1.Effects of hyperacetylation of H3K9ac mediated by histone acetylases on the overexpression of MEF2C induced by alcohol in the hearts of fetal mice during pregnancy
Chang PENG ; Xiaomei LUO ; Hongbo XU ; Xiangli LI ; Jie TIAN
Chinese Journal of Applied Clinical Pediatrics 2016;(1):37-41
Objective To investigate the effects of H3K9ac hyperacetylation mediated by histone acetylases on the overexpression of MEF2C in the hearts of fetal mice exposed to alcohol during pregnancy,and provide new interven-tion targets for prevention and treatment of cardiac dysplasia caused by alcohol exposure.Methods C57BL/6 mice were divided into 5 groups randomly,blank control group,dimethylsulfoxide (DMSO)group,alcohol group,alcohol +anacardic acid group and anacardic acid group,and then the hearts of fetal mice were collected to be analyzed.Chroma-tin immunoprecipitation and Western blot were used in assaying the binding of histone acetylases and the level of H3K9ac to the promoter of MEF2C in the hearts of fetal mice.The mRNA expression of MEF2C was tested by adopting real -time PCR.Results The level of H3K9ac in the promoter of MEF2C in the hearts of fetal mice exposed to alcohol was higher than that in the hearts of fetal mice exposed to saline (1 .30 ±0.1 9 vs 0.45 ±0.01 ),there was statistically significant difference (P <0.05),while the binding of E1 A -binding protein (p300),p300 /cyclic adenosine mono-phosphate response element binding protein -associated factor (PCAF)and steroid receptor coactivator -1 (SRC1 )to the promoter of MEF2C were abnormally elevated in the hearts of fetal mice treated by alcohol (1 .68 ±0.08 vs 0.82 ± 0.08,1 .08 ±0.05 vs 0.42 ±0.02,1 .1 8 ±0.05 vs 0.39 ±0.08),and there were statistically significant differences (all P <0.05).The expression of MEF2C mRNA in alcohol group was higher than that in blank control group (1 .36 ± 0.1 2 vs 0.29 ±0.03),there was statistically significant difference(P <0.05).However,a pan -acetylases inhibitor, anacardic acid,could decrease significantly the binding of p300 and PCAF to the promoter of MEF2C (1 .52 ±0.05 vs 0.63 ±0.09,1 .1 3 ±0.04 vs 0.45 ±0.04),and correct abnormal hyperacetylation of H3K9ac induced by alcohol (1 .58 ±0.08 vs 0.67 ±0.05),and down -regulate the over -expression of MEF2C in the hearts of fetal mice exposed to alcohol (1 .36 ±0.1 2 vs 0.41 ±0.05 ),and there were statistically significant differences (all P <0.05 ). Conclusions Hyperacetylation of H3K9ac mediated by p300 and PCAF may be a key regulatory factor in the over -expression of cardiac nuclear transcription factor MEF2C in the hearts of fetal mice exposed to alcohol during pregnan-cy.Anacardic acid can significantly attenuate the level of H3K9ac through inhibiting the binding of p300 and PCAF to the promoter of MEF2C,and down -regulate the over -expression of cardiac nuclear transcription factor MEF2C in the hearts of fetal mice.
2.Total hip arthroplasty for post-traumatic hip fusion ankylosis secondary to operation in acetabular combined with femoral head fracture
Ping ZHEN ; Xusheng LI ; Qi TIAN ; Shenhu ZHOU ; Yanfeng CHANG
Chinese Journal of Orthopaedics 2016;36(19):1213-1221
Objective To investigate the technique and clinical results of total hip arthroplasty in treating post-traumatic hip fusion ankylosis secondary to operation in acetabular combined with femoral head fracture.Methods From October 2009 to January 2015,7 patients (7 hips) with post-traumatic hip fusion ankylosis underwent total hip arthroplasty.There were 6 males and 1 female with an average age of 38 years (range,25-51 years).There were 4 hips on the left side,3 hips on the right side.Open reduction and internal fixation were performed on all patients.One patient had a postoperative deep infection and 6 patients underwent implant removal.The hip bony fusion ankylosis was developed secondary to post-traumatic arthritis after the surgery of ipsilateral acetabular combined with femoral head facture.The interval between internal fixation and total hip arthroplasty was 37.6 months on average (range,21-67 months).The hip bony fusion ankylosis was relieved by wedge shape osteotomy at femoral neck level.The cemenfless prostheses were implanted in suitable places based on the accuracy position of acetabular center.One infection case was treated via surgical debridement,removal of all the screws,and antibiotic-loaded cement spacer implantation in the first stage.The cementless prostheses were implanted in the second stage.Results The average operation time and blood loss was 108 min (90-150 min) and 320 ml (280-450 ml) respectively.The internal fixation devices were explanted completely,including plate and screw in 5 cases,only screw in 1 case,intramedullary nail and screw in 1 case.The mean fallow-up period was 32.5 months (range,15-48 months).The average Harris hip score improved from 48.8±6.5 points preoperatively to 92.6±5.1 points post-operatively (t=22.82,P=0.001).Osseointegration was developed in all of the acetabular and femoral components at 3 months post-operatively.Radiograph analysis showed satisfied position of acetabular cup and no evidence of implant migration or center of rotation change.Stem subsidence (<1.0 mm) occurred in 2 cases and heterotopic ossification in 2 cases (Brooker type Ⅰ and type Ⅱ in 1 case,respectively) at one year postoperatively.No complication occurred,such as damages of vessels and nerve,dislocation,component loosening or infection.Conclusion The wedge shape osteotomy of femoral neck is a safe and reliable method in treating hip fusion ankylosis.Total hip arthroplasty can be achieved by reconstruction of acetabular center,suitable components implanted in anatomical position and ideal reconstruction of soft tissue around hip.
