1.Effects of celecoxib on tumor growth and cell apoptosis in human triple-negative breast cancer xenografts in nude mice
Ling WANG ; Baoen SHAN ; Lihong LIU ; Jie LI ; Bin WANG
Cancer Research and Clinic 2011;23(11):739-742
Objective To evaluate the effects of celecoxib on tumor growth and cell apoptosis in human triple-negative breast cancer (TNBC) xenografts in nude mice.Methods Human TNBC MDA-MB-231 cells were inoculated subcutaneously into BALB/c nude mice.The mice (n=32) were then randomly divided into 4 groups,the control group and the celecoxib group (receiving 25,50,100 mg·kg-1·d-1 respectively).At the end of the study,tumor tissues were collected and tumor volume was measured.Cell apoptosis was determined by flow cytometry (FCM) analysis.NF-κB p65 and pS0 protein levels were measured by immunohistochemistry.Caspase-3 and survivin protein levels were detected by western blotting.Results celecoxib at dose of 25,50 and 100 mg·kg-1·d-1 inhibited the tumor growth significantly,compared with the control group.FCM results showed that apoptotic rates were (13.58±3.16) % and (21.91±4.75) % in moderate and high dose of celecoxib-treated group,compared with (3.15±1.73) % in control group (t =6.736,P < 0.05;t =12.151,P < 0.05).p65 expressions were 79.3 %,46.7 % and 23.9 % in low,moderate and high dose of celecoxib-treated group,compared with 89.7 % in control group (x2 =3.312,P < 0.05; x2 =10.785,P < 0.05;x2 =15.900,P < 0.05).Besides,western blotting analysis demonstrated that celecoxib significantly downregulated survivin expression,while upregulated the active form of caspase-3 expression.Conclusion Celecoxib could suppress TNBC tumor growth and induce cell apoptosis,which might be partially associated with inactivation of p65 and downregulation of survivin.
2.Construction of Nfic gene 3′UTR dual luciferase reporter vector and targeting verification between Nfic and miR-20a
Shan WANG ; Xiaoxia LI ; Jie ZHOU ; Baoli WANG
Tianjin Medical Journal 2016;44(9):1065-1068
Objective To construct a luciferase reporter vector containing the 3′untranslated region (3′UTR) of nuclear factor I-C (nuclear factor I-C, Nfic), and apply dual luciferase reporter gene system to determine the association between microRNA-20a (miR-20a) and its potential target gene Nfic. Methods The potential complementary binding sites of miR-20a and Nfic were predicted by Targetscan. The 3′UTR of Nfic fragment amplified by PCR was cloned into luciferase reporter vector MIR- Report Luciferase. The luciferase reporters containing 3′ UTR of Nfic and miR- 20 mimics (experimental group) or NC mimics (control group) were co-transfected into 293-AD cells. Cells were collected, and then dual-luciferase reporter assay was performed to detect the luciferase activity of the two groups of cells, consequently the relationship between miR-20a and Nfic was identified. The miR-20a mimics and NC mimics were transfected into marrow stromal cell line ST2 respectively. The total cell lysates were collected, and the expression level of NFIC was detected by Western blotting assay. Results Results of double enzyme digestion and DNA sequencing showed that sequence of luciferase reporter vector was correct. miR-20a specificity bounded to Nfic 3′UTR and inhibited the luciferase activity of the reporter construct (P<0.05). Western blotting assay showed that the NFIC protein level was obviously down-regulated in ST2 cells after the transfection of miR-20a mimics compared with that of control. Conclusion The luciferase reporter vector containing the 3′UTR of Nfic is constructed successfully, which confirms that miR-20a can direct effect on Nfic3′UTR and repress its luciferase activity.
3.Progress of molecular pathology in pediatric brain tumor.
