1.Comparison of Protein and Polypeptide Components and Antithrombotic Activity In Vitro of Three Preparations Containing Hirudo
Wanling ZHONG ; Yunnan MA ; Jinhong YE ; Xiaoyu FAN ; Huijuan SHEN ; Rui YUAN ; Yaxuan ZHANG ; Zhuyuan LIU ; Shouying DU ; Pengyue LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):184-194
		                        		
		                        			
		                        			ObjectiveTo compare the contents and relative molecular weight distributions of proteins and polypeptides in Naoxuekang dropping pills, Huoxue Tongmai capsules and Maixuekang capsules of Hirudo single medicinal preparations, to evaluate the in vitro anticoagulant, antiplatelet and fibrinolytic activities of the three preparations, and to investigate the effects of temperature, pH and digestive enzymes on the anticoagulant activities of the three preparations. MethodsThe contents of soluble proteins and polypeptides in the three preparations were determined by bicinchoninic acid assay(BCA) and Bradford method, and the relative molecular weight distributions of the three preparations were determined by electrophoresis combined with gel chromatography. The antithrombin activity of the three preparations was evaluated by fibrinogen-thrombin time(Fibg-TT) method, and their anticoagulant activities were further assessed by the elongations of activated partial thromboplastin time(APTT), prothrombin time(PT) and thrombin time(TT). The antiplatelet aggregation activities of the three preparations were measured by turbidimetry and the fibrinolytic activities were measured by fibrin plate method. Relative TT was used as index to investigate the effects of temperature, pH and digestive enzyme buffer on anticoagulant activities of the three preparations. ResultsAt the lowest single dosage, the contents of proteins and polypeptides were in the order of Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. Both Huoxue Tongmai capsules and Maixuekang capsules had 11 electrophoretic bands between 4.0 kDa and 90 kDa, the bands of Maixuekang capsules were more clear in the range of >25 kDa, and there was 1 obvious band at 14 kDa for the two capsules. Huoxue Tongmai capsules had one specific band at 9.0 kDa and Maixuekang capsules had one specific band at 48.0 kDa. Naoxuekang dropping pills only had 2 electrophoretic bands at 6.5 kDa and 8.5 kDa, primarily containing peptides below 2 kDa, most of which were oligopeptides. The anticoagulant activity concentrations of the three preparations exhibited a certain dose-dependent effect. At the lowest single dosage, The anticoagulant activity concentrations were ranked as Naoxuekang dropping pills>Huoxue Tongmai capsules>Maixuekang capsules. The prolongation effect of the three preparations on coagulation time was dose-dependent. At the same concentration, the prolongation effect of Naoxuekang dropping pills and Huoxue Tongmai capsules was APTT prolongation rate>TT prolongation rate>PT prolongation rate, whereas for Maixuekang capsules, the sequence was TT prolongation rate>APTT prolongation rate>PT lengthening rate. At the single minimum dosage, the order of APTT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules≈Naoxuekang dropping pills, the order of PT prolongation rate was Naoxuekang dropping pills≈Maixuekang capsules>Huoxue Tongmai capsules, and the order of TT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. The three preparations showed dose-dependent effects on platelet aggregation induced by adenosine diphosphate(ADP) and arachidonic acid(AA), and the effect induced by ADP was stronger than that induced by AA. The anti-platelet aggregation effect of Naoxuekang dropping pills was significantly stronger than that of Maixuekang capsules(P<0.01), whereas Huoxue Tongmai capsules had the effect of promoting platelet aggregation. None of the three preparations had the ability to dissolve fibrin. The anticoagulant activity of Naoxuekang dropping pills was least affected by heating, while the activities of the two capsules decreased significantly within 5 min above 80 ℃, and continued to decrease within 2 h. Compared with pure water, the anticoagulant activities of the three preparations could be increased by 1-3 times under strong acidity(pH 1-3). In the pepsin buffer, the anticoagulant activity of Naoxuekang dropping pills could be increased by 1-3 times, while the anticoagulant activities of Huoxue Tongmai capsules and Maxuekang capsules were significantly decreased, the lowest levels were about 60% and 20%, respectively. In trypsin buffer, the anticoagulant activities of Naoxuekang dropping pills, Huoxue Tongmai capsules and Maixuekang capsules decreased significantly, and the lowest levels decreased to about 41%, 41% and 35%, respectively. ConclusionThe contents of proteins and polypeptides and relative molecular weights of the preparations derived from lyophilized fresh Hirudo powder, dried Hirudo powder and reflux extract of Hirudo decrease sequentially, and the anticoagulant activity decrease gradually, but the anticoagulant pathway is different. And the anti-platelet aggregation activity of the reflux extract is significantly enhanced. The heat resistance and gastrointestinal stability of the three preparations increase successively, and the first two are suitable for enteric-soluble preparations, while the latter is suitable for routine oral administration. The above results can provide data reference for the rationality of different preparation methods, active substances, pharmacodynamics and mechanism of Hirudo preparations. 
		                        		
