Objective To observe the changes of TNF-α and NF-κB after different doses of nalmefene hydrochloride (NAL) therapy for traumatic brain injury (TBI) in an effort to identify the effect of NAL on TBI-induced inflammatory response and the possible mechanism.Methods A model of TBI in the rat was produced using the improved Feeney' s free-fall impact method.The animals were randomly divided into sham group,TBI group,TBI + large dose of NAL (ip,0.2 mg/kg) group (TBI + NAL1group),TBI + medial dose of NAL (ip,0.14 mg/kg) group (TBI + NAL2 group),TBI + small dose of NAL (ip,0.07 mg/kg) group (TBI + NAL3 group).Form of brain tissues in each group was observed and mRNA levels of TNF-α and NF-κB were measured by real-time quantitative PCR assay.Results HE staining revealed severe injury and inflammatory infiltration of brain parenchyma in TBI group ;on the contrary,the situation ameliorated in TBI + NAL1 group,TBI + NAL2 group and TBI + NAL3group,with especially obvious improvement in TBI + NAL2 group.In PCR assay,significant expression of NF-κB and TNF-α was observed at post-TBI days 1,3,5 and 7 (P ＜ 0.05),followed by great reverse after NAL therapy (P ＜ 0.05),particularly in TBI + NAL2 group.Conclusions NAL can reduce the inflammation response to TBI and promote post-injury recovery.Moreover,there exists a NAL concentration window.

BACKGROUND:C-reactive protein has been shown to rapidly increase during the occurrence of inflammation and tissue injury, and can indicate the degree of inflammatory reaction.
OBJECTIVE:To analyze the correlation between survival time and C-reactive protein in rabbits after transplantation of polytetrafluoroethene artificial trachea with support ring.
METHODS:The cervical trachea of rabbits was replaced by polytetrafluoroethene artificial trachea with support ring. Survival time of the rabbit, and the changes in serum C-reactive protein at 1-7 days after transplantation were observed. Linear regression was used to assess the univariate association between serum C-reactive protein and survival time.
RESULTS AND CONCLUSION:The linear correlation was observed between changes of serum C-reactive protein and survival time in rabbits with artificial trachea replacement operation. C-reactive protein levels in rabbits with<13 days of survival time were increased and positively associated with the number of days after transplantation. However, C-reactive protein levels in rabbits with>13 days were decreased and negatively associated with the number of days after transplantation. In rabbits with positive correlation and negative correlation, the median survival time and 95%confidence interval (CI) were respectively 10 days (95%CI 8.614-11.386 days) and 27 days (95%CI 23.970-30.030 days). The survival rate in negative correlation group was significantly higher than positive correlation group (x2=29.364, P<0.01). Results suggested that the prolonged survival time of rabbits after artificial trachea replacement operation was related to the decreased concentration of serum C-reactive protein.

Two kinds of β2-agonistresidues in sheep plasma and urine were disposed by enzymolysis and organic solvent extraction pretreatment methods, and UPLC-MS/MS was used for the qualitative and quantitative analysis. Detection results were compared to study the influences of two pretreatment methods. The experimental results showed that more than 95% of Ractopamine and 40% of Salbutamol exist in the conjugated form in sheep plasma. The detection results of 2 kinds of β2-agonist residues were significantly enhanced when adding β-glucuronidase/aryl sulfatase. The experimental repeatability is very poor ( RSD>40%) when the enzymolysis was not carried out. There were 57% of Ractopamine and less than 1% of Salbutamol exists in the conjugated form in sheep urine. Enzymolysis pretreatment method was useful for the Ractopamine residues determination in urine, and Enzymolysis pretreatment method was useless for Salbutamol determination in urine. Matrix effect of plasma was less than the effects of urine. The influence of organic solvent extraction pretreatment method on the detection results was unremarkable, and there was the possibility that organic solvent extraction could lead partial loss of target compound in extraction process. However, it did not influence the detection results by using internal standard calibration.

