2.Relationship between APE1 single nucleotide polymorphism and prognosis of primary small cell carcinoma of esophagus
Yunyao YE ; Gaohua HAN ; Kaijin LU ; Shengbin DAI ; Lixin ZHANG
Clinical Medicine of China 2016;32(1):64-68
Objective To examine the single nucleotide polymorphism(SNP) of apurinic/apyrimidinic endonuclease1 (APE1) in primary small cell carcinoma of esophagus(PSEC) ,then investigate the relationship between these SNPs and the prognosis.Methods Sixty cases first-treated patients with PSEC were recruited, patients with esophageal squamous cell carcinoma (ESCC) and healthy blood donors were recruited as positive and negative controls.APE1 (Asp148Glu) of the patients with PSEC and controls were genotyped by the TaqMan method.Every patient was treated with platinum-based chemotherapy(EP regimen for PSEC and TP regimen for ESCC)and radiotherapy(3D-CRT) ,then every case was followed-up for 2 years.The relationship between these SNPs and the follow-up outcome was analyzed.Results Compared with the ESCC group and control group, APE1 148 pure mutant(Glu/Glu) of PSEC group increased significantly(PSEC group was 40% (12/30), ESCC group was 13.3% (4/30) , control group was 10% (2/20)), the difference was statistically significant (x2 =7.248,P =0.027).According to data of following-up, there was a significant increase in rate of progress (1year:40.0% (12/30) vs 16.7% (5/30), x2 =4.022, P =0.045;2 years: 86.7% (26/30) vs 40.0% (12/30) ,P =0.004) and a significant decrease in survival (33.3% (10/30) vs 76.7% (23/30)) of PSEC compared with ESCC.The SNPs of APE1 Asp148Glu was significantly correlated with frequency of progress, a significant increase was found in rate of progress of the patients with mutant type(Asp/Glu±Glu/Glu) compared with wild genotype(1 year: 50.0%(11/22) ,x2 =3.854,P=0.05;2 years: 81.8% (19/22) ,x2 =10.519,P =0.001) ,the survival of the patients with mutant genotype was significantly lower than wild type (22.7% (5/22) ,x2=10.77,P=0.001).Conclusion The most of polymorphisms of APE1(Asp148Glu) are mutation type in PSEC.Pure mutant genotype (APE1 148Glu/Glu) carry significant enhancement of progression.The polymorphisms of APE1 (Asp148Glu) maybe one of those molecular mechanisms of high frequency of progress and poor prognosis in PSEC.
3.The effect of β-adrenoceptor blockers on G protein and heart function in rats with acute myocardial infarction
Rongsheng XIE ; Lu FU ; Junxian CAO ; Yingnan DAI ; Liqun ZHU ; Ying HAN
Chinese Journal of Geriatrics 2009;28(8):692-695
ObjectiveTo investigate the effect of β-adrenoceptor (β-AR) blockers on G protein and heart function changes in rats with acute myocardial infarction (AMI) MethodsWistar rats with AMI induced by left anterior descending coronary branch ligation were randomly divided into compared with sham operation group. Eight weeks after therapy, hemodynamics was assessed by inserting catheters and the level of G protein was detected by Western blot analysis. ResultsCompared with the sham operation group, systolic blood pressure(SBP), diastolic blood pressure (DBP), left ventricular systolic pressure(LVSP) and left ventrieular pressure maximal rate of rise and fall(±dp/dtmax) in AMI group were significantly decreased, while left ventricular end diastolic pressure (LVEDP) and Gs and Gi protein levels were significantly increased (all P<0.05). Compared with AMI group, LVSP and ± dp/dtmax were increased, but LVEDP and Gi protein level were significantly decreased in metoprolol and carvedilol group. LVEDP and Gi protein level were decreased in carvedilol group compared with metoprolol group. ConclusionsCarvedilol can effectively suppress the change of G protein and improve the heart function after AMI, and the effect is better than that of metoprolol. This may be related with its β2-AR blocking effect.