3.Inhibitory effects of extracellular cholesterol and lipopolysaccharide on cellular cholesterol efflus
Jie LIU ; Yunmei ZHENG ; Zhihui TIAN ; Guangming CHANG ; Haidong LI
Chinese Journal of Tissue Engineering Research 2016;20(7):1000-1005
BACKGROUND:Cholesterol is closely linked to the occurrence and progression of atherosclerosis. Current approaches to study celular cholesterol dynamics have their own limitations.
OBJECTIVE: To measure the cholesterol efflux rate of RAW 264.7 mouse macrophages by BODIPY-Cholesterol labeling and to explore the effects of extracelular cholesterol and lipopolysaccharide on the cholesterol efflux rate.
METHODS:RAW 264.7 cels were cultured in vitro with DMEM containing 10% fetal bovine serum, and labeled with BODIPY-Cholesterol for 1, 2, 4, 8 hours. Then, the cels were rinsed with serum-free DMEM and inoculated for 6, 12, 24, 48, 96 hours to optimize the labeling time and incubation time. We measured and compared the cholesterol efflux rates after cultured cels were treated with cholesterol, lipopolysaccharide, human sera with high cholesterol or human sera with normal cholesterol.
RESULTS AND CONCLUSION: The best labeling time for BODIPY-Cholesterol was 2-8 hours. Cholesterol efflux rates were gradualy decreased after the cels that were labeled for 2 hours were incubated with increasing concentrations of cholesterol (0.1, 0.5, 2.5 mmol/L,P< 0.01). Treating cels with lipopolysaccharide also decreased the cholesterol efflux rate (P < 0.05). Furthermore, the cholesterol efflux rate was decreased after cels were treated with human sera with high cholesterol (P< 0.05). These findings indicate that BODIPY-Cholesterol can be used to measure celular cholesterol efflux rate and to study the effects of extracelular cholesterol and lipopolysaccharide on the cholesterol efflux rate.
4.Construction of human PPAR?_1 LBD fusion protein expression vector and optimization of inducing conditions
Mi TIAN ; Changqing LI ; Wei CHANG ; Lingyun SHI ; Qixin ZHOU
Chinese Pharmacological Bulletin 2003;0(09):-
Aim To obtain high pure hPPAR?1LBD fusion protein.Methods A cDNA encoding ligand binding domain(LBD)of PPAR?1 was amplified by RT-PCR from human fatty tissue and the product was inserted into the downstream of the malE gene in the vector pMAL-p2X,which encoded maltose-binding protein(MBP).The recombinant plasmid containing MBP-PPAR?1 gene was transformed into E.coli.TB1 and the expression conditions of the recombinant strain were optimized.Results The DNA strap of MW(909 bp) was presented after re-combinant plasmid was digested by Hind Ⅲ and BamH Ⅰ.The high efficient expression of MBP-PPAR?1 fusion protein in TB1 cells was observed with 38.54% product of the total cytoplasm proteins when 0.4 mmol?L-1 IPTG and 6 h incubation were taken at 30℃.Conclusion The recombinant vector was successfully constructed.It could high efficiently express hPPAR?1LBD fusion protein in TB1 cells and obtain the hPPAR?1LBD fusion protein with high bioactivity.
5.Epidemiological investigation of 235 patients with extra-pulmonary tuberculosis wounds.
Na CHANG ; Chiyu JIA ; Zhen LIU ; Yajie ZHANG ; Wenting LI ; Tian TIAN
Chinese Journal of Burns 2015;31(2):122-124
OBJECTIVETo investigate the epidemiological characteristics and patterns of extra-pulmonary tuberculosis wounds in order to provide reliable data for further clinical research.
