Objective To compare the characteristics of remifentanil and sufentanil for tracheal intubation general anesthesia in gyneco-logical laparoscopic operation. Methods 80 patients received gynecological laparoscopic operation were randomly divided into the remifen-tanil group and the sufentanil group with 40 patients in each group, and they were performed total intravenous anesthesia with remifentanil or sufentanil respectively. Compare the changes of heart rate, mean arterial pressure in the two groups before and after induction, at the time of intubation, pneumoperitoneum, and after the operation. And differences in two groups were recorded in the recovery time, extubation time, postoperative pain in 24 hours, exhaust time and postoperative complications. Results Change trend and magnitude in heart rate, mean ar-terial pressure had no significant difference in the two groups (P>0. 05). The recovery time, extubation time in remifentanil group was shor-ter (P<0. 01), postoperative pain in 24 hours in sufentanil group was more mild (P<0. 01). Exhaust time and postoperative complications of the two groups had no significant difference (P>0. 05). Conclusion Remifentanil and sufentanil used in gynecological laparoscopic op-eration can achieve good effect. and effects of the two drugs on hemodynamics were similar. Remifentanil can recieve better effect in terms of recovery time, while sufentanil has better postoperative analgesia effect.

Objective To investigate the application valve of bFGF to improve the viability of trans-plantation flap in Wistar rat. Methods Forty-eight Wistar rats were randomly divided into two groups based on the age: group A,B(1.5 month old, every group was 12 rats) and group C,D (3 month old, every group was 12 rats). After an ischemic model completed, recombinant bovine basic fibroblast growth factor (rb-bFGF) was given to groups A and C in ischemie zone by vascularization flap injection and the equality of normal saline to groups B and D. 14 days postoperatively, the muscular tissue was sent for histology, the blood vessel density was calculated by image analysis, and positive VEGF was detected by immunohistochem-istry. Results The member of capillaries and positive VEGF were more in group A than that in B, and also were morein group C than D(P < 0.05), but there were not statistic difference between group A and C (P > 0.05). Conclusion Recombinant bovine bFGF can stimulate angiogenesis and improve the ischemie vascu-larization flap of rat, which is not associated with their age.

Phosphoinositide 3-kindase,an important signal transduction molecule in cells,plays a key role in the process of cell survival,proliferation and differentiation.Significant progress has been made in the treatment of cancer with PI3K inhibitors,yet the drug resistance of PI3K inhibitors affects their long-term efficacy in clinical treatment.In order to overcome the drug resistance,a variety of rational combined therapies have been developed.In this paper,the research progress of phosphoinositide-3 kinase inhibitors in combination with other drugs to overcome the drug resistance were reviewed.

Objective To investigate the changes of serum milk fat globule epidermal growth factor 8(MFG-E8) levels in pa-tients with carotid atherosclerosis(CAS) ,and the correlation analysis between MFG-E8 and blood glucose ,blood lipid and inflam-matory factors .Methods From October 2015 to February 2017 ,120 patients with carotid atherosclerosis were selected and divided into control group ,thickening group and plaque group ,40 cases in each group .The levels of blood glucose ,blood lipid ,serum MFG-E8 and other inflammatory factors of the three groups were compared ,and the correlation between serum MFG-E8 and other indica-tors .was analyzed .Results The levels of total cholesterol(TC) ,low-density lipoprotein cholesterol(LDL-C) ,tumor necrosis factor alpha(TNF-α) ,and high sensitive C reactive protein (hs-CRP) in plaque group were significantly higher than those in thickening group and control group ,the difference was statistically significant(P<0 .05);the levels of MFG-E8 and interleukin-10(IL-10) in plaque group were significantly lower than those in thickening group and control group ,the difference was statistically significant (P<0 .05);there was no significant difference in the levels of fasting blood glucose (FPG) ,triglyceride(TG) and high-density lipo-protein cholesterol(HDL-C) between the three groups(P>0 .05) .Serum MFG-E8 was negatively correlated with carotid intima-media thickness ,FPG ,TC ,LDL-C ,TNF-αand hs-CRP(P<0 .05)and positively correlated with TG and IL-10(P<0 .05) ,and there was no correlation between serum MFG-E8 and HDL-C(r=0 .105 ,P=0 .255) .Conclusion Serum MFG-E8 can be used as a new index to assess the severity of CAS and it is worthy of clinical application .

