1.A Rapid and Efficient Method for Directed Screening of Lipase-producing Burkholderia cepacia Complex with Organic Solvent Tolerance from Plant Rhizosphere
Zhengyu SHU ; Ruifeng LIN ; Huan JIANG ; Yanfeng ZHANG ; Jianzhong HUANG
Microbiology 1992;0(06):-
Lipase from Burkholderia cepacia complex is one of the most versatile biocatalyst and is used widely in many biotechnological application fields including detergent additives, the resolution of racemic compounds, etc. Based on the known whole genomic information of B. cepacia, both ampicillin and kanamycin were added to the TB-T media, the traditional selective media, to screen B. cepacia complex strains from rhizosphere soil samples. The single colonies on the plates with the modified TB-T media were then qualitatively determined the ability to produced the extracellular lipase in the rhodamine B-olive oil agar plates. Thirty-five strains of lipolytic pseudo-B. cepacia complex were isolated and the positive rate of lipolytic bacteria was 65%. Among them, 15 pseudo-B. cepacia complex strains with the tolerance to benzene, n-hexane and n-heptane at the concentration of 10% (V/V) were selected and identified by the recA gene sequence. All of the 15 lipolytic bacteria belonged to the B. cepacia complex.
2.The effect of therapeutic life style on serum FFA and constitution index in patients with coronary heart disease complicated metabolic syndrome
Xiaoyong LIU ; Ge ZHANG ; Ruifeng YANG ; Zhi LIN ; Rui YANG
Chongqing Medicine 2015;(22):3099-3101,3104
Objective To investigate the level of serum free fatty acid (FFA )after improving the life style in patients with coronary heart disease complicated metabolic syndrome and the effect of therapeutic life style on traditional risk factors of coronary artery disease.Methods A total of 395 patients with coronary heart disease complicated metabolic syndrome were recruited.Pa-tients were divided into intervention group (group A,conventional drug therapy+ intensive life style intervention,n=97)and non-intervention group (group B,conventional drug therapy,n=38)according to the scores of life style.Serum free fatty acid (FFA) was determined by ELASA.The scores of life style was obtained bylife style questionnaire.Results (1)The serum FFA of pa-tients with coronary heart disease complicated metabolic syndrome were positively related to waist circumference and waist-high-ra-tio.(2)Waist circumference,BMI and FFA of group A were significantly lower than those in group B after therapeutic life style in-tervention(P <0.05).(3)Compared with the baseline,the constitution index and FFA in group A were significantly lower after 6-months therapeutic life style intervention(P <0.05).Conclusion Therapeutic life style can reduce the level of FFA and constitution index of the patients with coronary heart disease complicated metabolic syndrome.
3.Correlations of serum lipoprotein (a) with ischemic stroke and its etiological subtypes
Xiaoqiang LIU ; Weiduan ZHUANG ; Ruifeng WANG ; Lan LIN ; Yingxiu XIAO
International Journal of Cerebrovascular Diseases 2016;24(12):1062-1067
Objective To investigate the correlation between serum lipoprotein (a) (Lp(a)) level andischemic stroke and its etiological subtypes. Methods The consecutive inpatients with acute ischemic stroke (case group) and age-and sex-matched healthy subjects (control group) over the same period were enrolled retrospectively. The demographic and baseline clinical data, as well as fasting blood glucose, fibrinogen,homocysteine, total cholesterol, triacylglycerol, high-densitylipoprotein cholesterol, low -density lipoprotein cholesterol, and Lp(a) concentration of the case group and the control group were collected. According to TOAST classification criteria, the patients in the case group were divided into large artery atherosclerosis (LAA), small artery occlusion (SAO) and cardioembolism (CE), and the patients with other determined etiology and undetermined etiology were excluded. Multivariate logistic regression analysis was used to make clear the correlation between serum Lp(a) and acute ischemic stroke and its etiological subtypes. Results A total of 214 patients with ischemic stroke were enrolled. Ninety-seven had LAA (45.33%), 64 (29.91%) had SAO, and 53 (24.77%) had CE. There were 118 subjects in the control group. There were significant differences in the proportions of