1.Significance of aberrant expression of DNA methyltransferase 1 in cervical cancerization tissue and cervical cancer cell lines
Jintao WANG ; Ling DING ; Junxia HAO ; Weimin ZHAO ; Qin ZHOU ; Min HAO ; Shuli SHAO
Cancer Research and Clinic 2012;(12):797-800
Objective To explore the effects of aberrant expression of DNA methyltransferase 1 (DNMT1) in cervical cancerization tissue and cervical cancer cells.Methods Cervical tissues were collected from 80 cases with a diagnosis of invasive cervix squamous cell carcinoma (SCC),53 cases with high-grade cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ),52 cases with low-grade cervical intraepithelial neoplasia (CIN Ⅰ)and 53 cases with normal cervix (NC).Meanwhile,Caski (HPV16-positive) and C33A (HPV-negative) cells selected from cervical cancer cell lines were cultured routinely in vitro.The expression of DNMT1 protein and mRNA were examined by Western blot analysis and real-time PCR (qRT-PCR) in the tissues and cells,respectively.Results The levels of DNMT1 protein were 1.33,1.84 and 2.28,and the Ct-ratios (DNMT1/β-actin) of DNMT1 mRNA were 1.27,1.27 and 1.26 in CIN Ⅰ,CIN Ⅱ/Ⅲand SCC group,respectively.Comparing with NC group,the expression of DNMT1 protein or mRNA was elevated in deficient cervical groups,with statistical significance (F =110.57,P < 0.001,F =2.68,P =0.048).The expression levels of DNMT1 protein were increased steadily according to severity of the cervix lesions (x2tend =50.80,P < 0.001),however,the expression of DNMT1 mRNA was not observed the same tendency (x2tend =3.63,P > 0.05).The results from experiment in vitro showed that the levels of DNMT1 protein or mRNA were both higher in Caski cell than in C33A cell,especially for DNMT1 mRNA with significantly difference (t =7.134,P =0.002).Conclusion Aberrant expression of DNMT1 protein or mRNA could link with the risk of cervical cancerization by both transcriptional and posttranscriptional mechanisms.There would be a synergistic effect between overexpression of DNMT1 and HPV16 infection in the progression of cervix carcinogenesis.
2.Effects of indomethacin on human choroidal melanoma OCM-1 xenografts in nude mouse
Xin-rui, LUO ; Hao, CHEN ; Qin-xiang, ZHENG ; Ling, QIN ; Min, LI ; Wen-sheng, LI
Chinese Journal of Experimental Ophthalmology 2011;29(1):8-12
Background Choroidal melanoma(CM)is a common form of primary ocular cancer in adults.It is reported that indomethacin has inhibitory effect on many tumor in vitro and in vivo,but whether it can inhibit the growth of CM has not been published. Objective This study was to investigate the anti-tumor activity of indomethacin on the growth of human CM OCM-1 cell xenografts in nude mice. Methods OCM-1 cells were subcutaneously implanted on 24 SPF female BALB/C.nu/nu nude mice to establish ectopic models of human CM.The nude mice with the tumor 5 mm were randomly divided into 4 groups:untreated group,normal saline solution(NS) group,indomethacin 1 ms/kg group,indomethacin 2 ms/kg group.The 1 mS/kg or 2 ms/kg indomethacin was intraperitoneally injected for 14 consecutive days in indomethacin 1 ms/kg group and indomethacin 2 me/kg group respectively.and 0.2 ml of 2%NS-DMSO was used at a same way in the NS group.No any agent was used as the untreated group.The volume and weight of implanted tumor as well as inhibitory rates of indomethaein on tumor were calculated.The expression of ki67 and survivin proteins were measured with immunohistochemistry,and the expression of survivin mRNA in CM was assessed by RT-PCR. ResuIts The tumor of indomethacin treatment group was reduced in volume and weight with a significant difference between treatment group and control group as well as indomethacin 1 ms/ks group and indomethacin 2 ms/kg group(P<0.05).The inhibitory rate of indomethacin 1 ms/kg and 2 ms/kg for tumor was 22.86%,48.00%respectively.The prolifiration index (PI)of ki67 in these 4 groups were (76.73±3.34)%,(73.30±2.95)%,(55.97±2.24)%,(32.87±2.91)%respectively,and significant difference was found in PI between indomethacin 2 mg/kg group and untreated group or NS group(P<0.05),but there was not significant difference between indomethacin 1 mg/kg and 2 ms/kg group(P>0.05).Compared with the control group,the indomethacin treatment groups showed the decreased expression of survivin protein and mRNA,and significant difference was found between indomethaein 2 ms/kg group and untreated group or NS group(P<0.05),however,no significant difference was found between indomethacin 1 mg/kg and 2 mg/kg group(P>0.05). Conclusion Indomethacin inhibits the growth of CM in nude mice through inhibiting the expression of survivin in the tumor and accelerating cell apoptosis and inhibiting tumor cell proliferation.
