1.Analysis of T7 RNA Polymerase: From Structure-function Relationship to dsRNA Challenge and Biotechnological Applications
Wei-Chen NING ; Yu HUA ; Hui-Ling YOU ; Qiu-Shi LI ; Yao WU ; Yun-Long LIU ; Zhen-Xin HU
Progress in Biochemistry and Biophysics 2025;52(9):2280-2294
T7 RNA polymerase (T7 RNAP) is one of the simplest known RNA polymerases. Its unique structural features make it a critical model for studying the mechanisms of RNA synthesis. This review systematically examines the static crystal structure of T7 RNAP, beginning with an in-depth examination of its characteristic “thumb”, “palm”, and “finger” domains, which form the classic “right-hand-like” architecture. By detailing these structural elements, this review establishes a foundation for understanding the overall organization of T7 RNAP. This review systematically maps the functional roles of secondary structural elements and their subdomains in transcriptional catalysis, progressively elucidating the fundamental relationships between structure and function. Further, the intrinsic flexibility of T7 RNAP and its applications in research are also discussed. Additionally, the review presents the structural diagrams of the enzyme at different stages of the transcription process, and through these diagrams, it provides a detailed description of the complete transcription process of T7 RNAP. By integrating structural dynamics and kinetics analyses, the review constructs a comprehensive framework that bridges static structure to dynamic processes. Despite its advantages, T7 RNAP has a notable limitation: it generates double-stranded RNA (dsRNA) as a byproduct. The presence of dsRNA not only compromises the purity of mRNA products but also elicits nonspecific immune responses, which pose significant challenges for biotechnological and therapeutic applications. The review provides a detailed exploration of the mechanisms underlying dsRNA formation during T7 RNAP catalysis, reviews current strategies to mitigate this issue, and highlights recent progress in the field. A key focus is the semi-rational design of T7 RNAP mutants engineered to minimize dsRNA generation and enhance catalytic performance. Beyond its role in transcription, T7 RNAP exhibits rapid development and extensive application in fields, including gene editing, biosensing, and mRNA vaccines. This review systematically examines the structure-function relationships of T7 RNAP, elucidates the mechanisms of dsRNA formation, and discusses engineering strategies to optimize its performance. It further explores the engineering optimization and functional expansion of T7 RNAP. Furthermore, this review also addresses the pressing issues that currently need resolution, discusses the major challenges in the practical application of T7 RNAP, and provides an outlook on potential future research directions. In summary, this review provides a comprehensive analysis of T7 RNAP, ranging from its structural architecture to cutting-edge applications. We systematically examine: (1) the characteristic right-hand domains (thumb, palm, fingers) that define its minimalistic structure; (2) the structure-function relationships underlying transcriptional catalysis; and (3) the dynamic transitions during the complete transcription cycle. While highlighting T7 RNAP’s versatility in gene editing, biosensing, and mRNA vaccine production, we critically address its major limitation—dsRNA byproduct formation—and evaluate engineering solutions including semi-rationally designed mutants. By synthesizing current knowledge and identifying key challenges, this work aims to provide novel insights for the development and application of T7 RNAP and to foster further thought and progress in related fields.