To express Pleurocidin in Escherichia coli and to enhance the secretory efficiency of the fusion protein, the gene encoding Pleurocidin was ligated with Cherry DNA sequence via blunt-end ligation. Then this fusion gene was cloned into pET22b (+) vector and the recombinant plasmid was transformed into E. coli BL21 (DE3). Lactose was used to induce expression of fusion protein. The recombinant plasmid pET22b (+) -CP was successfully constructed and high-level expression of fusion protein was induced with lactose. Statistics showed that addition of glycine after 16 h of induction significantly enhanced the secretory efficiency of the fusion protein. After hydrolysis of the fusion protein by diluted hydrochloric acid and some further purification steps, r-Pleurocidin was obtained with antibacterial activity against E. coli DH5α and Bacillus subtilis BS168. In conclusion, the fusion protein was expressed in E. coli and biologically active r-Pleurocidin was obtained after hydrochloric acid cleavage and purification.
Animals ; Cloning, Molecular ; Escherichia coli ; metabolism ; Fish Proteins ; biosynthesis ; Flounder ; Recombinant Fusion Proteins ; biosynthesis

Objective To establish a direct reverse transcription real-time fluorescence quantitative polymerase chain reaction ( RT-qPCR-D ) method for detecting serum circulating B cell-specific moloney murine leukemia virus integration site-1 (Bmi-1) mRNA, and analyze the levels of serum circulating Bmi-1 mRNA in colorectal cancer patients by using of this method for exploring its diagnosis value in colorectal cancer.Methods Methodology establishment.RNA was extracted from colorectal cancer HT 29 cell line, and detection standard curves of Bmi-1, ubiquitin C ( UBC), glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) mRNAs were established , then the amplification efficiencies were calculated.Bmi-1 mRNA level was directly detected in serum and preparation buffer mixture , then the specificity of assay was evaluated by melting curve, and detection limit was observed through diluted serum samples.The serum circulating Bmi-1 mRNA levels were detected by ELISA in 158 cases with colorectal cancer , of which there were 26 cases of tumor node metastasis ( TNM)Ⅰstage, 53 cases of TNMⅡ, 47 cases of TNMⅢ, 32 cases of TNMⅣand 53 cases of controls with normal colonoscopy collected from January 2008 to January 2009 in Qilu Hospital of Shandong University.Comparisons of groups were determined by applying Mann-Whitney U test or Kruskal-Wallis test, and receiver operating characteristic ( ROC) curves were established to illustrate the diagnostic performance.Results The log values of Bmi-1, UBC and GAPDH showed good linear correlations with quantification cycle (Cq) values(R2 =0.990, 0.990, 0.991, all P <0.001), and the amplification efficiencies were 0.875, 0.917 and 0.935, respectively.Using the established RT-qPCR-D method, the peak of melting curve of Bmi-1, UBC and GAPDH mRNAs were single, the detection limit was up to 1.25μl.The levels of serum circulating Bmi-1 mRNA detected by RT-qPCR-D were 0.138 ( 0.078-0.228 ) in colorectal cancer stage Ⅰ patients, 0.163(0.067 -0.287) instage Ⅱ patients, 0.217(0.072-0.267) instage Ⅲpatients, 0.273(0.139 -0.419) in stage Ⅳ patients and 0.021(0.008 -0.029) in health controls, a significant difference was found among groups ( H =89.5, P <0.001 ).The levels of serum circulating Bmi-1 mRNA in each stage colorectal cancer were all significantly higher than that in control group(U=58.0, 287, 246, 72.5,all P<0.001).The levels in Ⅳstage patients were significantly higher than those in other stages patients (U=247, 590, 540,P=0.008, 0.020, 0.035), while no significant differences among Ⅰstage,Ⅱstage and Ⅲstage patients(U=633, 514, 1170,all P>0.05).ROC curve analysis showed area under the ROC curve ( AUC) for serum circulating Bmi-1 mRNA was 0.921(95%CI=0.876-0.953), which was significantly superior to the AUC of CEA (0.745, 95%CI=0.680-0.802, Z=4.697, P<0.001 ).When cutoff value was 0.034, the diagnostic sensitivity and specificity was 89.2%(141/158) and 90.6%(48/53), while 41.8%(66/158) and 73.6%(39/53) using CEA.The AUC for combination of circulating Bmi-1 mRNA and CEA was 0.933(95%CI=0.890-0.963 ) , which was no statistical significance when compared with the AUC of circulating Bmi-1 mRNA(Z=4.697, P>0.05).Conclusions The study establishes a higher sensitive, specific for detecting serum circulating Bmi-1 mRNA. Based on this method , serum circulating Bmi-1 mRNA is found to be increased in colorectal cancer , and is superior to traditional tumor marker CEA in diagnosis of colorectal cancer, which may become a potential detection index for early detection of colorectal cancer.

