1.Advances in the Isolation Methods of Fungal Polyketide Synthase Genes
Shao-Hua YAN ; Liang GUO ; Yan-Ping LI ; Yang XU ;
China Biotechnology 2006;0(07):-
Fungal polyketide synthases are responsible for the biosynthesis of secondary metabolites such as pigments, mycotoxins, and they are very important in pharmacology , food science and agriculture. The recent advances in the methods for the isolation and manipulation of multiple classes of polyketide synthase genes from fungi were introduced. It is useful for discovery of novel fungal polyketide synthase gene clusters. These methods can also be useful for revealing the genetic potential of fungi to produce multiple types of bioactive polyketide.
3.Construction of cell model targeted on the damage by α-synuclein
Wenfen YAN ; Yang HENG ; Qianhang SHAO ; Naihong CHEN ; Yuhe YUAN
Chinese Pharmacological Bulletin 2015;(4):586-590
Aim To construct the cell model targeted on the damage by α-synuclein for screening anti-Parkinson’s Disease (PD)compounds.Methods The cDNA fragment of α-synucle-in gene was obtained by PCR methods and inserted into the re-combinant prokaryotic plasmid by molecular cloning technique. The recombinant plasmid was transformed into Escherichia coli, and subsequently induced to express α-synuclein protein.The recombinant α-synuclein was purified and identified by affinity chromatography,immunoblotting and mass spectrometry.The cells damage by α-synuclein was evaluated through cell viability measured by 3-(4,5-dimethyl-2-thiazolyl )-2,5-diphenyl-2-H-tetrazolium bromide.Results The obtained cDNA fragment ofα-synuclein in accordance with its theoretic molecular weight was cloned into pET30a plasmid and verified by sequencing.The re-combinant plasmid was transformed into bacteria E.Coli.BL21 (DE3)and induced to express α-synuclein by isopropyl β-D-1 -thiogalactopyranoside (IPTG).The expression condition was op-timized according to the culture temperature,the concentration of IPTG and the proliferation state of bacteria.The purified α-synu-clein was proved to be a 1 5.3 ku molecule weight protein,and could be immunoblotted with anti-α-synuclein antibody.The pu-rified α-synuclein could decrease the viability of PC1 2 cells and primary neurons significantly,and its effect was in a concentra-tion-dependent manner.Conclusion We have succeeded in constructing the cell model targeted on the damage by α-synucle-in.
4.Dynamic hip screw, proximal femoral nail antirotation and InterTan nail for intertrochanteric fractures
Jin SHAO ; Tieyi YANG ; Zhi WANG ; Yan ZHANG ; Shuyi LIU
Chinese Journal of Tissue Engineering Research 2015;(22):3543-3549
BACKGROUND: The extramedulary fixation system including dynamic hip screw (DHS) is commonly used in treatment of Intertrochanteric fracture. However, in patients with unstable intertrochanteric fracture, extramedulary fixation system often leads to the failure of fracture fixation. Intramedulary fixation system including both proximal femoral nail antirotation (PFNA) and InterTan nail has been widely used in the treatment of unstable intertrochanteric fractures. OBJECTIVE:To compare the therapeutic effects of extramedulary fixation system containing DHS, PFNA and InterTan nail in the treatment of intertrochanteric fracture. METHODS:Literatures were searched in Wanfang, PubMed, Embase, Medline, the Cochrane library to screen literatures published from January 1990 to November 2014. Relevant studies addressing extramedulary fixation system containing DHS, PFNA and InterTan nail were screened. RESULTS AND CONCLUSION: 346 articles were screened, and 13 of them were in accordance with the inclusion criteria. 1 271 patients with different types of intertrochanteric fracture were assessed in this study. Compared to DHS group, patients treated with PFNA and InterTan nail had shorter operation time and less blood loss. No significant difference in rehabilitation time and Harris score was detected among three kinds of fixation methods. Additionaly, PFNA and InterTan nail had a similar effect. These findings verify that compared with DHS, PFNA and InterTan nail can optimize the surgery, but cannot elevate postoperative outcomes.
5.Demonstration of nested case-control study design in mechanisms research of allergic reaction of tanreqing injection.
Peng-Yang XIE ; Yan-Ming XIE ; Lian-Xin WANG ; Yan-Peng CHANG ; Shao-Yong LIU
China Journal of Chinese Materia Medica 2014;39(18):3567-3570
The paper is focused on the clinical applications of Tanreqing injection after listing, detecting and analyzing the related blood indicators of patients with allergic reactions based on prospective, multi-center, large sample, registration-type clinical safety monitoring nested case-control study (NCCS) to explore the possible mechanisms of allergic reaction of Tanreqing injection, 3 006 patients cases used with Tanreqing injection were monitored, including 3 cases of adverse reactions and 2 cases of allergic reactions. Each patient of allergic reactions, according to the ratio of 1:4 matches four cases of not adverse reactions as a control group of patients, while 5 healthy and 5 cases of volunteers into the healthy group. We examined the correlation detection of cases of allergic reactions among groups such as T-IgE, IgA, IgG, C3, C4, IFN-g, IL-2, IL-4, IL-6, IL-10. Allergic reactions of Tanreqing injection may be mediated by IgE as type I based on existing research data. This results and conclusions will promote the justifiability and safety of clinical applications.
