2.Genetic instability detected by flow cytometry: DNA aneuploid and P16 expression in biopsy specimens from lung cancer.
Zong-li SHEN ; Yue-qing ZHU ; Yi-ping ZHUANG ; Zhi-xiang CHENG ; Xiao-liu WU ; Ya-ping WANG
Chinese Journal of Medical Genetics 2007;24(3):322-324
OBJECTIVETo investigate DNA aneuploid and P16 expression in biopsy specimens from lung cancer, and to study genetic instability and the application of flow cytometry in lung cancer pernicious degree diagnosis.
METHODSBlood cells and cancer cells in biopsy specimens were marked simultaneously with anti-CD45 and anti-P16 fluorescent antibody, and the ratio of CD45+ P16+ cells and CD4- P16+ cells was compared. DNA content in biopsy specimens from lung cancer was detected by flow cytometry.
RESULTSAmong the 74 cases of lung cancer, there are 46 cases of DNA aneuploid (62.2%). Thirty-seven cases of lung cancer expressed P16 lowly (50%). Twelve cases of lung cancer only expressed P16 lowly (16.22%), 21 cases of lung cancer only expressed DNA aneuploid (28.38%), and 25 cases not only expressed P16 lowly but also expressed DNA aneuploid (33.78%). Indexes of malign degree, such as P16 low expression or DNA aneuploid could be detected in 58 cases among the 74 cases (78.38%) by flow cytometry.
CONCLUSIONP16 low expression and DNA aneuploid are the indexes of lung cancer malign degree, and flow cytometry can be used to study genetic instability and evaluate biopsy specimens from lung cancer.
Aneuploidy ; Animals ; Biopsy ; Chromosomal Instability ; genetics ; DNA ; genetics ; Female ; Flow Cytometry ; Gene Dosage ; Gene Expression Regulation, Neoplastic ; Genes, p16 ; Humans ; Leukocyte Common Antigens ; genetics ; Lung Neoplasms ; diagnosis ; genetics ; pathology ; Male ; Mice ; Middle Aged
4.Dynamic study on the anti-HBs level of immunized children born to HBsAg-positive and HBsAg-negative mothers in hyperendemic area of hepatitis B.
Jian GONG ; Rong-cheng LI ; Yan-ping LI ; Jin-ye YANG ; Xiu-rong CHEN ; Yi NONG ; Zhao-neng HUANG ; Qiao LI ; Chong-bai LIU ; Hui ZHUANG
Chinese Journal of Epidemiology 2008;29(1):13-16
OBJECTIVETo investigate the dynamic changes of the anti-HBs level among immunized newborn infants born to HBsAg-positive and HBsAg-negative mothers in hyper-endemic area of Hepatitis B.
METHODSInfants who were regularly vaccinated with Hepatitis B vaccine and tested to be anti-HBs positive were divided into two groups according to HBsAg-positive or negative mothers in Long-an, Guangxi. Each subject was followed up 3 times during age 5 to 8. SPRIA was used to test HBsAg, anti-HBs and anti-HBc. Results During the follow-up period, positive rates of anti-HBs in children born to HBsAg-positive mothers ranged between 52.00% and 78.00%, and those with HBsAg-negative mothers was between 43.84% and 54.74%. GMT in two groups was between 55.36 mIU/ml and 95.66 mIU/ml as well as between 39.90 mIU/ml and 65.47 mIU/ml, respectively. There was no statistical significance in both positive rates and GMT between age groups. The anti-HBs level in the follow-up period of children born to HBsAg-positive mothers was higher than that of those born to HBsAg-negative mothers in the same age group. In the age group of 6-8 years with HBsAg-negative mothers, the positive rates in the follow-up period of children with high anti-HBs titers in the primary vaccination were 2.29-2.84 times of those with low titers. The anti-HBs titer of children in a follow-up period was lower than that in the primary vaccination, no matter whether they were born to HBsAg-positive mothers. However, the decline rate of children born to HBsAg-negative mothers was significantly higher than those born to HBsAg-positive mothers (84.91% vs. 61.54%; chi2 = 28.7982, P = 0.000). The incidence rate (25.64%) of a 4-fold increase in antibody titers of children born to HBsAg-positive mothers was significantly higher than that of children born to HBsAg-negative mothers (7.37%) from the primary vaccination to the follow-up period (chi2 = 6.7661, P = 0.009) with was 3.5 times of the latter. Subjects with HBsAg seroconvertion were those with low anti-HBs titers in primary vaccination.