2.Clinical remission and transmural healing of ustekinumab in patients with Crohn's disease
Yun WU ; Yalan XU ; Guoyan ZHANG ; Yuanyuan ZHANG ; Junyao WANG ; Peng YOU ; Tao PENG ; Yulan LIU ; Ning CHEN
Journal of Peking University(Health Sciences) 2024;56(2):253-259
Objective:To treat the Crohn's disease(CD)patients with ustekinumab(UST),to eva-luate their clinical and endoscopic remission,and to evaluate their transmural response(TR)and trans-mural healing(TH)condition using intestinal ultrasonography(IUS).Methods:Retrospective analysis was made on patients diagnosed with CD in Peking University People's Hospital from January 2020 to Au-gust 2022,who were treated with UST for remission induction and maintenance therapy.All the patients were evaluated on both week 8 and week 16/20 after treatment,including clinical,biochemical indica-tors,colonoscopy and IUS examination.Results:A total of 13 patients were enrolled in this study,inclu-ding 11 males and 2 females.The minimum age was 23 years,the maximum age was 73 years and the mean age was 36.92 years.All the patients were in the active stage of disease before treatment,and the average Best Crohn's disease activity index(Best CDAI)score was 270.12±105.55.In week 8,the Best CDAI score of the patients decreased from 270.12±105.55 to 133.16±48.66(t=4.977,P<0.001).Eight patients achieved clinical remission while 5 patients remained in the active stage.Nine patients underwent colonoscopy evaluation.The average simple endoscopic score for Crohn's disease(SES-CD)score decreased from 10.71±7.14 before treatment to 6.00±7.81(t=2.483,P=0.048)in week 16/20.Four patients achieved endoscopic remission while 5 patients did not.In week 8,5 pa-tients achieved TR,2 patients achieved TH,the other 6 patients did not get TR or TH.In week 16/20,6 patients achieved TR,3 patients achieved TH while the other 4 patients did not get TR or TH.There was no significant statistical difference in the TR effect of UST between small intestine and colon lesions(Fisher test,P>0.999).The rate of UST transmural response in the patients who had had previous bio-logical agent therapy was lower than those with no previous biological agent therapy,but there was no sig-nificant statistical difference(Fisher test,P=0.491).Conclusion:After treatment of UST,the clinical and endoscopic conditions of the CD patients had been improved,and some patients could achieve clini-cal remission and endoscopic remission.UST had good TR and TH effects on CD.TR might appear in week 8,and the TR effect increased in week 16/20.There was no significant statistical difference in the TR effect between small intestine and colon lesions.TR effect of UST was better in the patients who had no previous biological agent therapy than those who had had other biological agents,but the result had no significant statistical difference.
3.Clinical Study on Oral Use of Jiawei Puji Xiaodu Granules Combined with External Application of Xiaozhong Sanjie Ointment in the Treatment of Acute Tonsillitis in Children
You-Wei SHANG ; Jian-Guo MAO ; Qing CHEN ; Xue-Feng NING ; Chun-Yang MAO
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(3):638-645
Objective To evaluate the clinical efficacy of oral use of Jiawei Puji Xiaodu Granules(mainly composed of Lonicerae Japonicae Flos,Forsythiae Fructus,Taraxaci Herba,Violae Herba,Schizonepetae Herba,Arctii Fructus,Gleditsiae Spina,Paeoniae Radix Rubra,Moutan Cortex,and Phragmitis Rhizoma)combined with external application of Xiaozhong Sanjie Ointment(mainly composed of Scutellariae Radix,Coptidis Rhizoma,Phellodendri Chinensis Cortex,and Gleditsiae Spina,etc.)in the treatment of acute tonsillitis in children,and to observe their effects on the immune function and related inflammatory indexes of the patients.Methods A total of 116 children with acute tonsillitis of heat stagnation in the lung and stomach type were randomly divided into the control group and the observation group,with 58 cases in each group.The control group was treated with Cefixime Dispersible Tablets,while the observation group was treated with Jiawei Puji Xiaodu Granules for oral use and Xiaozhong Sanjie Ointment for external application.Both groups were treated for 14 days and then were followed-up for a period of 6 months.The changes of traditional Chinese medicine(TCM)syndrome scores,white blood cell(WBC)count,T lymphocyte subset CD3+,CD4+,CD8+ and CD4+/CD8+ levels,and serum levels of tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β),interleukin 6(IL-6)and C-reactive protein(CRP)in the two groups were observed before and after the treatment.Moreover,the clinical efficacy and time for the disappearance of clinical symptoms were compared between the two groups,and the occurrence of adverse reactions and the recurrence of tonsillitis in the two groups were monitored at the same time.Results(1)During the trial,there were 8 cases falling off in the control group but none case falling off in the observation group,and eventually 50 cases in the control group and 58 cases in the observation group completed the full course of treatment.