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Detection of serum specific IgE to crab allergen in different group people with extracted Eriocheir sinensis allergens

Haibo HUANG ; Weiwei CHEN ; Wei CHEN ; Juan LI ; Xuedi LI ; Qunying LIU

Chinese Journal of Biochemical Pharmaceutics.2010;31(2):128-130.

Purpose To detect serum specific IgE to crab allergen in different group people with extracted Eriocheir sinensis allergic proteins and to provide laboratorial evaluation for diagnosis and treatment of crab anaphylaxis.Methods Micro titer plates were coated with the allergic proteins extracted from crab.A total of 1907 serum samples from 3 group people were detected for specific IgE to crab allergens with indirect ELISA.The serological IgE level of different group people allergic to crab food wag compared and analyzed.Results 203 individuals were serum specific IgE positive in the detected 1 907 serum samples,and the positire rates Wag 10.65 percent(203/1 907).The statistical analysis showed that the difference of serum IgE positire rates among the 3 group people wag very significant(P＜0.01).Conclusion The detection of serum specific IgE using allergic proteins extracted from crab has diagnostic meaning,since it can be used as laboratorial evaluation for clinical diagnosis and treatment of crab anaphylaxis.

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Determination of trifluoroacetic acid in bivalirudin by HPLC

Xiaoming YANG ; Dongmei ZHAO ; Qiujiang DU ; Yuru LI

Chinese Journal of Biochemical Pharmaceutics.2010;31(2):126-127.

Purpose To establish a HPLC method for determination of the content of trifluoroacetic acid in bivalirudin.Methods Kromasil 100-5 C_(18) column(4.6 mm×200 mm,5 μm)and 0.07% phosphoric acid solution(pH 3.0)-methanol(98:2)as the mobile phase at a flow rate of 1.0 mL/min were used with a column temperature of 30 ℃,a detection wavelength of 210 nm and a injection volume of 20 μL.Results The standard curve of trifluomacetic acid was linear in the range of 198.4-992.0 μg/mL,r=0.999 9.The detection limit Was 40 ng.The average recovery rate of method was 99.9%,and RSD was 0.41%.Conclusion The method of content determination was practicable and accurate.It can be used for content determination of trifluoroacetic acid in bivalirudin.

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Modulation of CXCR4,and MCF-7 cells invasion potential in vitro by norcantharidin

Xiaofeng XU ; Wengping CHEN ; Ling XU ; Yiyan DING ; Yi WANG

Chinese Journal of Biochemical Pharmaceutics.2010;31(2):122-125.

Purpose To investigate the effect and mechanism of norcantharidin(NCTD)on invasion of human breast cancer cell line MCF-7 in vitro.Methods MTT assay Was used to determine MCF-7 cell proliferation,Transwell kit was used to determine cell migration,treated with NCTD on 200,400,600 μmol/L,to detect the expression of CXCR4 and CXCL12 in the control and treatment groups by real time PCR and Western blot.Results NCTD had inhibitive effects on proliferation and invasion of human breast cancer cell line MCF-7 in a dose-dependent manner,with the IC_(50) value of 582 μmol/L at 24 h,and the expression of CXCR4 Was deCreased in NCTD groups in vitro.Conclusion NCTD in vitro inhibits invasion and metastasis of human beast cancer cell line MCF-7 throush the down regulation to the expression of CXCR4.

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Protective effect of dl-praeruptorin A on focal cerebral ischemia in mice

Wushuang YANG ; Bogang TENG ; Lichao YANG ; Yu ZHOU ; Yao WANG ; Xin JIN

Chinese Journal of Biochemical Pharmaceutics.2010;31(2):118-121.

