1.The mechanism of action of valsartan studied by HPLC-TOF/MS.
Wenqing YANG ; Yunlun LI ; Haiqiang JIANG
Acta Pharmaceutica Sinica 2015;50(7):875-81
High performance liquid chromatography-time-off-flight mass spectrometer (HPLC-TOFMS) technology coupled with partial least squares discriminant analysis (PLS-DA) processed by SIMCA-P software was applied to investigate serum endogenous metabolites alternations of valsartan in spontaneous hypertension rats (SHR). And MetPA platform was used to connect identified potential biomarkers in corresponding metabolic pathways to find possible therapeutic mechanism of valsartan. Valsartan significantly declined the blood pressure of SHRs (P < 0.05) at fourth week. The metabolic profiling significantly changed and four metabolites involved in G protein-coupled pathway were identified. Metabolomics is able to detect holistic and microcosmic alternations in organism, so as to elucidate therapeutic mechanism of drugs.
2.Correlation of Cav-1 mRNA, S100A4 mRNA and CD31 expression with prostate cancer tumor metastasis and patient survival rate
Daina ZHUANG ; Haiqiang YAN ; Zuisu YANG ; Guofang DING
Chinese Journal of Urology 2012;33(10):786-790
Objective To explore the expression of Cav-1 mRNA,S100A4 mRNA and CD31 in prostate cancer (PCa) and their correlation with tumor metastasis and patient survival rate. Methods PCa specimens (n =42) and adjacent tissue specimens ( n =12 ) from radical prostatectomy were obtained from January 2004 to May 2006.The mean age of patient was 71.6 ± 7.6 years ( range 58 - 86 years).According to Gleason scores,prostatectomy specimens were stratified into≤6 (n =17),7 (n =12) and ≥8( n =13 ) groups.Patients were classified as clinical stage T1 ( n =16),stage T2 ( n =9 ),stage T3 ( n =11 )and stage T4 (n =6).Patients were divided into PCa with bone metastasis (n =8 ) and PCa without bone metastasis ( n =34).Preoperative PSA levels of the patients were stratified into three groups: < 4 ( n =4)μg/L,4-10 (n=10)μg/Land >10 μg/L (n=28).12 adjacent tissues 1 -2 cm away from tumor or another lobe of prostate were microscopically verified without cancer cells and were tested for comparison.The expression of Cav-1 mRNA and S100A4 mRNA were detected by Situ hybridization in 42 PCa specimens and 12 adjacent tissues and using CD31 for marking vascular endothelial cells,the tumor microvascular density (MVD) was counted.The correlation of Cav-1 mRNA,S100A4 mRNA and CD31 expression was analyzed in combination.with clinical and pathological fcatures including Gleason score,TNM staging,PSA values and bone metastasis. Results The positive expression rate of Cav-1 mRNA in PCa was 35.7% ( 15/42),while it was 0% (0/12) in controls,P <0.05.The positive expression rate of S100A4 mRNA in PCa was 47.6% (20/42),while it was 8.3% (1/12) in controls,P <0.05.The positive expression rate of Cav-1 mRNA in PCa was positively correlated with Gleason score,TNM stage and bone metastasis.The positive expression rate of S100A4 mRNA in PCa was positively correlated with TNM stage and bone metastasis.The average MVD in patients of negative expression of Cav-1 mRNA was (62.8 ± 10.4)/mm2,and the average -MVD in patients of positive expression of Cav-1 mRNA was (83.5 ±6.7 )/mm2,P < 0.05.While the average MVD in patients of negative expression of S100A4 mRNA was (63.3 ± 12.0)/mm2,and the average MVD in patients of positive expression of S100A4 mRNA was (77.9 ± 11.0)/mm2,P < 0.05.The 5-year survival rate in patients with positive Cav-1 mRNA expression was significantly lower than that of with negative expression (46.7% versus 85.2%,P < 0.05 ),while the 5-year survival rate in the patients with positive expression of S100A4 mRNA was significantly lower than that of with negative expression (50.0% versus 90.9%,P < 0.05 ). Conclusions The positive Cav-1 mRNA and S100A4 mRNA expression,increased MVD are positively correlated with PCa progression and bone metastasis. Furthermore,Cav-1 and S100A4 in PCa may promote angiogenesis and cause tumor cells to bone metastases,which can reduce survival rate of patients.
