1.Effects of IL-12 combined with GM-CSF on apoptosis of H22 hepatoma cells
Siqi LIU ; Qian ZHANG ; Xue SHAO ; Shengnan JIA ; Jingting MA ; Liulan PAN
Clinical Medicine of China 2017;33(7):591-595
Objective To investigate the effects of granulocyte macrophage colony-stimulating factor (GM-CSF) combined with interleukin-12 (IL-12) genes on apoptosis of hepatoma cells.Methods The hepatoma cell lines were cultured in vitro and were divided into four groups: GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group,negative control group (empty load group),respectively.The PIB-CMV3-GM-CSF and PIB-CMV3-IL-12 eukayotic expression vector was built,and 36 h after transfection,fluorescence microscope was used to detect the transfection effect;the expression level of IL-12,GM-CSF,p53,p38 and C-JUN mRNA were detected by RT-PCR,and Western blot was used to examine the expression level of IL-12,GM-CSF,p53,p38 and C-JUN protein.In addition,the flow cytometry was applied to detect cell apoptosis.Results Through fluorescence microscope,green fluorescence was observed in cells of GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group,indicating that the plasmid has successfully transferred into cells.In addition,the expression of p53mRNA in empty load group,GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group were 1.2±0.10,4.3±0.98,4.2±0.34,9.2±0.87,and the protein expression were 1.0±0.10,3.6±0.34,3.8±0.30,5.0±0.60.Compared with the empty load group,the expression level of p53 mRNA and protein were significantly increased in the three plasmid transfection groups (P<0.01).The expression of p53 mRNA and protein were significantly increased in co-transfection group than GM-CSF group and IL-12 group (P<0.01),while in the comparison with GM-CSF transfection group and IL-12 transfection group,the expression level of p53mRNA and protein in the co-transfection group could be improved to a higher degree(P<0.01).Meanwhile,p38 C-JUN mRNA expression levels in empty load group,GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group were as follows: 7.5± 0.9,3.5±0.45,3.7±0.25,1.0±0.11,while p38protein expression levels were 10.1±1.03,6.1± 0.67,7.1 ± 0.61,1.0 ± 0.12,respectively,C-JUN mRNA expression levels were 11.2 ± 1.20,4.1 ± 0.19,3.3 ± 0.30,1.0 ± 0.01,separately,C-JUN protein expression levels were 2.25 ± 0.2,1.8 ± 0.13,1.4 ± 0.12,1.0 ± 0.09.P38, C-JUN mRNA and protein levels were significantly reduced in the three plasmid transfection groups compared with the empty load group (P<0.01).The expression of p38,C-JUN mRNA and protein were reduced to a lower degree in co-transfection group than in GM-CSF transfection group and IL-12 transfection group (P<0.01).Flow cytometer showed that the hepatoma cell apoptosis rate of the empty load group,GM-CSF transfection group,IL-12 transfection group,co-transfection group were (3.43±0.9)%,(5.87±1.02)%,(7.32±1.1)%,(17.47±2.11)%,the rates of the three plasmid transfection groups were significantly higher than that of the empty load group (P<0.01).And the apoptosis rate was significantly increased in the co-transfected group compared with other plasmid groups (P<0.01). Conclusion The combination of GM-CSF and IL-12 could significantly accelerate the apoptosis of hepatoma cells by up-regulating the expression of p53,and down-regulating the expression of p38 and C-JUN.
