1.OCT and IVUS evaluating stent apposition and endothelialization after FD implantation in aneurysm animal models
Ji MA ; Shuhai LONG ; Jie YANG ; Zhen LI ; Haiqiang SANG ; Yi TANG ; Yuncai RAN ; Yong ZHANG ; Baohong WEN ; Shanshan XIE ; Ke CHEN ; Enjie LIU ; Xinwei HAN ; Tengfei LI
Chinese Journal of Neuromedicine 2024;23(3):256-262
Objective:To investigate the application value of optical coherence tomography (OCT) and intravascular ultrasound (IVUS) in evaluating flow diverter (FD) apposition and endothelialization in aneurysm animal models, and analyze the effect of incomplete stent apposition (ISA) on aneurysm lumen healing and stent endothelialization.Methods:Lateral common carotid artery aneurysm models in swines were established by surgical method and then FD was implanted. Immediately after surgery, OCT and IVUS were used to evaluate the locations and degrees of ISA, and difference between these 2 methods in evaluating FD apposition was compared. DSA was performed at 12 weeks after surgery to evaluate the aneurysm occlusion (Kamran grading) and stent patency. OCT and IVUS were used again to observe the stent endothelial situation; by comparing with histopathologic results, effect of ISA on aneurysm healing and stent endothelialization was analyzed.Results:Lateral common carotid artery aneurysm models in 6 swines were established, and 6 Tubridge FDs were successfully implanted. Compared with IVUS (3 stents, 4 locus), OCT could detect more ISA (6 stents, 14 locus); and the vascular diameter change area (7 locus), aneurysm neck area (4 locus) and the head and tail of FD (3 locus) were the main sites of FD malapposition; average distance between stent wire and vessel wall was (560.14±101.48) μm. At 12 weeks after surgery, DSA showed that 1 patient had a little residual contrast agent at the aneurysm neck (Kamran grading 3), and the remaining 5 had complete aneurysm occlusion (Kamran grading 4). One FD had moderate lumen stenosis, and the other 5 FDs had lumen patency. OCT indicated mostly disappeared acute ISA; ISA proportion decreased to 21.4 % (3/14), including 2 in the aneurysm neck and 1 in the partial stent. Histopathological results showed bare stent woven silk, without obvious endothelial coverage; in one FD with luminal stenosis, intimal hyperplasia was mainly composed of vascular smooth muscle cells.Conclusion:In carotid artery aneurysm model with FD implantation, OCT can detect more ISA than IVUS; most acute ISA have good outcome at 12 th week of follow-up, while severe ISA can cause delayed FD endothelialization and delayed aneurysm occlusion.
2.Effect of reperfusion injury following different ischemic duration on skeletal muscle in rats
Shengye ZHANG ; Linjie YANG ; Yunpeng LI ; Fudong HU ; Shengcun GUO ; Dong CHENG ; Yi TANG ; Ding YU ; Haiqiang SANG
Chinese Journal of Trauma 2022;38(2):172-181
Objective:To investigate effect of reperfusion injury following different ischemic duration on skeletal muscle in rats.Methods:A model of ischemia/reperfusion injury (IRI) was established by unilateral clamping femoral artery and additional application of tourniquet in skeletal muscle of hind limbs in 35 male Wsitar rats. According to different ischemia time, the animals were assigned to 2-hour ischemia and 24-hour reperfusion (I2R24 group), 2.5-hour ischemia and 24-hour reperfusion (I2.5R24 group), 3-hour ischemia and 24-hour reperfusion (I3R24 group), 4-hour ischemia and 24-hour reperfusion (I4R24 group) and sham group, with 7 rats per group. At the end of reperfusion, gastrocnemious tissues and plasma samples were collected and analyzed. The ratio of wet ∶ dry weight (W/D) was used to measure muscle edema. The assay of 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) was conducted to evaluate muscle viability. HE staining was executed to observe histopathological changes. Immunofluorescence staining was performed to assess the levels of C1q, C3b/c, tissue factor (TF), fibrinogen (FN), bradykinin receptor 1 (BR1), BR2, vascular cell adhesion molecule-1 (VCAM-1), E-selectin, fibrinogen-like protein-2 (FGL-2) and myeloperoxidase (MPO) in muscle tissues. ELISA method was used to determine the concentrations of interferonγ (IFN-γ), interleukin7 (IL-7), IL-18, macrophage inflammatory1α (MIP-1α) and monocyte chemotactic protein 1 (MCP-1) in plasma.Results:With prolongation of ischemia time and subsequent reperfusion, tissue edema became severe gradually. The ratio of W/D was 5.3±0.2, 6.1±0.3, 6.9±0.2, 7.6±0.3 in I2R24, I2.5R24, I3R24 and I4R24 groups, higher than that in sham group (4.5±0.1) (all P<0.01). Muscle viability got decreased gradually. Muscle viability was (62.4±3.5)%, (45.3±3.3)%, (35.4±3.4)%, (27.1±5.9)% in I2R24, I2.5R24, I3R24 and I4R24 groups, lower than that in sham group[(93.8±7.2)%](all P<0.01). Histopathological changes became aggravated gradually. The most severe group was I4R24 group, with the most severe myocyte injury, interstitial edema and extensive inflammatory infiltration, followed by I3R24, I2.5R24 and I2R24 groups in order. There was normal structure integrity and neatly arranged myocyte in sham group. Meanwhile, levels of C1q, C3b, FN, BR1, VCAM-1, E-selectin and FGL-2 got increased gradually. The highest levels for these factors were seen in I4R24 group, followed by I3R24 group, I2.5R24 group, I2R24 group and sham group in order. The rough ratio of the number of positive MPO cells/total cell number under high lens (×200) were increased gradually, with the highest level in I4R24 group, followed by I3R24 group, I2.5R24 group, I2R24 group and sham group in order. However, expression of TF and BR2 were not altered significantly among the groups. Plasma levels of INF-γ, IL-7, IL-18, MIP-1α and MCP-1 elevated gradually with prolongation of ischemia time (all P<0.01). The sequence was the sham group, I2R24 group, I2.5R24 group, I3R24 group and I4R24 groups for levels of these factors from low to high (all P<0.01). Conclusion:Reperfusion after prolongation of ischemia duration can increase the activation of complement, coagulation, kinin and endothelial cells as well as the release of inflammatory factors, and thus aggravate the degree of skeletal muscle tissue injury.
3.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.
4.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.
5.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 .
6.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.
7.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.
8.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.
9.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.
10.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.


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