CONCLUSIONThe anti-HBs level decreased slowly in successfully immunized children from age 5 to 8. The chance of natural booster yielded by natural infection increased in immunized children born to HBsAg-positive mothers. The anti-HBs level in the primary vaccination played an important role in prevention of seroconversion of HBsAg.
Child ; Child, Preschool ; Female ; Hepatitis B Antibodies ; blood ; immunology ; Hepatitis B Surface Antigens ; blood ; immunology ; Hepatitis B Vaccines ; immunology ; Humans ; Infant, Newborn ; Male
5.Effect of the complex of casein phosphopeptide and amorphous calcium phosphate on reducing enamel demineralization and promoting remineralization.
Yan CHEN ; Yan ZHOU ; Shi-jian LI ; Ping FANG ; Yi WEI ; Ji-wei TANG
Chinese Journal of Stomatology 2013;48(1):32-36
OBJECTIVETo investigate the effect of the complex of casein phosphopeptide and amorphous calcium phosphate (CPP-ACP) on the demineralization and remineralization of dental enamel in vitro.
METHODSPremolars extracted from patients receiving orthodontic treatment were cut into two slabs, which were embedded and polished. The slabs were randomly divided into non-acid etching group, acid etching group (immersed in 37.5% phosphoric acid for 2 minutes), experimental group A (immersed in 37.5% phosphoric acid for 2 minutes and wiped by CPP-ACP for 3 minutes, then immersed in distilled water for 7 days), experimental group B (immersed in 37.5% Phosphoric acid for 2 minutes and wiped by CPP-ACP for 3 minutes, then immersed in distilled water for 14 days), experimental group C (immersed in 37.5% phosphoric acid for 2 minutes and wiped by CPP-ACP for 3 minutes, then immersed in distilled water for 21 days), experimental group D (immersed in 37.5% phosphoric acid for 2 minutes and wiped by CPP-ACP for 3 minutes, then immersed in distilled water for 28 days), with 6 slabs in each group. The mineral content was determined by scanning electron microscope.
RESULTSThere was a large amount of mineral deposited on enamel surface in experimental group A, B, C, D. The calcium content of experimental group D was higher than those of other 3 experimental groups. The calcium content of experimental group A, B, C, D (66.53 ± 0.63, 67.00 ± 0.49, 67.07 ± 0.24, 67.18 ± 0.50) was higher than that of acid etching group (65.74 ± 0.68) (P < 0.05). There was no significant difference of the calcium content among experimental group A, B, C, D (P > 0.05).
CONCLUSIONSCPP-ACP can reduce enamel demineralization and promote remineralization in vitro.
Caseins ; pharmacology ; Dental Enamel ; drug effects ; Humans ; In Vitro Techniques ; Tooth Demineralization ; Tooth Remineralization
7.Evaluation of bone marrow biopsy and smear examinations in differential diagnosis for patients with pancytopenia by simultaneously compartive study.