2.Effects of Liuweidihuang pill on insulin levels in sera and pancreatic islets from spontaneous mouse models of human type 2 diabetes administrated with different doses
Lu Lü ; Yuanqiang ZHENG ; Jingcai ZHANG ; Siqi LI ; Xiaodong WANG ; Xing YIN ; Jia XU ; Xinrong HAN ; Yanchun SHI
Chinese Journal of Immunology 2016;32(3):350-353
Objective:To investigate the effects of Liuweidihuang pill on the insulin levels in sera and pancreatic islets from spontaneous mouse models of human type 2 diabetes administrated with different doses .Methods:The 6-8 week-old KK-Ay mice were randomly divided into three groups including no drug control group ,low-dose group and high-dose group,in addition C57BL/6J mice were used as a genetic control group .All the animals were given with different dose Liuweidihuang pill solutions or sterile distilled water by intragastrical administration for fifteen weeks .The fasting blood glucose ,body mass and food consumption were measured weekly .The serum insulin levels were surveyed by ELISA .And the insulin levels in the pancreas islets were detected by immunofluorescence and immunohistochemistry .Results:Decreased fasting blood glucose ,controlled body mass and food consumption ,and lower levels of insulin in the sera and pancreas islets were confirmed from the KK-Ay mice administered with Liuweidihuang pill .Furthermore,the low dose program exhibits a stronger effect .Conclusion:Liuweidihuang pill has exhibited relatively therapeutic effects in the spontaneous type 2 diabetes mice including controls of hyperglycemia and body mass and relieving insulin resistance .In addition , the low-dose regimen showed even better treatment in controlling insulin levels in the sera and pancreas islets .
3.The role of fluid biomarkers in the diagnosis of Alzheimer′s disease
Chinese Journal of Neurology 2023;56(3):245-250
The rapid advancement of biomarker research in Alzheimer′s disease (AD) has improved the accuracy of AD diagnosis, facilitated its early diagnosis and intervention, and has significant implications for disease-modifying therapy research and evaluation. Since its introduction, the AT(N) system has grown in importance in both the clinical and research aspects of AD. However, the fluid biomarkers in the AT(N) system do not yet adequately reflect the complex pathophysiology of AD. Other fluid biomarkers associated with neuroinflammation, synaptic dysfunction, and vascular dysfunction should be a valuable addition to the AT(N). In recent years, the development and application of peripheral blood biomarkers have greatly enriched the fluid biomarker system for AD and are expected to greatly facilitate early screening and diagnosis of the disease. Here, a brief overview of current research on AT(N) and other pathogenesis-related fluid biomarkers is presented, and the article proposes that fluid biomarkers for AD should be studied from a variety of pathophysiological perspectives in order to improve their role in disease diagnosis. This article also discusses the role and limitations of fluid biomarkers in the diagnosis of AD, as well as the future development and application of fluid biomarkers in AD.
4.Functional near infrared spectroscopy of the impact of childhood traumatic experiences on risky decisions in early adulthood
Chinese Journal of School Health 2024;45(10):1460-1464
Objective:
To investigate the impact of childhood traumatic experiences on individual risktaking decisions in early adulthood using functional nearinfrared spectroscopy (fNIRS), so as to provide the reference for clarifying the brain mechanisms underlying the impact of childhood trauma on individual risky decision.
Methods:
From December 2023 to March 2024, 28 children with childhood trauma experiences (trauma group) and 32 healthy college students (control group) were selected from Jining Medical University by a combination of stratified descent and convenient sampling methods. All subjects participated in the Iowa Game task fNIRS scanning. The brain activation, functional connectivity, graph theory properties (degree centrality, betweenness centrality, and local efficiency), and Receiver Operating Characteristic (ROC) analysis were performed by using preprocessing fNIRS data.
Results:
Compared with control group, trauma group showed significantly fewer choice times in the inferior deck (Z=-0.88), and showed significantly decreased activation levels in the right frontalpolar (Z=-2.59), as well as showed significant decreased functional connectivity between left dorsolateral prefrontal and in right dorsolateral prefrontal (Z=-3.78), and between left dorsolateral prefrontal cortex and the right frontal pole (Z=-3.68)(P<0.05). The central index of right inferior frontal gyrus in the trauma group was higher than that in the control group, while the central index of left and right dorsolateral frontal lobes was lower than that in the control group (Z=2.13, -2.53, -2.12, P<0.05). The centrality index of the right inferior frontal gyrus in the trauma group was higher than that in the control group (Z=2.47, P<0.05). The local efficiency indicators of the right inferior frontal gyrus, left and right frontal pole in the trauma group were higher than those in the control group (Z=2.51, 2.17, 2.53, P<0.05). The results of the ROC curve analysis showed that the local efficiency achieved the highest area under the curve (AUC=0.68).
