1.Prospect and Progress of the Research on the Technology of Metabolism in Acupuncture and Moxibustion
Qin CHEN ; Qiaofeng WU ; Jianqiao FANG
Journal of Zhejiang Chinese Medical University 2016;40(3):223-226
Objective] Metabolomics is a genomics research methods, has good prospects in the study of mechanisms of acupuncture. In this paper, the progress of the research on the mechanism of the action of metabolism in the mechanism of acupuncture and moxibustion were reviewed. [Methods] Retrieve acupuncture mechanism studies,which use metabolic technology in nearly 15 years, and to summarize. [Results]Nearly 15 years metabolomics research techniques and tools developed rapidly. But the acupuncture metabomoic studies were still based on the research of animal experiments mainly, relative clinical research less. [Conclusion] With the deepening development of metabolomics studies, the metabolic characteristics of various diseases will gradually clear. Acupuncture clinical mechanisms research combined with metabolomics technology will have broader prospects. But the research strategy of metabololic technology on acupuncture clinical efficacy is yet to be further explored and improved.
2.Biliary manometry in patients with common bile duct stones and juxtapapilary duodenal diverticulum
Fang HAO ; Mingfang QIN ; Yu WU
Chinese Journal of Digestive Endoscopy 2015;32(7):457-461
Objective To explore the impact of juxtapapilary duodenal diverticulum on the formation of choledocholithiasis and biliary manometry after surgery.Methods The clinical data of 97 patients with common bile duct stones who successfully underwent ERCP and biliary manometry treatment from May 2011 to March 2014 were retrospectively analyzed.Ninty-seven common bile duct stone patients were divided to two groups,that is,52 cases of juxta-papilary duodenal diverticulum (experimental group) and 45 cases without juxta-papilary duodenal diverticulum (control group).The biliary manometric results before and after EST of both groups were compared.Results The basal pressure of sphincter Oddi of the experimental group before EST [(84.8 ± 26.1) mmHg] was higher than that of the control group before EST [(75.5 ± 14.6) mmHg] (P < 0.05,1 mmHg =0.133 kPa);and that after EST was also higher than that of the control group after the operation[(19.8 ± 14.2)mmHg VS (15.9 ± 9.8)mmHg,P < 0.05].The biliary pressure of common bile duct stone patients with JPD was higher than those without JPD before and after EST but with no significant difference(P > 0.05).The biliary manometry was performed in 81 patients after one month and it was found that the basal pressure of sphincter Oddi of experimental group was still higher than that of the control group [(34.8 ± 17.1) mmHg VS (29.6 ± 15.3)mmHg,P < 0.05].The biliary pressure of common bile of the experimental group was higher than that of the control group [(19.9 ± 11.2) mmHg VS (17.1 ± 13.1)mmHg,P <0.05].Conclusion The existence of juxtapapilary duodenal diverticulum increases the pressure of common bile duct and Oddi sphincter and leads to CBD stones.
3.Study on Multi-index Quality Evaluation of Atracylodis Macrocephalae Rhizoma Based on PCA Model
Fang LI ; Hongyan WU ; Guotai WU ; Qin LI
Chinese Journal of Information on Traditional Chinese Medicine 2016;23(11):81-85
Objective To establish a multi-index quality evaluation method of Atracylodis Macrocephalae Rhizoma based on principal component analysis (PCA) model.Methods The contents of atractylenolideⅠ and atractylenolideⅢ were determined by HPLC; atractylodes macrocephalaon polysaccharide was determined by phenol-sulfuric acid method; ethanol soluble extractum was determined by hot soak in general rule of Chinese Pharmacopoeia (2015 Edition); PCA was used to establish a multi-index quality evaluation model. Results AtractylenolideⅠ and atractylenolideⅢ showed good linear correlations (r=0.999 9), and the average recoveries were 100.9% and 102.1%, respectively. PCA showed that the Atracylodis Macrocephalae Rhizoma from Panan County, Yuqian Town, Jinyun County, Xinchang County of Zhejiang Province had the best quality, from Bozhou City and Fuyang City of Anhui Province had good quality.Conclusion The established multi-index quality evaluation method based on PCA model can be applied to objectively evaluate the quality of Atracylodis Macrocephalae Rhizoma from different producing areas.
