1.Investigation of surfactant protein-C gene on respiratory distress syndrome in the Han nationality new-borns of the Inner Mongolia
Hua MEI ; Yuheng ZHANG ; Dan SONG ; Ya'nan HU ; Rong HONG ;
Chinese Pediatric Emergency Medicine 2015;22(7):454-457
Objective To investigate the relationship between single nucleotide polymorphisms of surfactant protein C(SP-C)gene and respiratory distress syndrome(RDS)in the Han nationality newborns in Inner Mongolia and whether there is a mutation occurs on SP-C gene exon 4 and 5.Methods One hundred newborns with RDS(case group)and 100 newborns without RDS(control group)were selected.PCR gene analysis was used to establish the genotype and allele frequencies of exon 4 (T138N)and 5 (S186N)on SP-C.Results In the Han nationality newborns of Inner Mongolia region,there was no mutation on SP-C gene exon 4 and 5.Exon 4(T138N)on SP-C could be checked out three genotypes:namely AA,AC and CC.The genetic polymorphisms of exon 4 on SP-C were not statistically different between the case group and the control group(χ2 ﹦0.744,P ﹦0.689).Besides,exon 5(S186N)on SP-C could also be checked out three genotypes:namely AA,AG and GG.The genetic polymorphisms of exon 5 on SP-C were also not statistically different between the case group and the control group(χ2 ﹦0.770,P ﹦0.681 ).Conclusion There is no mutation on SP-C gene exon 4 and 5.The genetic polymorphism of exon 4 and 5 on SP-C displays no signifi-cant correlation with RDS of the Han nationality newborns in Inner Mongolia.
2.Ciprofloxacin non-susceptible Streptococcus pyogenes due to mutations in parC gene and its homology analysis
Xiaofei ZHANG ; Yunjian HU ; Dan CHI ; Yanyan HU ; Hongwei ZHOU ; Rong ZHANG
Chinese Journal of Laboratory Medicine 2013;(5):425-429
Objective To investigate the resistant mechanism of Streptococcus pyogenes to ciprofloxacin and its homology.Methods Forty-eight isolates of Streptococcus pyogenes were collected from patients diagnosed with scarflet fever in districts of Beijing in March,2012 and MIC to ciprofloxacin and other 7 common antibiotics in clinic were detected by using blood M-H agar dilution method.Thirteen isolates,which have MICs≥4 mg/L against ciprofloxacin,were detected for mutations of Fluoroquinolone resistance genes gyrA,gyrB,parC,parE.At the same time,4 isolates,with MIC ≤ 0.25 mg/L against ciprofloxacin,were used for comparison.Homology analysis of 17 isolates from different areas of Beijing was performed by using the method of pulsed field gel electrophoresis.Results Sensitive rates of Streptococcus pyogenes to levofloxacin,ampicillin and penicillin were all 100%.The resistance rates to tetracycline,erythromycin and clindamycin were 91.7% (44/48),91.7% (44/48) and 89.6% (43/48),respectively.MIC50 of ciprofloxacin,levofloxacin and moxifloxacin was 2 mg/L,1 mg/L and ≤ 0.25 mg/L,respectively ; MIC90 was 4 mg/L,2 mg/L and 0.5 mg/L,respectively.Of the 48 isolates of Streptococcus pyogenes,12 isolates showed the MIC at 4 mg/L,while one isolate has a MIC against ciprofloxacin at 8 mg/L,which isolated from Chaoyang district.Analysis of sequence of chromosome mediated fluoroquinolone resistance genes in those 13 ciprofloxacin non-susceptible isolates exhibited that there were 12 isolates that harbored Ser79Phe/Tyr mutation and 10 isolates harbored Ala121Val in parC gene.It is shown that one isolate contained Ser79Phe mutation in parC gene in the occurring of Ser371Leu mutation in parE gene for the first time,but there was no marked increase in ciprofloxacin MIC (MIC =4 mg/L).There were no mutations in gyrA and gyrB genes.The PFGE results demonstrated that the 17 tested isolates could be divided into 7 clones.The clone A isolates from Chaoyang,Daxing,Fengtai,Shunyi and Shijingshan district have a MIC ≥ 4 mg/L against ciprofloxacin,which covered 69.2% of all MIC ≥4 mg/L isolates.The clone C isolates from Huairou district were MIC ≥4 mg/L isolates.B,D,E,F and G clones isolates come from different districts.Conclusions The mutation of parC gene was the main reason that contribute to the slightly increase of ciprofloxacin MIC in Streptococcus pyogenes isolated from Beijing.The PFGE analysis showed that there was a small scale prevalence caused by the infection of Streptococcus pyogenes in some districts.
