Objective:To compare the efficiency and characteristics of different carrier proteins and signal sequences in delivering antigens into Escherichia coli outer membrane vesicles (OMVs). Methods:The fusion protein F1V, which consisted of the main protective antigen of Yersinia pestis F1 and LcrV, was expressed using the carrier proteins such as cytolysin A (ClyA), outer membrane protein A (OmpA), or β-lactamases (Bla) signal sequence as a carrier protein. The expression, localization, and content of F1V protein in OMVs were compared and analyzed. Results:All three delivery methods successfully incorporated F1V protein into OMVs and localized it on the surface of OMVs. Notably, when OmpA was used as the carrier protein, the F1V fusion protein constituted up to 30% of the total protein in OMVs. The highest yield of OMVs, reaching 4.2 mg/L, was achieved when Bla signal sequence was used as the carrier.Conclusions:There is a significant difference in the efficiency of different carrier proteins in delivering the F1V antigen into OMVs of Escherichia coli. Considering both the yield of OMVs and the proportion of antigen in the total protein of OMVs, the carrier Bla signal sequence demonstrated the highest efficiency in delivering F1V into OMVs, showing a potential for the future development of OMVs-based plague vaccines.

Objective:To develop an air transfer stretcher for severely injured patients,so as to meet the requirements of severely injured patients for rapid transport of aviation medical treatment.Methods:The guide rails of hatch of mainstream civil airliner models were analyzed through investigation.Based on the principles of modularization,integration and intelligence,a highly universal and portable aviation stretcher with the functions of rapid transport and life support was designed and developed.The design of the stretcher was verified through simulation analysis and calculation,as well as tests in laboratory,which should meet the requirements of the standards from China Civil Aviation Regulations(CCAR)-25.Results:The results of simulation analysis and tests of laboratory showed that the aviation transport stretcher,and the strength and stiffness of its structure reached to requirement of CCAR-25 standard,which was suit to the portable aviation evacuation of severely injured patients on civil airliners,and the rapid transfer between air and ground.It has the function of supporting life,and the aviation transport stretcher with high universality and convenience can enhance the safety and rescue capability of aviation evacuation system of medical treatment of aviation.Conclusion:The portable aviation stretcher with high-versatility that was researched and developed by this study can effectively solve the problems of life support,injury monitoring,and emergently rescue and treatment during the transport for severely injured patients,which improve the support ability of air-ground transportation for severely injured patients.

Objective:To compare the efficiency and characteristics of different carrier proteins and signal sequences in delivering antigens into Escherichia coli outer membrane vesicles (OMVs). Methods:The fusion protein F1V, which consisted of the main protective antigen of Yersinia pestis F1 and LcrV, was expressed using the carrier proteins such as cytolysin A (ClyA), outer membrane protein A (OmpA), or β-lactamases (Bla) signal sequence as a carrier protein. The expression, localization, and content of F1V protein in OMVs were compared and analyzed. Results:All three delivery methods successfully incorporated F1V protein into OMVs and localized it on the surface of OMVs. Notably, when OmpA was used as the carrier protein, the F1V fusion protein constituted up to 30% of the total protein in OMVs. The highest yield of OMVs, reaching 4.2 mg/L, was achieved when Bla signal sequence was used as the carrier.Conclusions:There is a significant difference in the efficiency of different carrier proteins in delivering the F1V antigen into OMVs of Escherichia coli. Considering both the yield of OMVs and the proportion of antigen in the total protein of OMVs, the carrier Bla signal sequence demonstrated the highest efficiency in delivering F1V into OMVs, showing a potential for the future development of OMVs-based plague vaccines.

Objective:To develop an air transfer stretcher for severely injured patients,so as to meet the requirements of severely injured patients for rapid transport of aviation medical treatment.Methods:The guide rails of hatch of mainstream civil airliner models were analyzed through investigation.Based on the principles of modularization,integration and intelligence,a highly universal and portable aviation stretcher with the functions of rapid transport and life support was designed and developed.The design of the stretcher was verified through simulation analysis and calculation,as well as tests in laboratory,which should meet the requirements of the standards from China Civil Aviation Regulations(CCAR)-25.Results:The results of simulation analysis and tests of laboratory showed that the aviation transport stretcher,and the strength and stiffness of its structure reached to requirement of CCAR-25 standard,which was suit to the portable aviation evacuation of severely injured patients on civil airliners,and the rapid transfer between air and ground.It has the function of supporting life,and the aviation transport stretcher with high universality and convenience can enhance the safety and rescue capability of aviation evacuation system of medical treatment of aviation.Conclusion:The portable aviation stretcher with high-versatility that was researched and developed by this study can effectively solve the problems of life support,injury monitoring,and emergently rescue and treatment during the transport for severely injured patients,which improve the support ability of air-ground transportation for severely injured patients.

