5.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.
6.Application value of hydrogel in image guided radiotherapy for prostate cancer
Xin ZHAO ; Xue-Wu FAN ; Long TIAN ; Yi-Min HU
Medical Journal of Chinese People's Liberation Army 2024;49(1):37-42
Objective To evaluate the application value of hydrogel in image guided radiotherapy(IGRT)for prostate cancer(PCa).Methods Eighty PCa patients in the First Affiliated Hospital of Hebei North University from October 2022 to February 2023 were collected.The patients were divided into experiment group injected with hydrogel(n=33)and control group without hydrogel(n=47)by central random system allocation.The acute and advanced radiation proctitis(RP)incidence rate of the two groups were compared.Patients in experimental group were divided into RP group(n=5)and non-RP group(n=28).The independent risk factors leading to RP were analyzed by using logistic regression for PCa patients.Results In experiment group,12.1%(4/33)of the patients developed acute grade Ⅰ RP,and 3.0%(1/33)developed advanced grade Ⅰ RP.In control group,31.9%(15/47)of the patients developed acute grade Ⅰ RP,and 12.8%(6/47)developed acute grade Ⅱ RP;19.1%(9/47)of patients developed advanced grade Ⅰ RP,4.3%(2/47)developed advanced grade Ⅱ RP,and 2.1%(1/47)developed advanced grade Ⅲ RP.The incidence of acute and advanced RP in experiment group was lower than that in control group(P<0.05).The application of hydrogel effectively reduced rectal toxicity.Age,rectal volume,V70 and V78 were independent risk factors for the incidence of RP in Pca patients(P<0.05).The characteristics of hydrogel injection were not related to the incidence of RP for Pca patients.Conclusions Hydrogel can effectively reduce the rectal toxicity for Pca patients in IGRT and has little impact on the overall treatment.Hydrogel has certain clinical application and promotion value.
7.Efficacy of Wuling capsules combined with low-dose quetiapine in the treatment of oldest-old patients with insomnia accompanied by anxiety and depression
Chuanyun CAI ; Xin HU ; Huajun ZHU ; Wenzhu LIU ; Zixing WU ; Wei JIANG
China Pharmacy 2024;35(12):1517-1521
OBJECTIVE To investigate the efficacy and safety of Wuling capsules combined with low-dose quetiapine in the treatment of oldest-old patients with insomnia accompanied by anxiety and depression. METHODS The clinical data of 96 oldest- old patients (aged≥80 years) with insomnia accompanied by anxiety and depression who were attending outpatient clinics or hospitalized in our hospital from June 2020 to December 2022 were retrospectively analyzed. According to the different drug treatments, the patients were divided into Wuling capsules group (0.99 g, tid, 34 cases), quetiapine group (25-50 mg, qn, 30 cases) and combination group (using Wuling capsules and quetiapine simultaneously, same as the single drug groups, 32 cases). Before and after 8 weeks of treatment, the patients of three groups were compared in terms of photoplethysmography sleep monitoring indexes (total sleep duration, sleep efficiency, sleep latency, and the number of awakening), Pittsburgh sleep quality index (PSQI) score, 14-item Hamilton anxiety scale (HAMA-14) score, 17-item Hamilton depression scale (HAMD-17) score, mini-mental state examination (MMSE) score, and serum levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α); the therapeutic effects and adverse drug reactions of the three groups were observed and compared. RESULTS Compared with before treatment, the total sleep time, sleep efficiency and MMSE scores of the three groups were significantly longer or higher after treatment (P<0.05), while the sleep latency, number of awakening, PSQI score, HAMA-14 score, HAMD-17 score, serum IL-1β and TNF-α levels were significantly shorter or lower after treatment (P<0.05). Sleep latency, number of awakening, HAMA-14 score, HAMD-17 score and serum TNF-α level in quetiapine group were significantly shorter or lower than Wuling capsules group after treatment (P<0.05). Moreover, the above indexes in the combination group were significantly better than quetiapine group and Wuling capsules group (P<0.05). The total effective rate of treatment in the combination 1731786794@qq.com group was significantly higher than the other two groups (P<0.05). There was no significant difference in the incidence of mail:2408797210@qq.com adverse drug reactions among the three groups (P>0.05). CONCLUSIONS The therapeutic effect of Wuling capsules combined with low-dose quetiapine is better than that of individual drugs for oldest-old patients with insomnia accompanied with anxiety and depression, not increasing the risk of adverse reactions.
