OBJECTIVETo explore the value of detection of adenomatous polyposis coli (APC) gene and K-ras gene mutations in fecal and blood samples in colorectal neoplasm screening.

METHODSFrom 84 subjects undergoing colonoscopic examination (including 31 with colorectal carcinoma, 26 with colorectal adenoma, and 27 healthy subjects) between October, 2003 and March, 2004, 5 ml of heparinized peripheral blood and 3-5 g fecal specimens were collected. The DNA was extracted from the specimens for detecting the mutation of APC and K-ras gene using polymerase chain reaction-single strand conformation polymorphism and the results were analyzed statistically.

RESULTS AND CONCLUSION(1) The incidence of APC gene mutation was 41.9% and 57.7% in the plasma, and 34.8% and 26.8% in the fecal specimens of colorectal carcinoma patients and adenoma patients, respectively, both higher than that in normal subjects (P<0.05), suggesting high consistency between fecal and plasma APC gene mutation detection (K=0.811, P<0.05). (2) The incidence of plasma K-ras mutation was 48.4% in colorectal carcinoma patients, 3.8% in adenoma patients and 0% in normal control subjects, and in the feces, the incidences were 54.8%, 7.7% and 11.1%, respectively. The mutation rate was higher in carcinoma patients than in adenoma patients and normal subjects (P<0.05). Fecal K-ras gene mutation detection was consistent with plasma detection (K=0.662, P=0.000). (3) APC gene mutation detection showed a low sensitivity and specificity in the diagnosis of colorectal carcinoma, but K-ras mutation detection showed a high specificity. The diagnostic sensitivity increased when combining APC and K-ras gene detection in the plasma or fecal specimens, but there was no evidence to suggest that APC and K-ras mutation detection in the plasma could be better than detection in the feces. (4) For colorectal carcinoma, APC gene mutation is associated with lymphoid node metastasis, but not with the patient's gender, age, tumor location, differentiation, distant organ metastasis or CEA level (P>0.05), and the mutation of K-ras gene is related to the degree of tumor differentiation.

Adenomatous Polyposis Coli ; blood ; genetics ; Case-Control Studies ; Colorectal Neoplasms ; blood ; diagnosis ; genetics ; pathology ; DNA ; blood ; Feces ; Female ; Genes, ras ; genetics ; Humans ; Male ; Middle Aged ; Mutation

OBJECTIVETo explore the factors influencing failure of an immunization to interrupt perinatal (mother-to-child) transmission of hepatitis B virus (HBV).

METHODSBetween June 2006 and March 2010, a total of 1355 pregnant women testing positive for the hepatitis B surface antigen (HBsAg), at gestational weeks 20 to 42, and without use of antiviral or immunomodulatory drugs during the pregnancy were prospectively recruited to the study. The mothers were given a choice of receiving hepatitis B immunoglobulin (HBIG; three 200 IU intramuscular injections give at four-week intervals starting from gestation week 28) or not. All neonates (1360, including five sets of twins) received hepatitis B vaccine (10 mug) plus HBIG (200 IU) combined immunization within 24 h of birth, as early as possible. Peripheral venous blood samples were collected from the neonates within 24 h of birth and at 7 and 12 months of age for detection of HBV markers, including hepatitis B e antigen (HBeAg) and HBV DNA. The infants were classified according to HBV perinatal transmission status (infection group and non-infection group) and various factors (maternal-related: age, gravidity, parity; pregnancy/birth-related: threatened premature labor, complications; neonate-related: sex, birth weight, apgar score) were compared between the two groups by using non-conditional logistic regression analysis to determine their potential influence on failure of immunization to inhibit transmission.

RESULTSAfter 12 months of follow-up, 1.54% (21/1360) of the neonates had presented with HBV infection. Analysis of the HBV-infected neonates revealed differences in infection rates between neonates born to mothers with HBIG injection (2.22% vs. without HBIG injection: 1.11%, P less than 0.05) and caesarean section (1.35% vs. vaginal delivery: 1.73%) but neither reached statistical significance (P less than 0.05); only the practice of breastfeeding showed a significant difference for infection rate, with neonates fed artificial formula having higher infection rate (3.13%) than the breastfed neonates (0.27%, P less than 0.05). The neonate HBV infection rate was also significantly higher for neonates born to HBeAg-positive mothers (4.44% vs. HBeAg-negative mothers: 0%, P less than 0.05) and HBV DNA-positive mothers (3.13% vs. HBV DNA-negative mothers: 0%, P less than 0.05). When the mothers were stratified by serum level of HBV DNA, there was a significant difference in HBV-infected neonates born to mothers with more than or equal to 1*10(7) IU/ml(6.01% vs. 10(3)-10(6) IU/ml: 0.56% and less than 1*10(3) IU/ml: 0%, both P less than 0.05). Logistic regression analysis indicated that the independent risk factors for HBV perinatal transmission despite immunization were maternal serum HBeAg-positive status (relative risk (RR)=31.74, 95% confidence interval (CI): 3.88-259.38) and maternal HBV DNA of ≥ 10⁷ copies/mL (RR=22.58, 95% CI: 4.75-107.40).

