Objective To construct a multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector and identify it preliminarily. Methods According to recombinant pcDNA3-p30-ROP2 restriction sites,HBV HBsAg gene sequences of primers were designed and synthesized to amplify target fragment,and then cloned into pcDNA3-HbsAg-p30-ROP2 expression vector. Af-ter sequencing,it was identified finally by restriction enzyme digestion and other molecular biology techniques. Results HBV HBsAg gene segment was amplified by PCR and the multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector was constructed and identified to be correct as theoretical values. The PCR and restriction enzyme digestion results showed that HBsAg and p30-ROP2 gene in recombinant plasmid were confirmed by DNA sequencing. Conclusion The multi-gene recombinant pcD-NA3-HBsAg-p30-ROP2 expression vector is successfully constructed.

Objective To identify the trends of vegetables and fruits consumption among Chinese adults aged 18 to 44 years old from 1991 to 2011.Methods Twenty four hour dietary recall data from China Health and Nutrition Survey (1991,1993,1997,2000,2004,2006,2009 and 2011) were used to identify the trends of vegetables and fruits consumption among Chinese between 18 and 44 years old.Results From 1991 to 2011,the proportion of daily consumption of vegetables rarely varied,while the vegetables intake declined significantly; and the proportion of daily consumption and intake of fruits kept increasing as follows.By 2011,the proportion of daily consumption and intake of vegetables and fruits among the respondents were 99.7％,48.0％,321.6 g/d,and 90.1 g/d respectively.A significant drop was found in the vegetables intake among the respondents against the vegetables intake as recommended by the Chinese dietary guidelines,up to 50.2％ in 2011; a significant rise was found in the fruits intake of the respondents against the fruits intake as recommended by the said guidelines,up to 17.4％ in 2011.Conclusion The average daily intake of vegetables and fruits of young and middle-aged residents (18-44 age group) in nine provinces in China was found lower than that recommended in the Chinese dietary guidelines;in view of the high proportion of people having less vegetables and fruits intake that those recommended by the Chinese dietary guidelines,further measures are expected to encourage their vegetables and fruits intake.

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.

Objective The researchers systematically retrieved,evaluated,and summarized the best evidence for the care of nebulized inhalation in adult patients on mechanical ventilation,to provide a basis for standardizing the care of nebulized inhalation in patients on mechanical ventilation.Methods We systematically searched the domestic and foreign databases to collect the evidence on the literature related to nebulization therapy for mechanical ventilation in adults.The time for the retrieval is from the inception of databases until February 2023.There were 3 researchers who evaluated the quality of the literature,and extracted and summarized the evidence based on this subject.Results A total of 19 articles were obtained in database.42 pieces of evidence were summarized,covering pre-assessment of nebulized inhalation,preparation before nebulized inhalation,medication management,selection and standardized use of nebulization devices,respiratory machine mode and parameter settings,equipment management during nebulized inhalation,evaluation of effect,management of adverse reactions,disposal of materials and environment after nebulized inhalation and management of nebulized inhalation for respiratory infectious diseases.Conclusion This study summarized the best evidence for nebulized inhalation nursing in adult patients with mechanical ventilation,so as to provide a reference of standardized nebulized inhalation therapy for such patients,which is conducive to ensuring the safety of patients.