(2)After 14 days of treatment,the total effective rate of the observation group was 98.28%(57/58),while that of the control group was 90.00%(45/50).The intergroup(tested by rank sum test)showed that the clinical efficacy of the observation group was significantly superior to that of the control group(P<0.05).(3)After treatment,the time for the disappearance of sore throat,time for the disappearance of purulent spots,time for subsiding fever and time for the tonsils recovering to normal in the observation group were all significantly shorter than those in the control group(P<0.05).(4)After treatment,the scores of primary and secondary symptoms and the overall symptom scores in the two groups were significantly lower than those before treatment(P<0.05),and the reduction of the scores in the observation group was significantly superior to that in the control group(P<0.05).(5)After treatment,the levels of T lymphocyte subset CD3+,CD4+ and CD4+/CD8+ in the two groups were significantly higher(P<0.05)while the level of CD8 + was significantly lower(P<0.05)than those before treatment,and the increase in the levels of CD3+,CD4+ and CD4+/CD8+ and the reduction of the CD8+ level of the observation group were significantly superior to those of the control group(P<0.05).(6)After treatment,the levels of WBC,TNF-α,IL-1β,IL-6 and CRP in the two groups were significantly lower than those before treatment(P<0.05),and the reduction in the observation group was significantly superior to that in the control group(P<0.05).(7)During the treatment period,no skin allergy,nausea,vomiting or other gastrointestinal adverse reactions occurred in the two groups,which showed a high degree of safety.(8)The 6-month follow-up showed that the recurrence rate of tonsillitis in the observation group was 5.17%(3/58),which was significantly lower than that of 24.00%(12/50)in the control group,and the difference was statistically significant(χ2 = 8.330,P<0.05).Conclusion The efficacy of Jiawei Puji Xiaodu Granules combined with Xiaozhong Sanjie Ointment exert notable curative effect for children with acute tonsillitis of heat stagnation in the lung and stomach type.The combined therapy can significantly shorten the duration of the disease,improve the clinical symptoms of the children and effectively reduce the recurrence rate of tonsillitis.The therapeutic mechanism may be related to the enhancement of the immune function and the inhibition of inflammatory response.
4.Study on the Existing State and Optimization Approach for Executing Orthopedic Case Simulation in a Pediatric Specialty Hospital Using CHS-DRG
Xiaoyan GUO ; Guangying GAO ; Ning XU ; Nianying ZHANG ; Na ZHANG ; Chudi YOU ; Jiashuai TIAN
Chinese Health Economics 2024;43(11):21-26
Objective:To guide cost control and the development of orthopedic specialty services in actual payment by thoroughly analyzing the functioning of DRG simulation in the pediatric orthopedic department of the Capital Institute of Pediatrics.Methods:A total of 736 patients from the orthopedic ward of the Capital Institute of Pediatrics were included as study subjects.The four-quadrant grouping method was employed to examine the profitability status of each diagnosis group in detail.A logistic regression model explored the key factors influencing profitability.Results:Congenital illnesses and traumas account for most pediatric orthopedic cases.Benefit groups(2 groups,4.8%),disadvantageous groups(15 groups,35.7%),key groups(8 groups,19.0%),and potential groups(17 groups,40.5%)are found using the Boston Matrix analysis.The profit group depends on the benefit groups(IF19,IH15)for 67.7%of its total revenue.No matter how profitable they are,medical consumables comprise a sizable portion(43%-45%).According to regression analysis,hospital profitability is highly impacted by age,consumable ratio,examination cost ratio,surgery cost ratio,and other cost ratios(P<0.05).The consumable ratio is the most significant negative predictor(β=-2.238).Conclusion:Hospitals need to optimize their cost structure and increase the probability of profitability through refined management of consumables,the four-quadrant analysis method,strengthened cost accounting,and differentiated resource allocation.
5.Research status of non-coding RNA in viral myocarditis
Xiao-Long HE ; Xin-Xin HU ; Fan-Ning WANG ; Wen-Xin WANG ; Guo-Lei ZHOU ; Kang YI ; Tao YOU
The Chinese Journal of Clinical Pharmacology 2024;40(14):2143-2147
Viral myocarditis(VMC)is the leading cause of dilated cardiomyopathy,which can lead to heart failure and sudden cardiac death.With the development of high-throughput sequencing technology,non-coding RNA(ncRNA)plays an important role in the occurrence and development of VMC.ncRNA promotes the occurrence and development of VMC by regulating viral replication,immune cell function,myocardial cell death,myocardial interstitial fibrosis,and other pathological processes.This article reviews the research progress of ncRNA in VMC and provides new ideas for the pathogenesis,diagnosis,and treatment of VMC.