3.Determination of Impurities Including Polymers in Latamoxef Sodium by HPSEC
Xiaoyu LIU ; Shu JIN ; Haiqiang QIU ; Minzhi YANG
China Pharmacist 2015;(2):247-249,270
Objective:To establish a high performance size-exclusion chromatography ( HPSEC) method for the determination of impurities including polymers in latamoxef sodium. Methods:The analysis was performed on a Zenix SEC-150 column(7. 8 mm × 300 mm, 3 μm)with the mobile phase of 0. 005 mol·L-1 phosphate buffer solution [0. 005 mol·L-1 disodium hydrogen phosphate-0. 005 mol·L-1 sodium dihydrogen phosphate (61∶39), pH 7. 0] at a flow rate of 0. 8 ml·min-1. The detection wavelength was set at 254 nm. The column tempretrue was 25℃ and the injection volume was 10μl. Results:The impurities including polymers in latamoxef so-dium were completely separated from latamoxef. The linear range of latamoxef was 0. 98-97. 73 μg·ml-1(r=0. 999 9). The limit of quantitation of latamoxef was 2. 9 ng, and the detection limit was 1. 0 ng. The linear range of the total impurities was 0. 45-2. 8 mg· ml-1(r=0. 999 5). Conclusion: The established method is accurate, rapid and reproducible, and suitable for the determination of impurities including polymers in latamoxef sodium.
4.Intervention effect of Captopril on serum of spontaneous hypertension rats based on metabonomic research
Wenqing YANG ; Haiqiang JIANG ; Yunlun LI ; Yanping ZANG ; Yanjun CHU
Chinese Pharmacological Bulletin 2016;32(7):998-1003
Aims To analyze the intervention effect of Captopril on serum endogenous metabolites alternations in spontaneous hypertension rats ( SHR) and to investi-gate possible therapeutic mechanism .Methods The rapid resolution liquid chromatography/quadrupole-time of flight tandem mass spectrometry ( RRLC-Q-TOF-MS) and technology coupled with partial least squares discriminant analysis ( PLS-DA ) processed by SIMCA-P software were used to distinguish significantly different variables and identify potential biomarkers . Results Compared to the normal group , metabolic profiling changed significantly in model group and Cap-toril group .Totally 4 metabolins and their metabolic pathways were detected , which were closely related to the endothelial function .Conclusion Metabolomics reveals possible therapeutic mechanism of Captopril for protecting endothelial function from overall metabolism of the body .It also shows unique potential in terms of interpretation of the complex mechanisms of drugs .
5.Isolation and purification of an anticancer activity peptide from protein hydrolysate of Mytilus coruscus
Yongfang YANG ; Haiqiang YAN ; Guofang DING ; Fangfang HUANG
Journal of China Pharmaceutical University 2011;42(3):272-275
In this studies, an anticancer peptide was isolated from the enzymatic hydrolysate of Mytilus coruscus.CCK-8 assay was used as guide in activity screening through the isolation process of membrane filtration, DEAEsepharose chromatography, Sephadex G-25 exclusive chromatography and RP-HPLC. An anticancer peptide was obtained and identified as Asp-Leu-Tyr.
6.The teaching model research of medical professionalism
Bo QU ; Haiqiang GUO ; Yang ZHANG ; Shisi LUN ; Baozhi SUN
Chinese Journal of Medical Education Research 2005;0(05):-
In recent years,the health care delivery system changes,the introduction of market mechanisms,as well as health system reform in the traditional medica and so on,have formed a strong spirit of challenge and made a great threat to the nature of medicine and values in the world.Therefore,it is important to teach reform on the physician-patient communication and built a harmonious society.This paper has analyzed the current status and causes of medical education in the spirit of the medical professionalism,and explored the teaching model.
