1.Sequence analysis of TraE in the novel conjugative plasmid pO157_Sal detected in Escherichia coli O157∶H7
Chinese Journal of Infectious Diseases 2014;(7):13-16
Objective To analyze the sequence of the novel conjugative plasmid pO157_Sal detected in outbreak isolates of Escherichia coli O157∶H7 .Methods The traE genes of the outbreak isolates in China were amplified by polymerase chain reaction (PCR) and the products were sequenced .The TraE sequences of Escherichia coli O157 ∶ H7 strains from other sources were retrieved from GenBank . Phylogenetic tree based on the TraE sequences was constructed by Neibhor-joining analysis .The whole plasmid sequences of pO157_Sal and pEC4115 were compared .Results The sequences of traE gene were identical among the Chinese isolates . There were homologous sequences of TraE in Escherichia coli O157∶H7 isolates from different sources .Twenty-one out of the 52 pO157_Sal genes were homologous to genes of pEC4115 with amino acid level identity ranging from 28% to 51% .Conclusions Although similar TraE sequences and similar plasmid are found in Escherichia coli O157∶H7 isolates from different sources ,pO157_Sal is only observed in Chinese outbreak isolates .The TraE sequences are conservative among the outbreak isolates ,indicating they are from the same specific source .
2.Development of a novel automatic water system for ventilator humidifier
Yinchun LU ; Hongqing ZHAO ; Jingying GAO
International Journal of Biomedical Engineering 2014;37(6):368-370
Objective To develop a novel automatic water system for ventilator humidifier.Methods By hanging the infusion apparatus high,sustained water was filled by driving force of gravity.Liquid level in the thong of the infusion apparatus was detected by the sensor,and the closing and opening of the closing clip was controlled by solenoid valve.Results The closing clip opened when the liquid level inside the ventilator humidifier fell below the lowest setting value and purified water in the infusion bottle automatically flowed to the humidifier.When the liquid level reached the highest setting level,the closing clip was automatically closed.Conclusions The developed automatic water system for ventilator humidifier is effective,convenient,inexpensive,and realized a sustained,relatively steady and a small amount of water process.Meanwhile,humidifier water is stable,heating is uniform,and gas temperature is relatively constant,which can be applied in clinical use.
3.Clinical incremental values of extraosseous findings on CT during bone SPECT/CT imaging
Xiangyun ZHU ; Hongqing ZHAO ; Yanjun ZHAO ; Ping TANG ; Jianming NI
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(7):400-403
Objective To evaluate the prevalence of extraosseous findings on integrated CT images of routine SPECT/CT bone imaging and its clinical incremental values. Methods A total of 843 patients (470 males, 373 females, age range: 26-92 years) who underwent SPECT/CT bone imaging during May 2013 to December 2015 were enrolled in this retrospective study. A modified C-RADS was used to classify the extraosseous findings to E1, E2, E3 and E4. χ2 test was used for data analysis. Results The CT images in 78.6%(663/843) of patients were normal or with no additional clinical significance (E1 and E2), and those in 21.4%(180/843) of patients might need further assessment (E3 and E4). The rate of E4 extraosseous findings in patients with malignancy was higher than that in patients without malignancy: 9.5%(59/622) vs 5.0%(11/221); χ2=4.352, P<0.05. There was no significant difference of the rate between genders: 8.5%(40/470) in males vs 8.0%(30/373) in females; χ2=0.510, P>0.05. With age increasing, the prevalence of E4 finding increased and the rate was the highest in the patients over 80 years old (125%, 16/128). Seventy patients had E4 findings and chest masses and nodules were the most common, followed by the abdominal or pelvis lymph node enlargements. Conclusions Potentially important extraosseous findings are common on SPECT/CT. Systematic reviewing CT images and communicating the important unexpected findings to clinical physicians could enhance its clinical incremental values.
4.Cloning and bioinformatics analysis of SBACT5 gene in Echinococcus multilocularis from Southern Qinghai Plateau
Shunwei HE ; Xiaoyan LI ; Hongqing LI ; Ruixue ZHAO ; Xiaoxing WEI
Journal of Medical Postgraduates 2017;30(8):818-823
Objective Sodium-bile acid cotransporter plays an important role in the development of Echinococcus.The present study aimed to clone sodium bile acid cotransporter gene in Echinococcus multilocularis (EmSBACT5) and to analyze the bioinformatics of its coding protein.Methods EmSBACT5 gene was amplified by reverse transcription RCR (RT-PCR) technology and its nucleotide sequence was sequenced.Bioinformatics softwares were used to predict and analyze the physical and chemical properties, hydrophilicity/hydrophobicity, transmembrane domain, post-translational modification sites, structural domain, secondary structure, tertiary structure, subcellular localization and biological functions of the coding protein.Results The complete open reading frame was amplified with 654 bp in length, encoding 217 amino acids.The homology of the nucleic acid sequence and amino acid sequence of EmSBACT5 gene were 98% and 96% with the published SBACT5 in Echinococcus granulosus (EgSBACT5) respectively.Protein analysis results showed that the molecular formula of EmSBACT5 protein was C1141H1797N273O284S11.Its relative molecular mass was 24240 and isoelectric point was 8.99.There were 9 post-translational modification sites and 4 typical domains.Alpha helical, β-sheet, β-turn and random coil accounted for 29.95%, 31.80%, 7.83% and 30.41%, respectively.This protein was a hydrophobic membrane protein and was mainly located in the cytoplasmic membrane, and it might play a role in the processes of material transport and signal transmission.Conclusion The EmSBACT5 gene was cloned successfully and the informatics characteristics of its coding protein were obtained, which provides basic information for prevention and control of echinococcosis.
5.Distribution of intestinal bacteria genera between normal weight and overweight/obesity school-age children in Yili region of Xinjiang
Li LIU ; Peiru XU ; Min LI ; Hongqing WANG ; Yuli ZHAO ; Wenchao LI
Chinese Journal of Applied Clinical Pediatrics 2014;29(7):502-505
Objective To investigate intestinal bacteria genera distribution between normal weight and overweight/obesity school-age children in Yili region of Xinjiang.Methods Selecting eligible 150 fecal samples from school-age children (aged from 7 to 13 years old),all samples were divided into normal weight group and overweight/obesity group according to the body mass index (BMI),and each group contained 75 samples.Fecal samples were collected and DNA was extracted,then 6 types of intestinal bacteria genera were detected by the quantitative real-time polymerase chain reaction.Results The distribution of age,gender,ethnicity and hip circumference between 2 groups had no statistically significance (all P > 0.05),except that of BMI and waist circumference (t = 20.740,8.533,all P < 0.01).The concentration of Lactobacillus,Clostridium and Enterococcus were significantly higher in the obese/overweight children (t =9.735,9.681,26.070,all P < 0.01),whereas no significant differences were found in the concentration of Escherichia,Bifidobacterium and Bacteroides between 2 groups.According to the stratified analysis of ethnicity,gender and age,there was no significant difference among Han,Kazakh and Uyghur children.While stratified by gender,Bifidobactcrium was significantly lower in girls than that of boys (t =4.931,P < 0.05).The distribution of the 6 types of intestinal bacteria genera was no statistically significant among different ethnicity,gender and age groups.Conclusions The intestinal bacteria genera number distribution in different ethnicity,gender and age groups makes no sense.The 16SrRNA type number of the Lactobacillus,Clostridium and Enterococcus may be associated with childhood obesity.
6.Enhancement of radiation sensitivity by erlotinib and celecoxib in A549 human lung cancer cell line
Jian SUN ; Ningbo LIU ; Hongqing ZHUANG ; Lujun ZHAO ; Zhiyong YUAN ; Ping WANG
Chinese Journal of Radiological Medicine and Protection 2012;32(2):186-190
Objective To investigate the role of epidermal growth factor receptor and cyclooxygenase-2 pathways in the erlotinib and celecoxib enhanced radiation sensitivity in A549 human lung cancer cell line. Methods IC20 of erlotinib and celecoxibon in A549 human lung cancer cells was measured by MTT assay,Clonogenic assays were used to evaluate the antitumor effects of the drugs and Xirradiation.Flow cytometry was used to assess the apoptosis and cell cycle alteration,and Western blot was used for the detection of Akt and phospho-Akt.Results Both erlotinib and celecoxib could inhibit the proliferation of A549 cells in vitro in a dose-dependent manner and their values of IC20 were (5.15 ± 0.14)and (40.32 ± 1.26) μmol/L,respectively. For radiation survival,the values of Dq,Do,SF2 of the combination of two drugs were lower than those of either drug ( t =6.62,P < 0.05).The SER of celecoxib,erlotinib and their combination were 1.299,1.503 and 2.217,respectively.Flow cytometry assay showed that both celecoxib and erlotinib could enhance radiation-induced G0/G1 arrest,reduce the cell numbmer in S phase,and enhance radiation-induced apoptosis,especially for the combination of drugs.Western blot assay showed that the expressions of Akt protein were similar in all groups.However,pAkt expression was suppresssed by erlotinib and celecoxib,but promoted by radiation.pAkt had the lowest expression in the radiated cells with the treatment of two drugs ( t =4.89,P < 0.05 ).Conclusions The edotinib and/or celecoxib could enhance radiosensitivity probably by increasing cell apoptosis and reducing the number of Sphase cells with low radiosensitivity.
