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Sichuan Da Xue Xue Bao Yi Xue Ban ; 50(5): 714-719, 2019 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-31762243


OBJECTIVE: To select and identify the bacterium which highly produces protease and ß-D-glucosidase from 72 strains of Shuidouchi from Sichuan, and to provide evidence for further research on its nutritional value and fermentation strain exploiting. METHODS: Casein degradation test and pNPG chemical test were applied respectively to detect the capacity to produce protease and ß-D-glucosidase of each strain. Characteristics of morphology, biochemistry, 16S rRNA and MALDI-TOF-MS were used to identify the fermentation strain, which genetic stability, curves of growth and enzyme producing were also obtained. RESULTS: The strain with the highest enzyme activity of ß-D-glucosidase (0.084 U/L) among the top 10 strains for producing protease was selected as the fermentation strain and was identified as Bacillus subtilis, which curves of growth and enzyme producing conformed as well. The result of genetic stability showed that capacity of enzyme producing was stable until the 10th generation. CONCLUSIONS: The fermentation strain which highly produced protease and ß-D-glucosidase was selected from 72 strains of shuidouchi from Sichuan and was identified as Bacillus subtilis.

Bacillus subtilis/enzimologia , Glucosidases/biossíntese , Peptídeo Hidrolases/biossíntese , Alimentos de Soja/microbiologia , China , Fermentação , RNA Ribossômico 16S
Infect Genet Evol ; 67: 44-50, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30347249


Enterovirus 71 is the main pathogen that causes severe and fatal hand-foot-mouth-disease (HFMD) cases. As the enterovirus virus mutation has implications for pathogenesis, vaccine development, antiviral therapy, and epidemiological disease management of the virus. In this study, we investigated the variations of enterovirus 71 in thermal adaption, using the method of adaptive laboratory evolution. The sibling virus strains were isolated from a 2-year-old severe case of HFMD (#100) and her symptomless close contact (#101). Both strains were cultured in Vero cells by serial passage of 36 generations at the temperatures of 28.0 °C, 33.0 °C and 39.5 °C to construct adaptive lineages. According to the comparative analysis of phenotypes between adapted strains and parental strains, differences in growth rate were observed in the sibling lineages and a larger plaque was found mainly in the hot adapted strains for lineage #101. Two sets of adaptive strains from six time points (parental, 12th 17th, 31st, 35th passage and endpoint) were sequenced and analyzed by both Sanger sequencing and Next Generation Sequencing. Several variations in most coding genes and one reverse mutation in 5'UTR was observed, along with the identity of 99.8% for complete genome for both lineages. Notably, thermal specific non-synonymous mutations were found in the gene of VP1\VP3\3A\2C\3C. Moreover, the concurrent mutations A292G, A434G and A355C/T of sibling lineages in VP1 showed quantificational trace with distinguishing patterns for different temperatures, which were suspected to be the thermo-sensitive mutation hotspots. These results highlight the possible rules of thermal adaption in enterovirus 71, produce a novel picture of genome evolution of the virus, and shed light on viral variation and evolution.

Adaptação Biológica , Enterovirus Humano A/classificação , Enterovirus Humano A/genética , Temperatura Ambiente , Regiões 5' não Traduzidas , Animais , Evolução Biológica , Células Cultivadas , Infecções por Enterovirus/virologia , Genoma Viral , Humanos , Análise de Sequência de DNA , Células Vero , Ensaio de Placa Viral , Replicação Viral
Chin J Integr Med ; 25(6): 425-430, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29790063


OBJECTIVE: To examine the effect of the aqueous extract of Ligustrum robustum on tumor growth in vitro and in vivo and explore the possible molecular mechanisms. METHODS: In in vitro study, cell viabilities of human cervical carcinoma cells (HeLa), human breast cancer cells (MCF-7), human prostate cancer cells (PC-3), human hepatoma cells (7721) and human colon carcinoma cells (SW480) were evaluated with cell counting kit-8. For L. robustum-treated Hela cells, early or late apoptosis were evaluated by annexin V/PI staining. Mitochondrial membrane potential was measured by staining cells with JC-1. Apoptosis was monitored by nuclear morphology based on chromatin condensation and fragmentation by 4',6-diamidino-2-phenylinole (DAPI) staining. Caspase-3 and -8 activity levels were measured by a colorimetric assay. In vivo, to evaluate the possible mechanism of L. robustum-mediated antitumor effect, nude mouse xenograft study was also conducted. RESULTS: In in vitro study, L. robustum was found to be toxic to HeLa, MCF-7, PC-3, 7721, SW480, with an half maximal inhibitory concentration value of 2-5 mg/mL (P<0.05). Moreover, externalization of phosphatidylserine, loss of mitochondrial membrane potential, DNA fragmentation and activation of caspase-3 and -8 were detected in L. robustum-treated Hela cells. Using a nude mouse model bearing Hela xenografts, we found that L. robustum reduced tumor volume and tumor weight (P<0.05), but had no effect on body weight and histological damage of important organs. Intraperitoneal injection of L. robustum caused a significant reduction in serum aspartate transaminase and alanine transaminase levels (P<0.05). Furthermore, cleaved caspase-3-positive and terminal nucleotidyl transferase-mediated nick end labeling (TUNEL)-positive cells were observed in L. robustum-treated tumor tissues. CONCLUSIONS: L. robustum inhibits tumor cell growth both in vitro and in vivo by inducing apoptosis in a caspase-dependent way without apparent hepatic toxicity and histological damage, which may offer partial scientific support for the ethnopharmacological claims of L. robustum as a herbal tea for its antitumor activity.

