Objective To construct a recombinant poxvirus vector vaccine, rVTTδTK-RBD, and to evaluate its safety and immunogenicity. Methods The receptor-binding domain (RBD) gene was synthesized with reference to the gene sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and was inserted into the polyclonal site of the self-constructed recombinant plasmid pSTKE, to construct the recombinant poxvirus shuttle vector pSTKE-RBD. This was then transfected into BHK-21 cells pre-infected with the vaccinia virus Tiantan strain (VTT). The recombinant poxvirus rVTTδTK-RBD was successfully obtained after several rounds of fluorescence phage screening. The effect of rVTTδTK-RBD on the body mass of BALB/c mice was detected after immunizing mice by intra-nasal vaccination. The levels of specific and neutralizing antibodies produced by rVTTδTK-RBD on BALB/c mice were analyzed after immunizing mice intramuscularly. The effect of rVTTδTK-RBD on T cell subsets in BALB/c mice was detected by flow cytometry. Results Through homologous recombination, enhanced green fluorescent protein (EGFP) screening marker, and multiple rounds of fluorescent phosphorescence phage screening, a recombinant poxvirus rVTTδTK-RBD, expressing RBD with deletions in the thymidine kinase (TK) gene, was successfully obtained, which was validated by PCR. The in vivo experiments on BALB/c mice showed that rVTTδTK-RBD was highly immunogenic against SARS-CoV-2 and significantly reduced toxicity to the body compared to the parental strain VTT. Conclusion The recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2 is successfully constructed and obtained, with its safety and immunogenicity confirmed through various experiments.
Animals ; Mice ; SARS-CoV-2/genetics* ; COVID-19 ; Vaccines, Synthetic/genetics* ; Genes, Reporter ; Bacteriophages ; Mice, Inbred BALB C

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

AIM:To explore the effect and mechanism of action of the aqueous extract of Fritillaria ussuriensis(FU-AE)against nonalcoholic fatty liver disease(NAFLD).METHODS:The association between Fritillaria ussuriensis Maxir.(FU)and NAFLD was analyzed by network pharmacology.A mouse model of NAFLD was induced in mice by high fat diet(HFD)+10%fructose drinking water,and three doses of Fritillaria ussuriensis aqueous extract were given to the mice for intervention.Colorimetric assay was used for detection of aspartate aminotransferase(AST),alanine aminotrans-ferase(ALT),triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)levels in the serum of experimental mice.Hematoxylin and eosin staining was used to as-sess the pathological and histological changes in the liver of mice and to clarify the anti-NAFLD effect of aqueous extracts of Fritillaria ussuriensis.Liver tissue proteins were extracted,and expression of proteins related to the AMP-activated pro-tein kinase(AMPK)/acetyl-CoA carboxylase(ACC)pathway was detected by Western blot to clarify the mechanism of an-ti-NAFLD action of Fritillaria ussuriensis.The microbial composition of cecum contents was explored using 16S rRNA se-quencing to reveal the modulatory effect of the aqueous extract of Fritillaria ussuriensis on the structure of intestinal flora in mice with nonalcoholic fatty liver disease.RESULTS:Aqueous extract of Fritillaria ussuriensis(high dose)ameliorated exogenous adipocyte infiltration in the liver of mice with NAFLD(P<0.05).AST,ALT,TG,TC and LDL-C levels were significantly decreased(P<0.05)and HDL-C levels were significantly increased(P<0.05)in the high-dose group.Aque-ous extract of Fritillaria ussuriensis(high dose)significantly increased expression of phosphorylated AMPKα,AMPKα,and phosphorylated ACC in the livers of the model mice(P<0.05),significantly reduced expression of ACC(P<0.05),and significantly increased the relative abundance of the potentially beneficial bacteria Faecalibaculum rodentium,Lacto-bacillus johnsonii,Akkermansia muciniphila(P<0.05).CONCLUSION:Aqueous extract of Fritillaria ussuriensis may ameliorate NAFLD in mice by activating the AMPK/ACC pathway and modulating the structure of intestinal flora.

