Objective:To determine the expression of the full-length (NK1R-FL) and truncated (NK1R-Tr) neurokinin-1 receptor (NK1R) and the neurokinin-2 receptor (NK2R) in breast cancer tissues and cell lines, as well as to study the effects of the NK1R and NK2R antagonists on the growth of breast cancer cells. Methods:Immunohistochemistry and Western blot assays were used to detect NK1R, NK1R-FL, and NK2R expression in clinical samples of primary breast cancer tissue, benign lesions, and normal breast tissue, as well as in different breast cancer cell lines. Cell proliferation and soft agar growth tests were performed on cells treated with the NK1R and NK2R antagonists to study the ectopic overexpression of NK1R-FL and NK1R-Tr in breast cancer cell lines. Results:Total NK1R expression was detected in the breast cancer tissues, benign lesions, and normal breast tissues. Compared with the normal breast epithe-lia and benign breast lesions, the expression levels of NK1R-FL and NK2R decreased in the carcinoma. These changes were also relat-ed to the carcinoma type, histological grade, lymph node metastasis, HER2 and Ki-67 expression, and estrogen and progesterone recep-tors in breast cancer. The expression levels of NK1R-FL and NK2R were high in the HBL-100 breast cell lines of para-neoplastic tis-sues, but NK1R-Tr expression was low. The MDA-MB-231, T-47D, and MCF-7 cells only expressed NK1R-Tr. NK1R-Tr or NK1R-FL overexpression caused the decreased inhibition rate or increased levels of the NK1R and NK2R antagonists in the breast cancer cells. Conclusion:NK1R-FL and NK2R are co-expressed in normal cells. NK1R-Tr is highly expressed in breast cancer cells and exerts nega-tive feedback to regulate NK1R-FL and NK2R expression in all cells, especially cancer cells.

Adult ; Antigens, Neoplasm ; metabolism ; Gastrointestinal Neoplasms ; metabolism ; pathology ; surgery ; Gastrointestinal Stromal Tumors ; pathology ; Humans ; MART-1 Antigen ; Male ; Neoplasm Proteins ; metabolism ; Osteoclasts ; pathology ; S100 Proteins ; metabolism ; Sarcoma, Clear Cell ; metabolism ; pathology ; surgery ; Soft Tissue Neoplasms ; metabolism ; pathology ; surgery

Objective To study the changes of antigen-specific T helper type 1 (Th1) cells and Th2 cells in patients with pemphigus vulgaris (PV) at different stages,so as to better understand the roles of autoreactive T cells in PV.Methods The DG3 (96-112) peptide was synthesized.Twenty-four patients with PV and 10 health checkup examinees were included in this study.Peripheral blood mononuclear cells (PBMCs) were obtained from the health checkup examinees and all the patients before treatment and seven patients about one month after treatment,and stimulated with the DG3 peptide of 10 mg/L for different durations.Then,enzyme-linked immunospot (ELISPOT) assay was performed to count the number of DG3-specific IFN-γ+ Th1 cells,IL-4+ Th2 cells and memory B cells.One-way analysis of variance (ANOVA) and t test were done to compare the number of cells between different groups,and Pearson correlation coefficient was used to evaluate the correlation among Th cells,memory B cells and anti-desmoglein 3 (Dsg3) antibody titers.Results In these patients,the male to female ratio was 1.67 ∶ 1,and the average age was (56.59 ± 14.66) years.Compared with the health check-up examinees,the patients with PV showed a higher absolute number of DG3-specific IFN-γ+ Th1 cells (420.18 ± 350.29 vs.145.12 ± 86.56,t =3.25,P ＜ 0.05),but a similar absolute number of specific IL-4+ Th2 cells (366.76 ± 192.44 vs.335.88 ± 164.96,P＞ 0.05) per 5 × 105 PBMCs.The percentage of DG3-specific IL-4+ Th2 cells in Th2 cells was 37.03％ ± 23.44％,and the percentage of IFN-γ+ Th1 cells was 23.62％ ± 16.77％ in peripheral blood of patients with PV.The number of DG3-specific IL-4+ Th2 cells per 5 × 105 PBMCs significantly decreased from 452.82 ± 199.29 before treatment to 241.68 ± 160.60 after treatment in seven patients (t =2.48,P ＜ 0.05).There was no significant correlation between specific Th cells,memory B cells and anti-Dsg3 antibody titers (all P ＞ 0.05).Conclusions The peptide DG3 (96-112) has pathogenic epitopes which could be recognized by specific Th cells of patients with PV.Both antigen-specific IFN-γ+ Th1 cells and IL-4+ Th2 cells play certain roles in the pathogenesis of pemphigns vulgaris,and IL-4+ Th2 cells appear to be more important.