4.Clinical significance of serum calcium-binding protein A9(S100A9)levels in patients with rheumatoid ar- thritis
Wenhua XUAN ; Qin WU ; Yumeng SHI ; Fang WANG ; Wenfeng TAN
The Journal of Practical Medicine 2017;33(16):2658-2661
Objective To investigate the serum level of calcium-binding protein A9(S100A9)in patients with rheumatoid arthritis and explore its potential clinical significance. Methods The serum level of S100A9 was measured by ELISA in 79 rheumatoid arthritis (RA) patients and 20 healthy controls (HC). The correlation of S100A9 level and relevant clinical parameters were analyzed. Results The serum level of S100A9 was significantly increase in RA patients than those in HC. The serum level of S100A9 was higher in high disease activity than that in moderate and low disease activity in the RA group. There was a positive correlation in serum S100A9 level and DAS28 score,Rheumatoid factor,tender joint count and swollen joint count. Conclusion The serum level of S100A9 increases in RA patients and correlates with RA activity.
5.Ag85A DNA vaccination boosting enhances BCG primed-mice anti-tuberculosis T cell responses
Han KANG ; Xiaoyong FAN ; Qin YUAN ; Fuming WU ; Fang SHEN
Chinese Journal of Microbiology and Immunology 2013;(1):66-72
Objective To construct DNA vaccine expressing Mycobacterium tuberculosis(Mtb) immunodominant antigen Ag85A and analyze its anti-tuberculosis T cell responses in BCG primed-mice after DNA vaccination boosting.Methods The coding gene of Ag85A mature fragment was amplified by PCR with H37Rv genomic DNA as template,and then cloned into the eukaryotic expression vector pVAX1 to construct Ag85A DNA vaccine.After purification,Ag85A DNA vaccine was injected intramuscularly twice in BCG primed-mice with BCG vaccination and DNA vaccination alone as control.Eight weeks post-vaccination,spleen lymphocytes were separated and were then used to analyze Mtb antigen specific effector T cell response and polyfuntional IFN-γ/TNF-α/IL-2 secreting CD4+ T cell frequencies and intensities,and CD8+T cell responses by IFN-γ ELISPOT assay and intracellular staining,respectively.Results Compared to BCG vaccinated-and DNA vaccinated-mice,Ag85A DNA boosting not only enhanced significantly BCG primed-mice IFN-γ+TNF-α+IL-2+,IFN-γ+ IL-2+,TNF-α+IL-2+ and IL-2+ CD4+ T cell frequencies and IL-2 secretion,but also improved significantly IFN-γ-secreting and IL-2-secreting CD8+ T cell frequencies.Condusion Ag85A DNA vaccine was constructed successfully and was demonstrated to enhance significantly BCG primed-mice Mtb antigen specific CD4+ and CD8+ T cell responses when boosting,which is beneficial to improve BCG immunogenicity and its waning immune protection against Mtb.