3.Molecular types of carbapenem-non-susceptible Escherichia coli isolates and their mechanism of drug resistance
Rong ZHANG ; Dan CHI ; Jiachang CAI ; Yanyan HU ; Hongwei ZHOU ; Wei YANG ; Huoyang LYU ; Gongxiang CHEN
Chinese Journal of Microbiology and Immunology 2014;(4):256-263
Objective To investigate the molecular types of carbapenem-non-susceptible Esche-richia coli ( E.coli) isolates and their mechanism of carbapenem resistance .Methods Twenty-two carbap-enem-non-susceptible E.coli strains were isolated from 3 hospitals in Hangzhou from 2007 to 2011.The mini-mum inhibitory concentrations ( MICs) of antimicrobials to those isolates were determined by agar dilution method and E-test.The molecular mechanisms of carbapenem resistance of E.coli isolates were analyzed by conjugation experiment,PCR and DNA sequencing.Pulsed-field gel electrophoresis (PFGE),multilocus se-quence typing ( MLST ) , and phylogenetic typing were performed to analyze the molecular epidemiology of those isolates.Results The MICs of imipenem and meropenem to 22 E.coli isolates were ranged from 1 μg/ml to 16 μg/ml,and the MICs of ertapenem were 2 μg/ml to 64 μg/ml.All E.coli isolates produced the KPC-2 carbapenemase and various β-lactamases , and some of them also produced plasmid-mediated AmpC enzymes.Carbapenem resistance was transferred by conjugation and transformation from 22 E.coli iso-lates to E.coli EC600 strains.The E.coli transconjugants or transformants acquired the blaKPC-2 gene and showed similar antibiotic susceptibility patterns in comparison with donor strains .Only a few isolates were in-distinguishable or closely related as indicated by PFGE .Four sequence types including ST131 (9 isolates), ST648 (5 isolates),ST38 (2 isolates) and ST405 (2 isolates) were identified by MLST.Phylogenetic analy-sis indicated that 9 ST131 isolates belonged to phylogenetic group B 2 and the other isolates belonged to group D (11 isolates),group B1 (1 isolate) and group A (1 isolate),respectively.Conclusion The sequence type of prevalent E.coli isolates producing KPC-2 from Hangzhou was ST131,which is an international epi-demic,multidrug-resistant clone,followed by ST648.
4.Pre-treatment with melatonin inhibits oleic acid-induced acute lung injury in rats
Qun WANG ; Qing TIAN ; Rong LIU ; Zhuangzhi LIU ; Maoqiong HU ; Dan KE ; Shaohua ZHANG ; Xiaochuan WANG
Chinese Journal of Pathophysiology 2007;23(4):767-771
AIM:To assess the protective role of melatonin(MEL)in a rat model of oleic-induced acute lung injury.METHODS:Twenty-four rats were randomly allocated to three groups as follows:saline(NS)injection group,oleic acid(OA)injection group and MEL plus OA injection group,the lavage protein,lung wet-to-dry weight ratio,malondialdehyde(MDA)content,superoxide dismutase(SOD)activity and lung histopathology were examined.RESULTS:(1)Injection 0.15 mL/kg of OA led to a severe acute lung injury(ALI),characterized by significantly increasing in lavage protein,lung coefficient(P<0.01),and by histopathological alterations which presented hemorrhage,edema.thickened alveolar septum and the existence of inflammatory cells in alveolar spaces;(2)Infusion of MEL(20 mg/kg,intraperitoneally for 60 min before the oleic acid)markedly alleviated above-mentioned symptom induced by OA,consistent with decrease of MDA level(P<0.01) and the increase of SOD activty(P<0.01).CONCLUSION:Pre-treatment with MEL can attenuate the OA -induced ALI in rats via cleaning and preventing the formation of free radicals and further lessening the increase of alveolocapillary membrane permeability,these data suggest that MEL may be effective in the prevention of ALI.
