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.Palatability evaluation of oral iron for children based on in vivo and in vitro evaluation techniques
Dong MEI ; Long-tai YOU ; Huan ZHANG ; Xiao-ling WANG
Acta Pharmaceutica Sinica 2024;59(6):1847-1853
Oral iron is a commonly used preparation for the treatment of iron deficiency and iron-deficiency anemia in children, but its undesirable taste has become a major factor affecting clinical adherence to the medication. No studies have been conducted to evaluate the palatability of oral iron supplements. Thirteen representative oral iron supplements were selected to evaluate the palatability of oral iron supplements from different perspectives of
3.Stability and taste evaluation of two kinds of oral solvent after extemporaneous compounding
Wei-wei GUAN ; Min HUANG ; Huan ZHANG ; Long-tai YOU ; Li-ping FENG ; Min YANG ; Dong MEI ; Xiao-ling WANG
Acta Pharmaceutica Sinica 2023;58(11):3198-3203
A flavoring agent and a suspension agent were prepared for extemporaneous compounding. The stability of the two agents before and after drug loading was investigated, and the taste of the suspension after extemporaneous compounding was evaluated by electronic tongue technology. The two agents remained stable under the conditions of influence factor test, accelerated test and long-term test. The appearance properties of the two agents did not change. The relative density of the flavoring agent was maintained at 1.053-1.075, and the pH was stable at 4.2-4.5. The relative density of the suspension agent was maintained at 0.999-1.022, and the pH was stable at 4.0-4.5. Seven kinds of drugs, including warfarin sodium tablets and spironolactone tablets, were mixed with these two oral solvents, and the content uniformity and stability were detected respectively. The results showed that the preparations could be evenly dispersed and the physical and chemical properties were stable. The results of taste evaluation showed that in captopril group and chloral hydrate group, the flavoring agent had the best effect on taste correction. In warfarin sodium group, rifampicin group, spironolactone group, vitamin B1 group and vitamin B2 group, the blending agents had the best effect on taste correction.
4.Mortality trend of chronic respiratory diseases in China, 1990-2019.
Zheng LONG ; Wei LIU ; Jin Lei QI ; Yun Ning LIU ; Jiang Mei LIU ; Jin Ling YOU ; Lin LIN ; Li Jun WANG ; Mai Geng ZHOU ; Peng YIN
Chinese Journal of Epidemiology 2022;43(1):14-21
Objective: To analyze mortality and its trend of chronic respiratory diseases (CRD) in China from 1990 to 2019. Methods: Based on the provincial results of China from the 2019 Global Burden of Disease (GBD) study, the average annual percent change (AAPC) of standardized mortality rates of different CRDs were analyzed by using Joinpoint 4.8.0.1, and the age-standardized mortality rate of CRD was calculated by using the GBD 2019 world standard population. Based on the comparative risk assessment theory of GBD, the attributable deaths due to 12 CRD risk factors were estimated, including smoking, indoor air pollution, occupational gas exposure, particulates and smog exposure, environmental particulate pollution, low temperature, passive smoking, ozone pollution, occupational exposure to silica, occupational asthma, high body mass index, high temperature and occupational exposure to asbestos. Results: From 1990 to 2019, the number of deaths and standardized mortality of chronic obstructive pulmonary disease (COPD) showed a downward trend (P<0.001). The number of COPD deaths decreased from 1 244 000 (912 000 - 1 395 000) in 1990 to 1 037 000 (889 000 - 1 266 000) in 2019. AAPC=-0.9% (95%CI: -1.5% - -0.3%), P<0.001; The standardized mortality rate decreased from 217.9/100 000 (163.3/100 000 - 242.0/100 000) in 1990 to 65.2/100 000 (55.5/100 000 - 80.1/100 000) in 2019. AAPC= -4.2% (95%CI:-5.2% - -3.2%), P<0.001. The number of deaths from asthma decreased from 40 000 (30 000 - 58 000) in 1990 to 25 000 (20 000 - 31 000) in 2019. AAPC=-2.0% (95%CI: -2.6% - -1.4%), P<0.001; The standardized mortality