Objective:To investigate the prognostic factors of elderly patients with epithelial ovarian cancer(EOC),construct and validate a nomogram prediction model,and provide a basis for clinical diagnosis and treat-ment.Methods:A total of 13128 elderly patients pathologically diagnosed with EOC from 2010 to 2019 in the U.S.SEER database(version 8.4.1)were selected as internal validation data.They were randomly divided in a 7∶3 ratio,with 9138 cases in the training set and 3990 cases in the validation set.All factors were subjected to univari-ate Cox regression analysis;multivariate Cox regression analysis was performed for factors with P＜0.05 to ob-tain their independent prognostic risk factors,and a nomogram model for evaluating 1-,3-,and 5-year overall sur-vival rates was constructed,followed by internal validation.At the same time,the clinical data of 73 elderly EOC patients treated in the First Department of Gynecology at the Affiliated Hospital of Chengde Medical University from January 1,2016 to December 31,2022 were selected for external validation of the nomogram.After construc-ting the nomogram model,by obtaining the risk scores of each independent prognostic factor,the training set,vali-dation set,and external validation set were divided into low-and high-risk groups according to comparison of indi-vidual risk scores and the overall median,the K-M curves for different risk groups were plotted based on the medi-an survival time.Results:①Univariate and multivariate Cox regression analyses showed that,age,marital status,histological type,International Federation of Gynecology and Obstetrics(FIGO)stage,differentiation degree,ser-um tumor carbohydrate antigen 125(CA125)level,unilateral of bilateral tumor,tumor size,positive lymph nodes,residual disease size after cytoreductive surgery,postoperative chemotherapy and surgical treatment were inde-pendent influencing factors for elderly women with EOC(P＜0.05).②According to the drawn column line dia-gram model,the C index values of the training set,the verification set and the external verification set were:0.750,0.732 and 0.798 respectively;the 5-year Area under the curve(AUC)in the Receiver Operating Characteristic(ROC)curve of the external verification set in the three groups was 0.669,while the AUC values for 1-,3-,and 5-year(except the 5-year of external validation)were all greater than 0.700.③Based on the plotted K-M curves,the median survival time of the high-risk groups in the training set,validation set,and external validation set was 31,32 and 39 months respectively,and more than half of the patients in the low-risk group were still alive.The a-bove results all suggested that the column chart model had a high clinical application value for the prediction the 1-,3-,and 5-year overall survival rates of elderly patients with EOC.Conclusions:The nomogram model in this study can accurately evaluate the overall survival rate of elderly with EOC,and provide a basis for individualized treatment.

Objective:To investigate the prognostic factors of elderly patients with epithelial ovarian cancer(EOC),construct and validate a nomogram prediction model,and provide a basis for clinical diagnosis and treat-ment.Methods:A total of 13128 elderly patients pathologically diagnosed with EOC from 2010 to 2019 in the U.S.SEER database(version 8.4.1)were selected as internal validation data.They were randomly divided in a 7∶3 ratio,with 9138 cases in the training set and 3990 cases in the validation set.All factors were subjected to univari-ate Cox regression analysis;multivariate Cox regression analysis was performed for factors with P＜0.05 to ob-tain their independent prognostic risk factors,and a nomogram model for evaluating 1-,3-,and 5-year overall sur-vival rates was constructed,followed by internal validation.At the same time,the clinical data of 73 elderly EOC patients treated in the First Department of Gynecology at the Affiliated Hospital of Chengde Medical University from January 1,2016 to December 31,2022 were selected for external validation of the nomogram.After construc-ting the nomogram model,by obtaining the risk scores of each independent prognostic factor,the training set,vali-dation set,and external validation set were divided into low-and high-risk groups according to comparison of indi-vidual risk scores and the overall median,the K-M curves for different risk groups were plotted based on the medi-an survival time.Results:①Univariate and multivariate Cox regression analyses showed that,age,marital status,histological type,International Federation of Gynecology and Obstetrics(FIGO)stage,differentiation degree,ser-um tumor carbohydrate antigen 125(CA125)level,unilateral of bilateral tumor,tumor size,positive lymph nodes,residual disease size after cytoreductive surgery,postoperative chemotherapy and surgical treatment were inde-pendent influencing factors for elderly women with EOC(P＜0.05).②According to the drawn column line dia-gram model,the C index values of the training set,the verification set and the external verification set were:0.750,0.732 and 0.798 respectively;the 5-year Area under the curve(AUC)in the Receiver Operating Characteristic(ROC)curve of the external verification set in the three groups was 0.669,while the AUC values for 1-,3-,and 5-year(except the 5-year of external validation)were all greater than 0.700.③Based on the plotted K-M curves,the median survival time of the high-risk groups in the training set,validation set,and external validation set was 31,32 and 39 months respectively,and more than half of the patients in the low-risk group were still alive.The a-bove results all suggested that the column chart model had a high clinical application value for the prediction the 1-,3-,and 5-year overall survival rates of elderly patients with EOC.Conclusions:The nomogram model in this study can accurately evaluate the overall survival rate of elderly with EOC,and provide a basis for individualized treatment.

