1.Mechanism of salvianolic acid B protecting H9C2 from OGD/R injury based on mitochondrial fission and fusion
Zi-xin LIU ; Gao-jie XIN ; Yue YOU ; Yuan-yuan CHEN ; Jia-ming GAO ; Ling-mei LI ; Hong-xu MENG ; Xiao HAN ; Lei LI ; Ye-hao ZHANG ; Jian-hua FU ; Jian-xun LIU
Acta Pharmaceutica Sinica 2024;59(2):374-381
This study aims to investigate the effect of salvianolic acid B (Sal B), the active ingredient of Salvia miltiorrhiza, on H9C2 cardiomyocytes injured by oxygen and glucose deprivation/reperfusion (OGD/R) through regulating mitochondrial fission and fusion. The process of myocardial ischemia-reperfusion injury was simulated by establishing OGD/R model. The cell proliferation and cytotoxicity detection kit (cell counting kit-8, CCK-8) was used to detect cell viability; the kit method was used to detect intracellular reactive oxygen species (ROS), total glutathione (t-GSH), nitric oxide (NO) content, protein expression levels of mitochondrial fission and fusion, apoptosis-related detection by Western blot. Mitochondrial permeability transition pore (MPTP) detection kit and Hoechst 33342 fluorescence was used to observe the opening level of MPTP, and molecular docking technology was used to determine the molecular target of Sal B. The results showed that relative to control group, OGD/R injury reduced cell viability, increased the content of ROS, decreased the content of t-GSH and NO. Furthermore, OGD/R injury increased the protein expression levels of dynamin-related protein 1 (Drp1), mitofusions 2 (Mfn2), Bcl-2 associated X protein (Bax) and cysteinyl aspartate specific proteinase 3 (caspase 3), and decreased the protein expression levels of Mfn1, increased MPTP opening level. Compared with the OGD/R group, it was observed that Sal B had a protective effect at concentrations ranging from 6.25 to 100 μmol·L-1. Sal B decreased the content of ROS, increased the content of t-GSH and NO, and Western blot showed that Sal B decreased the protein expression levels of Drp1, Mfn2, Bax and caspase 3, increased the protein expression level of Mfn1, and decreased the opening level of MPTP. In summary, Sal B may inhibit the opening of MPTP, reduce cell apoptosis and reduce OGD/R damage in H9C2 cells by regulating the balance of oxidation and anti-oxidation, mitochondrial fission and fusion, thereby providing a scientific basis for the use of Sal B in the treatment of myocardial ischemia reperfusion injury.
2.The method of determination for 2, 3-Butanedione in the air of workplace by high performance liquid chromatography with derivatization
Haipeng YE ; Hong FU ; Ji SHAO ; Xiaoyue SHAN ; Ling ZHANG ; Lei ZHANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(2):129-132
Objective:To establish a method for the determination of 2, 3-Butanedione (BUT) in the air of workplace, which including the process of collection by absorption in phosphoric acid aqueous solution and the process of analysis and detection by high performance liquid chromatography with derivatization.Methods:In October 2022, a porous glass plate absorption tube containing 10 ml of 0.01% phosphoric acid solution was used to collect BUT in the air of the workplace at a flow rate of 0.2 L/min. The absorption solution was derived by 2, 4-dinitrophenylhydrazine for 75 min and separated on a SB-C18 column (250 mm×4.6 mm, 5 μm) . At the column temperature of 30 ℃, the mixture of acetonitrile-water ( V∶ V, 1∶1) was eluted at the flow rate of 1.0 ml/min. It was detected by UV detector (λ=365 nm) , qualitatived by retention time and quantitatived by external standard. Results:It showed that BUT in phosphoric acid aqueous solution could be stored for at least 7 d at 4 ℃. There was a linear relationship within the determination range of 0.05-6.00 μg/ml, the linear regression equation was y=89.610 x+0.133, r=0.9999. The sampling absorption efficiencies were 98.33%-100.00%, the detection limit of the method was 0.005 μg/ml, the minimum detection concentration was 0.016 mg/m 3 (based on V0=3.0 L) . The recovery rates were 95.96%-102.44%, the intra batch precision were 4.36%-7.78%, and the inter batch precision were 4.96%-6.06%. Conclusion:The method has the advantages of simple operation, high sensitivity and good accuracy. It can prevent the loss and degradation of BUT. It can be used for the determination of BUT in the air of workplace.
