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.Fatty Acid Binding Protein 5 (FABP5) Promotes Aggressiveness of Gastric Cancer Through Modulation of Tumor Immunity
Mei-qing QIU ; Hui-jun WANG ; Ya-fei JU ; Li SUN ; Zhen LIU ; Tao WANG ; Shi-feng KAN ; Zhen YANG ; Ya-yun CUI ; You-qiang KE ; Hong-min HE ; Shu ZHANG
Journal of Gastric Cancer 2023;23(2):340-354
Purpose:
Gastric cancer (GC) is the second most lethal cancer globally and is associated with poor prognosis. Fatty acid-binding proteins (FABPs) can regulate biological properties of carcinoma cells. FABP5 is overexpressed in many types of cancers; however, the role and mechanisms of action of FABP5 in GC remain unclear. In this study, we aimed to evaluate the clinical and biological functions of FABP5 in GC.
Materials and Methods:
We assessed FABP5 expression using immunohistochemical analysis in 79 patients with GC and evaluated its biological functions following in vitro and in vivo ectopic expression. FABP5 targets relevant to GC progression were determined using RNA sequencing (RNA-seq).
Results:
Elevated FABP5 expression was closely associated with poor outcomes, and ectopic expression of FABP5 promoted proliferation, invasion, migration, and carcinogenicity of GC cells, thus suggesting its potential tumor-promoting role in GC. Additionally, RNA-seq analysis indicated that FABP5 activates immune-related pathways, including cytokinecytokine receptor interaction pathways, interleukin-17 signaling, and tumor necrosis factor signaling, suggesting an important rationale for the possible development of therapies that combine FABP5-targeted drugs with immunotherapeutics.
Conclusions
These findings highlight the biological mechanisms and clinical implications of FABP5 in GC and suggest its potential as an adverse prognostic factor and/or therapeutic target.
3.Analysis of mental health status of Hainan residents
HONG Jian-he ; GAO Li ; WANG Xiao-dan ; DU Zhen-you ; LI Zhi-yun ; ZUO Qiu-hui ; WENG Xin-mei
China Tropical Medicine 2023;23(3):267-
Abstract: Objective To understand the mental health literacy level of residents in Hainan Province, and to provide evidence for promoting mental health promotion and improving the mental health literacy level of residents. Methods Multi-stage random sampling method was used to investigate The National Mental Health Literacy Questionnaire among 6 895 residents in 12 districts including Changjiang, Chengmai, Dongfang, Ledong, Lingshui, Haikou Longhua District, Haikou Meilan District, Qionghai, Sanya, Tunchang, Wenchang and Wuzhishan. Results In the survey 6 895 residents in 12 counties, cities, districts of Hainan Province, 365 of them reached the standard of mental health with the standard rate of 5.3%. The mental health knowledge score was (54.00±17.02) with the standard rate of 7.7%; the self-assessment score was (26.11±3.99) with the standard rate of 75.9%; the score of mental health skill was (26.22±7.25) with the standard rate of 44.8%. The mental health literacy level of medical workers was the highest, with the standard rate of 13.5% (112/830), and the mental health literacy level of farmers was the lowest, with the standard rate of 0.8% (13/1 647). The results of unconditional Logistic regression showed that the factors entering the regression model included educational background (OR=2.268), personal monthly income (OR=1.129), gender (OR=1.302), household registration (OR=0.776), and whether they had participated in mental health related courses OR training (OR=0.511). The higher the educational background and personal monthly income, the higher the psychological quality. The mental health of women was higher than that of men, and that of urban was higher than that of rural, and those who had participated in mental health related courses were higher than those who had not. Conclusions The mental health literacy level of Hainan residents is at a low level, and the influencing factors are multifaceted. It is suggested to strengthen the mental health monitoring comprehensively and carry out rich health education service mode for different groups.