5.Inhibitory effects of RNA interference on expression of matrix metalloproteinase-9 gene and invasiveness and adhesion in ovarian cancer cells
Xiao-Xia HU ; Li LI ; Dan-Rong LI ; Wei ZHANG ; Bu-Jian TANG ;
Chinese Journal of Obstetrics and Gynecology 2000;0(09):-
Objective To investigate the inhibitory effects of RNA interference(RNAi)on the expression of matrix metalloproteinase-9(MMP-9)gene and invasiveness and adhesion of ovarian cancer cells.Methods Four groups of different specific target sequence in coding region of MMP-9 and one non- specific sequence were chosen,which were Sitel,Site2,Site3,Site4 and Site5.Small interference RNA (siRNA)expression cassettes(SEC)were constructed by PCR and transfected into ovarian cancer HO- 8910PM cells.RT-PCR and western blot were used to detect mRNA and protein expression of MMP-9 gene; the abilities of invasion and adhesion were detected by Matrigel invasion assay and cell adhesion assay. Results The expression of MMP-9 was inhibited and the inhibitory effects of different sequence were varied.The mRNA expression was 0.64?0.06,0.47?0.07,0.55?0.10 in Sitel,Site2,Site3 group, and protein expression was 0.30?0.09,0.27?0.08,0.37?0.12,respectively.Site2 group had the most efficient inhibitory effect,followed by Sitel and Site3 groups.Cell growth curve revealed that cell growth was significantly inhibited in Site2 group.Invasiveness and adhesion were significantly reduced,the inhibitory rate on invasion in Site1,Site2,Site3 groups were 50.0%,50.0% and 37.5%,respectively;the inhibitory rate on adhesion in Site1,Site2,Site3,Site4 groups were 43.8%,48.8%,33.9%,24.2% at 60 min and 41.6%,40.2%,35.1%,16.0% at 90 min,respectively.Conclusions RNAi exists in ovarian HO-8910PM cells.MMP-9 siRNA can specifically down-regulate MMP-9 expression and lead to the inhibition of invasiveness and adhesion in ovarian cancer cells.
6.The assessment and comprehensive nursing interference of pain after thoracic surgery
Jihong ZHONG ; Rong HU ; Xuemin ZHU ; Xia ZHANG ; Yulian SUN ; Dan WU ; Juan YU ; Xin ZHOU
Journal of Medical Postgraduates 2015;(2):189-191
Objective Because of the major trauma from thoracotomy , postoperative pain may arouse patients'psychological and physiological stress response and hence affected the treatment outcome and functional recovery seriously .We retrospectively studied the correlation between the staging of pain and nursing interference to investigate the effect of nursing interference on the pain intensity after thoracic surgery . Methods Five hundred and eighty cases of patients surviving thoracotomy between December 2013 and March 2014 in Nanjing Jingling Hospital were reviewed .Correlations between comfortable nursing measures such as effective analgesia , postural care , catheter care , environmental interventions , psychological intervention and the standard assessment of pain were analysed according to postoperative pain stage . Results With comprehensive nursing interference , the highest pain score occurred in the first 24 hour (2.89 ±0.39).The score was reduced gradually to 2.25 ±0.90 in stage Ⅱ, 1.58 ±0.57 in stage Ⅲ and 1.06 ±0.24 in stage Ⅳrespectively . Conclusion Comprehensive nursing interference according to pain staging may relieve pain effectively after thoracotomy .