hypertension, diabetes, hyperlipidemia, atrial fibrillation and alcohol consumption, as well as systolic blood pressure, diastolic blood pressure, fasting blood glucose, total cholesterol, low -density lipoprotein cholesterol, Lp(a), fibrinogen, and homocysteine between the case group and the control group (all P <0.001). Multivariate logistic regression analysis showed that after adjustment for age and sex, Lp(a) is an independent risk factor for ischemic stroke (odds ratio [OR] 2.014, 95% confidence interval [CI ] 1.273-3.092, P = 0.036). The independent risk factors for LAA included hypertension (OR 3.353, 95% CI 1.714-6.558, P = 0.001), systolic blood pressure ( OR 2.786, 95% CI 1.136-5.538, P =0.016), homocysteine ( OR 1.108, 95% CI 1.031-2.191, P = 0.005), total cholesterol (OR 2.169, 95% CI 1.599-4.943, P = 0.001), low -density lipoprotein cholesterol ( OR2.782, 95% CI 1.093-5.238, P =0.024), and Lp(a) (OR 3.072, 95% CI 1.907-8.064, P =0.001). Theindependent risk factors for SAO included hypertension ( OR 7.042, 95% CI 3.189-25.55, P =0.001), diabetes mellitus (OR 5.162, 95% CI 2.372-11.23, P =0.001), fibrinogen (OR 1.667, 95% CI 1.434-2.025, P = 0.045), and homocysteine (OR 1.967, 95% CI 1.859-1.995, P =0.036). The independent risk factors for CE included atrial fibrillation (OR 13.340, 95% CI 4.637-39.20, P = 0.001), fibrinogen (OR 2.365, 95% CI 1.147- 4.904, P =0.029), and Lp(a) (OR 1.656, 95% CI 1.996-3.001, P = 0.035). Conclusions Lp(a) is an independent risk factor for ischemic stroke, and can be used as a serum biomarker for predicting the risk of the onset of ischemic stroke. There are differences in independent risk factors between the different stroke etiological subtypes. Lp(a) is independently associated with LAA and CE; however, it has no independent correlation with SAO.
4.Immune responses to recombinant hepatitis B virus vaccine in human immunodeficiency virus-1-infected patients with different CD4+ T-lymphocyte
Lin CAI ; Ruifeng ZHOU ; Yingchun ZHU ; Dongmei YAN ; Yan WANG ; Shenghua HE
Chinese Journal of Infectious Diseases 2014;32(3):139-143
Objective To compare the difference of immune responses to hepatitis B virus (HBV) vaccine in human immunodeficiency virus (HIV)-1-infected patients with different CD4+ T-lymphocyte counts.Methods HIV-1-infected patients who visited clinic at the Public Health Clinical Center of Chengdu were enrolled and divided into two groups according to CD4+ T-lymphocytes counts.CD4+ T-lymphocytes <200/μL,which were and ≥200/μL.All patients were injected intramuscularly with 20 μg of HBV vaccine at month 0,1 and 6,respectively.Vaccination responses were measured at1 and 7 months after first dose.The serum anti-hepatitis B sarface antigen titers of ≥ 10 mIU/mL were considered positive.The serum anti-HBs positive rates was compared by Chi-square test and anti-HBs titers was compared by Mann-Whitney test.Results Ninety-five HIV-1-infected patients were finally enrolled into the analysis,with 55 patients in group of CD4+ T-lymphocytes <200/μL and 40 patients in group of CD4+ T-lymphocytes ≥200/μL.The serum anti-HBs positivity rates at 1 and 7 months after first dose in group of CD4+ T-lymphocytes <200/μL were 40.0% (22/55) and 50.9% (28/55),respectively,which were 47.5% (19/40) and 75.0% (30/40) in group of CD4+ Tlymphocytes ≥200/μL.There was no significant difference of serum anti-HBs positivity rates between two groups after 1 month (x2 =5.652,P=0.017).But a significant higher positivity rate was found in group of CD4+ T-lymphocytes ≥200/μL than in group ofCD4+ Tlymphocytes<200/μL after 7 months (x2=0.531,P=0.466).The median titers of anti-HBs at 7 months were 10.44 mIU/mL in grouP of CD4+ T-lymphocytes < 200/μL and 222.73 mIU/ml in group of CD4+ T-lymphocytes ≥200/μL.There was significant difference of anti-HBs titers between two groups (Z=-3.600,P<0.01).No patients experienced side effects to HBV vaccine.Logistic regression model analysis indicated that only CD4+ Tlymphocytes was related to anti-HBs positivity after HBV vaccination.Conclusions HIV-1-infected patients with CD4+ T-lymphocytes ≥200/μL have higher serum anti-HBs positivity rates and higher anti HBs titers than patients with CD4+ T-lymphocytes <200/μL.However,patients with CD4+ T lymphocytes <200/μL also have relatively low immune response.Thus,HBV vaccination should be conducted in all these patients at high risk.
