[Analysis of the results of patch test in 192 patients with hand eczema].

Objective: To investigate the common allergens in the patients with hand eczema. Methods: From November 2014 to March 2016, the patients with hand eczema were tested by the patch test kit of daily life series. Results: The results of the patch test of 192 patients with hand eczema were collected. Allergens were detected in 178 (92.71%) cases. The top 5 allergens were nickel chloride (23.96%) , cobalt chloride (18.75%) , aromatic compounds (17.19%) , nickel sulfate (16.67%) and thimerosal (13.54%). The positive rates of kappa mixture, aromatic compounds, tertiary butyl phenolic resin in males were 16.88% , 14.29% , 11.69% , respectively, which were higher than those (5.22% , 4.35% , 3.48%) in females. Conclusion: Nickel chloride, cobalt chloride, aromatic compounds, nickel sulfate and thimerosal are common allergens in patients with hand eczema.

Immune responses to tyrophagus putrescentiae-induced airway inflammation in mice.

BACKGROUND: Storage mites are a source of aeroallergens that affect patients with allergic rhinitis and asthma. Tyrophagus putrescentiae is a causative factor of airway hypersensitivity, but the mechanisms and pathogenesis of Tputrescentiae-induced allergy are not well understood. OBJECTIVE: This study aimed to develop a murine model of T putrescentiae-induced allergic asthma. METHODS: Immune responses and physiologic variations in immunoglobulins (Ig), leukocyte subpopulations, cytokines, gene expression, pulmonary function, and lung pathology were evaluated after intraperitoneal sensitization and intratracheal challenge with crude extract of T putrescentiae. RESULTS: After sensitization with aluminum hydroxide and challenge with T putrescentiae in mice, levels of T putrescentiae-specific IgE and IgG1 in sera increased significantly compared to the normal saline group (P < .01): Values for inflammatory leukocytes (neutrophils and eosinophils) and cytokines (interleukin [IL] 4, IL-5, and IL-13) increased significantly after sensitization. In terms of pulmonary function, pause values were significantly enhanced in T putrescentiae-sensitized mice after intratracheal challenge with T putrescentiae (P < .05). Expression of type 2 helper T cell (T(H)2)-related genes (IL4, IL5, IL13, and RANTES), T(H)2-specific transcription factor (GATA-3), and proinflammatory genes (IL6), and T(H)(H)17-related genes (IL17F) increased significantly after airway challenge. Sensitization with T putrescentiae crude extract led to inflammation of lung tissue, thickening of the tracheal wall, and tracheal rupture. CONCLUSIONS: Intraperitoneal sensitization followed by intratracheal challenge with crude extract of T putrescentiae can induce airway inflammation in BALB/c mice. The symptoms observed in a mouse model of allergic asthma, in terms of immune and clinical parameters, are reminiscent of the symptoms of allergic asthma in humans. A mouse model can be used to evaluate the therapeutic effectiveness of drugs on T putrescentiae-induced airway inflammation in humans.

Comparison between the radial forearm and groin soft tissue free flaps for reconstruction in patients with oral cavity cancer: a quality of life analysis.

The purpose of this study was to compare the effects of the radial forearm free flap (RFFF) and groin soft tissue free flap (GSFF) on the quality of life (QoL) of patients undergoing reconstructive surgery after resection for oral cancer. A retrospective analysis of 48 patients was performed. The Vancouver Scar Scale (VSS), University of Washington Quality of Life (UW-QOL) questionnaire, and 14-item Oral Health Impact Profile (OHIP-14) questionnaire were used to evaluate the donor site scars and QoL of the patients. The postoperative hospital stay was significantly longer in the RFFF group than in the GSFF group (P = 0.001). Furthermore, the total VSS score (P = 0.011), VSS score for pigmentation (P < 0.001), and OHIP-14 scores for psychological discomfort (P = 0.026) and social disability (P = 0.044) were all significantly higher in the RFFF group than in the GSFF group, while the UW-QOL scores for appearance (P = 0.037) and mood (P = 0.036) were significantly lower in the RFFF group than in the GSFF group. Compared with the RFFF, the GSFF scar is more concealed, with better aesthetics at the donor site, and this flap can result in improved postoperative QoL for patients with oral cancer.

Prognostic impact of anatomical extent of metastatic lymph node on gastric cancer: a propensity score matching study.

