1.Role of miR-140-5p/BCL2L1 in apoptosis and autophagy of HFOB1.19 and effect of Bushen Jianpi Huoxue Decoction.
Tong-Ying CHEN ; Sai FU ; Xiao-Yun LI ; Shu-Hua LIU ; Yi-Fu YANG ; Dong-Sheng YANG ; Yun-Jie ZENG ; Yang-Bo LI ; Dan LUO ; Hong-Xing HUANG ; Lei WAN
China Journal of Chinese Materia Medica 2025;50(3):583-589
Osteoporosis(OP) is a senile bone disease characterized by an imbalance between bone remodeling and bone formation. Targeting pathogenesis of kidney deficiency, spleen deficiency, and blood stasis, Bushen Jianpi Huoxue Decoction has a significant effect on the treatment of OP by tonifying kidney, invigorating spleen, and activating blood circulation. MicroRNA(miRNA) and the anti-apoptotic protein B-cell lymphoma-2-like protein 1(BCL2L1) are closely related to bone cell metabolism. Therefore, in this study, the binding of miR-140-5p to BCL2L1 was detected by dual luciferase assay and polymerase chain reaction(PCR). After silencing or overexpressing miR-140-5p, the apoptosis, autophagy, and osteogenic function of human fetal osteoblast cell line 1.19(HFOB1.19) were observed by flow cytometry and Western blot. Bushen Jianpi Huoxue Decoction-containing serum was prepared by intragastric administration of Bushen Jianpi Huoxue Decoction in rats. Different concentrations of Bushen Jianpi Huoxue Decoction-containing serum were used to treat HFOB1.19 with or without miR-140-5p mimic. The expression of osteogenic proteins in each group was observed, and the role of miR-140-5p/BCL2L1 in apoptosis and autophagy of HFOB1.19 was studied, along with the effect of Bushen Jianpi Huoxue Decoction on these processes. As indicated by the dual luciferase assay, miR-140-5p bound to BCL2L1. Flow cytometry and Western blot showed that miR-140-5p promoted apoptosis and inhibited autophagy in HFOB1.19. After intervention with high, medium, and low doses of Bushen Jianpi Huoxue Decoction-medicated serum, compared with the miR-140-5p NC group, the expression of osteocalcin(OCN), osteopontin(OPN), Runt-related transcription factor 2(RUNX2), and transforming growth factor beta 1(TGF-β1) decreased in the miR-140-5p mimic group, while the expression of bone morphogenetic protein 2(BMP2) showed no significant difference under high-dose intervention. Therefore, miR-140-5p/BCL2L1 can promote apoptosis and inhibit autophagy in HFOB1.19. Bushen Jianpi Huoxue Decoction can affect the osteogenic effect of miR-140-5p through BMP2.
MicroRNAs/metabolism*
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Autophagy/drug effects*
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Apoptosis/drug effects*
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Humans
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Drugs, Chinese Herbal/administration & dosage*
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Animals
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Cell Line
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bcl-X Protein/metabolism*
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Osteoblasts/metabolism*
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Rats
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Osteoporosis/physiopathology*
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Male
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Rats, Sprague-Dawley
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Osteogenesis/drug effects*
2.Current situation of medicinal animal breeding and research progress in sustainable utilization of resources.
Cheng-Cai ZHANG ; Jia WANG ; Yu-Jie ZHOU ; Xiao-Yu DAI ; Xiu-Fu WAN ; Chuan-Zhi KANG ; De-Hua WU ; Jia-Hui SUN ; Sheng WANG ; Lan-Ping GUO
China Journal of Chinese Materia Medica 2025;50(16):4397-4406
Traditional Chinese medicine(TCM) is the pillar for the development of motherland medicine, and animal medicine has a long history of application in China, characterized by wide resources, strong activity, definite efficacy, and great benefits. It has significant potential and important status in the consumption market of raw materials of TCM. In the context of global climate change, farming system alterations, and low renewability, the depletion of wild medicinal animal resources has accelerated. Accordingly, the conservation and sustainable utilization of wild resources of animal medicinal materials has become a problem that garners increasing attention and urgently needs to be solved. This paper summarizes the current situation of domestic and foreign medicinal animal breeding and research progress in industrial application in recent years and points out the issues related to standardized breeding, germplasm selection and breeding, and quality evaluation standards for medicinal animals. Furthermore, this paper discusses standardized breeding, quality standards, resource protection and utilization, and the search for alternative resources for rare and endangered medicinal animals. It proposes that researchers should systematically carry out in-depth basic research on animal medicine, improve the breeding scale and level of medicinal animals, employ modern technology to enhance the quality standards of medicinal materials, and strengthen the research and development of alternative resources. This approach aims to effectively address the relationship between protection and utilization and make a significant contribution to the sustainable development of medicinal animal resources and the animal-based Chinese medicinal material industry.
