BACKGROUND:With the deepen understanding on the biological function of Rho/ROCK pathway, new ROCK inhibitors continue to be discovered, and ROCK inhibitors show good promoting effects on the survival, proliferation and migration of keratocytes. Research on ROCK inhibitors wil provide more donor materials or seed cels for regenerative medicine and clinical cel transplantation. OBJECTIVE:To summarize and explore the progress in the treatment and application of corneal disease using the ROCK inhibitors Y-27632 and Y-39983. METHODS:The PubMed database and CNKI database were retrieved by computer to search the relevant literature published between 2008 to 2015 using the key words of “corneal endothelial cel, corneal epithelial cel, ROCK inhibitor, Y-39983, Y-27632” in English and Chinese, respectively. Relevant articles in line with the theme were screened and analyzed. RESULTS AND CONCLUSION:Totaly 264 papers were initialy searched. At last, 45 papers were selected. Currently there are two main ROCK inhibitors: Y-27632 and Y-39983, but both of which are stil in basic research stage and clinical testing stage. Y-27632 promotes the proliferation and activity of corneal epithelial stem cel after resuscitation; Y-39983 as a novel ROCK inhibitor can be better to inhibit Rho kinases activity than Y-27632, thereby more effectively promoting the healing of the corneal endothelium. There are many studies on the application of ROCK inhibitors in corneal treatment, but not a stable method established to obtain seed cels. Each method has its own advantages and disadvantages, and how to overcome these disadvantages and to find fast and stable access to seed cels is the future direction of development.

Objective To construct a recombinant adeno-associated virus(rAAV)vector containing a human anti-epidermal growth factor receptor(anti-EGFR)single-chain variable fragment antibody gene,and observe its inhibitory effects on pancreatic cancer cell lines.Methods Human anti-EGFR single-chain variable fragment antibody gene was inserted into the Kpn I and Bgl Ⅱ sites to construct a rAAV-anti EGFR vector,and then rAAV1-EGFP group and rAAV1-anti EGFR group were established.The expression of anti-EGFR antibody was observed.Antibody expression was detected by Western blot,and the inhibition and apoptosis rates of human pancreatic cancer cell lines(PCT-3,SW1990,Capan-1,ASPC-1,MiaPaCa-2 and PANC-1 cells)were detected by CCK-8 assay and flow cytometry,respectively.All data were analyzed using the t test.Results The results of Western blot assay demonstrated that anti-EGFR antibody was expressed in 6 pancreatic cancer cell lines.The inhibition rates of rAAV1-EGFP and rAAVl-anti EGFR on pancreatic ASPC-1 cells were 1.1%± 2.4% and 15.1%±3.5%,respectively,with a significant difference between the 2 groups(t =6.598,P ＜0.05).The apoptosis rates of PANC-1 cells were 7.0% ± 3.0% in the rAAV1-EGFP group and 1 1.4% ± 2.5% in the rAAV1-anti EGFR group,with no significant difference between the 2 grouvs(t = 1.952,P ＞0.05).The apoptosis rates of SW1990,ASPC-1,Capan-1,PCT-3,MiaPaCa-2 cells were 1.1% ± 0.8%,1.5% ± 0.7%,1.7% ± 1.2%,1.1%±0.7% and 2.2% ± 1.1% in the rAAV1-EGFP group,and 17.6% ± 2.2%,46.9% ± 3.9%,20.0% ±2.8%,12.1% ± 1.6% and 31.1% ±2.5% in the rAAV1-anti EGFR group,respectively,with significant differences between the 2 groups(t = 12.208,19.846,10.405,10.909,18.327,P ＜0.05).Conclusions A rAAV-anti EGFR vector with human anti-EGFR single-chain variable fragment antibody gene was constructed.Anti-EGFR antibody has obvious inhibition effects on pancreatic cancer cell lines.