Enzymes can degrade the anti-nutrient factors in feedstuff, increase nutrient digestibility, and reduce pollution to environment, and have been widely supplemented in animal feedstuff. However, the use of enzymes is limited because of their undesirable properties, such as thermoliability and susceptibility against protease digestions. And its commercialization is also limited by low production efficiency and high cost. Therefore, the focuses for future enzyme development will be: (1) to obtain novel enzymes with better properties by high-throughput screening of enzyme encoding genes, especially those from extreme and special environments; (2) to improve enzyme properties using directed mutagenesis and protein engineering methods; (3) to achieve high-level fermentation of enzymes by heterogonous expression and optimization of codons, vectors and fermentation conditions; (4) to determine the effect of enzymes to animals and utilize enzymes efficiently.
6-Phytase ; genetics ; metabolism ; pharmacology ; Animal Feed ; Animals ; Dietary Supplements ; Lipase ; genetics ; metabolism ; pharmacology ; Peptide Hydrolases ; genetics ; metabolism ; pharmacology ; Protein Engineering

OBJECTIVETo correlate morphological features with mutations of epidermal growth factor receptor (EGFR) in lung adenocarcinomas.

METHODSAccording to 2011 International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society International Multidisciplinary Lung Adenocarcinoma Classification, a total of 72 surgically resected lung adenocarcinomas were collected and classified into different histological subtypes and different cell types (hobnail, columnar and polygonal). EGFR gene mutation was detected with the amplification refractory mutation method provided by the EGFR mutation test kit. The correlation between these subtypes and EGFR mutations were evaluated.

RESULTSMutations of EGFR were detected in 48.6% (35/72) of lung adenocarcinomas; 19del and L858R were major mutational types (88.6%, 31/35). EGFR mutations were associated with female gender, non-smoking status, and well to moderately differentiated tumor histology. EGFR mutation types were not associated with age, smoking index, lymph node metastasis, stage, status of whether have or not have inclusion bodies or psammoma bodies and mitotic level. Correlations were observed between acinar and papillary adenocarcinoma subtypes and EGFR mutations according to the new classification. EGFR mutation was rare in the subtype of solid adenocarcinoma with mucin production and almost never observed in special subtypes (mainly mucinous and colloid adenocarcinoma). In addition, EGFR mutation was associated with the hobnail cell type.

CONCLUSIONLung adenocarcinomas of predominate acinar and papillary histological subtypes with hobnail cell morphology are good predictors for EGFR mutations.

Adenocarcinoma ; genetics ; pathology ; Adenocarcinoma, Mucinous ; genetics ; pathology ; Adenocarcinoma, Papillary ; genetics ; pathology ; Female ; Genes, erbB-1 ; Humans ; Lung Neoplasms ; genetics ; pathology ; Lymphatic Metastasis ; Male ; Mutation ; Receptor, Epidermal Growth Factor ; genetics ; Sex Factors ; Smoking