4.Detection of trisomy 21 by quantitative fluorescent PCR in clinical samples undergoing prenatal diagnosis for hereditary hearing loss
Yanping LU ; Jing CHENG ; Bing HAN ; Longxia WANG ; Pu DAI ; Huijun YUAN ; Yali LI
Chinese Journal of Obstetrics and Gynecology 2011;46(6):427-430
Objective To establish the genetic test technique of trisomy 21 concurrently conducts with prenatal diagnosis for hereditary hearing loss. Methods Fifty-four pregnant women who underwent prenatal diagnosis for hearing loss of their fetuses in Chinese People's Liberation Army General Hospital from March 2009 to May 2010 were enrolled in this study. All probands from the deaf families have confirmed the causative mutation for hearing loss in Genetic Testing Center in Chinese People's Liberation Army General Hospital. The mean age of 54 pregnant women is 31 years at pregnancy of 18 - 26 weeks, 5 cases > pregnancy of 23 weeks, 9 cases ≥ 35 years. All subjects did not conduct the serologic tests for trisomy 21before. Fifteen to twenty ml amniotic fluid was drawn from 49 cases at pregnancy of 18 - 23 weeks and 5 cases > pregnancy of 23 weeks. One to two ml umbilical blood was drawn from 5 cases > pregnancy of 23 weeks. For 9 cases ≥ 35 years, amniotic fluid cell culture and karyotyping analysis were conducted concurrently. A multiple quantitative fluorescent ( QF) PCR and six microsatellite markers were applied to as trisomy 21. Results (1) Fifty-four fetuses were successfully conducted prenatal genetic diagnosis for hearing loss (included GJB2 and SLC26A4). Ten fetuses copied the exactly same genotypes as the probands. The other 44 cases fetuses did not copy the same genotypes as the probands and won't develop hearing loss. The hearing test showed normal hearing for the neonates. (2) All the 54 fetuses were excluded of trisomy 21 by QF-PCR and were verified after birth. Five fetuses with advanced maternal age were performed karyotyping analysis and showed normal. The diagnostic results of QF-PCR can be obtained in 1 - 3 days without misdiagnosed. Conclusions QF-PCR is an efficient, rapid and accurate technique for detection of trisomy 21 without increasing sample amount. It can be used for fetuses who were undertaken hearing loss gene test or other prenatal gene test.
5.Optimization of Forming Process of Luomai Granules by Response Surface Methodology
Junjie XU ; Jinqing LU ; Lijuan WAN ; Jie LIN ; Han TU ; Yi DAI
China Pharmacist 2016;19(9):1665-1669
Objective:To optimize the forming process of Luomai granules. Methods: With the qualified granule yield and the moisture absorption as the indicators, the excipients were screened, and through the study of granulation effect, the proportion of excip-ients was optimized. With the normalized value of qualified granule yield, solubility, hygroscopicity and sensory evaluation as the eval-uation indices, and excipients times, ethanol concentration and amount of citric acid as the main influencing factors with five levels for each, the forming process of Luomai granules was optimized by central combination design-response surface methodology. Results:The option formula for Luomai granules was the follows:the excipients amount was 5-fold of the extract, 85% ethanol was used as the wet-ting agent, and citric acid content was 0. 3%. Conclusion:Optimized by the central combination design-response surface method, Lu-omai granules has such advantages as high qualified yield, low hygroscopicity, good solubility, soft taste and appropriate dosage, sug-gesting the optimized preparation process of Luomai granules is reasonable and feasible.
6.Role of benzodiazepine receptor in amnesic effect of different intravenons anesthetics in mice
Han GAO ; Wei WANG ; Lu WANG ; Jiao WANG ; Zhenguo ZHAI ; Huiming CAI ; Tijun DAI
Chinese Journal of Anesthesiology 2011;31(7):802-804
ObjectiveTo investigate the role of the benzodiazepine receptor in the amnesic effect of propofol,etomicdate and ketamine in mice.MethodsTwo hundred and eighty-eight Kunming mice of both sexes weighing 18-23 g were randomly divided into 9 groups( n =32 each):gruup normal saline + normal saline (group NN); group normal saline+ fat emulsion (group NF); group flumazenil + normal saline (group FN); group normal saline + propofol (group NP) ; group flumazenil + propofol (group FP) ;group nomal saline + etomidate (group NE) ; group flumazenil + etomidate (group FE); group normal saline + ketamine (group NK) and group flumazenil + ketamine (group NK).Normal saline 10 ml/kg was given IP at 10 min before the tests,and normal saline 10 ml/kg,fat emulsion 10 ml/kg,propofol 25 mg/kg,etomidate 3 mg/kg and ketamine 20 mg/kg at 5 min before the tests in groups NN,NF,NP,NE and NK respectively.Flumazenil 1 mg/kg was given IP at 10 min before the tests,and normal saline 10 ml/kg,fat emulsion 10 ml/kg,propofol 25 mg/kg,etomidate 3 mg/kg and ketamine 20 mg/kg at 5 min before the tests in groups NN,NF,NP,NE and NK respectively.Darkness-avoiding test,platform-mounting test and Morris water maze test were performed to assess the cognition function.The latency of response and number of error were recorded in each test.ResultsPropofol,etomidate and ketamine significanfly shortened the duration of latency of response in platform-mounting test as compared with group NN.Etomidate also significantly increased the number of error in platform-mounting test as compared with group NN,while ketamine prolonged the duration of latency of response in Morris water maze test as compared with group NN.Flumazenil significantly counteracted the above action of the 3 intravenous anesthetics.ConclusionBenzodiazepine receptor may play an important role in the amnesic effect induced by propofol,etomidate and ketamine.
