1.Investigation on gene mutation from hereditary protein S deficiency pedigree
Fang YANG ; Guanjun WANG ; Lihua KANG ; Xuefeng WANG ; Qiulan DING ; Hongli WANG
Chinese Journal of Laboratory Medicine 2010;33(6):517-521
Objective To identify the clinical phenotypic diagnosis and gene mutation detection of two kindreds with PS deficiency. MethodsPS: A was measured by chromogenic substrate method;TPS:Ag, FPS: Ag levels were measured by ELISA method; PS gene(PROS1 gene)was detected by amplifying 15 exons and flanking intron sequences from the propositus with PCR method. PCR products were purified and directly sequenced. Results For propositus 1,PS: A was 48.6% ,TPS: Ag was 136 mg/L, FPS : Ag was 41 mg/L, PROSI gene exon 2 was in c. Heterozygous base substitutions was detected in C121T locus, which led to Arg-1Cys (R-1C) heterozygous roissense mutation encoded in PS proteins. For propositus 2, PS: A was 29.2%, TPS: Ag was 83 mg/L, FPS: Ag was 26 mg/L, PROSI gene exon 14 was in c. Heterozygous base substitutions was identified in CI687T locus, in which Gln.522Stop heterozygous nonsense mutation was encoded in PS proteins. Conclusions c. C121T is a novel mutation locus detected in PROS1 gene. This heterozygous mutation could lead to type Ⅱ PS hereditary deficiency, while c. C1687T heterozygous mutation could bring about type Ⅰ PS hereditary deficiency.
2.An antimicrobial resistance surveillance of gram-positive cocci isolated from 12 teaching hospitals in China in 2009
Hongli SUN ; Hui WANG ; Minjun CHEN ; Yingmei LIU ; Zhidong HU ; Kang LIAO ; Yunzhuo CHU ; Jine LEI ; Bing ZHANG ; Yunsong YU ; Bijie HU ; Ziyong SUN ; Zheng ZHANG ; Qiyong HE
Chinese Journal of Internal Medicine 2010;49(9):735-740
Objective To investigate antimicrobial resistance among gram-positive cocci in China in 2009. Methods From June to December 2009, 1169 consecutive and non-repetitive gram-positive cocci were collected from 12 teaching hospitals at 9 cities. The minimal inhibitory concentration (MIC) of antibacterial agents was determined by agar dilution method. Results The prevalences of methicillinresistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococci (MRCoNS) were 45.3% (211/466) and 89. 5% (214/239), respectively. The isolation rate of MRSA was 33. 3%-68. 1% from different samples. All Staphylococci isolates were susceptible to vacomycin, teicoplanin and linezolid. Five point five percent (7/128) E. faecium strains were resistant to vacomycin. All E.faecalis strains were susceptible to vacomycin. About 99. 1% (108/109) of E. faecalis and E. faecium were susceptible to linezoild. The prevalence of penicillin-intermediate Streptococcus pneumoniae (PISP) was 21.6% (48/222). Only 1 (0. 5%, 1/222) Streptococcus pneumoniae strain was resistant to penicillin.Teicoplanin, vancomycin, linezolid and tigecycline were the most active agents against Streptococcus pneumoniae (susceptible rate 100% ). Conclusions The high prevalence of methicillin-resistance is among Staphylococcus strains. Different samples show a different MRSA prevalence. Teicoplanin, vancomycin and linezolid show very high activity to Staphylococci,E. faecalis, E. faecium and Streptococcus pneumoniae.