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

After sequencing the Asia 1 foot-and-mouth disease virus (FMDV) (As01 strain), we amplified the two fragments covering the whole genome by overlapping PCR and long PCR. The 5' fragment was 1.8 kb in length including 15Cs, and the 3' fragment was 6.7 kb in length. The two fragments were cloned into the pBluescript SK vector to construct recombinant plasmid pBSAs carrying the full-length cDNA of FMDV As01 strain. The RNA transcript was synthesized in vitro using T7 polymerase and transfected into BHK-21 cells. We observed the typical CPE caused by rescued FMDV. The harvested virus was confirmed to be Asia 1 FMDV by RT-PCR, indirect immunofluorescence assay (IFA) and electron microscope observation. The rescued virus showed a similar pathogenicity in suckling mouse (LD50) compared to its wild-type virus. The infectious cDNA clone of the FMDV As01 strain laid a new ground for further investigation of FMDV virulence determinants and development of novel vaccines against FMD.
3' Untranslated Regions ; 5' Untranslated Regions ; Animals ; Base Sequence ; Cattle ; Cell Line ; Cloning, Molecular ; DNA, Complementary ; genetics ; Foot-and-Mouth Disease ; metabolism ; virology ; Foot-and-Mouth Disease Virus ; classification ; genetics ; growth & development ; Genome, Viral ; Mice ; Molecular Sequence Data ; Plasmids ; RNA, Viral ; genetics ; Transfection ; Virulence ; Virus Replication

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

Objective:To explore the distribution features of resident CD8 + T cells infiltration in human esophageal cancer tissues and its clinical significance. Methods:Data from the Cancer Genome Atlas database were retrieved, the correlation between CD103 + CD8 + T cells and infiltration degree of conventional type 1 dendritic cell (cDC1), conventional type 2 dendritic cell (cDC2), type 3 dendritic cell(DC3) was investigated. From January 2006 to December 2008, 78 esophageal cancer tissues and 75 adjacent normal tissues from 78 esophageal cancer patients were collected by Shanghai Outdo Biotechnology Co., Ltd, the clinical data of patients was followed up by telephone until July 2015. The distribution of CD8 + T cells and CD103 + CD8 + T cells in cancer tissues and adjacent normal tissues was detected by multi-color labeling techniques and multispectral tissue imaging. The differences of the number and the ratio of CD8 + T cells and CD103 + CD8 + T cells in cancer tissues and adjacent normal tissues were compared. The Kaplan-Meier survival curves of patients with tissue infiltration of CD8 + T cells and CD103 + CD8 + T cells at different levels were drawn through the R language " survminer" package, and the best cut-off value was obtained. TNM stage, pathological stage and other clinical parameters of patients with high and low infiltration of CD8 + T cells, CD103 + CD8 + T cells were compared. Wilcoxon rank sum test, chi-square test, log-rank test and Cox proportional risk regression model statistical analysis were used to evaluate the prognostic value of the above indicators. Spearman correlation analysis was used for correlation analysis. Results:In the cancer tissues of patients with esophageal cancer, the infiltration degree of CD103 + CD8 + T cells was positively correlated with the infiltration degree of cDC1 cells, cDC2 cells and DC3 cells ( r=0.67, 0.53 and 0.47, all P<0.001). The percentage of CD8 + T cells in all cells in the whole tissue core of tumor tissues (63.09% (42.14%, 76.21%)) was higher than that of adjacent normal tissues (2.56% (1.68%, 5.38%)), and the difference was statistically significant ( U=41.00, P<0.001). The proportion of CD103 + CD8 + T cells in all cells in the whole tissue core of tumor tissues (7.92% (1.60%, 20.61%)) was higher than that of adjacent normal tissues (0.04% (0.01%, 0.10%)), and the difference was statistically significant ( U=857.50, P<0.001). The percentage of high CD8 + T cells infiltration in esophageal cancer tissues of patients with pathological stage Ⅰ+ Ⅱ was lower than that of patients with stage Ⅲ+ Ⅳ (57.9%, 33/57 vs. 85.7%, 18/21); the percentage of high CD103 + CD8 + T cells in CD8 + T cells in esophageal cancer tissues of patients with TNM stage Ⅰ+ Ⅱ was lower than that of patients with stage Ⅲ+ Ⅳ (21.6%, 8/37 vs. 48.8%, 20/41), and the differences were both statistically significant ( χ2=5.25 and 6.23, P=0.022 and 0.013). The results of Kaplan-Meier survival analysis and univariate Cox proportional risk regression model showed that the overall survival (OS) of patients with high CD8 + T cell infiltration was longer than that of patients with low CD8 + T cell infiltration ( HR=0.57, 95% confidence interval (95% CI) 0.34 to 0.96, P=0.034). There was no significant difference in OS between patients with high CD103 + CD8 + T cell infiltration and patients with low CD103 + CD8 + T cell infiltration ( HR=0.66, 95% CI 0.40 to 1.08, P>0.05). Conclusion:The high infiltration of CD103 + CD8 + T cells in esophageal cancer tissues are expected to be used as a prognostic predictor for patients with esophageal cancer, which is an important component of anti-tumor immune response in tumor microenvironment of esophageal cancer.

