1.Effects of electroacupuncture on motor function and related molecular mechanisms in mice with Parkinson's disease
Ling QI ; Yanan LI ; Yao WANG ; Xiaolei ZHANG ; Mengni HU ; Hanzhang LI ; Die XIAO ; Zhen RONG ; Jun MA
Journal of Beijing University of Traditional Chinese Medicine 2024;47(5):721-728
Objective To explore the effects of electroacupuncture on motor function in Parkinson's disease(PD)model mice and NLRP3 inflammasome-related proteins in the midbrain substantia nigra(SN).Methods C57BL/6 mice were assigned to three groups according to the random number table method:control group,model group,and electroacupuncture(EA)group,12 mice per group.The PD model was reproduced by intragastric administration of rotenone solution 10 mg/(kg·d).EA group was administered at the three selected points,"Fengfu"(GV16),"Taichong"(LR3),and"Zusanli"(ST36),with a treatment cycle of 2 weeks.The control and model groups took the same time synchronous fixation operation for the control variable.Behavioral scores and open field tests were used to detect the exercise ability of mice in each group.Tyrosine hydroxylase(TH)and α-synuclein(α-syn)in the midbrain SN of mice in all groups were measured with an immunohistochemistry test.NLRP3 and cysteinyl aspartate specific proteinase-1(Caspase-1)protein expression levels in the midbrain SN of mice in the three groups were measured using Western blotting,and interleukin-1β(IL-1β)content was determined with an enzyme-linked immunosorbent assay.Results Compared to the control group,the behavioral scores of the mice in the model group were higher(P<0.01).Compared to the model group,the behavioral scores of the mice in the EA group were lower(P<0.01).Compared to the control group,the time ratio of the relative rest state of the mice in the model group(<100 mm/s)increased significantly(P<0.01),while the time ratio of the slow motion(100~200 mm/s)and time ratio of the fast motion(>200 mm/s)state decreased significantly(P<0.01).Compared to the model group,the time ratio spent in the relative rest state of mice in the EA group decreased significantly(P<0.01),while the time ratio of the slow motion state and time ratio of the fast motion state and movement rate increased significantly(P<0.01).Compared to the control group,the TH expression level decreased in the SN in the model group(P<0.01),while α-syn increased(P<0.01).Compared to the model group,the TH expression level in the EA group increased(P<0.05),while α-syn decreased(P<0.05).Compared to the control group,the protein expressions of NLRP3 and Caspase-1 in the SN of the model group increased(P<0.01);compared to the model group,the expressions of NLRP3 and Caspase-1 in the SN of the midbrain of mice decreased after EA treatment(P<0.01).Compared to the control group,IL-1β in the SN of the mouse midbrain increased in the model group(P<0.01).Compared to the model group,IL-1β decreased in the EA group(P<0.05).Conclusion This experiment shows that stimulation of EA in"Fengfu","Taichong",and"Zusanli"can effectively reduce abnormal aggregation of the PD marker α-syn,increase TH expression,and enhance the motor dysfunction of PD model mice.The molecular mechanism is related to the regulation of the expression of NLRP3,Caspase-1,and IL-1β of inflammasome-related pathways.