6.Role of serum folate, polymorphisms related reduced folate carrier gene and methionine synthase reductase gene in cervical cancer
Fang CHEN ; Jintao WANG ; Ling DING ; Qin ZHOU ; Yuanyuan WU
Cancer Research and Clinic 2013;25(7):437-440
Objective To evaluate the possible association among serum folate,polymorphisms related reduced folate carrier gene (RFC-1) AS0G,methionine synthase reductase gene (MTRR) A66G,and cervical cancer,and to provide clues for the etiology of cervical cancer.Methods Based on a hospital-based case-control study,107 cases diagnosed as cervical cancer pathematologically and 107 controls with hysteromyoma,were selected by frequency,matched with age and habitation.Serum folate concentration was detected by RIA and polymorphism RFC-1 A80G and MTRR A66G was examed by RFLP-PCR.Results Serum folate concentration in patient group [(1.86±0.60) ng/rml] was significantly lower than that in control group [(2.30 ± 1.14) ng/ml],and risk of cervical cancer increased with the decreased serum folate levels (x2trend =12.57,P =0.001).Risks to catch cervical cancer for women with RFC-1 80 GG were 2.42 times (95 % CI 1.01-5.81) as much as for those with RFC-1 80 AA,and 1.65 times (95 % CI 0.77-3.53) for those with RFC-1 80 AA and RFC-1 80 AG,risks to catch cervical cancer for women with MTRR 66 GG were 1.35 times (95 % CI 0.40-4.56) as much as for those with MTRR 66 AA and 1.26 times (95 % CI 0.38-4.16) for those with RFC-1 80 AA and RFC-1 80 AG.Conclusion Serum folate deficiency to a certain degree can increase the risk of cervical cancer.RFC-1 A80G mutation may be a risk factor for cervical cancer and homozygous (GG) gene may increase the susceptibility of cervical cancer.MTRR A66G gene mutation may have nothing to do with cervical cancer.
7.Protective Effect and Mechanism of Human Lipoxin A4 on N2a Cell Injury Induced by β-Amyloid Protein 25-35
Qiang WU ; Le WU ; Zhipeng XU ; Min CUI ; Jie PU ; Fang CHEN ; Qin LIU
China Pharmacist 2017;20(8):1340-1344
Objective: To investigate the protective effect of human lipoxin A4 (LXA4) on N2a cell damage induced by β-amyloid protein 25-35 (Aβ25-35) and the underlying mechanism. Methods: Aβ25-35 was used to treat N2a cells to establish Alzheimer's disease (AD) cell injury model. Meanwhile, LXA4 was added to the experimental group at different concentrations (50, 100 and 200 nmol·L-1 ). MTT assay was used to detect the activity of N2a cells. The apoptosis was detected by Hoechst 33258-PI staining, the expression of P62 and TRAF6 mRNA was detected by RT-PCR, and the expression of P62 and TRAF6 protein was detected by Western blot. Results: Compared with that of the model group, the cell survival rate of LXA4 protective group (50,100 and 200 nmol·L-1 ) increased (P <0. 01) and the apoptosis of N2a cells induced by Aβ25-35 was reduced by LXA4 (100 and 200 nmol·L-1 ) . Compared with that of the model group, the expression of P62-mRNA and protein-P62 of N2a cells treated with Aβ25-35 increased (P <0. 05 or P <0. 01) and the expression of TRAF6-mRNA and protein-TRAF6 of N2a cells treated with Aβ25-35 were reduced (P <0. 05 or P <0. 01). Conclusion: LXA4 has protective effect on N2a cell damage induced by Aβ25-35 , and its mechanism may be related to the up-regulation of P62 gene and down-regulation of TRAF6 gene.
8.Puerarin extraction and identification of solution wine to protect liver function in mice
Jing WANG ; Hongmin LI ; Fang AI ; Xiong CAO ; Aijuan WU ; Qin HAN
Journal of Regional Anatomy and Operative Surgery 2015;(4):358-361
Objective Pueraria extract puerarin,HPLC assay puerarin extract and compare different doses of correlation with the hang-over effect of puerarin evaluate different doses of puerarin liver hangover effect. Methods Extracted under optimal conditions obtained in the previous experiment puerarin spare,HPLC method for qualitative and quantitative detection of alcohol extract of kudzu root ( PRE) ,the male Kunming mice were randomly divided into control group,positive control group and puerarin group,each group of 10. Give mice fed pueraria extract,30 min after administration of liquor,drunk mice sobering observation time and the determination of mouse liver ADH,GOT,GPT con-tent in order to investigate the effect of puerarin on drunken mice. Results HPLC fraction was measured at 8 times the volume of 70% etha-nol,60 ℃ constant temperature water bath shaker at 30 min for optimal extraction conditions puerarin extraction. Compared with the positive control groups:low,medium and high doses of alcohol extract of pueraria can significantly shorten the time to sober up drunken mice,the dose of PRE could effectively inhibit the absorption of alcohol,reduce liver tissue ADH,GOT,GPT,the effects of high doses of PRE absorption of alcohol was small. Conclusion HPLC method capable of puerarin extract the qualitative and quantitative determination of puerarin on liver injury caused by acute alcoholism a protective regulatory role,and the hangover effect of puerarin dose showed a good positive correlation.
