2.Metabolic shift of Corynebacterium acetoacidophilum-deltaldh under oxygen deprivation conditions.
Qian YANG ; Pu ZHENG ; Fang YU ; Wei LIU ; Zhihao SUN
Chinese Journal of Biotechnology 2014;30(3):435-444
Lactate and succinate were produced by Corynebacterium acetoacidophilum from glucose under oxygen deprivation conditions. To construct knockout mutant, lactate dehydrogenase gene (ldh) of C. acetoacidophilum was deleted by double-crossover chromosome replacement with sacB gene. Comparing with the wild strain ATCC13870, ldhA-deficent mutant produced no lactate with glucose consumption rate decreased by 29.3%, while succinate and acetate concentrations were increased by 45.6% and 182%, respectively. Moreover, the NADH/NAD+ rate was less than 1 (about 0.7), and the activities of phosphoenolpyruvate carboxylase and acetate kinase of the ldhA-deficent mutant were enhanced by 84% and 12 times, respectively. Our studies show that succinicate and acetate production pathways are strengthened by blocking lactate synthesis. It also suggests that improving NADH supply and eliminating acetate generation are alternative strategies to get high succinate-producer.
Corynebacterium glutamicum
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genetics
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metabolism
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Glucose
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Industrial Microbiology
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L-Lactate Dehydrogenase
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genetics
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metabolism
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Lactic Acid
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metabolism
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Oxygen
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metabolism
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Phosphoenolpyruvate Carboxylase
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Succinic Acid
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metabolism
3.Therapeutic effect and immunologic regulation of human amniotic mesenchymal stem cells in rats with experimental type 1 diabetes mellitus
Yingfeng PU ; Ning FANG ; Daixiong CHEN ; Limei YU ; Tao ZHANG ; Chunhua ZHAO
Chinese Journal of Endocrinology and Metabolism 2014;30(1):55-60
Objective To study the therapeutic effect and immunologic regulation of human amniotic mesenchymal stem cells (hAMSCs) in rats with experimental type 1 diabetes mellitus (T1DM).Methods The hAMSCs from human amnion were isolated and cultured in vitro,then phenotype was analyzed by flow cytometry.T1DM were produced by administering streptozocin to rats.The rats were divided into normal control group (n =6),T1 DM model group (n =6),medium treated group (n =6),hAMSCs transplanted group(n =6),and insulin treated group(n =6).5 × 106of hAMSCs or vehicle were administered to rats via sublingual vein.Blood glucose levels of rats were recorded weekly in the groups for six weeks by Blood Sugar Meter.At the end of 6 weeks after hAMSCs transplantation,concentrations of plasma insulin were detected by ELISA; histopathological changes of pancreas,surviwl and differentiation of transplanted hAMSCs in pancreatic tissue were studied with HE staining,and immunofluorescence staining; percentages of lymphocyte subsets in peripheral blood mononuclear cells were determined by flow cytometry ; concentrations of plasma cytokines were determined by cytometric bead array.Results After hAMSCs transplantation,blood glucose levels in rats with T1DM were decreased (P < 0.01),while concentrations of plasma insulin were increased significantly (P<0.01).At 6 weeks,cell-treated animals showed an improvement in pancreas damage ; the percentages of CD4 + IFN-γ+ (Th 1) and C D4 + interleukin (IL)-17 + (Th 17) cells were reduced (all P<0.05),while the percentages of FoxP3-positive regulatory T cells (FoxP3 +Treg) and CD4+ IL-4+(Th2) cells were increased (all P<0.01) ; plasma concentrations of interferon-γ,IL-2,and tumor necrosis factor-αwere decreased (all P<0.01),but IL-4 level was increased (P<0.05).No histological evidence of insulin producing cells from hAMSCs was seen within pancreas.Conclusions hAMSCs may reduce blood glucose and alleviate the islet damage in rats with T1 DM,which is related to their potential to up-regulate FoxP3 +Treg cells.
