3.Expression and purification of high purity soluble chimeric protein VEGI~+
Li-Li DING ; Rui-Li WEI ; Ji-Ping CAI ; You LI ;
Academic Journal of Second Military Medical University 1999;0(12):-
Objective:To prepare a novel vascular endothelial growth inhibitor-soluble chimeric protein VEGI~+,so as to lay a basis for studying its biological activity.Methods:Chimeric molecule VEGI~+ was constructed by grafting oligopeptide CTTH- WGFTLC to extraeellular region of VEGI(VEGI_(23-174)).Before ligation into pET30a(+)expression vector,PCR product of the recombinant gene was cloned into pGEM-T vector and verified by restriction enzyme digestion and DNA sequencing,then pET30a-VEGI was used to transfect BL21(modified E.coli strain).The chimeric protein was purified by metal affinity chro- matography.Western blotting and coomassie blue staining were used for protein identification.Results:The chimeric molecule VEGI~+ was confirmed by restriction enzyme digestion and DNA sequencing.The constructed pET30a-VEGI was confirmed by enzymatic digestion.The expression was mainly in the form of inclusion body.SDS-PAGE electrophoresis and Western blotting revealed a chimeric protein about 23000,with a purity of about 90%.Conclusion:We have successfully constructed the recom- binant plasmid pET30a-VEGI~+ and expressed it in E.coll.And we have obtained high purity of soluble chimeric protein VEGI~+ through affinity chromatography.
4.Behavior problems and family factors in children of off-farm workers of Shanghai
Baolong ZHAO ; Wei JI ; Zhimin ZHAO ; Wenhong CHENG ; Guangping LI ; Meina YOU ; Yong ZHAO ; Min LI
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(12):1073-1076
Objective To survey prevalence of behavior problems in children of off-farm workers and effects of family factors on children' s behavior problems.Methods The study was done in the whole 7 schools for off-farm workers' children in Shanghai Baoshan district,including all 3 ~ 6 grades students of primary school and 1 ~3 grades students of junior high school.Totally 4350 students finished survey (95.4%).Achenbach' s child behavior checklist ( CBCL,parent' s version) and general family questionnaire were used.Results ( 1 ) Prevalence of behavior problems in children was 11.2% ; 12.8% 12 ~ 16 years children had behavior problems( 14.7%boys and 10.4% girls) ; 11.4% of one-child children had behavior problems compared with that of 11.2% child with siblings children ( x2 =0.038,P > 0.05 ).(2) Total score ( 20.64 ± 17.67 ) and factor scores of thought problems,depressed,hyperactivity,delinquent rule-breaking behavior were higher than those of Chinese norm (P <0.05 or P < 0.01 ) among 6 ~ 11 years boy group.Total score ( 17.23 ± 14.73 ) and scores of thought problems,depressed,social problems,hyperactivity,delinquent rule-breaking behavior were higher than those of Chinese norm (P < 0.01 ) among 6 ~ 11 years girl group.Among 12 ~ 16 years boy group,total score (20.44 ± 19.32) and all factor scores of CBCL were higher than those of Chinese norm(P<0.01 ).Total score ( 17.02 ± 16.72) and scores of immature,delinquent rule-breaking behavior,brutality were higher than those of Chinese norm(P < 0.05 or P <0.01 ) among 6 ~ 11 years girl group.(3)Multi-regression analysis showed that poor parent-child relationship was the most important predict factor for higher score of CBCL among all family risk factors (P < 0.05 ).Conclusion Children of the off-farm workers and their families need special mental health care program.
