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 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 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 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

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

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

OBJECTIVETo observe and compare the medium-long-term efficacy of Kurokawa's and modified Kurokawa's double door laminoplasty for the treatment of cervical disorders.

METHODSA retrospective analysis was performed to compare the outcomes and complications between two kinds of operations on 172 cases from January 2002 to December 2010, including 106 cases of cervical spondylotic myelopathy, 52 cases of cervical stenosis, 21 cases of cervical ossification of the posterior longitudinal ligament. Patients were divided into two groups according to two surgical methods: traditional group, including 51 male and 18 female patients, with mean age of (56 ± 18) years (35-76 years); modified group, including 75 male and 28 female patients, with mean age of (58 ± 20)years (35-80 years). The two groups were comparable and compared according to different data using t test, χ(2) test and rank sum test.

RESULTSAll patients were followed up continuously for (52 ± 33)months, 123 patients were followed up ≥ 2 years, 71 patients ≥ 5 years. All patients' Japanese Orthopaedic Association (JOA) score improved significantly at the latest follow-up(t = 3.420, P < 0.01); no significant difference between the patients' JOA score improvement rate of two groups. The postoperative incidence rate of axial symptoms in patients of modified group (3.9%) was significantly lower than the traditional group (14.5%) (χ(2) = 7.548, P < 0.05), and cervical intervertebral activity decreased in the modified group was better than the traditional group in the first 3 months postoperatively (27% ± 6% vs. 19% ± 4%,Z = 6.34, P < 0.05), but during the medium-long-term follow-up, no significant difference in the cervical intervertebral activity decreased between two groups.

CONCLUSIONSMedium-long-term efficacy of Kurokawa's and modified Kurokawa's double door laminoplasty is satisfied and reliable. Avoiding damaging of semispinalis cervicis insertion in spinous process of C2, the modified operation method can protect the extensor group of the neck muscle and reduce the incidence of postoperative axial symptoms better.

Adult ; Aged ; Aged, 80 and over ; Cervical Vertebrae ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neurosurgical Procedures ; methods ; Retrospective Studies ; Spinal Osteophytosis ; surgery ; Treatment Outcome

OBJECTIVETo investigate the loss of heterozygosity (LOH) on chromosomal arms 13q and 14q in nasopharyngeal carcinoma (NPC) using 21 microsatellite polymorphic markers and to study whether there is a correlation between LOH and clinicopathologic parameters and/or Epstein-Barr virus (EBV) infection in NPC.

METHODSSixty cases of NPC were studied using polymerase chain reaction based microsatellite analysis with genescan and genotyping techniques.

RESULTSLOH was detected on 13q in 78% of NPC tumors, high frequency LOH loci (more than 30%) clustered to 13q12.3-q14.3 and 13q32. On chromosome 14q, LOH was detected in 80% of NPC tumors; high frequency LOH loci clustered to 14q11-q13, 14q21-q24 and 14q32. High frequency LOH at 13q31-q32 correlated with a lower level of EBV infection; LOH on chromosome 14q was closely associated with poor differentiation of NPC tumor cells.

CONCLUSIONOur results suggest that in NPC, LOH on chromosome 13q and 14q are common genetic events, and putative tumor suppressor genes (TSG) residing in these regions may be involved in tumorigenesis.

Adult ; Aged ; Chromosomes, Human, Pair 13 ; genetics ; Chromosomes, Human, Pair 14 ; genetics ; DNA, Neoplasm ; genetics ; Female ; Gene Frequency ; Humans ; Loss of Heterozygosity ; Male ; Microsatellite Repeats ; Middle Aged ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Statistics as Topic

OBJECTIVETo investigate the outcome of surgical reconstruction of the tongue after hemiglossectomy with reinnervated rectus abdominis musculoperitoneal flaps in the treatment of tongue cancer.

METHODSFive patients underwent immediate reconstruction of the tongue and oral floor defects with rectus abdominis musculoperitoneal flaps after resection of squamous cell carcinoma of tongue. The rectus abdominis musculoperitoneal flap consists of the rectus muscle, posterior rectus sheath, peritoneum, the 10 th, 11th, 12th intercostal nerves and the vascular pedicle that includes the deep inferior epigastric artery and veins. During the operation a reinnervated rectus abdominis musculoperitoneal free flap, in which the intercostal nerves were anastomosed to the descending branch of hypoglossal nerve, was grafted to remaining tongue stump.

RESULTSAll patients recovered uneventfully from surgery, with no immediate postoperative complications. All transplanted flaps survived. The peritoneum was replaced by squamous epithelium 8 weeks after surgery. The average follow-up period was 10 months. During the follow-up period the contour of the reconstructed tongues was satisfactory. The patients demonstrated good functional mobility of the reconstructed and remaining tongue. The swallowing and speech function was nearly at normal levels and the patients could ingest a solid or semisolid diet.

CONCLUSIONSReconstruction of the tongue with rectus abdominis musculoperitoneal flaps after hemiglossectomy is a suitable, cosmetically acceptable method. Long-term follow-up is needed for reaching some final conclusions.

Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Peritoneum ; transplantation ; Reconstructive Surgical Procedures ; methods ; Rectus Abdominis ; transplantation ; Surgical Flaps ; Tongue ; surgery ; Tongue Neoplasms ; surgery ; Treatment Outcome

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