1.Determination of Seven Kinds of Residual Solvents in Bicalutamide by GC
China Pharmacist 2015;(11):2000-2002
Objective:To establish a headspace GC method for the determination of 7 kinds of residual solvents in bicalutamide, including dichloromethane, n-hexane, tetrahydrofuran, ethanol, ether, acetone and ethyl acetate. Methods: The residual solvents in the substance were determined by GC equipped with an FID detector and linked with an Agilent DB-624 capillary column (30. 0 m × 0. 25 mm × 1. 4 m). The inlet temperature was 200℃ and the FID detector temperature was 250℃. The column temperature was raised by program:the initial temperature was 35℃, maintained for 13 min, raised to 180℃ with a rate of 100℃/min, and maintained for 5 min. The carrier gas was nitrogen and the flow rate was 1. 2 ml·min-1. The heated temperature of the headspace oven was 90℃, the heated time lasted 30 min, and the injection volume was 1. 0 ml. The solution medium was dimethyl sulphoxide (DMSO). Results:Each solvent could be completely separated, and the calibration curve of each solvent showed good linear relationship with good accuracy. Conclusion:The method can be applied for the determination of residual solvents in bicalutamide.
3.Generation of spaO-ompA fusion gene of Salmonella paratyphi A and the immunoprotection of expression product of the fusion gene
Jinqin JIANG ; Yifan SUN ; Wenyan YUE ; Jie YAN ; Ping RUAN
Chinese Journal of Microbiology and Immunology 2012;32(2):152-156
Objective To generate the spaO-ompA fusion gene of Salmonella paratyphi A and its prokaryotic expression system,and to determine the immunoprotection of the recombinant expression product rSpaO-OmpA.Methods A flexible peptide sequence was used to link spaO and ompA genes and a prokaryotic expression system of spaO-ompA fusion gene was subsequently generated.SDS-PAGE and Bio-Rad Agarose Image Analyzer were applied to examine the expression as well as the yield of the target recombinant protein rSpaO-OmpA.The antigenicity and immunoreactivity of rSpaO-OmpA were determined using immunodiffusion test,Western Blot assay and micro-Widal's test.By a mouse infection model,the immunoprotection of rSpaO-OmpA against the lethal challenge of S.paratyphi A was determined.In the animal protective test,the recombinant expressed SpaO (rSpaO) and OmpA ( rOmpA ) were used as the controls.Results The generated spaO-ompA fusion gene had 100% nucleotide and amino acid sequence identities compared to the single spaO or ompA gene.The constructed prokaryotic expression system IPTG E.coli BL21DE3pET42a-spaO-ompA expressed the recombinant protein rSpaO-OmpA.rSpaO-OmpA combined with the antiserum against wholecell of S.paratyphi A to present positive hybridization signal and induced specific antibody in the immunized rabbits.Immunization with 100 or 200 μg rSpaO-OmpA contributed 66.7% (8/12) or 83.3% (10/12) immunoprotective rates in mice when the animals were attacked with S.paratyphi A.The immunoprotective rates produced by rSpaO-OmpA were significantly higher than that of equal rSpaO or rOmpA( P<0.05 ).The sera from rSpaO-OmpA immunized mice presented 1∶5-1∶40 agglutination titers to the H antigens of different S.paratyphi species,and 1∶1-1∶16 immunodiffusion titers to rSpaO,rOmpA and rSpaO-OmpA proteins,respectively.Conclusion The artificially fusion antigen,rSpaO-OmpA,has more powerful immunogenicity and immunoprotection that the equal rSpaO or rOmpA.
4.Effects of nano porous beta-tricalcium phosphate/collagen scaffold modified with human bone morphogenetic protein 2 gene on differentiation of MC3T3-E1 cell lines
Qiang RUAN ; Gang ZHAO ; Rui GUO ; Yue XIAO ; Chao LI
Chinese Journal of Tissue Engineering Research 2016;20(38):5657-5663
BACKGROUND:Bone tissue transplantation or osteogenic material fil ing is after used for bone defect repair. To remove autologous bone tissues can lead to additional damage and secondary deformity, therefore, it is extremely urgent to search for a new osteogenic material. OBJECTIVE:To construct the porousβ-tricalcium phosphate (β-TCP)/col agen scaffold modified with human bone morphogenetic protein 2 (hBMP2) gene, and to observe its effects on differentiation of MC3T3-E1 cel lines. METHODS:The porousβ-TCP/col agen scaffold modified with hBMP2 gene was prepared. Then in vitro culture system of MC3T3-E1 cel lines with composite scaffold was established. There were scaffold and plate groups, and each group was divided into two subgroups according to the different concentrations of plasmid. Samples were col ected and observed morphological y by scanning electron microscope and light microscope after complex culture. After 1, 3, 7 and 14 days of induction, calcium nodules were observed through alizarin red staining, the cel cycle was detected by real-time PCR, and expressions ofαI-chain col agen type I gene, Osterix and bone sialoprotein were observed. RESULTS AND CONCLUSION:The number of cel s adhered, differentated and distributed on the composite scaffold was significantly higher than that of the single scaffold (P<0.05). Alizarin red staining and real-time PCR detection showed that the osteogenesis ability of MC3T3-E1 cel lines in the scaffold group was stronger than that in the plate group. To conclude, the porousβ-TCP/col agen scaffold modified with hBMP2 gene is an appropriate candidate for bone defect repair.