Ya-jie WANG ; Yue-shan PIAO ; De-hong LU
Chinese Journal of Pathology 2011;40(3):206-208
Astrocytoma
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genetics
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metabolism
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pathology
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Brain Neoplasms
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classification
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genetics
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metabolism
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pathology
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Child
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Child, Preschool
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Chromosomal Proteins, Non-Histone
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genetics
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metabolism
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Chromosome Deletion
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Cyclin-Dependent Kinase Inhibitor p16
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genetics
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metabolism
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DNA-Binding Proteins
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genetics
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metabolism
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Ependymoma
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genetics
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metabolism
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pathology
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Hedgehog Proteins
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genetics
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metabolism
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Humans
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Medulloblastoma
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classification
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genetics
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metabolism
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pathology
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Proto-Oncogene Proteins B-raf
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genetics
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metabolism
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Rhabdoid Tumor
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genetics
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metabolism
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pathology
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SMARCB1 Protein
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Signal Transduction
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Transcription Factors
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genetics
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metabolism
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Wnt Proteins
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metabolism
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beta Catenin
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genetics
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metabolism
4.EML4-ALK and EGFR mutation status and survival analysis in Uygur with stageⅣNSCLC
Qiang WANG ; Qiao ZHANG ; Yanzhen CAO ; Jie TAO ; Li SHAN
Tianjin Medical Journal 2016;44(2):200-204
Objective To investigate the relationship between the echinoderm microtubule associated protein like 4-anaplastic lymphoma kinase (EML4-ALK) and epithelial growth factor receptor (EGFR) mutation status and overall survival (OS) in Uygur patients with stageⅣnon-small cell lung cancer (NSCLC) who did not accept tyrosine kinase inhibitor treat-ment. Methods Totally 97 tissue samples were collected from Uygur patients with stageⅣNSCLC who did not accept tyro-sine kinase inhibitor treatment. EML4-ALK fusion gene and EGFR mutation status were detected by using FISH and ARMS methods. The survival rates were analysed. Results In 97 tissue samples, EML4-ALK fusion genes were found in 6 (6.2%) samples, EGFR mutations were found in 26 (26.8%) samples. The survival analysis showed that there was no significant dif-ference in OS between EML4-ALK fusion gene group and no EML4-ALK fusion gene group (P=0.941). There was no signifi-cant difference in OS between EGFR mutation group and wild-type EGFR group (P=0.607). The values of median OS were 17.7 months, 17.3 months and 16.2 months for EGFR mutant group, EML4-ALK positive group and EML4-ALK negative+EGFR wild-type group, and thre was no significant difference between them (P=0.915). Conclusion Excluding the thera-peutic influence in TKIs, EML4-ALK fusion gene and EGFR mutation status of tumor tissue can not be used as an indepen-dent factor in assessing the prognosis in Uygur patients with stageⅣNSCLC.
5.Preparation and Identification of Recombinant Vasoactive Intestinal Peptide
Shan-Shan XIE ; Rong-Jie YU ; Le ZENG ; Juan LI ; Jing-Jing WANG ; An HONG ;
China Biotechnology 2006;0(11):-
In order to prepare the recombinant vasoactive intestinal peptide (VIP) using intein mediated rapid purification system,the cDNA encoding the recombinant VIP was designed and synthesized according to the preference of E.coli,and then was cloned into the expression vector PTWIN. The recombinant plasmid PTWIN-VIP was transformed into expression host E.coli strain ER2566.The fusion protein consisting of the recombinant VIP,intein and chitin binding domain was expressed and purified by chitin affinity chromatography. The target peptide was released from the fusion protein by changing the temperature and the pH of the cleavage buffer. The molecular weight of the recombinant VIP was determined by the mass spectrometry and the results was conformity with the theoretical value. The preliminary bioactivity assay indicated that the recombinant VIP decreased the serum resistin levels significantly in LPS-induced acute inflammation. The preparation and the characterization of anti-inflammatory effects of the recombinant VIP layed the foundation for its further application.
6.Relationship between alexithymia and impact of event of prison male offenders
Lina LI ; Yi WANG ; Shan WANG ; Ying WANG ; Yongfang GUO ; Xiaomei CHEN ; Jie YUAN
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(5):456-458
ObjectiveTo explore the relationship between the alexithymia and impact of event of the prison male offenders.Methods267 male inmates in one prison of Tangshan and 99 men from students and general workers have been tested with Toronto Alexithymia Scale (TAS-26) and Impact of Event Scale-Revisod( IESR).Results ① The alexithymia scores of prison male offenders were more higher except for the factor Ⅰ (Prion:18.62 ± 3.61 ; general:18.00 ± 3.57 ) and factor Ⅲ ( prion:14.38 ± 3.06 ;general:14.96 ± 3.34).And there were significant differences in factor Ⅱ ( Prison:22.26 ± 3.96 ; general:19.72 ± 3.34 ),factor Ⅳ ( prison:22.38 ± 4.20;general:21.03 ± 4.03 ) and the total score (prison:77.65 ±7.22;general:73.71 ±8.32) (P<0.01).② Sex offenders'alexithymia degree was the heaviest ( 79.90 ± 5.00 ) and violence perpetrator' degree was the lightest (77.55 ±7.43),but there was no significant difference among the three subjects(P>0.05).There were significant differences in the evade dimension and the total score in impact of evens between the three different type prison male offenders.③There were negative eorrehtions between factor Ⅲ and factor Ⅳ of alexithymia and every dimension of impact of events( r =-0.14 ~ -0.22,P< 0.05),there were positive correlations between factor Ⅰ and factor Ⅱ of alexithymia and every dimension of impact of events( r =0.14 ~ 0.43,P < 0.05 ).ConclnsionThe degree of alexithymia in prison male offenders is higher than the general men.The violent male offenders are served more impact of events.The higher the degree of alexithymia of the prison male offenders,the more impaction of the event they served.