3.Antimicrobial resistance surveillance of gram-positive cocci isolated from 12 teaching hospitals in China in 2008
Hongli SUN ; Hui WANG ; Minjun CHEN ; Ziyong SUN ; Yunsong YU ; Bijie HU ; Yunzhuo CHU ; Kang LIAO ; Jine LEI ; Bing ZNANG ; Bin CAO ; Qiyong HE ; Zheng ZHANG ; Zhidong HU
Chinese Journal of Laboratory Medicine 2010;33(3):224-230
Objective To investigate antimicrobial resistance among gram-positive cocci in China in 2008.Methods From June 2008 to December 2008,1171 consecutive and non-repetitive gram-positive cocci were collected from 12 teaching hospitals.The MICs of antibacterial agents was determined by agar dilution method.Results The prevalence of MRSA and methicillin-resistant coagulase-negative Staphylococci(MRSCoN) was 49.9%(232/465) and 74.0%(179/242),respectively.The MRSA prevalence ranged from 33.3% to 65% in different regions.About 71.1%(108/152) of Staphylococcus aureus from respiratory tract specimens,48.3%(28/58) of Staphylococcus aureus from blood samples,and 36%(68/189) of Staphylococcus aureus from the pus,wound and sterile body fluid samples were resistant to methicillin.The susceptible rates of MRSA to trimethoprim/sulfamethoxazole(SXT) and chloramphenicol were 81.5%(183/232) and 89.7%(208/232).Susceptibility to gentamicin, erythromycin, clindamycin, tetracyclines,rifampicin,and quinolones were from 3.9% to 35.0%.All Staphylococci isolates were susceptible to vancomycin,teicoplanin,and linezolid.Three vacomycin-resistant Enterococcus faecium strains were found in this study.About 96.2%(101/105) of Enterococcus faecalis and 97%(130/134) of Enterococcus faecium were susceptible to linezoild.Fifty-one out of 105 of Enterococcus faecalis(48.6%)and 101 out of 134 Enterococcus faecium(75.4%)were resistant to high concentration gentaroicin.The susceptibility of Enterococcus faecalis to all the antibiotics except for chloramphenicol and tetracycline was higher than that of Enterococcus faecium.Enterococcus faecium isolates showed a high resistant prevalence to most of antibiotics except glycopeptides and linezolid.The prevalence of PISP among 225 isolates was was 36.6%(15/41),and the prevalence of PNSSP from the other patients ranged from 15.4% to 26.6%.The susceptible rates of PSSP to cefprozil,cefuroxime and cefaclor were 67.5%(114/169),66.3%(112/169) and 61.5%(104/169),respectively.All the PISP isolates were resistant to the above three antibiotics.Teicoplanin,vancomycin and linezolid were the most active agents against Staphylococcus pneumoniae(susceptible rate,100%).About 96.9%,97.8% and 98.2% Staphylococcus pneumoniae isolates were susceptible to gatifloxacin,levofloxacin,and moxifloxacin,respectively.The susceptible rates of Staphylococcus pneumoniae to ceftriaxone,chloramphenicol and amoxicillin/clavulanic acid were 81.3%,77.3%,and 68.0%,respectively.The susceptibility of Staphylococci pneumoniae to macrolides,SXT and tetracycline ranged from 11.6% to 23.6%.Conclusions The prevalence of VRE is low in China.However,methicillin-resistance among Staphylococci isolates was high.The prevalence of PNNSP isolated from (≤)3 years children is higher than in the other age population.Teicoplanin,vancomycin,and linezolid remain high activity against Staphylococci,Enterococcus faecalis,Enterococcus faecium,and Staphylococcus pneumoniae.
4.Antimicrobial resistance surveillance of gram-positive cocci isolated from 7 teaching hospitals in China in 2006
Hongli SUN ; Hui WANG ; Minjun CHEN ; Ziyong SUN ; Yunsong YU ; Bijie HU ; Yunzhuo CHU ; Jiansheng ZHUO ; Kang LIAO ; Yingchun XU ; Xiuli XIE
Chinese Journal of Laboratory Medicine 2008;31(6):635-642
Objective To investigate antimicrobial resistance among gram-positive cocci in China in 2006.Methods From Jun 2006 to Dec 2006,674 consecutive and non-repetitive gram-positive bacteria were collected from 7 teaching hospitals.The minimal inhibitory concentration(MICs)of antibacterial agents were determined by agar dilution method.Results The prevalence of penicillin.resistant(ease)and pemcllhn. intermediate S.pneumoniae(PISP)among 100 isolates was l%and 19%,respectively.Teicoplanin and vancomycin were the most active agents against S.pneumoniae.97% and 98% S.pneumoniae isolates were susceptible to levofloxacin and moxifloxacin,respectively.The susceptibility of amoxicillin/clavulanic acid, ceftriaxone and chloramphenicol are 96%,87% and 73%,respectively.The susceptible rates of penicillin. susceptible S.pneumoniae(PSSP)to cefprozil and cefaclor were 62% and 55.7%,respectively.All the PISP and PRSP isolates were resistance to the two antibiotics.The susceptibility to macrolides,trimethoprim/ sulfamethoxazole and tetracycline was lower than 35%.The prevalence of methicillin-resistant Staphylococcus aureus(MRSA)and methicillin.resistant coagulase-negative Staphylococci(MRSCON)was 48%(33%-84%)and 81%(69%-94%),respectively.The susceptible rates of MRSA to trimethoprim/ sulfamethoxazole,chloramphenicol,rifampin,and the other antibiotics in this study were 72%,66%and 45%,respectively.The susceptible rate of MRSA to marcrolides,aminoglycosides,tetracyclines and quinolones were not more than 18%.56%(30%-86%)of E.faecalis and 80%(50%.100%)of E.faecium were highly resistant to gentamicin.The susceptibility of E.faecalis to all the antibiotics except chloramphenicol and tetracycline were higher than E.faecium.All isolates of S.aureus,CoNS and E.faecalis tested were susceptible to vacomycin and teicoplanin.There were two vacomycin.resistant E.faecium strains isolated from Hangzhou.Conclusions Antimicrobial resistance patterns of gram.positive cocci differed in different regions.The resistance of gram-positive cocci to the antibiotics in this study this year was a little higher than the data of the year of 2005.Teicoplanin and vancomycin remained very high activity to gram-positive cocci.