5.Simultaneous Determination of 6 Residual Organic Solvents in Aprepitant Raw Material by Headspace Capillary GC
Ruifeng ZHENG ; Chen YANG ; Fengying REN ; Hongqian JIA ; Lin RAN ; Qihui QIN ; Xiaojun GOU ; Ju FENG
China Pharmacy 2017;28(24):3426-3429
OBJECTIVE:To establish a method for simultaneous determination of 6 residual organic solvents in aprepitant raw material as methanol,ethanol,acetone,isopropyl alcohol,methyl tert-butyl ether and tetrahydrofuran.METHODS:Headspace capillary gas chromatography was adopted.The determination was performed on DB-624 capillary column using temperature programming.The temperature of injector port was 180 ℃,and flame ionization detector was used with temperature of 260 ℃.Nitrogen was used as carrier gas with flow rate 3.0 mL/min.The spilt ratio was 5 ∶ 1,and head-space injection volume was 1.0 mL.The head-space equilibrium temperature was set at 80 ℃,and equilibrium time was 40 min.RESULTS:The linear ranges of methanol,ethanol,acetone,isopropyl alcohol,methyl tert-butyl ether,tetrahydrofuran were 6.052-605.232 μ g/mL (r=0.999 9),9.987-998.718 μg/mL(r=0.999 9),9.998-999.768 μg/mL(r=0.999 8),9.986-998.634 μg/mL(r=0.999 9),9.991-999.090 μg/mL (r=0.999 7),1.461-146.133 μg/mL(r=0.999 5),respectively.The limits of quantitation were 1.782 1,2.079 0,0.749 8,1.777 8,0.223 1,0.607 0 μg/mL;the limits of detection were 0.594 0,0.693 0,0.249 9,0.592 6,0.074 4,0.202 3 μg/mL,respectively.RSD of precision test was lower than 2.0%.Only acetone and isopropyl alcohol were detected in stability test and reproducibility tests,RSD<2.0%.Their recoveries were 99.34-100.75% (RSD=0.52%,n=9),98.20%-100.24% (RSD=0.69%,n=9),98.07%-100.07% (RSD=0.84%,n=9),99.86%-101.32% (RSD=0.58%,n=9),97.87%-104.02% (RSD=2.13%,n=9),98.26 %-100.58 % (RSD =0.75 %,n =9),respectively.CONCLUSIONS:The established method is simple,accurate and reproducible,and can be used for simultaneous determination of 6 residual organic solvents in aprepitant raw material.
6.Porokeratosis: clinical analysis of seven cases
Hui CHEN ; Ruifeng SUN ; Chen ZHAO ; Yuying LIN ; Shi LIAN ; Wei ZHU
Chinese Journal of General Practitioners 2017;16(7):548-550
The clinical data of 7 patients with porokeratosis (PK) were analyzed retrospectively.In 7 PK patients, 4 cases were disseminated superficial actinic porokeratosis (DSAP),1 case was disseminated superficial porokeratosis (DSP),1 case was giant porokeratosis (GP) and 1 case was hypertrophic porokeratosis (HP).The characteristic cutaneous manifestations were annular well-circumscribed keratotic plaques with slightly atrophic center and elevated border.All patients shared a common histological hallmark, the cornoid lamella.Four cases of DSAP patients had family medical history, consistent with autosomal dominant inheritance.DSP, GP and HP patients denied family medical history.Diagnosis of PK should be based on clinical manifestations, family medical history and histopathological examination.