Objective:To express and block the VP7 protein of human group H rotavirus (group H rotavirus, RVH) J19, and to prepare the polyclonal adsorbent polyantibody function of VP7 for identification.Methods:The VP7 protein of human H rotavirus J19 strain was expressed by baculovirus system, and the protein was blocked by affinity chromatography and hydrophobic chromatography. The VP7 polyantiserum was prepared by immunizing the VP7 protein of human H group H rotavirus J19, and the VP7 polyantiserum was identified by ELISA, Western blot (WB) and immunofluorescence.Results:Soluble J19 VP7 protein was obtained. J19 VP7 rabbit polyclonal antibody was prepared. ELISA showed that VP7 polyclonal antibody had a good binding effect, binding to human RVH VP7 protein at 2 048 000-fold. WB showed that the polyclonal antibody could bind to J19 VP7 protein; Immunofluorescence assay showed that VP7 polyclonal antibody could specifically recognize human group H rotavirus J19.Conclusions:The J19 VP7 protein of human H rotavirus was successfully obtained, and the prepared VP7 polyclonal antibody could recognize human H rotavirus, which provided a basis for the detection of human H rotavirus and the study of the structure and function of the viral capsid.

Objective:To express and block the VP7 protein of human group H rotavirus (group H rotavirus, RVH) J19, and to prepare the polyclonal adsorbent polyantibody function of VP7 for identification.Methods:The VP7 protein of human H rotavirus J19 strain was expressed by baculovirus system, and the protein was blocked by affinity chromatography and hydrophobic chromatography. The VP7 polyantiserum was prepared by immunizing the VP7 protein of human H group H rotavirus J19, and the VP7 polyantiserum was identified by ELISA, Western blot (WB) and immunofluorescence.Results:Soluble J19 VP7 protein was obtained. J19 VP7 rabbit polyclonal antibody was prepared. ELISA showed that VP7 polyclonal antibody had a good binding effect, binding to human RVH VP7 protein at 2 048 000-fold. WB showed that the polyclonal antibody could bind to J19 VP7 protein; Immunofluorescence assay showed that VP7 polyclonal antibody could specifically recognize human group H rotavirus J19.Conclusions:The J19 VP7 protein of human H rotavirus was successfully obtained, and the prepared VP7 polyclonal antibody could recognize human H rotavirus, which provided a basis for the detection of human H rotavirus and the study of the structure and function of the viral capsid.

Natural plant-derived diterpenoids are a class of compounds with diverse structures and functions. These compounds are widely used in pharmaceuticals, cosmetics and food additives industries because of their pharmacological properties such as anticancer, anti-inflammatory and antibacterial activities. In recent years, with the gradual discovery of functional genes in the biosynthetic pathway of plant-derived diterpenoids and the development of synthetic biotechnology, great efforts have been made to construct a variety of diterpenoid microbial cell factories through metabolic engineering and synthetic biology, resulting in gram-level production of many compounds. This article summarizes the construction of plant-derived diterpenoid microbial cell factories through synthetic biotechnology, followed by introducing the metabolic engineering strategies applied to improve plant-derived diterpenoids production, with the aim to provide a reference for the construction of high-yield plant-derived diterpenoid microbial cell factories and the industrial production of diterpenoids.
Diterpenes/metabolism* ; Biotechnology ; Metabolic Engineering ; Biosynthetic Pathways/genetics* ; Plants/genetics* ; Synthetic Biology

Danshen, the dried roots and rhizomes of Salvia miltiorrhiza Bunge (S. miltiorrhiza), is widely used in the treatment of cardiovascular and cerebrovascular diseases. Tanshinones, the bioactive compounds from Danshen, exhibit a wide spectrum of pharmacological properties, suggesting their potential for future therapeutic applications. Tanshinone biosynthesis is a complex process involving at least six P450 enzymes that have been identified and characterized, most of which belong to the CYP76 and CYP71 families. In this study, CYP81C16, a member of the CYP71 clan, was identified in S. miltiorrhiza. An in vitro assay revealed that it could catalyze the hydroxylation of four para-quinone-type tanshinones, namely neocryptotanshinone, deoxyneocryptotanshinone, and danshenxinkuns A and B. SmCYP81C16 emerged as a potential broad-spectrum oxidase targeting the C-18 position of para-quinone-type tanshinones with an impressive relative conversion rate exceeding 90%. Kinetic evaluations andin vivo assays underscored its highest affinity towards neocryptotanshinone among the tested substrates. The overexpression of SmCYP81C16 promoted the accumulation of (iso)tanshinone in hairy root lines. The characterization of SmCYP81C16 in this study accentuates its potential as a pivotal tool in the biotechnological production of tanshinones, either through microbial or plant metabolic engineering.
Humans ; Salvia miltiorrhiza/metabolism* ; Biosynthetic Pathways ; Quinones/metabolism* ; Plant Roots/metabolism* ; Gene Expression Regulation, Plant

OBJECTIVETo study the ethylene responsive element binding protein genes of Salvia miltiorrhiza through bioinformatics and characterization of its tissue expression in regenerated plantlets.

METHODThe ethylene responsive element binding protein genes were obtained by cDNA microarray analyze. BLAST was used for alignment, ORF finder software was used to find open reading frame, Prosite database was used to analyze the protein. Semi-quantitative RT- PCR method was used to detect the gene expression level.

RESULTOne ethylene responsive element binding protein was obtained, named as SmERF. SmERF had an open reading frame of 699 bp with 5'-URT 87 bp and 3'-URT 166 bp. The putative protein SmERF contains a highly conserved ERF/AP2 domain. Semiquantitative RT- PCR illustrated that SmERF was expressed in all tissues such as root, stem and leaf in regenerated shoots, while the expression level was higher in root than in stem and leaf.

CONCLUSIONIt was the first time to obtain ERF gene in S. miltiorrhiza and set a good foundation for its further functional study.

DNA-Binding Proteins ; chemistry ; genetics ; metabolism ; Gene Expression ; Genomics ; Open Reading Frames ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Tertiary ; Salvia ; chemistry ; genetics ; metabolism ; Untranslated Regions