5.Self-made pygal cloth sling for the treatment of congenital dislocation of hip in infants.
Guo-qin WANG ; Rong-jian YANG ; Xiu-xuan KANG ; Ying-hui WEN ; He-sen YUAN
China Journal of Orthopaedics and Traumatology 2011;24(9):765-767
OBJECTIVETo investigate the early clinical detection and new method for the treatment of congenital dislocation of hip in infants.
METHODSFrom 2006 to 2010, 95 infants with congenital dislocation of hip were treated with self-made pygal cloth sling, including 25 males and 70 females, with an average age of 3.2 months old ranging from 0 to 6 months. Some patients were detected incidentally for the symptoms like asymmetric muscle strength or lower limbs range of motion, and all the patients got diagnosed with dislocation.
RESULTSAfter the treatment, all of the patients received outpatient view once a month and taken X-ray examination bimonthly. Pygal cloth sling was removed after 2 months. According to the assessment criteria made by LIU Yuan-zhong, 90 patients got an excellent result, 2 good, 2 fair and 1 poor.
CONCLUSIONTreatment of congenital dislocation of hip in infants with self-made pygal cloth sling promotes the development of acetabulum and femoral head, and worthy further clinical applications.
External Fixators ; Female ; Hip Dislocation ; therapy ; Humans ; Infant ; Infant, Newborn ; Male
6.Protective effect of rapamycin on cognitive function in lupus mice by regulating Cysteine rich 61 and autophagy in hippocampus
Xiuhong PAN ; Hongli KANG ; Qiming GONG ; Yanwu YOU
Chinese Journal of Rheumatology 2022;26(6):379-386,C6-1
Objective:To investigate the effects of rapamycin (RAPA) on the cognitive function of lupus mice by regulating Cysteine rich 61 (Cyr61) and autophagy.Methods:MRL/lpr lupus mice were randomly divided into lupus group and rapamycin + lupus group, wild-type C57BL/6 mice were randomly divided into normal control group and rapamycin group with six mice in each group, RAPA + lupus group and rapamycin group were intraperitoneally injected with RAPA (2.0 mg/kg). The lupus group and the normal control group were injected with equal amounts of dimethyl sulfoxide (DMSO). Morris water maze was used to observe the cognitive function of mice. Western blotting was used to detect the expression of Cyr61, Programmed cell death-1 (Beclin-1), Microtubule associated protein 1 light chain3B (LC3B). Hematoxylin-eosin (HE) staining was used to observe the pathological changes in the hippocampus. The changes of neurons and bodies in hippocampus were observed by Nissl staining. The localization and expression of Cyr61 and LC3B in hippocampus were detected by immunofluorescence staining. One-way analysis of variance (ANOVA) was used between groups, and LSD-T test was used for pairwise comparison.Results:Western blotting results showed thatthe protein expression of Cyr61 (0.99±0.15) was significantly increased ( P=0.011), and the protein expression of Beclin-1 (0.64±0.04) and LC3B(0.54±0.05) was significantly decreased in lupus group ( P=0.025, P= 0.008) when compared with normal control group (0.73±0.08, 0.81±0.12, 0.80±0.03). The expressions of Cyr61 (0.75±0.05, 0.75±0.08), Beclin-1 (0.84±0.08, 0.92±0.04) and LC3B (0.93±0.16, 0.76±0.08) in rapamycin group and rapamycin + lupus group were not significantly changed ( P>0.05). Compared with rapamycin group, the protein expression of Cyr61 (0.99±0.15) was significantly increased ( P=0.016), and Beclin-1 (0.64±0.04), LC3B (0.54±0.05) was significantly decreased in lupus group ( P=0.013, P=0.001). The expressions of Cyr61 (0.75± 0.08), Beclin-1 (0.92±0.04) and LC3B (0.76±0.08) were not significantly changed in rapamycin+lupus group ( P=0.999, P=0.241, P=0.062). Compared with lupus group, the expression of Cyr61 (0.75±0.08) protein in rapamycin+lupus group was significantly decreased ( P=0.016), and the expression of Beclin-1 (0.92±0.04) and LC3B(0.76±0.08) protein were significantly increased ( P=0.002, P=0.017). Immunofluorescence results showed that Cyr61 and LC3B were mainly expressed in the cytoplasm of hippocampal neurons, and the quantitative detection results were consistent with western blot results, the differences were statistically significant ( P=0.025, P=0.032). HE staining showed that the levels and number of cells in the hippocampus of mice with lupus were reduced, and the arrangement was sparse, and the nuclei were hyperchromatic, showing nuclear pyknosis and migration. The results of Nissl staining showed that there were relatively fewer Nissl bodies, loose arrangement of neurons and vacuolar areas in some cells, which were improved after RAPA treatment in lupus mice. Conclusion:RAPA can protect the cognitive function of lupus mice by inhibiting the expression of Cyr61 in hippocampus and promoting autophagy.