3.Research progresses of endogenous vascular calcification inhibitor BMP-7
Xin ZHOU ; Lu XING ; Peng-Quan LI ; Dong ZHAO ; Hai-Qing CHU ; Chun-Xia HE ; Wei QIN ; Hui-Jin LI ; Jia FU ; Ye ZHANG ; Li XIAO ; Hui-Ling CAO
Chinese Pharmacological Bulletin 2024;40(7):1226-1230
Vascular calcification is a highly regulated process of ectopic calcification in cardiovascular system while no effective intervention can be clinically performed up to date.As vascular calcification undergoes a common regulatory mechanism within bone formation,bone morphogenetic protein 7(BMP-7)main-tains contractile phenotype of vascular smooth muscle cells and further inhibits vascular calcification via promoting the process of osteoblast differentiation,reducing ectopic calcification pressure by increasing bone formation and reducing bone resorption.This work systematically reviews the role of BMP-7 in vascular calcifi-cation and the possible mechanism,and their current clinical application as well.The current proceedings may help develope early diagnostic strategy and therapeutic treatment with BMP-7 as a new molecular marker and potential drug target.The expec-tation could achieve early prevention and intervention of vascular calcification and improve poor prognosis on patients.
4.The method of determination for 2, 3-Butanedione in the air of workplace by high performance liquid chromatography with derivatization
Haipeng YE ; Hong FU ; Ji SHAO ; Xiaoyue SHAN ; Ling ZHANG ; Lei ZHANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(2):129-132
Objective:To establish a method for the determination of 2, 3-Butanedione (BUT) in the air of workplace, which including the process of collection by absorption in phosphoric acid aqueous solution and the process of analysis and detection by high performance liquid chromatography with derivatization.Methods:In October 2022, a porous glass plate absorption tube containing 10 ml of 0.01% phosphoric acid solution was used to collect BUT in the air of the workplace at a flow rate of 0.2 L/min. The absorption solution was derived by 2, 4-dinitrophenylhydrazine for 75 min and separated on a SB-C18 column (250 mm×4.6 mm, 5 μm) . At the column temperature of 30 ℃, the mixture of acetonitrile-water ( V∶ V, 1∶1) was eluted at the flow rate of 1.0 ml/min. It was detected by UV detector (λ=365 nm) , qualitatived by retention time and quantitatived by external standard. Results:It showed that BUT in phosphoric acid aqueous solution could be stored for at least 7 d at 4 ℃. There was a linear relationship within the determination range of 0.05-6.00 μg/ml, the linear regression equation was y=89.610 x+0.133, r=0.9999. The sampling absorption efficiencies were 98.33%-100.00%, the detection limit of the method was 0.005 μg/ml, the minimum detection concentration was 0.016 mg/m 3 (based on V0=3.0 L) . The recovery rates were 95.96%-102.44%, the intra batch precision were 4.36%-7.78%, and the inter batch precision were 4.96%-6.06%. Conclusion:The method has the advantages of simple operation, high sensitivity and good accuracy. It can prevent the loss and degradation of BUT. It can be used for the determination of BUT in the air of workplace.
5.Mechanism of Buyang Huanwu Decoction in protecting ischemic myocardium by regulating platelet autophagy in rats with acute myocardial infarction.