4.Effect of Suanzaoren Tang on Energy Metabolism of Myocardial Mitochondria in Aged Rats with Chronic Rapid Eye Movement Sleep Deprivation
Xin LIU ; Ping WANG ; Li DING ; Shun-bo ZHANG ; Kun SHI ; Min-min YANG ; Qiu-yun YOU
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(16):40-46
Objective:To study the mechanism of Suanzaoren Tang in regulating the energy metabolism of myocardial mitochondria in aged rats with chronic rapid eye movement (REM) sleep deprivation through the sirtuin 3 (SIRT3)/superoxide dismutase2 (SOD2) signaling pathway. Method:Fifty male Wistar rats were randomly divided into the control group, model group, estazolam group (0.18 mg·kg-1·d-1), and low- (6.48 g·kg-1·d-1) and high-dose (12.96 g·kg-1·d-1) Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of
5.Effect of Suanzaoren Tang on Mitochondrial Damage and Apoptosis in Hypothalamus of Elderly Rats with Chronic Rapid Eye Movement Sleep Deprivation
Min-min YANG ; Xin LIU ; Qiu-yun YOU ; Shun-bo ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(16):47-52
Objective:To investigate the effect of Suanzaoren Tang on mitochondria-mediated neuronal apoptosis. Method:Fifty male Wistar rats were randomly divided into the control group, model group, estazolam group (0.18 mg·kg-1·d-1), and low- (6.48 g·kg-1·d-1) and high-dose (12.96 g·kg-1·d-1) Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of
6.Effect of Suanzaoren Tang on Energy Metabolism of Liver Mitochondria in Aged Rats with Chronic Rapid Eye Movement Sleep Deprivation and Its Mechanism
Xin LIU ; Qiu-yun YOU ; Ping WANG ; Li DING ; Fu-nan NING ; Shun-bo ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(16):53-58
Objective:To study the effect of Suanzaoren Tang on energy metabolism of liver mitochondria in aged rats with chronic rapid eye movement (REM) sleep deprivation. Method:Fifty male Wistar rats were randomly divided into the control group, model group, estazolam group (0.18 mg·kg-1·d-1), and low- (6.48 g·kg-1·d-1) and high-dose (12.96 g·kg-1·d-1) Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of
7.Suanzaoren Tang Treats Insomnia by Affecting Brain, Heart, and Liver Functions: A Review
Zi-heng LI ; Xin LIU ; Ping WANG ; Qiu-yun YOU ; Li CHENG ; Li DING ; Shun-bo ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(16):59-66
Sleep has been widely concerned by the medical field all over the world. Sleep deprivation can cause damage to organs of the human body, which is related to the occurrence of a variety of diseases. Besides, the pathological change in different organs of the human body is also a key factor that causes or aggravates insomnia. When treating insomnia and its complications, traditional Chinese medicine (TCM) focuses on the homology of the brain and heart, and insomnia is mainly treated from the five internal organs, especially the heart and liver. Sleep duration and structure change with age. The prevalence of insomnia is higher in older individuals susceptible to complications than in the younger population. In TCM, insomnia of blood deficiency and Yin deficiency is common among the elderly. Suanzaoren Tang is a classic prescription for nourishing blood and calming the mind and it is critical in the treatment of "sleeplessness due to consumptive disease and dysphoria", with the effects of nourishing liver blood to calm the mind and clearing internal heat to relieve dysphoria. It has good efficacy on the insomnia of the elderly caused by deficiency of Qi and blood and abnormal operation of nutrient Qi and defense Qi. Furthermore, it also shows a certain therapeutic effect on insomnia combined with cardiovascular and cerebrovascular diseases. The present study revealed the damage to the brain, heart, and liver caused by sleep deprivation and the effect of Suanzaoren Tang on the brain, heart, and liver, and clarified the facts that Suanzaoren Tang inhibited the damage to organs caused by sleep deprivation and regulated energy metabolism, thereby exploring the sedative and hypnotic mechanism of Suanzaoren Tang to provide new ideas for Suanzaoren Tang in the treatment of sleep disorders and other diseases.
8. Effect of Shenghuitang on Learning Memory, Levels of IL-6, TNF-α and COX-2 in Hippocampus of Chronic Sleep Deprivation Mice
Chao YIN ; Qiu-yun YOU ; Mei-ya ZHANG ; Li DING
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(1):96-100
Objective: To observe the effect of Shenghuitang on learning and memory and expressions of interleukin-6(IL-6), tumor necrosis factor-α(TNF-α) and cyclooxygenase-2(COX-2) in hippocampus of chronic sleep deprived mice, in order to explore the possible mechanism of Shenghuitang in improving learning and memory ability. Method: Mice were randomly divided into sleep deprivation group, blank group, melatonin group(7.8×10-4 g·kg-1·d-1), high, middle and low-dose Shenghuitang groups(54,27,13.5 g·kg-1·d-1). The model of chronic sleep deprivation in mice was established using the "multi-platform water environment method". 28 d sleep deprivation and intragastric administration were provided. Morris water maze was used to detect the learning and memory ability of mice in each group. Real time-PCR was used to detect mRNA expressions of IL-6, TNF-α and COX-2 in the hippocampus of each group. Result: The results of Morris water maze test showed that compared with the blank group, the total time spent on finding the platform and the total swimming distance of the model group were significantly prolonged (P<0.01), while the number of crossing platforms and the target quadrant were significantly reduced (P<0.01). The time for the original platform was significantly increased (P<0.01). Compared with the model group, the total time spent on finding the platform and the total swimming distance decreased significantly in each drug-administered group (P<0.05,P<0.01) reduced, whereas the number of times for crossing the platform and the target quadrant increased significantly (P<0.05,P<0.01). The time for the first arrival of the original platform was significantly reduced (P<0.05,P<0.01). The results of RT-PCR showed that mRNA expressions of IL-6, TNF-α, and COX-2 were increased in the model group compared with the blank group. Compared with the model group, mRNA expressions of IL-6, TNF-α, and COX-2 were decreased in the treated group. COX-2 mRNA expression was down-regulated. Conclusion: Shenghuitang may improve the learning and memory ability of mice by decreasing mRNA expressions of IL-6, TNF-α and COX-2 in hippocampus.