7.In vitro expression of human factor VIII gene induced by sodium butyrate.
Jun YIN ; Hongli WANG ; Xuefeng WANG ; Haiyan CHU ; Dao LI ; Hongbing CHEN ; Qihua FU ; Baohua DUAN ; Wenying KANG ; Qiulan DING ; Zhengwu QI ; Zhenyi WANG
Chinese Journal of Hematology 2002;23(9):463-465
OBJECTIVETo explore the effect and mechanism of sodium butyrate on expression of human clotting factor VIII in vitro.
METHODSMouse NIH/3T3 cell line was transfected with recombinant plasmid vector pRC/RSV-BDD-hFVIII, which enclosed B-domain deleted (760aa approximately 1 639aa) human factor VIII cDNA (BDD-hFVIII cDNA). Then cells were incubated in Dulbecco's modification of Eagle's medium (DMEM) containing sodium butyrate for 24 hours, hFVIII: C and hFVIII: Ag in the cell culture medium were measured by ELISA assay and one-stage method, respectively. In addition, the effect of sodium butyrate on transcription of cDNA encoding the whole hFVIII, heavy and light chain of hFVIII was also investigated by means of run-on assay.
RESULTSAfter stimulation of sodium butyrate, the levels of hFVIII: C and hFVIII: Ag increased 70% than those of control. Run-on assay showed that sodium butyrate enhanced the transcription of cDNA which encoded heavy chain of hFVIII.
CONCLUSIONSodium butyrate can improve the expression of hFVIII through enhancing the transcription of hFVIII heavy chain encoding cDNA. It demonstrated that sodium butyrate had potential utility in inducing the expression of hFVIII in vitro.
3T3 Cells ; Animals ; Butyrates ; pharmacology ; Factor VIII ; genetics ; Gene Expression Regulation ; drug effects ; Humans ; Mice
8.Study on plasma coagulation factor VII (FVII) levels and polymorphisms of FVII gene in patients with coronary heart disease.
Wenying KANG ; Hongli WANG ; Lifan XIONG ; Xuefeng WANG ; Haiyan CHU ; Bin QU ; Xiangfan LIU ; Jun YIN ; Baohua DUAN ; Jinde YU ; Zhenyi WANG
Chinese Journal of Hematology 2002;23(9):457-459
OBJECTIVETo investigate the plasma levels of coagulation factor VII (FVII) and polymorphisms of FVII gene in patients with coronary heart disease (CHD), and evaluate the effect of plasma FVII levels and FVII gene polymorphisms on CHD.
METHODSPlasma FVIIa, FVII: Ag and FVIIc were measured and polymorphisms of FVII gene were analyzed in 149 control cases and 60 CHD cases, including 33 acute myocardial infarction (AMI) cases by a combination of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and agarose gel electrophoresis.
RESULTSFVIIa, FVIIc in AMI group were significantly higher than that in control group, but FVII: Ag wasn't. There were no significant difference in plasma FVIIa, FVII: Ag and FVIIc between CHD and control group. The IVS7 genotypic frequency in AMI group was significantly different from that in control group. There was no significant difference in genotypic frequencies and allelic frequencies in other polymphism sites. FVII: Ag was significantly higher in -402A homozygote than that in -402G homozygote.
