OBJECTIVE: To express the recombinant SPLUNC1 protein in HNE1 cells and to study its function of bactericidal and binding to lipopolysaccharide (LPS). METHODS: Full length of SPLUNC1 gene was cloned into pCMV-tag4A vector and stably transfected into HNE1 cell lines, the supernatant of cell cultures was collected. After being treated with the supernatant, the Pseudomonas aeruginosa was seeded to LB soft agar plate, and the bacteria clones were counted and analyzed. For in vitro LPS binding assay, LPS was coated to 96-well plates. We incubated in the plate with SPLUNC1 protein, and detected the binded SPLUNC1 protein by ELISA. Incubating the FITC-LPS with the SPLUNC1 stably transfected or control cells, the intracellular intensity of fluorescence was observed under the fluorescence microscope. RESULTS: SPLUNC1 inhibited the bacteria clone formation obviously. Although the binding efficiency of LPS and SPLUNC1 in vitro was very low, more FITC-LPS entered into the SPLUNC1 stably transfected cells. CONCLUSION SPLUNC1 can inhibit the growth of Pseudomonas aeruginosa and bind LPS, and play an important defensive role in innate immunity of the upper airway.
Cell Line, Tumor ; Glycoproteins ; isolation & purification ; pharmacology ; Humans ; Membrane Proteins ; chemistry ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Phosphoproteins ; isolation & purification ; pharmacology ; Pseudomonas aeruginosa ; drug effects ; Respiratory Mucosa ; chemistry ; immunology ; Respiratory System ; chemistry ; immunology ; Transfection

OBJECTIVETo analyze the effect of gold standard, blind comparison and different cut-points choosing on screening techniques assessment, and to promote the application of evidence-based medicine theory in screening study.

METHODSA screening study for cervical cancer in rural China in 1999, where 1997 women had been tested for pathology as gold standard and simulating situations without gold standard, blind comparison and under different cut-points. Indices such as detectable rate, sensitivity and specificity were calculated for each technique. Receiver operating characteristics (ROC) curves were drawn and areas under ROC curves between screening techniques were tested.

RESULTSWithout gold standard, diagnostic techniques could not be evaluated correctly, and without the blind comparison, the sensitivity and specificity of the tests would be subjectively increased. Furthermore, use of different cut-points led to different sensitivities and specificities of test.

CONCLUSIONSGold standard, blind comparison and perfect cut-points can improve the quality of screening test and drawing ROC curves is an effective way to confirm cut-points and evaluate diagnostic techniques. It is necessary to enforce the application of evidence-based medicine theory in scientific research.

Adult ; Clinical Laboratory Techniques ; methods ; Evaluation Studies as Topic ; Evidence-Based Medicine ; Female ; Humans ; Mass Screening ; Middle Aged ; ROC Curve ; Technology Assessment, Biomedical ; methods ; Uterine Cervical Neoplasms ; prevention & control

OBJECTIVETo determine the association between viral load of high risk type human papillomavirus (HR-HPV) and stage of cervical intraepithelial neoplasia (CIN) lesion.

METHODSCervical exfoliated cells were collected from 1997 women aged 35-45 in a cross-sectional screening study. HPV DNA was detected by hybrid capture 2 (HC2) system, and viral load was measured by the ratios of relative light units compared to standard positive control (RLU/PC). Log10RLU/PC were categorized into four groups: negative (< 0), low viral load (0 - 1.12), medium viral load (1.13 - 2.23), and high viral load (2.24 - 3.37). Cervical lesions were diagnosed by biopsies as normal, CIN 1, CIN 2-3, and squamous cervical cancer (SCC). Association between HR-HPV and CINs were evaluated by unconditional multinomial logistic regression.

RESULTS100% (12/12) SCC, 97.3% (72/74) of CIN 2-3, 58.3% (74/127) of CIN 1, and 11.5% (205/1784) of normal women were positive for HPV DNA. The median log10RLUs for the positive women with SCC, CIN 2-3, CIN 1 and in normal women were 2.60, 2.32, 2.18 and 1.18 respectively. The odds ratio (OR) between low viral load of HPV DNA and CIN 1 was 3.8 (1.9 - 7.3) while between high viral load and CIN 2-3 was OR=865.9 (200.1 - 3738.0) which showed that higher viral load could increase the risk of cervical lesions (P <0.001).

CONCLUSIONBoth cervical cancer and CINs were highly influenced by HR-HPV viral load.