5.Clinical significance of neutrophil-lymphocyte ratio in maintenance hemodialysis patients
Yue ZHANG ; Yizhe RUAN ; Qiang HE ; Li WANG
Chinese Journal of Nephrology 2016;32(9):659-664
Objective To explore how neutrophil-lymphocyte ratio (NLR) is related with inflammation and atherosclerosis,and the role of NLR in hospitalization in maintenance hemodialysis (MHD) patients.Methods MHD patients treated in hemodialysis center of Sichuan Provincial People's Hospital from June till November 2013 were enrolled.Patients with severe infection,cardiovascular events and malignant carcinoma were excluded.NLR was determined from complete blood count differential.Clinical parameters such as serum albumin,lipids,intact parathormone,ferritin,C-reactive protein (CRP),25-(OH) vitamin D,interleukin-6 (IL-6) and alkaline phosphatase (ALP) were collected.Pulse wave velocity (PWV) and ankle-brachial index (ABI) were used to evaluate the arterial stiffness.Spearman analysis was used to evaluate the relationship between NLR and these parameters.All patients were divided into low NLR group (NLR≤3.25) and high NLR group (NLR >3.25) on the median NLR,and their differences in these indexes were analyzed.During the one-year follow-up,the reasons and rates of hospitalization and survival were analyzed.Results One hundred and thirteen MHD patients including 58 males and 55 females were enrolled with (69±49) dialysis age and (54± 15) average age.(1) The NLR was significantly correlated with whole blood count (WBC,r=0.538,P< 0.001),ABIL (r=0.201,P=0.033),ABIR (r=0.235,P=0.012) and total cholesterol (TC,r=-0.414,P< 0.001) and low-density lipoprotein cholesterol (LDL-C,r=-0.378,P < 0.001).(2) Low NLR patients had increased TC,LDL-C and IL-6 as compare with high NLR patients,however decreased ABIL and ABIR (all P < 0.05).(3) Forty one patients were hospitalized 63 times during the follow-up period.Annual hospitalization rate was 558/1000 and the mortality rate was 17.7/1000.(4)NLR in patients at least hospitalized once a year was significantly lower than in patients without hospitalization,while ALP was higher (all P < 0.05).Compared with those in other patients,NLR and hemoglobin (Hb) were significantly lower in patients with hospitalization due to infection,while ALP was higher (all P < 0.05).Conclusions NLR is related with WBC,ABI,TC and LDL-C in MHDpatients.Lower NLR may indicate high risk for cardiovascular,atherosclerosis and hospitalization,probably different form non-MHD patients,which needs more studies to verify.
7.Carbon monoxide-releasing molecule CORM-2 protects against renal ischemia-reperfusion injury in mice
Yongle RUAN ; Lu WANG ; Yue ZHAO ; Junxiang WANG ; Song CHEN ; Changsheng MING ; Gang CHEN
Chinese Journal of Organ Transplantation 2013;34(11):685-689
Objective To investigate if the administration of CORM-2 can provide protection against renal ischemia-reperfusion injury (IRI).Method Murine renal ischemia was induced by clamping left renal pedicles for 40 min with vascular micro damps at 32 C,then the contralateral kidney was removed.CORM-2 or vehicle was administered via intravenous infusion 1 h before the onset of ischemia.The blood plasma and renal samples were obtained at 24 h after reperfusion to assess renal function and cellular injury.Plasma Cr and BUN levels,HE and TUNEL were performed to estimate the magnitude of renal damage.Kidneys were retrieved from indicated animals at various time points after renal IRI,and the sections were prepared for histological evaluation.MPO staining procedures were performed to assess the neutrophils infiltration in the renal IRI.Besides,Immunofluorescent stain of TNF-α was performed on the kidneys which were retrieved from indicated animals to determine the production of inflammatory mediators in renal I/R.Results The plasma Cr and BUN were significantly increased at 24 h after reperfusion in IRI control mice,and CORM-2 treatment could markedly diminish the increase of plasma Cr and BUN in mice subjected to I/R.In parallel,histological analysis demonstrated that CORM2 treatment markedly reduced apoptosis of the renal tubular epithelium cells and hemorrhage.IRI caused marked infiltration and accumulation of the MPO-positive neutrophils in renal interstitium.Administration of CORM-2 before ischemia dramatically inhibited neutrophils infiltration as compared with IRI or iCORM-2 group.Furthermore,we confirmed that CORM-2 markedly decreased production of TNF-α.Conclusion Carbon monoxidereleasing molecule CORM-2 could ameliorate inflammation to protect against the renal IRI in mice.