		                        		
		                        		
		                        	
		                				2.Cloning and gene functional analysis study of dynamin-related protein GeDRP1E  gene in Gastrodia elata 
		                			
		                			Xin FAN ; Jian-hao ZHAO ; Yu-chao CHEN ; Zhong-yi HUA ; Tian-rui LIU ; Yu-yang ZHAO ; Yuan YUAN
Acta Pharmaceutica Sinica 2024;59(2):482-488
		                        		
		                        			
		                        			 The gene 
		                        		
		                        	
3.Scholars'consensus on the construction and development of chinese medical humanities:summary of"seminar on the construction of Chinese medical humanities"held in Harbin in August 2023
Jinfan WANG ; Mei YIN ; Yue WANG ; Huan LIU ; Zhong HE ; Yunzhang LIU ; Rui DENG ; Min CHEN ; Junrong LIU ; Yongfu CAO ; Donghong WANG ; Hongjiang ZHANG ; Fengxiang LU ; Yu CHENG ; Yuan HE ; Fang SHAN
Chinese Medical Ethics 2024;37(2):248-252
		                        		
		                        			
		                        			On August 2-4,2023,the"Third Summit Forum on'Building a Community of Shared Future for Doctors and Patients'"was jointly organized by institutions such as the Chinese Medical Ethics,the Hospital Humanities Management and Talent Training Special Committee of the China Population and Culture Promotion Association,Center for Ethical Studies of Renmin University of China,the Newspaper for China's Physicians,the China Health Law Society,the China Anti-Cancer Association,and the China Association For Ethical Studies in Harbin.The conference arranged a sub-forum for the"Seminar on the Construction of Chinese Medical Humanities",with domestic medical humanities scholars attending the conference.After heated discussions at the seminar,the Scholars'Consensus on the Construction and Development of Chinese Medical Humanities was formed.It was proposed that in the new era,it is urgent to build the medical humanities discipline,as well as lead the academic integration and development of medical humanities under the core socialist values.At the same time,for the construction of the medical humanities discipline,it is necessary to optimize the organizational mechanism,prosper and develop the overall framework of the medical humanities discipline,accelerate the construction of a professional teaching team for the medical humanities discipline,promote the establishment of a new carrier medical humanities education and teaching in cultivating morality and nurturing talents,as well as focus on solving problems related to the cultivation of medical humanities graduate students.
		                        		
		                        		
		                        		
		                        	
5.The taste correction process of ibuprofen oral solution based on the combination of electronic tongue technology and artificial taste comprehensive evaluation
Rui YUAN ; Yun-ping QU ; Yan WANG ; Ya-xuan ZHANG ; Wan-ling ZHONG ; Xiao-yu FAN ; Hui-juan SHEN ; Yun-nan MA ; Jin-hong YE ; Jie BAI ; Shou-ying DU
Acta Pharmaceutica Sinica 2024;59(8):2404-2411
		                        		
		                        			
		                        			 This experiment aims to study the taste-masking effects of different kinds of corrigent used individually and in combination on ibuprofen oral solution, in order to optimize the taste-masking formulation. Firstly, a wide range of corrigent and the mass fractions were extensively screened using electronic tongue technology. Subsequently, a combination of sensory evaluation, analytic hierarchy process (AHP)-fuzzy mathematics evaluation, and Box-Behnken experimental design were employed to comprehensively assess the taste-masking effects of different combinations of corrigent on ibuprofen oral solution, optimize the taste-masking formulation, and validate the results. The study received ethical approval from the Review Committee of the Beijing University of Chinese Medicine (ethical code: 2024BZYLL0102). The results showed that corrigent fractions and types were screened separately through single-factor experiments. Subsequently, a Box-Behnken response surface design combined with AHP and fuzzy mathematics evaluation was used to fit a functional model: 
		                        		
