1.Effect of Xidi Liangxue recipe on the proliferation and apoptosis of HaCaT cells through the lncRNA NEAT1/miR-485-5p/STAT3 regulatory network
Zhiming TANG ; Mengqing JING ; Lu LU ; Xiao SHAN ; Cuixia ZHANG ; Xiaoyu ZHANG ; Sa MENG
Chinese Journal of Dermatology 2023;56(7):642-650
Objective:To investigate the effect of Xidi Liangxue recipe on the proliferation and apoptosis of HaCaT cells through the long non-coding RNA (lncRNA) nuclear-enriched abundant transcript 1 (NEAT1) /microRNA (miR) -485-5p/signal transducer and activator of transcription 3 (STAT3) regulatory network. Methods:HaCaT cells were induced by interleukin-17 (IL-17), and the mRNA and protein expression of lncRNA NEAT1, miR-485-5p and STAT3 was detected in IL-17-induced HaCaT cells and normal human epidermal keratinocytes (NHEK) by quantitative PCR (qPCR) and Western blot analysis, respectively. The location of lncRNA NEAT1 and miR-485-5p in IL-17-induced HaCaT cells was observed by fluorescence in situ hybridization (FISH), and the targeted regulatory relationship among lncRNA NEAT1, miR-485-5p and STAT3 was verified by double-luciferase reporter gene assay. Chinese herbs were decocted according to the Xidi Liangxue recipe, SD rats were divided into two groups to be gavaged with the above decoctions (medicated group) or physiological saline (control group) for 5 days, and then serum samples were collected from the above two groups of rats separately. The IL-17-induced HaCaT cells were divided into 4 groups: control group treated with the control sera, lncRNA-NEAT1 overexpression group transfected with lncRNA-NEAT1 overexpression vectors and treated with the control sera, Xidi Liangxue recipe group treated with the medicated sera, and Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group transfected with lncRNA-NEAT1 overexpression vectors and treated with the medicated sera. qPCR, Western blot analysis, flow cytometry, and cell counting kit (CCK8) assay were performed to determine the mRNA and protein expression of lncRNA NEAT1, miR-485-5p and STAT3, and to evaluate cell proliferation and apoptosis. The two independent samples t-test was used for comparisons between two groups, one-way analysis of variance for comparisons among multiple groups, and least significant difference (LSD) t-test for multiple comparisons. Results:The IL-17-induced HaCaT cell group showed significantly increased relative expression levels of lncRNA NEAT1 and STAT3 mRNA (1.84 ± 0.21, 2.20 ± 0.24, respectively) and significantly increased protein expression of STAT3 and p-STAT3 (1.27 ± 0.13, 2.43 ± 0.16, respectively), but significantly decreased expression level of miR-485-5p (0.32 ± 0.04) compared with the NHEK group (lncRNA NEAT1 and STAT3 mRNA: 1.00 ± 0.11, 1.00 ± 0.11, respectively, both P < 0.05; STAT3 and p-STAT3 protein: 1.00 ± 0.11, 1.00 ± 0.10, t = 2.54, 3.02, respectively, both P < 0.05; miR-485-5p: 1.00 ± 0.12, t = 2.94, P = 0.015). FISH demonstrated that miR-485-5p and lncRNA NEAT1 were co-located in the cytoplasm of HaCaT cells. The double-luciferase reporter gene assay showed that the relative activity of luciferase was significantly lower in the miR-485-5p group than in the negative control group (both P < 0.05) after the transfection with wild-type lncRNA NEAT1 or STAT3 recombinant plasmids, while there were no significant differences between the miR-485-5p group and negative control group after the transfection with mutant lncRNA NEAT1 or STAT3 recombinant plasmids (both P > 0.05). Compared with the control group, the lncRNA-NEAT1 overexpression group showed significantly increased expression of lncRNA NEAT1 and STAT3 (including STAT3 mRNA, STAT3 protein, and p-STAT3 protein) in HaCaT cells (all P < 0.05), but significantly decreased miR-485-5p expression ( P < 0.05) ; the Xidi Liangxue recipe group showed significantly decreased expression of lncRNA NEAT1 and STAT3 (all P < 0.05), but significantly increased miR-485-5p expression compared with the control group ( P < 0.05) ; significantly decreased expression of lncRNA NEAT1 and STAT3, but significantly increased miR-485-5p expression was observed in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group compared with the lncRNA-NEAT1 overexpression group (all P < 0.05). After 24-, 48-, and 72-hour intervention, CCK8 assay showed that the proliferative activity of HaCaT cells was significantly higher in the lncRNA-NEAT1 overexpression group than in the control group (all P < 0.05), as well as in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group than in the Xidi Liangxue recipe group (all P < 0.05), and the cellular proliferative activity was significantly lower in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group and Xidi Liangxue recipe group than in the control group (all P < 0.05). The apoptosis rate was significantly lower in the lncRNA-NEAT1 overexpression group (5.84% ± 0.28%) than in the control group (14.75% ± 0.83%, LSD- t = 3.48, P = 0.002), but significantly higher in the Xidi Liangxue recipe group (35.72% ± 3.62%) than in the control group (LSD- t = 5.34, P = 0.001) ; the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group showed significantly increased apoptosis rate (27.64% ± 2.82%) compared with the lncRNA-NEAT1 overexpression group (LSD- t = 9.06, P < 0.001) . Conclusion:The Xidi Liangxue recipe could inhibit the proliferation of IL-17-induced HaCaT cells and promote their apoptosis, which may be related to the intervention in the lncRNA NEAT1/miR-485-5p/STAT3 regulatory network.
