2.Serial recombinant expression and activity against tumor cells in vitro of antibacterial peptide Alloferon-1.
Qi SUN ; Ai-hua SUN ; Jie YAN
Journal of Zhejiang University. Medical sciences 2008;37(1):60-66
OBJECTIVETo construct a prokaryotic expression system to serially express Alloferon-1 and to determine the anti-tumor activity of its products in vitro.
METHODSAn artificial fusion gene containing 6 x His-EK-8 x Alloferon-1-EK-6 x His sequences was constructed by linking primer PCR. By using routine molecular biological methods, the artificial fusion gene was cloned and its prokaryotic expression system was then constructed. SDS-PAGE and BioRad Agarose Image Analysor was applied to measure the expression and output of the target recombinant products 8 x rAlloferon-1-EK. Ni-NTA affinity chromatography and EK digestion and Sephadex G-50 chromatography were performed to extract 8 x rAlloferon-1-EK and rAlloferon-1-EK, respectively. The proliferation of KB, SGC and HL-60 tumor cells was tested by using MTT method after treatment with directly synthesized Alloferon-1 (sAlloferon-1), Aloferon-1-EK (sAlloferon-1-EK) and rAlloferon-1-EK.
RESULTThe target artificial fusion gene and its prokaryotic expression system pET42a-8 x rAlloferon-1-EK-E. coliBLDE3 with the expected sequences were obtained. Under inducement of IPTG, the prokaryotic expression system expressed the target serial recombinant protein 8 x rAlloferon-1-EK and its output was approximate 30 % of the total bacterial proteins. 8 x rAlloferon-1-EK and rAlloferon-1-EK were obtained through Ni-NTA and Sephadex G-50 columns. sAlloferon-1, sAlloferon-1-EK and rAlloferon-1ìrAlloferon-EK showed similar remarkable effects of inhibiting the growth and proliferation of KB, SGC and HL-60 cells in vitro within 25 approximately 100 microg/ml concentration range (P<0.01), and there were no significant differences in the inhibiting effects among the three agents (P>0.05).
CONCLUSIONA prokaryotic expression system to serially express rAlloferon-1 has been successfully constructed. The product rAlloferon-1-EK has a similar anti-tumor activity compared to both the synthesized Alloferon-1 and Alloferon-1-EK in vitro.
Animals ; Anti-Bacterial Agents ; pharmacology ; Antineoplastic Agents ; metabolism ; pharmacology ; Bacterial Vaccines ; administration & dosage ; genetics ; metabolism ; Cell Line, Tumor ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Growth Inhibitors ; genetics ; metabolism ; pharmacology ; HL-60 Cells ; Helicobacter pylori ; genetics ; metabolism ; Humans ; KB Cells ; Peptides ; genetics ; immunology ; metabolism ; pharmacology ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology
3.The effect of fluoride and arsenic pollution on bone metabolism in exposed population
Qi-bing, ZENG ; Yun, LIU ; Ai-hua, ZHANG ; Feng, HONG ; Jun NG YA ; Xian, YU
Chinese Journal of Endemiology 2011;30(4):393-395
Objective To explore the effect of fluoride and arsenic pollution on bone metabolism in exposed population. Methods One hundred and fifty-two fluoride and arsenic exposed people were selected from Jiaole village, Yuzhang town, Xingron county, Guizhou province in 2006, and 59 not exposed people from Daguoduo village 13 km away from Jiaole village were selected as control. Urinary fluorine(UF), urinary arsenic (UAs), urinary hydroxyproline (UHYP), cross-linked N-telopeptides of type I collagen (UNTX) and bone strength index(STI) were detected. Results The main effect of fluoride on UHYP and UNTX were statistically significant (F = 9.785, 4.225, P < 0.01 ), but was not significant on STI(F = 0.183, P > 0.05). The main effect of arsenic on UNTX was statistically significant (F = 2.660, P < 0.05 ), but was not significant on UHYP and STI(F = 2.012, 0.183,all P > 0.05). The interaction between fluoride and arsenic on UNTX was statistically significant (F= 2.429, P <0.01), but was not significant on UHYP and STI(F= 1.218, 1.001, all P> 0.05). Conclusions Fluoride exposure can affect the metabolism of collagen and bone resorption, and Arsenic exposure main affect bone resorption, fluoride and arsenic co-exposure have more significant effect on bone resorption. UNTX may be used as biological biomarker of bone metabolism for population co-exposed to fluoride and arsenic in health monitoring.