METHODSRecords of patients with extra-pulmonary tuberculosis wounds hospitalized from January 2010 to December 2012 were retrospectively analyzed, including gender, age, nationality, family background, Bacille Calmette-Guerin (BCG) vaccination, primary lesion, and history of injury.
RESULTSTuberculosis wounds were found in 235 patients among 5 863 patients with extra-pulmonary tuberculosis, accounting for 4.0%. Among the patients with tuberculosis wounds, there were 139 male and 96 female, and the ratio of male to female was 1.4: 1.0. The age of patients ranged from 1 to 87 (37 +/- 18) years old, and the highest incidence occurred in patients older than 15 and younger than or equal to 30 years old (100 cases, accounting for 42.6%). Most patients with tuberculosis wounds were Han, and only 11 patients were minorities, accounting for 4.7%. Tuberculosis wounds were more prevalent in rural areas (163 cases, accounting for 69.4%), with a smaller number in urban areas (72 cases, accounting for 30.6%). The BCG vaccination rate was 13.6%. The main primary lesions were lymph node infection (112 cases, accounting for 47.7%), among which involvement of cervical lymph nodes accounted for the highest ratio ( 99 cases, accounting for 88.4%). Twenty-one patients had the traffic accident etc. injury history recently, among which 19 were male and 2 were female.
CONCLUSIONSTuberculosis wound, with certain incidence, was more frequently found among young adults from rural areas. The BCG vaccination rate was low among the patients and the main primary lesion was tuberculosis of cervical lymph nodes.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; ethnology ; statistics & numerical data ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Incidence ; Infant ; Lymph Nodes ; microbiology ; Male ; Middle Aged ; Population Surveillance ; Prevalence ; Retrospective Studies ; Rural Population ; Tuberculosis ; epidemiology ; microbiology ; Tuberculosis, Pulmonary ; epidemiology ; Urban Population ; Wounds and Injuries ; complications ; epidemiology ; Young Adult
6.The value of the timed “up and go” test at the evaluation of functional mobility in stroke patients
Chang-shui WENG ; Zhe TIAN ; TIAN LI ; Suqing BI ; Jun XU ; Zengzhi YU ; Chunnuan HUO ; Liping GAO ; Jun WANG
Chinese Journal of Rehabilitation Theory and Practice 2004;10(12):733-735
ObjectiveTo evaluate the effect of the timed “up and go” test (TUGT) on measuring functional mobility of stroke patients.MethodsNinety hemiparetic stroke patients participated in this study. The balance, gait speed and disability of patients were measured by Berg balance scale (BBS), maximal gait speed and functional independence measure (FIM) to find out the critical value of TUGT.ResultsA good relationship existed among TUGT and the BBS,gait speed and FIM (r=-0.926—-0.674,P<0.001).The percentage of independent walking of stroke patients whose TUGT scores <10s or>20s were 100% and 8.3%. The optimal cut off values of TUGT to predict the independent walking of patients were 15.2s, and in stroke group sensitivity and specificity of TUGT were 89.4% and 79.1%.Conclusion TUGT is a reliable instrument with adequate concurrent validity to measure the functional mobility of stroke patients.
7.Protective Roles of Melatonin against Acrylamide-induced Testicular Toxicity in Rats
Yuxin MA ; Sumin TIAN ; Jing LIU ; Min ZHENG ; Chang ZHOU ; Jie ZHANG ; Li LUO ; Guoying LI
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(4):517-525
[Objective] The present study aimed to determine the protective role of Melatonin (MT) against Acrylamide (AA)-induced testicular toxicity and the potential molecular mechanism.[Methods] The animals were randomly divided into three groups,the control group (n =12),the AA group (n =12) and the AA+MT group (n =12).The rats in the AA and AA + MT group were gavaged with AA at a dose of 15 mg/(kg · day) for 4 consecutive weeks.After 3 weeks of AA treatment,MT was intraperitoneally injected 30 minutes before AA treatment at 10 mg/(kg· day) for 1 week in the AA + MT rats.Subsequently,the mitochondrial membrane potential measurement,TUNEL assay,Western blot and electron microscopic techniques were applied in the present study.[Results] The results showed that AA could decrease the testis mitochondrial membrane potential (P < 0.05) which could be recovered by MT (P < 0.05).Moreover,MT induced down-regulation of Bax expression and up-regulation of Bcl-2 expression in the testis,compared with AA rats.The amelioration of testicular apoptosis was further confirmed by the TUNEL labeling.Western blot results suggested that the decreased ratios of Bcl-2/Bax and Bcl-xL/Bak in the AA group (both P < 0.05) could be recovered by MT treatment (both P < 0.05).The levels of Cyt-c,Casp-3,p53 and NF-κB in AA group were markedly elevated compared with the control (all P < 0.05),and reduced in MT treatment group (all P < 0.05).MT could relieve abnormal mitochondrial structures in the seminiferous tubule in the electron microscopic level.[Conclusion] MT may exert productive effect through its anti-apoptotic properties associated with mitochondria.