8.Effect of Intraoperative Multimodal Analgesia on the Early Postoperative Quality of Recovery in End-stage Head and Neck Cancer Patients Undergoing Open Gastrostomy: A Prospective Randomized Controlled Study
Chunhua HU ; Xiaoyan ZHAO ; Lili WU ; Hongya CHEN ; Xin XU ; Guyan WANG
Medical Journal of Peking Union Medical College Hospital 2024;15(2):359-365
9.Mechanism of icariin regulating the NLRP3 inflammasome against cerebral ischemia reperfusion
Qi ZENG ; Yachen WU ; Maohua HU ; Xiaoyun DA ; Yang LIU ; Xin YANG ; Ying DENG ; Ming LIU
Acta Laboratorium Animalis Scientia Sinica 2024;32(1):25-32
Objective To investigate the mechanism of icariin regulating the NLRP3 inflammasome in the treatment of cerebral ischemia-reperfusion injury in rats.Methods A rat model of focal cerebral ischemia-reperfusion was induced using the thread embolism method.At 24 hours post-operation,the rats were randomly allocated into a sham operation group,model group,butylphthalide group(70 mg/kg),ICA-low dose(20 mg/kg),ICA-middle dose(40 mg/kg),and ICA-high dose(80 mg/kg)groups.The corresponding drugs were administered by gavage at 10 mL/kg once a day for 13 consecutive days.One hour after the last administration,neurological function was scored.The cerebral cortex was observed by hematoxylin-eosin(HE)staining.Expression of interleukin(IL)-1β and IL-18 in the cerebral cortex was determined by immunohistochemistry.Expression of NLRP3,ASC,and Caspase-1 in the cerebral cortex was determined by Western Blot.Results In contrast to the sham operation group,there was a notable increase in neural function scores within the model group.The ischemic area around the visible cerebral cortex showed neuron necrosis at various level or glial cell proliferation,and the number of intact neurons was significantly reduced.IL-1β and IL-18 positive cells were significantly increased.Expression of NLRP3,ASC,and Caspase-1 was significantly increased(P<0.01,P<0.05).After treatment with icariin,the neural function score was decreased significantly.The degree of neuronal necrosis in the peri-ischemic area was significantly reduced,and the number of intact neurons was significantly increased.IL-1 β and IL-18-positive cells were decreased significantly.Expressions of NLRP3,ASC,and Caspase-1 were significantly decreased(P<0.01,P<0.05).Conclusions Treatment of cerebral ischemia-reperfusion injury by icariin may be related to regulation of the NLRP3 inflammasome.
10.Evaluation of Hierarchical Diagnosis and Treatment Effect and Influencing Factors of Beijing Medical Alliance
Tong HU ; Caibo WU ; Xin TIAN ; Youqing XIN
Chinese Hospital Management 2024;44(1):47-52
Objective To understand the status quo of hierarchical diagnosis and treatment effect of medical alliance in Beijing and explore the influencing factors.Methods The convenience sampling method was used to select 26 vertical medical alliances,and the weighted TOPSIS method was used in combination with the index system established in the previous study to evaluate the effect of hierarchical diagnosis and treatment.The factors influencing the effect of hierarchical diagnosis and treatment in medical alliances were summarized through interviews with insiders,and the rank sum test was used to explore the factors influencing the effect of hierarchical diagnosis and treatment in medical alliances.Results The medical alliance B,A2 and A3 ranked high,and the implementation effect was relatively good in the four dimensions of"primary care first consultation,dual-way referral,acute and slow treatment,and vertical linkage";The C2,A8 and F2 medical alliances ranked low,and the implementation effect in the dimensions of"dual-way referral","acute and slow treatment"and"vertical linkage"was significantly lower than that of other medical alliances.The analysis results showed that the differences in the support intensity and core hospital level of different medical alliances were statistically significant(P<0.05),which affected the hierarchical diagnosis and treatment effect of medical alliances.Conclusion While strengthening the information construction and improving the initiative of grassroots and the signing rate of family doctors,it is necessary to improve the support of core hospitals to promote the sinking of resources.Core hospitals should optimize resource allocation according to local conditions and promote hierarchical diagnosis and treatment.

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