		                        	
6.Study on the Value of Serum miR-145 and P53 Antibody Detection in the Evaluation of Therapeutic Effect and Prognosis in the Treatment of Breast Cancer with Long Bei Xiaoyao Powder
Yuan-Kai LYU ; Rui YIN ; Qing-Zhong WEI ; Yi REN ; Ya-Hui ZHANG ; Feng-Qin SHI
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(11):2869-2875
		                        		
		                        			
		                        			Objective To explore the value of serum microRNAs-145(miR-145)and P53 antibody detection in the evaluation of therapeutic effect and prognosis in the treatment of breast cancer with Long Bei Xiaoyao Powder.Methods Ninety cases of breast cancer patients admitted to Beijing Aerospace General Hospital from January 2020 to January 2021 were selected as the study subjects.The patients were randomly divided into an observation group and a control group by random number table method,with 45 cases in each group.All of the patients in the two groups received individualized comprehensive treatment,and additionally the observation group was treated with Long Bei Xiaoyao Powder.The course of treatment covered three months.Real-time fluorescence quantitative polymerase chain reaction(PCR)method was used to detect serum miR-145 expression level,and enzyme-linked immunosorbent assay(ELISA)was used to detect serum P53 antibody expression level.The differences in the pre-and post-treatment expression levels of serum miR-145 and P53 antibodies in the two groups were compared,the relationship between the serum miR-145 and P53 antibody expression level and the poor prognosis of the patients in the observation group was explored by logistic regression analysis,and receiver operating characteristic curve(ROC)was drawn to analyze the value of the serum miR-145 and P53 antibody detection in the prediction of poor prognosis in the observation group.Results(1)After treatment,the detection value of serum miR-145 in the two groups of patients showed an increasing trend(P<0.05),while that of the P53 antibody showed a decreasing trend(P<0.05).The intergroup comparison showed that the detection value of serum miR-145 in the observation group was significantly higher than that of the control group,while the detection value of P53 antibody was significantly lower than that of the control group,the differences being all statistically significant(P<0.05).(2)In the observation group,the detection value of serum miR-145 in the patients with good prognosis was significantly higher than that in the patients with poor prognosis,and the detection value of serum P53 antibody in the patients with good prognosis was significantly lower than that in the patients with poor prognosis,the differences being all statistically significant(P<0.05).(3)Logistic regression analysis showed that high expression of miR-145 was a protective factor for the patients with poor prognosis in the observation group,and high expression of P53 antibody was a risk factor for the patients with poor prognosis in the observation group(P<0.05).(4)The area under the ROC curve(AUC)for the prediction of prognosis of the patients in the observation group by the combination of serum miR-145 and P53 antibody was the largest(0.877,95%CI:0.769-0.985),and its sensitivity and specificity were 93.75%and 93.10%,respectively,with statistically significant differences(P<0.05).Conclusion Low expression of serum miR-145 and high expression of serum P53 antibody are presented in the breast cancer patients with poor prognosis,and Long Bei Xiaoyao Powder exerts satisfactory therapeutic effect in treating breast cancer,which is conducive to enhancing the expression level of serum miR-145 and reducing the expression level of serum P53 antibody.
		                        		
		                        		
		                        		
		                        	
7.Fourth investigation and analysis of the quality control situation in the critical care medicine of traditional Chinese medicine hospitals in Sichuan province
Jun CHEN ; Xingyue CHEN ; Kunlan LONG ; Rui YUAN ; Song ZHANG ; Xiaobin LI ; Xingmei ZHONG ; Kaichen ZHANG ; Peng DING ; Peiyang GAO
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2024;31(4):459-464
		                        		