2.Bloodletting Acupuncture at Jing-Well Points on Hand Induced Autophagy to Alleviate Brain Injury in Acute Altitude Hypoxic Rats by Activating PINK1/Parkin Pathway.
Yong-Ping LI ; Meng-Xin LI ; Chao WANG ; Yun-di LI ; Yu-Ping SA ; Yi GUO
Chinese journal of integrative medicine 2023;29(10):932-940
OBJECTIVE:
To explore the protective effect of bloodletting acupuncture at twelve Jing-well points on hand (BAJP) on acute hypobaric hypoxia (AHH)-induced brain injury in rats and its possible mechanisms.
METHODS:
Seventy-five Sprague Dawley rats were divided into 5 groups by a random number table (n=15), including control, model, BAJP, BAJP+3-methyladenine (3-MA), and bloodletting acupuncture at non-acupoint (BANA, tail tip blooding) groups. After 7-day pre-treatment, AHH models were established using hypobaric oxygen chambers. The levels of S100B, glial fibrillary acidic protein (GFAP), superoxide dismutase (SOD), and malondialdehyde (MDA) in serum were measured by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling method were used to assess hippocampal histopathology and apoptosis. Transmission electron microscopy assay was used to observe mitochondrial damage and autophagosomes in hippocampal tissues. Flow cytometry was used to detect mitochondrial membrane potential (MMP). The mitochondrial respiratory chain complexes I, III and IV activities and ATPase in hippocampal tissue were evaluated, respectively. Western blot analysis was used to detect the protein expressions of Beclin1, autophagy protein 5 (ATG5), microtubule-associated protein 1 light chain 3 beta (LC3B), phosphatase and tensin homolog induced kinase 1 (PINK1), and Parkin in hippocampal tissues. The mRNA expressions of Beclin1, ATG5 and LC3-II were analyzed by quantitative real-time polymerase chain reaction.
RESULTS:
BAJP treatment reduced hippocampal tissue injury and inhibited hippocampal cell apoptosis in AHH rats. BAJP reduced oxidative stress by decreasing S100B, GFAP and MDA levels and increasing SOD level in the serum of AHH rats (P<0.05 or P<0.01). Then, BAJP increased MMP, the mitochondrial respiratory chain complexes I, III and IV activities, and the mitochondrial ATPase activity in AHH rats (all P<0.01). BAJP improved mitochondrial swelling and increased the autophagosome number in hippocampal tissue of AHH rats. Moreover, BAJP treatment increased the protein and mRNA expressions of Beclin1 and ATG5 and LC3-II/LC3-I ratio in AHH rats (all P<0.01) and activated the PINK1/Parkin pathway (P<0.01). Finally, 3-MA attenuated the therapeutic effect of BAJP on AHH rats (P<0.05 or P<0.01).
CONCLUSION
BAJP was an effective treatment for AHH-induced brain injury, and the mechanism might be through reducing hippocampal tissue injury via increasing the PINK1/Parkin pathway and enhancement of mitochondrial autophagy.
3.Series of group standards of Technical Specifications for Revision of Safety Information in Marketed Chinese Patent Medicine Instructions.