Animals
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Breeding
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China
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Medicine, Chinese Traditional
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Conservation of Natural Resources
3.Associations of Ureaplasma urealyticum infection with male infertility and intrauterine insemination outcomes.
Yang-Yang WAN ; Xiao-Yun SHI ; Wen-Jing LIU ; Shun BAI ; Xin CHEN ; Si-Yao LI ; Xiao-Hua JIANG ; Li-Min WU ; Xian-Sheng ZHANG ; Juan HUA
Asian Journal of Andrology 2025;27(2):219-224
Ureaplasma urealyticum (UU) is one of the most commonly occurring pathogens associated with genital tract infections in infertile males, but the impact of seminal UU infection in semen on intrauterine insemination (IUI) outcomes is poorly understood. We collected data from 245 infertile couples who underwent IUI at The First Affiliated Hospital of USTC (Hefei, China) between January 2021 and January 2023. The subjects were classified into two groups according to their UU infection status: the UU-positive group and the UU-negative group. We compared semen parameters, pregnancy outcomes, and neonatal birth outcomes to investigate the impact of UU infection on IUI outcomes. There were no significantly statistical differences in various semen parameters, including semen volume, sperm concentration, total and progressive motility, sperm morphology, leukocyte count, the presence of anti-sperm antibody, and sperm DNA fragmentation index (DFI), between the UU-positive and UU-negative groups of male infertile patients (all P > 0.05). However, the high DNA stainability (HDS) status of sperm differed between the UU-positive and UU-negative groups, suggesting that seminal UU infection may affect sperm nuclear maturation ( P = 0.04). Additionally, there were no significant differences in pregnancy or neonatal birth outcomes between the two groups (all P > 0.05). These results suggest that IUI remains a viable and cost-effective option for infertile couples with UU infection who are facing infertility issues.
Humans
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Male
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Ureaplasma Infections/complications*
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Female
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Infertility, Male/therapy*
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Ureaplasma urealyticum/isolation & purification*
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Pregnancy
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Adult
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Pregnancy Outcome
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Semen Analysis
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Insemination, Artificial
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Semen/microbiology*
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China
4.Feasibility study on quantifying retinal vascular features for predicting preeclampsia based on artificial intelligence models
Tianfan ZHOU ; Feixue SHAO ; Sheng WAN ; Chenchen ZHOU ; Sijin ZHOU ; Xiaolin HUA
Journal of Shanghai Jiaotong University(Medical Science) 2024;44(5):552-559
Objective·To explore the predictive capability of retinal vascular features in preeclampsia(PE)based on artificial intelligence(AI)models.Methods·This retrospective study enrolled 789 pregnant women who registered from June 2020 to January 2021 at Shanghai First Maternity and Infant Hospital of Tongji University in the first 16 weeks of gestation.These women underwent regular prenatal examinations,had retinal fundus images captured,and delivered singleton live births at the hospital.According to whether they developed hypertensive disorders of pregnancy(HDP),they were divided into unaffected group(n=685)and HDP group(n=104).Within the HDP group,pregnancies were further categorized into gestational hypertension(GH)group(n=36)and PE group(n=68)based on the occurrence of PE.Based on the gestational age at onset,the PE group was further divided into early-onset PE group(gestational age<34 weeks)and late-onset PE group(gestational age≥34 weeks).Fundus images of the pregnant women were obtained,and an AI algorithm was utilized to diagnose retinal lesions and quantify retinal vascular features.Comparative analyses were conducted on fundus features and retinal vascular features.Univariate Logistic regression model was employed to analyze the influencing factors of PE occurrence,and multivariate Logistic regression model was further utilized to assess the correlation between retinal vascular features and the occurrence of PE.The predictive capability of retinal vascular features for PE(both early-and late-onset PE)was analyzed by using area under the curve(AUC)of receiver operator characteristic curve(ROC curve).Results·The comparative analysis of fundus features and retinal vascular features demonstrated statistically significant differences between the unaffected group and PE group in terms of central retinal artery equivalent(CRAE),central retinal vein equivalent(CRVE),arteriole-to-venular ratio(AVR),retinal artery tortuosity and retinal artery fractal dimension(all P<0.05).Univariate Logistic regression analysis indicated that second-trimester mean arterial pressure(MAP),second-trimester estimated fetal weight(EFW),CRAE,CRVE,AVR,retinal artery tortuosity and retinal artery fractal dimension were the influencing factors for PE occurrence(all P<0.05).Multivariate Logistic regression analysis revealed that second-trimester EFW,CRAE,CRVE,AVR,retinal artery tortuosity and retinal artery fractal dimension were the protective factors for the occurrence of PE,while second-trimester MAP was the risk factor for PE(all P<0.05).The analysis of ROC curves revealed that maternal risk factors along with second-trimester prenatal examination data(including MAP and EFW)and retinal vascular features model had good predictive ability for PE[AUC(95%CI)=0.784(0.725-0.843),and this model exhibited better predictive capability for early-onset PE,with an AUC(95%CI)of 0.840(0.756-0.924).Conclusion·The integration of quantified retinal vascular features based on AI models with maternal risk factors and second-trimester prenatal examination data(including MAP and EFW)enables a more effective prediction of PE occurrence,particularly early-onset PE.
