BACKGROUND: The emergence of fosfomycin resistance and extended-spectrum β-lactamase (ESBL) genes is a serious threat to public health and a new challenge in shigellosis treatment. The purpose of this study was to identify fosfomycin resistance and characterize β-lactamase genes in fos-carrying isolates of Shigella flexneri from patients in China. METHODS: A total of 263 S. flexneri isolates were collected from 34 hospitals in the Anhui Province of China during September 2012-September 2015 and screened for fosA3, fosA, and fosC2 by PCR amplification and sequencing. The fos-carrying isolates were then screened for β-lactamase genes. The clonal relationships between fosA3-carrying isolates, the transmissibility of fosfomycin resistance, replicon types of plasmids carrying fosfomycin resistance genes and other associated resistance genes were investigated. RESULTS: Twenty-five of the 263 isolates (9.5%) showed resistance to fosfomycin, and 18 (6.8%) were positive for fosA3. None of the isolates was positive for fosA or fosC2. Seventeen of the isolates carrying fosA3 (94%) were CTX-M producers (seven CTX-M-55, five CTX-M-14, and five CTX-M-123), while three (16.7%) were TEM producers (TEM-1).Sixteen (88.9%) fosA3-carrying isolates exhibited multi-drug resistance. The replicon types of the 13 fosA3-carrying plasmids were IncF (n=13), IncHI2 (n=3), IncIl-Ir (n=2), and IncN (n=1). CONCLUSIONS: Our results indicated that fosA3 could spread through plasmids in S. flexneri isolates, along with the bla(CTX-M) and bla(TEM), which facilitate its quick dispersal. To the best of our knowledge, this is the first report of CTX-M-123-type ESBLs in S. flexneri isolates from patients in China.
China* ; Drug Resistance, Multiple ; Dysentery, Bacillary ; Fosfomycin* ; Humans ; Plasmids ; Polymerase Chain Reaction ; Public Health ; Replicon ; Shigella flexneri* ; Shigella*

OBJECTIVETo detect the expression of transient receptor potential canonical 1 (TRPC1) in a mouse model of ozone-induced lung inflammation and explore its role in lung inflammation.

METHODSIn a mouse model of lung inflammation established by ozone exposure, the expression of TRPC1 in the inflammatory lung tissues was detected by RT-PCR, Wstern blotting and immunohistochemistry.

RESULTSCompared to the control mice, the mice exposed to ozone showed significantly increased expression level of TRPC1 mRNA and protein in the inflammatory lung tissues (P<0.05). Immunohistochemistry showed increased TRPC1 protein expressions in the alveolar epithelial cells, bronchial epithelial cells, and inflammatory cells in the inflammatory lung tissues (P<0.05). The mRNA and protein expression levels of TRPC1 were positively correlated with the counts of white blood cells, macrophages, neutrophils and lymphocytes in the bronchoalveolar lavage fluid of the exposed mice (P<0.01).

CONCLUSIONTRPC1 may play a role in ozone-induced lung inflammation in mice.

Animals ; Bronchoalveolar Lavage Fluid ; Disease Models, Animal ; Gene Expression ; Inflammation ; pathology ; Lung ; metabolism ; pathology ; Mice ; Ozone ; adverse effects ; Pneumonia ; metabolism ; pathology ; RNA, Messenger ; TRPC Cation Channels ; metabolism

OBJECTIVETo study the mechanism underlying the effect of the SOCS3 rs4969170 A/G alleles on the occurrence of insulin resistance (IR) in patients with chronic hepatitis C.

METHODSThe promoter region of the SOCS3 gene was amplified by PCR,and luciferase expression vectors were constructed and transfected into HepG2,Huh7 cell lines.The relative luciferase activity of each expression vector was assessed by the dual luciferase reporter gene assay system.Western blotting was used to detect SOCS3 protein expression in PBMCs from groups of patients with the rs4969170 AA and AG genotypes.The state of IR in eight patients was evaluated by determining their HOMA-IR.