PURPOSE: Current gastric cancer staging systems overlook the anatomic extent of metastatic lymph nodes (AEMLNs). This study aimed to analyze the prognostic impact of AEMLNs on gastric cancer (GC). METHODS: GC patients with metastatic lymph nodes (MLNs) undergoing curative surgery were retrospectively reviewed and assigned to perigastric (MLNs in station 1-6, PG) and extraperigastric group (7-12, with or without MLNs in PG area, EPG). Overall survival (OS), disease-free survival (DFS) and recurrence patterns were compared before and after 1:1 propensity score matching (PSM). RESULTS: 662 patients were enrolled, 341 (51.5%) and 321 (48.5%) of whom were in the PG and EPG, respectively. After PSM (n = 195), EPG showed poorer 5-year OS (43.4% vs 54.5%, p = 0.014) and DFS (65.0% vs 73.4%, p = 0.068) than PG. EPG had higher incidence of peritoneal recurrence (PR) than PG (19.4% vs 7.4%, p = 0.002). Multivariate analysis identified AEMLNs as prognostic factor for OS [HR = 1.409, 95% confidence interval (CI) 1.062-1.868), DFS (HR = 1.600, 95% CI 1.059-2.416) and PR (HR = 3.708, 95% CI 1.685-8.160). CONCLUSIONS: The anatomic extent of metastatic lymph nodes has an independent prognostic role for GC. Including this element may improve the accuracy of current staging systems.

[Role of Testin in nasopharyngeal squamous cancer with high ability of metastasis].

Objective:To explore the influence and regulatory mechanism of TES gene on proliferation and migration of nasopharyngeal squamous cancer(NSPC) 5-8F cell.Method:DNA fragment encoding TES was obtained by RT-PCR method from the human highly metastatic nasopharyngeal squamous carcinoma cell line 5-8F. we identified the recombinant plasmid pEGFP-N1-TES by RT-PCR and DAN sequencing. we stablely transfected the pEGFP-N1-TES into the human highly metastatic nasopharyngeal squamous carcinoma cell line 5-8F, and detected the expression of TES by the RT-PCR and Western-blot method. And detected the impact of 5-8F cells transfection by flow cytometry and scratch tests.Result:Flow cytometry analysis showed that the apoptotic in 5-8F/pEGFP- N1-TES was significantly higher than non-transected TES and 5-8F/pEGFP-N1,and the differences were statistically significant(P<0.05).Cell scratch experiments showed that the 5-8F/pEGFP-N1-TES group cell migration rate was obviously lower than nontransected TES and 5-8F/pEGFP-N1 group in the first 12 h, 24 h and 48 h.The difference was significant(P<0.01).Conclusion:The stable transfectant cell model was established successfully. TES in vitro could significantly increase apoptosis and reduce the athletic ability. And thus TES gene might be a novel candidate of tumor-suppressor.

[Targeted degradation of epidermal growth factor receptor in nasopharyngeal carcinoma by chimeric molecule].

Objective:The aim of this study was to investigate targeted degradation of the epidermal growth factor receptor (EGFR) by chimeric molecules (EGF-PROTAC) via the ubiquitin-proteasome pathway on human nasopharyngeal carcinoma CNE-2 cells and demonstrate the regulative effect on the proliferation and apoptosis of the CNE-2 cells.Method:After the EGF-PROTAC treating CNE-2 cells in vitro, the biological effects of the EGF-PROTAC was detected by western blot, CKK-8 assay, flow cytometry and Transwell migration assay in CNE-2 cells.Result:The expression level of EGFR proteins in the EGF-PROTAC treated group was lower than the control group (P< 0.05); CKK-8 assay results showed that CNE-2 cells survival rate at 3, 6, 9 and 12h decreased greatly than the control group (P< 0.05); Flow cytometry indicated that the apoptosis index of the CNE-2 cells in EGF-PROTAC treated group was significantly higher than the control group (P< 0.05); The invasion ability detected that the number of CNE-2 cells in the EGF-PROTAC treated group was significantly lower than the control group (P< 0.05).Conclusion:The chimeric molecule (EGF-PROTAC) can target the degradation of epidermal growth factor receptors (EGFR) and effectively inhibit the growth of the CNE-2 cells and promote apoptosis in vitro.

[Effect of ShRNA targeting silencing of VTCN1 gene on nasopharyngeal carcinoma cells].