7.High Cell Density Culture of Lactobacillus plantarum Lp-2
Guangmin ZHANG ; Wei WANG ; Huifang BAO ; Xiaoling ZHANG ; Faqiang ZHAN ; Ruifeng LIN
China Biotechnology 2006;0(06):-
High cell density culture of Lactobacillus plantaru is a significant step of the preparation of lactic acid bacteria starter.Firstly,this experiment is designed to analyze the effects of different dissolved oxygen and pH on the batch fermentation of Lactobacillus plantarum.And then,on the base of the batch fermentation,fed-batch fermentation was studied,companied by chemic neutralization in the experiment to achieve the aim of high cell density cultivation.Finally,in the comparison of two kinds of feeding mode for fed-batch operation in 10L fermentor,it was demonstrated that two kinds of feeding mode made cell concentration increase differently,in which pH feedback feeding can control sucrose concentration within a certain lower range,therefore obtaining the maximum cell concentration.With the pH feedback feeding,the dry cell weight was up to 13.56g/L,which increased 90.05% over that of batch culture.
8.Circular RNAs in the pathogenesis of sepsis and their clinical implications: A narrative review.
Lin WEI ; Yongpeng YANG ; Weikai WANG ; Ruifeng XU
Annals of the Academy of Medicine, Singapore 2022;51(4):221-227
INTRODUCTION:
Sepsis is defined as a life-threatening complication that occurs when the body responds to an infection attacking the host. Sepsis rapidly progresses and patients deteriorate and develop septic shock, with multiple organ failure, if not promptly treated. Currently no effective therapy is available for sepsis; therefore, early diagnosis is crucial to decrease the high mortality rate. Genome-wide expression analyses of patients in critical conditions have confirmed that the expression levels of the majority of genes are changed, suggesting that the molecular basis of sepsis is at the gene level. This review aims to elucidate the role of circular (circ) RNAs in the pathogenesis of sepsis and sepsis-induced organ damage. In addition, the feasibility of using circRNAs as novel diagnostic biomarkers for sepsis is also discussed, as well as circRNA-based therapy.
METHOD:
This narrative review is based on a literature search using Medline database. Search terms used were "circular RNAs and sepsis", "circRNAs and sepsis", "non-coding RNAs and sepsis", "ncRNAs and sepsis", "circRNAs and septic pathogenesis", "circRNAs and septic model", "circRNAs and septic shock" and "circRNAs, biomarker, and sepsis".
RESULTS:
Numerous studies indicate that circRNAs might exert pivotal roles in regulating the immune system of the host against various pathogens, such as bacteria and viruses. Dysregulation of circRNA expression levels has been confirmed as an early event in sepsis and associated with the inflammatory response, immunosuppression and coagulation dysfunction. This impairment in regulation eventually leads to multiple organ dysfunctions, including of the kidneys, lungs and heart.
CONCLUSION
By investigating the regulation of circRNAs in sepsis, new molecular targets for the diagnosis and intervention of sepsis can be identified. Such an understanding will be important for the development of therapeutic drugs.
Biomarkers
;
Humans
;
RNA, Circular
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Shock, Septic/genetics*
9.MRI findings of the brain after gas explosion and its relationship with post-traumatic stress disorder
Ruifeng ZHAO ; Jilong JIN ; Huabing LI ; Shufeng LI ; Shuwen TIAN ; Haixue LI ; Yanhui CHEN ; Tianliang WANG ; Lin MA ; Zijing REN
Chinese Journal of Radiology 2008;42(12):1241-1245
Objective To investigate MR findings and dynamic changes of the brain after gas explosion,and to evaluate the relationship between MR findings and post-traumatic stress disorder (PTSD).Methods Forty-nine survivors of a gas explosion (group A) were examined with brain MRI within 1 to 3 days,and serial MR follow-up examinations were also performed.Forty miners not under the ground that day were assigned as group B,and 40 staff working on the ground as group C.The signal intensity values of hippocampus and globus pallidus on T2WI were measured in the three groups and F test was performed by using SPSS 13.0.The relationship between signal