5.Imaging diagnosis of superior ophthalmic vein enlargement associated with orbital arteriovenous malformation
Jin-Wei CHENG ; Rui-Li WEI ; Ji-Ping CAI ; You LI ;
Ophthalmology in China 2006;0(06):-
Objective To investigate the radiologic and angiographic findings of dilated superior ophthalmic vein(SOV)associat- ed with orbital arteriovenous malformations(AVM).Design Retrospective case series.Participants Six cases of orbital AVM patients, with clinical symptoms and signs mimicking a carotid cavernous fistula(CCF).Methods Examinations of the six cases,such as CT, MRI,and selective cerebral angiography,were summarized and analyzed.Main Outcome Measures Images findings and hemodynam- ics.Results All patients were found with dilated SOV on CT and MRI.Additional radiographic findings included proptosis and AVM. All six AVM were located at the orbit,and one of them was also found with AVM in brain.The main draining vein was SOV in all cas- es.The branches contributing to the fistula's blood supply were the middle meningeal artery,the internal maxillary artery,and the oph- thalmic artery.Conclusions Orbital AVM can cause similar clinical signs and radiologic features to those associated with a CCF.How- ever,the bulging cavernous sinus cannot be found in most AVM.Angiography is necessary for definitive management,non-invasive techniques have an important role in pre-angiographic diagnostic decisions.(Ophthalmol CHN,2007,16:395-398)
6.Meta analysis of methylprednisolone pulse therapy for thyroid-associated ophthalmopathy
Jin-Wei CHENG ; Rui-Li WEI ; Ji-Ping CAI ; You LI ;
Ophthalmology in China 2006;0(06):-
Objective To evaluate the efficacy of methylprednisolone pulse therapy in the treatment of thyroid-associated oph- thalmology. Design Meta analysis. Participants Published literatures about methylprednisolone pulse therapy for thyroid-associated ophthalmopathy. Methods A meta-analysis of published randomized clinical trials was used. Published literatures were selected for study based on a computerized search of the literature and a manual search of the bibliographies of relevant papers. Studies meeting se- lected criteria were reviewed systematically by meta-analysis. Data was extracted on the basis of the intention-to-treat principle. Treat- ment effect was measnred as relative risk(RR) of response rates. Main Outcome Measures The effective rate and the relative risk(RR). Results A total of 6 articles were included in this meta-analysis. The RR (95%CI) were 7.50 (1.14, 49.26) when comparing methylpred- nisolone pulse therapy with placebo, 3.33 (0.51, 21.89) when comparing methylprednisolone pulse therapy with surgical decompression, and 1.50 (1.12, 2.00) when comparing methylprednisolone pulse therapy with oral steroids. The combination of methylprednisolone pulse therapy and orbital radiotherapy was significantly effective than oral steroids combined with orbital radiotherapy which pooled RR (95% CI) was 1.42 (1.11, 1.81). Conclusions Methylprednisolone pulse therapy appears to be an effective treatment for thyroid-associated ophthalmology, and seems to have increased effect when given with orbital radiotherapy.
7.Status of patients with allergic rhinitis combined with bronchial asthma in Nanjing and the correlation of clinical characteristics
Fei XUE ; Zeqing LI ; Qiuping WANG ; Xiaofeng XIN ; Manjie JIANG ; You CHENG ; Junfeng JI
Chinese Journal of Tissue Engineering Research 2007;11(43):8809-8812
BACKGROUND: Both Allergic rhinitis and bronchial asthma are allergic diseases of respiratory tract. They frequently coexist in the same patients.OBJECTIVE: To study the incidence of allergic rhinitis in patients with bronchial asthma in Nanjing, and to analyze the correlation of clinical characteristic between allergic rhinitis and bronchial asthma.DESIGN: The questionnaire survey.SETTING: Department of Otorhinolaryngology-Head and Neck Surgery and Department of Respiratory, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA.PARTICIPANTS: A total of 134 patients with bronchial asthma including 73 males and 61 females were selected form Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA from February 2001 to April 2005. Their ages ranged from 3 to 72 years. Severity of asthma of all patients was classified based on Global Initiative for Asthma (GINA). The diagnostic criteria of allergic rhinitis were established referred to the guideline of Haikou Meeting (November, 1997). All patients provided the confirmed consent.METHODS: The questionnaire survey of 134 bronchial asthma patients from the city of Nanjing was conducted. Their age, classification, severity, treatment status, and the family history of all 134 patients were investigated in details. Differences of age, age at onset, and course of disease between rhinitis and asthma of all patients were examined by two independent t tests. The correlation between clinical characteristics of allergic rhinitis and bronchial asthma was analyzed by Spearman rank correlation.MAIN OUTCOME MEASURES : Incidence of allergic rhinitis in patients with bronchial asthma and the correlation of clinical characteristic between allergic rhinitis and bronchial asthma.RESULTS:Of all 134 bronchial asthma patients,82(61.2%,82/134)were accompanied with allergic rhinitis.There were 45 males and 37 females.Of 82 patients with allergic rhinitis accompanying with bronchial asthma,severity was classified into grade Ⅰ (mild pause,n=56),grade Ⅱ (mild persistence,n=21),grade Ⅲ (moderate persistence,n=4),and grade Ⅳ (severe persistence,n=1).In addition,allergic rhinitis was classified into periodicity (n=65) and persistence(n=17).According to the classification and seventy system recommended in ARIA,63 patients had mild symptoms and 19 had moderate and severe symptoms.Severity of allergic rhinitis was positively related to that of bronchial asthma(r=0.689,P<0.01).CONCLUSION: The morbidity of allergic rhinitis combining with bronchial asthma is high in Nanjing, China. While, their classification and severity are highly coherent.