purpose To investigate the protective effect and character of dl-praeruptorin A(Pd-Ia)on focal cerebral ischemia in mice.Methods Transient focal cerebral ischemia in mice WaS induced by middle cerebral artery occlusion for 1.5 h.Pd-Ia was administered intraperitoneally either with multiple doses(1,5 and 10ms/ks)at 0.5 h before ischemia or single dose(5 ms/kg)at 0.5 h and 1 h before ischemic,the same time of ischemia,the same time of reperfusion,or 0.5 h and 1 h after reperfusion respectively.Neurological deficit score,infarct volume,brain edema,the activities of SOD and the contents of MDA were determined.Results Pretreatment with multiple doses(5 and 10 ms/ks)of Pd-Ia at 0.5 h before ischemia or single dose(5 mg/kg)of Pd-Ia at 0.5 h before ischemia,at the same time of ischemic,at the same time of reperfusion and 0.5 h after reperfusion significantly attenuated neurological deficit score,decreased infarct volume and alleviated brain edema,and the treatment at the time of reperfusion had the most marked effect.Pd-Ia(5 or 10 ms/ks)can significantly enhance the activities of SOD and lower the contents MDA.Conclusion dl-praeruptorin A has a neuroprotective effect on the injury in the acute phase of transient focal cerebral ischemia in mice,with optimal doses of 5 ms/ks and the optimal therapeutic time point of the same time of reperfusion.

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Effect of paeonol on inflammatory factors after focal cerebral ischemia-reperfusion injury in rats

Qing YANG ; Jibiao WU ; Xiumei ZHANG

Chinese Journal of Biochemical Pharmaceutics.2010;31(2):111-113.

Purpose To observe the effect of paeonol on inflammatory factors after focal cerebral ischemiareperfusion injury in rats and its possible mechanism of paeonol.Methods Focal cerebral ischemic model was established by MCAO in rats.After 2 hours of cerebral ischemia and 22 hours of reperfusion,the expression of TNF-α and IL-1β in brain was determined by immunohistochemistry.The content of TNF-α and IL-1β in serum was measured by ELISA.Results Compared with model group,the levels of TNF-α and IL-1β in brain and serum in rats in the paeonol groups were obviously decreased.Conclusion Paeonol could relieve brain damage induced by focal cerebral ischemia-reperfusion,which may be related to inhibiting the production of inflammatory factors.

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Study on gatifloxacin in situ pH-sensitive gel release in vitro

Yajing XU ; Rongfeng HU ; Hui YIN ; Yu GAO

Chinese Journal of Biochemical Pharmaceutics.2010;31(2):107-110.

purpose To investigate the release properties of gatifloxacin in situ pH-sensitive gel in vitro.Methods The improved paddle method and the membraneless model were applied in assessing the drug release behavior.Results The gel erosion and drug release were increased with the increase of surface area and shaking frequency.The cumulative quantities of gel erosion were well correlated with the cumulative release of drug loaded in the gel.Conclusion Gatifloxacin was released from in situ pH-sensitive gel with zero-order kinetics characters,and drug release was mainly controlled by gel erosion.

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Effect of Radix Rehmanniae Praeparata on footpad inflammation induced by complete Freund's adjuvant in Wistar rats

Xueyan WANG ; Xuesen WEN ; Di ZHANG ; Xiaoni KONG ; Min SHI

Chinese Journal of Biochemical Pharmaceutics.2010;31(2):103-106.

Purpose To investigate the anti-inflammatory effect of Radix Rehmanniae Praeparata(RRP)on the footpad inflammation induced by complete Freund's adjuvant(CFA)in rats.Methods CFA 100 μL were injected subcutaneously to the Wistar rats at the pad of right hindfoots.19 days later,the rats were daily siven RRP water extract(0.625,1.250,2.500 g/kg·bw)or dexamethasone(0.5mg/ks·bw)intragastrically.The changes of body weight and foot volume were measured.The indexes of organ and blood were determined at the 29th day,the foot pad was removed,and routine paraffin section was performed.Results The model rats kept foot swelling and lymphocyte infiltrating,and the platelet number decreased.The other indexes were statistically insignificant when compared to the controls.RRP did not display any anti-inflammatory effect on the swollon foots,but thoracic gland and spleen indexes were rescued,and platelet number and creatinine content were increased by RRP administration in a dose-dependent manner.The anti-inflammation of dexamethasone was conspicuous,but the side effects were also significant.Conclusion RRP may be plays an adjunctive action in herbal recipes to treat rheumatoid arthritis.

