Objective:To construct a VP8-mRNA vaccine using human rotavirus spike protein VP8 domain as the immunogen and analyze its immunogenicity in mice.Methods:The VP8-mRNA sequence was designed, optimized, and synthesized. The VP8 gene of rotavirus G1P[8] type was used to construct the plasmid pUC57-VP8-Kan-SapⅠ, which was then sequenced. The plasmid confirmed by sequencing was subjected to large-scale amplification and extraction, followed by linearization, in vitro transcription, and capping. The purified capped products were encapsulated with lipid nanoparticles using a microfluidic control apparatus. The encapsulated VP8-mRNA vaccine was administered intramuscularly to mice at 10, 15, and 20 μg. Serum samples were collected for antibody detection by ELISA. Cellular immune responses were detected by flow cytometry and ELISPOT. Statistical analysis was performed using one-way or two-way analysis of variance and Tukey-Kramer test. Results:The encapsulated VP8-mRNA vaccine was rounded and spherical, with a particle size of about 100 nm, a polymer dispersion index of 0.088, and an encapsulation rate of 92.3%. Two doses of VP8-mRNA vaccine immunization could induce a good immune response in mice. The level of IgG antibody induced after immunization in the 15 μg group was comparable to that of the 20 μg group, and there was no statistical difference ( P>0.05), but the antibody levels in the two groups were significantly higher than that in the 10 μg group ( P<0.000 1). VP8-mRNA vaccine could induce neutralizing antibodies against rotavirus G1 and G9 types. The highest level of neutralizing antibodies against rotavirus type G1 was observed in the 15 μg group, which was significantly higher than that in the 10 μg group ( P<0.05). All immunization groups exhibited good neutralizing ability against rotavirus G9 type. The results of ELISPOT showed that lymphocytes from mice in each vaccine group were able to secrete IFN-γ when stimulated with VP8 peptide. Flow cytometry showed that the proportions of CD8 + T cell subsets in the vaccine groups were higher than that in the control group. Conclusion:The VP8-mRNA vaccine has good immunogenicity in mice and can induce good humoral and T-cell immune responses.

Objective:To construct a VP8-mRNA vaccine using human rotavirus spike protein VP8 domain as the immunogen and analyze its immunogenicity in mice.Methods:The VP8-mRNA sequence was designed, optimized, and synthesized. The VP8 gene of rotavirus G1P[8] type was used to construct the plasmid pUC57-VP8-Kan-SapⅠ, which was then sequenced. The plasmid confirmed by sequencing was subjected to large-scale amplification and extraction, followed by linearization, in vitro transcription, and capping. The purified capped products were encapsulated with lipid nanoparticles using a microfluidic control apparatus. The encapsulated VP8-mRNA vaccine was administered intramuscularly to mice at 10, 15, and 20 μg. Serum samples were collected for antibody detection by ELISA. Cellular immune responses were detected by flow cytometry and ELISPOT. Statistical analysis was performed using one-way or two-way analysis of variance and Tukey-Kramer test. Results:The encapsulated VP8-mRNA vaccine was rounded and spherical, with a particle size of about 100 nm, a polymer dispersion index of 0.088, and an encapsulation rate of 92.3%. Two doses of VP8-mRNA vaccine immunization could induce a good immune response in mice. The level of IgG antibody induced after immunization in the 15 μg group was comparable to that of the 20 μg group, and there was no statistical difference ( P>0.05), but the antibody levels in the two groups were significantly higher than that in the 10 μg group ( P<0.000 1). VP8-mRNA vaccine could induce neutralizing antibodies against rotavirus G1 and G9 types. The highest level of neutralizing antibodies against rotavirus type G1 was observed in the 15 μg group, which was significantly higher than that in the 10 μg group ( P<0.05). All immunization groups exhibited good neutralizing ability against rotavirus G9 type. The results of ELISPOT showed that lymphocytes from mice in each vaccine group were able to secrete IFN-γ when stimulated with VP8 peptide. Flow cytometry showed that the proportions of CD8 + T cell subsets in the vaccine groups were higher than that in the control group. Conclusion:The VP8-mRNA vaccine has good immunogenicity in mice and can induce good humoral and T-cell immune responses.

