1.Determination of Angelica Sinensis Polysaccharide ( ASP1 ) in Rats by HPLC-FD with Pre-column Derivat-ization
China Pharmacist 2017;20(3):438-442
Objective:To establish an HPLC-FD method for determining the content of Angelica sinensis polysaccharide (ASP1) to lay foundation for its pharmacokinetic study in rats. Methods: Purified ASP1 was labeled with FITC by the method of Belder and Granath to obtain ASP1-FITC. The tissue samples were treated with 30% trichloroacetic acid and 11% NaOH before injection. The samples were determined by HPLC-FD. A PL aquagel-OH MIXED column was used,and the mobile phase was phosphate buffer( dis-solve NaH2PO32. 34g , Na2HPO34. 33 g and NaCl 11. 70 g into 1000 ml water) with pH of 7. 0. The flow rate was 0. 5 ml·min-1. The excitation and emission wavelengths was set at 495 nm and 520 nm, respectively. Results:The linear calibration curve was within the concentration range of 0. 25-40. 00 μg·ml-1(r=0. 9996)with the lower limit of quantification of 0. 20μg·ml-1 in tissue sam-ples. The extraction recovery of ASP1 was determined at low, medium and high concentration with the recovery of 91. 98%-114. 20%. The intra and inter-day RSDs were lower than 8. 31% and 2. 94%, respectively. Conclusion:The method to determine the content of ASP1 in rats by HPLC-FD with pre-column derivatization has been esablished. It is simple, accurate and reliable, which can be suc-cessfully applied in the study of pharmacokinetics and tissue distribution of ASP1 in rats.
2.The study of heat shock protein 70 as a adjuvant carrier on HBV-specific immune response in vitro
Li LUO ; Song HE ; Na LUO ; Mingli PENG
Chongqing Medicine 2014;(9):1025-1028
Objective To investigate the effect of mycobacterium tuberculosis heat shock protein 70(TB .HSP70) as an adjuvant carrier on stimulating hepatitis B virus (HBV) core antigen(HBcAg)specific immune response to an accompanying cytotoxic T lym-phocytes epitope peptide from HBV core antigen in vitro .Methods Recombinant proteins HSP70(P1)、HSP70-HBcAg(18-27) (P2)、HSP70-PreS2B (18-24)-PreS2Th(37-53)-HBcAg(18-27)(P3) were expressed in methylotropic yeast Pichia pastoris GS115 . The expression of recombinant proteins was identified by SDS-PAGE and Western blot .The effect of recombinant proteins on den-dritic cell and lymphocytes of chronic HBV infection volunteers was investigated in vitro .The maturation of dendritic cell was meas-ured by flow cytometry ;the secretion of Th1 cytokines such as IL-12p70 ,IL-1β,TNF-αand IFN-γ was measured by ELISA ;the proliferation of lymphocytes was measured by TdR-3H ;the HBV-spesific cytotoxic activity was measured by the classic 51 Cr .Re-sults The recombinant proteins (P1 ,P2 ,P3) were constructed successfully .P1 ,P2 ,P3 could activate dendritic cell from chronic HBV infection volunteers by upregulation CD1a ,CD40 ,CD86 and production Th1 cytokines such as IL-12p70 ,IL-1β and TNF-α. Especially P3 could better induce autologous T cells to generate HBV specific cytotoxic T lymphocytes response ,activate the prolif-eration of lymphocytes and release IFN-γeffectively .However ,the recombinant HSP70 showed no target cell killing and could not induce immune response effectively .Conclusion TB .HSP70 can be used as an adjuvant carrier to stimulate HBV specific immune response to an accompanying cytotoxic T lymphocytes epitope peptide from HBV core antigen ,and enhance immunogenicity of the cytotoxic T lymphocytes epitope peptide .The P3 with B-and T-epitope can activate the HBV specific immune response effectively .