RESULTSThe pGL3-A, PGL3-G and pGL3-control vectors showed significantly different luciferase expression in the HepG2 cells (0.121 00 ± 0.022 07,0.027 00+/-0.012 49 and 0.043 33 ± 0.005 51; F =48.068, P=0.001) and in the Huh7 cell lines (0.164 70 ± 0.007 10,0.027 33 ± 0.017 04 and 0.033 67 ± 0.014 98; F =115.137, P=0.001). The expression of SOCS3 protein was significantly higher in the rs4969170 AA genotype group than in the AG genotype group (1.22 ± 0.40 vs. 0.30 ± 0.19; t =4.149, P=0.006).The IR index of patients with the rs4969170 AA genotype and the AG genotype was 4.11 ± 2.62 and 1.47 ± 1.01 respectively.There were three patients with IR in the rs4969170 AA genotype group and one in the rs4969170 AG group. There was no statistically significant difference between the two genotype groups (t=1.881, P=0.109).

CONCLUSIONSThe SOCS3 rs4969170 A haplotype may enhance transcriptional activity of the gene promoter to regulate gene expression, thereby increasing intracellular SOCS3 protein level and ultimately interfering with insulin signaling and causing IR in patients with chronic hepatitis C.

Cell Line, Tumor ; Genes, Reporter ; Genotype ; Haplotypes ; Hepatitis C, Chronic ; Humans ; Insulin Resistance ; Luciferases ; Polymorphism, Single Nucleotide ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins

OBJECTIVETo examine the expression of cytokine-induced apoptosis inhibitor1 (CIAPIN1) in gastric cancer and normal mucosa tissues, and to investigate its effects on migration and invasion of gastric cancer cells.

METHODSImmunohistochemistry was used to detect CIAPIN1 expression in 15 samples of normal gastric mucosa tissues, 148 samples of gastric cancer tissues (42 of non-metastasis gastric cancer tissues without other organs or lymph node metastasis, 106 of metastasis gastric cancer tissues with lymph node metastasis), and 37 samples of gastric cancer lymph node metastasis tissues. Association of CIAPIN1 expression with clinicopathological characteristics of gastric cancer was analyzed by Chi-square test. SGC-7901 cell lines of high CIAPIN1 expression and low CIAPIN1 expression were established. Transwell assay was applied to detect the invasion and migration of CIAPIN1-regulated gastric cancer cells.

RESULTSPositive rate of CIAPIN1 expression in gastric cancer tissues was lower than that in normal gastric mucosa tissues (41.2% vs. 86.7%, P=0.0008), and in metastasis gastric cancer tissues was also lower than that in non-metastasis gastric cancer tissues (34.9% vs. 71.4%, P=0.0000). Furthermore, the positive rate of CIAPIN1 expression was closely related to the TNM staging of gastric cancer (TNM stage I(, II( and III( were 55.0%, 53.5% and 24.4%, respectively, P=0.0037). But there was no significant difference of positive expressive rate between metastatic gastric cancer tissues and metastatic lymph node tissues(34.9% vs. 21.6%, P=0.3700). Transwell assay showed that up-regulated CIAPIN1 expression would significantly reduce, while down-regulated CIAPIN1 expression would significantly promote the invasion and migration of SGC-7901 cells (all P<0.05).

CONCLUSIONPositive rate of CIAPIN1 expression is low in gastric cancer tissues and shows inhibition of invasion and migration of gastric cancer cells.

Cell Line, Tumor ; Down-Regulation ; Humans ; Immunohistochemistry ; Intracellular Signaling Peptides and Proteins ; metabolism ; Lymphatic Metastasis ; Neoplasm Staging ; Stomach Neoplasms ; metabolism ; Up-Regulation

No abstract available.
Aged ; Anti-Bacterial Agents/*pharmacology ; Bacterial Proteins/genetics ; Drug Resistance, Bacterial/*genetics ; Fluoroquinolones/*pharmacology ; Humans ; Male ; Microbial Sensitivity Tests ; Mutation ; Serratia Infections/*diagnosis/microbiology ; Serratia marcescens/*drug effects/genetics/isolation & purification

BACKGROUNDMany studies have shown that continuous renal replacement therapy (CRRT) could clean lactate and treat the hyper-lactatemia. On the contrary, some other studies found that filter lactate clearance only accounted for a very small part of total lactate clearance and the hemofilter's contribution to the overall lactate clearance was negligible. The objective of this study was to evaluate the effects of various doses of continuous veno-venous hemofiltration (CVVH) on plasma lactate elimination in critically ill patients.