7.Peptide CY11 conjugated polyethylenimine-beta-cyclodextrin for gene delivery.
Yi-Ping HU ; Qi-Ying JIANG ; Dan CHEN ; Dan-Jun ZHAO ; Xiao LU ; Hai YU ; Qing-Qing WANG ; Jun ZHOU ; Xiu-Rong HU ; Gu-Ping TANG
Journal of Zhejiang University. Medical sciences 2009;38(1):24-30
OBJECTIVETo develop a novel non-viral gene delivery vector CY11-PEI-beta-CyD and to test its gene transfection efficiency.
METHODSCY11 (CGMQLPLATWY) was conjugated to polyethylenimine-beta-cyclodextrin to form CY11-PEI-beta-CyD with a cross-linker [N-succinimidy-3-(2-pyridyldithio) propionate, SPDP]. (1)H-NMR and TGA were used to confirm the structure of vector. The DNA condensing ability of CY11-PEI-beta-CyD was investigated by gel retardation assay. Cytotoxicity of CY11-PEI-beta-CyD was determined by MTT assay and transfection efficiency was investigated in COS-7, Hela and B16 cells.
RESULTCY11 was conjugated onto PEI-beta-CyD successfully, confirmed by(1)H NMR and TGA. The novel vector effectively condensed DNA at N/P ratio of 4îIt showed low cytotoxicity up to the concentration was 160 Mgr;g/ml. The transfection efficiency was 17-fold higher than that of PEI 25 kDa at N/P ratio of 20.
CONCLUSIONThe novel vector CY11 -PEI-beta-CyD with low cytotoxic and high transfection efficiency may be used as a potential carrier for gene delivery.
Cell Line ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Humans ; Peptide Fragments ; chemistry ; Polyethyleneimine ; chemistry ; Receptors, Fibroblast Growth Factor ; chemistry ; beta-Cyclodextrins ; chemistry
8.Micromeritic evaluation of the direct compression excipient LubriTose AN.
Yi-Lan ZHANG ; Chao TIAN ; Dan-Rong HU ; Xue KE ; Ji-Lai TIAN
Acta Pharmaceutica Sinica 2012;47(5):640-645
This study is to report the evaluation of the micromeritic properties of LubriTose AN, which is expected to provide preliminary theoretical basis for the direct compression technology. From the aspects of flowability, compressibility and dilution potential, the angle of repose, flow velocity, the Carr' index, tensile strength, elastic recovery, yield pressure and the lubricating ability of LubriTose AN were determined. Also, model drugs were selected to investigate the dilute potential under the desirable compressing performance. Compared to the physical mixtures, the flowability of LubriTose AN was better, and the deformation mechanism was the same with anhydrous lactose, both brittle deformation. The compressibility and compaction of LubriTose AN was slightly better than that of physical mixtures under low and moderate pressure. The dilution potential of LubriTose AN were high for most of hydrophobic drugs. The lubricate ability was desirable under different rotational speeds. LubriTose AN is an excellent co-processed excipient, which is helpful for the promotion and improvement of the tablet manufacturing level.
Drug Compounding
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Elasticity
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Excipients
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chemistry
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Glycerides
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chemistry
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Ibuprofen
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administration & dosage
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chemistry
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Lactose
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chemistry
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Lubricants
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chemistry
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Lubrication
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Particle Size
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Pressure
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Technology, Pharmaceutical
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methods
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Tensile Strength
9.Preparation and assessment of heterotopic osteoinduction of beta-TCP/rhBMP-2 composite.