7.Study on the promoter hypermethylation of TSHR, P16, and RAS genes in human papillary thyroid carcinoma
Yali DAI ; Jing YE ; Fan ZHANG ; Yuan LIN ; Weiqun PENG ; Donghui LU ; Lingchuan HAN
Chinese Journal of Endocrinology and Metabolism 2010;26(5):381-384
Objective To study the expressions of the tumor suppressor gene TSH receptor( TSHR),P16, and RAS in papillary thyroid carcinoma ( PTC ) , and the correlation between the occurrence of tumor and the aberrant promoter hypermethylation of three tumor suppressor genes. Methods RT-PCR was used to detect the mRNA expression of three tumor suppressor genes in tissues of 50 cases of PTC ,20 cases of nodular goiter,and 12 cases of thyroid adenoma. The promoter methylation status of three tumor suppressor genes was examined by methylation-specific PCR technique( MSP). Gene sequencing was used to test if the hypermethylation existed in the promoter of three tumor suppressor genes. Results In 68.0% (34/50) TSHR gene, 54.0% (27/50) P16 gene, and 60.0% ( 30/50 ) RAS gene in PTCs, hypermethylation in promoter region was detected, the respective results 21.9% (7/32) , 15. 6% (5/32) ,and 31. 3% (10/32) were found in control tissues. The rates of the three genes with promoter hypermethylation in PTC were significantly higher than those in control tissues ( all P<0. 05). The mRNA expressions of TSHR,P16,and RAS were significantly lower in PTC than those in control tissues (0. 41 ± 0.11 vs 0.63±0. 08,0. 51±0. 17 vs 0. 72±0. 22,0. 56±0. 10 vs 0. 67±0. 16, all P<0. 05). The sequencing confirmed that there was CC to TC transmission in the promoters of three tumor suppressor genes. Conclusions The methylation of three tumor suppressor genes in promoter region is a common molecule event and may be involved in the genesis and development of human PTC.
8.Analysis of falling height and trauma in 246 cases of fatal fall
Xuesong LU ; Jianyu DAI ; Feng LI ; Haipeng JIA ; Bo CUI ; Yong ZHANG ; Han ZHANG
Chinese Journal of Forensic Medicine 2016;31(5):480-481,484
Objective To analyze the characteristic of trauma in 246 fatal falls. Methods We collect 246 cases of fatal falls from Pudong district of Shanghai from 2005 to 2014, and then analyze the characteristics of trauma by using different parameters. Results The height in suicides by fall is higher than accidents by fall (25.7m:13.98m). In all fatal falls, head has a higher possibility in suffering trauma than other body regions (75.2%), while neck has a lower possibility (6.5%). The possibility of head trauma in accidents by fall is higher than suicides by fall (89.4%:75.2%). Conclusion Comparing with suicides by fall, victims in accidents by fall have lack of protection from limbs. Besides, the low rate of ifnding neck trauma suggests legal physicians should be more careful in examining this area.
9.Preparation and evaluation of compressive strength and biocompatibilitywith biological calcium citrate cement
Xing DAI ; Qingchun CAI ; Chihua GUO ; Xuezhe HAN ; Wei MA ; Lu YAO
Journal of Xi'an Jiaotong University(Medical Sciences) 2017;38(5):674-678
Objective To produce the biological calcium citrate cement with fresh oyster shells, and investigate its compressive strength and biocompatibility so as to provide the experimental basis for clinical application of the material.Methods The compressive strength of biological calcium citrate cement was measured and its surface morphology was observed by SEM.The calcium release curve and pH value were measured in the simulated body fluid.Last, its biocompatibility was detected by cytotoxicity test.Results Biological calcium citrate cement produced by 0.33mL/g liquid solid ratio had the maximum compressive strength, and the crystal structure of the material was uniform and orderly.The determination of pH value showed that the degradation and absorption of biological calcium citrate cement did not significantly change the pH value of the body fluid.With gradual degradation of the material, the concentration of Ca2+ in the solution increased gradually.Cytotoxicity test showed that this material had good biocompatibility and no cytotoxicity.Conclusion Biological calcium citrate cement possesses strong compressive strength and good biocompatibility, and it can form a microenvironment with low in alkaline and high in calcium.
10.Expression of ksdD Gene Encoding 3-ketosteroid-△~1-dehydrogenase from Arthrobacter simplex in Bacillus subtilis and Analysis of Steroid Transformation
Yu LI ; Fu-Ping LU ; Yi-Han LIU ; Yong-Xin DAI ; Lian-Xiang DU ;
China Biotechnology 2006;0(11):-
To improve 3-ketosteroid-△1-dehydrogenase(KSDH) activity and the transformation level for androst-4-ene-3,17-dione, 3-ketosteroid-△1-dehydrogenase gene(ksdD) from Arthrobacter simplex was cloned into plasmid pWB980 and expressed in B. subtilis WB600 under the control of promoter P43. The molecular weight of expressed enzyme was about 55kDa by SDS-PAGE analysis. The activitities assayed by spectrophotometrical method of intracellular and extracellular soluble enzyme were 110?0.5mU and 15?0.6mU per milligram of protein respectively. The transformation rate of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was 45.3%. Compared with Arthrobacter simplex, the enzyme activity of KSDH expressed in B. subtilis was improved about 30 fold, and the transformation level of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was improved about 10 fold. The recombinant B. subtilis cells used in biotransformation of steroids provided a new way for steroid medicines production.