1.Anemoside B4 inhibits SARS-CoV-2 replication in vitro and in vivo.
Mingyue XIAO ; Ronghua LUO ; Qinghua LIANG ; Honglv JIANG ; Yanli LIU ; Guoqiang XU ; Hongwei GAO ; Yongtang ZHENG ; Qiongming XU ; Shilin YANG
Chinese Herbal Medicines 2024;16(1):106-112
OBJECTIVE:
Anemoside B4 (AB4), the most abundant triterpenoidal saponin isolated from Pulsatilla chinensis, inhibited influenza virus FM1 or Klebsiella pneumoniae-induced pneumonia. However, the anti-SARS-CoV-2 effect of AB4 has not been unraveled. Therefore, this study aimed to determine the antiviral activity and potential mechanism of AB4 in inhibiting human coronavirus SARS-CoV-2 in vivo and in vitro.
METHODS:
The cytotoxicity of AB4 was evaluated using the Cell Counting Kit-8 (CCK8) assay. SARS-CoV-2 infected HEK293T, HPAEpiC, and Vero E6 cells were used for in vitro assays. The antiviral effect of AB4 in vivo was evaluated by SARS-CoV-2-infected hACE2-IRES-luc transgenic mouse model. Furthermore, label-free quantitative proteomics and bioinformatic analysis were performed to explore the potential antiviral mechanism of action of AB4. Type I IFN signaling-associated proteins were assessed using Western blotting or immumohistochemical staining.
RESULTS:
The data showed that AB4 reduced the propagation of SARS-CoV-2 along with the decreased Nucleocapsid protein (N), Spike protein (S), and 3C-like protease (3CLpro) in HEK293T cells. In vivo antiviral activity data revealed that AB4 inhibited viral replication and relieved pneumonia in a SARS-CoV-2 infected mouse model. We further disclosed that the antiviral activity of AB4 was associated with the enhanced interferon (IFN)-β response via the activation of retinoic acid-inducible gene I (RIG-1) like receptor (RLP) pathways. Additionally, label-free quantitative proteomic analyses discovered that 17 proteins were significantly altered by AB4 in the SARS-CoV-2 coronavirus infections cells. These proteins mainly clustered in RNA metabolism.
CONCLUSION
Our results indicated that AB4 inhibited SARS-CoV-2 replication through the RLR pathways and moderated the RNA metabolism, suggesting that it would be a potential lead compound for the development of anti-SARS-CoV-2 drugs.
2.Analysis of depressive symptoms and influencing factors among middle and high school students from 2018 to 2021 in Zhejiang Province
GU Fang, YANG Ying, ZHENG Weijun, MENG Jia, LI Juanjuan, SHEN Yu, GAO Lei, ZOU Yan, ZHANG Ronghua
Chinese Journal of School Health 2024;45(4):520-524
Objective:
To investigate the prevalence and associated factors of depressive symptoms among middle and high school students in Zhejiang Province, so as to provide scientific basis for the implementation of depressive intervention.
Methods:
Based on the health status and associated factors of middle and high school students in the project "Monitoring of Common Diseases and Health Influencing Factors of Students" during 2018 to 2021, a total of 73 309 students including middle school, ordinary high school and vocational high school surveyed in 11 cities of Zhejiang Province were selected by multi stage stratified cluster random sampling method. From 2018 to 2021, there were 6 008, 21 917, 23 712 and 21 672 students, respectively. The Chi square test and Logistic regression model were used to analyze the influencing factors of depressive symptoms in middle and high school students.