3.bla_(LAP-2),A Novel Subtype of ?-Lactamases Gene in Enterobacter cloacae in China
Zhimi HUANG ; Zuhuang MI ; Qiuju CHU ; Hao SHAN ; Shouhui XIA ; Weiping YANG ; Haiyan YANG ; Ling QIN
Chinese Journal of Nosocomiology 2006;0(05):-
OBJECTIVE To investigate the resistance genes in HZB9055 strain of Enterobacter cloacae isolated from the PLA 98th Hospital,Huzhou District,Zhejiang Province,China.METHODS HZB9055 Strain of E.cloacae was isolated from the inpatient in May,2004,41 kinds(or groups) of genes were analyzed by PCR and verified by DNA sequencing.RESULTS In HZB9055 strain of E.cloacae,3 kinds of genes were positive including blaLAP,blaMIR and qnrS.The 38 kinds of rest genes were all tested negative.The blaLAP gene sequence including 858 nucleotides,which had the amino acid mutation in position 193 compared with LAP-1 type narrow-spectrum ?-lactamase(GenBank accession number:EF026092),and was nominated LAP-2(GenBank accession number:EU159120).CONCLUSIONS At least 3 kinds of resistance genes exist in HZB9055 strain of E.cloacae,and blaLAP-2 is a novel subtype of ?-lactamases gene.
4.Antimicrobial-resistant Genes and Consanguinity in Multi-resistant Pseudomonas aeruginosa
Zhimi HUANG ; Hao SHAN ; Manying GUO ; Yu CHEN ; Lei WU ; Zuhuang MI ; Qingyun ZHUGE ; Ling QIN
Chinese Journal of Nosocomiology 1994;0(04):-
OBJECTIVE To investigate the antimicrobial-resistant genes and consanguinity in multi-resistant Pseudomonas aeruginosa(MRPA) isolated from the 98th Hospital of PLA,Huzhou,Zhejiang Province,China.METHODS Thirty strains of MRPA were isolated from hospitalized patients between Sep 2003 and Oct 2004.Twenty four kinds of genes of blaTEM,blaSHV,blaOXA-10 group,blaPER,blaVEB,blaIMP,blaVIM,blaGES,blaCARB,blaDHA,blaMIR,aac(3)-Ⅰ,aac(3)-Ⅱ,aac(3)-Ⅲ,aac(3)-Ⅳ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ,aph(3′)-Ⅵ,oprD,qacE△1-sul1,catB,and cml1 were analyzed by PCR and verified by DNA sequencing.Resistant-genes cluster analysis was performed by Average.RESULTS In 30 strains of MRPA the positive rate of genes of blaTEM,blaOXA-10 group,blaCARB,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ,qacE△1-sul1,and cml1 were 66.7%,3.3%,3.3%,76.7%,3.3%,33.3%,53.3%,26.7%,83.3%,and 3.3%,respectively,and the deficiency rate of oprD gene was 90.0%.The gene of blaOXA-10 group was sequenced and determined to be blaOXA-10 subtype ESBL gene.But the rest of genes were all negative.According to the cluster analysis of resistant-gene,30 strains of MRPA isolated could be classified into four subgroups,which were caused by the infection in hospital.CONCLUSIONS At least 10 kinds of antimicrobial-resistant genes exist in MRPA isolates,and the deficiency rate of oprD gene is very high.MRPA can induce clone transmitted hospital infection and it has fulminant prevalence.