METHODSPatients were divided into three groups according to their incipient plasma lactate concentration. Group A: lactate ≤ 2 mmol/L, group B: lactate 2-5 mmol/L, group C: lactate ≥ 5 mmol/L. Three different doses (20 ml × kg(-1)× h(-1), 35 ml × kg(-1)× h(-1) and 45 ml × kg(-1)× h(-1)) of CVVH were applied to critically ill patients who experiencing CVVH. The concentrations of plasma lactate in pre-(A), post-dialyzer (V) sites and ultrafiltrate were measured after each dosage of CVVH was carried out for 30 minutes. Rate of lactate clearance by the filter (RLC) and filter lactate clearance (FLC) and Lactate-Sieving Coefficient (LSC) were calculated under different circumstances, including different doses of CVVH and different incipient lactate levels.

RESULTSFifteen patients were enrolled and 104 blood samples were drawn and lactate concentrations were measured in this study. RLC was found increased ((9.36 ± 9.73) mmol/h, (13.92 ± 12.56) mmol/h and (16.52 ± 12.71) mmol/h, P < 0.05 respectively) with the dose of CVVH increased. RLC was also increased ((3.46 ± 1.46), (10.38 ± 5.50) and (24.53 ± 14.69) mmol/h, P < 0.05 respectively) with the incipient lactate increased. FLC was increased ((1.95 ± 0.63), (2.95 ± 0.74) and (3.45 ± 0.54) L/h, P < 0.05 respectively) with the dose of CVVH increased. There was no significant difference of LSC in different doses of CVVH and different incipient lactate levels.

CONCLUSIONSPlasma lactate can be eliminated by CVVH and different doses of CVVH affect the rate of lactate clearance in critically ill patients.

Adult ; Aged ; Aged, 80 and over ; Critical Illness ; Female ; Hemofiltration ; Humans ; Lactic Acid ; blood ; Male ; Middle Aged

BACKGROUND: Shigella is a frequent cause of bacterial dysentery in the developing world. Treatment with antibiotics is recommended for shigellosis, but the options are limited due to globally emerging resistance. This study was conducted to determine the frequency and pattern of antimicrobial susceptibility of Shigella in China. METHODS: We studied the antimicrobial resistance profiles of 308 Shigella spp. strains (260 S. flexneri, 40 S. sonnei, 5 S. boydii, and 3 S. dysenteriae) isolated from fecal samples of patients (age, from 3 months to 92 yr) presenting with diarrhea in different districts of Anhui, China. The antimicrobial resistance of strains was determined by the agar dilution method according to the CSLI guidelines. RESULTS: The most common serogroup in the Shigella isolates was S. flexneri (n=260, 84.4%), followed by S. sonnei (n=40, 13.0%). The highest resistance rate was found for nalidixic acid (96.4%), followed by ampicillin (93.2%), tetracycline (90.9%), and trimethoprim/sulfamethoxazole (80.8%). Among the isolates tested, 280 (91.0%) were multidrug resistant (resistant to > or =2 agents). The most common resistance pattern was the combination of ampicillin, tetracycline, and trimethoprim/sulfamethoxazole (70.8%). Resistance to ampicillin and tetracycline were more common among S. flexneri than among S. sonnei isolates. CONCLUSIONS: S. flexneri is predominant in Anhui, China, and its higher antimicrobial resistance rate compared with that of S. sonnei is a cause for concern. Continuous monitoring of resistance patterns is necessary to control the spread of resistance in Shigella. The recommendations for antimicrobial treatment must be updated regularly based on surveillance results.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Ampicillin/pharmacology ; Anti-Infective Agents/*pharmacology ; Child ; Child, Preschool ; China ; Drug Resistance, Bacterial/drug effects ; Dysentery, Bacillary/*diagnosis/microbiology ; Feces/microbiology ; Humans ; Infant ; Microbial Sensitivity Tests ; Middle Aged ; Nalidixic Acid/pharmacology ; Shigella/*drug effects/isolation & purification ; Shigella flexneri/drug effects/isolation & purification ; Shigella sonnei/drug effects/isolation & purification ; Tetracycline/pharmacology ; Time Factors ; Trimethoprim-Sulfamethoxazole Combination/pharmacology ; Young Adult

No abstract available.