Conclusions
Young adults with childhood trauma experience tend to choose lower loss, and the frontal pole shows a lack of activation in the whole process of risk decision performance. The abnormalities in the brain connectivity and network properties might be the neural basis of excessive defense mechanisms that childhood trauma leads to risky decisions.
5.The C-terminal portion of the nucleocapsid protein demonstrates SARS-CoV antigenicity.
Guozhen LIU ; Shaohui HU ; Yongwu HU ; Peng CHEN ; Jianning YIN ; Jie WEN ; Jingqiang WANG ; Liang LIN ; Jinxiu LIU ; Bo YOU ; Ye YIN ; Shuting LI ; Hao WANG ; Yan REN ; Jia JI ; Xiaoqian ZHAO ; Yongqiao SUN ; Xiaowei ZHANG ; Jianqiu FANG ; Jian WANG ; Siqi LIU ; Jun YU ; Heng ZHU ; Huanming YANG
Genomics, Proteomics & Bioinformatics 2003;1(3):193-197
In order to develop clinical diagnostic tools for rapid detection of the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) and to identify candidate proteins for vaccine development, the C-terminal portion of the nucleocapsid (NC) gene was amplified using RT-PCR from the SARS-CoV genome, cloned into a yeast expression vector (pEGH), and expressed as a glutathione S-transferase (GST) and Hisx6 double-tagged fusion protein under the control of an inducible promoter. Western analysis on the purified protein confirmed the expression and purification of the NC fusion proteins from yeast. To determine its antigenicity, the fusion protein was challenged with serum samples from SARS patients and normal controls. The NC fusion protein demonstrated high antigenicity with high specificity, and therefore, it should have great potential in designing clinical diagnostic tools and provide useful information for vaccine development.
Antigens, Viral
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immunology
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Cloning, Molecular
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Enzyme-Linked Immunosorbent Assay
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Genetic Vectors
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Genome, Viral
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Humans
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Nucleocapsid Proteins
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genetics
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immunology
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Recombinant Fusion Proteins
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genetics
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isolation & purification
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metabolism
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SARS Virus
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genetics
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immunology
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Yeasts
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genetics
6.A Preliminary Proteomic Study of Peitu Qingxin Granules for the Treatment of Atopic Dermatitis
Junfeng LIU ; Kai ZHANG ; Xiumei MO ; Saiya WANG ; Fenggen YAN ; Siqi YE ; Jinjing JIA ; Yu ZHANG ; Dacan CHEN ; Xiaobo YU
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(2):191-199
Objective A serum proteomic approach was used to explore the targets of action of Peitu Qingxin Granules(composed of Rhizoma Atractylodis Macrocephalae,Forsythiae Fructus,Imperatae Rhizoma,Pseudostellariae Radix,etc.)in the treatment of atopic dermatitis.Methods Five patients with atopic dermatitis were selected and treated with Peitu Qingxin Granules for 12 weeks,and five healthy volunteers were used as controls.The clinical core evaluation indexes of atopic dermatitis patients after treatment,including Eczema Area and Severity Index/Scoring Atopic Dermatitis(EASI/SCORAD),Pruritus Score,Patient-Oriented Eczema Measure(POEM),and quality of life index,were assessed.Serum samples were examined using data-independent acquisition-mass spectrometry(DIA-MS)technology,and serum differential proteins between atopic dermatitis patients and healthy people,as well as serum differential proteins in atopic dermatitis patients before and after treatment with Peitu Qingxin Granules were screened according to P<0.05 and Fold Change>1.2.GO function enrichment analysis and KEGG pathway enrichment analysis were performed on the differential proteins.Results(1)Compared with the pre-treatment period,the clinical core evaluation indexes of patients with atopic dermatitis,including the EASI/SCORAD,Pruritus Score,POEM,and quality-of-life index,were significantly improved after treatment,and the differences were all statistically significant(P<0.05,P<0.01).