Results:
From 2018 to 2021, depressive symptoms detection rate of middle school students was 14.8%, with higher rate in girls (17.1%) than in boys (12.7%), higher rate in high school (17.1% in ordinary high school, 17.6% in vocational high school) than middle school (12.5%)( χ 2=278.77, 327.22, P <0.05). Univariate analysis showed that there were statistically significant differences in depressive symptoms detection rate among middle school students with different years (2018: 16.7%,2019: 17.9% , 2020: 13.1%, 2021: 13.0%), residence (yes: 16.3%, no:13.5%), body mass index classification (not overweight or obesity: 14.8%, overweight: 14.2%, Obesity: 15.7%), weekly exercise days (0-2 d: 17.1%, 3-5 d: 12.5%, 6-7 d: 13.1%) and bullying (yes: 35.5%, no: 10.7%) ( χ 2=293.40, 118.35, 7.83, 287.24, 4 978.84, P <0.05). Multivariate Logistic regression analysis showed that female students, ordinary high schools, vocational high schools, obesity, school bullying were positively correlated with depression ( OR =1.65, 1.70, 1.60, 1.12, 5.21), exercise 3 to 5 days per week, exercise 6 to 7 days per week were negatively correlated with depression ( OR=0.77, 0.81, P <0.01).
Conclusions
Depressive symptoms among middle and high school students in Zhejiang Province are prominent. Strengthening mental health education for students and providing attention and support from families, schools, and society are essential steps to reduce the occurrence of depressive symptoms among these students.
3.Researchon the training model of innovative talents in traditional Chinese medicine with the integration of science and education in local high-level universities
Lanwen GAO ; Ye GAO ; Ronghua ZHANG ; Li YANG ; Huan WANG ; Xiaoyun LI ; Lingyu LI ; Yu CAI
Modern Hospital 2024;24(1):127-129
Traditional Chinese Medicine has a long history and plays a decisive role in the fields of modern medicine and pharmacy.It is an important part of our country's traditional medicine.With the progress of the times,people are paying more and more attention to the innovation and development of traditional Chinese medicine.However,the current traditional Chinese medicine talents trained by major universities cannot meet the needs and requirements of society.This is closely related to the current talent training model of universities.Local high-level universities have unique advantages and potential in cultivating inno-vative talents in traditional Chinese medicine.They can incorporate traditional Chinese medicine culture with local characteristics into the teaching content and practical links of training traditional Chinese medicine talents,and build innovative traditional Chi-nese medicine talents integrating science and education.The training model is more conducive to cultivating top innovative talents that meet the needs of society and and the development of traditional Chinese medicine.This article analyzes and discusses how local high-level universities can develop innovative talent training models that suit the needs of traditional Chinese medicine by combining local characteristics and disciplinary advantages,so as to provide useful reference and inspiration for local high-level u-niversities in cultivating talents in traditional Chinese medicine.
4.Efficacy and Mechanism of Lutongning Granules in Treatment of Trigeminal Neuralgia Induced by Injection of Talc into Infraorbital Foramen of Model Rats Based on P2X7R-mediated Neuroinflammation
Qiyue SUN ; Shuran LI ; Shuangrong GAO ; Shanshan GUO ; Zihan GENG ; Lei BAO ; Ronghua ZHAO ; Jingsheng ZHANG ; Bo PANG ; Yingli XU ; Yu ZHANG ; Shan CAO ; Yaxin WANG ; Xiaolan CUI ; Bing HAN ; Jing SUN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):56-63
ObjectiveTo evaluate the effectiveness of Lutongning granules in the treatment of trigeminal neuralgia in animal models and study its mechanism of action, so as to provide laboratory data support for the clinical application of Lutongning granules and precise treatment. MethodMale SD rats were randomly divided into normal group, model group, carbamazepine group (0.06 g·kg-1·d-1), high-dose Lutongning group (2.70 g·kg-1·d-1), and low-dose Lutongning group (1.35 g·kg-1·d-1) according to the stratified basic mechanical pain thresholds, with 10 rats in each group. A trigeminal neuralgia model of rats was prepared by injecting 30% talc suspension into the infraorbital foramen area of the rat. The drug groups were administered 10 mL·kg-1 of drugs by gavage after 2 h of modeling. The normal group and the model group were administered distilled water by gavage under the same conditions once a day for 10 consecutive days. Von Frey brushes were used to determine the mechanical pain threshold of rats. A fully automated blood and body fluid analyzer was employed to detect the blood routine of rats. Hematoxylin and eosin (HE) staining was utilized to detect the pathological changes in the trigeminal ganglion and medulla oblongata tissue. Transmission electron microscopy was used to scan the ultrastructure of the medulla oblongata tissue. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors interleukin (IL)-1, IL-6, IL-8, tumor necrosis factor (TNF)-α, neuropeptide substance P, and β-endorphins (β-EP) in the serum of rats, and Western blot was used to detect the protein expression levels of IL-1β, purinergic receptor P2X7 (P2X7R), and phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK). ResultCompared with that in the normal group, the pain threshold of rats in the model group was significantly lower (P<0.01). The absolute value of neutrophils (NEUT#) and the percentage of neutrophils (NEUT) were significantly improved, and the percentage of lymphocytes (LYMPH) was significantly reduced (P<0.01). The serum levels of IL-1, IL-6, IL-8, and TNF-α were significantly increased (P<0.01). SP content in brain tissue was significantly increased, and β-EP content was significantly decreased (P<0.01). The relative protein expression of IL-1β, P2X7R, and p-p38 MAPK was significantly increased (P<0.05). HE staining and transmission electron microscopy results of medulla oblongata tissue revealed neuronal degeneration, mild proliferation of microglial cells, reduction in the number of myelinated nerves, and obvious demyelination. The trigeminal nerve fibers of rats were disarranged, and some nerve fibers showed vacuolization. Axons were swollen, and Schwann cells proliferated. Demyelination was observed. Compared with the model group, each administration group significantly increased the pain threshold of rats (P<0.05, P<0.01), reduced NEUT# and NEUT, and elevated LYMPH (P<0.05, P<0.01). The administration group significantly decreased the levels of IL-1, IL-6, IL-8, and TNF-α in serum and SP in brain tissue (P<0.01) and increased the level of β-EP (P<0.01). They significantly down-regulated the protein expression of IL-1β, P2X7R, and p-p38 MAPK(P<0.05, P<0.01) and significantly ameliorated the pathological changes in medulla oblongata tissue and trigeminal nerves of rats. ConclusionLutongning Granules had significant therapeutic effects on trigeminal neuralgia induced by injection of talc into the infraorbital foramen of model rats, and the mechanism may be related to amelioration of P2X7R-mediated neuroinflammation and inhibition of demyelination of myelinated nerves.
5.Prevention and treatment of monkeypox in pregnant women
Yeqin DENG ; Ying HONG ; Bin ZHANG ; Yang GAO ; Yanming WAN ; Ronghua XU ; Yaling ZONG
Journal of Public Health and Preventive Medicine 2024;35(5):130-133
Using China National Knowledge Infrastructure, Wanfang, and PubMed database, literature search was conducted with the keywords “pregnancy” and “monkeypox”, and 27 related research articles were selected for analysis. Through a comprehensive review of the related literature, we aim to improve our knowledge of this viral disease, better our prevention, treatment and responses to future monkeypox outbreaks in China, so as to better protect the safety of mothers and infants. Maternal monkeypox can be prevented and controlled, if active and effective measures are taken in time. Drawing on the experience and lessons from monkeypox outbreaks at home and abroad, it is suggested that hospitals and public health agencies at all levels should raise awareness, and establish an effective emergency preparedness system for the prevention and control of potential future outbreaks.
6.Malnutrition among primary and secondary school students from 2008 to 2021 in Zhejiang Province
YANG Ying, ZHENG Weijun, GU Fang, MENG Jia, LI Juanjuan, GAO Lei, SHEN Yu, ZHANG Ronghua
Chinese Journal of School Health 2024;45(9):1255-1259
Objective:
To describe the prevalence characteristics and trend of malnutrition among primary and secondary school students in Zhejiang Province from 2008 to 2021, so as to provide scientific references for targeted interventions on malnutrition among children and adolescents.