3.Discussion on circulating fluid replacement in the polyuria stage after simultaneous islet cell-renal transplantation
Li DONG ; Hong LI ; Na LI ; Zhifeng LUO
Chinese Journal of Practical Nursing 2013;29(29):41-43
Objective To discuss and improve the scheme of circulating fluid replacement in the polyuria stage after simultaneous islet cell-renal transplantation.Methods Nursing data of 26 patients after simultaneous islet cell-renal transplantation during the polyuria stage were analyzed retrospectively,the impact on spend-and-tax circulating fluid replacement on the outcome after surgery was observed.Results In 26 patients,3 case suffered graft function recovery delay,23 patients' renal function recovered smoothly without water-electrolyte and acid base disorders in period of diuresis,compared with the 16 cases with traditional fluid replacement in the same time,serum creatinine (SCr),blood urea nitrogen (BUN) and blood glucose (BG) decreased significantly in 72 h.Conclusions Using spend-and-tax fluid replacement can reduce postoperative complications such as water-electrolyte and acid-base disorders in simultaneous islet cell-renal transplantation patients,and promote the functional recovery of transplanted islet cells and kidney.
4.Investigation on autologous coronary intervention in patients with angina recurrence of graft vessels occlusion after coronary artery bypass graftting
Chengyang LI ; Zhilin MIAO ; Zhanquan LI ; Na DUAN ; Defeng LUO
Chinese Journal of Primary Medicine and Pharmacy 2011;18(z1):1-2
Objective To investigate the effect of autologous coronary intervention in patients with angina recurrence of graft vessels occlusion after coronary artery bypass graftting(CABG).Methods Retrospectively analyzed the data of 10 patients with angina recurrence because of graft vessels occlusion.treated by CABG,including in clinical data,arteriography and the interventional results.Results Among 10 patients,9 patients received chronic total occlusion(CTO) PCI,another 1 patients received left main stem(LM) intervention.There were none had angina recurrence after PCI in 10 patients.Conclusion Conclusion Autologous coronary intervention in patients with angina recurrence of graft vessels occlusion after coronary artery bypass graftting was the safety and effective treatment.
5.A pilot study on photoreceptor IS/OS and subfoveal choroidal thickness in IMH with spectral domain OCT
Zefeng, XIAO ; Na, LUO ; Fabiao, LI ; Zhiqiao, QIU ; Shigang, YAN
Chinese Journal of Experimental Ophthalmology 2015;33(11):1019-1022
Background The classical theory from Gass holds that idiopathic macular hole (IMH) is due to the dragging of vitreous cortex in the tangent direction on the macular region,while spectral domain OCT (SD-OCT)studies found that the diameters of inner segment/outer segment (IS/OS) absence (DIOA) region was closely associated with vision ability of IMH patients.Enhanced depth imaging (EDI) is thought to be a new method to study the relation between DIOA and subfoveal choroidal thickness (SFCT) ,but there are few relevant reports.Objective This study aimed to observe the correlation between photoreceptor IS/OS junction and SFCT before and after vitrectomy in IMH by EDI SD-OCT.Methods Forty unilateral IMH patients were enrolled in Affiliated Foshan Hospital of Southern Medical University from June 2011 to June 2013.Vitrectomy (23G) was performed on the patients by the same operator.EDI mode of SD-OCT was used to measure DIOA and SFCT in horizontal direction before and 6 months after surgery.The relationship between DIOA and SFCT was evaluated using Pearson linear correlation analysis.Written informed consent was obtained from each patient before medical assessment.Results IMH was anatomically closed after vitrectomy in 34 eyes with the closure rate 85%.The mean DIOA was (1 280±753) μm in preoperation, which was significantly higher than (656 ± 322) μm in postoperation (t =4.989, P =0.000).The mean SFCT was (130±43)μm in preoperation,showing a slight reduce in comparison with (140±38)μrn in postoperation, but there was not statistical significance between them (t =-1.407, P =0.175).Negative correlations were found between DIOA and SFCT in both preoperation and postoperation (preoperation:r=-0.748 ,P=0.000;postoperation: r=-0.686,P =0.001).Conclusions The alteration of DIOA shows a negative correlation with SFCT in IMH eyes,suggesting a contributing role of the choroid in the change of photoreceptors of IMH.