Bing ZHANG ; Xiao-Meng ZHANG ; Zhi-Jian LIN ; Ri-Na SA ; Jin-Tao LYU ; Hao WU ; Yao-Lei LI ; Hui-Zhe XU ; Zheng-Kai HUANG ; Yu-Bo GUO ; Xiang-Fei SU ; Xiao-Jiao DUAN
China Journal of Chinese Materia Medica 2022;47(2):285-294
Drug instructions,the statutory and technical documents recording effectiveness and safety information,are an important basis for guiding doctors,pharmacists,and patients to use drugs rationally,and their scientificity,standardization,and accuracy directly affect the medication safety of the public. The sections of adverse drug events,contraindications,precautions,warnings,and application for specific populations in drug instructions directly express safety information and measures for rational use of drugs. In the drug life cycle,marketing authorization holders( MAHs) need to update safety information in the instructions promptly to ensure the safety and effectiveness of clinical drug medication. At present,revising instructions is an important measure to control drug risks. In the drug life cycle,in order to standardize the revision of safety information in the instructions by MAHs and eliminate inexact terms such as " unclear",the Technical Specifications for Revision of Safety Information in Marketed Chinese Patent Medicine Instructions,a series of group standards,have been established under the guidance of Standardization Department,China Association of Chinese Medicine. Therefore,on the basis of the existing rules and regulations,the standardized technical procedures for revising instructions came into being to help clinical safe and rational medication of drugs,and implement the strategy of " Healthy China".
China
;
Drug-Related Side Effects and Adverse Reactions
;
Drugs, Chinese Herbal/adverse effects*
;
Humans
;
Medicine, Chinese Traditional
;
Nonprescription Drugs/adverse effects*
;
Reference Standards
4.Interpretation of Technical Specifications for Revision of Safety Information in Marketed Chinese Patent Medicine Instructions.
Xiao-Meng ZHANG ; Zhi-Jian LIN ; Bing ZHANG ; Ri-Na SA ; Jin-Tao LYU ; Hao WU ; Yao-Lei LI ; Hui-Zhe XU ; Zheng-Kai HUANG ; Yu-Bo GUO ; Xiang-Fei SU ; Xiao-Jiao DUAN
China Journal of Chinese Materia Medica 2022;47(2):295-300
Technical Specifications for Revision of Safety Information in Marketed Chinese Patent Medicine Instructions,a series of group standards,were proposed by Professor ZHANG Bing from Research Center for Pharmacovigilance and Rational Use of Traditional Chinese Medicine,and underwent centralized management by Chinese Association of Chinese Medicine. They were officially released on July 23 and implemented on July 31,2021. The series of group standards consist of six sections,including general principles,adverse drug events,contraindications,precautions,application for special populations,and warnings. The section of general principles is comprised of holistic and programmatic expressions,which explain the general technical requirements for revising the marketed Chinese patent medicine instructions. The other five sections focus on information collection,screening,transformation,and illustration of specific items,forming a standardized revision technical process. This series of standards is the result of multiple rounds of research and the suggestions of more than 200 experts in different professional fields of " medicine-pharmacy-management-law-enterprise" have been gathered therein to reach a consensus. With the purposes of establishing standardized technical specifications for the revision of safety information in the marketed Chinese patent medicine instructions,guiding marketing authorization holders in revising the instructions,filling the gaps in the research of Chinese patent medicine instructions,promoting the deve-lopment of pharmaceutical care and academic research,and encouraging the rational and safe medication of Chinese patent medicine,the series of group standards is of great significance.
China
;
Drug-Related Side Effects and Adverse Reactions
;
Drugs, Chinese Herbal/adverse effects*
;
Humans
;
Medicine, Chinese Traditional
;
Nonprescription Drugs/adverse effects*
;
Pharmacovigilance
5.Technical specification for Instructions for Clinical Application of Chinese Patent Medicines in China Association of Chinese Medicine.
Xiao-Meng ZHANG ; Ri-Na SA ; Bing ZHANG ; Dan ZHANG ; Jin-Tao LYU ; Xiao-Jiao DUAN ; Zhi-Jian LIN ; Yu-Bo GUO
China Journal of Chinese Materia Medica 2021;46(17):4563-4568
In order to solve the problems of confusion in clinical medication and imperfect instructions in Chinese patent medicines(CPMs), the Standardization Department of the China Association of Chinese Medicine and Center for Pharmacovigilance and Rational use of Chinese Medicine in Beijing University of Chinese Medicine jointly compiled the Instructions for Clinical Application of Chinese Patent Medicines(CPMs). As the interpretation and supplement of drug instruction information, it aims to guide clinical safety and rational use of CPMs. In addition, the technical specification for clinical application description of CPMs has been formulated, which covers the seven processes of "carding instructions, clinical investigation, data retrieval, data screening, evidence classification, path transformation and writing format". It will enable readers of Instructions for Clinical Application of Chinese Patent Medicines to understand the work behind the compilation.