2.Effect and mechanism of cinnamon oil on rat prostatic hyperplasia
Wei HE ; Die SONG ; Hong-Ping WANG ; Song-Tao MA
The Chinese Journal of Clinical Pharmacology 2024;40(9):1292-1295
Objective To research the effect and the mechanism of cinnamon oil on experimental rat prostatic hyperplasia.Methods SD rats were randomly divided into normal group,model group,control group and cinnamon oil-H,-M,-L groups,with 10 rats in each group.The prostatic hyperplasia model was induced by injected subcutaneously with 5 mg·kg-1·d-1 testosterone propionate for 30 d.Normal group and model group were given 1 mL·kg-1 0.9%NaCl;control group were given 1 mg·kg-1 finasteride;cinnamon oil-H,-M,-L groups were given 1,0.5,0.25 mg·kg-1 cinnamon oil by intragastric administration,respectively,once a day for 30 d.The wet weight and index of prostate gland in rats were compared to explore the therapeutic effect of cinnamon oil,and the inhibitory effect of cinnamon oil on 5α-reductase and NADPH oxidase was studied to explore its mechanism.Results The prostate wet weight in normal group,model group,control group and cinnamon oil-H,-M,-L groups were(0.51±0.09),(1.02±0.17),(0.65±0.15),(0.64±0.17),(0.79±0.09)and(0.81±0.15)mg;the prostate index were(2.25±0.33),(4.70±0.48),(3.03±0.82),(3.02±0.60),(3.53±0.46)and(3.68±0.66)mg·kg-1;the fluorescence intensity of NADPH oxidase were(1.26±0.18),(5.21±0.80),(2.21±0.56),(2.51±0.56),(4.13±0.64)and(4.90±0.88)RLU,respectively.Compared with model group,the fluorescence intensity of NADPH oxidase in normal group,control group and cinnamon oil-H,-M groups all had significantly differences(all P<0.01).The average inhibition rates of 5α-reductase in positive control group(20 μmol·L-1),cinnamon oil group(10,20 μmol·L-1)were 49.36%,45.62%and 63.45%,respectively.The average inhibition rate of 5α-reductase in cinnamon oil group(20 μmol·L-1)was higher than that in positive control group(20 μmol·L-1),the difference was statistically significant(P<0.05).Conclusion Cinnamon oil may inhibit prostate hyperplasia by inhibiting 5α-reductase and NADPH oxidase.
3.Blood transfusion quality supervision in a district of Shanghai
Hui ZHANG ; Caili WEN ; Die SUN ; Qun MIAO ; Lijuan MA ; Lin GUAN ; Yiming YANG ; Xiaowen CHENG
Chinese Journal of Blood Transfusion 2022;35(5):575-579
【Objective】 To supervise the clinical blood use of 19 hospitals, covering a district of Shanghai, during two years, and discover the problems in the process of blood transfusion, so as to put forward suggestions for corrective methods in grades and promote continuous improvement of clinical transfusion management. 【Methods】 A total of 19 hospitals were supervised in terms of hardware facilities, management level, professional and technical level, and blood typing test on the site, according to the Administrative Blood Management Measures for Medical Institutions, Technical Specification for Clinical Transfusion and Shanghai Medical Quality Supervision Score Statistical Table.All data were analyzed. 【Results】 These hospitals can properly perform clinical blood transfusion, but there were obvious differences.Tertiary hospitals were relatively better, yet need to strengthen the management of medical documents.Secondary hospitals remained to be improved, mainly in insufficient construction of Blood Transfusion Department (blood bank), the lack of management and maintenance of key equipment and the lack of standardization in medical documents writing.However, flaws in the supervision were general in private hospitals (most of which were affiliated hospitals), so the management of clinical blood use should be further strengthened. 【Conclusion】 For secondary hospitals or above, routinized writing of medical documents and promoted construction of Blood Transfusion Department (blood bank) should be strengthened.For private hospitals, especially affiliated hospitals, the management of clinical blood use should be further improved, including the examination rules corresponding to the blood use process and strict access and exit mechanism, so as to improve the overall management level of clinical blood use and ensure the safety of clinical blood use.
4.Effects of the expression of serum SIRT1 on therapeutic efficacy of sodium valproate in the treatment of epilepsy patients
Yanping YANG ; Die HE ; Zhen ZHOU ; Qingfan ZENG ; Hongying PENG ; Lixin ZHAO ; Dan MA
China Pharmacy 2022;33(23):2886-2890
OBJECTIVE To explore the effects of the expression of serum Sirtuin-1 (SIRT1) on therapeutic efficacy of sodium valproate (VPA) in the treatment of epilepsy patients. METHODS Fifty-four epileptic patients were collected from the Affiliated Baiyun Hospital of Guizhou Medical University from Mar. to Oct. 2021 as the research objects, and fifty healthy people were also collected during corresponding period as baseline reference samples. The patients whose relative mRNA expression of SIRT1 was lower than the baseline were selected as SIRT1 low-expression treatment group, and the patients whose that expression was higher than the baseline as SIRT1 high-expression treatment group.All patients were treated with low dose (12 mg/kg or about 600 mg/day) of VPA for 3 months, and the clinical efficacy was evaluated. The dosage of VPA in patients with ineffective gzwkj2021-468) epilepsy control should be increased to 15 mg/kg or about 800 mg/day for another 3 months as SIRT1 high-expression intensive treatment group and SIRT1 low-expression intensive treatment group. Clinical efficacies were evaluated. The blood concentration and liver function indexes of the patients were detected after 3 months of treatment and 3 months of intensive treatment. RESULTS Before treatment, among 54 epileptic patients, 31 epileptic patients had low expression of SIRT1, and 23 had high expression of SIRT1. After 3 months of low-dose VPA treatment, within effective blood concentration of VPA, effective control rate of patients in SIRT1 high-expression treatment group was significantly lower than SIRT1 low-expression treatment group (P<0.05). After 3 months of intensive treatment, the effective control rate of patients SIRT1 high-expression intensive treatment group was significantly higher than SIRT1 high expression treatment group (P<0.05). No abnormality was found in liver function indexes during VPA treatment. CONCLUSIONS Epilepsy in patients with high expression of serum SIRT1 may be more difficult to control when VPA is within the effective blood concentration range; when the VPA dose is effectively increased, the effective control rate of epilepsy can be improved.