Jia-Ming GAO ; Hao GUO ; Ye-Hao ZHANG ; Ling-Mei LI ; Gao-Jie XIN ; Zi-Xin LIU ; Yue YOU ; Yuan-Yuan CHEN ; Jian-Xun LIU ; Jian-Hua FU
China Journal of Chinese Materia Medica 2023;48(15):4156-4163
This study explored the effects of Buyang Huanwu Decoction(BYHWD) on platelet activation and differential gene expression after acute myocardial infarction(AMI). SD rats were randomly divided into a sham-operated group, a model group, a positive drug(aspirin) group, and a BYHWD group. Pre-treatment was conducted for 14 days with a daily oral dose of 1.6 g·kg~(-1) BYHWD and 0.1 g·kg~(-1) aspirin. The AMI model was established using the high ligation of the left anterior descending coronary artery method. The detection indicators included myocardial infarct size, heart function, myocardial tissue pathology, peripheral blood flow perfusion, platelet aggregation rate, platelet membrane glycoprotein CD62p expression, platelet transcriptomics, and differential gene expression. The results showed that compared with the sham-operated group, the model group showed reduced ejection fraction and cardiac output, decreased peripheral blood flow, and increased platelet aggregation rate and CD62p expression, and activated platelets. At the same time, TXB_2 content increased and 6-keto-PGF1α content decreased in serum. Compared with the model group, BYHWD increased ejection fraction and cardiac output, improved blood circulation in the foot and tail regions and cardiomyocytes arrangement, reduced myocardial infarct size and inflammatory infiltration, down-regulated platelet aggregation rate and CD62p expression, reduced serum TXB_2 content, and increased 6-keto-PGF1α content. Platelet transcriptome sequencing results revealed that BYHWD regulated mTOR-autophagy pathway-related genes in platelets. The differential gene expression levels were detected using real-time quantitative PCR. BYHWD up-regulated mTOR, down-regulated autophagy-related FUNDC1 and PINK genes, and up-regulated p62 gene expression. The results demonstrated that BYHWD could regulate platelet activation, improve blood circulation, and protect ischemic myocardium in AMI rats, and its mechanism is related to the regulation of the mTOR-autophagy pathway in platelets.
Rats
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Animals
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Rats, Sprague-Dawley
;
Drugs, Chinese Herbal/therapeutic use*
;
Myocardial Infarction/genetics*
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Myocardium/metabolism*
;
Aspirin/therapeutic use*
;
TOR Serine-Threonine Kinases/metabolism*
;
Membrane Proteins/metabolism*
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Mitochondrial Proteins
6.Determination of diacetyl in workplace air by high performance liquid chromatography using 4-nitro-o-phenylenediamine as precolumn derivatization
Haipeng YE ; Ji SHAO ; Siwei TAN ; Xiaoyue SHAN ; Ling ZHANG ; Hong FU ; Lei ZHANG
Journal of Environmental and Occupational Medicine 2023;40(9):1079-1084
Background Diacetyl (DC) is widely used in the food flavoring industry and excessive occupational exposure to DC can cause serious respiratory diseases. However, there is no corresponding national standard method for the determination of DC in the air of workplace. Objective To establish a method for the determination of DC in workplace air by high performance liquid chromatography using 4-nitro-o-phenylenediamine (NPDA) as precolumn derivatization. Methods DC in the air of workplace was collected by solution absorption method. This experiment used NPDA as the derivatization reagent. By adjusting acidity of solution and optimizing concentration ration of DC/NPDA, derivatization temperature, and time, a method for the determination of DC in workplace air was proposed, and its performance indexes such as linearity, detection limit, and lower limit of quantification were obtained. Sampling efficiency was evaluated by relative comparison method, and sample stability was evaluated by sample preservation test. Accuracy and precision of the method were evaluated by standard addition recovery test with blank samples, and an interference test was carried out by adding standard samples. The established method was applied to actual samples to evaluate its adaptability. Results A combination of 60 °C for 2 h was selected for derivatization because a higher derivatization reaction temperature and a longer reaction time associated with a higher derivatization efficiency. The solution was separated by SB-C18 column (250 mm×4.6 mm, 5 μm) at 30 ℃, using a mixture of methanol and water (v/v, 65%/35%) as mobile phase with an elution flow rate of 1.0 mL·min−1, and was detected with a variable wavelength detector (λmax=257 nm) by qualitative analysis based on retention time and quantitative analysis based on external standard method. In terms of the proposed method, the linear range of detection was from 5 μg·L−1 to 2000 μg·L−1, with a correlation coefficient of 0.9999, and a detection limit of 1.3 μg·L−1, the quantitative detection of the lower limit was 4.3 μg·L−1, with a sampling volume V0 of 3.0 L, the minimum detection concentration was 4.3 μg·m−3, and the minimum quantitative concentration was 14.3 μg·m−3. The recovery rate was 99.1%-100.8%, the intra-batch precision was 0.5%-3.0%, and the inter-batch precision was 1.2%-2.0%. The average sampling efficiency of this method was 94.5%, and the sample could be stored at 4 °C for at least 14 d. The coexisting components in the air of the workplace did not interfere with the determination of DC. The DC content in the air of a flavor workplace was 5.86-8.85 mg·m−3. Conclusion A determination method for DC in workplace air by high performance liquid chromatography using NPDA as precolumn derivatization after being collected by 1.0% phosphoric acid absorbent is proposed and has the advantages of simple operation, high sensitivity, and good accuracy. With no DC loss and degradation, the method may satisfy the request for DC determination in the air of workplace.