5.?-Lactamase Genes in Four Kinds of Nonfermenting Gram-negative Bacilli
Jianzhong CHEN ; Zhimi HUANG ; Hao SHAN ; Yu CHEN ; Lei WU ; Jing WU ; Zuhuang MI ; Ling QIN
Chinese Journal of Nosocomiology 2006;0(08):-
OBJECTIVE To investigate the ?-lactamase genes in 4 kinds of nonfermenting Gram-negative bacilli isolated from the 98th Hospital of PLA. METHODS Sixty strains of Acinetobacter baumannii,30 strains of Pseudomonas aeruginosa,19 strains of Stenotrophomonas maltophilia,and 15 strains of Flavobacterium were isolated from hospitalized patients.Nine kinds of ?-lactamases genes of TEM,SHV,OXA,CTX-M,PER,VEB,IMP,VIM and GES were analyzed by PCR and verified by DNA sequencing. RESULTS In A.baumannii and(P.aeruginosa),the positive rates of gene of TEM were 100.0% and 66.7%,respectively.SHV gene was positive in 18 of 60 strains of A.baumannii tested,17 of which were SHV-12 subtype ESBLs.The other was a new SHV type ?-lactamase nominated SHV-48.OXA gene was positive in 1 of 30 strains of P.aeruginosa tested,it was an OXA-10 subtype ESBLs.But the rest of genes were all negative. CONCLUSIONS There exist 4 kinds of(?-lactamase) genes at least in nonfermenting Gram-negative bacilli including TEM-1,SHV-12,SHV-48,and(OXA-10.)
6.Effect of water-soluble polymers on the inhibition of osthole crystallization.
Acta Pharmaceutica Sinica 2010;45(12):1559-1564
This paper is to study the inhibitory effect of water soluble polymers--methyl cellulose (MC), hydroxypropyl methyl cellulose (HPMC), hydroxypropyl cellulose (HPC-M), poloxamer (F68) and polyvidon (PVP) on osthole (OST) crystallization and investigate the impact of polymer concentration and viscosity on crystallization behavior. Also, UV spectrophotometry method was used to determine the drug concentration at different time point to draw the OST concentration-time curve. Results show that HPMC has the most significant inhibition effect on OST crystallization, and drug concentration level is 1.61 times higher than that in control solution within 8 h followed by PVP (1.54) and MC (1.45) respectively. The kinetics of OST recrystallization can be described using first-order reaction, and the crystallization rate constants obtained by analyzing the regression equation indicate that HPMC-60SH-4000 and HPMC-60SH-10000 can greatly influence OST crystal formation. The dissolution rate of drugs precipitated from water-soluble polymer solutions is faster compared with controls in pH 1.2 HCl and pH 6.8 phosphate buffers, which demonstrated that water-soluble polymers can not only change the behavior of drug crystallization but markedly improve the dissolution rate of water insoluble drugs.
Cellulose
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analogs & derivatives
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chemistry
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Cnidium
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chemistry
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Coumarins
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chemistry
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isolation & purification
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Crystallization
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Hypromellose Derivatives
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Kinetics
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Methylcellulose
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analogs & derivatives
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chemistry
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Plants, Medicinal
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chemistry
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Poloxamer
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chemistry
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Polymers
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chemistry
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Povidone
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chemistry
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Viscosity
7.Preparation and evaluation of clarithromycin emulsion for injection.
Acta Pharmaceutica Sinica 2006;41(10):945-949
AIMTo prepare clarithromycin emulsion and investigate its pharmacokinetics in rats. And to do irritation test of the emulsion.
METHODSHigh pressure homogenization method was used to prepare clarithromycin emulsion, and the Nicomp380 machine was used to test the mean particle size and zeta-potential of clarithromycin emulsion. Irritation of emulsion was also evaluated compared with the positive control of clarithromycin solution using rat paw lick test and rabbit ear vein irritation test. Microbiological assay method was used for determining the drug concentration in plasma. Pharmacokinetics of two dosage forms in rats was also studied.
RESULTSThe mean particle size and zeta potential of clarithromycin emulsion were 156 nm and -31.8 mV, respectively. The emulsion was stable during the storage time at 4 degrees C for 6 month. The pain caused by emulsion reduced significantly compared with that of clarithromycin solution based on the results of rat paw lick test and rabbit ear vein test. The drug concentration-time curves of clarithromycin emulsion and clarithromycin solution were similar and could be described by two compartment model. AUC(0-1) of clarithromycin emulsion and clarithromycin solution were (66.76 +/- 16.34) and (59.00 +/- 11.20) microg x h x mL(-1), respectively.
CONCLUSIONStable emulsion could be prepared using high pressure homogenization method, and irritation caused by i.v. injection could be reduced significantly by using clarithromycin emulsion.
