Objective To construct and express a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum(Sj) in Escherichia coli(E.coli) BL21 (DE3).Methods Total RNA was extracted from Sj adult worms by RNeasy Mini kit,26 kilodalton glutathione-S-transferases of Schistosomajaponicum (Sj26GST) antigen gene was amplified by real-time PCR(RT-PCR) from the total RNA,then cloned into a prokaryotic expression plasmid pGEX1λT and transformed into E.coli BL21 (DE3) to construct pGEX-Sj26GST; BL21 (pGEX-Sj26GST) was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG),and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and Western blotting.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and restriction enzyme double-digestion technique confirmed that Sj26GST antigen gene was successfully cloned into pGEX-1λT vector,the relative molecular mass of the expressed recombinant protein was approximately 52 × 103 by SDS-PAGE,and the amount of expressed protein was 20％ of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pGEX-Sj26GST is successfully constructed and highly expressed in E.coli and the expressed fusion protein shows specific antigenicity.

OBJECTIVETo explore the protective effect and mechanism of total flavones of Bidens pilosa L. (TFB) on IgA1 induced injury of venous endothelial cells in Henoch-Schönlein purpura (HSP) children patients. METHODS Human umbilical venous endothelial cells (HUVECs) were taken as subject. They were intervened by normal IgA1 and HSP children patients' serum IgA1, and added with different concentrations TFB at the same time. Then they were divided into the blank control group, the normal control group, the HSP IgA1 group, and HSP IgA1 plus TFB (1.0, 0.5, 0.25 mg/mL) groups. Levels of TNF-α and IL-8 in supernate were detected by ELISA. The NO level was detected by nitrate reductase method. mRNA and protein expressions of NF-κB and ICAM-1 in HUVECs were detected by fluorescent quantitative PCR and Western blot respectively.

RESULTSCompared with the normal control group and the blank control group, levels of IL-8, TNF-α, and NO all significantly increased in the HSP group (P < 0.05). Compared with the HSP group, levels of IL-8, TNF-α, and NO significantly decreased after intervention of TFB (1.0 and 0.5 mg/mL; P < 0.05, P < 0.01). Results of fluorescent quantitative PCR and Western blot showed, as compared with the blank control group and the normal control group, mRNA and protein expressions of NF-κB and ICAM-1 in HSP children patients' serum IgA1 induced venous endothelial cells significantly increased with statistical difference (P < 0.05, P < 0.01). Compared with the HSP group, mRNA and protein expressions of NF-KB and ICAM-1 were obviously down-regulated after intervention of TFB (1.0, 0.5, 0.25 mg/mL), with statistical difference (P < 0.05, P < 0.01).

CONCLUSIONTFB could protect vascular damage by inhibiting in vivo high expression of NF-κB, reducing the production of IL-8, TNF-α, and NO in vascular endothelial cells of HSP children patients.

Bidens ; chemistry ; Child ; Flavones ; pharmacology ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Immunoglobulin A ; blood ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-8 ; metabolism ; NF-kappa B ; metabolism ; Nitric Oxide ; metabolism ; Purpura, Schoenlein-Henoch ; blood ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

One hundred and seventy-two strains of endophytic fungi were isolated from Taxus mairei,Cephalotaxus fortunei and Torreya grandis cv.merrillia.The result of the antifungal assay shows that ninety strains of the fungi have antagonism against one or more botanical pathogenic fungi,such as Neurospora sp.,Trichoderma sp.,Fusarium sp.etc.The percentage of antifungal strains to tested strains are as follows:40% Cephalotaxus fortunei,54.2% Taxus mairei,57.1% Torreya grandis cv.merrillia.Thirty-five strains have high antifungal activities,and their inhibition zone diameter is at least 15mm.The active endophytic fungi were identified as 18 genera,most of which belong to Paecilomyces and Fusarium etc.