4.Determination of minimal erythema dose of ultraviolet in 127 volunteers from Urumqi region
Yuan DING ; Fang XIANG ; Shirong YU ; Junqin LIANG ; Xiaojing KANG ; Xiongming PU
Chinese Journal of Dermatology 2012;(12):901-902
Objective To determine the normal range of minimal erythema dose (MED) for ultraviolet A (UVA) and B (UVB) in volunteers from Urumqi region.Methods One hundred and twenty-seven volunteers including healthy subjects and patients with noninflammatory skin disorders were enrolled in this study.SUV-1000 type UV simulator was used as the light source to determine MED of UVA and UVB in these subjects.Results These subjects included 48 persons with Fitzpatrick skin type Ⅲ,79 with Fitzpatrick skin type Ⅳ,51 males and 76 females.The median MED value for UVA and UVB was 38.1 J/cm2 and 31.8 mJ/cm2 respectively in subjects with skin type Ⅲ,59.16 J/cm2 and 48.00 mJ/cm2 respectively in subjects with skin type Ⅳ.Significantly lower median MED values of UVA (both P < 0.01) and UVB (both P < 0.05) were observed in the male and female subjects with skin type Ⅲ compared with those with skin type Ⅳ.The male subjects showed a significantly higher median UVA-MED value (59.16 J/cm2 vs.41.10 J/cm2,P < 0.05),but a similar UVB-MED value (39.60 mJ/cm2 vs.35.55 mJ/cm2,P > 0.05) compared with the female subjects.No significant difference was observed in the median value of UVA-or UVB-MED in subjects with skin type Ⅲ or Ⅳ between Han and Uygur nationality (all P > 0.05).Also,no correlation was found in the median value of UVA-or UVB-MED with age or duration of outdoor exposure in the male or female subjects (all P > 0.05).The lower reference limit was 33.38 J/cm2 for UVA-MED and 27.90 mJ/cm2 for UVB-MED in the population in Urumqi region.Conclusion Skin phototype may be an important determinant of MED.
5.Clinical and MRI features of posterior reversible encephalopathy syndrome in patients with systemic lupus erythematosus
Wei PU ; Hui YU ; Xuming FANG ; Lan CHU ; Shangfu NIU ; Guiquan SHEN
Chinese Journal of Neurology 2015;48(9):753-756
Objective To investigate the clinical and MRI features and pathogenic mechanism of posterior reversible encephalopathy syndrome (PRES) in patients with systemic lupus erythematosus (SLE).Methods Six cases of PRES in SLE proved by integrated clinical diagnosis were collected from January 2008 to December 2013.The clinical and MRI features of these patients were studied retrospectively and the related literatures were reviewed.Results The initial episode of nervous system was involved in 4 cases.The clinical presentations were headache (2 cases),seizures (4 cases),acute confusion state (2 cases),altered mentation (1 case) and vision change (1 case).All cases had lupus nephritis and hypertension,in which 4 cases had renal failure;6 cases were treated with immunosuppressive agents and 4 cases with cyclophosphamide for lupus nephritis when they developed PRES.The vasogenic edema lesions were distributed in the parietal or occipital lobe (5 cases),the frontal lobe (4 cases),temporal lobe (4 cases),basal ganglia (3 cases),splenium (1 case) and cerebellar hemispheres (2 cases).Three major patterns of PRES included dominant parietal-occipital (2 cases),the holohemispheric watershed (3 cases),and superior frontal sulcal (1 case).Conclusions The clinical and MRI findings of PRES in patients with SLE are typical.Awareness of this entity as early as possible can help to improve curative effect and prognosis.