Animals ; Anti-Bacterial Agents ; administration & dosage ; adverse effects ; pharmacokinetics ; Area Under Curve ; Clarithromycin ; adverse effects ; blood ; pharmacokinetics ; Drug Compounding ; methods ; Emulsions ; Female ; Injections ; Male ; Pain ; chemically induced ; Pain Measurement ; methods ; Particle Size ; Rabbits ; Rats ; Rats, Wistar ; Solutions
8.Hearing in Noise Test for OIder AduIts with Age-reIated Hearing Loss
Jingwen SUN ; Zhiwu HUANG ; Jianyong CHEN ; Ling MEI ; Yun LI ; Yan REN ; Min SHEN ; Qin ZHANG ; Hao WU
Journal of Audiology and Speech Pathology 2015;(1):16-20
Objective To evaluate the speech recognition ability in noise enviroment in older adults with age-related hearing loss and to discuss the influences of age ,hearing loss and cognitive function on the ability of speech recognition in noise enviroment .Methods A total of 90 older adults aged over 60 years at the hearing clinic of Xinhua hospital were divided into three groups according to the age ,which were 60~69 y group (88 ears) ,70~79 y group (64 ears) ,> 79 y group (28 ears) .All the participants completed pure tone audiometry ,the Mandarin hearing in noise test (MHINT) and the mini-mental state examination (MMSE) .The pure tone average hearing threshold (PTA) ,signal-to -noise ratio (SNR) of MHINT and MMSE scores were obtained .We used the statis_tical software SPSS 17 .0 to analyze the correlations among age ,PTA ,adaptive SNR and MMSE scores .ResuIts ①PTA and SNR increased with the age .The difference of PTA and SNR for different age groups was statistically sig_nificant (P<0 .01) .②The difference SNR among different PTA groups were significant (P<0 .01) ,and when the influence of MMSE scores was controlled the difference was still significant .There was a significant linear correlation between PTA and SNR (r=0 .326 ,P<0 .01) .MMSE scores decreased with the increasing of PTA ,and there was also a linear correlation between PTA and MMSE scores (-0 .187 ,P<0 .05) .③Adaptive SNR and MMSE scores were correlated (r= -0 .201 ,P<0 .01) ,but there was no significant linear relationship .Besides ,SNR of the cog_nitive function impairment group (<27 points) was significantly greater than that of the normal cognitive function group (27~30 points) (P<0 .01) .However ,among the three PTA groups ,the differences were not statistically significant .ConcIusion The hearing in noise test reflects the ability of speech recognition in noise enviroment in ol_der adults ,and can be used to assess the hearing functions in elder .Speech recognition in noise ,peripheral hearing loss and cognitive functions are relevant .Cognitive functions have an influence on the evaluation of the hearing func_tion for older adults .
9.HHIE-S Scale for Hearing Screening in the Elderly Population
Xianting HU ; Zhiwu HUANG ; Jianyong CHEN ; Ling MEI ; Yun LI ; Yan REN ; Zhijian TANG ; Min SHEN ; Qin ZHANG ; Hao WU
Journal of Audiology and Speech Pathology 2014;(3):230-234
Objective To investigate the reliability and practicality of Chinese version the hearing handicap in-ventory for the elderly -screening version(HHIE -S) scale for elderly hearing screening in China .Methods We compared the HHIE -S scores with results of pure -tone audiometry test .A total of 840 Chinese individuals older than 60 years filled in the scale all by themselves first ,then refilled it according to explanations of each question translated by researchers .All participants finished pure -tone audiometry test with frequencies of 0 .25 ,0 .5 ,1 .0 , 2 .0 ,and 4 .0 kHz .Sensitivity ,specificity ,positive predictive value (PPV ) ,false-negative rate and false -positive rate were compared with HHIE -S cut -off points of >8 or >10 and pure-tone averages (PTAs) greater than 25 ,40 and 60 dB HL .Analysis the area under the curve (AUC) values of receiver operating characteristic (ROC) curves for each three PTA levels .Results Cronbach's α of the scale was 0 .85 .HHIE -S scale pre -explanatory score was 7 .6 ± 10 .6 ,and post-explanatory score was 10 .8 ± 16 .7(P<0 .01) .HHIE -S> 8 points higher than H HIE-S> 10 points in the sensitivity ,positive predictive value increase ,specificity ,false-positive rate ,false negative rate decreased ,paired samples t - test sensitivity and false -negative rate (P< 0 .05) .After the guide meanwhile HHIE-S cut-off points of > 8 when grouping with PTA> 25 dB HL ,PTA> 40 dB HL and PTA>60 dB HL ,conducted ROC curve analysis ,the AUC value was 0 .70 ± 0 .03 ,0 .84 ± 0 .01 and 0 .88 ± 0 .02 .Conclusion The score of HHIE-S scale>8 points can be used when PTA> 40 dB HL as goal for hearing loss of old people , it can be more accurate after explanations ,so HHIE-S scale can be used as a preliminary hearing screening method for Chinese elderly people .