Jin-Ying LIN ; Yong-Wei REN ; Wei-Ping ZENG ; Yi-Fang BI
Journal of Experimental Hematology 2004;12(2):228-230
To simultaneously compare and evaluate the examinations of bone marrow smear and trephine biopsy in differential diagnosis and therapeutical effect of pancytopenia patients, the differences between bone marrow smear and trephine biopsy in degree of cellularity, misdiagnosis rate and therapeutical effect for 71 patients with pancytopenia were analyzed. The results showed that the degree of cellularity in bone marrow biopsy for cytologic morphology was higher than that from the smear, but misdiagnosis rate in the biopsy was lower than that in the smear. In conclusion, bone marrow biopsy is necessary to the differential diagnosis and more valuable for evaluation of therapeutical effect and prognosis of pancytopenia patients.
Adolescent
;
Adult
;
Aged
;
Biopsy
;
Bone Marrow
;
pathology
;
Bone Marrow Examination
;
Diagnosis, Differential
;
Diagnostic Errors
;
Female
;
Humans
;
Male
;
Middle Aged
;
Pancytopenia
;
diagnosis
8.Subtype and characterization of gag gene of HIV-1 strains prevalent in some areas of China
Li-Li, CHEN ; Yong-Jian, LIU ; Han-Ping, LI ; Zuo-Yi, BAO ; Dao-Min, ZHUANG ; Si-Yang, LIU ; Zheng, WANG ; Zhang-Li, LAI ; Lin, LI ; Jing-Yun, LI
Bulletin of The Academy of Military Medical Sciences 2009;33(6):505-508
Objective:To investigate the subtype distribution of HIV-1 strains prevalent in four areas of China,and to study the characteristics of gag gene variation and changes in antigen epitopes under the host immune pressures. Methods:The plasma of HIV-1 infected people from Henan, Guangdong, Sichuan and Beijing in China were collected. Virion RNA was extracted directly from plasma after the virion was condensed. The gag gene was amplified by RT-PCR and nested-PCR.Sequences were subtyped by Genotyping Tool software, and phylogenetic analysis of gag gene were performed using the MEGA 4.1 software.The gene distances intra each subtype were calculated by Distance program. The Ks/Ka ratios were calculated using SNAP program. The variation analysis of CTL antigen epitopes restricted by main HLA-Ⅰ specificities in China was performed.Results:Six subtypes or circulating recombinant forms(CRFs)of HIV-1,including B',CRF07_BC,CRF01_AE,B,CRF08_BC and CRF02_AG,were identified in four areas of China.The gene distances intra each subtype were CRF01_AE>B>CRF08_BC> CRF07_BC>B' listed in order of size, meanwhile the order of Ks/Ka ratios was CRF01_AE>B>CRF08_BC>B'>CRF07_BC. Far more diversity of antigen epitopes in P17 region was observed than that in P24.Epitope mutations intra subtypes were CRF01_AE>B>B'>CRF07_BC listed in order of size. Conclusion:Itseems that CRF01_AE is under the strongest immune pressures,and displays the most diversity of gene and variation of epitopes intra subtypes prevalent in China, followed by subtype B, B' and CRF07_BC. The discrepancy of epitope mutations intra the subtypes is significant.
9.Comparative analysis on the complete genome sequence of mumps epidemic strain and mumps vaccine strain S79 isolated in Zhejiang province, China between year 2005 and 2010.
Dong-Yan ZHANG ; Yan FENG ; Shu-Ling ZHONG ; Yi-Yu LU ; Fang-Cheng ZHUANG ; Chang-Ping XU
Chinese Journal of Preventive Medicine 2012;46(3):252-257
OBJECTIVETo compare the differences in the complete genome sequence between mumps epidemic strain and mumps vaccine strain S79 isolated in Zhejiang province.
METHODSA total of 4 mumps epidemic strains, which were separated from Zhejiang province during 2005 to 2010, named as ZJ05-1, ZJ06-3, ZJ08-1 and ZJ10-1 were selected in the study. The complete genome sequences were amplified using RT-PCR. The genetic differences between vaccine strain S79 and other genotype strains were compared; while the genetic-distance was calculated and the evolution was analyzed.
