1.Preparation of apolipoprotein M monoantibody and its preliminary application
Min HU ; Shuiping ZHAO ; Tao ZHANG ; Fangyan TIAN
Chinese Journal of Laboratory Medicine 2010;33(6):497-501
Objective To prepare anti-apoM monoantibodies with high affinity and high purity, and investigate apoM distribution among human tissues and different groups of people. Methods BALB/c mice were injected intracutaneously with recombinant apoM. After cytomixis, screening and cloning, we established a hybridoma, which grew well and steadily secreted antibodies. The ascites were acquired by injecting BALB/c mice intraperitoneally and anti-apoM monoantibodies were gained using standard techniques. We detected apoM levels in healthy individuals and the patients with coronary heart disease including stable angina (SA) group and acute coronary syndrome (ACS) group using the anti-apoM monoantibodies. ResultsThree anti-apoM monoantibodies were collected and confirmed after subtype identification and block test. The apoM protein were detected in some human cells and human tissues by these three monoantibodies. The concentration of apoM was (11.02 ±1.96) ×10 -3 g/L, (10. 76± 1.32) ×10-3 g/L, (12. 83 ± 2. 28) × 10-3 g/L in SA, ACS and control group respectively. There was significant difference within the three groups (F = 11. 544, P < 0. 05). Comparing apoM concentrations among control group and coronary heart disease groups, it showed that the levels of apoM were lower in coronary heart disease groups than in control group(t =2. 962 and 3. 967,P <0. 05). There was no significant difference between two coronary heart disease groups (t = 1. 033, P > 0. 05). Conclusion Anti-apoM monoantibodies are successfully raised and could combine with apoM in human cells and tissues. This lays the foundation for the apoM study in apolipoprotein metabolism.
2.Effects of Extractive from Gastrodiae Rhizoma on Learning-Memory Function in Mice
Chunni HAN ; Fangyan HE ; Ye TIAN ; Xiaohua DUAN
Chinese Journal of Information on Traditional Chinese Medicine 2014;(9):50-52
Objective To evaluate the effects of extractive from Gastrodiae Rhizoma on acquisition, consolidation and retrieval of learning-memory function in mice;To provide some reference for clinical research and development of new drugs.Methods Male Kunming mice were randomly divided into control group, model group, positive control group and Gastrodia extractive group. Positive control group and Gastrodia extractive group were given gavage by using relevant medicine 0.2 mL/10 g, and the control group and model group were given gavage with the same amount of distilled water for 16 days. After receiving gavage for continuous 11 days, memory acquisition barrier model was induced by scopolamine;memory consolidation barrier model was induced by chloromycetin;memory retrieval barrier model was induced by EtOH. The learning-memory function was reviewed by escape latency and spatial search distance. The quadrant and distance search time percentage was detected through directional navigation test and spatial probe test in Morris water maze.Results Extractive from Gastrodia Rhizoma shortened the time for acquisition, consolidation and retrieval of learning memory about escape latency and spatial search distance (P<0.05,P<0.01), and the quadrant and distance search time percentage were prolonged (P<0.01).Conclusion Extractive from Gastrodia Rhizoa can effectively improve the acquisition, consolidation and retrieval of learning-memory function in mice.