9.The study on the construction of Pluronic P85 coated poly(butylcyanoacrylate) nanoparticles and its brain target effect on a rat model of mesial temporal lobe epilepsy
Ziyan FANG ; Caifeng GUO ; Fengchun WU ; Jiaming QIN ; Yuping NING ; Liemin ZHOU
Chinese Journal of Nervous and Mental Diseases 2017;43(6):356-361
Objective In order to evaluate that whether Pluronic P85 coated poly(butylcyanoacrylate) nanoparti-cles was able to deliver antiepileptic drug phenytoin into the brain va bypassing mesial temporal lobe epilepsy (MTLE)-induced Pgp in a rat model of MTLE. Methods The rat model of MTLE, induced by li-pilocarpine, was divided in-to two groups (6 for nanoparticle drug group and 7 for PHT drug group). Immunohistochemistry assay was performed to detect Pgp expression at the hippocampus. Nanoparticles were prepared by interfacial polymerization method. Dialysate samples of brain were collected at 30, 60, 120, 180, 240 and 300 min after drug administration by microdialysis tech-nology. Samples were analyzed by high performance liquid chromatography (HPLC). Results The area under the curve (AUC) ratio of brain/plasma in Nanoparticle drug group was 0.370.10 which was significantly higher compared with 0.190.06 in conventional PHT drug group (P<0.05). The Pgp immunopositive area, as assessed by analysis of labeled surface area, was higher in the DG, CA3 and CA1 sector in the hippocampus of MTLE rats when compared to the normal rats. Conclusions Pluronic P85 coated PBCA nanoparticles can significantly deliver PHT into brain via bypassing MTLE-induced Pgp in a rat model of MTLE.
10.Effects of erythropoietin in high glucose induced proliferation and apoptosis of human kidney proximal tubular epithelial cells and the possible mechanism
Yanxia CHEN ; Xianfeng WU ; Xiangdong FANG ; Xiaohua QIN ; Chong HUANG ; Weiping TU
Tianjin Medical Journal 2015;(1):25-29
Objective To study the effects of erythropoietin (rhEPO) in high glucose induced proliferation and apopto?sis of human kidney proximal tubular epithelial (HK-2) cells, and the possible mechanism thereof. Methods HK-2 cells cultured in vitro were divided into several groups randomly:blank control group, high glucose group, mannitol group, rhEPO control group, different concentrations of rhEPO treatment groups (5, 10, 20 U/mL) and Rho kinase group. The reverse tran?scription polymerase chain reaction (RT-PCR) was used to evaluate the mRNA levels of RhoA and ROCK after 24 hours. Tetrazolium salt method (MTT) was used to determine the cell proliferation. Cell apoptosis was detected by flow cytometry. Results Compared with blank control group the expression levels of RhoA and ROCK1 mRNA were significantly in?creased in high glucose group (P < 0.05). RhoA, ROCK1 mRNA expressions significantly decreased in rhEPO group than those of high glucose group (P<0.05). There was a positive correlation between the expression levels of RhoA mRNA and ROCK1 mRNA in high glucose group and rhEPO group. MTT method showed that rhEPO significantly promoted the prolifer?ation of HK-2 cells (P<0.05). Flow cytometry analysis showed that high glucose induced apoptosis in HK-2 cells, which was significantly inhibited in rhEPO group and Rho kinase group as compared to that of high glucose group in a concentra?tion dependent manner (P<0.05). Conclusion rhEPO can promote HK-2 cell proliferation and inhibit apoptosis, which may be related to RhoA/ROCK signaling pathway.