Adolescent
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Adult
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Case-Control Studies
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Drugs, Chinese Herbal
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administration & dosage
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adverse effects
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Female
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Humans
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Hypersensitivity
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diagnosis
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metabolism
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Interleukin-10
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metabolism
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Interleukin-2
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metabolism
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Interleukin-4
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metabolism
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Interleukin-6
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metabolism
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Male
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Prospective Studies
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Young Adult
6.Electrophysiological study of V535M hERG mutation of LQT2.
Chunyan, SHAO ; Yan, LU ; Mohan, LIU ; Qi, CHEN ; Yunfeng, LAN ; Yan, LIU ; Min, LIN ; Yang, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(6):741-8
This study examined the current changes of human ether-a-go-go-related gene (hERG) mutation derived from a LQT2 Chinese family with a highly penetrating phenotype. Mutation was identified and site-directed mutagenesis was performed to induce the mutation in wild-type (WT) hERG. WT hERG and mutated V535M were cloned and transiently expressed in HEK293 cells. At the 48th and 72nd h after transfection, membrane currents were recorded using whole cell patch-clamp procedures. An A>G transition at 1605 resulting in replacement of V535M was identified. Compared to WT, V535M mutation significantly decreased tail currents of hERG. At test potential of -40 mV after depolarizing at +50 mV, tail current densities were 83.35±7.06 pA/pF in WT and 50.38±7.74 pA/pF in V535M respectively (n=20, P<0.01). Gating kinetics of hERG revealed that V (1/2) of steady-state inactivation shifted to negative potential in the mutant (V (1/2,V535M): -61.81±1.7 mV vs. V (1/2, WT): -43.1±0.71 mV). The time constant of recovery from inactivation was markedly prolonged in the mutant compared to WT among test potentials. V535M hERG mutation demonstrated markedly decreased tail current densities, which suggests that V535M is a new loss-of-function mutation of hERG channel responsible for LQT2.
7.Biosynthesis and assay of crude T-2 toxin
Yan-hong, CAO ; Shao-ping, WANG ; Yang, HUI ; Xiao-yan, FU ; Wei, DONG ; Ning, LIU
Chinese Journal of Endemiology 2008;27(5):552-554
Objective To isolate Fusarium species from Kashin-Beck disease(KBD)area and biosynthesize cnlde T-2 toxin.Methods T-2 toxin.producing Fusarium was isolated from corns produced in KBD area and purifted.The purifted funsi were identified according to the traits of colony,appearance of thallus and characters of conidium and then weIe cultivated in sterile Corn culture media.After extraction with organic solvent and purification by silica gel chromatography column,the quality and quantity of the toxin in the extracts were estimated by thin,Layer chromatography and high-performance liquid chromatography.Results The toxin-producing strain was Fusarium tricinctum. The com cuIture media inoculated with this strain produced about 250 mg of crude T-2 toxin per kg. Conclusions This experiment has indirectly further confirmed pollution of T-2 toxin-producing Fmarium existed in
8.Vitamin D status and its associated risk factors among healthy adults in Gansu province
Wenjuan MA ; Jing LIU ; Lianhua WEI ; Yan YANG ; Ruifei YANG ; Feifei SHAO ; Yu WANG ; Limin TIAN
Chinese Journal of Clinical Nutrition 2017;25(4):240-245
Objective To investigate the prevalence of vitamin D deficiency and its associated metabolic risk factors in the healthy adults in Gansu Province, a western province of China.Methods Through stratified, multistage probability population sampling, 11 300 adults (4 740 men and 6 417 women aged 18-79 years) who had been living in their current residence for at least 5 years were arbitrarily chosen from seven representative geographical cities in Gansu Province from June 2014 to December 2015.The data obtained from a questionnaire-based survey, anthropometric and biochemical assessments, and areal bone mineral density measurement were collected and analyzed.Results The overall mean serum 25(OH)D was (16.07±9.40) μg/L.The prevalence of severe deficiency (<10 μg/L), deficiency (≥10 μg/L and <20 μg/L), insufficiency (≥20 μg/Land <30 μg/L), and sufficiency (≥30 μg/L) among these adults were 17.3%, 64.6%, 11.8%, and 6.3%, respectively.The significant predictors of vitamin D deficiency included younger age and female sex(P<0.05), whereas sun exposure, physical activity, and calcium supplementation were protective (P<0.05).Serum 25(OH)D was inversely associated with parathyroid hormone (r=-0.279, P<0.001) and positively associated with serum calcium (r=0.239, P<0.001), serum phosphorus (r=0.090, P=0.018), low-density lipoprotein cholesterol(r=0.100, P=0.008), and body mass index (r=0.093, P=0.014).The 25(OH)D concentration showed no significant correlation with bone mineral density (P>0.05).Conclusions Vitamin D deficiency is highly prevalent among adult populations in Gansu, northwestern of China, and is largely attributed to younger age and female sex.The cumulative effects of factors including sun exposure, physical activity, and calcium supplementation play an influential role in maintaining vitamin D levels.