10.Interaction of folate deficiency and aberrant profile of DNA methyltransferase 1 in the progression of cervix carcinogenesis.
Jin-tao WANG ; Ling DING ; Jun-xia HAO ; Wei-min ZHAO ; Qin ZHOU ; Min HAO
Chinese Journal of Preventive Medicine 2012;46(12):1088-1094
OBJECTIVETo explore the interaction of folate deficiency and aberration of DNA methyltransferase 1 (DNMT1) in the progression of cervix carcinogenesis.
METHODSAll clinical samples were collected from 80 patients with cervix squamous cell carcinoma (SCC), 105 patients with cervical intraepithelial neoplasm (CINI, n = 52; CINII/III, n = 53) and 53 patients with cervix inflammation (CI). The participants were diagnosed by histology at Shanxi Province Tumor Hospital and Second Hospital of Shanxi Medical University during the period of September 2009 to May 2010. Meanwhile, cervical cancer cell lines Caski and C33A were treated with different concentration of folate. Radioimmunoassay (RIA), Western blotting and real-time PCR were used to detect the levels of serum folate, the expression of DNMT1 protein and mRNA, respectively. The data were analyzed by Student t test, ANOVA, chi-square test and Spearman correlation using SPSS statistical package. The correlation strength between factors and cervical canceration was calculated by OR and 95%CI value. Interaction effect was evaluated by the application of additive effect model.
RESULTSThe levels of serum folate (median inter-quartile range) were (2.66 ± 1.82), (2.83 ± 2.23), (3.17 ± 1.91) and (3.21 ± 1.74) ng/ml, the levels of DNMT1 protein (x(-) ± s) were 2.28 ± 0.55, 1.84 ± 0.37, 1.33 ± 0.38 and 0.92 ± 0.29, the Ct-ratio (Ct value of DNMT1/Ct value of β-actin) of DNMT1 mRNA (x(-) ± s) were 1.26 ± 0.13, 1.27 ± 0.12, 1.27 ± 0.12 and 1.33 ± 0.11 in the group of SCC, CINII/III, CINIand CI, respectively. The results showed that the serum folate levels were descended, and the expression levels of DNMT1 protein (χ(2)(tend) = 50.80, P < 0.05) and mRNA (χ(2)(tend) = 17.63, P < 0.05) were increased steadily with the severity of the cervix lesions. Moreover, our results revealed that there was an additive interaction between folate deficiency and high-expression of DNMT1 protein related to the risk of CIN and SCC. And it showed that the relative excess risk of interaction (RERI), attributable proportion of interaction (API) and synergy index(S) was 0.27, 0.14 and 1.40 in CINI group, 0.47, 0.19, 1.46 in CINII/III group, 1.60, 0.31, 1.61 in SCC group, respectively. It was found that folate was able to reduce the proliferation of Caski and C33A cells (r values were 0.954 and 0.969, all P values < 0.05), with 11.4% and 13.6% of growth inhibition at the concentration of 10 µg/ml, 64.8% and 49.4% at 1000 µg/ml in Caski and C33A cells, respectively. The result showed there was an inverse correlation between the levels of folate and DNMT1 protein (r values were -0.859 and -0.914, all P values < 0.05), with 1.96 and 1.92 of expression levels at the concentration of 10 µg/ml, and 1.60 and 1.38 at 1000 µg/ml in Caski and C33A cells, respectively. At folate concentration of 1000 µg/ml, the expression of DNMT1 protein or mRNA was higher in Caski cell than in C33A cell (t values were -4.22 and 3.50, all P values < 0.05).
CONCLUSIONOur finding indicated that the low levels of serum folate and high-expression of DNMT1 protein or mRNA seemed to be associated with high risk of cervical cancer and cervix precancerous lesion. Sufficient folate is able to effectively inhibit the growth of cervical cancer cells in vitro, and would counteract transcriptional and posttranscriptional aberration of DNMT1. It suggested that there might be a synergistic action between folate deficiency and aberration of DNMT1 in the progression of cervix carcinogenesis.
Adult ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; metabolism ; pathology ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; metabolism ; Female ; Folic Acid ; blood ; Folic Acid Deficiency ; metabolism ; Humans ; Middle Aged ; RNA, Messenger ; genetics ; Uterine Cervical Neoplasms ; metabolism ; pathology