To obtain active protein of pIL-18 expression in Lactococcus lactis, and to observe its biological activity, the total RNA was extracted as template from peripheral blood mononuclear cells. Porcine interleukin 18 (pIL-18) was amplified by RT-PCR. The resulting fragment was cloned into pAMJ399 L. lactis vector, and then transformed to L. lactis MG1363 cells by electroporation. Expression of pIL-18 protein was detected by SDS-PAGE and Western-blotting. Bioactivity of the product was tested by pig spleen lymphocyte proliferation test and cytopathogenic effect inhibition assay. The result of Western blotting and bioactivity test shows that the molecular weight of pIL-18 protein was 19 kDa. The react line was observed in both supernatant and precipitated of the recombinant bacteria pAMJ399-pIL18/MG1363. The expressed pIL-18 can promote the proliferation of pig spleen lymphocyte, and significantly inhibit virus multiplication. As conclusion, porcine interleukin-18 was successfully expressed in L. lactis, and the product was biologically active.
Animals ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Electroporation ; Genetic Vectors ; Interleukin-18 ; biosynthesis ; Lactococcus lactis ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Swine

The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.
Animals ; Anti-Bacterial Agents ; biosynthesis ; Lactobacillus ; metabolism ; Lactoferrin ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Swine

BCG Purified Protein Derivative (BCG-PPD)was isolated and purified from BCG Culture filtrate by trichloroacetic acid and ammonium sulfate methods. The purity of BCG-PPD was Similar to PPD-S(international standard) and PPD-C(China), but more than that of PPD-CT68 (Canada)and PPD-RT23(Danish). The Delayed-Type Hyperseusitivity(DTH) to BCG-PPD was more sensitivity than other PPD on BCG vaccinated guinea pigs, but less sensitivity than other PPD on Mycobacterium tuberculosis infected guinea pigs. The conversion rate and induration diameter to BCG-PPD was higher than PPD in 333 of 12 weeks after BCG vaccination newborns, but lower than that of other PPD in 97 tuberculosis patients. It was shown that DTH reaction to PPD was more sensitivity in Mycobacteria homogeneous strain vaccinated individual than Mycobacteria heterogeneous strain vaccinated individual. It was demonstrated that BCG-PPD was better than other PPD on observation conversion rates and induration diameter of BCG vaccinated individual. It maybe help to identification BCG vaccinated or tuberculosis infected with DTH of BCG-PPD and PPD in same individual.

Background and purpose:HPV infection is known as the primary cause of cervical cancer worldwide To investigate high risk type human papillomavirus (HPV) prevalence and incidence rates of cervical intraepithelial neoplasia (CIN) and their screen risk factors in women with different methods of screening.Methods:1137 residents, workers and service women aged 15-59 from Shenzhen city were investigated for cervical cancer in an epidemiology screening study.The high risk types of human papillomavirus of liquid-based cytology samples were tested by hybrid capture 2 (HC-Ⅱ) and liquid-based cytology test (LCT) was also performed at the same time. Women for HPV-positive with LCT ≥ atypical squamous cells of undetermined sign (ASCUS) or HPV-negative with LCT ≥ low grade squamous intraepithelial lesion (LSIL) were biopsied in colposcopy and then were examined by pathology. All data was managed by Foxbase. ?2 test and unconditional Logistic regression model were used for data analysis by SPSS 10.0.Results:1137 women were eligible in our research, the overall rates of HPV infection was 14.0%. HPV detection rates in residents, workers and service women were 14.1%,9.2%,18.9% respectively. HPV detection rates in workers group was significantly lower than that of service women and residents (P

To evaluate the immune responses of recombinant Lactobacillus casei 393 expressing Porcine Epidemic Diarrhea Viral (PEDV) N protein as oral vaccine, n gene of PEDV was subcloned into the expression vector pPG-1, and then transformed into L. casei 393 by electroporation, resulting in recombinant strain pPG-1-n/L, casei 393. The recombinant strains were induced to express interest protein, which was detected by Western blotting, immunofluorescence microscopy and the whole bacteria ELISA. And then BALB/C mice were used as an animal model immunized with recombinant strains by oral administration, and the immune efficacy was analyzed. The recombinant PEDV N protein showed the antigenic specificity, and was located on the bacterial cell walls of pPG-1-n transformed L. casei. The results of ELISA showed that the mice immunized with recombinant strains could produce remarkable special sIgA level in the feces, and high level of anti-PEDV N protein IgG in the serum (P < 0.01), but the induced antibodies in serum did not demonstrated neutralizing effect. Statistical significant difference was observed among the spleen lymphocyte proliferation index (LPI) among the immunization groups of mice and control groups. And there was significant increase. of IFN-gamma and IL-4 contents in the supernatant of spleen cell culture in immunized group. In conclusion, the oral immunizations with recombinant L. casei 393 can induce significant specific mucosal PEDV N-specific IgA response as well as serum IgG responses, and can evoke both mucosal immune and system immune responses.
Administration, Oral ; Animals ; Antibody Formation ; Coronavirus Infections ; prevention & control ; Female ; Immunity, Mucosal ; immunology ; Lactobacillus casei ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; biosynthesis ; genetics ; immunology ; Porcine epidemic diarrhea virus ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Swine ; Viral Vaccines ; administration & dosage ; immunology