7.A Case Series of Olfactory Dysfunction in Imported COVID-19 Patients: A 12-Month Follow-Up Study.
Ni WANG ; Ming Bo YANG ; Pu Ye YANG ; Ren Bo CHEN ; Fei HUANG ; Nan Nan SHI ; Yan MA ; Yan ZHANG ; You XU ; Si Hong LIU ; Heng Yi LU ; Qing Qing FU ; Yi Pin FAN ; Hong Min KAN ; Xiao Hong WANG ; Ya Ling GUO
Biomedical and Environmental Sciences 2022;35(5):402-411
Objective:
The scientific community knows little about the long-term influence of coronavirus disease 2019 (COVID-19) on olfactory dysfunction (OD). With the COVID-19 pandemic ongoing worldwide, the risk of imported cases remains high. In China, it is necessary to understand OD in imported cases.
Methods:
A prospective follow-up design was adopted. A total of 11 self-reported patients with COVID-19 and OD from Xi'an No. 8 Hospital were followed between August 19, 2021, and December 12, 2021. Demographics, clinical characteristics, laboratory and radiological findings, and treatment outcomes were analyzed at admission. We surveyed the patients via telephone for recurrence and sequelae at the 1-, 6-, and 12-month follow-up.
Results:
Eleven patients with OD were enrolled; of these, 54.5% (6/11) had hyposmia and 45.5% (5/11) had anosmia. 63.6% (7/11) reported OD before or on the day of admission as their initial symptom; of these, 42.9% (3/7) described OD as the only symptom. All patients in the study received combined treatment with traditional Chinese medicine and Western medicine, and 72.7% (8/11) had partially or fully recovered at discharge. In terms of OD recovery at the 12-month follow-up, 45.5% (5/11) reported at least one sequela, 81.8% (9/11) had recovered completely, 18.2% (2/11) had recovered partially, and there were no recurrent cases.
Conclusions
Our data revealed that OD frequently presented as the initial or even the only symptom among imported cases. Most OD improvements occurred in the first 2 weeks after onset, and patients with COVID-19 and OD had favorable treatment outcomes during long-term follow-up. A better understanding of the pathogenesis and appropriate treatment of OD is needed to guide clinicians in the care of these patients.
COVID-19/complications*
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Follow-Up Studies
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Humans
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Olfaction Disorders/etiology*
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Pandemics
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Prospective Studies
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SARS-CoV-2
8.Effect of Ethyl Acetate Extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis on Intestinal Flora in ApoE-/- Mice with Atherosclerosis
Ai-ling JIA ; Yu-hang ZHANG ; Yuan-yuan DIAO ; Yu-wen SHI ; Meng-cheng JIANG ; Jun-ge LI ; Ye QIU ; Zhi-dong QIU ; Chao-mei FU
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(5):108-115
ObjectiveTo observe the effect of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis on high-fat diet-induced apolipoprotein E gene knockout (ApoE-/-) mice, and explore its mechanism of treating atherosclerosis by regulating intestinal flora. MethodThirty-two 8-week-old male ApoE-/- mice were randomly divided into model group, rosuvastatin group (10 mg·kg-1), high-, low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis (75, 25 mg·kg-1), with 8 mice in each group. Eight C57BL/6 mice were used as blank group. After 8 weeks of continuous administration, blood was taken to determine the blood lipid level. Enzyme-linked immunosorbent assay (ELISA) was used to detect the contents of related indexes in serum of mice. Hematoxylin-eosin (HE) staining was used to observe the formation of aortic plaque in mice. Cecal contents were collected and 16S rRNA amplicon sequencing was used to detect intestinal flora. ResultCompared with the blank group, the plaque area of the model group was significantly increased with inflammatory infiltration, the contents of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), inflammatory factors and inducible nitric oxide synthase (iNOS) were increased, while the content of high-density lipoprotein cholesterol (HDL-C) was decreased. Compared with the model group, rosuvastatin group and high- and low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could improve the deposition of aortic plaque, reduce the contents of TG, TC, LDL-C, inflammatory factors and iNOS, and increase the content of HDL-C. Compared with the blank group, the relative abundances of Firmicutes and Proteobacteria in the model group increased, while the relative abundance of Bacteroidetes decreased. Alpha and Beta diversity analysis showed that samples of each group could be significantly isolated, and the total number and abundance of intestinal flora species in the model group were low. Compared with the model group, ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could increase the relative abundance of beneficial bacteria and decrease the relative abundance of pathogenic bacteria. ConclusionEthyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis was mainly composed of flavonoids, which can treat atherosclerosis by regulating the intestinal flora and improve the pathological changes in the aorta of ApoE-/- mice induced by high-fat diet. The mechanism may be related to its ability to reduce the level of inflammatory factors, improve antioxidant capacity and repair the disorder of intestinal flora structure.