Objective:The aim of this study is to investigate the expression of VTCN1 in nasopharyngeal carcinoma cells and targeting silencing of VTCN1 gene by infected with lentiviral vector to inhibit the proliferation and invasion in nasopharyngeal carcinoma cells. Method:The VTCN1 expression level of different nasopharyngeal carcinoma cells was detected by RT-PCR and Western blotting, the cell lines of the most expression level were seleted to conduct the Subsequent experiments; The lentiviral vector of silenced VTCN1 was transfected into HNE2 cells with VTCN1 expression by lipofectamine 2000, and stable cell lines were screened. Then, the silencing efficiency was detect by RT-PCR and Western blotting; The proliferation and invasion abilities of nasopharyngeal carcinoma cells after VTCN1 gene silencing were detected by ckk-8 and Transwell invasion assay, respectively. The phosphorylation levels of JAK/STAT proteins in nasopharyngeal carcinoma cells with VTCN1 gene silencing were detected by Western blotting. Result:RT-PCR and Western blotting detected that stable transfection of VTCN1ShRNA into HNE2 cells resulted significantly declined expression of VTCN1 (P<0.05); The proliferation and invasion abilities of HNE2 cells were significantly decreased (P<0.05) and phosphorylation level of the JAK, STAT proteins were significantly decreased (P<0.05). Conclusion:VTCN1 ShRNA can effectively silence the expression of VTCN1, and significantly inhibits the proliferation and invasion of HNE2 cells. It may be related to down regulation of protein activity in JAK/STAT signaling pathway..

[Expression and clinical significance of Testin in nasopharyngeal carcinoma].

Objective:Our purpose was to investigate the expression of Testin gene,and its possible relationship with the clinicopathological features of human nasopharyngeal carcinoma.Method:The expression of Testin in nasopharyngeal carcinoma tissues were detected by immunohistochemistry methods,semi-quantitative reverse transcriptase polymerase chain reaction and Western blot.The correlations of Testin to clinicopathologic features of nasopharyngeal carcinoma were analyzed.Result:The mRNA level of Testin was down-regulated in human nasopharyngeal carcinoma.The positive rate of Testin protein was significantly lower in human nasopharyngeal carcinoma tissues than that in nomal tissues;The protein level of Testin was down-regulated in cancers as compared with corresponding normal tissues.Testin expression was positively correlated with the differentiation of nasopharyngeal carcinoma.Meanwhile,differences in gender and age were not significance(P>0.05 respectively) .There was a significant correlation between invasion,distant metastasis and differentiation degree and Testin expression(P<0.05 respectively).Conclusion:The decreased expression of Testin gene may play an importmant role in the development of esophageal squamous cancer.Thus Testin gene might be a novel candidate of tumor-suppressor.It may be an objective marker for prognostic factor and malignant level for nasopharyngeal carcinoma.

Caspase 3, periodically expressed and activated at G2/M transition, is required for nocodazole-induced mitotic checkpoint.

Caspases have been known for several years for their involvement in executing apoptosis, where unwanted or damaged cells are eliminated. Surprisingly, after analysis of the relevant data set from the Stanford microarray database, we noticed that the gene expression pattern for caspase 3, but not for caspase 1, 6, 7, 8, 9, or 10, undergoes periodic change in the HeLa cell cycle. In this study, we have demonstrated that caspase 3, but not other caspases, is upregulated and activated just prior to mitosis. Pretreatment of human hepatoma cells with a caspase 3 inhibitor z-DEVD-FMK, prior to the treatment with an antimicrotubule drug nocodazole, abrogates the mitotic arrest, suggesting that caspase 3 (or a caspase 3-like enzyme) might be involved in mitotic-spindle checkpoint. The studies not only characterize caspase 3 as a cell cycle-regulated protein, but also link the protein to nocodazole-dependent mitotic checkpoint, greatly expanding the understanding of caspase 3.

Isolation and in vitro translation of messenger RNA from the American cockroach.

Total RNA was extracted from the whole body of American cockroach (Periplaneta americana) using chaotropic salt guanidine isothiocyanate in the presence of 2-mercaptoethanol (2-ME). Polyadenylated mRNA was isolated by oligothymidylic acid-cellulose chromatography and mRNA was translated using a rabbit reticulocyte lysate system. The translation, as judged by the incorporation of 35S-methionine, was obtained with poly(A)+ RNA, where an approximately 9.5-fold increase in label incorporation over control was achieved. Analysis of translation products by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) in combination with autoradiography showed that many proteins with apparent molecular weights ranging from 12 to 200 kD were synthesized, and no labelled proteins were found with negative RNA control and poly(A)- RNA. Immunoprecipitation studies performed using polyclonal and monoclonal antibodies revealed that synthesized proteins of MW 90, 78, 72, 49, 45, and 26 kD corresponded with previously identified principal and major allergens of American cockroach from our laboratory. In addition, the allergenicity of the translation mixtures was also confirmed by fluoroallergosorbent test (FAST) inhibition studies with IgE antibodies of human reaginic serum pool.

Involvement of cyclin-dependent kinase activities in CD437-induced apoptosis.