7.Radiosensitization of recombinant human endostatin in human lung squamous cancer cells in vitro
Zhenyu YOU ; Junjie WANG ; Yong ZHAO ; Hongqing ZHUANG ; Feng LIU ; Yingdong ZHANG
Chinese Journal of Radiation Oncology 2009;18(4):326-329
Objective To investigate the radiosensitising effect of recombinant human endostatin (endostar) on human lung squamous cancer cell line H-520 in vitro and its mechanism. Methods H-520 cells in exponential growing phase were treated with endostar alone, irradiation alone, or endostar plus irra-diation. Colony-forming assay was used to investigate the cytotoxicity and radiosensitising effects of endostar. Cell survival fractions of all groups were calculated and cell survival curves were fitted by single-hit multi-tar-get model. Cell apoptosis, cell cycle distribution and activated Caspase expression level were investigated by flow cytometry. Results The D0, Dq, D10 and SF2 values of combined treatment group were much lower than those of irradiation alone group. The sensitization enhancement ratio (SER) was 1.50 (ratio of D0 values). Endnstar induced H-520 cell apoptosis in a dose dependant manner. After administration of endostar, H-520 cell proliferation was inhibited, and cell apoptosis rate and apoptotic bodies were increased. After irradiation of 0 Gy, 2 Gy, 4 Gy and 8 Gy, the apoptosis rate of H-520 cells was 4.27% ±0.29%, 14.3% ±1.15%, 28.49% ± 1.58% and 54.79% ± 1.89% in the radiotherapy alone group, and 22.38% ± 1.61%, 35.01% ±1.16%, 46.83%±2.06% and 64.08%±4.28% in the combined treatment group, respective-ly. The difference between the two groups was significant (t = 19.17, 17.79, 25.64 and 3.44,all P < 0.05 ). Flow cytometric analysis showed that cell cycle distribution changed and G0 + G1 phase arrest oc-curred after endostar treatment, while irradiation induced G2 + M arrest. The expression level of activated Caspase in combination group (62.7% ±1.9% ) was higher compared to the control group ( 12.1%± 0. 1% ) , endostar alone group ( 54.6% ±1.0% ) and irradiation alone group ( 34.1%±1.2% ) ( t = 46.69, 6.55 and 22.54 ; all P < 0.05 ). Conclusion Endostar can enhance the radiosensitivity of H-520 ceils by inhibiting cell proliferation, promoting cell apoptosis and facilitating cell cycle redistribution.
8.Distribution of six genes of essential hypertension In Yunnan Han healthy population
Xiaoli ZHAO ; Dachun HU ; Jianchun SHAO ; Jing QIAN ; Hongqing ZHANG ; Jie JIANG
Clinical Medicine of China 2009;25(2):135-137
Objective To explore the distribution of RAS,AGT,ACE,eNOS,ET-2,ANP and NPRC of es-sential hypertension in Yunnan Han healthy population.Methods Gene chip technology was used to detect the pol-ymorphism of AGT M235T (MM, MT,TT), ACE I/D (II, ID, DD ), eNOS Glu298Asp (EE, ED, DD), ET-2 A985G (AA,AG,GG) ,ANP T2238C(TT,TC,CC) and NPRC A-55C(AA,AC,CC) in 97 health subjects.Results The MM,MT and TT genotype frequency of AGT M235T was 0.052,0.381 and 0.567 ,alle frequency of M and T was 0.242 and 0.758 in 97 healthy subjects of Yunnan population;The II, ID and DD frequency of ACE I/D was 0.340, 0.598 and 0.062, alle frequency of I and D was 0.680 and 0.320 in 97 healthy subjects of Yunnan population ;EE, ED and DD frequency of eNOS Glu 298 Asp was 0.845,0.144 and 0.011 ,alle frequency of E and D was 0.918 and 0.082 in 97 healthy subjects of Yannan population;AA,AG,GG frequency of ET-2 A985G was 0.020,0.258 and 0.722, alle frequency of A and G was 0.149 and 0.851 in 97 healthy subjects of Yunnan population;TT and TC fre-quency of ANP T2238C was 0.959 and 0.041 ,and CC was not detected;alle frequency of T and C was 0.979 and 0.021 in 97 healthy subjects of Yunnan population;AC and CC frequency of NPRC A-55C was 0.763 and 0.237, and no AA was detected,alle frequency of A and C was 0.381 and 0.619 in 97 healthy subjects of Yunnan popula-tion.Conclusion The polymorpbism of ACT M235T,ACE I/D,eNOS Glu298Asp,ET-2 A985G,ANP T2238C and NPRC A-55C is locally distributed in Yunnan Han healthy population.