		                        			
		                        			Objective To enhance the quality of medical services in the intensive care unit(ICU)of provincial traditional Chinese medicine(TCM)hospitals.Methods In November 2023,Sichuan Provincial Critical Care Medicine Quality Control Center of TCM launched the"quality control supervision project scoring standard for critical care medicine of TCM"to conduct quality control evaluation and business guidance for all TCM hospitals with independent ICU.The survey covered structural indicators,control indicators,participation of TCM,development of new technologies,and diagnosis and treatment programs for dominant diseases.Results In terms of structural indicators:a total of 110 TCM hospitals in the province have independent ICU,an increase of 1.12 times compared with 2019.The control indicators showed that the ICU patients admission rate was higher than that of the national ICU admission rate in 2017,and the admission rate of patients with acute physiology and chronic health evaluation Ⅱ(APACHEⅡ)score≥15 points increased.However,the mortality of ICU exceeded the national average.The implementation of core indicators had been significantly improved,but the incidence of outcome indicators such as ICU ventilator-associated pneumonia(VAP),ICU intravascular catheter-related bloodstream infection(CRBSI),and ICU catheter-related urinary tract infection(CAUTI)had increased since 2019,mainly in secondary hospitals.The average number of new technologies was about(5.5±3.4),the participation rate of TCM decreased,and the dominant diseases increased compared with 2019,mainly sepsis,respiratory failure and hemorrhagic stroke.Conclusions The number of ICU units in TCM hospitals at all levels in Sichuan province has grown rapidly,and key performance indicators have also improved compared to previous periods.However,greater efforts are still needed in preventing the occurrence rates of VAP,CRBSI,and CAUTI.There is a shortage of medical resources allocation,and the imbalance in regional medical resources and professional training remains an urgent issue to be addressed.Additionally,the participation rate of TCM and the dominant diseases need further enhancement.
		                        		
		                        		
		                        		
		                        	
8.Investigation of tick species and potential pathogenic ricks in certain areas of Wuwei City,Gansu Province
Rui-Shan LI ; Zhen HE ; Xiang YUAN ; Shi-Wei SUN ; Yi-Wen LIU ; Wen-Kai ZHANG ; Lin ZHANG ; Yu-Hua WANG ; Zhen-Hua LU ; Zhao-Hua JI ; Zhong-Jun SHAO
Chinese Journal of Zoonoses 2024;40(4):328-333
		                        		
		                        			
		                        			To understand the distribution of ticks in the Wuwei Region,enrich tick species data,and provide a basis for the prevention of tick-borne diseases,tick were collected using flagging and tick-picking methods during the highest activity period from April to September in 2021 and 2022 in the mountainous areas of Wuwei City.The ticks were identified based on morpho-logical and molecular biological characteristics,and characteristic sequences were obtained.A systematic evolutionary tree was constructed using the neighbor-joining method in MEGA 11.0 software.In total,7 342 ticks collected in Wuwei,which be-longed to 5 species from 4 genera with in the Ixodidae family,which included Dermacentor nuttalli,Hyalomma asiaticum,Ixodes canisuga,Haemaphysalis longicornis and Haema-physalis danieli.Ticks of the same species clustered together into the same branch of an evolutionary tree.In the Wuwei Re-gion,five common tick species are found across various habi-tats,with each habitat featuring different distributions of tick species and populations.The Dermacentor nuttalli is the dom-inant tick species in this area.
		                        		
		                        		
		                        		
		                        	
9.Effect mechanism investigation of herb-partitioned moxibustion on relieving colon inflammation in Crohn disease rats based on neutrophil extracellular traps
Chi LU ; Jing XU ; Yuan LU ; Luyi WU ; Chunhui BAO ; Zhe MA ; Rui ZHONG ; Zhaoqin WANG ; Kexin SUN ; Handan ZHENG ; Zhijun WENG ; Yan HUANG ; Huangan WU ; Yanping YANG
Journal of Acupuncture and Tuina Science 2024;22(3):173-183
		                        		