Objective? To investigate the current development of hospital nursing information systems from the perspective of nurses. Methods? According to the characteristics of nursing information system, self-made questionnaires were published online from 3rd to 10th May, 2018 in Weixin Subscription Number of Zhejiang Zhongwei Nursing Information Management Research Institute and all kinds of Weixin groups related to nursing management/information. A total of 1 966 nurses volunteered to participate in the survey and 1 364 valid questionnaires were collected. Results? A total of 417 hospitals were covered in the survey, including 281 tertiary hospitals and 130 secondary hospitals. 82.75% of the tertiary hospitals and 65.65% of the secondary hospitals had nursing information system. The nursing information systems were mainly based on the clinical nursing information system and the nursing management information system. The proportion of clinical nursing information system "meeting clinical nursing needs very well" was 5.21%. The proportion of "satisfying nursing management needs very well" in nursing management information system was 1.96%. There are some problems in nursing information system, such as "each subsystem can not be docked" "data can not be extracted","data need to be re-entered" and so on. Conclusions? The construction of hospital nursing information system in China is relatively backward, and nursing information system can not meet the needs of clinical and management. In order to build intelligent nursing information system, it is urgent to carry out research on nursing standard terminology.

Objective To correlate morphological features with mutations of epidermal growth factor receptor ( EGFR) in lung adenocarcinomas.Methods According to 2011 International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society International Multidisciplinary Lung Adenocarcinoma Classification, a total of 72 surgically resected lung adenocarcinomas were collected and classified into different histological subtypes and different cell types ( hobnail, columnar and polygonal ) . EGFR gene mutation was detected with the amplification refractory mutation method provided by the EGFR mutation test kit.The correlation between these subtypes and EGFR mutations were evaluated.Results Mutations of EGFR were detected in 48.6%(35/72) of lung adenocarcinomas;19del and L858R were major mutational types (88.6%,31/35).EGFR mutations were associated with female gender, non-smoking status, and well to moderately differentiated tumor histology.EGFR mutation types were not associated with age, smoking index, lymph node metastasis, stage, status of whether have or not have inclusion bodies or psammoma bodies and mitotic level.Correlations were observed between acinar and papillary adenocarcinoma subtypes and EGFR mutations according to the new classification.EGFR mutation was rare in the subtype of solid adenocarcinoma with mucin production and almost never observed in special subtypes (mainly mucinous and colloid adenocarcinoma).In addition, EGFR mutation was associated with the hobnail cell type.Conclusion Lung adenocarcinomas of predominate acinar and papillary histological subtypes with hobnail cell morphology are good predictors for EGFR mutations.

Objective:To understand the willingness to accept peer-referral strategies for promoting HIV testing and related factors in men who have sex with men (MSM) in Shijiazhuang.Methods:A total of 544 MSM were recruited using convenient sampling and sharing two-dimensional code of online questionnaire througth MSM social organizations in Shijiazhuang from August to September in 2018. The anonymous online survey were taken by login through the website "jinshuju.com" ( https://im.jinshuju.com/users/sign_in). The information collected included: the demographic and behavioral characteristics, the attitude to HIV testing for partners, and the willingness to accept peer-referral strategies for promoting HIV testing. The socio-demographic characteristics were analyzed by χ 2 test. Univariate and multivariate logistic regression analyses were conducted to identify the related factors associated with willingness. The SAS 9.4 software was used for statistical analysis. Results:A total of 521 MSM completed the survey. Among them 59.50% (310/521) were willing to advise their partners to receive HIV testing, and 90.02% (469/521) were willing to accept the partners' advice of HIV testing. Higher HIV testing frequency for once a year (a OR=2.72,95% CI:1.42-5.20); for once a half year (a OR=5.72, 95% CI:2.97-11.02); for ≥1 time a quarter (a OR=8.76,95% CI:4.56-16.83), enquiring their partners' HIV status (a OR=1.94, 95% CI: 1.15-3.28) and STD history of their partners (a OR=1.83, 95% CI:1.06-3.14) before having sex were the factors positively associated with the willingness to advise partners to receive HIV testing. Discussing HIV testing with partners (a OR=4.43,95% CI:1.87-10.54) was the factor positively associated with the desire to accept the advice of HIV testing from partners, but feeling emotional hurt by the suggestion of HIV testing (a OR=0.35,95% CI:0.15-0.82) was the factor negatively associated with the willingness to accept the advice of HIV testing from partners. Conclusion:To improve the willingess of MSM to advise their partners to receive HIV testing and strengthen self-protection awareness and equal communication skills are essential for the success of peer-referral strategies for promoting HIV testing among MSM.