5.Effect of therapies for kidney-tonifying and blood-activating in treatment of anovulatory infertility in eugenics.
Kun MA ; Cai-Die TIAN ; Yan-Xia CHEN ; Min LI ; Lin-Juan GONG
China Journal of Chinese Materia Medica 2021;46(11):2634-2638
In the context of the new era, paying attention to maternal and child health and advocating prenatal and postnatal care can effectively improve the quality of the birth population. Traditional Chinese medicine has a long history of prenatal and postnatal healthcare with rich content, which is the theoretical basis of modern related services. With the social development and the improvement of people's awareness of prenatal and postnatal healthcare, people have gradually shifted the focus of prenatal and postnatal healthcare to the peri-pregnancy stage at present, namely that couples of childbearing age are guided to prepare for pregnancy under the premise of solving their basic diseases. Infertility is a common and refractory disease for women of childbearing age. Ovulation disorder is one of its common pathological mechanisms. Traditional Chinese medicine believes that kidney deficiency is the main cause and pa-thogenesis of anovulation infertility and blood stasis is an important factor throughout the disease course. In clinical practice, therapies for invigorating kidney and activating blood are safe and reliable to treat anovulatory infertility mainly by adjusting the hypothalamus-pituitary-ovarian axis, improving ovarian function, uterine environment and gamete quality and increasing endometrial volume. Under the guidance of the thought of prenatal and postnatal healthcare, the authors tried to explore the effect of therapies for kidney-tonifying and blood-activating in the treatment of anovulatory infertility in eugenics, with the purpose of providing ideas and basis for subsequent relevant clinical studies and contributing to prenatal and postnatal healthcare services.
Anovulation
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Child
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Eugenics
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Female
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Humans
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Infertility, Female/drug therapy*
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Kidney
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Medicine, Chinese Traditional
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Ovulation
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Pregnancy
6.Study on mechanism of Bushen Culuan Formula in treatment of polycystic ovary syndrome based on network pharmacology and molecular docking.
Kun MA ; Lin-Juan GONG ; Yan-Xia CHEN ; Cai-Die TIAN
China Journal of Chinese Materia Medica 2021;46(11):2650-2659
This study used network pharmacology and molecular docking to study the mechanism of Bushen Culuan Formula in the treatment of infertility caused by polycystic ovary syndrome(PCOS). The active ingredients and potential drug targets of Bushen Cu-luan Decoction were obtained by searching the Traditional Chinese Medicine System Pharmacology(TCMSP) database, and the targets of PCOS by searching GeneCards. After the drug targets and disease targets were corrected by Uniprot, the intersection genes were obtained. STRING database and Cytoscape 3.7.2 were used for protein-protein interaction(PPI) analysis of the intersection genes. The ClueGO plug-in of Cytoscape 3.7.2 was employed to perform gene ontology(GO) enrichment and KEGG pathway enrichment for the intersection genes. Finally, molecular docking of the key active ingredients with the targets of Bushen Culuan Formula was performed using AutoDockVina and MGLtools. A total of 136 active ingredients and 314 drug targets of the decoction were obtained from TCMSP, and 136 disease targets from GeneCards. Finally, 49 drug-disease intersection genes were obtained. GO enrichment found that the genes were mainly involved in the regulation of muscle cell apoptosis, positive regulation of small molecule metabolism, core promoter binding, RNA polymerase Ⅱ regulation of pri-miRNA transcription, negative regulation of transmembrane transport and other biological functions. The enriched KEGG pathways mainly included MAPK, PI3 K-Akt, p53, and HIF-1 signaling pathways. The results of molecular docking showed that quercetin and PTGS2 can bind stably and interact through amino acid residues THR206, TRP387, ASN382, etc. This study preliminarily reveals the multi-component, multi-target, and multi-pathway mechanism of Bushen Culuan Formula in the treatment of PCOS-related infertility, which provides a basis for further research.