6.A analysis of the risk factors of colorectal polyp malignant transformation
Xiaona YU ; Haoxiang ZHANG ; Pu WANG ; Jingwen LI ; Anran ZHANG ; Yin XU ; Guiyong PENG ; Dianchun FANG
Chongqing Medicine 2014;(29):3866-3869
Objective Colorectal polyp is a precancerous lesion of colorectal cancer .Aim of the study was to explore the risk fac-tors of colorectal polyp malignant transformation .Methods The related information of 75 084 colonoscopies performed from 2003 to 2012 in Southwest Hospital in Chongqing were collected and the relationship between polyp malignant transformation and the pa-tient age ,sex ,polyp location ,size or histological types was analyzed .Results From 2003 to 2012 ,polyps were diagnosed in 14 806 cases of the total 75 084 patients with a 19 .72% detection rate .There were significant difference of the left-side and right-side pol-yp detection rate in different age groups ,and the frequency of polyps distributed in the whole colorectum increased with the increase of age .The rates of epithelial neoplasia and malignant transformation increased with age .At the same time ,malignant transforma-tion rate was significant higher in polyps located in left-side than that in right-side (P<0 .0167) ,in adenoma than that in inflamma-tory hyperplastic polyp (P<0 .01) .The larger diameter and the more villus ,the higher rate of malignant transformation .Conclusion Patient age ,polyp size ,location and histological type could be considered as the significant predictors of colorectal polyp malignant transformation .It may be useful to treat the polyp with endoscopy in patient with age more than 45 and adenoma whose diameter was not less than 1 cm ,located in left-side for prevention of colorectal cancer .
7.Smoking and subjective life qualities in middle school students.
Fang-biao TAO ; Kun HUANG ; Ming GAO ; Pu-yu SU
Chinese Journal of Epidemiology 2006;27(2):132-136
OBJECTIVETo understand the tendency of smoking in middle school students during the last 5 years and to identify the relationship between subjective life qualities and different smoking behaviors in order to determine the possible effects on tobacco use.
METHODSSix middle schools were chosen in the city and rural areas of Hefei in 1998 and 2003, respectively. Smoking behaviors such as ever smoking, smoking before 13 years old, current smoking, regular smoking and addictive smoking were surveyed. Emotional symptoms, satisfaction on school life and on general life were chosen to estimate subjective life qualities. Relationship between subjective life qualities and smoking behaviors was analyzed.
RESULTSPrevalence rates of smoking before 13 years old, current smoking, regular smoking and addictive smoking in 2003 (8.8%, 12.0%, 5.6% and 1.7%, respectively) were higher in 1998 (3.1%, 3.0%, 1.7% and 0.6%, respectively). The prevalence rates of depression and anxiety were high among students with ever smoking, current smoking, regular smoking, addictive smoking, smoking before 13 years old and no attempt to quit cigarette smoking. Lower school life satisfaction and general estimation of life were found among these students. Higher satisfaction on school life (OR = 0.657) and on ordinary life (OR = 0.766) were protecting factors of current smoking but comorbid depression and anxiety were risk factors (OR = 2.181). Higher school life satisfaction (OR = 0.388) seemed to be protecting factor of addictive smoking while depression was risk factor of addictive smoking (OR = 2.753) and regular smoking (OR = 1.676).
CONCLUSIONSmoking behaviors were common in middle school students and affected by several emotional and cognitive factors of subjective life qualities. Longitudinal designs are required to clarify the causal relationship between these factors and smoking behaviors in adolescents.
Adolescent ; Adolescent Behavior ; Anxiety ; epidemiology ; China ; epidemiology ; Depression ; epidemiology ; Health Surveys ; Humans ; Prevalence ; Psychology, Adolescent ; Quality of Life ; Risk Factors ; Rural Population ; statistics & numerical data ; Schools ; Smoking ; epidemiology ; Students ; psychology ; statistics & numerical data
8.Analysis of BRAF gene mutations in 80 patients with malignant melanoma in Xinjiang Uygur Autonomous Region
Fang GUO ; Xiaojing KANG ; Xiaohui TANG ; Zhenzhu SUN ; Xiongming PU ; Jing LI ; Wenjing CHEN ; Ying JIN ; Dezhi ZHANG ; Shirong YU
Chinese Journal of Dermatology 2013;(1):33-36