8.Colonization characteristics of endophytic bacteria NJ13 in Panax ginseng and its biocontrol efficiency against Alternaria leaf spot of ginseng.
Chang-Qing CHEN ; Tong LI ; Xin-Lian LI ; Yun JIANG ; Lei TIAN ; Peng XU
China Journal of Chinese Materia Medica 2014;39(10):1782-1787
To reveal the colonization characteristics in host of endophytic biocontrol bacteria NJ13 isolated from Panax ginseng, this study obtained the marked strain NJ13-R which was double antibiotic resistant to rifampicin and streptomycin through enhancing the method of inducing antibiotic. The colonization characteristics in ginseng and its biocontrol efficiency against Alternaria spot of ginseng in the field were studied. The results showed that the strain could colonize in root, stem and leaf of ginseng and the colonization amount was positive correlated with inoculation concentration. Meanwhile, the strain could infect and then transfer in different tissues of ginseng The colonization amount of strain in roots and leaves of ginseng increased first and then decreased. However, the tendency of colonization amount of strain in stems was ascend at first and then descend slowly, and was more than that in roots and leaves along with time, which had a preference to specific tissue of its host. In field experiment, the endophytic bacteria NJ13 was proved to be effective in controlling Alternaria leaf spot of ginseng. The biocontrol efficiency of fermentation broth at the concentration of 0.76 x 10(8) cfu x mL(-1) reached 75.62%, which was close to the controlling level (73.06%) of 0.67 mg x L(-1) 50% cyprodinil WG.
Alternaria
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physiology
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Antibiosis
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Bacillus
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growth & development
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isolation & purification
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physiology
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Endophytes
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growth & development
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isolation & purification
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physiology
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Panax
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growth & development
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microbiology
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Plant Diseases
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microbiology
9.Clinical and pathologic characteristics of 4 cases of Wegener's granulomatosis associated with specific pathogenic infections.
Ying-shi PIAO ; Cheng TIAN ; Xue LI ; Chang-li YUE ; Hong-gang LIU
Chinese Journal of Pathology 2012;41(2):123-124
Adolescent
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Adult
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Aspergillosis
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complications
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microbiology
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Aspergillus
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isolation & purification
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Candida albicans
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isolation & purification
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Candidiasis
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complications
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microbiology
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Female
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Follow-Up Studies
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Granulomatosis with Polyangiitis
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complications
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microbiology
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pathology
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Humans
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Male
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Middle Aged
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Mucor
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isolation & purification
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Mucormycosis
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complications
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microbiology
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Nocardia
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isolation & purification
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Nocardia Infections
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complications
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microbiology
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Retrospective Studies
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Young Adult
10.A preliminary study on relationship between single nucleotide polymorphism ofmiR-196a2 and risk of leukemia
Wenjuan ZHANG ; Zhe TIAN ; Youjie LI ; Wenjing SHANG ; Ning XIE ; Jing CHANG ; Jing XIAO ; Xuelin BAO
China Oncology 2016;26(10):826-830
Background and purpose:miR-196a2 functions as an oncogene during tumor initiation and pro-gression. The up-regulation promotes tumor cell proliferation, invasion and metastasis. Therefore, it is promising to be an important tumor biomarker. The aim of this study was to investigate whether rs11614913, a gene polymorphic site ofmiR-196a2, is associated with the risk of leukemia.Methods:A case-control analysis was employed. Bone marrow or periph-eral blood was collected from 210 leukemia patients diagnosed from Jan. 2009 to Jul. 2015 in Yantaishan Hospital (case group) as well as 250 healthy people who were physically examined during the same period (control group). Polymerase chain reaction-restriction fragment length polymorphism (PCR-PFLP) was used to detect the genotype of rs11614913. Application test was used to compare the difference in the frequency of each genotype between case group and control group. The odds ratio (OR) of SNP allelic genes was calculated using logistic regression analysis and 95%CI represented the risk of leukemia for each genotype.Results:The distribution differences in the frequency of T/T, C/C, C/T genotype of miR-196a2 rs11614913 between case group and control group were statistically significant (P<0.05). The risk of leukemia for individuals who carried mutant homozygous C/C was 2.661-fold higher than those carried wild-type homozygous T/T, and the difference was statistically significant (P<0.05).Conclusion:ThemiR-196a2 gene polymorphic site rs11614913 was associated with the risk of leukemia. Mutant homozygous C/C or C allelic gene carrying was probably a risk factor for leukemia.