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

Atrioventricular Block ; etiology ; Cardiac Catheterization ; adverse effects ; Heart Septal Defects, Ventricular ; therapy ; Hematologic Diseases ; etiology ; Hemolysis ; Humans

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

Acetylcysteine ; therapeutic use ; Animals ; Liver Cirrhosis, Experimental ; drug therapy ; Magnesium Compounds ; therapeutic use ; Rats ; Rats, Sprague-Dawley

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

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.

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

OBJECTIVEFrequency, type and clinical implications on protease and reverse transcriptase drug resistance mutations were investigated and phylogenetic analysis in antiretroviral drug-naïve AIDS patients was carried out in Henan province.

METHODS45 plasma samples were separated from the anticoagulatory whole blood, from which reverse transcription-polymerase chain reaction technique was used to amplify the partial pol gene. The sequences were analysed for genotypic antiretroviral resistance and phylogenetic relation through landing the websites http://hivdb.stanford.edu and http://hiv-web.lanl.gov, under BioEdit and DNAClub software.

RESULTSPartial pol sequences of 36 samples were successfully amplified. The major mutation rate of resistance to protease was 8.3% (3/36), including types D30A, V32A, G73C and V82A. Minor mutation rate of resistance was 100%, including types of L63PS (36/36), I93L (35/36), V77IL (34/36), A71IVT (10/36) and D60E (2/36). The mutation rate of resistance to reverse transcriptase was 38.9% (14/36). Mutation-scoring and clinical implication clewed drug resistance rates were 5.6% (2/36) and 22.2% (8/36) to protease inhibitors and reverse transcriptase inhibitors respectively, while 1 sample was potentially low-level resistant to all of the protease inhibitors and 3 samples to part of the reverse transcriptase inhibitors. Phylogenetic analysis revealed that the pol gene of 36 samples were highly homologous and having a near relative to B.US.83.RF ACC M17451. 36 samples seemed to have the same infection source while their resistance mutations were not due to drug-resistant virus infection but to the evolving of virus in vivo.

CONCLUSIONMost of the antiretroviral drug-naïve AIDS patients in Henan province were sensitive to the currently available antiviral medicine, but antiviral treatment must be in accordance with the strict procedure and to keep better adherence, to avoid the epidemics caused by drug-resistant virus.

Acquired Immunodeficiency Syndrome ; genetics ; Adult ; Anti-HIV Agents ; pharmacology ; China ; Drug Resistance, Viral ; genetics ; Female ; Genes, pol ; genetics ; Genotype ; HIV Protease ; genetics ; HIV Protease Inhibitors ; pharmacology ; Humans ; Male ; Mutation ; Phylogeny ; RNA-Directed DNA Polymerase ; genetics ; Reverse Transcriptase Inhibitors ; pharmacology

OBJECTIVETo observe the effect of Compound Zhajin Granule (CZG) on Toll-like re-ceptor 4 (TLR4) signaling pathway in high-fructose corn syrup induced NASH mice.

METHODSThirty 6-week-old male C3H mice were divided into the high fat and high fructose (HFHFr) group (n = 20) and the control group (n = 10) according to body weight. Mice in the HFHFr group ate high fat diet and drank 20% fructose water, while those in the control group ate common diet and drank common water. After 8 weeks mice in the HFHFr group were divided into two group according to body weight, the HFHFr group and the CZG group, 10 in each group. Mice in the CZG group were fed with high fat forage and 20% fructose water, and administered with 50 mL/kg 12. 8% CZG (prepared by hawthorn, Radix Curcumae, Alisma Orientale, Fritillaria Thunbergii, Silybum Marianum, peach seed in the ratio of 3:1.5:1.5:2:1.5:2:1) by gastrogavage. Mice in the HFHFr group were fed in the same way and daily administered with equal volume of distilled water by gastrogavage. Sixteen weeks later all mice were sacrificed. Body weight, liver wet weight, liver function, and lipid metabolism were detected. Pathological changes of liver tissues were assessed by HE staining, oil red O staining, and Masson staining. Expressions of TLR4, myeloid differentiation factor 88 (MyD88), tumor necrosis factor-alpha (TNF-α) were detected using immunohistochemical staining and real-time fluorescent quantitative PCR.

RESULTSBody weight, alanine aminotransferase (ALT), aspartate aminotransferase (AST) were obviously lower in the CZG group than in the HFHFr group (P < 0.05); oil red O stained area and density were decreased more in the CZG group than in the control group. HE staining showed ballooning inflammation was reduced more in the CZG group than in the HFHFr group. Masson staining was negative. Positive rates of TLR4 and MyD88 and mRNA expressions were significantly lower in the CZG group than in the HFHFr group (all P < 0.05).

CONCLUSIONCZG could significantly inhibit TLR4 signaling pathway of liver in NASH mice.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Diet, High-Fat ; Drugs, Chinese Herbal ; pharmacology ; Fructose ; administration & dosage ; adverse effects ; Inflammation ; Lipid Metabolism ; Male ; Mice ; Mice, Inbred C3H ; Myeloid Differentiation Factor 88 ; metabolism ; Non-alcoholic Fatty Liver Disease ; drug therapy ; Signal Transduction ; drug effects ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism