2.Relationship between Anemia,Iron Deficiency and Helicobacter Pylori Infection in Children
hai-feng, DONG ; xing-xian, YANG ; yan, DING ; yuan, QIN
Journal of Applied Clinical Pediatrics 2006;0(15):-
Objective To investigate the prevalence features of helicobacter pylori (Hp) infection,anemia and iron deficiency in a po-pulation of Wuhan children with 2 to 6 years old,and the relationship between Hp infection and anemia,iron deficiency in the children.Methods Randomly taking 95 children who had taken tests in our hospital's check-up centre in 2008 as the study objects,2 kinds of exa-mination were employed to detect Hp infection.Serum levels of Hp-IgG were measured by enzyme linked immunosorbent assay (ELISA) methods to evaluate past infection.The 14C urea breath test (14C-UBT) was conducted to obtain information of the presence of current/active Hp infection.In the morning 3 mL fasting venous blood was collected to determine the serum levels of Hp-IgG antibodies and ferritin.Hemoglobin values were determined with a hemoglobinometer.Serum levels of C-reactive protein (CRP) were tested in order to determine whether the children had evidence of current inflammation or infection.In addition,demographic information such as age and gender of the children and information about their use of antibiotics within the prior month were recorded.All cases were divided into 2 groups including the Hp infection group and non-Hp infection group according to laboratory examinations,then the Logistic regression was applied to analyze the relationship between Hp infection and anemia,iron deficiency.The Kappa identity test was taked to compare the 2 measures.Results Of the 95 children,18.9% were anemic and 36.8% were iron deficient.Forty percent of the cohort had Hp-IgG antibodies,74.4% tested positive by the UBT.Presence of Hp-IgG emerged as a significant risk factor for anemia,iron deficiency in adjusted analysis controlling for demographic factors,current inflammation,and antibiotic use.Conclusions Findings from different measure of Hp may reflect different stages of infection,with UBT results reflecting an earlier stage of infection,and presence of Hp-IgG reflecting established Hp infection associated with anemia,iron deficiency.
3.Effect of Paridis Rhizoma total saponins on apoptosis of human gastric cancer cell MKN-45 and Fas/FasL signaling pathway.
Hai-yan FANG ; Xiao-yan GONG ; Xing-hui HONG ; Ming-liu HUA ; Jin-ling HUANG
China Journal of Chinese Materia Medica 2015;40(7):1388-1391
OBJECTIVEThe study aimed to test if Paridis Rhizoma total saponins (PRTS) could induce apoptosis of human gastric cancer cell MKN-45.
METHODBased on the previous researches, PRTS was set by different concentrations to treat human gastric cancer cell for 12 h (5, 10, 20 mg x L(-1)). Fluorescent staining methods were adopted to observe apoptotic morphological changes of MKN-45. The apoptosis rates were analyzed by flow cytometry with Annexin V-FITC/PI staining. The enzymatic activities of caspase-3 and caspase-8 were measured by ELISA. The protein levels of Fas and FasL were detected by Western blotting.
RESULTUnder a fluorescence microscope, MKN-45 treated by PRTS was seen typical apoptotic morphological features. PRTS significantly increased the rate of apoptosis. Compared with the control group, there exsited significant differences in apoptosis rate of PRTS concentration of 20 mg x L(-1) (P < 0.01); besides, the enzymatic activities of caspase-3 and caspase-8 were promoted obviously after the effect of PRTS on MKN-45 cells for 12 h (P < 0.01). The protein levels of Fas and FasL in the MKN-45 were upgraded significantly.
CONCLUSIONPRTS can induce apoptosis of human gastric cancer cell MKN-45 , which is concerned with caspase-3 and caspase-8 and upgraded Fas and FasL.
Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Caspase 8 ; genetics ; metabolism ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Fas Ligand Protein ; metabolism ; Humans ; Magnoliopsida ; chemistry ; Rhizome ; chemistry ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Stomach Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; fas Receptor ; metabolism
4.Isolation of Acidithiobacillus ferrooxidans by Using Solid-plate Magnetophoresis
Xin-Xing LIU ; Wen-Bin LIU ; Ying YAN ; Hai-Yan WU ;
China Biotechnology 2006;0(03):-
Acidithiobacillus ferrooxidans are able to synthesize intra-cellular electron-dense magnetite,which makes possible that the bio-nano-magnetic particles could be obtained by cultivating Acidithiobacillus ferrooxidans. In order to isolate the strain which has the capacity to produce more magnetic particles, the solid-plate magnetophoresis method was firstly created. After isolation using the method, the rate of the cells which contain intra-cellular magnetic particles was increased from 30% to more than 90%, in addition, after isolation each cell possessed 2~5 magnetic particles which disperse in cells. The isolated cells are able to orient and migrate to the magnet in artificial magnetic field but could not orient swimming only under the geomagnetic field. Magnetosomes produced by Acidithiobacillus ferrooxidans were range from 40nm to 90 nm according to the results of TEM. Energy dispersive X-ray analysis indicated that extracted magnetic particles consisted of oxygen and iron. The results show that some Acidithiobacillus ferrooxidans cells have weak magnetotaxis and they could be able to be separated by solid-plate magnetophoresis method from others. With the development of this new isolation method, it is possible to do deeper research to generate a comprehensive description of the mechanism that how Acidithiobacillus ferrooxidans synthesize the magnetic particles.
5.Antibiotic resistance phenotypes and genotypes of ESBLs and AmpC ?-lactamases from Klebsiella oxytoca
Wen-Li ZHANG ; Ding-Xia SHEN ; Xing-Ping LI ; Yan-Ping LUO ; Jun LIU ; Xin-Yu WANG ; Xing-Hai SHI ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To study antibiotic resistance phenotypes and genotypes of extended spectrum ?-lactamases (ESBLs) and AmpC ?-lactamase-producing Klebsiella oxytoca isolated from specimens of respiratory tract in children.Methods Bacterial isolates were identified by API or VITEK32. Agar dilution was used for antibiotic susceptibility test,and ESBLs and AmpC were detected by confirmatory test recommended by CLSI/NCCLS and by 3-aminophenylboronic acid (APB) disk potentiation test, respectively.Microarray was used to determine the genotypes of ESBLs and AmpC ?-lactamases.Genotypes of Klebsiella oxytoca were determined by enterobacterial repetitive intergenic consensus (ERIC)- PCR.Results ESBLs were positive in 129 out of 165 isolates (78.2%).Both ESBLs and AmpC ?- lactamases were positive in 16 out of 165 isolates (9.7%).AmpC ?-lactamase alone producer was not detected in term of phenotype and genotype.CTX-M was the most common type of ESBLs and DHA was the only type of AmpC ?-lactamase in these isolates.Most antibiotic resistant strains of Klebsiella oxytoca possessed the same genotype by ERIC-PCR.Although all strains were susceptible to carbpenem,Klebsiella oxytoca with ?-lactamases were more resistant to other antibiotic agents than those without ?- lactamases.Conclusions There is high prevalence of ESBLs production among Klebsiella oxytoca isolated from children in Urumqi.The main genotypes of ESBLs and AmpC ?-lactamases are CTX-M and DHA.
6.Clinical characteristics and therapy of idiopathic hypoparathyroidism in 30 infants.
Hai-Hua YANG ; Yong-Xing CHEN ; Hai-Yan WEI
Chinese Journal of Contemporary Pediatrics 2015;17(10):1137-1139
Female
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Humans
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Hypoparathyroidism
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blood
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diagnosis
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drug therapy
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Infant
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Infant, Newborn
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Male
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Parathyroid Hormone
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blood
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Retrospective Studies
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Vitamin D
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therapeutic use
7.Preparation and evaluation of risperidone-loaded microsphere/sucrose acetate isobutyrate in situ forming complex depot with double diffusion barriers.