7.Investigation and prevention of dermatosis affecting pilots
Guang ZHAO ; Fei LI ; Aihua NA ; Wei LUO
Medical Journal of Chinese People's Liberation Army 2001;0(11):-
Objective To investigate the prevalence of dermatosis affecting pilots in different areas. Methods From Oct. 2007 to Sep. 2008,epidemiological investigation and clinical examination were carried out in 255 pilots stationed at Beijing,Tianjin,Hubei and Shaanxi regions. In order to get the correct diagnosis of dermatosis and the accurate number of infection cases/time,same medication and management were applied for the treatment of those skin disorders. The epideomology of dermatosis in pilots of different troops and age groups were then compared,and the correlation factors affecting the epidemics were analyzed with Chi-square test. Results Among 255 pilots,430 cases (times) were found to suffer from 47 kinds of dermatosis,of whom 72% suffered from two or more kinds of dermatosis. The incidence of superficial mycosis (69.8%) took ranked the highest. The incidence of dermatosis was different among the pilots stationed in different regions. The prevalence of lichen simplex chronicus was higher in the pilots stationed in Beijing,Tianjin and Shaanxi regions than in those in Hubei region,while the prevalence of tinea manuum and tinea pedis showed the opposite status (P 7 year enlistment (P
8.THE INFLUENCING ELEMENTS AND OPTIMIZATION OF X-gal STAINING METHOD IN THE STUDY OF TRANSPLANTATION
Qi ZHANG ; Jinhong MENG ; Na LUO ; Jingwen LI ; Gong JU
Acta Anatomica Sinica 1955;0(03):-
Objective To study the influencing elements in x-Gal staining method and optimize the reactive conditions so that nonspecific background can be eliminated and grafted exogenous cells carrying LacZ gene can be discerned correctly. Methods C17 2 cells (carrying lzcZ gene) were injected into the right lateral ventricle both in the adult and newborn animals. After one week they were perfused using two methods, then the slices were stained at different pH and incubating time respectively. The X-gal positive cells in hippocampus were counted under light microscope. Results Background staining in this method has close correlation with the species and age of the host animals and it decreases when pH is higher or incubating time is shorter. The results are the best when pH 9 5 and the incubating time is 1*!h.Conclusion The reliability of X-gal staining method depends on optimization of several parameters, including pH, incubating time, perfusion etc. It is necessary to establish the correspondent controls.;
9.Effect of NF-?B inhibitor on renal expression of apolipoprotein M in rats with acute renal failure
Hua TANG ; Zhuhua LI ; Ping ZOU ; Na LUO ; Chunmei ZHANG
Chinese Journal of Pathophysiology 2000;0(11):-
AIM:To investigate the expression pattern of apolipoprotein M (apoM) protein in renal cortex of acute renal failure (ARF) rats with reperfusion. METHODS:Seventy-five male rats were randomly divided into sham operation group (n=25),ARF group (n=25) and pyrrolidine dithiocarbamat (PDTC) group (n=25),five subgroups at time points of 3 h,6 h,12 h,24 h and 48 h after reperfusion were set up in each group. The expressions of apoM in cytoplasm and NF-?B p65 in nucleus of renal cortex were detected at the indicated time points. RESULTS:The expression of apoM in ARF group was obviously higher than that in sham operation group (P
10.Effect of NF - κB inhibitor on renal expression of apolipoprotein M in rats with acute renal failure
Hua TANG ; Zhuhua LI ; Ping ZOU ; Na LUO ; Chunmei ZHANG
Chinese Journal of Pathophysiology 2009;25(11):2141-2144
AIM: To investigate the expression pattern of apolipoprotein M (apoM) protein in renal cortex of a-cute renal failure ( ARF) rats with reperfusion. METHODS: Seventy - five male rats were randomly divided into sham operation group (re =25) , ARF group (n =25) and pyrrolidine dithiocarbamat (PDTC) group (n =25) , five subgroups at time points of 3 h, 6 h, 12 h, 24 h and 48 h after reperfusion were set up in each group. The expressions of apoM in cytoplasm and NF - κB p65 in nucleus of renal cortex were detected at the indicated time points. RESULTS: The expression of apoM in ARF group was obviously higher than that in sham operation group ( P <0.01 ) , and two peaks were detected, the first peak was at 6 h after reperfusion, while the second one was from 24 h to 48 h. The tendency of apoM expression in PDTC group was similar to that in ARF group, while the expression in every subgroup was prevalently lower than that in ARF group (P < 0.01). Otherwise, a significant correlation ( r = 0.852, P < 0.01) was found between the expression of apoM and NF -κB p65.CONCLUSION: The results indicate that apoM feasibly take part in the pathogenesis of ARF through the inflammatory reaction mediated by NF - κB.