Objective To observe the metabolism and distribution of arsenic in liver and brain of offspring rata by exposure to arsenic of pregnant rats or lactation dams and weaned pups,and explore if arsenic could penetrate the placental barrier,lactation barrier and blood brain barrier. Methods The Wistar female rots were randomly divided into four groups according to body weights,12 in each group,and were fed with drinking water that contained arsenic(NaAsO_2) 0,10,50,100 mg/L beginning from the gestafional day 6 until pups 42 days old. Pups were separately sacrificed on postnatal day(PND) 0,15,28,42. Arsenic in liver and brain of offspring rots and in breast milk was examined by atomic absorption speetrophotometer with an arsenic speeiation pretreatment system. Results Concentration of iAs,MMA,DMA of brain in 50,100 mg/L groups were higher than that of 0 mg/L group[0,0,0,(7.3±6.6),0,(44.2±27.4)ng/g]on PND 0,42[iAs: (120.0±46.0),(195.5±125.3),(216.5±278.4),(176.6±151.8) ng/g; M MA: (47.2±18.1),(199.6±389.1),(47.4±55.2),(82.7±79.2) ng/g; DMA: (984.3±377.4),(2222.1±1433.2),(998.1±368.3),(1781.3±715.7)ng/g,all P < 0.05]. Concentration of DMA of brain in 50,100 mg/L groups were higher than that of 0 mg/L group[(13.9±18.1),(50.6±98.3)ng/g]on PND 15,28 [(270.3±73.1),(323.9±72.7),(758.7±245.9),(1020.6±383.6) ng/g,all P < 0.05]. Concentration of iAs,DMA of liver in 10,50,100 mg/L groups were higher than that of 0 mg/L group [(1.4±3.5),(49.7± 47.1),0,(100.4±30.2)ng/g]on PND 28,42 [iAs: (37.5±28.1),(268.8±246.4),(307.2±339.9),(15.4±9.4),(479.1±161.1),(408.4±51.9)ng/g;DMA: (594.5±148.8),(3181.9±519.0),(4834.2±2568.4),(1061.8± 85.2),(3697.1±553.7),(4120.0±732.8) ng/g,all P < 0.05]. Concentration of DMA of liver in 10,50,100 mg/L groups were higher than that of 0 mg/L group[(13.2±20.5)ng/g]on PND 15[(182.0±60,2),(637.6±90.0),(1458.7±196.3)ng/g,all P < 0.05]. Concentration of arsenicals of liver and brain showed a dose-dependent increase. The concentrations of DMA of breast milk in 50,100 mg/L groups were also higher than that of 0 mg/L group[(9.8±13.4),0 ng/g]on PND 0,15 [(182.3±85.9),(372.2±203.9),(124.2±33.1),(244.4±196.5)ng/g,all P < 0.05]. In the analysis of the change of arsenic on different postnatal day,we found the concentration of iAs,MMA,DMA,TMA in liver and brain of pups all decreased on postnatal day 15,and was lower than that on PND 0,28 and 42. Conclusions The distribution of arsenic and methyl-metabolism in liver and brain of pups is related with arsenic exposure dose. Arsenic can penetrate the placenta and blood brain barrier easily and lactation can hinder arsenic intake in some extent.

OBJECTIVETo study the oxidative stress induced by consumption of mercury-contaminated rice in rats, and to assess the possible public health risk of mercury contamination in Wanshan mining area.

METHODSSprague Dawley rats were fed the mercury-contaminated rice produced from Wanshan area for 90 days. The antioxidant status and the free radicals in rat serum were evaluated.

RESULTSHigh mercury accumulation in organs of rats fed the mercury-contaminated rice confirmed the server pollution of mercury in Wanshan mining area. The intensity of electron spin resonance (ESR) signal increased by 87.38% in rats fed the rice from Wanshan compared with that in the control rats fed the rice from Shanghai, suggesting that chronic dietary consumption of rice from mercury mining area could induce an aggravation of free radicals. Feeding the mercury-contaminated rice was associated with significant decreases in the antioxidant enzymatic activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and concentration of serum nitric oxide (NO), but it had no effect on serum nitric oxide synthase (NOS) activity. Feeding the mercury-contaminated rice raised the level of serum malonyldialdehyde (MDA), indicating the occurrence of oxidative stress.

CONCLUSIONThe long-term dietary consumption of mercury-contaminated rice induces the aggravation of free radicals and exerts oxidative stress.

Animals ; Brain ; metabolism ; China ; Environmental Pollutants ; analysis ; pharmacokinetics ; toxicity ; Food Contamination ; analysis ; Free Radicals ; blood ; Glutathione Peroxidase ; blood ; Industrial Waste ; adverse effects ; Kidney ; metabolism ; Liver ; metabolism ; Malondialdehyde ; blood ; Mercury ; analysis ; pharmacokinetics ; toxicity ; Methylmercury Compounds ; analysis ; pharmacokinetics ; toxicity ; Nitric Oxide ; blood ; Nitric Oxide Synthase ; blood ; Oryza ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood

OBJECTIVETo investigate the material base and underlying mechanism of the effect of cluster needling of scalp acupuncture on the neuronal plasticity in rats with focal cerebral infarction.

