL-6 and TNF-α after treatment in HP+HD group were significantly decreased [(4.78 ±2.49) mg/L vs.(6.89 ±2.69) mg/L,(260.54 ±56.72) ng/L vs.(357.14 ±56.37) ng/L,(5.36 ±2.41) ng/L vs.(7.49 ±2.87) ng/L] (P <0.05).Conclusion Hemoperfusion improves micro-inflammation in patients with MHD.
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The results of RT-PCR and immunohistochemistry showed that the expressions of vascular endothelial growth factor (VEGF) and its receptor ( flk-1 ) in the renal cortex of insulin-resistant rats during the phase of normal blood glucose were significantly increased, which were decreased by telmisartan. The result suggests that telmisartan may ease kidney damage via decreasing VEGF and flk-1 expressions.