7.Genetic polymorphisms of 19 STR loci in Shandong Han population.
Mao-Xiui ZHANG ; Shu-Yi HAN ; Hong-Mei GAO ; Shan-Hui SUN ; Dong-Jie XIAO ; Yang LIU ; Yun-Shan WANG
Journal of Forensic Medicine 2013;29(6):440-446
OBJECTIVE:
To investigate the genetic polymorphisms of 19 STR Loci in Shandong Han population in order to provide the genetic data for paternity testing.
METHODS:
The genotypes of 205 unrelated individuals in Shandong Han population were typed by Goldeneye 20A kit to get the allele frequencies and population genetic parameters of 19 STR loci. Four kits, Identifiler kit, SinoFiler kit, PowerPlex 16 kit, and Goldeneye 20A kit, were compared with each other and used in the analysis of a special paternity test case.
RESULTS:
The population genetic parameters of 19 STR loci in Shandong Han Population were obtained. The cumulative discrimination power (CDP) and cumulative probability of exclusion (CPE) ranked from high to low were Goldeneye 20A kit, SinoFiler kit, PowerPlex 16 kit and Identifiler kit, respectively. As duo case, the result of the real case showed that Identifiler kit had no excluding loci, and none of the SinoFiler kit, PowerPlex 16 kit or Goldeneye 20A kit could exclude fatherhood.
CONCLUSION
Compared with Identifiler kit, SinoFiler kit, and PowerPlex 16 kit, Goldeneye 20A kit shows the higher efficiency than the others, but is not completely satisfied for duo cases.
Asian People/genetics*
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China
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Forensic Genetics/methods*
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Gene Frequency
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Genetic Loci/genetics*
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Genetics, Population
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Genotype
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Humans
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Male
;
Microsatellite Repeats
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Paternity
;
Polymorphism, Genetic/genetics*
8.Study of universal salt iodization on child intellectual quotient development in Hainan province
Hong-mei, WANG ; Ming, QIAN ; Hui-jie, DONG ; Shan-qing, WANG ; Ying-di, SU
Chinese Journal of Endemiology 2010;29(1):82-85
Objective To estimate the impact of universal salt iodization on child intelligence quotient (IQ) development in Hainan province. Methods In this is a cross-section study from September in 2008 to April in 2009,the observation group was historical iodine deficiency ward which includes 2 counties and 6 townships,and the control group was non-historical ward which includes 6 counties and 9 townships. Comparison of child IQ distribution according to different geographical housing position(plain,mountainous area,coast),age and sex based on the result of urine iodine examination and the IQ test of children between 8 and 10 years old. Results The median of urinary iodine of children in the observation group was 196.2μg/L which was 2.13 times of the urinary iodine median in the control group(91.9μg/L). The average IQ of children in the observation group was 94.7 which was 8.5 higher than the control group(86.2) ; IQ≤69 rate in this group was 7.7%(91/1179),and it was 8.9 percentage point lower than the control group[16.6%(253/1520)]; IQ≥ 110 rate of the group was 18.3%(216/1179),and it was 10.0 percentage point higher than the control group[8.3%(126/1520)]. The average IQ of children living in mountainous area(83.1) was the lowest in the control group. It was 3.5 and 5.1 lower than that of the children living in the plain(86.6) and coastal area(88.2) respectively; the IQ≤69 rate of children living in mountainous area [20.5%(91/443)]was the highest,and it was 3.8 and 7.1 percentage point higher than that of the children living in the plain [16.7% (89/533)]and coastal areas [13.4% (73/544)]respectively. The average IQ of children aged 8 (97.4) was similar to those aged 9(95.9) in the observation group which was 6.8 and 5.3 higher than that of the children aged 10(90.6) in the same group respectively; However,the average IQ of children aged 8,9 and 10 was close in the control group(86.8,86.3 and 85.6). The average IQ of boys(96.2) was 3.1 higher than that of the girls(93.1),and their IQ≤69 rate[6.3%(37/590)]was 2.9 percentage point lower than that of the girls[9.2% (54/589)]in the observation group. On the other hand,the average IQ(87.2) of boys was 2.1 higher than that of the girls(85.1),and IQ≤69 rate[14.5%(114/787)]was 4.5 percentage point lower than that of the girls [19.0%(139/733)]in the control group. The average IQ of children with different housing geographical position,age and sex in observation group was 5.0-12.4 higher than that of the control group; their IQ≤69 rate was 7.7-13.2 percentage point lower than that of the control group; their IQ≥110 rate was 5.6-13.0 percentage point higher than that of the control group. Conclusions Supplementing salt iodization can improve child intelligence. Supplementing iodine can increase the child IQ and reduce the child mental retardation.