METHODSThe model rats with acute cerebral infarction were made by blocking the middle cerebral artery with monofilament. One hundred and thirty two Wistar rats were randomly divided into 4 groups: sham-operation group (A), model group (B), point-to-point scalp acupuncture group (C) and cluster-needling of scalp acupunture group (D). Puncturing from "Baihui (GV 20)" to "Qubin (GB 7)" was used in group C. Cluster needling of scalp acupuncture was used in group D, in which needles were inserted forward and slantingly into "Baihui (GV 20)" and its left and right sides at 4 mm. In both groups, the treatment was carried out with rapid twirling reinforcing-reducing for 1 min then retaining needle for 30 min, once a day, 6 days in one course, for treating 4 courses. There was no treatment for group A and B. The change of neurological function was evaluated with Bederson score, while the expression of microtubule-associated protein 2 (MAP-2) in the ischemic penumbra was examined with immunohistochemistry (streptavidin-peroxidase method).

RESULTSIn comparison,with group B, the score of neurological function in group D decreased on 7th day (P<0.05), while the scors in group C and D also decreased on 14th and 28th days (both P<0.05). As compared with group C, the score of neurological function in group D obviously decreased on 28th days (P<0. 05). Comparing with group B, the expression of MAP-2 on the ischemic cortex was significantly increased in group D and C on 7th, 14th and 28th days (all P<0. 05), however, this expression in group D was higher than that in group C on 14th and 28th days (P<0. 05).

CONCLUSIONCluster needling of scalp acupuncture can improve the neurological function of rats with focal cerebral infarction, and increase the expression of MAP-2 in the ischemic penumbra.

Acupuncture Therapy ; methods ; Animals ; Cerebral Infarction ; ethnology ; metabolism ; physiopathology ; therapy ; Disease Models, Animal ; Male ; Microtubule-Associated Proteins ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Scalp

AIMTo establish a comprehensive HPLC analytical method of Huanglianjiedu decoction.

METHODSThis study was performed by HPLC-UV/MS to identify the chemical constituents of the whole and individual herbs of the "Huanglianjiedu decoction". Zorbax Extend C18 (150 mm x 4. 6 mm ID, 5 microm) column was used; the mobile phase was composed of acetonitrile (A) and water (B, with 0.5% acetic acid) with gradient elution; the flow rate was 1.0 mL x min(-1) and the column temperature was setup at 25 degrees C. The detection wavelength was 254 nm.

RESULTSThe chromatogram of Huanglianjiedu decoction showed 21 main peaks. Peaks 1, 2, 5 and 18 were from Gardenia jasminoides Ellis, Peaks 8, 13, 14, 15, 16, 17, 19 and 21 from Scutellaria baicalensis Georgi. While 10 from Coptis chinensis Franch and 20 from Phellodendron amurense Rupr., Peaks 3, 4, 6, 9, 11 and 12 came from them together. Peak 7 presented in the chromatograms of the herbs except Gardenia jasminoides Ellis. By comparison of the retention time, the on-line UV spectra and MS spectra, 11 peaks were identified as 5 (geniposide), 9 (jatrorrhizine), 10 (coptisine), 11 (palmatine), 12 (berberine), 13 (baicalin), 15 (oroxin A), 17 (wogonoside), 19 (baicalein), 20 (obaculactone), 21 (wogonin), then eight of them were quantified by HPLC-UV.

CONCLUSIONThe method could represent the characteristics of Huanglianjiedu decoction, and it could be used to evaluate the quality and quantity of Huanglianjiedu decoction. It distinguished between Coptis chinensis Franch and Phellodendron amurense Rupr. by HPLC for the first time.

Berberine ; analogs & derivatives ; analysis ; Berberine Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; methods ; Coptis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Gardenia ; chemistry ; Mass Spectrometry ; methods ; Phellodendron ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Scutellaria baicalensis ; chemistry ; Spectrophotometry, Ultraviolet ; methods

OBJECTIVETo investigate the main components of inner ear antigens inducing autoimmune Meniere's disease (AIMD) in guinea pigs.