rate of asthma decreased from 6.4/100 000 (4.7/100 000 - 9.5/100 000) in 1990 to 1.5/100 000 (1.2/100 000 - 1.9/100 000) in 2019. AAPC=-5.1% (95%CI: -5.8% - -4.4%), P<0.001. The number of pneumoconiosis deaths decreased from 11 000 (8 000 - 14 000) in 1990 to 10 000 (8 000 - 14 000) in 2019, AAPC=-0.2%(95%CI:-0.4% - 0.1%), P=0.200; The standardized mortality rate of pneumoconiosis decreased from 1.4/100 000 (1.0/100 000 - 1.7/100 000) in 1990 to 0.5/100 000 (0.4/100 000 - 0.7/100 000) in 2019. AAPC=-3.1% (95%CI: -3.4% - -2.8%), P<0.001. The number of deaths from pulmonary interstitial diseases and pulmonary sarcoidosis increased from 3 000 (3 000 - 6 000) in 1990 to 8 000 (6 000 - 10 000) in 2019, AAPC=3.5% (95%CI: 2.7% - 4.2%), P<0.001; The corresponding standardized mortality rate changed little from 1990 to 2019, and AAPC was not statistically significant.The age-standardized mortality rates of different CRDs were higher in men than those in women. In 1990 and 2019, the mortality rates of COPD, asthma, pneumoconiosis and interstitial pulmonary disease and pulmonary sarcoidosis increased with age. In 2019, the population attributable fractions (PAFs) for smoking, environmental particulate pollution, occupational gas exposure, particulate and smog exposure, low temperature exposure and passive smoking were 71.1% (68.0% - 74.3%), 24.7% (20.1% - 30.0%), 19.3% (13.0% - 25.4%), 15.7% (13.6% - 18.3%) and 8.8% (4.5% - 13.1%) respectively in men, and the PAFs for environmental particulate pollution, smoking, low temperature exposure, occupational gas exposure, particulate and smog exposure, and passive smoking were 24.1% (19.6% - 29.3%), 21.9% (18.7% - 25.2%), 16.4% (14.0% - 19.2%), 15.6% (10.2% - 21.1%) and 14.7% (7.9% - 21.3%) respectively in women. Conclusions: During 1990-2019, the overall death level of CRD decreased significantly in China, but it is still at high level in the world. Active prevention and control measures should be taken to reduce the death level caused by CRD.
Asthma
;
China/epidemiology*
;
Female
;
Global Burden of Disease
;
Humans
;
Male
;
Mortality
;
Pulmonary Disease, Chronic Obstructive
;
Quality-Adjusted Life Years
5.Expert consensus on the diagnosis and treatment of severe and critical coronavirus disease 2019.
You SHANG ; Jianfeng WU ; Jinglun LIU ; Yun LONG ; Jianfeng XIE ; Dong ZHANG ; Bo HU ; Yuan ZONG ; Xuelian LIAO ; Xiuling SHANG ; Renyu DING ; Kai KANG ; Jiao LIU ; Aijun PAN ; Yonghao XU ; Changsong WANG ; Qianghong XU ; Xijing ZHANG ; Jicheng ZHANG ; Ling LIU ; Jiancheng ZHANG ; Yi YANG ; Kaijiang YU ; Xiangdong GUAN ; Dechang CHEN
Chinese Medical Journal 2022;135(16):1913-1916
Humans
;
COVID-19
;
Consensus
;
SARS-CoV-2
;
China
6.Effect of acupuncture on pain and cerebral hemodynamics in patients with migraine: a randomized controlled trial.
Kai-Feng DENG ; Xue LI ; Hui-Ling LU ; Heng NING ; Xiao-Hua YOU ; Ying ZHU ; Ri-Lan CHEN ; Zi-Long LIAO
Chinese Acupuncture & Moxibustion 2021;41(2):115-120
OBJECTIVE:
To observe the therapeutic effect between acupuncture combined with medication and simple medication on migraine and cerebral hemodynamics.
METHODS:
A total of 120 patients with migraine were randomized into an acupuncture plus medication group (60 cases, 3 cases dropped off) and a medication group (60 cases, 6 cases dropped off). In the medication group, flunarizine hydrochloride capsule was given orally before sleep, 10 mg a day. On the basis of the treatment in the medication group, acupuncture was applied at Sizhukong (TE 23), Shuaigu (GB 8), Taiyang (EX-HN 5), Fengchi (GB 20) and etc. in the acupuncture plus medication group, 30 min each time, once a day. Treatment for 4 weeks was required in both groups. Before and after treatment, the visual analogue scale (VAS) score, indexes of cerebral hemodynamic [blood flow velocity of anterior cerebral artery (ACA), middle cerebral artery (MCA), posterior cerebral artery (PCA), vertebral artery (VA) and basilar artery (BA)] and total TCM syndrome score were observed, and the clinical therapeutic effect and the incidence of the adverse events were evaluated in both groups.