8.Karyotypes analyses in 632 autism spectrum disorder trios
Tianlan LU ; Zhiliu WU ; Yanyan RUAN ; Ang MEIXIJIA ; Weihua YUE ; Jun LI ; Lifang WANG ; Dai ZHANG
Chinese Journal of Nervous and Mental Diseases 2016;42(3):150-155
Objective To detect chromosomal aberrations of autism spectrum disorder (ASD), we performed karyo?types analyses in 632 ASD trios and then investigated whether copy number variants and neurodevelopment related genes are present in the regions of chromosomal aberrations. Methods Karyotypes analyses were performed in 632 ASD trios (1896 individuals). In addition, we investigated whether there were pathogenic copy number variants located in the rele?vant regions of detected aberrant karyotypes by using the database of the International Standards for Cytogenomic Arrays (ISCA) and the Genomic Variation and Phenotype in Humans using Ensembl Resources (DECIPHER) for ASD patients. Results We detected aberrant results in 22 of 632 patients (3.48%) by karyotypes analyses. Of these 22 aberrant karyo?types, 5 were de novo (0.79%), including the duplication, the translocation, karyotypes of Turner syndrome and the addi?tional material with unknown origin. Seventeen children affected with autism had aberrant karyotypes inherited from one of their parents. By using the ISCA and the DECIPHER database, we found that several copy number variants with high pathogenicity were located in 1q25 and 3p24. Further, these copy number variants consisted of several genes related to neurodevelopment such as TNR, ASTN1, and NMNAT2. Conclusion There are a few de novo chromosomal aberrations in some patients affected with ASD. Copy number variants of several pathogenic neurodevelopmental related genes may exist in the regions of chromosomal aberrations. Karyotypes analyses may be applied to explore the genetic etiology in some patients affected with ASD.
9.Association Study of the Disrupted in Schizophrenia 1(DISC1)Gene Polymorphisms with Schizophrenia
Wei SUN ; Jun YAN ; Lifang WANG ; Mei QU ; Yanyan RUAN ; Tianlan LU ; Weihua YUE
Chinese Mental Health Journal 2002;0(08):-
T,P=0.008; Genotype:P=0.031)of DISC1 gene were significantly associated with schizophrenia.The haplotypes constructed by these two markers were significantly associated with schizophrenia,such as AT(?2=7.065,P=0.008,OR=1.42,95%CI=1.10~1.83)and GA(?2=6.009,P=0.014,OR=0.80,95%CI=0.68~0.96).When the subjects examined with the positive and negative syndrome scale(PANSS),the risk haplotype AT was not significantly correlated with positive,negative,excitement,depression and cognitive impairment factors of PANSS. Conclusion:These findings provide further evidence for DISC1 as a predisposing gene involved in schizophrenia in the Chinese Han Population.However,no positive association is found between DISC1 polymorphisms with schizophrenia clinical symptoms.
10.Assessment for health education in plague affected area in Dingbian country, Shaanxi province in 2008
Suo-ping, FAN ; Chun-lai, RUAN ; Yang-xin, SUN ; Yong-jie, YUE
Chinese Journal of Endemiology 2010;29(2):215-217
Objective To improve the knowledge about controlling plague in cadres, masses, and the medical staff in plague affected area in Dinghian county of Shaanxi province and to assess the efforts of health education activities. Methods In 2008, the education activities carded out by the government-related departments were investigated. The awareness of plague control and assessment was obtained through a written survey, questionnaire and interviews on clinic. Results Some education activities were carried out in plague area of Dingbian county, such as training, issuing educational materials, broadcasting plague scientific educational films and arranging knowledge grand prix. The rates of knowing plague clinic, epidemiology, prevention and the "three prohibitions and three alerts to report" were as follows: the cadres were 50.50%(101/200),63.69%(414/650),78.67%(118/150), the masses were 64.71% (66/102),87.91% (269/306),76.47% (78/102) and the medical staff were 64.18% (543/846) ,68.51%(322/470),67.02%(63/94). The passing rates of the cadres, the masses and the medical staff were 70.00% (35/50),92.16% (47/51),74.47% (35/47). Conclusions Health education can strengthen health consciousness of cadres and masses and improve the ability of the medical stalf on controlling sudden epidemic situation. Reinforcing plague control knowledge and training of medical staff are still important for health education in the future.