Objective To This study aims to analyze the airworthiness compliance of transport stretchers for severely injured patients to ensure their safety and effectiveness in aeromedical rescue.Methods Through finite element modeling and analysis,using the analysis/calculation verification method(code MC2)in the Means of Compliance(MC),the structural static strength,stiffness,load and internal force of the transport stretcher were evaluated,and the damage tolerance assessment and vibration analysis of the connecting structure were carried out.Results The analysis results showed that the safety margins of the main structure and various components of the transport stretcher were all greater than zero,meeting the airworthiness design requirements.The damage tolerance analysis of the connecting structure showed that after the stretcher was installed,the aircraft body structure met the design requirements and did not change the inspection threshold value and interval.In addition,the performance analysis results of the transport stretcher in standard and enhanced vibration environments both indicated that the probability of failure was extremely low.Conclusion The transport stretcher meets the DO-160G standard in design and function and meets the requirements of relevant aviation regulations,providing scientific basis and practical guidance for the efficiency and safety of the aeromedical rescue system.

Objective:To elucidate the factors influencing the differences in the survival rates of esophageal squamous cell carcinoma (ESCC) patients between the rural and urban regions in China. Methods:A total of 36,723 ESCC patients derived from the clinical data-bases containing 500,000 esophageal and gastric cardia carcinoma cases (1973-2015) of the Henan Key Laboratory for Esophageal Can-cer Research of the First Affiliated Hospital, Zhengzhou University, were analyzed. Of these patients, 33,625 were from the rural re-gions (91.6%), comprising 20,906 male patients with an average age of 58.98 ± 8.71 years and 12,719 females with an average age of 59.59 ± 8.53 years. The remaining 3,098 were from the urban regions and composed of 2,089 male patients with an average age of 60.84±9.10 years and 1,009 females with an average age of 62.46 ± 9.14 years. All the patients underwent radical esophagectomy, de-tailed histopathological diagnosis, and TNM staging. Chi square test, Kaplan-Meier, Log-rank, and Cox proportional hazards regression model were used to analyze the differences between ESCC patients from rural regions and those from urban regions and among the risk factors in prognosis. Results:Kaplan-Meier and Log-rank analysis results showed that the ESCC patients from the rural regions had significantly higher overall survival than the urban patients (χ2=12.971, P=0.000). Further analysis showed that rural patients≥50 years old and diagnosed with stage IIa and IIb (middle stage) ESCC had higher survival rates than urban patients in males and females (male:χ2=16.188, P<0.001;female:χ2=5.019, P=0.025). However, the survival rates of rural and urban patients with stage 0,Ⅰa,Ⅰb (early stage) and Ⅲa, Ⅲc, and Ⅳ (late stage) were similar (P>0.05). The results of Cox proportional hazards regression model analysis showed that age, gender, and TNM stages were independent risk factors for rural and urban ESCC patients. When the rural and urban ESCC patients were both considered, the Cox proportional hazards regression model analysis results showed that male ESCC patients≥50 years old, urban residence, and TNM stages were independent risk factors. Conclusion:Rural ESCC patients have significantly high-er overall survival than urban patients. Male, age of≥50 years old, urban residence, and TNM stages were independent risk factors for ESCC patient survival.

OBJECTIVE:To optimize extraction technology of saponins from Paris polyphylla. METHODS:Using paris sapo-nin Ⅰ,paris saponin Ⅱand Paridis total saponin as dependent variables,using ethanol volume fraction,extraction time and sol-vent amount as independent variables,through multiple linear regression and binomial fitting,the extraction technology was opti-mized with response surface method and predicted. RESULTS:The optimized extraction technology of saponins from P. polyphylla was as follows as 10-fold of 80% ethanol,2 times reflux extraction,100 min each time. Under the extraction technology,the ex-traction rates of paris saponinⅠwere 85.4%,82.7% and 87.1%;those of paris saponinⅡwere 85.9%,81.3% and 83.6%;and those of Paridis total saponin were 89.5%,92.1% and 90.3%(all RSD<2.0%). Measured value was 0.964 9,predicted value was 0.986 0 and deviation rate was 2.14%. CONCLUSIONS:The central composite design-response surface method is simple and reliable for the optimization of extraction technology of saponins from P. polyphylla.