8.Hydrophilicity and adsorptivity of a novel scaffold twice embedded by lecithin and poly-L-lysine to human nasal septum chondrocytes
You CHENG ; Qiuping WANG ; Manjie JIANG ; Kunmin WU ; Fei XUE ; Wei CHEN ; Junfeng JI ; Zeqing LI
Chinese Journal of Tissue Engineering Research 2007;11(9):1786-1789
BACKGROUND:Chitosan is a kind of natural biomaterial and is characterized by great biocompatibility, progressive degeneration and absorption and excellent mechanical property; however, whether it may become an ideal cytoskeleton in the engineering of cartilage tissue or not should be researched further.OBJECTIVE: To observe the hydrophilicity and adsorptivity to human nasal septum chondrocytes and the effect of its function of a novel scaffold made by [poly (dl-lactide-co-glycolide)] (PLGA)/chitosan nonwoven cloth embedded with lecithin (LEC) and poly-L-lysine (PLYS).DESIGN: Blank control study.SETTING: Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out in the Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA from October 2005 to June 2006. Chitosan nonwoven cloth was provided by Hainan Xinlong Company. The mainly technical parameters were detailed as the follows: degree of deacetylation ≥ 90% and relative molecular weight 2-5 × 105. PLGA/chitosan nonwoven cloth scaffold was made in Department of Otolaryngology-Hean and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA and High Polymer Institute of Sun Yat-sen University. The mainly technical parameters were detailed as the follows: mole ratio between monome lactide and glycolide 75:25, porosity 82%-86%, pore diamater 100-300 μm, shear strength 48 MPa, depth 1.5 mm and completely degenerated duration 14-18 weeks. Human nasal septum chondrocytes were the septal cartilage of nose which was derived from operated patients with deflection of nasal septum under sterile condition.METHODS: PLGA/chitosan nonwoven cloth was sheared into pieces with the size of 1.5 cm × 1.5 cm, dipped in 0.01 volume fraction of LEC anhydrous alcohol and 1 g/L PLYS for 6 hours, dealt with ultraviolet radiation after dehydration for 1hour, dipped in 0.75 volume fraction of ethanol for 24 hours, washed with Hanks solution, and incubated for 24 hours. After operations mentioned above, two kinds of novel scaffolds containing various components were obtained, and they were simple scaffold and scaffold embedded with LEC and PLYS. Cells derived from the third generation of human nasal septum chondrocytes were used to make suspension. In addition, cell suspension was grown on those two scaffolds to determine the degree of hydrophilicity through observing diffused degree of cell suspension. Whether cell suspension of human nasal septum chondrocytes was wafted on scaffolds or not were observed under phase contrast microscope so as to determine adsorptivity between cells and scaffolds; meanwhile, growth of cells and production of matrix were also observed.MAIN OUTCOME MEASURES: ① Hydrophilicity of those two kinds of scaffolds (diffused degree of cell suspension of human nasal septum chondrocytes on scaffold) and adsorptivity to cells (whether cell suspension of human nasal septum chondrocytes was wafted on scaffolds or not); ② growth of cells and production of matrix.RESULTS: ① When simple scaffold was put in cell suspension of human nasal septum chondrocytes, cell suspension showing like balls attached to the surface of scaffold, and then, scattered into space of scaffold gradually. During the period of culture, phase contrast microscope indicated that masses of cells attached to the surface of fiber of scaffold.When the petri dish was shaken, cell groups drifted irregularly. The adherent rate was (21±3.7)%. With the cultured time passing by, matrix was not produced. ② When scaffold embedded with LEC and PLYS was put in cell suspension of human nasal septum chondrocytes, cell suspension scattered into space of scaffold rapidly. The adherent rate was (89±5.6)%, which was higher than that of single scaffold group. This suggested that scaffold showed a strong hydrophilicity.Phase contrast microscope indicated that chondrocytes as the form of monome or community were distributed between scaffold and fiber averagely and attached to the surface of fiber of scaffold. When the petri dish was shaken, cell groups did not drift irregularly, and only a few of cells were scarred at the bottom of petri dish. This suggested that scaffold had a strong adsorptivity to cells. One week after culture, matrix showing like cobweb was produced among fibers of scaffold.With the cultured time passing by, matrix was produced abundantly.CONCLUSION: The novel scaffold of PLGA/chitosan nonwoven cloth embedded with LEC and PLYS is characterized by an excellent hydrophilicity and adsorptivity to human nasal septum chondrocytes; meanwhile, it also can secrete matrix.