5.Study on the Effect of Regulating DHPR/RyR Pathway by Pressing and Rubbing Method on the Improvement of Myofascial Pain Syndrome in Rats
Chao XIANG ; Sheng-Hua HE ; Xin ZHAO ; Qi WAN ; Chi MA ; Yan-Ping HU
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(5):1270-1276
Objective To explore the therapeutic effect and mechanism of pressing and rubbing method on myofascial pain syndrome(MPS)rats.Methods A total of 12 rats were randomly selected from 60 rats as the normal group,and the remaining rats were used to construct the MPS model by blunt strike combined with centrifugal exercise.Then 48 successfully modeled rats were randomly divided into model group,pressing and rubbing method group,pressing and rubbing method + Dantrolene[ryanodine receptor(RyR)inhibitor]group,pressing and rubbing method + normal saline group,with 12 rats in each group.The mechanical pain threshold was measured by von-Frey method.Detection of soft tissue tension,electromyography was performed;the ultrastructure of the pain point tissue was observed by transmission electron microscopy.The content of calcium ion(Ca2+)in the tissue of trigger point was detected by colorimetry.The protein expressions of dihydropyridine receptor(DHPR)α1,RyR and acetylcholinesterase(AChE)in the pain points of rats were detected by Western Blot.Results Compared with the normal group,the mechanical pain threshold,soft tissue tension in trigger point and the protein expressions of DHPRα1,RyR and AChE in the model group were decreased and the Ca2+ + content was increased(all P<0.05),and the peak potential with higher amplitude was observed in the electromyogram.The ultrastructure of the trigger point tissue was damaged.Compared with the model group,the mechanical pain threshold,soft tissue tension of trigger point and the protein expressions of DHPRα1,RyR and AChE in the trigger point tissue of the rats in the pressing and rubbing method group and the pressing and rubbing method + normal saline group were increased,and the Ca2+ content was decreased(all P<0.05),the electromyography was restored to be stable,the ultrastructural damage of the trigger point tissue was alleviated.Compared with the pressing and rubbing method group,the mechanical pain threshold,soft tissue tension of trigger point and protein the expressions of DHPRα1,RyR and AChE in the trigger point tissue of the rats in the pressing and rubbing method + Dantrolene group were decreased,and the Ca2+ content was increased(all P<0.05),the electromyogram showed electrical activity changes,the ultrastructure of the trigger point tissue was damaged.Conclusion The pressing and rubbing method may improve MPS in rats by activating the DHPR/RyR signaling pathway.