9. Effect of Shenghuitang on Bmal1 in Hypothalamus and IL-6 and TNF-α in Hippocampus of APP/PS1 Double Transgenic Dementia Model Mice
Mei-ya ZHANG ; Ping WANG ; Qiu-yun YOU ; Li DING
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(20):7-12
Objective:To investigate the effect of Shenghuitang on learning and memory, biological clock gene[brain and muscle arnt-like 1 (Bmal1)] in hypothalamus and interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in hippocampus of APP/PS1 double transgenic dementia model mice, in order to explore the possible mechanism of Shenghuitang to improve learning and memory and sleep disorders. Method:The experimental mice were randomly divided into model group, blank control group, melatonin group, high-dose Shenghuitang group and low-dose Shenghuitang group. Autonomic activity analysis system was used to detect the autonomic activities of mice in each group. Morris water maze was used to detect the learning ability and spatial memory ability of each group. quantitative real-time fluorescence polymerase chain reaction(Real-time PCR) was used to detect the expression of Bmal1 mRNA in the hypothalamic area of mice. Western blot was used to detect the expression of Bmal1 protein in each group. The content of inflammatory factors IL-6 and TNF-α in hippocampus of mice was detected by enzyme-linked immunosorbent assay(ELISA). The correlation between inflammatory factors IL-6, TNF-α and Bmal1 gene was analyzed by pearson analysis. Result:The results of voluntary activities showed that compared with the control group, the number of activities and activity distance of the model group were significantly decreased (P<0.01). Compared with the model group, the number of activities and activity distance of the mice in each drug group increased significantly (P<0.05, P<0.01), there was no significant difference in the low dose group of Shenghuitang. Morris water maze results showed that compared with the control group, the platform latency and swimming total distance were significantly prolonged in the model group (P<0.01), and the number of crossing platforms and target quadrant time was significantly reduced (P<0.01). The original platform time increased significantly (P<0.05). Compared with the model group, the platform latency and total swimming distance were significantly decreased in each group (P<0.05, P<0.01), and the number of crossing platforms and target quadrant time increased significantly(P<0.05, P<0.01), the time to the original platform was significantly reduced (P<0.05, P<0.01). Real-time PCR results showed that the expression of Bmal1 mRNA was up-regulated in the model group compared with the control group. Compared with the model group, the mRNA expression of Bmal1 gene was down-regulated in each group. Western blot results showed that compared with the control group, the expression of Bmal1 protein in the model group was significantly increased (P<0.01). Compared with the model group, Bmal1 protein expression was significantly decreased in each group(P<0.01). The results of ELISA showed that the levels of IL-6 and TNF-α in the model group were significantly higher than those in the control group (P<0.01). Compared with the model group, the levels of IL-6 and TNF-α in the drug group were significantly lower(P<0.01). Pearson analysis showed that IL-6, TNF-α and Bmal1 were correlated and negatively correlated. Conclusion:Shenghuitang may reduce the levels of inflammatory factors IL-6 and TNF-α in hippocampus by up-regulating the expression of Bmal1 gene in hypothalamic region, thus improving Alzheimer' s disease(AD) and circadian rhythm disorders.
10.Preliminary study on the risk of macrosomia using Bayesian discriminant analysis based on prenatal records
Liu-yu WU ; Jing-you LAN ; Dan-dan HUANG ; Xiao-qiang QIU ; Mei-liang LIU ; Qiu-li LIANG ; Di ZHANG ; Xiao-yun ZENG
Chinese Journal of Disease Control & Prevention 2019;23(11):1338-1341,1347
Objective To explore the clinical effect of Bayesian discriminant analysis in predicting the risk of macrosomia. Methods 169 fetal macrosomia and 169 non-macrosomia were enrolled in a 1:1 matched case-control study. Conditional Logistic regression was used to select the discriminant indexes,and the discriminant indexes were put into the Bayesian discriminant model to obtain the Bayesian discriminant function. The discriminant function was the retrospectively examined and externally tested. Results The results of conditional Logistic regression model indicated that mother's height, early pregnancy body mass index (BMI), gestational diabetes, gestational weeks, the height of uterine and abdominal circumference were associated with the birth of fetal macrosomia. The Bayesian discriminant function were established: Fetal macrosomia:y1=-27.802+8.420×Mother's height+8.719×early pregnancy BMI+10.485×gestational weeks+3.375×gestational diabetes+2.862×height of uterine and abdominal circumference; Non-macrosomia y2=-17.477+7.161×Mother's height+7.217×early pregnancy BMI+7.862×gestational weeks+2.036×gestational diabetes-0.085×height of uterine and abdominal circumference. Wilks′ Lambda λ=0.489, P<0.001, the Bayesian discriminant function was statistically significant. The internal and external conformity rates of the Bayesian discriminant model were all more than 80%. Conclutions The birth of fetal macrosomia is related to many factors. The Bayesian discriminant model in the present study is valuable to discriminate macrosomia and provide an objective reference for more accurate identification of macrosomia in the future.

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