6.Exploring the mechanism of action of acupuncture-targeted therapy for rheumatoid arthritis and identifying candidate traditional Chinese medicines
Mengya LI ; Yue CHEN ; Ning CHENG ; Haixin YOU ; Changsong DING
International Journal of Traditional Chinese Medicine 2024;46(6):764-772
Objective:Exploration on the mechanism of acupuncture-targeted therapy for rheumatoid arthritis and identification of candidate Chinese materia medica.Methods:Targets of acupuncture for RA were collected from related literature, and GSE205962 gene chips were obtained from GEO database. The weighted co-expression network (WGCNA) was constructed by screening differentially expressed genes using R 4.3.1. The collection targets were taken to intersect with the differential genes, and the common genes were obtained for high-throughput GO and KEGG enrichment. The Hub genes were screened by constructing protein interaction networks using the String database and Cytoscape 3.9.0 software. The CIBERSORT algorithm was used to explore the mechanisms of RA immune infiltration, and finally the Coremine Medical database was applied to predict Chinese materia medica.Results:A total of 8 key targets were selected (RHOA, JAK1, FYN, LCK, STAT1, STAT3, EGF, GATA3). The results of immune infiltration analysis showed that various immune cells were closely related to the biological processes of rheumatoid arthritis, including γδ T cells, monocytes, regulatory T cells, CD8 T cells, etc. Saposheikovize Radix, Cinnamomi Ramulus, Anemarrhenae Rhizoma, Dendrobii Caulis, Rehmannine Radix, Lonicerae Japonicae Flos, Pinelliae Rhizoma, Cimicifugae Rhizoma, Poria, Stephaniae Tetrandrae Radix were the 10 kinds of commonly used key Chinese materia medica in the treatment of RA.Conclusions:Acupuncture and moxibustion may treat RA by regulating RHOA, JAK1, FYN, LCK and other core targets. This study predicts that Chinese materia medica can be used as a source of drugs for the treatment of RA, and can provide new ideas for the combination of acupuncture and medicine for the treatment of RA.
7.Study on the Existing State and Optimization Approach for Executing Orthopedic Case Simulation in a Pediatric Specialty Hospital Using CHS-DRG
Xiaoyan GUO ; Guangying GAO ; Ning XU ; Nianying ZHANG ; Na ZHANG ; Chudi YOU ; Jiashuai TIAN
Chinese Health Economics 2024;43(11):21-26
Objective:To guide cost control and the development of orthopedic specialty services in actual payment by thoroughly analyzing the functioning of DRG simulation in the pediatric orthopedic department of the Capital Institute of Pediatrics.Methods:A total of 736 patients from the orthopedic ward of the Capital Institute of Pediatrics were included as study subjects.The four-quadrant grouping method was employed to examine the profitability status of each diagnosis group in detail.A logistic regression model explored the key factors influencing profitability.Results:Congenital illnesses and traumas account for most pediatric orthopedic cases.Benefit groups(2 groups,4.8%),disadvantageous groups(15 groups,35.7%),key groups(8 groups,19.0%),and potential groups(17 groups,40.5%)are found using the Boston Matrix analysis.The profit group depends on the benefit groups(IF19,IH15)for 67.7%of its total revenue.No matter how profitable they are,medical consumables comprise a sizable portion(43%-45%).According to regression analysis,hospital profitability is highly impacted by age,consumable ratio,examination cost ratio,surgery cost ratio,and other cost ratios(P<0.05).The consumable ratio is the most significant negative predictor(β=-2.238).Conclusion:Hospitals need to optimize their cost structure and increase the probability of profitability through refined management of consumables,the four-quadrant analysis method,strengthened cost accounting,and differentiated resource allocation.
