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 .

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.

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 (125%, 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.

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.

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.

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.

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.

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.

DCD WeChat ethical review had its necessity and superiority but problems and confusion as well. For instance, WeChat review time was urgent and limited;WeChat review discussed insufficiently even hard to car-ry out;the methods of WeChat review were limited;the authenticity of audit opinion was difficult to grasp;the re-view data privacy existed hidden danger;the review quality was difficult to guarantee;WeChat conference content file archive was difficult, and so on. As to this, the author put forward the following countermeasures:giving war-ranty on the ethical review time as far as possible;initiating the committee review by the referee and uploading the review information to WeChat group, then ethical reviewers expressing their opinions in real names to ensure data confidentiality and privacy protection; strengthening the ethics committee education for organ transplantation; im-proving the quality of the review committee members and moral cultivation;archiving the WeChat conference infor-mation.

Objective To compare the biological characteristics of several different anxiety rat models established by different methods of stress at different time points and provide experimental basis for the most appropriate modeling methods .Methods 60 rats were randomly divided into normal , empty bottle stress , chronic emotional stress ( CES ) group, restraint stress for 3h, 6h, and modeling respectively .In the experimental 7 d, 14 d, 21 d, elevated plus maze and fear condition system was used to test anxiety-like behavior in rats , open field test to study anxiety or depression-like behavior , forced swimming test was used to detect depression-like behavior in rats , and using the Elisa test kit to detect the contents of 5-HT, DA in the hippocampus in rats .Results Anxiety-like behavioral test results showed that rats in empty bottle stress, CES, 6 h restraint stress group started to have anxiety-like behavior since 14 d, then anxiety-like behavior was becoming increasingly apparent .Forced swimming test results showed that immobility time in 6 h restraint rats was significantly increased in the first 7 d(P <0.05).Meanwhile, compared with control group, hippocampal 5-HT, DA contents in empty bottle stress and CES rats increased significantly since 14 d.Conclusions Among several stress methods established anxiety model , anxiety-like behavior in 3 h restraint stress was not obvious; 6 h restraint stress exhibited a depression-like behavior in the forced swimming test might be due to prolonged stress .Empty bottle stress and CES can successfully establish the anxiety rat model , and the anxiety behavior of the rats have some differences . Corresponding model methods can be selected according to different experimental purposes .