Drugs, Chinese Herbal/pharmacology*
;
Female
;
Gene Ontology
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Humans
;
Medicine, Chinese Traditional
;
Molecular Docking Simulation
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Polycystic Ovary Syndrome/genetics*
;
Signal Transduction
7.Effect of Dingkundan on Sex Hormone and Follicle Count in Mice of Tripterygium Wilfordii Polyglycosides Induced Diminished Ovarian Reserve
Yan-xia CHEN ; Yuan YUAN ; Kun MA ; Xiao-di FAN ; Kai-li WANG ; Cai-die TIAN ; Min LI
Chinese Journal of Experimental Traditional Medical Formulae 2020;26(14):78-84
Objective:To establish a mouse model of diminished ovarian reserve (DOR) induced by tripterygium wilfordii polyglycosides (TWP), and to explore the therapeutic effect of Dingkundan (DKD) on DOR, so as to provide scientific basis for its clinical application. Method:The 60 female Blab/c mice with regular estrous cycle were randomly divided into blank group, model group, low,medium and high-dose DKD group, DKD group and estradiol valerate group, with 10 mice in each group. Except the blank group, the other groups were given 40 mg·kg-1 TWP suspension. Meantime,low,medium and high-dose DKD group were given 1.64,3.28,6.56 g·kg-1 DKD suspension respectively, and estradiol valerate group was given 0.15 mg·kg-1 estradiol valerate suspension by gastric lavage once a day for 30 days. The general condition, body weight, estrous cycle and gonad index of mice were observed, serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) were determined by radioimmunoassay, ovarian morphology and follicle count were observed by hematoxylin-eosin staining (HE) staining. Result:Compared with the blank group, most of the mice in model group had disordered estrous cycle, uterine and ovarian indexes decreased (
8.Establishment of premature ovarian insufficiency kidney deficiency and blood stasis pattern mouse model with Tripterygium wilfordii polyglycoside for Bushen Culuan Decoction therapy.
Yuan YUAN ; Yan-Xia CHEN ; Kun MA ; Bo-Chao YUAN ; Kai-Li WANG ; Cai-Die TIAN
China Journal of Chinese Materia Medica 2019;44(9):1895-1903
To establish a mouse model of premature ovarian insufficiency( POI) with kidney deficiency and blood stasis pattern by Tripterygium wilfordii polyglycoside( TWP) gavage,and to evaluate the ovarian function and fertility of the model,in order to find Bushen Culuan Decoction therapeutic mechanism. 60 SPF level Blab/c female mice with normal estrous cycle were randomly divided into 6 groups of 10 each: blank group 1( BG1),blank group 2( BG2),blank fertility group( BFG),model group( MG),model recovery group( MRG) and model fertility group( MFG). The mice in three model groups were treated by gastric gavage with TWP suspension 40 mg·kg-1 twice a day for 14 days,while the mice in three blank groups were treated by gastric gavage with same volume normal saline for 14 days. The mice in BG1 and MG were sacrificed and dissected on day 15. The mice in BG2,BFG,MRG and MFG were returned normal feeding from day 15 and were sacrificed and dissected on day 29. The mice in BFG and MFG were cohabited with male mice with a ratio of 2 ∶1( female ∶male) from day 15. The general situation and estrous cycles of all mice were observed every day. Serum sex hormone levels,ovarian index,uterine index,ovarian morphology,follicle count,ovarian VEGF and ES index were observed within the mice in BG1,BG2,MG and MRG. Pregnancy rate,litter size,survival number of newborn mice and male-female proportion were reported within the mice in BFG and MFG. In model establishing stage,the body weight of mice significantly decreased( P <0. 05) in MG and MFG. Compared with BG1,the mice in model group had irregular estrous cycle,decreased ovarian and uterine indexes,less primordial and developing follicles,more atretic follicles,increased VEGF expression and decreased ES expression( P <0. 05). Compared with blank group 2,the mice in model recovery group had irregular estrous cycle,increased FSH level,decreased ovarian indexes,less primordial and developing follicles,more atretic follicles,increased VEGF expression( P<0. 05). Compared with blank fertility group,the mice in model fertility group had smaller litter size and newborn mice survival count( P<0. 05). Gastric gavage with TWP 40 mg·kg-1 twice a day for 14 days is a feasible way to establish a POI kidney deficiency and blood stasis pattern mouse model. The mice ovarian functions didn't recovery on day 14 after stopping TWP intervening,which could suggest the effectiveness of subsequent therapeutic intervention.