intensity values of hippocampus/globns pallidus and PTSD was explored,and the relationship between ADC values of hippocampus and PTSD was also investigated.Results In group A,slight low signal on T1WI and high signal on T2WI were detected on initial MRI in hippocampus (33 cases),globus pallidus (12 cases),cortex (10 cases),and midbrain (2 cases),respectively.On follow-up MRI at 2 months,lesions in hippocampus disappeared (25 cases) or remained slight high signal on T2WI (8 cases),lesions in globus pallidus disappeared (3 cases,5 sides) or showed shrinkage and encephalomalacia (9 cases),cortical lesions resulted in encephalomalacia in 2 cases and returned normal in the others,and lesions in the midbrain showed encephalomatacia.For comparison of T2 signal intensity values in hippocampus and globus pallidus,there was significant difference between group A and group B(P <0.01),and also between group A and group C(P <0.01),but no difference was detected between group B and group C (P>0.05).In group A,the T2 signal intensities of PTSD and non-PTSD were 455±37 and 462±53 in the left hippocarnpus,and 458±36 and 460±43 in the right hippoeampus on 1 to 3 days,and the T2 signal intensities of PTSD and non-PTSD were 438±29 and 424±37 in the left hippocampns,and 442±31 and 430±32 in the right hippocampus at 2 months.The T2 signal intensities of PTSD and non-PTSD were 361 ±35 and 366±63 in the left globus pallidus,and 363 ±41 and 375±62 in the right globus pallidus on 1 to 3 days,and the T2 signal intensities of PTSD and non-PTSD were 341±24 and 337±39 in the left globns pallidus,340±26 and 332±35 in the tight glohus pallidns at 2 months.There was no difference of T2 signal intensity values in hippocampus and globus pallidus between PTSD and non-PTSD( t=0.350,0.826,0.503,0.907,P>0.05).In group A,ADC values of PTSD and nun-PTSD were (8.1±1.1)×10-4 and(8.1 ±0.9)×10-4mm2/s in the left hippocampus,and (8.2±1.0)×10-4 and(8.2±0.8)×10-4mm2/s in the tight hippocampus on 1 to 3 days,ADC values were (8.8±0.7)×10-4 and (9.0±1.0)×10-4mm2/s in the left hippocampus,and (8.5±0.9)×10-4 and (9.3±1.1)×10-4mm2/s in the tight hippocampus at 2 months.ADC values in hippocampns showed no difference between PTSD and non-PTSD(t=0.016,0.081,P>0.05)on initial MRI,but showed significant difference between PTSD and non-PTSD in tight hippocampus (t=7.407,P < 0.05) on follow-up MRI at 2 months,while no difference in left hippocampus (t =0.333,P>0.05) was observed at 2 months.Conclusion Hippocampns and globus pallidus are the most vulnerable structures in gas explosion.The occurrence of PTSD may be related to the injury of fight hippocampus,but not related to the injury of globns pallidus.
10.AIDS patients with opportunistic infections in Sichuan province:clinical analysis of 1 465 cases
Shenghua HE ; Ruifeng ZHOU ; Dafeng LIU ; Lin CAI ; Yin WANG ; Xiaofei ZHOU ; Yi ZHANG ; Chunlan ZHANG ; Xiaoxia LIU ; Yao ZHANG
Chongqing Medicine 2014;(5):592-593,596
Objective To study clinical characters and prognosis of AIDS patients with opportunistic infections in Sichuan prov-ince .Methods We performed an retrospective analysis of 1 465 AIDS patients with opportunistic infections who were admitted into the transmitted disease hospital of Chengdu in recent 10 years .Results The overall mortality during hospitalization was 15 .49% . The leading cause of death was respiratory failure due to pneumonia (n=150 ,74 .62% ) or meningitis(n=47 ,23 .86% ) .Descending rank order of common opportunistic infections were respiratory tract ,the mouth swallows ,central nervous system ,gastrointestinal tract skin ,blood system .Descending rank order of common sites of infections were respiratory tract ,oropharynx ,central nervous system ,gastrointestinal tract skin ,reproductive tract .The multiple infections are common :more than 50% of patients suffering from two or more infections(50 .77% ) .Complexity of infection sites :42 .18% dual infection sites and 17 .20% three or more infec-tion sites .Conclusion The opportunistic infections disease spectrum of AIDS in Sichuan area has its own characteristics :multiple infections and multiple infections sites are both common ,death risks are high ,conditions of patients are severe generally ,and respir-atory failure is the main cause of death .