7.The Study on Cross Validation Method for the Split Constituent of the Material fromAstragalus Sweet and Warm Property
Haiqiang JIANG ; Erdong CHEN ; Hongjie CAO ; Yong YANG ; Lili GONG ; Rong RONG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(3):587-590
Whether there was crossing between Astragalus major split constituents was explored, and the methodology of cross validation for split constituents was studied to determine Astragalus sweet and warm property. The Astragalus was extracted by boiling water or other different solvents, and detected by HPLC-DAD or HPLC-ELSD. Finally, the similarity of each constituent was calculated by fingerprint software. Similarities of flavonoids and saponin constituents were all less than 0.31 and 0.34, respectively, compared to other constituents. The cross situation of nature-taste split components which was extracted by solvents was not serious. This method was proven to be feasible, and provided a theoretical and substantial basis for the Chinese taste pharmacological experiments and would be conductive to determine Astragalus sweet and warm property.
8.Potential mechanism of Huatan Qushi decoction on improving phlegm-dampness constitution using microRNA array and RT-qPCR targeting on hsa-miR-1237-3p
Zhang XIUPING ; Wang JI ; Liu BAOXING ; Yao HAIQIANG ; Chen YU ; Yin YUQING ; Yang XUEJING ; Li LINGRU
Journal of Traditional Chinese Medical Sciences 2021;8(1):43-51
Background: Metabolic diseases pose considerable burden on the healthcare system worldwide, indi-cating the significance of prevention and treatment. In constitution theory of traditional Chinese med-icine, phlegm-dampness constitution (PDC) is the common basis of metabolic diseases. In clinical practice, Huatan Qushi (HTQS) decoction targeting on PDC can effectively improve metabolic indicators. However, its underlying biochemical mechanism still remains unclear.Methods: Eight PDC participants received HTQS decoction for three months. Their blood was collected at baseline and 1 and 3 months after intervention started. Related biomedical indicators were detected. High-throughput sequencing and RT-qPCR were used for validation. Due to the missing data, repeated measures with missing values in mixed models were used. Results: After 3-month treatment, HDL-C level increased (P<.001) and FBG, FINS, and HbA1c all showed decreasing trend at different time points (all P < .05). After miRNA high-throughput sequencing, compared with the baseline, differential miRNAs at 1 and 3 months were screened, and target gene prediction and KEGG pathway enrichment analysis were performed. The results displayed that metabolic disease-related pathways mainly included pathways in cancer, PI3K-Akt signaling pathway, etc. Further, RT-qPCR showed that hsa-miR-1237-3p differed statistically (P =.008). Then we validated the target genes of hsa-miR-1237-3p in the"Pathways in Cancer"pathway including SDF1, AC, CRK, and HGF, also known as upstream target genes of PI3K/AKT pathway. The results showed that two indicators of CRK and HGF were in statistical significance (P=.045 and P=.036, respectively). Conclusion: PDC serves as a common basis for various metabolic diseases. Through adjusting PDC, HTQS decoction can improve biomedical indicators including blood glucose, HbA1c, insulin, and HDL-C. The target pathway is"Pathways in cancer". Specifically, HTQS decoction acts on targets of CRK and HGF by regulating hsa-miR-1237-3p, and probably exerts effects on their downstream PI3K/AKT pathway.
9.Prevalence of dyslipidemia and its influencing factors in rural residents aged 40 years and over in Liaoning Province
JING Li ; ZHANG Boqiang ; TIAN Yuanmeng ; YAN Han ; JIANG Haiqiang ; LIU Da ; LIU Shuang ; LIN Min ; YANG Zuosen ; XING Liying
Journal of Preventive Medicine 2020;32(5):449-454
Objective:
To understand the prevalence and influencing factors of dyslipidemia in the residents aged 40 years and over in the rural areas of Liaoning Province,so as to provide basis for the development of targeted prevention and control measures.