This study examined the mechanism by which the gastric cancer cells lead to early peritoneal metastasis. HMrSV5 cells, a human peritoneal mesothelial cell line, were co-incubated with the supernatants of gastric cancer cells. Morphological changes of HMrSV5 cells were observed. The cell damage was quantitatively determined by MTT assay. The apoptosis of HMrSV5 cells was observed under transmission electron microscope. Acridine orange/ethidium bromide-stained condensed nuclei was detected by fluorescent microscopy and flow cytometry. The expressions of Bcl-2 and Bax was immunochemically evaluated. The results showed that conspicuous morphological changes of apoptosis were observed in HMrSV5 cells 24 h after treatment with the supernatants of gastric cancer cells. The supernatants could induce apoptosis of HMrSV5 cells in a time-dependent manner. The supernatants could up-regulate the expression of Bax and suppress that of Bcl-2 in HMrSV5 cells. These findings demonstrated that gastric cancer cells can induce the apoptosis of HPMCs through supernatants in the early peritoneal metastasis. The abnormal expressions of Bcl-2 and Bax may contribute to the apoptosis. Anti-apoptosis drugs promise to be adjuvant chemotherapeutic agents in the treatment of peritoneal metastasis of gastric cancer.
Apoptosis ; Cell Line ; Cell Line, Tumor ; Coculture Techniques ; Epithelial Cells/*cytology ; Epithelium ; Peritoneal Neoplasms/pathology ; Peritoneal Neoplasms/*secondary ; Stomach Neoplasms/*pathology

This study examined the mechanism by which the gastric cancer cells lead to early peritoneal metastasis.HMrSV5 cells,a human peritoneal mesothelial cell line,were co-incubated with the supernatants of gastric cancer cells.Morphological changes of HMrSV5 cells were observed.The cell damage was quantitatively determined by MTT assay.The apoptosis of HMrSV5 cells was observed under transmission electron microscope.Acridine orange/ethidium bromidestained condensed nuclei was detected by fluorescent microscopy and flow cytometry.The expressions of Bcl-2 and Bax was immunochemically evaluated.The results showed that conspicuous morphological changes of apoptosis were observed in HMrSV5 cells 24 h after treatment with the supematants of gastric cancer cells.The supernatants could induce apoptosis of HMrSV5 cells in a time-dependent manner.Th esupematants could up-regulate the expression of Bax and suppress that of Bcl-2 in HMrSV5 cells.These findings demonstrated that gastric cancer cells can induce the apoptosis of HPMCs through supernatants in the early peritoneal metastasis.The abnormal expressions of Bcl-2 and Bax may contribute to the apoptosis.Anti-apoptosis drugs promise to be adjuvant chemotherapeutic agents in the treatment of peritoneal metastasis of gastric cancer.

Objective To characterise the alterations of serum autoantibodies for cyclinB1,p62,Koc-IMP1 and survivin in the subjects with esophgeal and gastric cardia carcinoma and precancerous lesion and their expres-sions in the esophageal and gastric cardia cancer tissue.Methods Enzyme-linked immunoassay and tumor-associated antigen mini-array (consisting of five full-length recombinant proteins,including eyefinB1-p62-Koc,IMP1 and Survivin)were applied to determine the serum level of the autoantibodies of these antigens on 376 subjects with e-sephageal and gastric cardia carcinoma and precancerous lesions.At the same time,the expression of these antigens was detected by immunohistochemical method(ABC)on 13 patients with esophageal cancer and 16 with gastric car-dia cancer.Results All of the 5 antigens determined,the linear correlation Was observed for the detection frequency of cyclinB1,IMPI and p62 in esophageal carcinogenesis,and for p62 in gastric cardia multi-stage progression from normal to precancerous and cancerous lesions(P<0.05).The detection rale with single positive antoantibody im-munoreactivity for both esophageal and gastric cardia cancers were low.However.the positive detection mte for both esophageal and gastric cardia cancer increased apparently when the multiple positive markers were combined together for analysis,which increased tO 3～5 and 3～4 folds respectively.Furthermore,the difference in autoantibody immu-noreactive rate was significant with the lesion progressed from mild tO severe precancerous lesions and to cancer both in esophageal and gastric cardia cancers(P<0.05).The positive immunoreactions of the 5 antigens were detected in cancer tissues.The positive immunostaining rates for cyclinB1,Koc,IMP1 and Survivin both in esophageal and gastric cardia cancers were higher compared to their serllin positive rate of autoantibodies I P<0.05).Of the pa-tients with positive immunostaining in the two cancer tissues,the autoantibodies in the serum for the corresponding antigens could be detected in the same patient.Conclusion The production of the tumor-associated autoantibodies is related tO antigens.The screening rate through serum tumor-associated antigen mini-array for the patients with e-sophageal and gastric cardia carcinoma and precancerous lesions has been increased apparently with combined analy-sis of multiple autoantibodies than with single one.