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Comparative study on bio-activity of polysaccharide from Pinus massoniana pollen and its sulfated derivative

Yue GENG ; Hui LIU ; Lihua SHI ; Zhihe LIU

Chinese Journal of Biochemical Pharmaceutics.2010;31(2):98-102.

Purpose To study the polysaccharide from Pinus massoniana pollen(PPM)and to compare its anti-tumor,immune modulation activities and scavenging qualities of free radical with its sulfated derivative(S-PPM).Methods PPM Wag chemically modified by chlorosulfonic acid-pyfidine method and their bioactivities were compared.Results The substituting degree of S-PPM was 1.47.Results showed that S-PPM was more powerful in inhibiting the growth of tumor cell in vivo and in vitro and in promoting T,B lymphocytes than PPM.But there Wag no remarkable difference in promoting phagocytosis of macmphages.S-PPM was stronger in scavenging superoxide anion radical than PPM but it wag vice versa in scavenging hydroxyl radical.Conclusion S-PPM inhibited the cancer cell growth mainly through specific immunity.Sulfate of PPM influenced its quality of scavenging free radicals greatly.

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Expression and activities analysis of a fusion protein CREKA/tTF

Yi SU ; Jianghua YAN ; Shengyu WANG ; Jie HE ; Min YE

Chinese Journal of Biochemical Pharmaceutics.2010;31(2):94-97.

Purpose To prepare a novel fusion protein of CREKA and tTF as a universal carrier targeting to cancer,and to analyze its activities.Methods CREKA and tTF gene were acquired by PCR,and inserted into plasmid pET22b(+)to construct recombinant plasmid CREKA/tTF/pET22b(+),and the fusion gene was expressed in E.coli BL21.The fusion protein Wag purified through Nickel-affinity chromatography column.After purifying,the fusion protein was refold by subsequent dialysis.The activities of the fusion proteins were measured by coagulation timing and quantitative fluorescence test in vitro.Results The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was obtained.The fusion protein was highly expressed in E.coli BL21.The coagulation of the fusion protein Was determined by the coagulation test.And the capability of the fusion protein effectively binding to clotted plasma proteins is identified in quantitative fluorescence test.Conclusion The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was built.The fusion protein CREKA/tTF with both TF and CREKA activity was successfully obtained.

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Preparation and bioactivity assay of mIL-4-SA fusion protein

Zhen ZHANG ; Pingping FA ; Jinlong LI ; Zhiming HU ; Jimin GAO

Chinese Journal of Biochemical Pharmaceutics.2010;31(2):90-93.

Purpose To prepare streptavidin-tagged mouse interleukin-4(mIL-4-SA)bifunctional fusion protein and to study on its bioactivity.Methods The mIL-4 gene was cloned by RT-PCR and cloned into pET21 vector to get mIL-4-SA-pET21 expression plasmid.The mIL-4-SA fusion protein was expressed in BL21 (DE3)host bacteria and purified through the Ni-NTA affinity chromatography and refolded by dilution and dialysis.The effect of mIL-4-SA fusion protein on mouse thymocytes proliferation was evaluated by MTY.Flow cytometric analysis was performed to detect the mIL-4-SA fusion protein on the biotinylated B16F10 tumor cells.Results The mIL-4-SA-pET21 vector was successful by constructed and the mIL-4-SA fusion protein was expressed in BL21(DE3)at about 35%of total bacterial proteins.The purity of mIL-4-SA Was about 95% through Ni-NTA.The mIL-4-SA fusion protein exhibited bifunctional activities,i.e.,stimulative effect for mouse thymocyte proliferation and SA-mediated high-affinity binding to biotinylated cell surfaces(anchoring modified rate Was about 96.69%).Conclusion The mIL-4-SA fusion protein was expected to be developed for the treatment of tumors.

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