RESULTS:
Compared before treatment, the VAS scores, the blood flow velocity of ACA, MCA, PCA, VA, BA and the total TCM syndrome scores were decreased in both groups (
CONCLUSION
Acupuncture combined with flunarizine hydrochloride capsule can effectively relieve the pain in patients with migraine, reduce the cerebral blood flow velocity, the efficacy is superior to simple flunarizine hydrochloride capsule.
Acupuncture Points
;
Acupuncture Therapy
;
Hemodynamics
;
Humans
;
Migraine Disorders/therapy*
;
Pain
;
Treatment Outcome
7.Effects of Xueshuantong Injection on thrombosis formation and blood flow in rats.
Lei LIU ; Shi-Lan DING ; Ying CHEN ; Qiong-Ling ZHANG ; Qian ZHANG ; Zheng-Xiao SUN ; Fu-Long LIAO ; Yun YOU
China Journal of Chinese Materia Medica 2020;45(10):2446-2453
The aim of this paper was to explore the effect of Xueshuantong Injection(freeze-dried powder,XST) on κ-carrageenan-induced thrombosis and blood flow from the aspects of interactions among blood flow,vascular endothelium and platelets. Fifty male Sprague-Dawley rats(190-200 g) were randomized into five groups: control group, model group, heparin sodium(1 000 U·kg~(-1)) group, low-dose and high-dose(50, 150 mg·kg~(-1)) XST groups. Rats were intraperitoneally injected with corresponding drugs and normal saline(normal control and model groups) for 10 days. One hour after drugs were administered intraperitoneally on the 7 th day, each rat was injected with κ-carrageenan(Type Ⅰ, 1 mg·kg~(-1)) which was dissolved in physiological saline by intravenous administration in the tail to establish tail thrombus model. The lengths of black tails of the rats were measured at 2, 6, 24 and 48 h after modeling. Vevo®2100 small animal ultrasound imaging system was used to detect the internal diameter of rat common carotid artery, blood flow velocity and heart rate, and then the blood flow and shear rate were calculated. Meanwhile, the microcirculatory blood flow perfusion in the thigh surface and tail of rats were detected by laser speckle blood flow imaging system. Platelet aggregometry was used to detect the max platelet aggregation rate in rats. Pathological changes in tail were observed through hematoxylin-eosin staining, and Western blot was used to detect the protein content of platelet piezo1. According to the results, XST could inhibit the rat tail arterial thrombosis and significantly reduce the length of black tail(P<0.05). The blood flow of common carotid artery in XST low dose group was significantly higher than that in the model group(P<0.05). XST high dose group could significantly increase the microcirculatory blood flow perfusion of the tail in rats as compared with the model group(P<0.05). XST high dose group could significantly inhibit platelet aggregation rate(P<0.05) and XST low dose group could significantly inhibit platelet piezo1 protein expression(P<0.01). In summary, XST could play an effect in fighting against thrombosis induced by κ-carrageenan in rats, which may be related to significantly inhibiting platelet aggregation, improving body's blood flow state, maintaining normal hemodynamic environment and affecting mechanical ion channel protein piezo1.