CONCLUSIONFailure of vaccine plus HBIG to interrupt mother-to-child transmission of HBV is influenced by maternal serum HBeAg-positive status and maternal HBV DNA of ≥10⁷ copies/mL.

Adult ; DNA, Viral ; blood ; Female ; Hepatitis B ; prevention & control ; transmission ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B Vaccines ; therapeutic use ; Hepatitis B virus ; Humans ; Immunoglobulins ; therapeutic use ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; prevention & control ; Pregnancy ; Pregnancy Complications, Infectious ; prevention & control ; virology ; Pregnancy Trimester, Second ; Pregnancy Trimester, Third ; Prospective Studies ; Risk Factors ; Viral Load

OBJECTIVETo investigate the risk factors of intrauterine hepatitis B virus (HBV) infection and the impact of HBV DNA on the infection.

METHODSThe serum levels of HBsAg, HbsAb, HBeAg, HBeAb, HBcAb and HBV DNA were determined in blood samples from 230 HBsAg-positive pregnant women and their newborns by enzyme-linked immunosorbent assay (ELISA) and fluorescence quantitative PCR (FQ-PCR), respectively. The newborns acquiring HBV infection via intrauterine transmission were selected as the case group and others as the control group. The risk factors for intrauterine HBV infection were analyzed by non-conditional logistic regression model.

RESULTSSix infants were found to be HBsAg-positive, and 18 HBV DNA-positive, and 3 of them were positive for both HBsAg and HBV DNA. The rate of intrauterine HBV infection was 9.6% (22/230). The grade of HBV DNA level was identified as the only risk factor of intrauterine HBV infection by non-conditional logistic regression model, with odds ratio (OR) of 1.57 (95% confidence interval 1.12-2.21). Of the 119 pregnant women positive for HBV DNA, 18 were diagnosed as having intrauterine HBV infection, and the likeliness of the infection significantly increased for a maternal serum HBV DNA level > or =10(7) copies/ml (chi(2)=7.92, P<0.05).

CONCLUSIONThe grade of serum HBV DNA level is the predominant risk factor for intrauterine HBV infection in pregnant women, and for those with serum HBV DNA lever > or =10(7) copies/ml, the chance for intrauterine HBV infection can be significantly increased.

DNA, Viral ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Hepatitis B ; transmission ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Logistic Models ; Odds Ratio ; Polymerase Chain Reaction ; Pregnancy ; Pregnancy Complications, Infectious ; blood ; virology ; Risk Factors

OBJECTIVETo detect the expression of EGFR and p-ERK in nasopharyngeal carcinoma (NPC) and investigate their clinical significance.

METHODSImmunohistochemistry LSAB method was adopted to detect the expression of EGFR and p-ERK. Statistical analysis was performed using SPSS statistical software package (10.0) to correlate their expression with clinical characteristics and prognosis.

RESULTSPositive staining for EGFR was observed in 39 of 55 cases (70.9%). The EGFR expression was correlated with clinical stage and gender. EGFR expression was correlated with poorer overall survival (OS) and shorter time to progression (TTP). Positive staining for p-ERK was observed in 29 of 55 cases (52.7%). There was a statistically significant association between positive p-ERK expression and advanced clinical stage. Positive p-ERK expression was correlated with poorer OS, disease-free survival (DFS) and TTP. EGFR expression was correlated with the expression of p-ERK. On multivariate analysis, age over 50 years was an independent poor prognostic factor for NPC. Both EGFR and p-ERK were not independent prognostic factors for NPC.

CONCLUSIONExpressions of EGFR and p-ERK are detected in NPC. Their abnormally high expression signifies poor prognosis in NPC patients.

Age Factors ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Mitogen-Activated Protein Kinases ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Staging ; Proportional Hazards Models ; Receptor, Epidermal Growth Factor ; metabolism ; Sex Factors ; Survival Rate

OBJECTIVETo purify Methamidophos (Met) monoclonal antibodies with two methods and compare immune activity of purified antibodies.