6.Clinical characteristics of 267 children with eosinophilic gastrointestinal disease:a multicenter study
Chun-Lei ZHAN ; Jie-Yu YOU ; Xiao-Qin LI ; Yong WANG ; Xian-Qin MEI ; Sheng-Hua WAN
Chinese Journal of Contemporary Pediatrics 2024;26(2):139-144
Objective To explore the clinical manifestations,endoscopic findings,histopathological changes,treatment,and prognosis of eosinophilic gastrointestinal disease(EGID)in children,with the aim of enhancing awareness among pediatricians about this condition.Methods Data of 267 children with EGID were prospectively collected from January 2019 to July 2022 at Jiangxi Children's Hospital,Hunan Children's Hospital,and Henan Children's Hospital.The age of onset,symptoms,physical signs,laboratory examination results,endoscopic findings,histopathological changes,and treatment outcomes were observed.Results Among the 267 children with EGID,the majority had mild(164 cases,61.4% )or moderate(96 cases,35.6% )clinical severity.The disease occurred at any age,with a higher prevalence observed in school-age children(178 cases).The main symptoms in infants were vomiting and hematemesis,while in toddlers,vomiting and bloody stools were prominent.Abdominal pain and vomiting were the primary symptoms in preschool and school-age children.Nearly half(49.4% )of the affected children showed elevated platelet counts on hematological examination,but there was no significant difference in platelet counts among children with mild,moderate,and severe EGID(P>0.05).Endoscopic findings in EGID children did not reveal significant specificity,and histopathological examination showed no specific structural damage.Among them,85.0% (227 cases)received acid suppression therapy,34.5% (92 cases)practiced dietary avoidance,20.9% (56 cases)received anti-allergic medication,and a small proportion(24 cases,9.0% )were treated with prednisone.Clinical symptoms were relieved in all patients after treatment,but three cases with peptic ulcers experienced recurrence after drug discontinuation.Conclusions Mild and moderate EGID are more common in children,with no specific endoscopic findings.Dietary avoidance,acid suppression therapy,and anti-allergic medication are the main treatment methods.The prognosis of EGID is generally favorable in children.[Chinese Journal of Contemporary Pediatrics,2024,26(2):139-144]
7.A multicenter study of neonatal stroke in Shenzhen,China
Li-Xiu SHI ; Jin-Xing FENG ; Yan-Fang WEI ; Xin-Ru LU ; Yu-Xi ZHANG ; Lin-Ying YANG ; Sheng-Nan HE ; Pei-Juan CHEN ; Jing HAN ; Cheng CHEN ; Hui-Ying TU ; Zhang-Bin YU ; Jin-Jie HUANG ; Shu-Juan ZENG ; Wan-Ling CHEN ; Ying LIU ; Yan-Ping GUO ; Jiao-Yu MAO ; Xiao-Dong LI ; Qian-Shen ZHANG ; Zhi-Li XIE ; Mei-Ying HUANG ; Kun-Shan YAN ; Er-Ya YING ; Jun CHEN ; Yan-Rong WANG ; Ya-Ping LIU ; Bo SONG ; Hua-Yan LIU ; Xiao-Dong XIAO ; Hong TANG ; Yu-Na WANG ; Yin-Sha CAI ; Qi LONG ; Han-Qiang XU ; Hui-Zhan WANG ; Qian SUN ; Fang HAN ; Rui-Biao ZHANG ; Chuan-Zhong YANG ; Lei DOU ; Hui-Ju SHI ; Rui WANG ; Ping JIANG ; Shenzhen Neonatal Data Network
Chinese Journal of Contemporary Pediatrics 2024;26(5):450-455
Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75%(27/36);boys accounted for 64%(23/36).Among the 36 neonates,31(86%)had disease onset within 3 days after birth,and 19(53%)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61%)had left cerebral infarction and 13(36%)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75%)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72%)and seizures in 10 neonates(34%).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33%,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44%(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.
8.Metformin and statins reduce hepatocellular carcinoma risk in chronic hepatitis C patients with failed antiviral therapy
Pei-Chien TSAI ; Chung-Feng HUANG ; Ming-Lun YEH ; Meng-Hsuan HSIEH ; Hsing-Tao KUO ; Chao-Hung HUNG ; Kuo-Chih TSENG ; Hsueh-Chou LAI ; Cheng-Yuan PENG ; Jing-Houng WANG ; Jyh-Jou CHEN ; Pei-Lun LEE ; Rong-Nan CHIEN ; Chi-Chieh YANG ; Gin-Ho LO ; Jia-Horng KAO ; Chun-Jen LIU ; Chen-Hua LIU ; Sheng-Lei YAN ; Chun-Yen LIN ; Wei-Wen SU ; Cheng-Hsin CHU ; Chih-Jen CHEN ; Shui-Yi TUNG ; Chi‐Ming TAI ; Chih-Wen LIN ; Ching-Chu LO ; Pin-Nan CHENG ; Yen-Cheng CHIU ; Chia-Chi WANG ; Jin-Shiung CHENG ; Wei-Lun TSAI ; Han-Chieh LIN ; Yi-Hsiang HUANG ; Chi-Yi CHEN ; Jee-Fu HUANG ; Chia-Yen DAI ; Wan-Long CHUNG ; Ming-Jong BAIR ; Ming-Lung YU ;
Clinical and Molecular Hepatology 2024;30(3):468-486
Background/Aims:
Chronic hepatitis C (CHC) patients who failed antiviral therapy are at increased risk for hepatocellular carcinoma (HCC). This study assessed the potential role of metformin and statins, medications for diabetes mellitus (DM) and hyperlipidemia (HLP), in reducing HCC risk among these patients.