Animals
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Disease Models, Animal
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Drugs, Chinese Herbal
;
pharmacology
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Female
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Mice
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Mice, Inbred BALB C
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Pregnancy
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Primary Ovarian Insufficiency
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chemically induced
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drug therapy
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Random Allocation
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Tripterygium
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adverse effects
9.Cloning and functional identification of a new NADPH-cytochrome P450 reductase in Andrographis paniculata.
Meng-Die QI ; Jian WANG ; Xiao-Jing MA ; Quan ZHANG ; Fang-Fang WANG ; Ying KANG ; Hui-Xin LIN ; Yong LIU
China Journal of Chinese Materia Medica 2018;43(2):309-315
Andrographolide is a main active ingredient in traditional Chinese medicine Andrographis paniculata,with a variety of pharmacological activity,widely used in clinical practice. However its biosynthetic pathway has not been resolved. Cytochrome P450 reductase provides electrons for CYP450 and plays an important role in the CYP450 catalytic process. In this study,the coding sequence of A. paniculata CPR was screened and cloned by homologous alignment,named ApCPR4. The ApCPR4 protein was obtained by prokaryotic expression. After isolation and purification,the enzyme activity was identified . The results showed that ApCPR4 could reduce the cytochrome c and ferricyanide in NADPH-dependent manner. In order to verify its function,ApCPR4 and CYP76AH1 were co-transformed into yeast engineering bacteria. The results showed that ApCPR4 could help CYP76AH1 catalyze the formation of rustols in yeast. Real-time quantitative PCR results showed that the expression of ApCPR4 increased gradually in leaves treated with methyl jasmonate (MeJA). The expression pattern was consistent with the trend of induction and accumulation of andrographolide by MeJA,suggesting that ApCPR4 was associated with biosynthesis of andrographolide.
Acetates
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Andrographis
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enzymology
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genetics
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Biosynthetic Pathways
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Cloning, Molecular
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Cyclopentanes
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Diterpenes
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metabolism
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NADPH-Ferrihemoprotein Reductase
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genetics
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Oxylipins
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Plant Leaves
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enzymology
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Plant Proteins
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genetics
10.Overexpression and isolation of CYP76AH1 in Salvia miltiorrhiza hairy roots.
Meng-Die QI ; Jian WANG ; Xiao-Jing MA ; Quan ZHANG ; Fang-Fang WANG ; Ying KANG ; Yong LIU ; Hui-Xin LIN
China Journal of Chinese Materia Medica 2017;42(20):3901-3905
Protein complexes are involved in the synthesis of multiple secondary metabolites in plants, and their separation is essential to elucidate plant secondary metabolism and improve in vitro catalytic efficiency. In this study, the transgenic hairy roots of CYP76AH1, a key enzyme of tanshinone synthesis pathway, was constructed and the transgenic hairy roots of Danshen overexpressing CYP76AH1 protein were screened by Western blotting and used as a tissue culture material for the subsequent extraction of protein complex in tanshinone synthesis pathway. By optimizing the type and concentration of the detergent in the protein extraction buffer, the buffer containing 0.5% Triton X-100 was selected as the best extraction buffer, and a relatively large amount of soluble CYP76AH1 protein was isolated. This study lays the foundation for the further separation and purification of protein complexes interacting with CYP76AH1, and provides the idea for deep analysis of tanshinone metabolic pathway.

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