Adult ; Cervical Intraepithelial Neoplasia ; epidemiology ; virology ; China ; epidemiology ; Cross-Sectional Studies ; DNA, Viral ; analysis ; Female ; Humans ; Logistic Models ; Mass Screening ; Middle Aged ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus Infections ; epidemiology ; virology ; Risk Factors ; Uterine Cervical Neoplasms ; epidemiology ; virology ; Viral Load

OBJECTIVESTo investigate the prevalence of oncogenic type of human papillomavirus (HPV) infection and identify the high risk population for conducting immuno/chemoprevention of cervical cancer.

METHODSAll married women aged 30 to 50 with no history of hysterectomy, pelvic radiation and non-pregnant from certain villages of Xiangyuan and Yangcheng County were invited. This study was conducted through two phases. In phase one, subjects sampled the vaginal secretions using the collectors after signing the informed consent. And physicians sampled exfoliated cells from cervix in the phase two. All the specimens were tested with the Hybrid Capture 2 test. The data was managed and analyzed by VFP and SPSS software.

RESULTSThere were 9,683 women participated in this study. Local women welcomed this study and population compliance rate was 75.4%. In tested population, we found 2,666 subjects of HPV DNA positive and HPV prevalence was 27.5%. The rates of different age group were 24.5% (30-34 yrs), 27.4% (35-39 yrs), 28.2% (40-44 yrs), 27.4% (45-50 yrs) respectively and had no significant differences (P = 0.604). The rates were slightly increased with the higher education level and had no differences (P = 0.106). The rate in mountain areas was higher than that in half-mountain areas (P = 0.001).

CONCLUSIONSThe prevalence of HPV infection is indeed high in this region. Local women and health professionals welcome the activities of cervical cancer screening and prevention. It is an emergent task to improve their sanitary condition and prevent them from cervical cancer in these women. A women health cohort is established successfully among high HPV exposed women in rural China. The extensive biologic specimen repository has been successfully established to simultaneously study the etiology, early detection, and immuno/chemoprevention of cervical cancer.

Adult ; China ; epidemiology ; DNA, Viral ; analysis ; Female ; Humans ; Mass Screening ; Middle Aged ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; epidemiology ; prevention & control ; Prevalence ; Tumor Virus Infections ; epidemiology ; prevention & control ; Uterine Cervical Neoplasms ; epidemiology ; prevention & control ; virology ; Vaginal Smears

OBJECTIVETo get the genotype and allele frequency distributions of 8 short tandem repeat (STR) loci on chromosome 3p (D3S1297, D3S1489, D3S1266, D3S1568, D3S1289, D3S1300, D3S1285 and D3S3681) in Chinese Han population in Hunan area.

METHODSBlood samples were collected from the random Han individuals in Hunan and the whole genomic DNA was extracted. STR loci were amplified by multiplex-PCR technique and genotyped by ABI 377 sequencer.

RESULTSNinety-one alleles were detected, with frequencies ranging from 0.002 to 0.431, and these alleles constituted 312 genotypes. All the 8 loci met Hardy-Weinberg equilibrium. The statistical analysis of 8 STR loci showed the heterozygosity (H) >or= 0.729, the discrimination power (DP) >or= 0.725, the probabilities of paternity exclusion (PPE) >or= 0.596, and the polymorphic information content (PIC >or= 0.682). The result indicated that there was a significant difference between Han ethnic group and the white and the black.

CONCLUSIONThese results could serve as valuable data to enrich the Chinese genetic database and play an important role in Chinese population genetic and forensic medical application.

Adult ; Alleles ; China ; Chromosomes, Human, Pair 3 ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Microsatellite Repeats ; genetics ; Polymorphism, Genetic

OBJECTIVETo investigate the relationship of nasopharyngeal carcinoma (NPC) with the high frequency allele imbalance locus D6S1581, and the NPC associated gene FBXO30 which is located near D6S1581.

METHODSGenescan was used to genotype D6S1581 of 12 NPC pedigrees, 85 sporadic NPC patients and 181 normal volunteers. Then parametric/nonparametric linkage analysis and association analysis were performed.

RESULTSD6S1581 was linked with NPC, a Lod score of 2.611436 (P=0.00245) was obtained, and a significant difference in allele frequency was observed between familial NPC and control (P<0.005).

CONCLUSIONThese results suggest that D6S1581 is highly associated with NPC, and there may be one or more NPC associated genes near D6S1581, including FBXO30.

Adult ; Aged ; Aged, 80 and over ; Alleles ; China ; F-Box Proteins ; genetics ; Female ; Gene Frequency ; Genetic Linkage ; Genetic Predisposition to Disease ; genetics ; Humans ; Male ; Microsatellite Repeats ; genetics ; Middle Aged ; Nasopharyngeal Neoplasms ; genetics ; Pedigree