Animals
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Drugs, Chinese Herbal
;
Male
;
Microcirculation
;
Rats
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Rats, Sprague-Dawley
;
Thrombosis
8.Efficacy of cardiopulmonary ultrasound in guiding volume therapy in patients with sepsis-induced myocardial injury
Cong HE ; Shan REN ; You FU ; Limin SHEN ; Heling ZHAO ; Ling LONG ; Xianlei WANG
Chinese Journal of Anesthesiology 2019;39(3):373-376
Objective To evaluate the effect of cardiopulmonary ultrasound in guiding volume ther-apy in the patients with sepsis-induced myocardial injury. Methods Thirty-eight patients of both sexes with septic myocardial injury, aged 28-64 yr, weighing 60-90 kg, received anti-infection, respiratory and circulatory comprehensive treatment. The patients were divided into group Ⅰ ( n=19) and group Ⅱ(n=19) by using a random number table method. Central venous pressure (CVP) was used to guide vol-ume therapy, and fluid replacement was carried out using the CVP 2-5 principle in groupⅠ. Cardiopulmo-nary ultrasound was used to guide volume therapy in group Ⅱ. Blood samples were taken before volume therapy and on 1, 3 and 5 days after volume therapy to determine the concentrations of N-terminal pro-B-type natriuretic peptide, cardiac troponin I and creatine kinase isoenzyme MB in serum. The CVP, positive fluid balance, lactic acid, central venous oxygen saturation and urine volume were recorded at 6, 24, 48 and 72 h after volume therapy. Left ventricular ejection fraction was recorded at 1, 3 and 5 days after vol-ume therapy. The length of intensive care unit stay and 28-day fatality were recorded. Results Compared with groupⅠ, the CVP and fluid positive balance were significantly decreased at each time point after vol-ume therapy, the concentration of serum N-terminal pro-B-type natriuretic peptide was decreased at 5 days after volume therapy ( P<0. 05) , and no significant difference was found in concentrations of cardiac tropo-nin I and creatine kinase isoenzyme MB in serum, lactic acid, central venous oxygen saturation and urine volume, left ventricular ejection fraction, length of intensive care unit stay or 28-day fatality rate at each time point in groupⅡ( P>0. 05) . Conclusion Cardiopulmonary ultrasound can reduce the volume of liq-uid infused, avoid fluid overload and avoid accentuating myocardial injury when used to guide volume thera-py in the patients with sepsis-induced myocardial injury.
9.Study on Ambi-extracting and Inclusion Process of Volatile Oil from Chuanxiong Rhizoma and Angelicae Sinensis Radix
Mei-Ling WANG ; Jian NI ; Lin GUO ; Gong-Sen CHEN ; Ying-Chao YU ; Long-Tai YOU ; Hai-Ying ZHANG ; Xing-Bin YIN ; Chang-Hai QU
Chinese Journal of Information on Traditional Chinese Medicine 2018;25(11):68-72
Objective To optimize ambi-extracting and inclusion process of volatile oil from Chuanxiong Rhizoma and Angelicae Sinensis Radix. Methods With yield ratio of volatile oil and ferulic acid content in water extract as evaluation indexes, single factor experiments were used to study the extraction process. With the inclusion rate of volatile oil and yield of inclusion as evaluation indexes, saturated aqueous solution was used to L9(34) orthogonal experiments to reach optimum inclusion process. Results The optimum extraction process of Chuanxiong Rhizoma and Angelicae Sinensis Radix was extracted for 8 hours with 8 folds the amount of water, and without soaking. The validation experiments of extraction of volatile oil and ferulic acid content in water extract were 1.23 mL and 0.387 9 mg/g. The optimum conditions of inclusion process were as follows: volatile oil (mL): β-CD (g) was 1:8;inclusion temperature was 40 ℃; inclusion time was 3 hours. The validation experiments of inclusion rate of volatile oil and yield of inclusion were 74.89% and 72.81%. Conclusion Optimum ambi-extracting and inclusion process of volatile oil from Chuanxiong Rhizoma and Angelicae Sinensis Radix are feasible and stable, witch can provide certain supporting data for preparation production.
10.Observations on the Therapeutic Effect of Kinetic Rectification Acupuncture on Acute Facial Neuritis
Zu-Hong WANG ; Chun-Yan GUO ; Pei-Dong HUANG ; You-Ling LONG ; Rong YI ; Xiao-Rong DUAN ; Li LI
Shanghai Journal of Acupuncture and Moxibustion 2018;37(2):205-208
Objective To investigate the clinical efficacy of kinetic rectification acupuncture in treating acute facial neuritis. Method Sixty patients with acute facial neuritis were randomized to observation and control groups. The observation group received kinetic rectification acupuncture and the control group, conventional acupuncture alone. Acupuncture was given five times a week, five times as one course. The therapeutic effects were evaluated after three courses of treatment. Result The total efficacy rate was 93.3% in the observation group and 73.3% in the control group; there was a statistically significant difference between the two groups (P<0.05). The latencies and amplitudes of the frontal muscle, orbicularis oculi muscle and quadrate muscle of upper lip improved in the two groups after treatment and had statistically significant pre-/post-treatment differences (P < 0.01). There were statistically significant differences in the pre-/post-treatment difference values of the latencies and amplitudes of the frontal muscle and orbicularis oculi muscle (P<0.01) and no statistically significant difference in the pre-/post-treatment difference values of the latency and amplitude of the quadrate muscle of upper lip (P>0.05) between the two groups. Conclusion Kinetic rectification acupuncture has a marked therapeutic effect on acute facial neuritis. This study provides a particular therapeutic method for clinical practice.

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