METHODCaprylic acid ammonium sulphate precipition (CAASP) method and Sepharose protein-A (SPA) affinity chromatography method were used to purify Met monoclonal antibodies, UV spectrum scanning was used to determine protein content and recovery of purified antibodies, sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze the purity of purified antibodies, and enzyme-linked immunosorbent assay (ELISA) was used to determine immune activity of purified antibodies.

RESULTSAntibody protein content and recovery rate with CAASP method were 7.62 mg/mL and 8.05% respectively, antibody protein content and recovery rate with SPA method were 6.45 mg/mL and 5.52% respectively. Purity of antibodies purified by SPA method was higher than that by CAASP method. The half-maximal inhibition concentration (IC50) of antibodies purified by SPA to Met was 181.26 microg/mL, and the linear working range and the limit of quantification (LOD) were 2.43-3896.01 microg/mL and 1.03 microg/mL, respectively. The IC50 of antibodies purified by CAASP to Met was 352.82 microg/mL, and the linear working range and LOD were 10.91-11412.29 microg/mL and 3.42 microg/mL, respectively.

CONCLUSIONAntibodies purified by SPA method are better than those by CAASP method, and Met monoclonal antibodies purified by SPA method can be used to prepare gold-labelled testing paper for analyzing Met residue in vegetable and drink water.

Antibodies, Monoclonal ; immunology ; Chromatography, Affinity ; Chromatography, Agarose ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Food Contamination ; Fruit ; Insecticides ; immunology ; Organothiophosphorus Compounds ; immunology ; Pesticide Residues ; analysis ; immunology ; Vegetables

Adolescent ; Adult ; Aged ; Arrhythmias, Cardiac ; diagnosis ; etiology ; Child ; Child, Preschool ; Ebstein Anomaly ; complications ; pathology ; Humans ; Infant ; Middle Aged ; Young Adult

Objective To investigate the relationship between serum thyrotrophin(TSH)and dyslipidemia in subclinical hypothyroid and euthyroid subjects. Methods An epidemiological study on diabetes and thyroid diseases was performed in Dadong community, Shenyang city, in 2007. 110 subjects with subclinical hypothyroidism(SCH)and 1 240 euthyroid subjects were enrolled in the study. Neither history of thyroid diseases nor administration of thyroid-related and lipid-regulating medicines were reported in these subjects. The levels of serum thyroid hormones, lipids, fasting plasma glucose(FPG), and insulin were determined. Results (1)Patients with SCH had significantly lower HDL-C levels than those who were euthyroid.(2)According to the guideline of treatment of adult dyslipidemia in China, the lipid profiles were each categorized. Mean TSH levels were higher in subjects in the dyslipidemia subclass than subjects in the normal subclass. The differences were significant in high LDL-C subclass in overweight individuals. In euthyroid overweight women, mean TSH levels were significantly higher in high LDL-C subclass. In the euthyroid population, TSH was positively associated with total cholesterol in overweight population. The association was not modified by the homestasis model assessment for insulin resistance(HOMA-IR)values.(3)TSH was associated positively with serum triglycerides and negatively with serum HDL-C in women. TSH was positively associated with total cholesterol in overweight population and positively associated with total cholesterol and LDL-C in overweight women after adjustment for age, sex, and body mass index. Conclusion Raised serum TSH seems to be a risk factor of dyslipidemia in subclinical hypothyroid and euthyroid subjects, which is independent of insulin sensitivity.

OBJECTIVE: In order to increase significantly the discriminatory potential of Y-STR systems available to the forensic community, we have developed a system capable of simultaneously amplifying 9 Y-STR loci by fluorescence-labeled multiplex PCR technique. METHODS: Primers of STR loci DYS434, GATA-A10, DYS438 and DYS439 were labeled with 6-FAM, primers of STR loci DYS531, DYS557, DYS448 were labeled with HEX, and primers of STR loci DYS456, DYS444 were labeled with TAMRA, respectively. PCR products were analyzed using capillary electrophoresis and GeneScan Software on the ABI Prism310 DNA Analyzer. Series experiments were carried out to evaluate the useful value in forensic application such as the sensitivity, male specificity and genotyping DNA different tissues of the same individual. RESULTS: 9 Y-STR loci were exactly determined following optimization of the polymerase chain reaction. In a sample of 120 males, a total of 105 different haplotypes was identified, 97 of them being unique. Overall, haplotype diversity was 0.996 8. It was proved that genotyping of these 9 Y-STR loci in some sexual crime should be prior to that of automosomal STR. CONCLUSION The results suggest that the newly constructed 9-plex will be very powerful for establishing Y-STR database, population genetic studies and mixture stains identification.
Alleles ; Blood Stains ; Chromosomes, Human, Y/genetics* ; DNA Primers ; Female ; Fluorescence ; Gene Frequency ; Genetics, Population ; Genotype ; Haplotypes ; Humans ; Male ; Polymerase Chain Reaction/methods* ; Sensitivity and Specificity ; Tandem Repeat Sequences