Beijing
;
China
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Drugs, Chinese Herbal
;
Medicine, Chinese Traditional
;
Nonprescription Drugs
;
Pharmacovigilance
6.Analysis of trends on smoking prevalence and its risk factors in Shaanxi province 2007-2015
Weihua WANG ; Lin QIU ; Rina SA ; Zhiping HU ; Rong LIU ; Meng WU ; Feng LIU ; Tianhang ZHANG
Chinese Journal of Epidemiology 2020;41(3):395-399
Objective:To analyze the trend of smoking prevalence and its risk factors among adults in Shaanxi province from 2007 to 2015.Methods:We used data from China Chronic Disease and Risk Factor Surveillance in 2007, 2010, 2013 and 2015. The current smoking prevalence and trends of the four surveys were calculated. Its risk factors were analyzed by multivariate logistic regression from each survey and then from all pooled data of the three surveys.Results:The number of participants in 2007, 2010, 2013 and 2015 was 1 542, 3 000, 10 166 and 6 330, respectively. The current smoking prevalence dropped from 34.34 % in 2007 to 26.22 % in 2013, but increased to 28.33 % in 2015 (trend χ2 test: Z=2.53, P=0.01). The results from four pooled data showed that the current smoking prevalence of men was higher than that of women ( OR=75.03, 95 %CI: 63.57-88.55). The current smoking prevalence of people aged 45-59 was higher than that of people aged 18-44 ( OR=1.28, 95 %CI: 1.15-1.41). In addition, the current smoking prevalence of those who were educated for 7-9 years and more than 9 years were higher than those who were educated for less than 6 years (people with education for 7-9 years OR=1.44, 95 %CI: 1.29-1.61; people with education >9 years OR=1.43, 95 %CI: 1.26-1.63). The current smoking prevalence of the single was lower than those of married/cohabitants ( OR=0.54, 95 %CI: 0.37-0.77). The current smoking prevalence of retirees were lower than those of employees ( OR=0.46, 95 %CI: 0.38-0.57) and smoking prevalence of alcohol drinkers were higher than those of non-drinkers ( OR=2.92, 95 %CI: 2.67-3.19). Conclusion:From 2007 to 2015, the current smoking prevalence of Shaanxi population was high and the trends remained stable. It is necessary to strengthen smoking control and health education for men, people over 45 years old, people with education level 7 years and above, and working personnel in Shaanxi province.
7.Detection and analysis of clinical specimens of COVID-19 in Anhui province
Qingqing CHEN ; Jun HE ; Junling YUN ; Yonglin SHI ; Weiwei LI ; Yuan YUAN ; Yinglu GE ; Nan SA ; Meng WANG ; Zhuhui ZHANG ; Yong SUN
Chinese Journal of Experimental and Clinical Virology 2020;34(4):382-384
Objective:To examine 2019 novel coronavirus (2019-nCoV) RNA in clinical specimens of COVID-19 patients in Anhui province, and provide evidence for laboratory diagnosis of COVID-19 and risk assessment of clinical specimens.Methods:ORF1ab gene and N gene of 2019-nCoV were detected by real-time fluorescence RT-PCR in 466 clinical specimens of 197 COVID-19 cases. Chi-square test was used to analyze the differences in positive rates of specimens with clinical classification and time of onset.Results:The positive rates of 2019-nCoV in throat swab, sputum, serum, blood sample were 88.83%, 94.67%, 6.78% and 5.08%. The positive rate for 2019-nCoV RNA in throat swabs and sputum differed significantly ( χ2=8.994, P=0.003) in common cases during 7 days after illness onset. Conclusions:The positive rate of RNA in sputum was higher than throat swabs. 2019-nCoV RNA was detected in serum and blood specimens of COVID-19 cases. There was a risk of serum and blood specimens for transmission of COVID-19.
8. Analysis on chronic diseases and risk factors of adult residents in Shaanxi province
Weihua WANG ; Rina SA ; Lin QIU ; Zhiping HU ; Rong LIU ; Meng WU ; Feng LIU ; Shaonong DANG
Chinese Journal of Epidemiology 2019;40(12):1585-1589
Objective:
To explore the prevalence of chronic diseases and related risk factors in Shaanxi province.
Methods:
Multi-stage stratified cluster random sampling was used to collect the sample from permanent residents in 10 national surveillance points in Shaanxi province in 2015. Behavioral risk factors (smoking, drinking, diet and physical activity) were investigated by face-to-face interviews and biological risk factors (BMI, blood pressure, blood glucose and blood lipid) were collected by physical measurements and laboratory tests. Designed weight, no response weight and post hierarchical weight were taken into account in the data analysis. Binary logistic regression models were used to examine the pair-wise associations among 8 risk factors.