3.The relationship between three-dimensional right ventricular longitudinal deformation and myocardial fibrosis in patients with end-stage heart failure
Fangyan TIAN ; Yuman LI ; Yanting ZHANG ; Ying GU ; Bei ZHANG ; Mingxing XIE
Chinese Journal of Ultrasonography 2021;30(5):376-381
Objective:To investigate the values of right ventricular free wall longitudinal strain (RVFWLS) by three-dimensional speckle tracking echocardiography (3D-STE) in predicting the degree of RV myocardial fibrosis (MF) in patients with end-stage heart failure (HF).Methods:A total of 102 consecutive patients with end-stage HF undergoing heart transplantation were enrolled in the Union Hospital of Tongji Medical College, Huazhong University of Science and Technology from June 2018 to December 2019. Echocardiographic examinations were performed in these patients before heart transplantation. The conventional RV function parameters were obtained, including fractional area change, tricuspid annular plane systolic excursion(TAPSE), myocardial performance index, tricuspid lateral annular systolic velocity(Tricuspid s′). Two-dimensional (2D) RVFWLS was calculated by two-dimensional speckle tracking echocardiography (2D-STE). Right ventricular (RV) end-diastolic volume (RVEDV), RV end-systolic volume (RVESV), RV stroke volume (RVSV), RV ejection fraction (RVEF) and 3D-RVFWLS were measured by 3D-STE. The degree of MF was quantified using Masson′s trichrome stain in RV myocardial samples after heart transplantation. Patients were divided into mild, moderate, and severe groups according to the degree of MF on histology, then echocardiographic parameters were compared among the 3 groups. Pearson correlation analysis and the multiple linear regression analysis between echocardiographic parameters and RV MF were analyzed.Results:Compared with patients with mild and moderate MF, 3D-RVFWLS, 2D-RVFWLS and conventional parameters of RV function were significantly decreased in patients with severe MF.RV MF strongly correlated with 3D-RVFWLS ( r=-0.71, P<0.01), modestly correlated with 2D-RVFWLS ( r=-0.53, P<0.01), and weakly correlated with RVFAC, TAPSE, RVEF, Tricuspid s′, RVSV ( r=-0.47, -0.44, -0.35, -0.29, -0.38; all P<0.01). 3D-RVFWLS correlated best with the degree of MF compared with 2D-RVFWLS and conventional RV function parameters ( r=-0.71 vs r=-0.29~-0.53, all P<0.05). A stepwise multivariate analysis showed that 3D-RVFWLS was independently associated with RV MF (β=1.554, P<0.01, adjusted R2=0.539). Conclusions:3D-RVFWLS can provide an important imaging reference for detecting the degree of RV MF in patients with end-stage HF.
4.Effect of dynamics of instantaneous flow rate on the quantification of the severity of degenerative mitral regurgitation using M-mode proximal isovelocity surface area
Chunqiang HU ; Zhenyi GE ; Shihai ZHAO ; Fangyan TIAN ; Wei LI ; Lili DONG ; Yongshi WANG ; Dehong KONG ; Fangmin MENG ; Zhengdan GE ; Xianhong SHU ; Cuizhen PAN
Chinese Journal of Ultrasonography 2023;32(7):590-599
Objective:To investigate the effect of instantaneous flow rate on the consistency of diagnostic accuracy of severe degenerative mitral regurgitation (DMR) using proximal isovelocity surface area (PISA).Methods:From June 2019 to June 2021, 75 patients with DMR who underwent echocardiography in Department of Echocardiography of Zhongshan Hospital, Fudan University were prospectively enrolled. The instantaneous flow rate of DMR during the systolic phase was calculated using M-mode PISA(PISA M-mode), and a time-integrated curve was plotted. Regurgitant volume (RVol) and effective regurgitant orifice area (EROA) were calculated by traditional PISA (PISA max), pair PISA (PISA pair), and PISA M-mode, respectively. RVol acquired from cardiac magnetic resonance (CMR) volumetric method in 22 patients of the enrolled patients. The correlation and consistency of RVol acquired between the three PISA methods and CMR were compared. Agreement of diagnostic accuracy of severe mitral regurgitation (sMR) acquired between the three PISA methods and multi-parameter algorithm by American Society of Echocardiography (ASE) was analyzed using Cohen′s Kappa analysis. Results:The curve of instantaneous flow rate of DMR showed unimodal pattern with the peak at mid-late systolic phase. The correlation of RVol acquired between PISA methods and CMR was moderate for PISA max and PISA pair ( r=0.77, 0.80, both P<0.001), whereas PISA M-mode presented strong correlation with CMR ( r=0.87, P<0.001). RVol acquired from PISA max was larger than that of CMR[(69.1±37.1) ml vs (49.0±29.0)ml, P=0.002]. Both PISA max and PISA pair were shown moderate agreement of diagnostic accuracy of sMR with ASE multi-parameters algorithm (RVol: κ=0.496, 0.525, both P<0.001; EROA: κ=0.570, 0.578, both P<0.001), while PISA M-mode presented strong agreement (RVol: κ=0.867 and EROA: κ=0.802, both P<0.001). Conclusions:Based on the unimodal pattern of instantaneous flow rate in patients with DMR, PISA max may significantly overestimate RVol, exposing a significant proportion of patients with DMR to unnecessary MR surgery. PISA M-mode presents better correlation and consistency with CMR on the quantification of RVol compared with PISA max and PISA pair, and may improve the diagnostic accuracy of quantification of sMR using PISA.