(2)A total of 28 differential proteins were analyzed in the healthy control group and atopic dermatitis group,of which 12 proteins expressions were increased and 16 proteins were decreased,including ALAD(δ-aminolevulinic acid dehydrogenase),LTA4H(leukotriene A-4 hydrolase),CA1(carbonic anhydrase 1),F11(coagulation factor XI),and LCP1(lymphocyte cytoplasmic protein 1),etc..The main signaling pathways involved are PI3K-AKT signaling pathway,lipids and atherosclerosis,ECM-receptor interaction,platelet activation,NF-κB signaling pathway,and neutrophil extracellular trap formation.(3)A total of 12 different proteins were analyzed in atopic dermatitis patients before and after treatment with Peitu Qingxin Granules,of which 8 proteins were increased and 4 proteins were decreased,including ALAD,FGA(fibrinogen α-chain),IGHV3-64D,and IGHV3-38.They were mainly involved in signaling pathways such as lipids and atherosclerosis,complement pathway,Staphylococcus aureus infection,NF-κB signaling pathway,fluid shear stress and atherosclerosis.(4)The expressions of three protein targets including ALAD,FGA and IGHV3-64D,were significantly down-regulated in patients with atopic dermatitis and significantly up-regulated after treatment with Peitu Qingxin Granules.Conclusion The differentially expressed proteins ALAD,FGA and IGHV3-64D may be the action targets of Peitu Qingxin Granules in the treatment of atopic dermatitis,which lays the foundation for further experimental validation.
7.Detection and evaluation of the signals of amlodipine and lercanidipine based on FAERS database
Guizun ZHONG ; Ni ZHANG ; Hongli WANG ; Siqi CHEN ; Li GONG ; Lingyun PAN ; Yuntao JIA
China Pharmacy 2022;33(21):2647-2653
OBJECTIVE To detect and evaluate the signals of amlodipine and lercanidipine -induced adverse drug events (ADE). METHODS All ADE reports about “amlodipine”and“lercanidipine”were searched from FAERS database during Jan . 1st,2004 to Sept . 30th,2021. Reported odds ratio and Bayesian confidence propagation neural network were used to detect ADE signals. The moderately strong signals and strong signals in key systems were selected for analysis . RESULTS From FAERS database,249 657 and 10 558 reports were extracted with amlodipine and lercanidipine as suspect drugs ,respectively. In this study,62 and 58 signals related to amlodipine and lercanidipine were detected respectively . At the same time ,moderately strong signals of peripheral edema , hypotension, orthostatic hypotension and hypovolemic shock were detected in the twodrugs,all of which were common adverse reactions of the two drugs. The special ADEs detected in this study were as follows: in the respiratory system , chest and mediastinaldisease system ,strong signals of non -cardiogenic pulmonary edema were detected for amlodipine ,and strong signals of dyspnea at rest for lercanidipine ;in gastrointestinal diseases ,strong signals of gingival hypertrophy were detected only for amlodipine;in skin and subcutaneous tissue disease system ,moderately strong signals related to “vasculitis”were detected for both drugs,moderately strong signals related to linear IgA disease were detected for amlodipine ,and moderately strong signals related to bullous dermatitis were detected for lercanidipine ;in the renal and urinary system disease system ,the signals of acute renal injury were detected for both drugs (amlodipine was detected as a moderately strong signal ,and lecardipine was detected as a strong signal ); in the mental system ,moderately strong signals related to suicide were detected for amlodipine . Both hypotension and acute renal injury ranked in the top two in the number of reports of the two drugs . The time scan results of the information component (IC)of this study showed that the IC values of non -cardiogenic pulmonary edema and suicide completion signals of amlodipine increased from 0.76,-0.49 to 4.48 and 1.95 respectively,and the confidence intervals narrowed from (-0.44,1.97),(-1.01,0.03)to (4.24,4.72)and(1.90,2.01)respectively during 2004 to 2021,suggesting that the signals kept stable . CONCLUSIONS The risks of peripheral edema ,hypotension,arrhythmia,pulmonary edema ,gingival hyperplasia ,skin related ADE ,acute renal injury , depression and suicide should be alert when using amlodipine and lercanidipine in clinic .