9.Effects of different modes of one-lung ventilation on hemodynamics in patients undergoing thoracic operation
Ji LI ; Ping CHEN ; Jin GAO ; Shiqi XIANG ; Heng DAI ; Jiyue YOU ; Shengfen TU
Chinese Journal of Anesthesiology 2012;32(7):849-852
Objective To investigate the effects of different modes of one-lung ventilation (OLV) on hemodynamies in the patients undergoing thoracic operation.Methods Forty-five adult patients undergoing thoracic surgery,were randomly allocated into 3 groups based on the modes of OLV used ( n =15 each):intermittent positive pressure ventilation ( IPPV,VT 6-8 ml/kg,RR 10-14 bpm,I:E 1:2) group,IPPV + positive end-expiratory pressure (PEEP) group and IPPV + continuous positive airway pressure (CPAP) group.Double-lumen tube was inserted.Conrrect positioning was verified by fiberoptic bronchoscopy.The patients were mechanically ventilated.PETCO2 was maintained at 35-45 mm Hg.In group IPPV + PEEP,OLV was performed for 30 min with PEEP of 5 cm H2O and then for another 30 min with PEEP of 10 cm H2O.In group IPPV + CPAP,OLV was performed with IPPV in the lung on the ventilated side and with CPAP of 5 cm H2O in the lung on the operated side (for 1 h).MAP,HR,cardiac output (CO),cardiac index ( CI),stroke volume (SV),and stnoke volume index (SVI) were recorded before induction of anesthesia,at 10 min after intubation,at 30 min of two-lung ventilation,at 30 min and 1 h of OLV,and at the end of operation ( T1-6 ).Arterial blood samples were taken at T1,2,4-6 for blood gas analysis.The levels of blood glucose and lactate were measured.Oxygen delivery ( DO2 ) and DO2 index ( DO2I) were calculated.Results Compared with IPPV group,SV,SVI,CO,CI,DO2 and DO2I were significantly decreased at T4,5 ( P < 0.05),and no significant change was found in the levels of blood glucose and lactate in group IPPV + PEEP,and no significant change was found in the parameters mentioned above in group IPPV + CPAP ( P > 0.05).Compared with IPPV + PEEP group,SV,SVI,CO,CI,DO2 and DO2I were significantly increased at T4,5 ( P < 0.05),and no significant change was found in the levels of blood glucose and lactate in group IPPV + CPAP ( P > 0.05).Conclusion It exerts no influence on hemodynamics using OLV with IPPV in the lung on the ventilated side and with CPAP of 5 cm H2O in the lung on the operated side,however,OLV with IPPV + PEEP can result in hemodynamic fluctuation,but the degree of fluctuation is lesser and DO2 can be maintained in the patients undergoing thoracic operation.
10.Correlations between HIV-1 subtypes and changes in CD4+T cell count over time
Kun FANG ; Xiaoshan LI ; You GE ; Yu JI ; Yue SONG ; Guoping DU ; Wenjuan YAN ; Pingmin WEI
Chinese Journal of Microbiology and Immunology 2017;37(6):461-466
Objective To understand the correlations between HIV-1 subtypes and changes in CD4+T cell count over time in patients with different subtypes of HIV-1 infection.Methods A total of 94 patients who were diagnosed with HIV-1 infection in Nanjing and received at least twice CD4+T cell counting test before antiretroviral therapy (ART) were recruited in this study.Descriptive analysis was used to present the rates of CD4+T cell decline for different HIV-1 subtypes.Logistic regression analysis and nonparametric test were conducted to investigate the factors responsible for CD4+T cell decline and to analyze the correlations between the rates of CD4+T cell decline and HIV-1 subtypes.Results The median monthly rate of CD4+T cell decline was-2.20 [interquartile range (IQR):-11.36-2.13] cell/μl.Of all patients,25.5% (24/94) had a significant decline (≥30%) in CD4+T cell count.Compared with the patients infected with CRF01_AE,those infected with CRF07_BC (OR=0.28,95%CI: 1.7-6.5) or other subtypes (OR=0.16,95%CI: 1.0-2.9) had a lower risk of significant decline in CD4+T cell count.In addition,results of the nonparametric test showed that the patients infected with CRF01_AE (M=-21.54,IQR:-30.97——11.92 cell/μl) had a faster CD4+T cell loss than those infected with CRF07_BC (M=-11.26,IQR:-14.06——5.63 cell/μl) (P=0.033).Conclusion HIV-1 subtype is associated with the rate of CD4+T cell decline.It is important to monitor the changes in CD4+T cell count in patients infected with CRF01_AE and to carry out timely ART.