CONCLUSIONSIncreased FVII levels, especially FVIIa and FVIIc in plasma, may contribute to coronary artery thrombosis. There was significant difference in IVS7 genotype frequency between control and AMI groups, but the rest weren't significantly different. FVII: Ag was significantly higher in -402A homozygote than that in -402G homozygote. Polymorphism of -402 G/A may play an indirect role in AMI by regulating plasma FVII levels.
Coronary Disease ; blood ; genetics ; Factor VII ; analysis ; genetics ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length
9.Preliminary experimental research on gene therapy for hemophilia A.
Jun YIN ; Hongli WANG ; Yiqun HU ; Xuefeng WANG ; Bin QU ; Haiyan CHU ; Baohua DUAN ; Wenying KANG ; Zhengwu QI ; Zhenyi WANG
Chinese Journal of Hematology 2002;23(3):138-142
OBJECTIVETo accomplish a kind of therapeutic gene for hemophilia A, and observe the expression of human factor VIII (hF VIII) in vivo.
METHODSHuman clotting factor VIII cDNA with B-domain deleted (Delta760aa approximately 1639aa) was inserted into vector pRC/RSV to form pRC/RSV-hF VIII BD, which conjugated with in vivo liposome transfection reagent (DOTAP-Cholesterol) to accomplish a kind of therapeutic gene, pRC/RSV-hF VIII BD-DOTAP-Cholesterol. Mice were injected with pRC/RSV-hF VIII BD-DOTAP-Cholesterol i.m. and sacrificed 48 hours, 10 days, 20 days, 30 days, 40 days and 50 days later, respectively. Tissues such as heart, liver, spleen, lung, kidney and muscle were harvested, the distribution and transcription as well as expression of hF VIII BD cDNA were detected by means of PCR, RT-PCR and immunohistochemistry techniques. In addition, the antigen and antibody of hF VIII in plasma were measured.
RESULTSThere was high expression of hF VIII in plasma and tissues at the 48(th) hour after injection. On day 10, antigen level of hF VIII in plasma reached its peak, 17.55 ng/ml, and gradually reduced later. The antibody of hF VIII in plasma emerged on day 10 after injection, and increased and gradually reached 37.06 U/ml on day 50 after injection. PCR, RT-PCR and immunohistochemistry showed that hF VIII BD cDNA and its transcription as well as expression existed in all kinds of tissues, and lasted longer in spleen, lungs and kidneys than in heart, liver and muscle.
CONCLUSIONTherapeutic gene, pRC/RSV-hF VIII BD-DOTAP-Cholesterol, produced by combination of pRC/RSV-hF VIII BD and DOTAP-Cholesterol liposome can express human F VIII successfully in vivo, which lays an experimental foundation for curing hemophilia A by gene-drug in clinic.
Animals ; DNA, Complementary ; Disease Models, Animal ; Factor VIII ; biosynthesis ; genetics ; therapeutic use ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Hemophilia A ; therapy ; Humans ; Liposomes ; Mice ; Mice, Inbred BALB C ; Tissue Distribution ; Transfection
10.Molecular analysis of two pedigrees with hereditary F VII deficiency.
Haiyan CHU ; Hongli WANG ; Xuefeng WANG ; Xuemei GUO ; Bin QU ; Baohua DUAN ; Jun YIN ; Wenying KANG ; Zhenyi WANG
Chinese Journal of Hematology 2002;23(3):130-133
OBJECTIVETo identify the mutation of coagulation factor VII (F VII) gene in two pedigrees with hereditary F VII deficiency.
METHODSF VII gene mutations were analysed in two propositi and their family members by direct DNA sequencing. Allele specific PCR and PCR combined with restricted enzyme digestion were used to confirm the detected mutations.
RESULTSTwo gene mutations were detected in the propositus of pedigree A: G to C transition at position 6390 resulting in Trp40Cys and G to A at 11496 resulting in Arg353Gln, both are heterozygotes. The heterozygosity for polymorphism Arg353Gln was confirmed with the restriction enzyme Msp I digestion in his mother. In the propositus of pedigree B, there was a T to G transition at position 11482 resulting in His348Gln, heterozygosity of which was confirmed with Nsp I digestion in the propositus and his daughter. G to T transition at position 11514 resulting in Thr359Met was also found in the propositus of pedigree B, and the heterozygosity for Thr359Met was confirmed with allele specific PCR in the propositus and his son.
CONCLUSIONThree missense mutations were found in two pedigrees with hereditary F VII deficiency. A novel Trp40Cys mutation was reported for the first time.
Factor VII ; genetics ; Factor VII Deficiency ; genetics ; Female ; Heterozygote ; Humans ; Male ; Mutation, Missense ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; methods