8.Complete genome sequences of the SARS-CoV: the BJ Group (Isolates BJ01-BJ04).
Shengli BI ; E'de QIN ; Zuyuan XU ; Wei LI ; Jing WANG ; Yongwu HU ; Yong LIU ; Shumin DUAN ; Jianfei HU ; Yujun HAN ; Jing XU ; Yan LI ; Yao YI ; Yongdong ZHOU ; Wei LIN ; Hong XU ; Ruan LI ; Zizhang ZHANG ; Haiyan SUN ; Jingui ZHU ; Man YU ; Baochang FAN ; Qingfa WU ; Wei LIN ; Lin TANG ; Baoan YANG ; Guoqing LI ; Wenming PENG ; Wenjie LI ; Tao JIANG ; Yajun DENG ; Bohua LIU ; Jianping SHI ; Yongqiang DENG ; Wei WEI ; Hong LIU ; Zongzhong TONG ; Feng ZHANG ; Yu ZHANG ; Cui'e WANG ; Yuquan LI ; Jia YE ; Yonghua GAN ; Jia JI ; Xiaoyu LI ; Xiangjun TIAN ; Fushuang LU ; Gang TAN ; Ruifu YANG ; Bin LIU ; Siqi LIU ; Songgang LI ; Jun WANG ; Jian WANG ; Wuchun CAO ; Jun YU ; Xiaoping DONG ; Huanming YANG
Genomics, Proteomics & Bioinformatics 2003;1(3):180-192
Beijing has been one of the epicenters attacked most severely by the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) since the first patient was diagnosed in one of the city's hospitals. We now report complete genome sequences of the BJ Group, including four isolates (Isolates BJ01, BJ02, BJ03, and BJ04) of the SARS-CoV. It is remarkable that all members of the BJ Group share a common haplotype, consisting of seven loci that differentiate the group from other isolates published to date. Among 42 substitutions uniquely identified from the BJ group, 32 are non-synonymous changes at the amino acid level. Rooted phylogenetic trees, proposed on the basis of haplotypes and other sequence variations of SARS-CoV isolates from Canada, USA, Singapore, and China, gave rise to different paradigms but positioned the BJ Group, together with the newly discovered GD01 (GD-Ins29) in the same clade, followed by the H-U Group (from Hong Kong to USA) and the H-T Group (from Hong Kong to Toronto), leaving the SP Group (Singapore) more distant. This result appears to suggest a possible transmission path from Guangdong to Beijing/Hong Kong, then to other countries and regions.
Genome, Viral
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Haplotypes
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Humans
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Mutation
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Open Reading Frames
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Phylogeny
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SARS Virus
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genetics
9.A genome sequence of novel SARS-CoV isolates: the genotype, GD-Ins29, leads to a hypothesis of viral transmission in South China.