METHODSThe guinea pigs were immunized with isologous crude inner ear antigens (ICIEAg). Then, the hearing function was measured with auditory brainstem response (ABR), the vestibular function was measured with electronystagmography (including spontaneous nystagmus and caloric test), and inner ear histopathological changes were observed by inner ear celloidin section with haematoxylin-eosin staining and observed under light microscope. According to these results, the AIMD-model animals from non-AIMD-model ones were distinguished. The special antibodies against ICIEAg in sera were measured with ELISA. The antigen-antibody reactions against different components of ICIEAg were detected by Western blotting with sera of AIMD and non-AIMD guinea pigs respectively. Then, we analysed the contrast between them and found the main components of the ICIEAg that were positive reaction in AIMD guinea pigs and negative reaction in non-AIMD guinea pigs.

RESULTSThe result of ELISA demonstrated that the sera of both the AIMD and non-AIMD guniea pigs contained the special antibodies against ICIEAg after immunized with ICIEAg. The difference of the amount of antibody against ICIEAg between AIMD guinea pig group and non-AIMD guinea pig group was not significant. Western blotting assay showed only the sera of AIMD guinea pig contained the antibodies against the specific antigens with the molecular of 68 000, 58 000, 42 000 and 28 000.

CONCLUSIONSICIEAg contain many different components, the AIMD might only happen in the guinea pigs in which the special immunization against the main components that could induce this kind of disorder appeared. The inner ear antigens with molecular of 68 000, 58 000, 42 000 and 28 000 might be the main components inducing AIMD in guinea pigs.

Animals ; Autoantigens ; immunology ; Autoimmune Diseases ; immunology ; Disease Models, Animal ; Ear, Inner ; immunology ; Guinea Pigs ; Labyrinth Diseases ; immunology

OBJECTIVETo determine actinoside C in the leaves of Actinidia kolomikta with different growth periods.

METHODThe separation was performed at 25 degrees C on ZORBAX Extend C18 column (4.6 mm x 250 mm, 5 microm), using amixture of methanol and water (51:49) as a mobile phase. The flow rate was 1.2 mL x min(-1), and the wavelength for measurement was 267 nm.

RESULTThe results showed that the contents of actinoside C in the leaves of A. kolomikta were variety in different growth periods. Actinoside C could reach its highest content in the middle ten days of June, then the content would decrease in the middle ten days of July slightly, it could reach their lowest content in the middle ten days of August.

CONCLUSIONThe optimal collective date for A. kolomikta are in the middle ten days of June.

Actinidia ; chemistry ; growth & development ; Chromatography, High Pressure Liquid ; methods ; Flavones ; analysis ; chemistry ; Glycosides ; analysis ; chemistry ; Molecular Structure ; Plant Leaves ; chemistry ; growth & development ; Plants, Medicinal ; chemistry ; growth & development ; Seasons

At present, the objective of cutting and pruning Cistanche deserticola is to harvest in successive years and enhance the harvesting yield and quality of C. deserticola in the process of the artificial cultivating C. deserticola. An experiment was conducted focusing on cutting and pruning C. deserticola in artificial forests of Haloxylon ammodendron drip-irrigated with saline water at the hinter-land of the Taklimakan desert, according to different growth stages and lengths. The results were following: (1) The effect of cutting on C. deserticola was similar to that of pruning, which resulted in three kinds of morphological types, not related to the bloom and size of C. deserticola. (2) The growth forms were diversified after pruning. Among them, there had sprouting new body, died or maintaining life with no sprouting, mildewed on its surface layer, etc. However, some of new bodies were sprouting from the lower part of the old body. The death rate of bloomed C. deserticola was higher than that of the underground, and the death rate of the 40 cm in stubble height for C. deserticola was higher than those with the stubble height of 20 cm and 5 cm. (3) Most of the diameter of living C. deserticola after pruning was increasing, but some of them changed little. (4) The mildew and rot of C. deserticola and the broken of the roots of the H. ammodendron and the fallen of the point of the inoculated when it was dug, which would cause the death of the C. deserticola. On the other, the yield-increasing effect and the economic benefit of the techniques of the pruning of Cistanche would need further research and evaluate. Therefore, the application of this technique needs to be cautious.
Amaranthaceae ; growth & development ; Cistanche ; growth & development ; Forests ; Fruit ; growth & development ; Plant Roots ; growth & development