8.Study on the Existing State and Optimization Approach for Executing Orthopedic Case Simulation in a Pediatric Specialty Hospital Using CHS-DRG
Xiaoyan GUO ; Guangying GAO ; Ning XU ; Nianying ZHANG ; Na ZHANG ; Chudi YOU ; Jiashuai TIAN
Chinese Health Economics 2024;43(11):21-26
Objective:To guide cost control and the development of orthopedic specialty services in actual payment by thoroughly analyzing the functioning of DRG simulation in the pediatric orthopedic department of the Capital Institute of Pediatrics.Methods:A total of 736 patients from the orthopedic ward of the Capital Institute of Pediatrics were included as study subjects.The four-quadrant grouping method was employed to examine the profitability status of each diagnosis group in detail.A logistic regression model explored the key factors influencing profitability.Results:Congenital illnesses and traumas account for most pediatric orthopedic cases.Benefit groups(2 groups,4.8%),disadvantageous groups(15 groups,35.7%),key groups(8 groups,19.0%),and potential groups(17 groups,40.5%)are found using the Boston Matrix analysis.The profit group depends on the benefit groups(IF19,IH15)for 67.7%of its total revenue.No matter how profitable they are,medical consumables comprise a sizable portion(43%-45%).According to regression analysis,hospital profitability is highly impacted by age,consumable ratio,examination cost ratio,surgery cost ratio,and other cost ratios(P<0.05).The consumable ratio is the most significant negative predictor(β=-2.238).Conclusion:Hospitals need to optimize their cost structure and increase the probability of profitability through refined management of consumables,the four-quadrant analysis method,strengthened cost accounting,and differentiated resource allocation.
9.Study on the Existing State and Optimization Approach for Executing Orthopedic Case Simulation in a Pediatric Specialty Hospital Using CHS-DRG
Xiaoyan GUO ; Guangying GAO ; Ning XU ; Nianying ZHANG ; Na ZHANG ; Chudi YOU ; Jiashuai TIAN
Chinese Health Economics 2024;43(11):21-26
Objective:To guide cost control and the development of orthopedic specialty services in actual payment by thoroughly analyzing the functioning of DRG simulation in the pediatric orthopedic department of the Capital Institute of Pediatrics.Methods:A total of 736 patients from the orthopedic ward of the Capital Institute of Pediatrics were included as study subjects.The four-quadrant grouping method was employed to examine the profitability status of each diagnosis group in detail.A logistic regression model explored the key factors influencing profitability.Results:Congenital illnesses and traumas account for most pediatric orthopedic cases.Benefit groups(2 groups,4.8%),disadvantageous groups(15 groups,35.7%),key groups(8 groups,19.0%),and potential groups(17 groups,40.5%)are found using the Boston Matrix analysis.The profit group depends on the benefit groups(IF19,IH15)for 67.7%of its total revenue.No matter how profitable they are,medical consumables comprise a sizable portion(43%-45%).According to regression analysis,hospital profitability is highly impacted by age,consumable ratio,examination cost ratio,surgery cost ratio,and other cost ratios(P<0.05).The consumable ratio is the most significant negative predictor(β=-2.238).Conclusion:Hospitals need to optimize their cost structure and increase the probability of profitability through refined management of consumables,the four-quadrant analysis method,strengthened cost accounting,and differentiated resource allocation.
10.Study on the Existing State and Optimization Approach for Executing Orthopedic Case Simulation in a Pediatric Specialty Hospital Using CHS-DRG
Xiaoyan GUO ; Guangying GAO ; Ning XU ; Nianying ZHANG ; Na ZHANG ; Chudi YOU ; Jiashuai TIAN
Chinese Health Economics 2024;43(11):21-26
Objective:To guide cost control and the development of orthopedic specialty services in actual payment by thoroughly analyzing the functioning of DRG simulation in the pediatric orthopedic department of the Capital Institute of Pediatrics.Methods:A total of 736 patients from the orthopedic ward of the Capital Institute of Pediatrics were included as study subjects.The four-quadrant grouping method was employed to examine the profitability status of each diagnosis group in detail.A logistic regression model explored the key factors influencing profitability.Results:Congenital illnesses and traumas account for most pediatric orthopedic cases.Benefit groups(2 groups,4.8%),disadvantageous groups(15 groups,35.7%),key groups(8 groups,19.0%),and potential groups(17 groups,40.5%)are found using the Boston Matrix analysis.The profit group depends on the benefit groups(IF19,IH15)for 67.7%of its total revenue.No matter how profitable they are,medical consumables comprise a sizable portion(43%-45%).According to regression analysis,hospital profitability is highly impacted by age,consumable ratio,examination cost ratio,surgery cost ratio,and other cost ratios(P<0.05).The consumable ratio is the most significant negative predictor(β=-2.238).Conclusion:Hospitals need to optimize their cost structure and increase the probability of profitability through refined management of consumables,the four-quadrant analysis method,strengthened cost accounting,and differentiated resource allocation.

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