Xia LIN ; Xing TANG ; Yu-hong XU ; Yu ZHANG ; Yan ZHANG ; Hai-bing HE
Acta Pharmaceutica Sinica 2015;50(6):775-782
In the present study, a risperidone loaded microsphere/sucrose acetate isobutyrate (SAIB) in situ forming complex depot was designed to reduce the burst release of SAIB in situ forming depot and to continuously release risperidone for a long-term period without lagime. The model drug risperidone (Ris) was first encapsulated into microspheres and then the Ris-microspheres were embedded into SAIB depot to reduce the amount of dissolved drug in the depot. The effects of different types of microsphere matrix, including chitosan and poly(lactide-coglycolide) (PLGA), matrix/Ris ratios in microspheres and morphology of microspheres on the drug release behavior of complex depot were investigated. In comparison with the Ris-loaded SAIB depot (Ris-SAIB), the complex depot containing chitosan microspheres (in which chitosan/Ris = 1 : 1, w/w) (Ris-Cm-SAIB) decreased the burst release from 12.16% to 5.80%. However, increased drug release rate after 4 days was observed in Ris-Cm-SAIB, which was caused by the high penetration of the medium to Ris-Cm-SAIB due to the hydrophilie of chitosan. By encapsulation of risperidone in PLGA microspheres, most drugs can be prevented from dissolving in the depot and meanwhile the hydrophobic PLGA can reduce the media penetration effect on the depot. The complex depot containing PLGA microspheres (in which PLGA/ drug=4 : 2, w/w) (Ris-Pm-SAIB) showed a significant effectiveness on reducing the burst release both in vitro and in vivo whereby only 0.64% drug was released on the first day in vitro and a low AUC0-4d value [(105.2± 24.4) ng.mL-1.d] was detected over the first 4 days in vivo. In addition, drug release from Ris-Pm-SAIB can be modified by varying the morphology of microspheres. The porous PLGA microspheres could be prepared by adding medium chain triglyceride (MCT) in the organic phase which served as pore agents during the preparation of PLGA microspheres. The complex depot containing porous PLGA microspheres (which were prepared by co-encapsulation of 20% MCT) (Ris-PPm-SAIB) exhibited a slightly increased AUC0-4d of (194.6±15.8) ng.mL-1d and high plasma concentration levels from 4 to 78 days [Cs(4-78d)=(7.8±1.2) ng.mL-1]. The plasma concentration on 78 day C78d was (9.0 2.5) ng.mL-1 which was higher than that of Ris-Pm-SAIB [C78d= (1.6 ± 0.6) ng.mL-1]. In comparison with Ris-Pm-SAIB, the AUC4-78d of Ris-PPm-SAIB increased from (379.0±114.3) ng.mL-1.d to (465.0 ±149.2) ng.mL-1.d, indicating sufficient drug release from the Ris-PPm-SAIB. These results demonstrate that the risperidone loaded porous PLGA microsphere/SAIB in situ forming complex depot could not only efficiently reduce the burst release of SAIB depot both in vitro and in vivo, but also release the drug sufficiently in vivo, and be capable to continuously release the drug for 78 days.