ObjectiveTo analyze the research status and trends in low-level occupational benzene exposure. Methods Articles on low-level occupational benzene exposure from Chinese and English journals from January 1st, 2000, to December 31th, 2022 were retrieved using the Web of Science and the China National Knowledge Infrastructure, and a bibliometric analysis was conducted. Results A total of 327 articles were included in the analysis, comprising 216 English articles and 111 Chinese articles. i) The number of articles published in English fluctuates greatly over the years, without a trend of continuous growth or decline. Authors from 359 research institutions in 45 countries and regions have published relevant English articles in 97 kinds of journals, involving 281 grants from 226 foundations. The top three countries in terms of articles amount were the United States, Italy, and China, with 81, 46, and 43 papers, respectively. The English articles mainly focused on mechanistic research at the genetic level, such as hematotoxicity, oxidative stress, and DNA damage. ii) The number of Chinese articles increased gradually after 2012, with the growth peak in 2017. Authors from 127 research institutions in 26 provinces, autonomous regions, and municipalities published Chinese articles in 51 kinds of journals, involving 154 grants from 78 foundations. Chinese articles tended to focus on benzene-induced hematotoxicity and occupational health damage. Conclusion Most studies on low-level occupational benzene exposure were conducted in China, the United States and Italy, focused on hematotoxicity. Monitoring international research topics and hotspots of the field has certain reference value for related research in China.

Objective:To analyze the influential factors of blurred vision after general anesthesia.Methods:The clinical data of 997 patients who underwent elective general anesthesia at The No. 1 People's Hospital of Pinghu from September 2022 to May 2023 were retrospectively analyzed. The data collected included age, sex, body mass index, American Society of Anesthesiologists classification, history of hypertension, history of diabetes, operation duration (specifically whether it exceeded 3 hours), surgical position (whether the patient was in the supine position), operating room temperature, use of penehyclidine hydrochloride, use of muscle relaxant antagonists, use of atropine, blood pressure (specifically whether it was ≥ 30% of the baseline value), fluid input, blood loss, and use of pneumoperitoneum. Collinearity diagnosis and univariate logistic regression analysis were conducted to select factors with statistical significance. Subsequently, multivariate logistic regression analysis was performed.Results:Univariate and multivariate logistic regression analyses showed that age > 65 years ( OR = 1.47, 95% CI: 1.01-2.15, P = 0.043), surgical position (non-supine position) ( OR = 1.54, 95% CI: 1.06-2.25, P = 0.025), operation time exceeding 3 hours ( OR = 1.76, 95% CI: 1.05-2.94, P = 0.031), and the use of penehyclidine hydrochloride ( OR = 4.91, 95% CI: 3.35-7.21, P < 0.001) were identified as factors contributing to postoperative blurred vision in patients undergoing general anesthesia. Conclusion:Factors contributing to postoperative blurred vision in patients undergoing general anesthesia include age > 65 years, the use of penehyclidine hydrochloride during surgery, operation time exceeding 3 hours, and non-supine surgical position. Clinically, it is essential to implement early and effective preoperative education, enhance intraoperative nursing quality, and optimize preoperative medication for general anesthesia to reduce the incidence of blurred vision after surgery.

Objective To analyze the effects of painting and medication intervention on psycho-logical state,compliance and quality of life in patients with generalized anxiety disorder(GAD).Methods A total of 70 patients with GAD from May 2020 to September 2022 were included as study subjects,among whom 35 patients receiving single drug intervention were included in single group,and 35 patients receiving painting combined drug intervention were included in combined group.The psychological state scores and intervention compliance of the subjects in the two groups were ana-lyzed,the life quality scores and serum factor expression levels of the subjects in the two groups were compared before and after intervention,and the changes in sleep quality of the two groups were ob-served before and after intervention.Results After intervention,the scores of Hamilton Anxiety Rat-ing Scale(HAMA),Hamilton Depression Rating Scale(HAMD)in the two groups were decreased compared with those before intervention,but their scores in the combined group were lower,and the differences were statistically significant(P＜0.05).The total compliance rate of the combined group was higher than that of the single group(P＜0.05).After intervention,the 36-item Shot-form Health Status Survey(SF-36)scores of the two groups were improved compared with that before intervention,and the SF-36 score of the combined group was higher,the difference was statistically significant(P＜0.05).After intervention,the expression levels of brain derived neurotrophic factor(BDNF)and malondialdehyde(MDA)in the combined group were lower than those in the single group,and the expression level of superoxide dismutase(SOD)was higher than those in the single group(P＜0.05).After intervention,the sleep quality of the two groups was improved compared with that be-fore intervention,but the improvement degree of the combined group was higher(P＜0.05).Conclusion Painting combined with drug intervention can effectively improve the adverse psycho-logical state of GAD patients,improve their intervention compliance,and further improve their qual-ity of life and sleep quality.