Objective To assess the relationship between BRAF gene mutations and clinical phenotype of malignant melanoma.Methods Tissue specimens were collected from the lesions of 80 patients with malignant melanoma,and from the normal skin of 30 patients with trauma in the Department of Plastic Surgery or General Surgery,and subjected to paraffin embedding and DNA extraction.PCR was performed to amplify the exon 11 and 15 of BRAF gene followed by DNA sequencing.Chi-square test and Fisher's exact test were carried out to assess the relationship between BRAF gene mutations and clinical phenotypes of malignant melanoma.Results BRAF gene mutations were found in 19 (23.8%) of the 80 malignant melanoma specimens.Among the 19 mutationpositive specimens,17 (88.2%) carried mutations in exon 15 of BRAF gene with V600E as the most frequent (88.2%,15/17) mutation type,and 2 (10.5%) carried mutations in exon 11.No mutation was found in any of the normal skin tissue specimens.The average age at onset was 57.5 years in these patients.The frequency of BRAF gene mutation was significantly higher in patients younger than 60 years than in those older than 60 years (37.1% vs.13.3%,x2=6.613,P < 0.05).A significant difference was observed in the frequency of BRAF gene mutation among tissue specimens of mueosal,acral and non-aeral malignant melanoma (18.2% (4/21) vs.14.7%(5/34) vs.41.7% (10/24),x2=6.167,P < 0.05).There was no significant association between BRAF gene mutation and gender,race or lymph node metastasis (all P > 0.05).Conclusions BRAF gene is a hot spot for mutations in patients with malignant melanoma in Xinjiang Uygur Autonomous Region,with V600E point mutation in exon 15 as the most frequent mutation type.BRAF gene mutations appear to be closely correlated with the age at onset of and lesional sites in,but uncorrelated with gender and race of or lymph node metastasis in,patients with malignant melanoma.
9.An investigation on self-harm episodes and their relationship with suicidal psychology and behaviors in 2713 college students
Pu-Yu SU ; Jia-Hu HAO ; Zhao-Hui HUANG ; Fang-Biao TAO
Chinese Journal of Epidemiology 2010;31(11):1267-1271
Objective To investigate the episodes and influencing factors on self-harm and to explore the relationship between self-harm episodes and suicidal psychology and behaviors in college students. Methods Four universities were selected using cluster sampling method in Anqing city and Chaohu city. Totally, 2713 college students completed this survey. Data were analyzed by Pearson Chi-square and logistic regression. Results In the last six months, rates of highly lethal self-harm,less lethal self-harm with visible tissue damage, self-injury without visible tissue damage,self-harmful behaviors with latency damage, other self-harmful behaviors with menticide were 1.9%,5.5%, 15.3%, 21.2% and 17.0% respectively. The total rate of self-harm was 31.3%. 73.1% of the students with self harmful experiences had the above mentioned behaviors more than 3 times in the last six months. The top 3 reasons for taking self-harm actions were: having learning problems (43. 1% ), failed love affairs (25.0%) and having conflicts with others (23.9%). There were different influencing factors among different kinds of self-harm episodes. Depression was the risk factor of self-harm. The higher score of having high self-esteem was the protective factor of all kinds of self-harm actions except highly lethal ones. Higher score of difficulties in identifying feelings was one of the risk factors. The rates of suicidal psychology and behaviors in students with self-harm were significantly higher than those in students without those behaviors. Result from linear x2 test indicated that the graveness of tissue damage of self-harm was higher along with the rates of suicidal psychology and behaviors (P<0.01). Conclusion Among 2713 college students, about 1/3adolescents having experienced self-harm in the last 6 months, many with repeated ones. Depression and difficulties in identifying feelings were the two risk factors while self-esteem was the protective factor related to most of the self-harm cases.
10.Fusion Expression of Cecropin X Including the Cleavage of FXa in Escherichia coli
Liu-Di YUAN ; Fei DOU ; Yu-Pu LIANG ; Wei XIE ; Fang WANG ; Shuang-Quan ZHANG ; Zhu-Ying DAI
Chinese Journal of Biotechnology 2000;16(3):411-414
PCR method was used to introduce the code sequence of Factor Xa cleavage site to the 5′ end of cecropin CMIV mutant gene X, then the gene was cloned into the expression vector pGEX-KG, and was highly expressed in E. coli BL21 by IPTG induction. The fusion protein was purified by affinity-chromatography and was cleaved by Factor Xa. Cecropin X with antibacterial activity was obtained after purified by ion-exchange chromatography.