Objective EGFR targeted therapy mediated by adeno-associated virus is a promising way to treat pancreatic cancer.This study aimed to assess the feasibility and activity of combining rAAV-anti EGFR,gemcitabine,and radiation in pancreatic cancer cells.Methods Aspc-1 human pancreatic carcinoma cells were divided into several groups,in vitro and in vivo,which were respectively exposed to gemcitabine alone,radiation alone,rAAV-anti EGFR alone,the combination of rAAV-anti EGFR with gemcitabine,the combination of rAAV-anti EGFR with radiation,and the combination of all three agents.The pancreatic cancer tumor growth and apoptotic rate were measured.Results The apoptotic rate was higher in cells treated with a single or combination of agents compared to the negative control (P＜0.05).The combination of rAAV-EGFR,gemcitabine,and radiation produced the highest induction of apoptosis compared to a single agent alone (P ＜ 0.05).Treatment with rAAV-anti EGFR greatly inhibited growth in the tumor xenografts (P＜0.05),and a synergistic effect of rAAV-anti EGFR,gemcitabine,and radiation was found.The number of tissue cancer cells that expressed cleaved caspase-3 after treatment with rAAV EGFR was more than that of the control group (P＜0.05).The combined treatment of rAAV-anti EGFR,gemcitabine,and radiation induced the highest numbers of cells expressing cleaved caspase-3 compared to that with a single agent alone (P＜0.05).Conclusions The rAAV-anti EGFR therapy in combination with chemotherapy and radiation therapy demonstrated a greater efficacy over therapy with a single agent alone.rAAV-anti EGFR increased the efficacy of gemcitabine and radiation in the treatment of pancreatic cancer cells.

Objective:To investigate the immunogeneicity of a subunit vaccine of capsid protein precursor(P1) of swine vesicular diseas(SVD).Methods:In this study,the guinea pigs were immunized with the home-made antigen,T-lymphocyte proliferation response,blocking ELISA and micro-neutralization assay were used to detect the effect of the immunized responses in guinea pigs.Results:The results indicated that a retroviral-based vaccine carrying the capsid protein precursor(P1) of SVD was able to elicit strong SVDV-specific humoral immune responses in guinea pigs.Conclusion:It encourages further work towards the development of a vaccine against SVDV infection.

OBJECTIVE To investigate the effect of clopidogrel(Clog),a platelet aggregation inhibitor,on the development of colitis-associated colon cancer(CAC)and its possible mechanism. METHODS To establish a CAC model,male BALB/c mice were treated with single azoxymethane(AOM) 10 mg · kg-1 by ip. One week later,the mice drank 2.5% dextran sulfate sodium(DSS)for one week and water for two weeks,which lasted three cycles. From the first day mice received 2.5%DSS water, Clog 12.5,25.0 and 50.0 mg · kg-1 was ig administered once a day. Body mass,clinical symptoms,the number of colon tumor and tumor size in colon tissue were recorded. Hyperplasia of tumors was analyzed by HE staining. In the early inflammatory phase of the CAC model,the length of colons was measured, histological structure and epithelium cell proliferation of colon tissues were evaluated by HE staining and Ki67 staining,respectively. In the tumorigenesis and progression phase of the CAC model,epithe?lium cell proliferation of colon tissues was evaluated by Ki67 staining. The mRNA expression of tumor necrosis factor-α(TNF-α)was detected by real-time quantitative PCR. The expression of chemokine(C-X-C motif)ligand 2(CXCL2)and its receptor 2(CXCR2)in colon tissues was detected by PCR and immu?nohistochemistry. RESULTS Compared with model group,clinical symptoms of mice in Clog 12.5 mg · kg-1 group were alleviated,the size of colon tumors was decreased(P<0.05),and hyperplasia of tumors was reduced(P<0.05). During the inflammatory phase,the clinical symptoms of mice in Clog 12.5 mg·kg-1 group were significantly alleviated(P<0.05),the decrease of body mass was reduced(P<0.01),the colon shrinkage was ameliorated(P<0.01),the inflammatory injury and epithelium cell proliferation in colon tissues were reduced(P<0.05). During the tumorigenesis and progression phase,epithelium cell prolif?eration in colon tissues in Clog 12.5 mg·kg-1 group was reduced(P<0.01),and the mRNA and protein expression of TNF-α,CXCL2 and CXCR2 of colon tissues was decreased(P<0.05). CONCLUSION Clog can alleviate inflammation during the CAC early inflammatory phase and inhibit the formation of CAC. The antitumor effect of Clog may be related to the decrease in expression of CXCL2 and CXCR2.