Results:
A total of 6 174 persons were included in the analysis. The following weighted prevalence were noticed in Shaanxi province in 2015, that including current smoking as 28.19
9.miR-1301/TRIAP1 Axis Participates in Epirubicin-Mediated Anti-Proliferation and Pro-Apoptosis in Osteosarcoma
Lijun YU ; Min MENG ; Yun BAO ; Chao ZHANG ; Bei GAO ; Rina SA ; Wenyuan LUO
Yonsei Medical Journal 2019;60(9):832-841
PURPOSE: Epirubicin is one of the most effective drugs against osteosarcoma. miR-1301 is involved in the occurrence and development of osteosarcoma. Whether miR-1301 is responsible for the chemosensitivity of osteosarcoma cells to epirubicin remains largely unknown. MATERIALS AND METHODS: U2OS and SAOS-2 cells were treated with various concentrations of epirubicin. Flow cytometry was employed to evaluate cell apoptotic rate. Cell proliferation was measured by Cell Counting Kit-8 assay. Western blot and quantitative real-time polymerase chain reaction were utilized to detect the expressions of B-cell lymphoma-2 (Bcl-2), Bcl-2 assaciated X protein (Bax), cleaved-caspase-3, cleaved-poly (ADP-ribose) polymerases (PARP1), TP53-regulated inhibitor of apoptosis 1 (TRIAP1), and microRNA-1301 (miR-1301). The relationship between miR-1301 and TRIAP1 was determined by luciferase reporter assay. RESULTS: Epirubicin inhibited proliferation in a dose-dependent manner, induced apoptosis, decreased the expression of Bcl-2, and increased the expressions of Bax, cleaved-caspase-3, and cleaved-PARP1 in osteosarcoma cells. miR-1301 was downregulated in U2OS and SAOS-2 cells. Importantly, epirubicin significantly increased the levels of miR-1301. Overexpression of miR-1301 suppressed proliferation and promoted apoptosis. Interestingly, those effects were enhanced by epirubicin. In contrast, miR-1301 depletion attenuated the epirubicin-mediated anti-osteosarcoma effect. miR-1301 negatively regulated the expression of TRIAP1 in U2OS and SAOS-2 cells. Furthermore, epirubicin inhibited the mRNA and protein levels of TRIAP1 by upregulating miR-1301 levels. Epirubicin suppressed cell proliferation by downregulating TRIAP1. CONCLUSION: miR-1301 was implicated in the chemosensitivity of osteosarcoma to epirubicin by modulating TRIAP1.
Apoptosis
;
B-Lymphocytes
;
Blotting, Western
;
Cell Count
;
Cell Proliferation
;
Epirubicin
;
Flow Cytometry
;
Luciferases
;
Osteosarcoma
;
Real-Time Polymerase Chain Reaction
;
RNA, Messenger
10.Cost-effectiveness Analysis of Xueshuantong for Injection versus Ginkgo Leaf Extract and Dipyridamole In-jection in the Treatment of Ischemic Stroke
Bei GAO ; Min MENG ; Jing YU ; Rina SA ; Yan YANG ; Bin GE ; Hongru ZHAO
China Pharmacy 2015;26(36):5105-5107
OBJECTIVE:To compare pharmacoeconomic and effect of Xueshuantong for injection and Ginkgo leaf extract and dipyridamole injection in the treatment of ischemic stroke. METHODS:Retrospective study was conducted. Totally 404 inpatients with ischemic stroke were divided into Xueshuantong group(271 cases)and ginkgo leaf extract and dipyridamole group(133 cas-es) according to clinical treatment programs. Based on the conventional treatment,patients in 2 groups were given Xueshuantong for injection and ginkgo leaf extract and dipyridamole injection,respectively. The average treatment course was 10 d. Cost-minimi-zation analysis was performed with the determination index of total effective rate. RESULTS:The total effective rates in Xueshuan-tong group and ginkgo leaf extract and dipyridamole group were 90.77% and 88.72%,respectively,the difference was not statisti-cally significant(P>0.05). The costs in 2 groups were 12 860.21 yuan and 13 155.40 yuan,respectively,and xueshuantong group had lower than ginkgo leaf extract and dipyridamde group. CONCLUSIONS:Both Xueshuantong for injection and Ginkgo leaf ex-tract and dipyridamole injection are effective in the treatment of ischemic stroke. However,the economy of Xueshuantong for injec-tion is superior to the other one.

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