1.The exposure dose-effectiveness of ? irradiation on killing the plasmodium in mice red blood cells
Zhongxiang LIU ; Yinghui LI ; Ya ZHAO
Chinese Journal of Blood Transfusion 2001;0(06):-
Objective To study the exposure dose-effectiveness of ? irradiation on killing the plasmodium in the mice RBC,for the further exploration on the method that could kill the plasmodium in RBC without affecting the activity and function of normal RBC. Methods After infection with Plamodium yoelii (P.y),blood was collected from mice and exposed to ? irradiation (radiated group). An unirradiated group served as control. In the irradiated group,P.y infected blood was divided into three aliquots,each aliquot was irradiated one time by ? radiation using Gammacell 1000 Elite blood radiation apparatus. The dosage of each aliquot was 25,35 and 45Gy. After irradiation,the blood samples were stored at 4℃. Then mice were inoculated with these irradiated blood stored for 1,3 or 5 days after irradiation,or with unirradiated blood. Two days later,the blood samples were taken from inoculated mice and were examined under microscope and plasmodium infection rates were calculated. Results The mice in the control group had parasitemia much earlier than those in irradiated group (1—2 days),and the plasmodium infection rate in the control group was significantly higher than that in the irradiated group(3.7% vs 0.07%). With increasing dosage of irradiation,the survived plasmodium in blood decreased,and survival of mice increased(8—12 days). After 45 Gy irradiation and 5 day storage at 4℃,there were no plasmodium found in the red blood cell of inoculated mice. In the control group,blood testing result was positive,and all the mice died.Conclusion Plasmodium in mice RBC can be killed effectively when blood is exposed to 45Gy irradiation and stored at 4℃ for 5 days.
2.Different sample collecting and template making methods in malarial PCR diagnosis
Zhongxiang LIU ; Minglin SUN ; Ya ZHAO
Chinese Journal of Laboratory Medicine 2001;0(01):-
Objective To find the best sample collecting and template making methods. Methods The multiplex PCR results of three sample collecting methods and eight template making methods in malaria diagnosis were compared. Results Conserved blood sample collecting, and Na 3PO 4 template making were sensitive and simple. Conclusion Conserved blood of sample collecting and Na 3PO 4 in template making are the best methods in multiplex PCR diagnosis of malaria, and are worthy of wide use.
3.Triterpenes from Callicarpa integerrima Champ.
Chenchen ZHU ; Li GAO ; Zhongxiang ZHAO ; Chaozhan LIN
Acta Pharmaceutica Sinica 2012;47(1):77-83
A new triterpenoid saponin and fourteen known triterpenoids were isolated from the methanol extract of the stems and leaves of Callicarpa integerrima Champ, which is used in Chinese folk medicine for stopping bleeding, expelling the wind, dissipating stagnation, and treating scrofula, by using various chromatographies, such as silica gel, Sephadex LH-20 and RP-C18 column chromatography. Their structures were identified as a new compound 2alpha, 3beta, 19alpha, 23-tetrahydroxy-olean-12-en-28-oic acid-28-O-beta-D-glucopyranosyl-(1 --> 4)-beta-D-glucopyranoside (1), together with fourteen known compounds: oleanolic acid (2), 3-acetyl oleanolic acid (3), 3beta-O-acetyl ursolic acid (4), 2alpha-hydroxy-ursolic acid (5), 2alpha, 3beta, 19alpha, 23-tetrahydroxy-urs-12-en-28-oic acid (6), alpha-amyrin-3-O-beta-D-glucopyranoside (7), pomolic acid (8), betulinic acid (9), ursolic acid (10), 2alpha, 3beta, 19alpha, 23-tetrahydroxy-olean-12-en-28-oic acid (arjungenin) (11), 2alpha-hydroxy-oleanolic acid (12), hederagenin (13), 2alpha, 19alpha-dihydroxy-ursolic acid (14) and pruvuloside A (15), by the spectroscopic techniques of NMR, HMBC, IR and MS, separately. All these compounds were obtained from this plant for the first time, and compounds 3, 4 and 15 were isolated from genus Callicarpa L. for the first time.
4.Simultaneous determination of seven flavonoids in Nervilia fordii with HPLC.
Li ZHANG ; Chenchen ZHU ; Zhongxiang ZHAO ; Chaozhan LIN
Acta Pharmaceutica Sinica 2011;46(10):1237-40
The study is to develop an HPLC method for simultaneous determination of rhamnazin (1), rhamnocitrin (2), rhamnetin (3), rhamnazin-3-O-beta-D-glucopyranoside (4), rhamnazin-3-O-beta-D-xylopyranosyl-(1-->4)-beta-D-glucopyranoside (5), rhamnazin-3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranoside (6), and rhamnocitrin-3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranoside (7) in Nervilia fordii. The separation was performed on a Kromasil C18 column (250 mm x 4.6 mm, 5 microm) with 0.4% phosphoric acid-acetonitrile as the mobile phase in a gradient elution at a flow rate of 1.0 mL x min(-1). The detect wavelength was set at 256 nm, and the column temperature was set at 40 degrees C. There were good linear relationships between the logarithm values of concentrations and those of the peak areas of seven flavonoids (1-7) in the range of 0.55-70.00 microg x mL(-1) (r = 0.9997), 0.86-110.00 microg x mL(-1) (r = 0.9997), 0.39-50.00 microg x mL(-1) (r = 0.999 7), 0.55-70.00 microg x mL(-1) (r = 0.999 5), 1.33-170.00 microg x mL(-1) (r = 0.9998), 1.33-170.00 microg x mL(-1) (r = 0.9998), 0.16-20.00 microg x mL(-1) (r = 0.9995), respectively. The recoveries of the seven flavonoids were between 97.19%-99.45%, the relative standard deviations (RSDs) were between 0.91%-2.69%. The established method is rapid, accurate with high repeatability, which could provide scientific evidence for the quality control of Nervilia fordii.
5.Tanshinone IIA protects against triptolide-induced liver injury via Nrf2/ARE activation.
Cuiwen GUAN ; Jing JIN ; Jia LI ; Zhongxiang ZHAO ; Zhiying HUANG
Acta Pharmaceutica Sinica 2013;48(9):1397-402
The aim of this study is to investigate the protection effect of tanshinone IIA (Tan) against triptolide (TP)-induced liver injury and the mechanisms involved. Acute liver injury was induced by intraperitoneal injection of TP (1 mg x kg(-1)) in mice. The activities of AST, ALT and LDH in serum and the levels of GSH, GST, GSH-PX, SOD, CAT and MDA in liver tissue were detected. The histopathological changes of liver tissues were observed after HE staining. Nrf2 translocation in liver tissue was detected by Western blotting, and real-time PCR was used to measure the expression levels of GCLC, NQO1 and HO-1 mRNA. The results showed that pretreatment with Tan significantly prevented the TP induced liver injury as indicated by reducing the activities of AST, ALT and LDH (P < 0.01). Tan pretreatment also prevented TP-induced oxidative stress in the mice liver by inhibiting MDA and restoring the levels of GSH, GST, SOD and CAT (P < 0.05). Parallel to these changes, pretreatment with Tan could attenuate histopathologic changes induced by TP. Furthermore, the results indicated that Tan pretreatment caused nuclear accumulation of Nrf2 as well as induction of mRNA expression of antioxidant response element (ARE)-driven genes such as GCLC, NQO1 and HO-1. These results indicated that Tan could protect against TP-induced acute liver injury via the activation of Nrf2/ARE pathway.