9.Evaluation of Dominant Diseases of Traditional Chinese Medicine: A Case Study of Ischemic Stroke
Wen-ran QIU ; Zhen-min XU ; Wei SHEN ; Guo-jing FU ; Liu-ding WANG ; Shao-jiao LIU ; Hong-xi LIU ; Yue LIU ; Ye-fei WANG ; Ling-ling DAI ; Chun-yan GUO ; Wei-wei JIAO ; Xing LIAO ; Yu LIU ; Xiao LIANG ; Yun-ling ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(12):225-232
The present study explored the effective approaches to realize the leading role of traditional Chinese medicine (TCM) in preventing diseases, the synergistic role in treating serious diseases, the core role in the rehabilitation of diseases and summarized the experience to provide feasible plans for the evaluation of other dominant diseases of TCM. To evaluate the effectiveness, safety, and economy of TCM in the treatment of ischemic stroke, encephalopathy project team of the China Center for Evidence-based Traditional Chinese Medicine(CCEBTCM) established an evaluation group to determine the work plan and complete the evaluation work. The concepts of the evaluation involved high-quality evidence, expert opinion survey, expert interview, and drug catalog. Under the guidance of clinical experts and methodologists, the evaluation work was completed in accordance with four steps, i.e., plan making, data collection and data extraction, evidence synthesis and evaluation, and report writing with the rapid review method. Through the review of TCM and western medicine experts, the advantage of TCM in the treatment of ischemic stroke was positioned in the convalescence period with the predominant effects of improving the neurological function defect and improving the daily living ability. In the convalescence period of stroke, TCM treatment could improve post-stroke motor dysfunction, post-stroke cognitive impairment, consciousness disorder, swallowing disorder, aphasia, constipation, urinary function, diplopia, etc., and the advantages of acupuncture, Chinese medicine, and traditional exercise were more prominent. In terms of safety, TCM treatment of ischemic stroke showed lower incidence of adverse reactions, fewer adverse events, and a milder degree of related symptoms. In terms of economic performance, the combined treatment of TCM and western medicine played a synergistic role and made the treatment cost more reasonable. Compared with conventional intervention, the integrated TCM and western medicine rehabilitation program showed more economic and social benefits.
10.Construction and effect of biobank in biological research in nursing field
Liang FU ; Xiaoyu LING ; Lin CHEN ; Lifen WU ; Lushan ZHENG ; Rufang LI ; Wenxia XU ; Yan HU ; Xianghong YE
Chinese Journal of Modern Nursing 2022;28(13):1727-1732
Objective:To explore the construction and effect of biobank in biological research in nursing field.Methods:Through the establishment of a biological research plan for nursing in Affiliated Jinhua Hospital, Zhejiang University School of Medicine, the biobank in biological research for nursing was constructed from aspects of biological samples, instruments and equipment, operating software, biobanking team formation, personnel training and standardized process formulation.Results:In August 2020, the Nursing Department and Central Laboratory of Affiliated Jinhua Hospital, Zhejiang University School of Medicine established a biobank for biological research in the nursing field, formed a biobank team, conducted personnel training and formulated standardized procedures. From December 1, 2020 to June 30, 2021, biological samples were collected from 264 gastric cancer patients and 56 healthy controls, mainly including whole blood, serum and urine.Conclusions:The construction of biobank for biological research in nursing field can collect abundant biological samples, which lays a foundation for subsequent independent biological research in nursing field.

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