5.Optimization of the Best Proportion of Astragalus membranaceus Injection Combined with Erigeron breviscapus Injection against Cerebral Ischemia-reperfusion Injury in Rats by Baseline Geometric Proportion Increasing and Decreasing Design
Feng TIAN ; Jicao LI ; Shaowei TANG ; Yushi SUN ; Shijia LIN ; Fangyan ZHANG ; Xingliang XIE ; Yanmei SHENG
China Pharmacy 2019;30(14):1885-1889
OBJECTIVE: To screen the best proportion of Astragalus membranaceus injection combined with Erigeron breviscapus injection against cerebral ischemia-reperfusion injury in rats. METHODS: Male SD rats were randomly divided into sham operation group, model group and administration group [different A. membranaceus injection-E. breviscapus injection proportion groups, being A(0 ∶ 10), B(2 ∶ 8), C(4 ∶ 6), D(6 ∶ 4), E(8 ∶ 2), F(10 ∶ 0)groups, set by baseline geometric proportion increasing and decreasing design], with 8 rats in each group. Except for sham operation group, reperfusion injury model of middle cerebral artery occlusion were induced by modified suture method in rats. The each administration group was given relevant medicine intraperitoneally once immediately after inducing model, and then given again after 24 hours (medication interval between the two injections of 30 min). Constant volume of normal saline was given to rats in sham operation group and model group. Forty-eight hours after reperfusion, Longa scoring method was used to evaluate neurological impairment of rats, and neurological impairment score was recorded. Serum content of MDA and activity of SOD were measured by colorimetry assay. TTC assay was used to detect cerebral infraction, and cerebral infarction rate was calculated. Kim’s formula was used to calculate the synergistic index (q) of rats in administration groups. RESULTS: Compared with sham operation group, neurological impairment score and serum content of MDA were increased significantly in model group, while activity of SOD was decreased significantly (P<0.01). The area of cerebral infarction increased significantly, and the rate of cerebral infarction increased significantly (P<0.01). Compared with model group, neurological impairment scores and serum contents of MDA were decreased significantly in group A, B, C, D and E; neurological impairment score of group C was significantly lower than those of group A and F; serum contents of MDA in group B, C, D and E were significantly lower than that of group F (P<0.05 or P<0.01). Activities of SOD in group A, B, C, D and E were increased significantly, and group C was significantly higher than group F (P<0.05 or P<0.01). The cerebral infarction area of rats in each administration group was reduced to varying degrees. The cerebral infarction rates of rats in group B, C, D and E were significantly reduced, and group C and D were significantly lower than group F, while group C was significantly lower than group A (P<0.05 or P<0.01). The q values of group B, C, D and E were 0.90, 1.30, 1.00, 0.70 (neurological impairment score) and 0.79, 1.27, 0.98, 0.82 (cerebral infarction rate). CONCLUSIONS: Different ratios of A. membranaceus injection and E. breviscapus injection have certain protective effects on cerebral ischemia-reperfusion injury model rats, can relieve their neurological deficits, alleviate their oxidative stress and reduce their cerebral infarction areas. The effect of the combination of the two drugs is better than that of single use, and the optimum ratio is 4 ∶ 6.