Methods:
From September 2017 to May 2018,by stratified cluster random sampling method,the residents aged 40 years or above from 19 villages in Liaoning Province were selected. Demographic features,height,weight,blood pressure and lipid level were collected. A logistic regression model was applied to explore the influencing factors for dyslipidemia.
Results:
A total of 10 926 residents were recruited,with an average age of (59.97±10.08)years. The crude and standardized prevalence rate of dyslipidemia was 30.96% and 29.68%. The results of multivariate logistic regression analysis showed that women(OR=1.323,95%CI:1.189-1.473),50-69 years old(OR:1.238-1.333,95%CI:1.075-1.523),a high school education or below(OR:0.585-0.635,95%CI:0.439-0.842),hypertension(OR=1.398,95%CI:1.273-1.534),diabetes(OR=2.137,95%CI:1.918-2.381),overweight or obesity(OR=2.101,95%CI:1.916-2.303), meat-based meals(OR=1.306,95%CI:1.144-1.492)and vegetables intake less than 5 days a week(OR:1.169-1.387,95%CI:1.004-1.796) were associated with dyslipidemiais.
Conclusions
The prevalence rate of dyslipidemia was 30.96% in the rural residents aged 40 years and over in Liaoning Province. People who were females,who were 50-69 years old,and who suffered from hypertension,diabetes,overweight or obese,might take their lipid levels into consideration.
10.Expression, purification and the biological activites of C-terminal fragment CTFβ
Linlin SONG ; Haiqiang YANG ; Ying ZHANG ; Jinsheng HE ; Yingcai GU ; Mingxu HOU ; Nannan LIU
Chinese Journal of Experimental and Clinical Virology 2014;28(5):395-397
Objective Proteolysis of the C-terminal fragment (CTFβ) of the amyloid precursor protein (APP) generates the Aβ peptides associated with Alzheimer' s disease (AD).The metabolism of CTFβ may play key roles in early stage of AD before Aβ generation.The aim of this study was to express,identify and purify the CTFβ,so as to provide evidence for its application in the development of AD detection system.Methods APP gene was used as the template,and the gene of CTFβ was cloned to pMD18-T vector through PCR.After sequencing,the CTFβ gene was cloned into the expression vector pET-30a(+) to construct the recombinant expression plasmid pET30a-CTFβ.The expression plasmid was transformed into Escherichia coli BL21 and the expression of CTFβ was induced by Isopropyl-β-D-thiogalactoside (IPTG).The effect of expression was confirmed by Western blottng.The recombinant protein was purified by using Ni-NTA affinity chromatography column,and the immunoreactivity of recombinant protein was detected by Western blotting and indirect ELISA.Results Western Blot results showed that recombinant protein was solubly expressed in E.coli and its molecular weight was about 10 × 103 to 25 × 103,and the size of fusion protein was consistent with prediction.In addition,the data of Western blotting showed that there were still some thick bands above the position of 80 × 103.Furthermore,the immunoblotting demonstrated that the 10× 103 to 25 × 103 of monomer of fusion protein was recognized by anti-histidine (his) tag and anti-Aβ (17-24) (4G8) antibody,while the high molecular aggregates were above 80 × 103 which were detected respectivly by anti-his,anti-Aβ antibody NU1,NU4 and A8.However,our data suggested that the bands above 80 × 103 were poorly recognized by fibril specific antibody NU6.These results demonstrated that the purified recombinant protein showed a specific immunoreactivity.Finally,indirect ELISA showed that the optimal concentration of CTFβ to coat the ELISA plate was 1 ng/well when it was used to detect Aβ antibody.Conclusion In this study,one of the aggregate-prone fragment of APP,CTFβ,was successfully expressed,purified,and identified,which would provide experimental clue to futher application in biochemical diagnosis of AD.