Methods:
Based on the National Student Common Diseases and Health Influencing Factors Surveillance Project, 81 228 primary and middle school students aged 9-17 from Zhejiang Province were recruited for a questionnaire in 2008, 2014 and 2021, with stratified cluster random sampling method. Malnutrition was determined by Screening for Malnutrition in School aged Children and Adolescents. The Kruskal-Wallis test was used for non normally distributed data, and the Chi square test was used for categorical data. A trend Chi square test analyzed detection rates across different years.
Results:
The prevalence rates of malnutrition, stunting, mild wasting, and moderate to severe wasting among primary and secondary school students in 2008, 2014 and 2021 were 12.0%, 6.6%, 6.0%; 1.2%, 0.5%, 0.3%; 6.0%, 3.9%, 3.6%; and 4.7%, 2.2%, 2.2%, with an overall declining trend ( χ 2 trend =532.73, 181.43, 161.24, 240.38, P <0.05). The prevalence rates of malnutrition and mild wasting in each year were higher in boys (13.7%, 7.6%; 7.5%, 5.0%; 7.1%, 4.5%) than in girls (10.3%, 4.4%; 5.5%, 2.7%; 4.8%, 2.5%) ( χ 2=54.45, 88.67; 47.04, 81.07; 85.28, 98.81; P <0.01). The difference in malnutrition prevalence between urban and rural areas gradually narrowed (12.5%, 11.6%; 6.9%, 6.3%; 6.0%, 6.0%), with no statistically significant difference in 2021 ( χ 2= 0.01 , P >0.05). Malnutrition among primary and secondary school students was primarily characterized by mild wasting (56.0%) in Zhejiang Province. Compared to 2008, the prevalence of malnutrition in 2014 and 2021 showed a steady upward trend with increasing age in Zhejiang Province( χ 2 trends =44.52, 11.78, P <0.01).
Conclusions
The prevalence of malnutrition among primary and secondary school students aged 9 to 17 years in Zhejiang Province decreases by year from 2008 to 2021. However, the prevalence increase steadily with age, and boys have a higher prevalence of malnutrition. Policies should be developed age , gender , and growth appropriate dietary patterns to reduce malnutrition among primary and secondary school students.
7.Effect and Mechanism of Shuangshenling Granules on Chronic Renal Failure Rat Model
Shuran LI ; Yanying LI ; Jingsheng ZHANG ; Ronghua ZHAO ; Shanshan GUO ; Qiyue SUN ; Zihan GENG ; Lei BAO ; Shuangrong GAO ; Xiaolan CUI ; Jing SUN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):46-55
ObjectiveTo investigate the pharmacological action and mechanism of Shuangshenling granules in treating chronic renal failure in rats,providing laboratory data to support clinical application of Shuangshenling granules. MethodSD rats (150-180 g),half males and half females in number,were used,with ten rats designated as the normal group,ten as the sham operation group,and the remaining rats undergoing chronic renal failure modeling induced by 5/6 nephrectomy. Two weeks after operation,serum creatinine (SCr) and blood urea nitrogen (BUN) levels were measured via orbital blood sampling to select successful model rats. Based on SCr values,the rats were evenly divided into the model group,Shenshuaining positive group (0.84 g·kg-1·d-1),and high,medium,and low dose groups of Shuangshenling granules (4.8,2.4,1.2 g·kg-1·d-1),with ten animals in each group. Each treatment group received drugs at 10 mL·kg-1 via intragastric administration once daily for six weeks. At 2,4,6 weeks after administration,SCr,BUN,24-hour urine volume,total urinary protein (UTP),urinary creatinine (UCr),creatinine clearance rate (CCr),serum albumin (SAlb),and total serum protein (STP) were measured. Following the experiment,kidney tissues were dissected for pathological examination. The expression levels of autophagy-related proteins,including PTEN-induced kinase 1 (PINK1),E3 ubiquitin-protein ligase parkin (Parkin),and microtubule-associated protein 1 light chain 3B (LC3B),were detected by immunofluorescence. ResultCompared with the normal group,the