Polymerase chain reaction (PCR) is a simple, quick and highly sensitive method. However the accuracy of the conventional PCR assay was often affected by false positives and false negatives. In this study, a protocol competitive PCR was used to reduce the false results in screening for selectable marker-free (SMF) Xa2l transgenic rice plants. The competitive template of Xa21 was the endogenous Xa2l homologous sequence located on chromosome 11. The competitive template of the selectable marker gene, hygromycin phosphotransferase (hpt), was an additive DNA extracted from hpt transgenic Nipponbare (Oryza sativa L). Through competitive PCR analysis of transgenic T1 plants produced by double right border binary vector, false positive or false negative samples were effectively diminished, and genuine SMF Xa21 transgenic plants were obviously obtained. Comparing with the conventional non-competitive PCR, competitive PCR increased the accuracy for selecting SMF Xa21 transgenic plants. The results of bacterial blight (BB) resistance tests and hygromycin B resistance assay of SMF Xa21 transgenic plants testified the reliability of this method.
Genetic Vectors ; Oryza ; genetics ; metabolism ; Plant Diseases ; genetics ; prevention & control ; Plant Proteins ; genetics ; metabolism ; Plants, Genetically Modified ; genetics ; metabolism ; Polymerase Chain Reaction ; methods ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Transformation, Genetic

Intestinal ischemia-reperfusion (I/R) is an important event in the pathogenesis of multiple organ dysfunction syndrome (MODS). The aim of this study is to determine the effects of ginsenoside Rb1 on liver injury induced by intestinal I/R in rats. Adult male Wistar rats were randomly divided into four groups: (1) a control, sham-operated group (sham group); (2) an intestinal I/R group subjected to 1 h intestinal ischemia and 2 h reperfusion (I/R group); (3) a group treated with 20 mg/kg ginsenoside Rb1 before reperfusion (Rb1-20 group); and (4) a group treated with 40 mg/kg ginsenoside Rb1 before reperfusion (Rb1-40 group). Liver and intestinal histology was observed. Aspartate aminotransferase (AST), alanine aminotransferase (ALT) level in serum and malondialdehyde (MDA) level in intestinal tissues were measured. Myeloperoxidase (MPO), TNF-alpha, MDA level and immunohistochemical expression of NF-kappa B and intracellular adhesion molecale-1 (ICAM-1) in liver tissues was assayed. In addition, a western blot analysis of liver NF-kappa B expression was performed. Results indicated intestinal I/R induced intestinal and liver injury, which was characterized by increase of AST and ALT in serum, MDA level in intestine, MPO, TNF-alpha and MDA level and ICAM-1 and NF-kappa B expression in the liver tissues. Ginsenoside Rb1 (20, 40 mg/kg) ameliorated liver injury, decreased MPO, TNF-alpha and MDA level, NF-kappa B and ICAM-1 expression in liver tissues. In conclusion, ginsenoside Rb1 ablated liver injury induced by intestinal I/R by inhibiting NF-kappaB activation.
Alanine Transaminase/blood ; Animals ; Aspartate Aminotransferases/blood ; Biological Markers/metabolism ; Ginsenosides/*pharmacology ; Intercellular Adhesion Molecule-1/metabolism ; Intestines/*blood supply/metabolism/pathology ; Ischemia/metabolism/pathology ; Liver/enzymology/pathology ; Liver Diseases/etiology/*pathology ; Male ; Malondialdehyde/metabolism ; NF-kappa B/*antagonists & inhibitors/metabolism ; Peroxidase/metabolism ; Rats ; Rats, Wistar ; Reperfusion Injury/complications/*pathology ; Tumor Necrosis Factor-alpha/metabolism