6.IN VIVO EFFECT OF 3′,5′-cAMP ON EHRLICH ASCITES TUMOR CELLS AND THE CHANGES OF THE INTRACELLULAR 3′,5′_cAMP,cAMP-PDE ACTIVITY AND THE CHANGES OF MICROVILLI IN THESE TUMOR CELLS
Daishu WANG ; Menglian ZHAO ; Zhongxiang LIN ; Yunyan LIANG ; Kunren WANG ;
Acta Anatomica Sinica 1953;0(01):-
This experiment was carried out in young adult albino mice,which were injectedwith Ehrlich ascites tumor cells intraperitoneally to produce Ehrlich ascites tumor.The animals were divided into 2 groups.The first group received cAMP plusaminophylline for 5 to 13 days after inoculation.The second group received saline ascontrol.We found that in the administration of cAMP together with aminophylline,thegrowth of Ehrlich ascites tumor cells was inhibited to the extent of 53% at the 9thdays after inoculation.Early or later than the 9th day,the inhibitory rates were marklylower.By using cAMP immunocytochemical method,it was found that on the9th day of inoculation,there was an increase of the intensity of intracellualr cAMPspecific fluorescence in tumor cells of the cAMP treated mice in comparison withthose of the control.It also inhibited the 3',5'-cAMP-PDE activity on the 5th to 7thday after inoculation.Under the dark field microscope on the surface of the Ehrlich tumor cells therewere numerous“brush-like”microvilli,but they were not visible on the surfaceafter treated with cAMP together with aminophylline.The agglutination by theCon.A,was decreased markly on the 7th to 9th day after the inoculation.The possible relationship between the level of intracellular cAMP and of the3',5'-cAMP-PDE,and especially the inhibition of the formation of microvilli onthe treated tumor cell surface is discussed in this paper.
7.Total triterpeniods from Rabdosia lophanthoides var.gerardianus with macro porous resin
Chaozhan LIN ; Cuixian ZHANG ; Zhongxiang ZHAO ; Chenchen ZHU
Chinese Traditional Patent Medicine 1992;0(01):-
AIM: To study the enrichment-purification process of total triterpeniods from Rabdosia lophanthoides var.gerardianus with macro porous resin. METHODS: The purify of the total triterpeniods was used as marker to optimize the adsorptive capacity and elution characteristics. RESULTS: The result showed that 20 mL of the extraction solution(0.5 g/mL) was purified with a column of macro porous resin(d15 mm?h120 mm,V=20 mL,dried weight 10 g) and washed with distilled water,then eluted with 80 mL 60% ethanol and 160 mL 90% ethanol in proper order.With macro reticular resin to adsorb and purify,the elution ratio of total triterpeniods of 90% ethanol fraction was 93.2% and the purity reached above 20%. CONCLUSION: This process of applying macro reticular resin to adsorbing and purifying total triterpeniods from Herba Rabdosia lophanthoides var.gerardianus was successful.
8.Study on Antioxidant Stilbenes from the Rhizomes of Smilax china
Zhongxiang ZHAO ; Jing JIN ; Chenchen ZHU ; Cuixian ZHANG ; Jinlan RUAN
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(02):-
Objective To study the stilbene constituents from the traditional Chinese medicine Smilax china and to determine their antioxidant activity. Methods The compounds were separated and purified by column chromatography with silica gel, RP C18, and Sephadex LH- 20, and were identified by IR, MS, NMR. DPPH method was used to evaluate the free radical scavenging activity of the isolated compounds. Results Three compounds were isolated from the EtOAc fraction of the rhizomes of S. china and were identified as: resveratrol (1), oxyresveratrol (2) and 3, 5, 3′ , 4′ - tetrahydroxylstilbene (3). Compounds 1~ 3 showed strong antioxidant activity, and could scavenge DPPH free radicals, effectively. At the concentration of 50 ? mol/L, their DPPH free radical scavenging rates were 79.47 % , 89.89 % and 93.86 % , respectively. Conclusion Stilbenes might be the material foundation of antioxidant activities of rhizomes of S. china.
9.Survey of marriage and mood of patients with spinal cord injury survived after Tangshan earthquake
Songhuai LIU ; Jianjun LI ; Hongjun ZHOU ; Zhongxiang MI ; Chaonan ZHAO
Chinese Journal of Rehabilitation Theory and Practice 2005;11(2):113-114
ObjectiveTo explore the effect of marriage on the life satisfaction and mood of the spinal cord injury couple survived after Tangshan earthquake.Methods40 SCI patients (20 married and 20 unmarried) who were survived after Tangshan earthquake were investigated with 20 from married family (10 male and 10 female) and 20 from unmarried family (10 male and 10 female). The contents of investigation included life satisfaction, quality of life, anxiety and depression.Results The married group had significantly more satisfied with their life and less anxiety than the unmarried group. But there were no differences in evaluation of life quality and depression between two groups.Conclusion The marriage can improve the life satisfaction and psychological health of the SCI patients survived after Tangshan earthquake.