9.Expression of nestin and Ki-67 in malignant peripheral nerve sheath tumors and its significance
Shan LU ; Hua WANG ; Yuping WANG ; Yu YANG ; Yanfeng ZHONG ; Jie ZHENG
Cancer Research and Clinic 2012;24(4):246-249
Objective To explore the expression of nestin and Ki-67 in malignant peripheral nerve sheath tumors (MPNST) and its significance in the differential diagnosis. Methods Immunohistochemical technique (SP) was used to detect the expression of nestin and Ki-67 in 42 cases of MPNST and 24 cases of benign peripheral nerve tumor.Results Total expression of nestin was found in 95.2 %(40/42) of MPNST.Strong expression of nestin was detected more frequent in MPNST compared to benign peripheral nerve tumors [40.5 %(17/42) versus 4.2 %(1/24),x2 =8.403,P =0.004].Ki-67 labeling index in MPNST varied from 1%-70 %.However,greater than 3 % labeling index of Ki-67 staining was observed in 64.3 %(27 / 42) of MPNST while none of the 24 benign tumors had nuclear staining exceeding 3 %. The higher Ki-67 labeling index showed significant differences between the two groups (x2 =23.518,P =0.000).Conclusion Nestin and Ki-67 are useful markers in distinguishing MPNST from benign tumors.
10.Antimicrobial and disinfectant resistance of pathogens isolated from hospi-tal environmental inanimate surfaces and hands of health care workers
Huiping WANG ; Hongjiang ZHANG ; Qi DONG ; Jie LIU ; Shan DUAN ; Junqi GE ; Zhonghua WANG
Chinese Journal of Infection Control 2016;15(12):921-925
Objective To investigate the types,antimicrobial resistance,and disinfectant resistance of pathogens isolated from hospital environmental inanimate surfaces and hands of health care workers (HCWs).Methods Pathogens isolated from hospital environmental inanimate surfaces and hands of HCWs in intensive care units and general wards in 16 hospitals in Beijing were performed bacterial identification,antimicrobial susceptibility testing,and disinfectant re-sistance testing. The carriage of antimicrobial resistance genes and disinfectant genes in pathogens were also detec-ted.Results A total of 979 specimens were collected from inanimate surfaces and hands of HCWs in 16 hospitals,75 (7.66% )pathogenic strains were isolated,78.67% of which were gram-negative bacilli. The top 3 pathogens were Pseud-omonasaeruginosa (P.aeruginosa,n= 24),Enterobactercloacae (E. cloacae,n= 14),and Klebsiella pneumoniae (K. pneumoniae,n= 4 ). One P. aeruginosa strain was resistant to aztreonam,gentamycin,tobramycin,ciprofloxacin,and levofloxacin;One E. cloacae strain was resistant to piperacillin,7 strains were resistant to nitrofurantoin;4 K. pneumoni-ae strains were all resistant to piperacillin,2 were resistant to cephalosporins,and 1 was resistant meropenem. P. aerugi-nosahad7drug-resistantgenes,positiverateofmirwas100.00% ;E.cloacaehad4drug-resistantgenes,positiveratesof tem 1and shv were both 100.00% ;K. pneumoniae had 5 drug-resistant genes,positive rates of shv and mir were both 100.00% . The resistant rates of P. aeruginosa and E. cloacae to chlorhexidine gluconate were 4.17% and 57.14% re-spectively,to trichloroisocyanuric acid were both 50.00% ,positive rates of drug-resistant genes (qacE△1-sul 1)were 79. 17% and 57.14% respectively;K. pneumoniae had no resistance to two kinds of disinfectant,dug-resistance gene was not found.Conclusion Multiple common pathogens which can cause healthcare-associated infection exist in hospital environ-mental inanimate surfaces and hands of HCWs,which are dominated by gram-negative bacilli,pathogens had resistance to antimicrobial agents and disinfectant in different degrees.