Dan WANG ; Yunyu HU ; Guangyue ZHAO ; Rong LU ; Guang YANG ; Changqiong ZHENG ; Kenan XIE
Chinese Journal of Traumatology 1999;2(1):13-16
OBJECTIVE: To search for ideal bone graft substitute. METHODS: The beta TCP/rhBMP-2 composite was constructed by combining beta-Tricalcium phosphat (beta-TCP) that was prepared by the authors with recombinant human morphogenetic protein-2 (rhBMP-2) and was implanted into the muscle pouches in the thigh of mice. beta-TCP alone was implanted on the opposite side as controls. At intervals of 1,3,7,14 and 28 days after the implantation, the specimens were obtained, and histologic study and alkaline phosphatase assay (7,14,28 days) were performed. RESULTS: There was a large amount of cartilage and bone formation within the composite, increasing with time; whereas there was no new bone formation where beta-TCP alone was implanted. Besides, the levels of alkaline phosphatase in the beta-TCP/rhBMP-2 implants also were increasing with time and were higher than those in controls. CONCLUSIONS: The results indicate that beta-TCP/rhBMP-2 composite possesses heterotopic osteoinductive potential.
10.Methylation in Promoter Region of SLC6A2 Gene in Heart Failure Patients and Its Correlation with Qi Deficiency/Blood Stasis Syndrome.
Jie-wei LUO ; Xiao-rong MENG ; Fang-meng HUANG ; Dan HU ; Xiao YANG ; Xing-yu ZHENG ; Shi-chao WEI ; Miao-miao GUO ; Shi-ming WU ; Hong-yi CHEN
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(12):1448-1454
OBJECTIVETo explore the methylation status in promoter region of norepinephrine transporter gene (NET, SLC6A2) in heart failure ( HF) patients and its correlation with qi deficiency/blood stasis syndrome (QDS/BSS).
METHODSThirty-six patients with heart failure (NYHA classification III to IV) were recruited in the study (as the heart failure group) and their scores of QDS/BSS were evaluated. Besides, a healthy elderly group (30 cases) and a healthy youth group (30 cases) were also set up. They were recruited from Physical Examination Center of Fujian Provincial Hospital. Pyrosequencing was applied to detect the methylation in promoter region of SLC6A2 gene, and the total methylation index (MTI) of CpG island was calculated. The correlation between the methylation status in promoter region of SLC6A2 and scores of QDS/BSS was assessed using Pearson and Partial analyses. Risk factors were screened and adjusted using Logistic regression.
RESULTSBy one-factor analysis of variance, the total MTI in the HF group (219.72% ± 54.03%) was obviously higher than that in the healthy elderly group (194.47% ± 34.92%) and the healthy youth group (161.60% ± 41.11%) (all P < 0.05). Meanwhile, the total MTI was higher in the healthy elderly group than in the healthy youth group (P < 0.01). By covariance analysis , after controlling age and BMI, the total MTI was higher in the HF group than in the healthy elderly group (P = 0.041), while it was higher in the healthy elderly group than in the healthy youth group (P = 0.016). Age was found to play an essential role in affecting MTI of SLC6A2 gene promoter region among the 3 groups (F = 16.447, P = 0.01). The total MTI was quite lower in the healthy youth group. Results of Partial correlation analysis showed MTI was positively correlated with scores of qi deficiency and blood stasis respectively (r = 0.494 and 0.419 respectively, both P < 0.05). Logistic regression analysis showed after adjusting confounding factors, the relative risk (OR value) of total MTI of SLC6A2 gene in promoter region was 1.038 (95% CI, 1.006 to 1.071, P = 0.020).
CONCLUSIONSAbnormally elevated methylation of the promoter region of SLC6A2 gene is one of risk factors for HF. In addition, the degree of methylation of the promoter region of SLC6A2 gene was positively correlated with the severity of QDS/BSS.
Adolescent ; Aged ; DNA Methylation ; Heart Failure ; genetics ; physiopathology ; Humans ; Logistic Models ; Medicine, Chinese Traditional ; Norepinephrine Plasma Membrane Transport Proteins ; genetics ; Promoter Regions, Genetic ; Qi