RESULTSThe biggest difference between the 4 epidemic strains and the vaccine strain S79 was found on the membrane associated protein gene; whose average nucleotide differential number was 42.5 +/- 3.0 and the average variant ratio was 13.6%; while the mean amino acid differential number was 12.8 +/- 1.5 and the average variant ratio was 22.4%. The smallest difference among the 4 epidemic strains and the vaccine strain was found in stromatin genes, whose average nucleotide differential number was 73.8 +/- 2.5 and the average variant ratio was 5.9%; while the mean amino acid differential number was 3.0 +/- 0.8 and the average variant ratio was 0.8%. The dn/ds value of the stromatin genes of the 4 epidemic strains reached the highest, as 0.6526; but without any positive pressure (dn/ds < 1, chi2 = 0.87, P > 0.05). There were mutations happened on the known antigen epitope, as 8th amino acid of membrane associated protein genes and on the 336th and 356th amino acid of hemagglutinin/neuraminidase proteins. Compared with the vaccine strain, the glycosylation sites of ZJ05-1, ZJ06-3, ZJ08-1 and ZJ10-1 increased 1, 1, 2 and 2 respectively. The complete amino acid sequence of all strains showed that there were 17 characteristic sites found on the genotype-F mumps strain. Within the complete genome, the genetic-distance between epidemic strains and vaccine strains in Zhejiang province (0.071) was significantly larger than the genetic-distance between strains in Yunnan province (0.013); the difference showing statistical significance (t = 4.14, P < 0.05). Except nucleocapsid protein genes, all the genes shared similar evolution tree.
CONCLUSIONThere were significant differences found in the genes between mumps epidemic strain and mumps vaccine in Zhejiang province.
Amino Acid Sequence ; China ; epidemiology ; Genome, Viral ; Genotype ; Humans ; Molecular Sequence Data ; Mumps ; epidemiology ; genetics ; virology ; Mumps Vaccine ; Mumps virus ; classification ; genetics ; isolation & purification ; Viral Proteins ; genetics
10.Prevalent characteristics in children with hand-foot-mouth disease in the Kunming area in 2010.
Xiao-Hong YANG ; Qian WU ; Bai-Ping WEN ; Hong-Jian DAI ; Yu ZHUANG ; Jun-Yi YANG
Chinese Journal of Contemporary Pediatrics 2012;14(3):192-194
OBJECTIVETo study the prevalent characteristics in children with hand-foot-mouth disease (HFMD) in the Kunming area in 2010.
METHODSThe clinical data of 13286 outpatient and inpatient children with HFMD in Kunming Children's Hospital between January and December, 2010, including 8 death cases, 715 serious cases and 12563 non-serious cases, were retrospectively studied.
RESULTSHuman enterovirus was detected in 8200 children (61.72%). Children infected with EV71 and CoxA16 accounted for 29.49% (2418/8200) and 53.21% (4363/8200), respectively. Seventy-five children (0.91%) were found to have a mixed infection of the two viruses. Other types of human enterovirus were detected in 1344 children (16.39%). There were significant differences in the total positive rate of human enterovirus in the four quarters of the year (P<0.01). The total positive rate in the second quarter represented the highest proportion (71.56%), and the number of patients was also highest, accounting for 52.94% of the total number of patients in the whole year. EV71 infection was common in the serious case group while CoxA16 was found to be the main pathogen in the non-serious case group. Serious cases were common in children under three years old. In the positive EV71 cases, the viral load of EV71 was not statistically different between the death cases, serious and non-serious cases.
CONCLUSIONSIn 2010, children with HFMD in the Kunming area were mainly infected with CoxA16. Serious cases of HFMD were more common in those who were infected with EV71, and the majority of serious infections were suffered by children who were less than three years old. The viral load was not associated with disease severity. The highest morbidity rate was in the second quarter of the year.
Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Enterovirus A, Human ; isolation & purification ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Infant ; Male ; Prevalence ; Retrospective Studies ; Seasons ; Time Factors