		                        			
		                        			Objective:To explore the mechanism of herb-partitioned moxibustion in relieving rat intestinal inflammation by focusing on the neutrophil extracellular traps(NETs)in Crohn disease(CD)development. Methods:Rats were randomly divided into a normal group,a model group,a herb-partitioned moxibustion group,and a mesalazine group.The CD rat model was prepared with 2,4,6-trinitrobenzene sulfonic acid except for rats in the normal group.Rats in the normal group and model group did not receive any treatment but had the same fixation as the other groups.Rats in the herb-partitioned moxibustion group received herb-partitioned moxibustion at Qihai(CV6)and bilateral Tianshu(ST25).Rats in the mesalazine group received intragastric administration of mesalazine enteric-coated tablets.The general situation of rats in each group was recorded,and the histopathological changes in the colon were observed and scored by hematoxylin-eosin staining.The serum concentrations of NETs DNA(NETs-DNA),neutrophil elastase(NE)-DNA,and myeloperoxidase(MPO)-DNA were detected by ABC enzyme-linked immunosorbent assay,and the citrullinated histone 3(citH3),MPO,and NE protein and mRNA expression levels in rat colon tissue were observed by immunofluorescence and real-time quantitative polymerase chain reaction. Results:Compared with the normal group,the mucosal ulcer reached the muscularis,the epithelium was incomplete,the goblet cells decreased obviously with significant inflammatory cell infiltration in the colon;the colonic mucosa damage index(CMDI)score increased significantly(P<0.01);the serum NETs-DNA,NE-DNA,and MPO-DNA concentrations increased(P<0.05);the NE,citH3,and MPO protein and mRNA expression in the colonic tissue increased significantly in the model group(P<0.01 or P<0.05).Compared with the model group,the mucosal epithelium in the herb-partitioned moxibustion group and the mesalazine group was repaired and the goblet cells increased with a few infiltrating inflammatory cells in the colon;the CMDI score decreased(P<0.01);the serum NETs-DNA,NE-DNA,and MPO-DNA concentrations decreased(P<0.05);the NE,citH3,and MPO protein and mRNA expression in the colonic tissue was down-regulated(P<0.01 or P<0.05). Conclusion:Herb-partitioned moxibustion reduced the serum NETs complex and inhibited the protein and mRNA expression of NETs complex in the colon tissue,which may be one mechanism of herb-partitioned moxibustion in relieving colon mucosal inflammation in CD.
		                        		
		                        		
		                        		
		                        	
10.Cloning and gene function of dicarboxylate-tricarboxylate carrier protein in Gastrodia elata.
Jian-Hao ZHAO ; Yu-Chao CHEN ; Zhong-Yi HUA ; Tian-Rui LIU ; Yu-Yang ZHAO ; Lu-Qi HUANG ; Yuan YUAN
China Journal of Chinese Materia Medica 2023;48(12):3140-3148
		                        		
		                        			
		                        			The gene GeDTC encoding the dicarboxylate-tricarboxylate carrier protein in Gastrodia elata was cloned by specific primers which were designed based on the transcriptome data of G. elata. Bioinformatics analysis on GeDTC gene was carried out by using ExPASY, ClustalW, MEGA, etc. Positive transgenic plants and potato minituber were obtained by virtue of the potato genetic transformation system. Agronomic characters, such as size, weight, organic acid content, and starch content, of potato minituber were tested and analyzed and GeDTC gene function was preliminarily investigated. The results showed that the open reading frame of GeDTC gene was 981 bp in length and 326 amino acid residues were encoded, with a relative molecular weight of 35.01 kDa. It was predicted that the theoretical isoelectric point of GeDTC protein was 9.83, the instability coefficient was 27.88, and the average index of hydrophilicity was 0.104, which was indicative of a stable hydrophilic protein. GeDTC protein had a transmembrane structure and no signal peptide and was located in the inner membrane of mitochondria. The phylogenetic tree showed that GeDTC was highly homologous with DTC proteins of other plant species, among which GeDTC had the highest homology with DcDTC(XP_020675804.1) in Dendrobium candidum, reaching 85.89%. GeDTC overexpression vector pCambia1300-35Spro-GeDTC was constructed by double digests, and transgenic potato plants were obtained by Agrobacterium-mediated gene transformation. Compared with the wild-type plants, transgenic potato minituber harvested by transplanting had smaller size, lighter weight, lower organic acid content, and no significant difference in starch content. It is preliminarily induced that GeDTC is the efflux channel of tricarboxylate and related to the tuber development, which lays a foundation for further elucidating the molecular mechanism of G. elata tuber development.
		                        		
		                        		
		                        		
		                        			Gastrodia/genetics*
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		                        			Phylogeny
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		                        			Amino Acids
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		                        			Cloning, Molecular
		                        			
		                        		
		                        	
            
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