Objective To analyze the effects of painting and medication intervention on psycho-logical state,compliance and quality of life in patients with generalized anxiety disorder(GAD).Methods A total of 70 patients with GAD from May 2020 to September 2022 were included as study subjects,among whom 35 patients receiving single drug intervention were included in single group,and 35 patients receiving painting combined drug intervention were included in combined group.The psychological state scores and intervention compliance of the subjects in the two groups were ana-lyzed,the life quality scores and serum factor expression levels of the subjects in the two groups were compared before and after intervention,and the changes in sleep quality of the two groups were ob-served before and after intervention.Results After intervention,the scores of Hamilton Anxiety Rat-ing Scale(HAMA),Hamilton Depression Rating Scale(HAMD)in the two groups were decreased compared with those before intervention,but their scores in the combined group were lower,and the differences were statistically significant(P＜0.05).The total compliance rate of the combined group was higher than that of the single group(P＜0.05).After intervention,the 36-item Shot-form Health Status Survey(SF-36)scores of the two groups were improved compared with that before intervention,and the SF-36 score of the combined group was higher,the difference was statistically significant(P＜0.05).After intervention,the expression levels of brain derived neurotrophic factor(BDNF)and malondialdehyde(MDA)in the combined group were lower than those in the single group,and the expression level of superoxide dismutase(SOD)was higher than those in the single group(P＜0.05).After intervention,the sleep quality of the two groups was improved compared with that be-fore intervention,but the improvement degree of the combined group was higher(P＜0.05).Conclusion Painting combined with drug intervention can effectively improve the adverse psycho-logical state of GAD patients,improve their intervention compliance,and further improve their qual-ity of life and sleep quality.

Objective To assess the factors associated with the restoration of spontaneous circulation (ROSC) and 2-year survival prognosis in patients with cardiac arrest (CA) after acute myocardial infarction (AMI),and after ROSC,the effects of various factors on midian survival time and on 2-year survival.Methods In a registry study from January 2005 to January 2015,all consecutive AMI-induced CA patients treated with cardiopulmonary resuscitation (CPR) admitted to our hospital were enrolled.The survivors were followed-up for 2 years.Univariate analysis was applied to evaluate factors associated with rate of ROSC and 2-year survival.Multivariable logistic regression analysis was applied to evaluate statistically significant factors in the univariate analysis.Medians with inter-quartile ranges were used to describe 2-year survival time affected by various factors after ROSC.Kaplan-Meier survival curve analysis was used to evaluate the effect of factors on 2-year survival.Results A total of 254 cases with CA after AMI were enrolled,including 129 cases of ROSC and 71 cases of 2-year survival.Univariate analysis showed age ≥ 70 years,CA occurred during 22:00-8:00,the duration time ofCPR ≥ 15 min and adrenaline dosage ＞ 5 mg were unfavorable predictors of ROSC;while,left ventricular ejection fraction (LVEF) ≥ 40％ before CA,shockable rhythm and percutaneous coronary intervention (PCI) therapy were favorable predictors.Besides,age ≥ 70 years,intubation during CPR,adrenaline dosage ＞ 5 mg and cardiogenic shock were unfavorable predictors of 2-year survival;While,male,normal daily activity before CA and PCI treatment were favorable predictors.Multivariable analysis showed age,the duration of CPR,adrenaline dosage,LVEF before CA,the rhythm during CPR and PCI therapy were independent predictors of ROSC.Age and PCI therapy were independent predictors of 2-year survival.Among patients,the survival time was affected by various factors after ROSC,and the factors with minimum 25％ and small median value were associated with cardiac rupture,cancer,adrenaline dosage ＞ 5 mg and cardiogenic shock.The factor with maximum 25％ value was PCI treatment (216 days).Kaplan-Meier survival analysis suggested that age ≥ 70 years was an unfavorable factor of 2-years survival (Log-rank test,P=0.007);while,PCI treatment was a favorable factor (Log-rank test,P＜0.01).PCI-related prognosis analysis showed that the effectiveness of PCI was related to the timing of PCI,the number of infarctrelated artery and the difference in culprit lesion.Conclusions The age ≥ 70 years was disadvantageous to both ROSC and 2-year survival.PCI treatment was favorable to both ROSC and 2-year survival.