The prokaryotic expression plasmid pQE30-HN of hemagglutinin-neuraminidase (HN) protein gene of bovine parainfluenza virus type 3 (BPIV3) strain HJ-1 was expressed by IPTG induction in E. coli XL1Blue. The recombinant HN protein(rHN) was purified by electroeluting method, and used as coated antigen. An indirect enzyme-linked immunosorbent assay (ELISA) was developed to detect the antibody valence of BPIV3. The best working conditions of ELISA were as follows: the antigen concentration was 6 microg/mL; the serum dilution was 1:50; the blocking reagent was 5% skimmed milk; the blocking time was 60 min at 37 degrees C; the second antibody concentration was 1:10 000; The cut-off value was 0.30. The method revealed a good specificity, no cross-reaction to the positive sera of BCV, IBRV or BRSV was observed. We applied the method to detect 323 serum samples of dairy cow in Heilongjiang Province, the seropositivity rate of BPIV3 was about 58%. The indirect ELISA established provided a technological basis for the development of ELISA kit.
Animals ; Antibodies, Viral ; blood ; Blotting, Western ; Cattle ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; genetics ; Female ; HN Protein ; genetics ; Parainfluenza Virus 3, Bovine ; genetics ; immunology

OBJECTIVEThe pathophysiology of beta-thalassemia is the imbalance of the alpha and non-alpha globin chain which leads to a series of clinical symptoms of hemolytic anemia. Scientists continuously try to explore gene-activated drugs to increase the level of non-alpha globin chain or decrease the level of alpha globin chain in the treatment of beta-thalassemia. To probe into the effects on globin-gene expression of meisoindigo (Me) in cultured erythroid cells derived from peripheral blood, so as to provide the theoretical basis for applying Me in the treatment of beta-thalassemia.

METHODSBy using the two-step liquid culture of erythroid progenitor cells and reverse transcription polymerase chain reaction (RT-PCR), and by using alpha mRNA as an inner control, the level of gamma mRNA and beta mRNA in cultured erythroid cells derived from peripheral blood of 11 patients with severe beta-thalassemia and 6 normal volunteers were measured under the effect of different concentration (2.5 micro mol/L, 5 micro mol/L and 10 micro mol/L) of Me.

RESULTS(1) No statistic significance was found in the ratio of beta/alpha mRNA by Me in cultured cells from both normal individuals and beta-thalassemia. (2) Me can significantly increase the ratio of gamma/alpha mRNA and (beta + gamma)/alpha mRNA (that is non-alpha/alpha mRNA) in cultured cells from normal individuals and beta-thalassemia. The ratio of gamma/alpha mRNA was increased 0.31 - 0.45 times and the ratio of non-alpha mRNA/alpha mRNA increased 0.21 - 0.32 times in Me induced cells from normal individuals. No significant result was observed among the different concentrations of Me (2.5 micro mol/L, 5 micro mol/L and 10 micro mol/L) in normal individuals. With the increasing of Me concentrations, the ratios of gamma/alpha mRNA and alpha/alpha mRNA were increased in cultured cells from beta-thalassemia. The ratio of gamma/alpha mRNA was increased 0.33 - 1.17 times and the ratio of non-alpha/alpha mRNA increased 0.25 - 0.89 times in Me induced cells from beta-thalassemia. There was no significant difference between the concentrations of 2.5 micro mol/L and 5 micro mol/L concentration in beta-thalassemia. However, there was significant difference between the concentrations of 10 micro mol/L and the concentrations of 2.5 micro mol/L and 5 micro mol/L in beta-thalassemia. (3) The increase of the ratio of gamma/alpha mRNA and non-alpha/alpha mRNA in beta-thalassemia was higher than that in normal individual with induction by Me with a higher concentration (10 micro mol/L).

CONCLUSIONMe can raise the ratio of gamma/alpha mRNA and non-alpha/alpha mRNA in cultured erythroid cells derived from peripheral blood of both normal individual and beta-thalassemia in the level of transcription, which can improve the imbalance of the alpha and non-alpha globin chain. So Me has a latent value in the therapy of beta-thalassemia.

Cells, Cultured ; Child ; Child, Preschool ; Erythroid Precursor Cells ; drug effects ; metabolism ; Female ; Gene Expression ; drug effects ; Globins ; genetics ; Humans ; Indoles ; pharmacology ; Infant ; Male ; RNA, Messenger ; drug effects ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction