By using enrichment culture in liquid minimal medium or direct culture on minimal medium plates, thirteen bacterial strains (AD27-AD39) capable of utilizing atrazine as a sole nitrogen source for growth were isolated from a mixture of industrial wastewater and sludge from an atrazine manufacturing plant. Based on 16S rRNA gene sequencing, eleven strains were identified as Arthrobacter spp. and two strans were identified as Pseudomonas spp.. We further studied in detail the composition of atrazine-degrading genes and degradation characteristics of Arthrobacter sp. AD30 and Pseudomonas sp. AD39 that have high degradative activity. From PCR assays, it was indicated that both AD30 and AD39 strains contained atrazine-degrading genes trzN and atzBC and was capable of degrading toxic atrazine to nontoxic cyanuric acid. The biodegradation experiments showed that the percentage of atrazine removal were 92.5%,were 92.5%, 97.9% and 99.6% respectively after AD30, AD39 or the mixture of the two strains were inocu- lated and incubated at 30?C for 48 hours in minimal media containing 200 mg/L atrazine, indicating that atrazine degradation by the mixed bacteria was more effective than the single strain. In addition, after industrial wastewater containing 176 mg/L atrazine was inoculated with the mixed bacteria and incubated at 30?C with shaking for 72 hours atrazine were removed by 99.1%, implicating that the mixed bacteria are good candidate for biotreatment of atrazine-containing industrial wastewater.

Adult ; Aged ; Bursitis ; therapy ; Exercise Therapy ; methods ; Female ; High-Energy Shock Waves ; Humans ; Male ; Middle Aged ; Treatment Outcome

Objective To investigate the expression of transforming growth factor β1,transforming growth factor beta receptor(TBR)Ⅰ,TβR Ⅱ,Smad4 and C-Jun in rats with nonalcoholic fatty liver disease(NAFLD)and to find out the mechanisms of liver fibrosis in patients with NAFLD.Methods A total of 18 male SD rats were randomly divided into normal control group(n=9)and model group(n=9).The rats in control group were fed with normal diet,and those in model group were fed with fat-rich diet(consisted of 10%lard oil+2%cholesterol).An rats were sacrificed at the 20th week.The levels of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were examined by RT-PCR.The expressions of TGFβ1 and Smad4 in liver tissue were detected by immunohistochemistry.The expression of C-Jun protein was detected by Western blotting.Results The NAFLD model was successfully established.The immunohistochemistry examination revealed that TGFβ1 and Smad4 were expressed weekly in control group,but strongly expressed in model group.RT-PCR showed that A values of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were 0.46±0.12,5.z4±2.70 and 3.35±1.95,respectively,in model group,which were higher than those in control group(0.21±0.09,1.36±0.77 and 0.52±0.19,all P values＜0.01).The Western blotting results demonstrated that the expression of C-Jun protein in model group(0.93±0.41)was higher than that in control group (0.32±0.25,P=0.001).Conclusion TGFβ1/Smad4 signal pathway might be involved in the development of hepatic fibrosis in NAFLD.Blocking TGFβ1/Smad4 signal pathway will be helpful in treatment of NAFLD.

BACKGROUNDOverexpression of G-protein coupled receptor 34 (GPR34) affects the progression and prognosis of human gastric adenocarcinoma, however, the role of GPR34 in gastric cancer development and progression has not been well-determined. The current study aimed to investigate the effect of GPR34 knockdown on the proliferation, migration, and apoptosis of HGC-27 gastric cancer cells and the underlying mechanisms.

METHODSThe expression of GPR34 in gastric cancer cell line HGC-27 was detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. HGC-27 cells were employed to construct the stable GPR34 knockdown cell model in this study. Real-time RT-PCR and Western blotting were applied to validate the effect of short hairpin RNA (ShRNA) on the expression of GPR34 in HGC-27 gastric cells. The proliferation, migration of these cells were examined by Cell Counting Kit-8 and transwell. We also measured expression profile of PI3K/PDK1/AKT and ERK using Western blotting.

RESULTSThe ShRNA directed against GPR34 effectively inhibited both endogenous mRNA and protein expression levels of GPR34, and significantly down-regulated the expression of PIK3CB (P < 0.01), PIK3CD (P < 0.01), PDK1 (P < 0.01), phosphorylation of PDK1 (P < 0.01), Akt (P < 0.01), and ERK (P < 0.01). Furthermore, GPR34 knockdown resulted in an obvious reduction in HGC-27 cancer cell proliferation and migration activity (P < 0.01).

CONCLUSIONSGPR34 knockdown impairs the proliferation and migration of HGC-27 gastric cancer cells in vitro and provides a potential implication for therapy of gastric cancer.

Apoptosis ; genetics ; physiology ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; genetics ; physiology ; Humans ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Receptors, Lysophospholipid ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism

Carcinoma, Hepatocellular ; pathology ; Humans ; Liver Neoplasms ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Peptide Fragments ; pharmacology ; Peptides ; pharmacology ; Tumor Cells, Cultured

Adult ; Aldosterone ; blood ; Angiotensin II ; blood ; Diet, Sodium-Restricted ; Female ; Humans ; Kidney ; physiopathology ; Kidney Function Tests ; Liver Cirrhosis ; blood ; physiopathology ; Male ; Middle Aged ; Renin ; blood ; Sodium ; administration & dosage

To block tumor cell adhesion, inhibit tumor metastasis and recurrence, the anti-adhesion peptide-trimeric beta peptide (DLYYLMDLSYSMKGGDLYYLMDLSYSMKGGDLYYLMDLSYSMK, beta3) was designed. The DNA fragment of beta3 was cloned into expression vector pET-His and the fusion protein His-beta3 was expressed in E. coli. BL21(DE3)plysS. After 1.5 hours' induction with IPTG, His-beta3 peptide was expressed significantly amounting to 10% of the insoluble proteins and 4% of the total proteins. 20mg of beta3 peptide was obtained from one litter culture medium after purification by using metal-chelating sepharose 6B FF. The purity of beta3 is 92.2% according to Gel-Pro analysis. The anti-adhesion effects of beta3 peptide, beta1 peptide (DLYYLMDLSYSMK) and GRGDS on the hepatocellular carcinoma cell line SMMC-7721 and the high metastasis hepatocellular carcinoma cell line HCCLM6 were studied. The result showed the beta3 blocked the adhesion of HCCLM6 cells and SMMC-7721 cells to fibronectin (FN) specifically. The inhibition effect was dose-dependent and time-dependent and the inhibition rate of beta3 was higher than three times concentration of beta1 and GRGDS. This suggested that pET-His-beta3/BL21(DE3)plysS was a suitable expression system for beta3, and the expressed beta3 specially inhibited the adhesion of cancer cells.
Amino Acid Sequence ; Antineoplastic Agents ; pharmacology ; Cell Adhesion ; drug effects ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Peptide Fragments ; pharmacology ; Peptides ; genetics ; metabolism ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; Tumor Cells, Cultured

Objective To investigate the possibility of MRI on visualizing the relationship between glossopharyngeal nerve and surrounding vessels,and to evaluate the significance of MRI in the diagnosis and treatment of glossopharyngeal neuralgia.Methods MRI findings were analyzed retrospectively in 12 patients with glossopharyngeal neuralgia,and were compared with surgical findings and effect of pain relief.Results The artery compression or contact of the glossopharyngeal entry zone,as revealed during operation in l0 patients with glossopharyngeal neuralgia,was visualized on MRI in 9 and not seen in 1.The venous compression of the glossopharyngeal entry zone was not identified on MRI in 1.The conglutinative arachnoids of the glossopharyngeal entry zone was not visualized on MRI in 1.MRI demonstrated the affected glossopharyngeal nerve root entry zone was compressed or contacted by the posterior inferior cerebellar artery (PICA)in 8 patients and by the vertebral artery in 1 patient.One patient's offending vessel was confirmed to be the anterior inferior cerebellar artery(AICA)by the operation,and the surgical findings were corresponded with MRI in others.Vascular compression or contact of the affected glossopharyngeal nerve was not visualized on MRI in 3 patients,and operation confirmed that the glossopharyngeal nerve root entry zone was compressed by unknown artery in 1,by small vein in 1,and by eonglutinative araehnoids in 1, respectively.Eight patients presented with symptoms of the ipsilateral trigeminal neuralgia concurrently.The compression of the affected trigeminal nerve root by superior cerebellar artery(SCA)was visualized on MRI in 6 patients,and operation did not reveal the source of artery compression in 1 and corresponded with MRI findings in other 5 cases.Vascular compression of affected trigeminal nerve was not visualized on MRI in 2 patients,and intraoperative inspection revealed that trigeminal nerve root was compressed by draining vein of brainstem in 1 and not compressed by any vessels in 1.All patient's neuralgia resolved after microvascular decompression of glossopharyngeal nerve and trigeminal nerve.Conclusion It is possible to visualize the glossopharyngeal and surrounding arteries on MRI,and it is of great significance in the diagnosis and treatment of this kind of glossopharyngeal neuralgia.

OBJECTIVETo study the changes of insulin sensitivity in individuals with impaired glucose tolerance and diabetes and the relationship between insulin sensitivity and over weight/obesity (OW/OB) .

METHODSFifty-two individuals were divided into 4 groups according to WHO diagnostic criteria of obesity (1998) and diabetes (1999): normal weight with normal glucose tolerance (NW-NGT) group, OW/OB with normal glucose tolerance (OW/OB-NGT) group, OW/OB with impaired glucose tolerance (OW/OBIGT) group and OW/OB with diabetes mellitus (OW/OB-DM) group. Individuals in OW/OB-NGT group were further classified into 3 subgroups: over weight subgroup, mild obesity subgroup, and mediate obesity subgroup. Abdominal fat area was measured with magnetic resonance imaging. Visceral obesity was defined as intra-abdominal fat area over 100 cm(2). All subjects with NGT were divided into visceral obesity (VA) group and non-visceral obesity ( Non-VA) group. Extended hyperinsulinemic euglycemic clamp was performed to assess the peripheral tissue insulin sensitivity in all subjects.

RESULTSThe rates of insulin mediated glucose disappearance (Rd) were (3. 25+/-0. 13) mg x kg (-1) min (-1) in OW/OB-NGT group, (3. 06+/-0. 26) mg x kg(-1) x min(-1) in OW/OB-IGT group, and (3.19+/-0.44) mg x kg(-1) x min (-1) in OW/OB-DM group, which were significantly lower than that in NW-NGT group [ (5. 86+/-0. 65) mg x kg (-1) min (-1) ] (P < 0. 05, P < 0.01). The Rd in over weight subgroup [(3.50+/-0. 19) mg kg(-1) x min(-1) ] , mild obesity subgroup [(3. 03+/-0. 13) mg x kg (-1) min(-1)] , and mediate obesity subgroup [(2. 75 +/-0. 24) mg x kg (-1) min(-1)] were significantly lower than that of NW-NGT group (P <0. 05, P <0. 01). The Rd [ (2. 97+/-0. 12) mg kg(-1) x min(-1) vs (4.55+/-0.43) mg x kg(-1) x min(-1)] and glucose oxidation [(1.47 +/-0. 19) mg x kg(-1) min(-1) vs (2.24 +/-0. 19) mg kg(-1) x min(-1) in VA group were significantly decreased than that in non-VA group (P < 0. 05, P < 0. 01). Body mass index, waist and hip ratio, waist circumference, and intra-abdominal fat area were negatively correlated with Rd, respectively (P < 0. 01). Multiple regression analysis showed that body mass index, intra-abdominal fat area and abdominal subcutaneous fat area were the main risk factors of insulin sensitivity.

CONCLUSIONSInsulin sensitivity decreased in OW/OB individuals with or without hyperglycemia. Insulin sensitivity is lower in subjects with visceral obesity. Total body fat and abdominal fat are the main risk factors of insulin sensitivity.

Adolescent ; Adult ; Body Mass Index ; Female ; Glucose Clamp Technique ; methods ; Glucose Intolerance ; etiology ; Humans ; Insulin Resistance ; Intra-Abdominal Fat ; metabolism ; Male ; Middle Aged ; Obesity ; complications ; physiopathology ; Subcutaneous Fat, Abdominal ; metabolism

BACKGROUNDOxymatrine has certain antiviral effects in the treatment of chronic hepatitis B (CHB), but its exact mechanism is unclear. The objective of the present study was to explore oxymatrine's antiviral mechanism by studying its effect on the hepatitis B virus (HBV) specific cytotoxic T lymphocyte (CTL) surface programmed death receptor-1 (PD-1) expression in CHB patients.

METHODSSixty-five CHB patients who had HBV DNA(3)10(4) copies/ml, positive HBeAg, positive human leukocyte antigen (HLA)-A2, alanine aminotransferase (ALT) > 2 x upper limit of normal value (ULN) were randomly divided into two groups: treatment group (n = 33), treated with an intravenous infusion of 600 mg oxymatrine in glucose solution once a day for a month, then with a 200 mg oxymatrine oral capsule three times a day, and a 200 mg silibin meglumine tablet three times a day; control group (n = 32) patients were treated only with silibin meglumine tablet, method and dosage were the same as those of treatment group. Three months later, peripheral blood HBV-specific CTL surface PD-1 expression, HBV-specific CTL level, HBV DNA, HBeAg, and results of liver function tests were analyzed and compared.

RESULTSThree months post-treatment, in the treatment group, peripheral blood HBV-specific CTL surface PD-1 expression ((19.42 ± 15.94)%) decreased significantly compared to the pretreatment level ((31.30 ± 24.06)%; P < 0.05), and decreased significantly compared to that of control group three months after treatment ((29.45 ± 21.62)%; P < 0.05). HBV-specific CTL level ((0.42 ± 0.07)%) significantly increased compared with the pretreatment ((0.29 ± 0.15)%; P < 0.01), and the control group posttreatment level was (0.31 ± 0.15)% (P < 0.05). HBV DNA level in 11 cases became negative (HBV DNA < 500 copies/ml, 33.33%), which was higher than that of the control group after treatment (two cases, 6.25%; χ(2) = 7.45, P < 0.01), HBeAg of nine cases turned negative (27.27%), which was higher than that of the control group after treatment (one case, 3.13%; χ(2) = 7.27, P < 0.01).

CONCLUSIONOxymatrine could downregulate peripheral blood HBV-specific CTL surface PD-1 expression in CHB patients, increase HBV-specific CTL level, which may be one of the possible mechanisms by which oxymatrine clears or inhibits HBV in CHB patients.

Adult ; Alkaloids ; therapeutic use ; Antiviral Agents ; therapeutic use ; DNA, Viral ; blood ; Female ; Hepatitis B virus ; immunology ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Male ; Middle Aged ; Programmed Cell Death 1 Receptor ; analysis ; Quinolizines ; therapeutic use ; T-Lymphocytes, Cytotoxic ; chemistry