model group exhibited significantly increased levels of SCr,BUN,24-hour urine volume,UTP,and UCr (P<0.01),and decreased levels of SAlb and STP (P<0.01). CCr showed an initial increase followed by a decrease. Histopathological results revealed glomerular hyperplasia and atrophy,with varying degrees of mesangial cell reduction,blood stasis in the glomeruli,and significant widening of Bowman's capsule. Visceral parietal layer cells were displaced or absent,leading to incomplete and damaged glomeruli. A large number of protein casts were present in the proximal and distal convoluted tubules,with reduced and displaced cells,swelling in some tubules,and interstitial inflammatory exudation predominantly comprising lymphocytes and a small number of neutrophils. Compared with the model group,all dose groups of Shuangshenling granules significantly reduced levels of SCr,BUN,24-hour urine volume,UTP,and UCr (P<0.05,P<0.01) and increased SAlb and STP levels (P<0.01) at 2,4,and 6 weeks after administration. The three dose groups also improved CCr and alleviated renal pathological injury in varying degrees at 2-6 weeks after administration. Immunofluorescence results showed that the expression levels of PINK1,Parkin,and LC3B were significantly reduced in the model group compared with the normal group,whereas all dose groups of Shuangshenling granules significantly upregulated the expression levels of PINK1,Parkin,and LC3B compared with the model group. ConclusionShuangshenling granules significantly improved renal function and pathological injury in rats with chronic renal failure,likely through the upregulation of PINK1-mediated autophagy.
8.Influencing factors for abnormal spinal curvature among middle school students in Zhejiang Province
LI Juanjuan ; ZHANG Ronghua ; MENG Jia ; GAO Lei ; SHEN Yu ; GU Fang ; ZHU Bing
Journal of Preventive Medicine 2024;36(12):1036-1039,1044
Objective:
To investigate the status and influencing factors of abnormal spinal curvature among middle school students in Zhejiang Province, so as to provide insights into formulating intervention measures of abnormal spinal curvature among students.
Methods:
Middle school students were selected from 90 counties (cities or districts) for abnormal spinal curvature screening using the stratified random cluster sampling method based on the Student Common Diseases and Influencing Factors Program in 2022. Basic information, nutritional status, sedentary time and sleep time were collected through questionnaire surveys and physical examination. Influencing factors for abnormal spinal curvature among middle school students were identified using a multivariable logistic regression model.
Results:
Totally 93 988 students were recruited, and 92 559 students responded effectively, with a response rate of 98.48%. There were 48 578 males (52.48%) and 43 981 females (47.52%). A total of 1 690 students with abnormal spinal curvature were detected, accounting for 1.83%. Multivariable logistic regression analysis showed that the students who were female (OR=1.331, 95%CI: 1.204-1.471), lived in suburb areas (OR=1.254, 95%CI: 1.135-1.385), were at ages of 14 years and above (14 to 16 years, OR=1.252, 95%CI: 1.108-1.414; >16 years, OR=1.273, 95%CI: 1.090-1.486) and had sedentary time greater than 10 h/d (OR=1.162, 95%CI: 1.034-1.305) might have higher risks of abnormal spinal curvature, while the students who were overweight or obesity (OR=0.491, 95%CI: 0.426-0.565), regularly adjust the heights of the desks and chairs (OR=0.883, 95%CI: 0.797-0.979), and slept 8 hours or more per day (OR=0.850, 95%CI: 0.765-0.945) might have lower risks of abnormal spinal curvature.
Conclusions
The prevalence of abnormal spinal curvature among middle school students in Zhejiang Province is associated with gender, region, age, nutritional status, regularly adjusting the heights of desks and chairs, sedentary time and sleep time.