A novel synthetic retinoid, 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid (CD437), is a selective ligand of the RARgamma nuclear receptor. We examined the in vitro effects of CD437 and found that CD437 induces S phase arrest within 24 to 48 h, followed by cell death, in the p53-negative Hep3B and the p53-positive HepG2 human hepatoma cell lines. Based on observations of cellular and nuclear fragmentation, chromatin condensation, and DNA fragmentation, the CD437-mediated cell-killing effect appears to be due to apoptosis. On morphological examination, a number of CD437-treated cells were found to have increased 5- to 10-fold in size and persisted as single giant cells without cell division, while the remainder underwent nuclear division (multiple nuclei) but were unable to complete cytokinesis, and finally all died by apoptosis. In HepG2 cells that possessed wild-type p53, CD437-induced S phase arrest and apoptosis were accompanied by the up-regulation of cyclin A, cyclin B, p53, p21(CIP1/Waf1), Bad, and Bcl-Xs proteins and by a decrease in Bcl-2 protein levels. In Hep3B cells, CD437-mediated S phase arrest and apoptosis were also associated with a concomitant up-regulation of cyclin A, cyclin B, Bad, and Bcl-Xs. However, Hep3B cells did not express p53 or Bcl-2 messages. Olomoucine and roscovitine, the potent p34(cdc2) and CDK2 inhibitors, effectively blocked CD437-mediated cyclin A- and B-dependent kinase activation and prevented CD437-induced cell death. Furthermore, antisense oligonucleotide complementary to cyclin A and B mRNA significantly rescued CD437-induced apoptosis. These findings suggest that activation of cyclin A- and B-dependent kinases is a critical determinant of apoptotic death mediated by CD437.

Differential effects of phorbol ester on growth and protein kinase C isoenzyme regulation in human hepatoma Hep3B cells.

PMA has both mitogenic and antiproliferative effects on human hepatoma Hep3B cells. In response to low PMA concentration (10 nM), Hep3B cells displayed an increasing proliferation potentiation. At high PMA concentration (1 microM) Hep3B cells exhibited modest cytostatic effects. Determinations of protein kinase C (PKC) activity in PMA-treated cells revealed that alterations in PKC activity are associated with proliferative capacity. The decrease in PKC activity mediated by a high dose of PMA was accompanied by cell growth inhibition. Increases in PKC activity mediated by a low dose of PMA were consistent with proliferation stimulation. Immunoblot analysis showed that there are at least six PKC isoenzymes: alpha, delta, epsilon, mu, zeta and iota/lambda, constitutively expressed in Hep3B cells. Cellular fractionation and immunocytochemical staining results demonstrated that both 10 nM and 1 microM PMA treatments induced a marked translocation of PKC-alpha from cytosol to membrane or nuclear fraction within 5-30 min. At the same time PKC-delta and epsilon were translocated from the membrane to nuclear fraction. In addition, prolonged treatment with 1 microM PMA, but not with 10 nM PMA, selectively mediated the down-regulation of these three PKC isoenzymes. The distinct effects of different concentrations of PMA on cell proliferation and PKC-alpha, delta and epsilon isoenzyme modulation support the involvement of these three PKC isotypes in the mechanism of action of Hep3B cells in cell growth events.

Electrochemical property: Structure relationships in monoclinic Li(3-y)V2(PO4)3.

Monoclinic lithium vanadium phosphate, alpha-Li(3)V(2)(PO(4))(3), is a highly promising material proposed as a cathode for lithium-ion batteries. It possesses both good ion mobility and high lithium capacity because of its ability to reversibly extract all three lithium ions from the lattice. Here, using a combination of neutron diffraction and (7)Li MAS NMR studies, we are able to correlate the structural features in the series of single-phase materials Li(3-y)V(2)(PO(4))(3) with the electrochemical voltage-composition profile. A combination of charge ordering on the vanadium sites and lithium ordering/disordering among lattice sites is responsible for the features in the electrochemical curve, including the observed hysteresis. Importantly, this work highlights the importance of ion-ion interactions in determining phase transitions in these materials.

Comparison of polymerase chain reaction, monoclonal antibody based enzyme immunoassay, and cell culture for detection of Chlamydia trachomatis in genital specimens.