Purpose: This study was to investigate the work experience of newly recruited male nurses during the COVID-19 pandemic. Methods: With a phenomenological approach, this qualitative study was adopted semistructured interviews by phone or video calls. A total of 9 male nurses newly recruited for the COVID-19 wards in Chinese hospitals were interviewed for this study. And Colaizzi's method was applied for evaluation in the data analysis. Results: Based on our findings, three themes were extracted. First, the newly recruited male nurses showed negative emotions at the beginning of COVID-19 epidemic, which was caused by changes in working conditions and content, but also prompted the nurses to change the way of coping with the crisis. Second, they gradually mastered the working skills and psychological training to cope with COVID-19 and developed a positive attitude toward life and a high sense of professional responsibility. Finally, we learned about their needs to respond to public health emergencies such as the COVID-19 pandemic. Conclusion COVID-19 is a disaster for all of humanity. The newly recruited male nurses are an important force in emergency rescue. Although they suffered from short-term negative emotions, they quickly adapted to the crisis. In order to better prepare for future emergencies, the disaster response capacity of newly recruited male nurses needs to be further improved. In addition, newly recruited male nurses have a strong demand for timely and personalized career development guidance.

There are two major problems in the clinical application of Traditional Chinese Medicine injection: inadequate instruction for clinical medication and unreasonable clinical application. The key of the problem is the lack of precise and evidence-based research. In view of the current problems, we should adopt the strategy of government drug administration department leading and gradually taking shape that a multi-collaboration platform of medical institutions, researchers and enterprises to jointly carry out precise and evidence-based research. The specific implementation path is the precise clinical positioning, improving monitoring and assessment system of adverse drug reactions. The medical institutions fully put into the evidence. Chinese medicine injection will be clear and specific in clinical location, with clear mechanism. Survival of the fittest enhances the core competitiveness of Chinese medicine injections and makes contributions to the cause of protecting human life and health.

In clinical practice, the application of Traditional Chinese Medicine injections (TCMIs) has several problems, for example, at some time, the components are complicated, the ingredients and pyrogen are uncontrollable, the usage, the dosage, and the drug interactions are unclear and the method of application is also ambiguous and non-standard. The factors have restricted the development of TCMIs. Based on the policies of the TCMI package inserts since 2006, this paper takes consideration of the problems and gives suggestions of making strict, normal and evidence-based TCMI package inserts.

This study introduces 5 methods of causality assessment of adverse drug reactions (ADRs) domestic and overseas. According to the assessment of ministry of public health of China, the key points about the relevance analysis have been illuminated for the post-evaluation of Chinese patent medicine. Under the guidance of the causality assessment, we should describe the ADR reports in detail and further detect the accurate ADR warning signals. In order to supplement the ADR information, more researches concerning the ADR mechanism should be fully explored.

The herb-drug combination is becoming more and more common. The safety evaluation method is a complex evaluation system. This paper analyzed Pubmed database from 2008 to 2018, and summarized the characteristics of safety evaluation method, and the characteristics are listed as below: (1) Exposure safety issues in real world; (2) The combination use is complex and difficult to evaluate. There are the existing safety assessment methods including literature induction, prospective test, questionnaire survey, etc. Although safety evaluation has been carried out from different angles, there is still a lack of comprehensive and reasonable safety evaluation method. The team tried to combine clinical phenotype and mechanism characteristics to provide more reasonable methodological support for clinical safety evaluation.