Methods:
We included CHC patients from the T-COACH study who failed antiviral therapy. We tracked the onset of HCC 1.5 years post-therapy by linking to Taiwan’s cancer registry data from 2003 to 2019. We accounted for death and liver transplantation as competing risks and employed Gray’s cumulative incidence and Cox subdistribution hazards models to analyze HCC development.
Results:
Out of 2,779 patients, 480 (17.3%) developed HCC post-therapy. DM patients not using metformin had a 51% increased risk of HCC compared to non-DM patients, while HLP patients on statins had a 50% reduced risk compared to those without HLP. The 5-year HCC incidence was significantly higher for metformin non-users (16.5%) versus non-DM patients (11.3%; adjusted sub-distribution hazard ratio [aSHR]=1.51; P=0.007) and metformin users (3.1%; aSHR=1.59; P=0.022). Statin use in HLP patients correlated with a lower HCC risk (3.8%) compared to non-HLP patients (12.5%; aSHR=0.50; P<0.001). Notably, the increased HCC risk associated with non-use of metformin was primarily seen in non-cirrhotic patients, whereas statins decreased HCC risk in both cirrhotic and non-cirrhotic patients.
Conclusions
Metformin and statins may have a chemopreventive effect against HCC in CHC patients who failed antiviral therapy. These results support the need for personalized preventive strategies in managing HCC risk.
9.Bioinformatics and animal experiments reveal mechanism of Linggui Zhugan Decoction in ameliorating chronic heart failure after myocardial infarction via HIF-1α/HO-1 signaling pathway.
Han REN ; Shu-Shu WANG ; Wan-Zhu ZHAO ; Shao-Hua XU ; Ke-Dong WEI ; Wan-Wan WU ; Sheng-Yi HUANG ; Rui CAI ; Yuan-Hong ZHANG ; Jin-Ling HUANG
China Journal of Chinese Materia Medica 2024;49(23):6407-6416
This study aims to investigate the effect of Linggui Zhugan Decoction(LGZGD) on autophagy in the mouse model of chronic heart failure(CHF) induced by myocardial infarction(MI), as well as the regulatory effect of LGZGD on the hypoxia-inducible factor-1α(HIF-1α)/heme oxygenase-1(HO-1) signaling pathway, based on bioinformatics and animal experiments. The active ingredients and corresponding targets of LGZGD were retrieved from the Traditional Chinese Medicine Systems Pharmacology and Analysis Database, and GEO, GeneCards, and DisGeNET were searched for the disease targets. Cytoscape was used to establish a "drug-component-target" network. The protein-protein interaction(PPI) network analysis was performed on STRING. R language was used for Gene Ontology(GO) and Kyoto Encycloperfia of Genes and Genomes(KEGG) enrichment analyses. Molecular docking was adopted to validate the core targets. The mouse model of MI-induced CHF was established by surgical ligation of the left anterior descending coronary artery. The modeled mice were assigned into the sham, model, low-, medium-, and high-dose(2.34, 4.68, and 9.36 g·kg~(-1), respectively) LGZGD, and captopril(3.25 mg·kg~(-1)) groups. After continuous administration for 6 weeks, a Doppler ultrasound imaging system was used to examine the heart function indicators: left ventricular ejection fraction(LVEF), left ventricular fractional shortening(LVFS), left ventricular end-systolic dimension(LVIDs), and left ventricular end-diastolic dimension(LVIDd). The myocardial tissue was stained with hematoxylin-eosin for the observation of morphological changes. The mRNA levels of microtubule-associated protein 1 light chain 3 beta(LC3B), Beclin1, p62, HIF-1α, and HO-1 in the myocardial tissue were determined by RT-qPCR. The protein levels of LC3B, beclin1, p62, autophagy-related protein 5(ATG5), HIF-1α, and HO-1 were determined by Western blot. The results showed that 103 active components of LGZGD, corresponding to 224 targets, were obtained. A total of 3 485 and 6 165 targets related to MI and CHF, respectively, were retrieved. The GSE16499 dataset obtained 3 263 differentially expressed genes. There were 31 common targets. The top 3 core active components were quercetin, naringenin, and 1-methoxyphaseollidin. The topology analysis results showed that the core targets were MAPK3, HMOX1(HO-1), MYC, ADRB2, PPARD, and HIF1A(HIF-1α). The molecular docking results showed strong binding between the core targets and the main active components of LGZGD. LGZGD significantly improved the heart function and alleviated the pathological changes in the myocardial tissue of mice. Western blot and RT-qPCR results showed that the HIF-1α/HO-1 signaling pathway and autophagy were activated in the model group. LGZGD up-regulated the levels of LC3B, Beclin1, ATG5, HIF-1α, and HO-1 while down-regulating the mRNA and protein levels of p62. In summary, LGZGD can enhance autophagy and improve the heart function in the mouse model of CHF after MI by upregulating the HIF-1α/HO-1 signaling pathway.