Antineoplásicos/farmacologia , Ligustrum/química , Chás de Ervas , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Feminino , Humanos , Camundongos Nus , Carga Tumoral/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
World J Gastroenterol ; 21(46): 13042-54, 2015 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-26676281


AIM: To investigate the anti-obesity and antibacterial effects of Ligustrum robustum (L. robustum) in vivo and in vitro and its possible mechanisms. METHODS: The effects of L. robustum aqueous extract (LR) on various gut bacteria in vitro were evaluated. The effects of LR on high-fat diet-fed (HFD) rats in vivo were also assessed. Culture methods, quantitative polymerase chain reaction, and terminal-restriction fragment length polymorphism were used to analyze the effects of LR on gut bacteria. Biochemical tests were also performed to detect the changes in obesity-related indicators after LR treatment. RESULTS: LR treatment lowered adipose weight and decreased Lee's index, blood glucose, total cholesterol, and lipid in the tested groups relative to control (P < 0.05). To determine the reasons for these changes, we assessed the potential bacteriostatic and bactericidal effects of LR on specific bacterial species in vitro. LR affected the richness, diversity, and evenness of gut bacteria, increased fecal Lactobacillus, and decreased Enterococci in HFD rats (P < 0.05). CONCLUSION: L. robustum may be a safe and effective food for weight loss and obesity control, and the effects of L. robustum might be mediated by the regulation of gut bacteria.

Antibacterianos/farmacologia , Fármacos Antiobesidade/farmacologia , Bactérias/efeitos dos fármacos , Microbioma Gastrointestinal/efeitos dos fármacos , Intestinos/efeitos dos fármacos , Ligustrum/química , Obesidade/prevenção & controle , Extratos Vegetais/farmacologia , Adiposidade/efeitos dos fármacos , Animais , Antibacterianos/isolamento & purificação , Fármacos Antiobesidade/isolamento & purificação , Bactérias/classificação , Bactérias/crescimento & desenvolvimento , Dieta Hiperlipídica , Modelos Animais de Doenças , Intestinos/microbiologia , Masculino , Obesidade/microbiologia , Obesidade/fisiopatologia , Fitoterapia , Extratos Vegetais/isolamento & purificação , Plantas Medicinais , Ratos Sprague-Dawley , Perda de Peso/efeitos dos fármacos
World J Gastroenterol ; 17(8): 1076-81, 2011 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-21448362


AIM: To investigate the differences in cultivable gut bacteria and peroxisome proliferator-activated receptor γ2 (PPAR-γ2) gene Pro12Ala variation in obese and normal-weight Chinese people. METHODS: Using culture methods, the amounts of Escherichia coli, Enterococci, Bacteroides, Lactobacilli, Bifidobacteria and Clostridium perfringens (C. perfringens) in the feces of 52 obese participants [body mass index (BMI): ≥ 28 kg/m(2)] and 52 participants of normal-weight (BMI: 18.5-24 kg/m(2)) were obtained. Study participants completed comprehensive questionnaires and underwent clinical laboratory tests. The polymerase chain reaction-restriction fragment length polymorphism (PCR-PFLP) assay was used to analyze PPAR-γ2 gene Pro12Ala variation. RESULTS: The obese group exhibited a lower amount of C. perfringens (6.54 ± 0.65 vs 6.94 ± 0.57, P = 0.001) and Bacteroides (9.81 ± 0.58 vs 10.06 ± 0.39, P = 0.012) than their normal-weight counterparts. No major differences were observed in Pro12Ala genotype distribution between the two groups; however, obese individuals with a Pro/Ala genotype had a significantly lower level of Bacteroides (9.45 ± 0.62 vs 9.93 ± 0.51, P = 0.027) than those with a Pro/Pro genotype. In addition, the obese group demonstrated a higher stool frequency (U = 975, P < 0.001) and a looser stool (U = 1062, P = 0.015) than the normal-weight group. CONCLUSION: Our results indicated interactions among cultivable gut flora, host genetic factors and obese phenotype and this might be helpful for obesity prevention.

Bactérias/metabolismo , Trato Gastrointestinal/microbiologia , Obesidade/genética , Obesidade/fisiopatologia , Polimorfismo Genético , Peso Corporal/genética , Feminino , Genótipo , Humanos , Masculino , PPAR gama/genética , Inquéritos e Questionários
Sichuan Da Xue Xue Bao Yi Xue Ban ; 40(5): 865-8, 876, 2009 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-19950601


OBJECTIVE: To study on the polymorphism of UCP2 gene in Chengdu simple obesity and normal-weight people and to initially investigate the relationship between UCP2 Ala55Val variation and gut bacteria. METHODS: PCR-PFLP was applied to determine the genotypes of Ala55Val variant in the UCP2 gene of 86 Chengdu people (the simple obesity group, 43 subjects; the normal-weight group, 43 subjects). And six kinds of gut bacteria among different genotypes in different groups were analyzed. RESULTS: Both the simple obesity and the normal-weight group had the Ala55Val variants of Ala/Ala, Val/Val and Ala/Val in the UCP2 gene, and the Ala55Val genotype distributions between the two groups was significantly different (chi2=11.97, P< 0.05). The allelic mutation frequency in the simple obesity group was higher than that of the normal-weight group (chi2=10.06, P<0.05). However, no significant difference was observed in the population of six gut bacteria among different genotypes in different groups (P>0.05). CONCLUSION: The UCP2 gene mutation might be a risk factor of obesity in Chengdu area. However, this gene mutation may not be an impact factor on the alternation of gut bacteria.

Intestinos/microbiologia , Canais Iônicos/genética , Proteínas Mitocondriais/genética , Mutação , Obesidade/genética , Polimorfismo Genético , Adolescente , Adulto , Alelos , China , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Proteína Desacopladora 2 , Adulto Jovem