Chitosan
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Drug Carriers
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Lactic Acid
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Microspheres
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Polyglycolic Acid
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Risperidone
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chemistry
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Sucrose
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analogs & derivatives
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Technology, Pharmaceutical
8.Matrix metallo proteinase-2 and focal adhesion kinase expression in herpes simplex virus-1 infected human corneal epithelial cell
Ting, CAO ; Yi-qiao, XING ; Yan-ning, YANG ; Hai-feng, MEI
Chinese Journal of Experimental Ophthalmology 2013;32(11):1050-1054
Background Previous studies showed that after herpes simplex virus-1 (HSV-1) infection of the cornea,matrix metallo proteinase-2 (MMP-2) (produced by corneal cells and corneal epithelial cells) plays an important role in the development of HSK.Focal adhesion kinase (FAK)plays an important role on the expression,release and activation of MMPs.This study explored the expressions of MMP-2 and FAK,which induced by HSV-1 infected human corneal epithelial cells.Objective To investigate MMP-2 and FAK expression in HSV-1 infected human corneal epithelial cells.Methods Human corneal epithelial cells were infected with HSV-1 in vitro to establish cell model of viral infection.The expression of MMP-2 and FAK were detected by reverse transcription PCR (RT-PCR),Western blot,immunohistochemical method and immunofluorescence method at 2 hours,20 hours and 40 hours after infection.Results At 2 hours,20 hours and 40 hours after infection,the expressionis of MMP-2 mRNA and FAK mRNA were significantly increased in comparison with uninfected cells (MMP-2 mRNA:Ftime =0.968,P=0.436 ; Fgroup =47.649,P =0.000 ; Fi ion =0.757,P =0.536.FAK mRNA:Ftime =0.658,P =0.631 ; Fgroup =35.182,P=0.000;Finteraction =1.386,P=0.137).Western blot assays showed that there were no significant differences in p-FAK,FAK or MMP-2 expressions between infected cells and control cells after 2 hours (P>0.05),but the expression levels of infected cells were significantly increased at 20 hours and 40 hours (both at P < 0.05).Immunohistochemistry results showed that longer infection time was associated with an increased number of cells staining for MMP-2,FAK and p-FAK.Conclusions At the initial stage of HSV-1 infected,p-FAK plays an important role in the process of virus invading and MMP-2 activation.
9.Expression of Truncated NSP2 Protein of Porcine Reproduction and Respiratory Syndrome Virus in E.coli and Preparation of Monoclonal Antibodies Against NSP2 Protein
Hai-Yan WANG ; Ping JIANG ; Yi-Jun DU ; Yu-Feng LI ; Jun-Xing LI ; Fang SHEN ;
China Biotechnology 2006;0(02):-
The NSP2 gene of porcine reproductive and respiratory syndrome virus (PRRSV)S1 strain was partly amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a fusion protein GST-tNSP2 with molecular weight of 50 kDa was expressed in E.coli. The purified GST-tNSP2 protein showed a strong reaction with the PRRSV-positive sera in Western blot assay. Balb/c mice were immunized with the purified protein, and the splenocytes of the immunized mice were fused with murine myeloma cells SP2/0. After subcloning by 3 times, two hybridoma clones which produced McAbs steadily were screened by ELISA, named 3H3 and 2B5. They all reacted strongly with the PRRSV S1 infected Marc-145 cells in IFA, but not with the PRRSV SY0608 strain. Both of the McAbs belong to IgG1 isotype, and their light chains belong ? type. The expressed GST-tNSP2 protein and McAbs could be used for identification of PRRSV isolates and functional analysis of NSP2.
10.Expression of NSP 3AB Gene of Encephalomyocarditis Virus(EMCV)in E.coli and Development of Monoclonal Antibodies Against 3AB Protein
Fang SHEN ; Ping JIANG ; Yu-Feng LI ; Jun-Xing LI ; Hai-Yan WANG ;
China Biotechnology 2006;0(09):-
Objective:To express the EMCV 3AB gene by prokaryotic expression systerm,and prepare monoclonal antibodies against it. Method: NSP 3AB gene of Encephalomyocarditis virus (EMCV) was amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a recombinant protein 3AB with high antigenicity was expressed in E.coli. Balb / c mice were immunized by purified recombinant 3AB protein of inclusion-body, and the splenocytes of the immunized mice were fused with murine myeloma cells to produce hybridoma cell line. Results: After subcloning by 3 times, one strain of hybridoma cell line steadily secreting antibodies of 3AB protein was obtained, named 2D12. The McAb belongs to IgG1/?. The McAb and was confirmed by indirect immunofluorescent assay (IFA) and Western blot. Conclusion: These results can provide a potential value for structural and functional studies of EMCV-3AB and early diagnosis of Encephalomyocarditis virus infection.