9.Construction and cross-protection of marker-free infC mutant of bovine Pasteu-rella multocida type A
Xuxia YANG ; Pei HU ; Jiaqi HUANG ; Lixu GAO ; Ronghua LYU ; Yangyang QIU ; Fang HE ; Yuanyi PENG ; Nengzhang LI
Chinese Journal of Veterinary Science 2024;44(9):1930-1939
infC gene encodes the translation initiation factor IF3 in bovine Pasteuella multocida,but it whether or not regulation to the virulence and cross-protection in P.multocida is still not well understood.In this study,the infC gene mutant(△infC)derived from bovine P.multocida type A strain CQ2 was constructed using by homologous recombination method.Compared with wild strain,the △infC showed significant increasing in biofilm formation,but the capsule produc-tion,virulence and bacterial loading in organs were significant decreased,and the IL-1β secretion of mouse peritoneal macrophage increased.Along with the infC gene deletion,the expression of genes related to capsule synthesis and LPS synthesis and transport were significantly down-regulated,while that of genes related to biofilm synthesis and outer membrane protein were significantly up-regulated.The inactivated vaccines of wild type and mutant were prepared and mice were immu-nized twice then challenged with wild type strains,respectively.The immuno-protection rate of△in fC inactivated vaccine against bovine P.multocida type A,B and F were 100.0%,83.3%and 0.0%,respectively,and the immuno-protection rate that against rabbit type P.multocida was 33.3%.The results indicated that infC gene could affect the virulence of P.multocida by regula-ting the production of capsule and the expressions of virulence related factors,and the deletion of infC gene conferred a certain cross-protection property of strains.This study provided a certain foundation for the development of P.multocida vaccine.
10.Therapeutic Effect of Gegentang Granules on Mouse Model with hCoV-229E Pneumonia and Hanshi Yidu Xifei Syndrome
Zihan GENG ; Yanyan BAO ; Lei BAO ; Ronghua ZHAO ; Jing SUN ; Yujing SHI ; Shanshan GUO ; Xiali TAO ; Yingjie GAO ; Xiaolan CUI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(19):34-41
ObjectiveTo determine the therapeutic effect of Gegentang granules on a disease-syndrome mouse model combining human coronavirus 229E (hCoV-229E) pneumonia with Hanshi Yidu Xifei syndrome in vivo. MethodMice were randomly divided into normal group, infection group, cold-dampness group, model group, chloroquine phosphate group (0.18 g·kg-1), interferon-α2b (IFN-α2b) group (1.83×106 U·kg-1), Gegentang granules high-dose and low-dose groups (6.6, 3.3 g·kg-1) with 10 mice in each group. Cold-dampness environment and hCoV-229E infection were used for modeling, and the general status and lung index of mice in each group were observed. The viral load in lung tissue was detected by real-time fluorescent quantitative polymerase chain reaction (Real-time PCR), the pathological changes in lung tissue were evaluated by hematoxylin-eosin (HE) staining, the levels of serum gastrointestinal hormones and inflammatory factors in lung tissue were detected by enzyme-linked immunosorbent assay (ELISA), and the percentage of peripheral blood lymphocytes was detected by flow cytometry. ResultComparing with model group, Gegentang granules could significantly alleviate the physical signs of Hanshi Yidu Xifei syndrome, including listlessness, weakness of limbs, sticky stool, etc. Comparing with model group, Gegentang granules high-dose group significantly reduced lung index, histopathological score of interstitial lung and bronchus, and the level of serum motilin (P<0.05, P<0.01), two doses of Gegentang granules could significantly increase the level of serum gastrin (P<0.05, P<0.01), the percentage of CD4+, CD8+ T lymphocytes in peripheral blood (P<0.05, P<0.01), and the level of tumor necrosis factor-α (TNF-α) in lung tissue was significantly decreased (P<0.01), and the level of interleukin-6 (IL-6) showed decreasing tendency. ConclusionGegentang granules has therapeutic effect on model mice. It can improve the appearance and behavior characterization, regulate the level of gastrointestinal hormones, decrease lung index and histopathological score, and possibly play an immunomodulatory role by inhibiting the expression of inflammatory cytokines in lung tissue and restoring the percentage of peripheral blood lymphocytes.


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