10.Changes and significance of ubiquitination of Foxp3 protein during the acute phase of Kawasaki dis-ease
Junshan ZHAO ; Qin WANG ; Pengqiang WEN ; Mingguo XU ; Zhongxiang QI ; Chengrong LI ; Guobing WANG
Chinese Journal of Microbiology and Immunology 2015;(6):436-441
Objective To investigate the changes and significance of ubiquitination of Foxp3 pro-tein during the acute phage of Kawasaki disease ( KD) .Methods Forty-eight children with KD and twenty-eight age-matched healthy children were recruited in this study.Co-immunoprecipitation and Western blot assays were performed to determine the poly-ubiquitination status of Foxp3 in CD4+T cells.The percentages of CD4+CD25high Foxp3+regulatory T cells ( Treg) and the levels of IL-10, transforming growth factor-β( TGF-β) , cytotoxic T lymphocyte-associated protein-4 ( CTLA4 ) , STIP1 homology and U-Box containing protein 1 (STUB1), heat shock protein 70 (HSP70), ubiquitin-specific-processing protease 7 (USP7) and pSTAT3 were analyzed by flow cytometry analysis.Quantitative real-time PCR was used to evaluate the tran-scription levels of TLR1-10, IL-1R1, IL-1RAP, IL-6Rα, gp130, TNFR1, MyD88 and RIP1 in CD4+T cells.Plasma concentrations of IL-1β, IL-6 and TNF-αwere measured by enzyme-linked immunosorbent as-say.Results (1) The percentages of Treg cells and the levels of IL-10, TGF-β, CTLA4 and forkhead box P3 (Foxp3) in patients with acute KD were lower than those of healthy subjects (P<0.05), while the poly-ubiquitination of Foxp3 protein was significantly enhanced in patients with acute KD [(0.52 ±0.19) vs (0.08±0.02),P<0.05].Meanwhile, the four former items in KD patients with coronary artery lesions (KD-CAL+) were lower than those of KD patients without coronary artery lesions (KD-CAL-) (P<0.05), while the polyubiquitination level of Foxp3 protein in KD-CAL+group was much higher than that of KD-CAL-group [(0.70±0.28) vs (0.43±0.17), P<0.05].The levels of Treg cells, IL-10, TGF-βand CTLA4 in patients with KD were increased and the ubiquitination of Foxp3 protein was inhibited [(0.24±0.10) vs (0.52±0.19), P<0.05] upon the treatment with IVIG.(2) The levels of STUB1 and HSP70 in CD4+T cells were significantly elevated during acute KD, while the levels of USP7 were decreased (P<0.05).The ratios of STUB1/USP7 in patients with acute KD were much higher than those of the control group [(2.65± 0.92) vs (1.09±0.37), P<0.05], but were significantly decreased after IVIG therapy [(1.46±0.53) vs (2.65±0.92), P<0.05].A negative correlation was found between STUB1/USP7 ratio and Foxp3 level during acute KD (r=-0.56, P<0.05).Moreover, KD patients with CAL+showed higher levels of STUB1 and HSP70 and higher ratios of STUB1/USP7 (P<0.05), but lower levels of USP7 as compared with those of KD-CAL-group (P<0.05).(3) The plasma concentrations of inflammatory cytokines (IL-1β, IL-6 and TNF-α), the levels of surface receptors ( IL-1R1/IL-1RAP/TLR4, IL-6Rα/gp130 and TNFR1) and its downstream molecules ( MyD88, pSTAT3 and RIP1) in CD4+T cells were up-regulated during acute KD ( P<0.05), especially in patients with CAL+, but were down-regulated upon the IVIG therapy (P<0.05).No significant differences with other TLRs were found among the groups (P>0.05).Conclusion Hyper-ubiq-uitination of Foxp3 protein might be involved in the immune dysfunction during Kawasaki disease.