Objective To explore the association between green space and the risk of dyslipidemia. Methods ^“Dyslipidemia^” and ^“ Normalized Difference Vegetation Index (NDVI)^” were used as search terms to search PubMed, Embase, and Web of Science databases for studies up to September 2023. ARHQ statistical assessment and review tool and NOS scale were employed to evaluate the quality of the studies. R 4.3.1 software was used for meta-analysis. Results A total of 11 studies were included, of which 5 cross-sectional studies and 5 cohort studies were rated as ^“high quality^”. The results of meta-analysis showed that an increase in NDVI in some buffer zones was associated with reduced risks of hypercholesterolemia, hypertriglyceridemia, low HDL-C, and high LDL-C, while an increase in NDVI in 100m buffer zone was significantly associated with reduced risks of all these four diseases, with hypercholesterolemia (OR=0.87, P<0.05), hypertriglyceridemia (OR=0.94, P<0.05), low HDL-C (OR=0.95, P<0.05), and high LDL-C (OR=0.87, P<0.05). Sensitivity analysis suggested that the results of most meta-analyses were robust. Conclusion With the increase in green space near residential areas, the risk of dyslipidemia may decrease.

[摘 要] 目的：探讨光甘草定对肺腺癌细胞A549恶性生物学行的影响及其分子机制。方法：常规方法培养A549细胞和人正常肺上皮细胞BEAS-2B，用不同浓度的光甘草定和或顺铂对其进行处理，通过结晶紫染色、CCK-8法检测光甘草定、顺铂对A549、BEAS-2B细胞增殖活力的影响，Transwell小室实验、细胞划痕实验检测光甘草定对A549细胞侵袭、迁移能力的影响，流式细胞术检测光甘草定对A549细胞凋亡的影响，3D超低黏附板培养法培养A549细胞后采用CCK-8法检测光甘草定对A549细胞增殖的影响，WB法检测光甘草定对A549细胞中上皮间质转化（EMT）相关蛋白表达的影响；构建A549细胞移植瘤模型后检测光甘草定、顺铂对移植瘤的生长以及移植瘤组织中EMT相关蛋白表达的影响。结果：光甘草定、顺铂呈剂量依赖性地显著抑制A549细胞的增殖（P<0.05或P<0.01）、细胞迁移（P<0.05或P<0.01）和侵袭能力（P<0.05或P<0.01），光甘草定能诱导A549细胞凋亡（P<0.01），抑制A549细胞中N-cadherin、snail和vimentin蛋白的表达，促进E-cadherin蛋白表达；光甘草定、顺铂均能抑制A549移植瘤的生长，抑制移植瘤组织中Ki67、N-cadherin、snail和vimentin蛋白的表达、促进E-cadherin蛋白的表达。结论：光甘草定可通过抑制A549细胞的增殖、迁移和侵袭，诱导A549细胞凋亡，其机制可能与其抑制细胞的EMT进程而产生抑癌作用有关。

Objective:To investigate the value of radiomics based on three-dimensional high resolution MR vessel wall imaging (3D HRMR-VWI) for identifying culprit plaques in symptomatic patients with middle cerebral atherosclerosis.Methods:The clinical and imaging features of 117 patients (139 middle cerebral artery plaques) with cerebrovascular diseases in Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from October 2018 to October 2020 were respectively reviewed. Stratified random sampling was used to divide 139 plaques into training set (97 plaques) and validation set (42 plaque) at the ratio of 7∶3. The plaques were divided into 69 culprit plaques and 70 non-culprit plaques based on plaque MR features and clinical symptoms. The clinical and imaging characteristics of culprit plaques and non-culprit plaques were compared by independent sample t-test, Mann-Whitney U test and χ 2 test, and factors with significant difference between two groups in univariate analysis were further analyzed by multivariate logistic regression to find out the independent predictors of culprit plaques. Radiomics features were extracted, screened and radiomics model was constructed using pre-and post-contrast 3D HRMR-VWI based on the training set. The combined model was constructed by combining the independent predictors and radiomics model. Receiver operating characteristic curve and area under curve (AUC) were used to evaluate the efficacy of each model, and DeLong test was used to compare the efficacy of different models. Results:Significant difference was found in intraplaque hemorrhage, lumen area of stenosis, stenosis diameter, stenosis rate, plaque burden and enhancement rate between culprit and non-culprit plaques (all P<0.05). Multivariate logistic regression analysis confirmed that only intraplaque hemorrhage was the independent predictor for culprit plaques (OR=7.045,95%CI 1.402-35.397, P=0.018). In the validation set, the AUC of the pre-contrast 3D HRMR-VWI model was lower than that of the post-contrast 3D HRMR-VWI model ( Z=-2.01, P=0.044). The AUC of pre+post-contrast 3D HRMR-VWI model was not significantly different from that of post-contrast 3D HRMR-VWI model ( Z=0.79, P=0.427). The AUC showed no significant difference between combined model and pre+post-contrast 3D HRMR-VWI model ( Z=-0.59, P>0.05). The combined model showed the best performance in predicting culprit plaques of middle cerebral artery (AUC=0.939), with the sensitivity, specificity and accuracy of 95.24%, 76.19% and 85.71%. Conclusion:Radiomics based on 3D HRMR-VWI has potential values in identifying culprit plaques in symptomatic patients with middle cerebral atherosclerosis.