Objective:To investigate the expression of heterogeneous nuclear ribonucleoprotein A2B1 (HNRNPA2B1) in human esophageal cancer tissues and its clinical significance.Methods:Single-cell data for esophageal cancer were downloaded from the Gene Expression Omnibus (GEO) database (GSE160269 dataset, last updated on November 29, 2020) to analyze the expression of HNRNPA2B1. Transcriptional sequencing data for esophageal cancer from The Cancer Genome Atlas (TCGA) database, including the fragments per kilobase of transcript per million mapped reads (FPKM) quantitative data (173 samples, consisting of 162 esophageal cancer tissues and 11 adjacent normal tissues), and survival data in the phenotype category were downloaded. Analysis of FPKM quantitative data from the TCGA database for esophageal cancer was performed. The top 250 genes most correlated with HNRNPA2B1 were selected and the R4.3.0 clusterProfiler package was used to conduct Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses on the selected gene set. FPKM quantitative data from the TCGA database for esophageal cancer were imported into the CIBERSORTx website to obtain immune cell abundance scores, and the correlation between HNRNPA2B1 and the degree of immune cell infiltration was analyzed. The clinicopathological data of patients from esophageal cancer tissue microarrays including 114 cases of esophageal squamous cell carcinoma tissues and 66 cases of adjacent normal tissues were collected. The patients underwent surgery from January 2006 to December 2008, and the follow-up period extended until July 2015. Cytokeratin (CK) and HNRNPA2B1 expression in esophageal cancer tissue microarrays were detected by using multi-color immunohistochemical (mIHC) staining, and multispectral tissue imaging was conducted. The R4.3.0 survival package and survminer package in TCGA database were used to calculate the optimal cut-off value of HNRNPA2B1 expression and the proportion of CK + HNRNPA2B1 + cells in tissue microarrays was used to calculate the cut-off value of HNRNPA2B1 expression based on which patients were categorized into high and low expression groups. The overall survival (OS) of both groups was compared and the factors influencing OS were analyzed by using the Cox proportional hazards model. Results:In the GSE160269 dataset of single-cell data for esophageal cancer, the expression level of HNRNPA2B1 in tumor epithelial cells was higher than that in normal epithelial cells, and HNRNPA2B1 was highly expressed in various immune cell subtypes. The high expression level of HNRNPA2B1 was positively correlated with regulatory T cells, naive B cells and memory CD4 + T cells. GO enrichment analysis revealed that HNRNPA2B1 was primarily involved in the biological process of nuclear division, cellular components were mainly enriched in chromosomal regions, and molecular functions were mainly enriched in ATP hydrolysis activity. KEGG enrichment analysis indicated that HNRNPA2B1 was primarily involved in biological processes such as the cell cycle, spliceosome, and DNA replication. Results from mIHC and multispectral tissue imaging demonstrated that CK was predominantly expressed in the cell membranes of tumor cells and normal esophageal epithelial cells, while HNRNPA2B1 was primarily expressed in the nuclei of tumor cells and normal esophageal cells. The expression level of HNRNPA2B1 in esophageal cancer tissues was higher than that in the normal paracancerous tissues ( U = 2 984.00, P < 0.05). Results of tissue microarrays and the survival analysis on the data in the TCGA database indicated that esophageal cancer patients with low HNRNPA2B1 expression had a better OS compared to those with high expression (both P < 0.05). Cox multivariate regression analysis revealed that age ( HR = 1.919, 95% CI: 1.158-3.182, P = 0.011), TNM stage ( HR = 2.404, 95% CI: 1.374-4.207, P = 0.002), T stage ( HR = 2.349, 95% CI: 1.150-4.789, P = 0.019), and the expression of HNRNPA2B1 in tumor epithelial cells ( HR = 2.160, 95% CI: 1.280-3.647, P = 0.004) were independent factors influencing OS in esophageal cancer patients. Conclusions:The high expression of HNRNPA2B1 protein in esophageal cancer tissues may play a role in the developement and progression of esophageal cancer, serving as a crucial biological indicator for prognostic assessment of esophageal cancer.

[Abstract] Objective: To investigate the expression of chemokine-like factor-like MARVEL transmembrane domain-containing family member 6 (CMTM6) in breast cancer tissues and its correlation with clinicopathological features and prognosis of patients. Methods:Atotal of 136 breast cancer tissue chips (purchased from Superchip Company), including 42 pairs of matched cancer and paracancerous tissues, were used for this study. The expression level of CMTM6 in cancer and paracancerous tissues was detected by immunohistochemistry. The comparison of CMTM6 expression between breast cancer and paracancerous tissues was conducted by paired χ2 test. The relationship between CMTM6 expression in breast cancer tissues and the clinicopathological characteristics of patients was analyzed by χ2 test. Kaplan-Meier and Log rank test analyses were used to analyze the relationship between CMTM6 expression and the survival of patients, and Cox model was used to evaluate the effect of different indicators on the prognosis of patients. Results: The expression of CMTM6 in breast cancer tissues was significantly higher than that in paracancerous tissues (P<0.01). The expression of CMTM6 was correlated with pathological type of breast cancer and HER2 positivity (P<0.05). The survival time of patients in CMTM6 high expression group was significantly shorter than that of patients in CMTM6 low expression group (P<0.05). Pathological type (HR=10.374, 95%CI: 3.529-30.497, P<0.01), TNM stage (HR=4.599, 95%CI: 1.784-11.856, P<0.01), triple-negative breast cancer (HR=3.370, 95%CI: 1.055-10.761, P<0.05) and high expression of CMTM6 (HR=0.195, 95%CI: 0.073-0.518, P<0.01) were independent risk factors for prognosis of breast cancer patients. Conclusion: CMTM6 is highly expressed in breast cancer tissues, which can be used as a risk factor for prognosis evaluation of breast cancer patients.