9.Clinical observation of HBV reactivation and prevention in renal allograft recipients with inactive HBsAg carriers
Tianbiao LAN ; Xingfeng REN ; Yan CHEN ; Jun PENG ; Cheng YANG ; Ziwen SHAO
Chinese Journal of Organ Transplantation 2012;(10):602-605
Objective To investigate the HBV reactivation status and clinic outcomes in the renal allograft recipients with inactive HBsAg carriers,and explore the preventive measures.Methods A retrospective analysis of clinical manifestation was processed in 88 cases of inactive HBsAg carriers before and after renal transplantation.Preoperative liver function in all cases was normal and serum HBsAg positive,HBV DNA<106 copies/L.Tacrolimus (or cyclosporine A) + mycophenolate mofetil (MMF) + prednisone were given in prevention of rejection after transplantation.In 88 cases,56 cases were given nucleoside analogues (acid) for prophylactic antiviral therapy,in which 31 cases were given lamivudine (LAM) (LAM group),25 cases were given entecavir (ETV) (ETV group) ; The rest 32 cases were not given prophylactic antiviral therapy,only receiving routine liver-protecting therapy (inosine,glucurolactone) (control group).Incidence of HBV re-activation,liver function,response to treatment and the pathological changes of hepatic tissue were observed.Results During the follow-up period,the incidence of HBV reactivation in LAM group and ETV group was 45.2% and 28.0% respectively,significantly lower than in control group (84.4%,P< 0.05).In prophylactic treatment groups,HBV reactivation occurred later,liver function damage was milder,and HBV DNA load peak was lower (P<0.05).In LAM group,HBV reactivation occurred in 14 cases,including 10 cases occurred during administration of LAM,and ETV treatment was given for about 2 months,serum HBV DNA levels in 7 cases were under detection line;in the rest 4 cases,HBV reactivation occurred in patients with treatment less than 1 year and noncompliance,who withdrew medicine blindly.After the original scheme of antiviral therapy was done,serum HBV DNA levels in 3 cases were under detection line,and the effect was not obvious in one case.In control group,HBV reactivation occurred in 27 cases.Fourteen cases therefore accepted nucleoside (acid) analogs antiviral therapy,and HBV DNA levels in 10 cases were under detection line.Histological examination revealed the liver with fibrotic cholestatic hepatitis changes in 9 patients,including 8 cases in control group,and 1 case in LAM group due to blind withdrawal of medicine.Conclus(i)on LAM and ETV prophylactic use may decrease the HBV reactivation rate in inactive HBsAg carriers after renal transplantation,reduce the severity of liver damage and the occurrence of fibrotic cholestatic hepatitis.
10.An analysis of intervention outcome in non-ST segment elevation acute coronary syndrome in elderly patients
Jun ZHANG ; Shubin QIAO ; Jun ZHU ; Jue CHEN ; Weixian YANG ; Yan LIANG ; Chunli SHAO
Chinese Journal of Internal Medicine 2011;50(5):378-382
Objective To investigate the effect and safety of early intervention and delayed intervention therapy on elderly patients and younger patients with non-ST segment elevation acute coronary syndrome. Methods The patients with non-ST segment elevation acute coronary syndrome were randomly divided into early intervention group (coronary angiography taken within 24 hours after grouping) and delayed intervention group ( coronary angiography taken after 36 hours after grouping). The primary endpoint was a composite endpoint of death, myocardial infarction and stroke during 180 days follow-up. Results A total of 815 patients were enrolled, including 198 elderly patients aged 75 years and above, and 617 younger patients aged below 75 years. The elderly patients had a greater incidence of the primary endpoint than that of younger patients ( P = 0. 00). The primary endpoint of early intervention group were obviously lower than that of delayed intervention group of younger patients ( P = 0. 01 ). There was no significant difference in primary endpoint incidence of early intervention group and delayed intervention group of the elderly patients (P =0. 39). Conclusions The elderly patients with non-ST segment elevation acute coronary syndrome who underwent intervention had greater incidence of death and myocardial infarction. Early intervention reduced the rate of myocardial infarction for the younger patients. There was no significant difference in primary endpoint incidence between early intervention and delayed intervention among elderly patients.