Animals
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Drugs, Chinese Herbal/chemistry*
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Myocardial Infarction/drug therapy*
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Heart Failure/physiopathology*
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Mice
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Hypoxia-Inducible Factor 1, alpha Subunit/genetics*
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Signal Transduction/drug effects*
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Male
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Computational Biology
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Heme Oxygenase-1/genetics*
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Molecular Docking Simulation
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Protein Interaction Maps/drug effects*
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Mice, Inbred C57BL
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Humans
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Chronic Disease
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Disease Models, Animal
10.Primary study of heat stress inducing early expression and secretion of tissue factor in vascular endothelial cells
Jing QIAN ; Fan-Fan WANG ; Lu-Lu WAN ; Jia-Le YANG ; Xue-Zhi SHI ; Hua-Sheng TONG ; Wei-Liang ZHOU
Medical Journal of Chinese People's Liberation Army 2023;48(12):1412-1419
Objective To explore the pattern of early expression and secretion of tissue factor(TF)in vascular endothelial cells induced by heat stress.Methods Thirty SPF-rated C57BL/6 male mice were randomly divided into five groups:the control group and groups of indicated recovery time,including 0,3,6,and 9 h in room temperature after heat stress(n=6).Mice in the heat stress groups were exposed to an animal incubator to reach 42.5℃for core body temperature for heat stroke.We analyzed the histopathological changes in the liver,lung,and kidney tissues with HE staining.We measured the TF mRNA in mice tissues by RT-qPCR and the plasma concentration of TF in mice with a commercial ELISA kit.Human umbilical vein endothelial cells(HUVECs)were placed in a culture incubator to build an in vitro heat stress model.HUVECs were divided into five groups,including a control group and groups of indicated recovery time,including 0,3,6,and 9 h after heat stress.We quantified the expression of TF mRNA and protein in HUVEC cells by RT-qPCR,Western blotting,and immunofluorescence and measured the secreted TF with a commercial ELISA kit.Results No significant pathological injury was observed in the tissues of the control group.Mice treated with heat stress had various degrees of structural injuries and hemorrhagic and inflammatory changes in multiple tissues.Compared to control group,the expression of TF mRNA significantly increased in the kidney of heat stress-treated mice with 0 and 3 h recovery time(1.719±0.018,1.241±0.178 vs.1.000±0.063),the lung with 3 h recovery time(2.444±0.511 vs.1.000±0.106)and the liver with 6 h recovery time(7.312±0.618 vs.1.000±0.147)(P<0.05).The concentration of TF in plasma also sustainedly elevated in mice with 0,3,6,and 9 h recovery time after heat stress as compared to control group[(132.426±17.920)pg/ml,(119.400±10.267)pg/ml,(107.374±13.495)pg/ml,(163.767±22.810)pg/ml vs.(75.479±13.831)pg/ml,respectively,P<0.01].The expression levels of TF mRNA were higher in heat stress HUVECs with 6 h and 9 h recovery time than the control cells(1.905±0.354,2.564±0.297 vs.1.000±0.097,P<0.01).Secreted TF in the supernatant from HUVECs treated with heat stress and different recovery time also increased significantly[(36.309±4.101)pg/ml,(38.425±5.484)pg/ml,(41.655±4.380)pg/ml,(43.586±4.718)pg/ml vs.(14.996±0.254)pg/ml,P<0.01].Conclusion Heat stress increased early expression and secretion of TF in vascular endothelial cells.Vascular endothelial cells may be a main source of circulating TF in heat stroke.

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