E'de QIN ; Xionglei HE ; Wei TIAN ; Yong LIU ; Wei LI ; Jie WEN ; Jingqiang WANG ; Baochang FAN ; Qingfa WU ; Guohui CHANG ; Wuchun CAO ; Zuyuan XU ; Ruifu YANG ; Jing WANG ; Man YU ; Yan LI ; Jing XU ; Bingyin SI ; Yongwu HU ; Wenming PENG ; Lin TANG ; Tao JIANG ; Jianping SHI ; Jia JI ; Yu ZHANG ; Jia YE ; Cui'e WANG ; Yujun HAN ; Jun ZHOU ; Yajun DENG ; Xiaoyu LI ; Jianfei HU ; Caiping WANG ; Chunxia YAN ; Qingrun ZHANG ; Jingyue BAO ; Guoqing LI ; Weijun CHEN ; Lin FANG ; Changfeng LI ; Meng LEI ; Dawei LI ; Wei TONG ; Xiangjun TIAN ; Jin WANG ; Bo ZHANG ; Haiqing ZHANG ; Yilin ZHANG ; Hui ZHAO ; Xiaowei ZHANG ; Shuangli LI ; Xiaojie CHENG ; Xiuqing ZHANG ; Bin LIU ; Changqing ZENG ; Songgang LI ; Xuehai TAN ; Siqi LIU ; Wei DONG ; Jun WANG ; Gane Ka-Shu WONG ; Jun YU ; Jian WANG ; Qingyu ZHU ; Huanming YANG
Genomics, Proteomics & Bioinformatics 2003;1(2):101-107
We report a complete genomic sequence of rare isolates (minor genotype) of the SARS-CoV from SARS patients in Guangdong, China, where the first few cases emerged. The most striking discovery from the isolate is an extra 29-nucleotide sequence located at the nucleotide positions between 27,863 and 27,864 (referred to the complete sequence of BJ01) within an overlapped region composed of BGI-PUP5 (BGI-postulated uncharacterized protein 5) and BGI-PUP6 upstream of the N (nucleocapsid) protein. The discovery of this minor genotype, GD-Ins29, suggests a significant genetic event and differentiates it from the previously reported genotype, the dominant form among all sequenced SARS-CoV isolates. A 17-nt segment of this extra sequence is identical to a segment of the same size in two human mRNA sequences that may interfere with viral replication and transcription in the cytosol of the infected cells. It provides a new avenue for the exploration of the virus-host interaction in viral evolution, host pathogenesis, and vaccine development.
Base Sequence
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China
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Cluster Analysis
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Gene Components
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Genetic Variation
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Genome, Viral
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Genotype
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Molecular Sequence Data
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Phylogeny
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Reverse Transcriptase Polymerase Chain Reaction
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SARS Virus
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genetics
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Sequence Analysis, DNA
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Severe Acute Respiratory Syndrome
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genetics
10.Historical Evolution and Textual Research on Classical Prescription Taohe Chengqitang
Jinbao WANG ; Lei ZHANG ; Lin TONG ; Siqi JIA ; Zihan JIA ; Danping ZHENG ; Bing LI ; Huamin ZHANG ; Sihong LIU ;
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(18):135-143
Taohe Chengqitang is a classical prescription published in The Catalogue of Ancient Classic Recipes (The First Batch). This study systematically summarized traditional Chinese medicine (TCM) ancient books and modern clinical reports on Taohe Chengqitang and investigated its origin, composition, basis, and historical evolution in processing requirements, dosage, preparation and administration, and functions. The findings indicated different opinions on drug basis, preparation of decoction pieces, and drug dosage conversion. Based on sufficient literature review, the actual development needs of classical prescriptions, and the usage habits of modern clinical practice, this study recommended dried mature seeds of Prunus persica or P. davidiana for Persicae Semen, dried twigs of Cinnamomum cassia for Cinnamomi Ramulus, honey-fried Glycyrrhiza uralensis for Glycyrrhizae Radix et Rhizoma, raw Rheum palmatum, R. tanguticum, or R. offìcinale for Rhei Radix et Rhizoma, and mirabilite for Natrii Sulfas. In terms of drug dosage, the doses directed converted from weights and measures in the Han dynasty were large. According to the common doses in modern clinical practice, conversion was carried out based on 6 g of Cinnamomi Ramulus, 6 g of Natrii Sulfas, 6 g of Glycyrrhizae Radix et Rhizoma, and 12 g of Rhei Radix et Rhizoma. The weight of Persicae Semen should be determined according to the actual measurement. In terms of preparation and administration, the drugs in the pot were decocted with 1 400 mL of water to obtain 500 mL of decoction, which was filtered and slightly boiled in the presence of Natrii Sulfas. About 100 mL of warm decoction was taken before meals, three times a day. It was recorded in the ancient books that Taohe Chengqitang was indicated for the syndromes of heat invading the bladder and accumulating inside in the case of released exterior syndrome in Taiyang. After symptom-based flexible application by doctors, it is currently commonly used in the treatment of diabetic nephropathy, constipation, chronic renal failure, and other diseases.