An enzyme immunoassay (EIA; AntigEnz Chlamydia; Northumbria Biologicals, Northumberland, United Kingdom) and a polymerase chain reaction (PCR) assay (Genemed Biotechnologies, San Francisco, CA) were evaluated for the detection of Chlamydia trachomatis in urogenital specimens. Of 324 specimens, 23 were positive by cycloheximide-treated McCoy's cell culture method. Of 23 culture-positive specimens, 20 and 22 were found to be positive by EIA and PCR, respectively. Among 301 culture-negative samples, 4 were found to be positive by both PCR and EIA, 2 were PCR-positive and EIA-negative, and 3 PCR-negative and culture-negative specimens were found to be positive by EIA. In comparing nonculture methods with cell culture technique, combined sensitivities of 87.0% (90.9% in men and 83.3% in women) and 95.6% (90.9% in men and 100% in women), and specificities of 97.7% (99.4% in men and 95.7% in women) and 98.0% (99.4% in men and 96.5% in women) are achieved by EIA and PCR, respectively. The PCR proved to be the more sensitive of these two nonculture methods, and it can be used for the rapid diagnosis of C. trachomatis urogenital infection.

PIP: A total of 324 endocervical or urethral swab specimens were collected from patients attending the sexually transmitted diseases clinic in China Medical College and Taichung Veterans General Hospital, Taichung, Taiwan. A total of 324 urethral (from 171 men) and endocervical (from 153 women) specimens were examined by cell culture, AntigEnz Chlamydia enzyme immunoassay (EIA) (Northumbria Biologicals, Northumberland, United Kingdom), and polymerase chain reaction (PCR) for the presence of Chlamydia trachomatis. Of 324 specimens, 23 (11 from men and 12 from women) were found to be positive by the McCoy's cell culture method. Of the 23 culture-positive samples, 20 (10 from men and 10 from women) were found to be positive by both EIA and PCR, 2 (both from women) were PCR-positive and EIA-negative, and the other (from a male patient) was found to be negative by EIA and PCR. Of 301 culture-negative specimens, 292 were found to be negative by both PCR and EIA. Of the remaining specimens, 4 (1 from a male patient and 3 from female patients) culture-negative and EIA-positive and 2 (both from women) culture-negative and EIA-negative specimens were found to be positive by PCR. In addition, 3 (all from women) PCR-negative and culture-negative specimens were found to be positive by EIA. If a positive cell culture result is considered as the standard, PCR demonstrated a sensitivity of 90.9% (10/11) in men and 100% (12/12) in women, and EIA revealed a sensitivity of 90.9% (10/11) in men and 83.3% (10/12) in women. Specificities of 99.4% (159/160) in men and 96.5% (136/141) in women, and of 99.4% (159/160) in men and 95.7% (135/141) in women were obtained by PCR and EIA, respectively. Combining the data for men and women, sensitivities of 95.6% (22/23) and 87.0% (20/23), and specificities of 98.0% (295/301) and 97.7% (294/301) were achieved by PCR and EIA, respectively. The PCR proved to be the more sensitive of these two nonculture methods, and it can be used for the rapid diagnosis of C. trachomatis urogenital infection.

Induction of p21(CIP1/Waf1) and activation of p34(cdc2) involved in retinoic acid-induced apoptosis in human hepatoma Hep3B cells.

The biological activity of retinoic acid (RA) was examined in human hepatoma Hep3B cells. Under serum-deprived conditions, RA induced S/M-phase elevation and mitotic index increase within 24 h, followed by apoptosis. This RA-induced apoptosis was accompanied by p53-independent up-regulation of endogenous p21(CIPI/Waf1) and Bax proteins, as well as activation of p34(cdc2) kinase, and increase of Rb2 protein level and phosphorylation pattern. In addition, RA had no effect on the levels of Bcl-XL; Bcl-XS; cyclins A, B, D1, D3, or E; or Rb1 expression but markedly down-modulated Cdk2 kinase activity and reduced Cdk4 expression. RA also slightly delayed p27(Kip1) expression. Olomoucine, a potent p34(cdc2) and Cdk2 inhibitor, effectively blocked RA-mediated p34(cdc2) kinase activation and prevented RA-induced apoptosis. Furthermore, antisense oligonucleotide complementary to p21(CIP2/Waf1) and p34(cdc2) mRNA significantly rescued RA-induced apoptosis. Our data indicate that p21(CIP2/Waf1) overexpression may not be the only regulatory factor necessary for RA-induced apoptosis in human hepatoma Hep3B cells. RA treatment leads to Rb2 hyperphosphorylation, and p34(cdc2) kinase activation is coincident with an aberrant mitotic progression, followed by appearance of abnormal nucleus. This aberrant cell cycle progression appeared requisite for RA-induced cell death. These findings suggest that inappropriate regulation of the cell cycle regulators p21(CIP2/Waf1) and p34(cdc2) is coupled with induction of Bax and involved in cell death with apoptosis when Hep3B cells are exposed to RA.