In order to understand the current development of cell transformation assay (CTA) and its application in the evaluation of cigarette smoke carcinogenesis, the relevant literatures were analyzed and combed from these two aspects. CTA can evaluate the carcinogenicity of various genotoxic and non-genotoxic carcinogens in a short period of time, and has a strong consistency with the results of animal carcinogenic test. After malignant transformation, the cells show changes in cell morphology, immortalization of cells, disappearance of cell-cell contact inhibition, and the ability to form tumors when injected into animals. The identification methods of transformed cells include transformed cell focus count, agglutination test, soft agar culture and inoculation of nude mice, etc. At present, BALB/c 3T3 cells, Bhas 42 cells and SHE cells are the most widely used cells for CTA. Cigarette smoke is a complex aerosol containing a variety of non-genetic carcinogenic chemicals. Cell transformation tests are often used as an in vitro alternative method to evaluate the carcinogenic effects of cigarette smoke, which is different from the short-term genetic toxicity test. It simulates the long-term state of human smoking induced malignant transformation of cells, through the long-term exposure of cells for several decades, which is closer to the occurrence of cancer caused by human smoking. Therefore, CTA can evaluate the carcinogenicity of cigarette smoke and other tobacco products.

Objective: To evaluate the caries status of a cohort of 3-year-old caries-free children from 2 kindergartens in Beijing in a period of 2 years by using Cariostat caries activity test and to assess the sensitivity and specificity of Cariostat score as a caries risk indicator for caries-free children. Methods: Totally 426 3-year-old caries-free children from 2 kindergartens in Beijing were recruited in the present study. Informed consents were obtained from the children's parents. Dental plaque samples of the children were collected and the Cariostat caries activity tests were conducted at baseline and once a year for 2 years. After two years, the caries status of the cohort children were re-evaluated and the caries incidences amongst children with high (2.0, 2.5, 3.0), medium (1.5) and low (1.0, 0.5, 0.0) levels of Cariostat scores were compared and analyzed. Results: Totally 864 3-year-old children from 2 kindergartens were screened before the study startedand 426 (49.3%) children were caries free. After 2-year follow-up, 312 out of 426 (73.2%) remained in the study. The overall caries incident rate was 46.5% (145/312). The caries incident rate of children with high level of Cariostat scores was 88.9% (88/99), while the caries incident rates of children with medium and low levels of Cariostat scores was 38.7% (36/93) and 17.5% (21/120), respectively. The sensitivity and specificity of the Cariostat test in assessing the caries risk of 3-year-old caries-free children in a period of 2 years were 60.7% and 93.4%, respectively. Conclusions Cariostat caries activity test can be used as an indicator to predict the caries risk of 3-year-old caries-free children. Comprehensive caries management could be conducted for children in kindergartens based on the caries risk assessment results of caries experience and the Cariostat score.