9.Gefitinib at different administration time in radiosensitizing lung adenocarcinoma cells in vitro
Peiyan QIN ; Hongqing ZHUANG ; Zhiyong YUAN ; Jun WANG ; Lujun ZHAO ; Ping WANG
Chinese Journal of Radiation Oncology 2010;19(2):150-152
Objective To investigate the radiosensitizing effects of gefitinib at different administration time. Methods Gefitinib was administered to A549 lung cancer cells in three different ways (method 1, 24 h before irradiation ;method 2, upon irradiation and method 3, 24 h after irradiation). Cell-surviving rates were evaluated by the colony-forming assays. Cell apoptnsis and cell-cycle distributions were detected by the flow cytometry (FCM). Protein expression of p21, Cdc25c, Bcl-2, Bax, Rad51 and phosphorylated DNA - PKcs (phnspho - DNA - PK) were measured with the Western blot analysis. Results The sensitizing effect ratio (ratio of D_0 value) was 2.23, 1.51 and 1.30 with method 1, 2 and 3, respectively. A higher apoptosis rate and more G_2/M phase arrest were observed with method 1 when compare with method 2 or 3. With the similar tendency, the protein level of p21, Cdc25c, Bcl-2, Bax, RadSl and phospho-DNA-PK changed distinctly. Conclusions Radiosensitizing effects are obtained in all three methods, with gefitinib delivered before irradiation being the best.
10.Changes of content of monoamine neurotransmitters and expression of neurotrophic factors in brain regions of rat models of anxious depression
Hongqing ZHAO ; Yuanshan HAN ; Zhuo LIU ; Qing DU ; Qin YANG ; Pan MENG ; Hui YANG ; Yuhong WANG
Acta Laboratorium Animalis Scientia Sinica 2017;25(4):373-379
Objective To study the content of monoamine neurotransmitters and neurotrophic factor in the hippocampus, amygdala and prefrontal cortex in anxious depression rats, and explore the possible pathogenesis.Methods 60 SD rats were randomly divided into normal group, vehicle group, anxiety group, depression group, and anxious depression group, 12 rats in each group.Chronic restraint stress combined with corticosterone injection was used to establish anxiety and depression model, the modeling time was 21 d.After modeling, elevated plus maze test, open field test, and forced swimming test were used to evaluate the anxiety and depression-like behavior, HPLC-ECD was used to detect the content of 5-HT, NE, and DA in the hippocampus, amygdala, and prefrontal cortex of rats.Western-blotting was used to detect the expression of BDNF and NT-3 in rats.Results Rats in anxious depression model group were comparable to the anxiety group in time and frequency entering open arm time, and number of locomotor activity in open field, and it had a significant difference when compared with the control and depression groups (P<0.01 or P<0.05).Immobile time in anxious depression model rats was increased significantly when compared with the control and anxiety groups (P<0.01).Meanwhile, compared with the control group, 5-HT in hippocampus and 5-HT, NE in amygdala or prefrontal cortex were significantly decreased in the depressive rats with anxiety (P<0.01 or P<0.05).Moreover, the content of BDNF and NT-3 was significantly decreased in each brain regions compared with the control group (P<0.01 or P<0.05), and BDNF levels were obviously decreased compared with the anxiety group (P<0.05).Conclusions Rats of anxious depression have significant anxiety and depression-like behaviors.Its mechanism may be associated with the down-regulation of monoamine neurotransmitters and neurotrophic factors BDNF and NT-3 in hippocampus, amygdala, and prefrontal cortex region.