Objective:This study aims to analyze of the quality of diagnosis and treatment data of community medical institutions on the national health information platform in a district of Beijing from the perspective of scientific research informatization, to provide experience and reference for promoting the informatization construction of primary medical units and tapping the scientific research potential of the regional data platform.Methods:Based on the data backup database of the national health information platform in the region, the data quality was analyzed and evaluated mainly in three dimensions: integrity, integration, and consistency.Results:Through the construction of the national health information platform, the district successfully achieved the effective collection of diagnosis and treatment data from community medical institutions, covering the main data such as patients′ basic information, visit information, test information, prescription information, etc. However, the data collected so far were still insufficient in terms of data integration and consistency.Conclusions:A regional medical data center is suggested to construct to break down the barriers between data systems, conduct pre-structuring of diagnosis and treatment data, improve data integration and consistency, and at the same time, carry out effective scientific research prospective design to promote the effective transformation of clinical data to scientific research data.

Objective:To define more information for familial hematuria by genetic screening in a pedigree with familial hematuria.Methods:This was a 4 generation pedigree included 20 family members. The clinical data and laboratory manifestations of the family members were reviewed and collected from medical records. Meanwhile, the peripheral blood samples of 11 family members of the pedigree were collected, and then DNA samples were extracted by salting out method for genetic analysis. For genetic analysis, firstly, three family members including the proband were selected for whole exome sequencing, and the genetic variations were screened according to the sequence variation interpretation guidelines from the American College of Medical Genetics and Genomics (ACMG) for diagnostic sequence interpretation. Then PCR and Sanger sequencing were used to verify the identified pathogenic variants in all family members in the pedigree.Results:In the pedigree, 6 female members had persistent hematuria. Among them, 2 died due to end-stage renal disease, 2 died due to non-renal diseases, and 2 maintained stable renal function. One of the two members with stable renal function was diagnosed as IgA nephropathy by renal biopsy. Moreover, diffuse basement membrane lesions were identified in her renal biopsy sample after the electron microscope examination, which resulted in the suspected diagnosis of Alport syndrome. Genetic testing in this pedigree revealed two novel mutations in COL4A4 gene (NM_000092), c.G446T:p.G149V in exon 7 and c.G1249A:p.G417R in exon 20. Conclusion:Two novel mutations of COL4A4 gene (c.G446T:p.G149V in exon 7 and c.G1249A:p.G417R in exon 20) in a hematuria pedigree are related with phenotype of familial hematuria.

AIM To study the chemical constituents from the rhizomas of Smilax glauco-china Warb.METHODS The n-butanol fraction of ethanol extract of S.glauco-china was isolated and purified by silica,Sephadex LH-20 and semi-preparative column,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Ten compounds were isolated and identified as phenethanol-β-D-gentiobioside (1),2-phenylethyl-O-β-D-xylopyranosyl-(1 →6)-β-D-glucopyranoside (2),phenylethyl D-rutinoside (3),phenylethyl β-D-glucoside (4),hydrangeifolin Ⅰ (5),icariside D1 (6),calophymembranside B (7),2-hydroxyphenol-1-O-β-D-glucopyranosyl-(6 → 1)-α-L-rhamnopyranoside (8),β-sitosterol (9),daucosterol (10).CONCLUSION All the compounds are isolated from this plant for the first time.