4.Comprehensive nursing interventions during perioperation period of renal biopsy in children
Chinese Journal of Modern Nursing 2009;15(33):3467-3470
Objective To explore the impact of nursing interventions during perioperation period of renal biopsy in children. Methods 120 patients were divided to intervention group and control group. Patients of intervention group received comprehensive interventions including health education, touch, psychological intervention and behavior intervention while those of control group received usual nursing intervention.Psychological state before operation, puncture success rate, co-operation level, and complications after operation were observed and compared between two groups. Results Compared with the control group, the intervention group had fewer psychological problems and fewer complications after operation while the puncture success rate and co-operation level were higher than those in the control group. Co-operation level of patients was significantly related to age but was not significantly related to gender. Conclusions Effective comprehensive nursing intervention during perioperation period of renal biopsy in childrencan improve the puncture success rate and decrease accident of complications after operation.
5.A study on the prevalence of the urinary incontinence among female adults in Hangzhou and its effect on life quality
Guang-Hua LAI ; Xiao-Long HE ; Xiao ZHANG ; Ai-Fen WANG ; Qi-Qi YU
Journal of Preventive Medicine 2016;28(4):328-331
Objective Toinvestigatetheprevalenceofurinaryincontinenceanditseffectonlifequalityamongfemale adultsinHangzhou,andtoprovideevidencetothecontroloffemaleurinaryincontinence.Methods Aquestionnaire survey was performed in Gongshu and Xiacheng districts in Hangzhou from October 2013 to June 2014,and 4 563 women aged over 20 years were interviewed through a questionnaire including International Consultation on Incontinence Questionnaire Lower Urinary Tract,IQ-FLUTS and demographic information.Single and multi factor logistic regression analysiswereusedtoanalyzetheriskfactorsofurinaryincontinence.Results Atotalof4785questionnairesweresent, and 4 563 effective questionnaires were recovered,with a recovery rate of 95.4%.The prevalence of urinary incontinence was 33.5%(1 530/4 563),and female with older age tended to have a higher prevalence of UI(P<0.01).Among which stress urinary incontinence (SUI ),urge incontinence (UUI )and mixed urinary incontinence (MUI ) were accounted for 20.2%(922/4 563),3.0%(135/4 563)and 10.3%(473/4 563)respectively.UUI and MUI had a greater effect on quality of life.According to the multi factor logistic regression analysis,gravidity,age,constipation,pelvic surgery and fat (BMI>24)wereriskfactorsforurinaryincontinence(P<0.05).Conclusion Becauseofthehighprevalenceofurinary incontinence among female adults,more attention should be paid to urinary incontinence suffers in order to improve the quality of life of female adults.
6.The change of NOS in pulmonary oxygen toxicity induced by different oxygen pressure.
Ai-Zi LIU ; Xiao-Chen BAO ; Yi-Qun FANG ; Zhong-Na SANG ; Hua-Jiang LI ; Wan-Qi ZHANG
Chinese Journal of Applied Physiology 2014;30(3):227-229
OBJECTIVELong time exhaled oxygen will induced oxygen toxicity. Some studies had found that different pathology may exised in normobaric and hyperbaric pulmonary oxygen toxicity, and nitric oxide synthase (NOS) may play a role. In this study, we discussed the change of NOS in normobaric and hyperbaric pulmonary oxygen toxicity.
METHODSSixty male SD rats were randomly divided into 6 groups (n = 10), exposed to 1 ATA (atmosphere absolute), 1.5 ATA, 2 ATA, 2.5 ATA and 3 ATA, 100% oxygen for 56, 20, 10, 8, 6 hours respectively. Rats were exposed to air as control. After exposure, the protein in bronchoalveolar lavage fluid (BALF), the wet/dry weight of lung and the expression of eNOS, nNOS in lung were defined.
RESULTSAs compared to air group, the protein in BALF, the wet/dry of lung were significantly elevated in 1.0 ATA group, while these changes were not so obviously in the other groups, and these changes in hyperbaric oxygen group (approximately 1.0 ATA) were significantly decreased as compared with nonnrmobaric oxygen group (1.0 ATA). The expression of nNOS were not changed in normobaric and hyperbaric pulmonary oxygen toxicity, while the expression of eNOS was significantly decreased in 2 ATA group, and significantly elevated in 2.5 ATA and 3 ATA group.
CONCLUSIONThe expression of eNOS can change when exposed to different pressures of oxygen.