Rhubarb is a traditional Chinese medicines which possess laxative, lipid-lowering, and weight-loss activities, but the active compounds of lipid-lowering and underlying molecular mechanisms are not yet clear. This study aims to explore the effects of chrysophanol on the mRNA expressions of sterol regulatory element-binding proteins (SREBPs) and lipid metabolism in human liver carcinoma Huh-7 cells, which is one of the active compounds obtained from Rhubarb. A reporter gene assay was used to test the transcription of SREBP. The intracellular triglyceride and total cholesterol contents were measured by using commercially available test kits. The SREBPs target genes expressions were measured by Quantitative Real-Time PCR. Cell viability was evaluated by Cell Counting Kit-8. As the results shown, chrysophanol (40 μmol · L(-1), 16 h) could notably inhibited human SRE promoter activity in a dose-dependent manner and decrease intracellular cholesterol and triglyceride levels. Furthermore, the mRNA expressions of SREBPs target genes were significantly downregulated by chrysophanol treatment. However there are no significant differences on cell viability when compared with the control group. These results suggested that chrysophanol might improve lipid metabolism through suppressing the mRNA expressions of SREBPs target genes to attenuate intracellular lipid accumulation.

OBJECTIVETo explore the distribution of the main clinical symptoms and signs of primary biliary cirrhosis (PBC) in a population of individuals with positivity for anti-mitochondrial antibody-M2 (AMA-M2).

METHODSA total of 20 970 persons who participated in routine health examinations at our hospital were tested for presence and level of antinuclear antibodies (ANAs) using an indirect immunofluorescence (IIF) assay and of AMA-M2 by western blotting. The chi-square test was used for statistical analysis.

RESULTSTiters of ANAs more than 1:320 were detected in 1 243 of all the study participants, with 156 of those individuals having detectable AMA-M2.The overall rate ofAMA-M2 positivity was 0.74%, with a significantly higher rate among female subjects (males:0.3% (32/10 550) vs.females:1.2% (124/10 420); x2=55.85, P less than 0.05). Among the AMA-M2-positive population there were 66 cases of abnormal liver function, 58 cases of increased alkaline phosphatase, 72 cases of abnormal findings for routine blood testing, 47 cases of gallbladder disease history, 49 cases of diabetes history, 22 cases of allergy, 75 cases of abdominal discomfort, 38 cases of weakness, 3 cases of jaundice, and 11 cases of pruritus. There were significant differences between the AMA-M2-negative individuals and the AMA-M2-positive individuals.

CONCLUSIONAmong the general population, individuals with substandard states of health, such as those with abnormal findings in routine blood tests and abnormal liver function, should be screened for AMA-M2. This screening will facilitate early diagnosis of PBC and timely initiation of disease management, improving the patient's life quality of life and prolonging their life.

Biomarkers ; Blotting, Western ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; Liver Cirrhosis, Biliary ; metabolism ; Male ; Pruritus ; Quality of Life

Objective To study the distribution of antinuclear antibodies ( ANAs) in a healthy population and the significance of using ANAs screening test in medical examination .Methods The ANAs were measured by indirect immunofluorescence assay ( IIF) .The Western blot assay was used to detect fif-teen specific antibodies against auto-antigens .Results 3519 out of all 25 110 subjects showed ANAs titers>1∶100 , and among them male and female subjects were respectively accounted for 1143 and 2376 .1489 out of all subjects had ANAs titers >1∶320 , and among them male and female subjects were respectively accounted for 406 and 1083 .The positive rates of ANAs at different titers showed significant differences be -tween male and female subjects .Among subjects with ANAs titers >1∶320 , the number of male subjects showed a steady increase with the age , while the percentage of female subjects reached to two peaks during the periods of puberty and menopause .The fifteen specific antibodies were detected in 659 out of 1489 sub-jects with ANAs titers>1∶320 and anti-Ro-52 (14.2%) accounted for the majority , followed by anti-M2 (12.7%) and anti-SSA (9.6%).Conclusion ANAs can be detected among healthy population of all ages, but their distribution varied with gender and age .ANAs screening test is necessary for medical exami-nation of healthy population , especially for female during period of puberty or menopause .The population with positive ANAs should be followed-up closely and educated for the prevention of autoimmune diseases .