Lactoferrin in milk is a multifunctional protein. In addition, lactoferrin has antiviral, antifungal and antiparasitic activity. In this study, the N-terminus from porcine lactoferrin (PLF-N) was designed to express the antimicrobial action of recombinant porcine lactoferrin. We cloned a 1077 bp fragment of the PLF gene from mammary gland tissue of the lactating sow at the third day. Comparing nucleotide sequence with four strains of PLF gene published on GenBank, the homology was more than 99%. With the reference template of the cloned fragment of PLF-N and optimizing codon bias, we synthesized the gene of N-terminus encoding porcine lactoferrin (PLF-NS). The high expression gene of PLF-NS was cloned into the fusion expression vector pET30b and expressed in E. coli BL21 (DE3). After induced with Isopropyl beta-D-1-Thiogalactopyranoside (IPTG), the target fusion protein was successfully expressed and identified in inclusion bodies by SDS-PAGE and Western blotting. The protein had a molecular weight of 42 kDa and accounted for 32% of the total cellular protein. After purification and renaturation, the purity of the expressed protein was 98%. The expressed PLF-NS protein showed obviously antibacterial activity. This method provides an excellent way for high expression of antimicrobial proteins when optimizing codon bias.
Amino Acid Sequence ; Animals ; Anti-Infective Agents ; metabolism ; pharmacology ; Base Sequence ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Lactoferrin ; biosynthesis ; genetics ; pharmacology ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Swine

For ICU patients, a structured dental assessment and recording system can effectively prevent adverse nursing events caused by tooth loss, and eliminate the potential risk of patients' safety. In addition, the structured recording system can effectively speed up nursing document input, facilitate compliance and acceptance of dental assessment improve work efficiency and quality, and eventually improve the informationization management of nursing.

Regular assessment is a key to early detect and treat delirium. However, studies showed poor compliance and accuracy of delirium assessment by nurses. Moreover, delirium was characterized by acute onset and fluctuation of cognition. Therefore, delirium was frequently unrecognized. Postoperative delirium incidence in the intensive care unit was high and associated with poor outcome. Studies demonstrate delirium was preventable. Delirium prediction models were warranted to identify high-risk critically ill patients in order to apply preventive strategies. This article was to provide an overview of postoperative delirium and review postoperative delirium prediction models for surgical patients in the intensive care unit, thereby providing reference for development, validation as well as application of models in China.

0bjective To translate and revise the Assessment of Interprofessional Team Collaboration Scale (AITCS) into Chinese, then to evaluate the reliability and validity of the Chinese version of AITCS. Methods The Chinese version of AITCS was translated from the original one, back-translated and adjusted for cultural adaptation. The reliability and validity were tested among 288 nurses, 81 physicians, 25 respiratory therapists, 10 physical therapists and 3 nutritionists from a tertiary hospital in Hangzhou using convenience sampling. Results The average of scale-level content validity index was 0.98, unanimity of scale-level content validity index was 0.84 and item-level content validity index was 0.89-1.00. Exploratory factor analysis extracted three common factors,which explained 61.427% of the total variance,and each item had high factor loading quality (>0.4). The correlations coefficients between each dimension score and the total score ranged from 0.801 to 0.898 (P<0.05),and the correlations coefficients between each dimension score ranged from 0.607 to 0.698 (P<0.05). The Cronbach α of AITCS was 0.909, split-half reliability was 0.835 and test-retest reliability was 0.763. Conclusion The Chinese version of the AITCS has been proved to be reliable and valid. It is a valuable tool for evaluating interprofessional team collaboration among the health professional providers in mainland China.

Objective: To investigate the effect of an information module for delirium assessment in ICU. Methods: The information module for delirium assessment was designed according to the route chart of the Confusion Assessment Method for the Intensive Care Unit(CAM-ICU) information assessment. Totally 70 nurses Sir Run Run Shaw Hospital, Zhejiang University School of Medicine were included in the application of the information module. The compliance and accuracy of the delirium assessment and acceptance of nurses' assessment of information module were compared. Results: The information module for delirium assessment was implanted into the clinical critical information system. The rate of compliance and accuracy of delirium assessed was 55.9%(2 728/4 877), 64.2%(231/360) before the application of information module, and 92.0%(4 360/4 740), 92.5%(333/360) after the application of information module. The difference between two groups were statistically significant (χ²=1 611.531, 85.139, P<0.01). The score of nurses′ acceptance of the input method before and after the application of information module were 69.00±4.00, 100.56±3.50, and the difference were statistically significant (t=-49.70, P<0.01). Conclusions The information module for delirium assessment can simplify the assessment process, improve nurses′ compliance and accuracy to assess delirium, optimize critical care management and worth spreading.