Animals ; Disease Models, Animal ; Lung ; metabolism ; Male ; Nitric Oxide Synthase Type I ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Oxygen ; poisoning ; Pressure ; Rats ; Rats, Sprague-Dawley
8.Effect of combination therapy of methylprednisolone and cyclophosphamide on heart function and nursing in patients with refractory nephritic syndrome
Ai-Hua QI ; Hong ZHANG ; Rong CAO
Chinese Journal of Modern Nursing 2008;14(33):3459-3462
0bjective To evaluate the heart function of patients with refractory nephmtic syndrome during a combination therapy of methylprednisolone(MP)and cyclophosphamide(CTX).Methods From January 2004 to December 2007.30 patients with refractory nephrotic syndrome.who were in hospital and received a combination therapy of MP and CTX.were recorded for their heart rate and blood pressure before and during treatment,as well as observed for the complications.ResulIs Comparing with those before treatment.patients' heart rates were much quickly and their blood pressure were remarkablv higher during the combination treatment. And there was a statistic siglnificance(P<0.05).When patients received MP treatment.complications.such as cardiopalmus and dizziness,proned to emerging in the third cycle.When they received CTX treatment, complications,just like sicchasia,vomit and cardiopalmus.appeared in the first cycle.In addition. Myelosuppression and lesion of liver function often occurred during the sequence therapy.Conclusions When patients received a combination therapy of MP and CTX,we should monitor their heart funcion SO as to promptly make propotional measure and prevent complications according to variation of heart rate and blood pressure.
9.Effect of insulin-like growth factor-1 on hyperoxia-induced apoptosis in A549 cells.
Zhen-Ai JIN ; Yi-Kun ZHAO ; Zheng-Yong JIN ; Hai-Feng QI ; Mokhles Wali GUL ; Ming-Hua GONG
Chinese Journal of Contemporary Pediatrics 2013;15(6):490-493
OBJECTIVETo investigate the effect of insulin-like growth factor-1 (IGF-1), which can promote cell differentiation and inhibit cell apoptosis, on hyperoxia-induced apoptosis in A549 cells and its anti-apoptotic mechanism.
METHODSA549 cells were sub-cultured, exposed to hyperoxic conditions and were then treated with different concentrations of IGF-1 (1, 10, and 100 ng/mL) for 48 hours. Cell viability was measured by MTT assay. Cell apoptosis was evaluated by Annexin V-FITC/PI double-staining flow cytometry. Expression levels of Bax and Bcl-2 were measured by flow cytometry.
RESULTSThe middle-dose and high-dose IGF-1 intervention groups had higher cell viabilities than the hyperoxic exposure group [(64±3)% and (88±4)% vs (51±3)%; P<0.05]. Compared with the air control group, the hyperoxic exposure group had a significantly higher apoptotic rate [(38.3±5.4)% vs (2.4±0.9)%; P<0.05], a significantly lower expression level of Bcl-2 [(72±5)% vs (91±4)%; P<0.05], and a significantly higher expression level of Bax [(54±6)% vs (3±2)%; P<0.05]. Compared with the hyperoxic exposure group, the low-dose, middle-dose, and high-dose IGF-1 intervention groups had significantly lower apoptotic rates [(16.1±4.7)%, (9.2±2.8)%, and (6.9±2.5)% vs (38.3±5.4)%; P<0.05], significantly higher expression level of Bcl-2 [(79±4)%, (94±4)%, and (100±5)% vs (72±5)%; P<0.05], and significantly lower expression level of Bax [(26±4)%, (5±2)%, and (4±2)% vs (54±6)%; P<0.05].
CONCLUSIONSHyperoxia significantly inhibits proliferation and promotes apoptosis in A549 cells. IGF-1 may promote cell proliferation and inhibit hyperoxia-induced apoptosis in A549 cells by regulating the expression of Bcl-2 and Bax.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Hyperoxia ; pathology ; Insulin-Like Growth Factor I ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; bcl-2-Associated X Protein ; analysis
10.Simultaneous determination of fumonisins B1 and B2 in traditional Chinese medicines by high-performance liquid chromatography-tandem mass spectrometry.
Ting-Ting XIE ; Feng QIU ; Mei-Hua YANG ; Ai-Di QI
Acta Pharmaceutica Sinica 2011;46(7):822-827
A rapid and sensitive analytical method was developed for the simultaneous determination of fumonisins B1 and B2 in traditional Chinese medicines by HPLC-MS/MS. The detection limits for fumonisins B1 and B2 were 0.25 ng x mL(-1) corresponding to 2 microg x kg(-1) in samples. Recoveries from different samples spiked with fumonisins B1 and B2 at levels ranging from 0.2 to 3 mg x kg(-1) were 84.0%-96.1% and 86.3%-99.3%, respectively. Among the total of 34 samples purchased from local markets, ten samples of which were visibly moldy samples due to inappropriate storage, and 24 were normal samples. The results showed that 6 of the visibly moldy samples and 5 of the normal samples were contaminated with total fumonisins at levels ranging 82.4-2349 microg x kg(-1) and 102-729 microg x kg(-1), respectively.
Chromatography, High Pressure Liquid
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Drug Contamination
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Drugs, Chinese Herbal
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chemistry
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Fumonisins
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analysis
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Medicine, Chinese Traditional
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Plants, Medicinal
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chemistry
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Tandem Mass Spectrometry