AIM: To investigate the effect of problem - based learning ( PBL) used in the teaching of medical students'evidence-based medicine ( EBM) .
METHODS: Five classes ( total 147 students ) were randomly selected as experimental ( PBL ) group, at the same time, another 5 classes ( total 149 students ) were also randomly selected as control group, using traditional teaching method ( lecture-based learning, LBL ) in 2010 grade. The final examination scores of the experimental group were compared with control at the end of term. In addition, all students were interviewed using self -administered questionnaire to obtain their evaluation for PBL practice. SPSS13. 0 software was used for statistical analysis.
RESULTS: The homogeneity test in baseline survey showed that the basic characteristics between the two groups of students were no significant differences, and were comparable (P>0. 05). Final exam results showed that in addition to the scores of the EBM basic knowledge indicated no significant difference between two groups of students (P>0. 05), for the 5 steps of EBM procedure, namely, asking questions, finding the best evidence, evaluating the evidence, using and practicing the evidence, re - evaluating the evidence, and the total scores between the two groups, there were significant statistically differences (P<0. 05). The results to student learning evaluation showed that there was no significant difference between the two groups (P>0. 05) in aspects of better understanding classroom knowledge, improving language expression ability, and writing skill exercises. And other residual items had a significant difference ( P<0. 05), especially in aspects of improving enthusiasm for learning, self - study ability, improving learning efficiency, information analysis and utilization ability, team collaboration, and communication between teachers and students, however, there was a very significant difference (P<0. 001) between the two groups.
CONCLUSION: PBL teaching mode can effectively improve teaching effectiveness and the quality of EBM teaching, so the this teaching mode is worth further popularizing.

Objective To research the fibrosis characteristics of Atrial Fibrillation(AF) patients under microscope and its effect to post-operative sinus conversion of Radiofrequency ablation treatment.Methods Based on Lefi Atrial Dimension (LAD).Left atrial appendage tissue samples of 60 AF patients who received bipolar radiofrequency ablation treatment during open heart surgery between July and December 2011,were divided into 4 groups:group a (LAD≤50 mm),group b (50 mm ＜ LAD≤60 mm),group c (60 mm＜ LAD ≤70 mm),group d (LAD ＞ 70 mm),15 patients in each group.The levels of atrial muscle fibrosis and cell size differences are compared for colored slides of samples.Research are done for the difference of sinus rhythm restoration rate among the 4 groups based on ECG records at immediate,leaving hospital,and 3,6,12 months postoperative are collected for all groups.Results Differences of CVF and atrial muscle cell size measured under microscope are statistically meaningful(P ＜ 0.001).Sinus rhythm restoration rate differences at 6 month(P =0.039) and 12 month (P =0.037) post-operative are statistically meaningful.Further,immediate,leaving hospital,and 3,6,12 months postoperative sinus rhythm restoration rates are 93.3％,93.3％,93.3％,100％,100％ for group a,80.0％,73.3％,80.0％,80.0％,80.0％ for group b,66.7％,66.7％,66.7％,73.3％,66.7％ for goup c,and 53.3％,53.3％,53.3％,60.0％,60.0％ for group d respectively.Conclusion For rheumatic heart valve diseases patients who at 6 and 12 month post-operative,the greater the LAD,the higher the atrial fibrosis level,the greater the cell size,the lower the sinus rhythm restoration rate is.

OBJECTIVE: To analyze the features of drowning victims with psychiatric disorders to support further investigation on such cases. METHODS: One hundred and five archives of drowning victims with psychiatric disorders from January 2010 to June 2014 in Harbor Public Security Subbureau, Shanghai Public Security Bureau were reviewed. The decedents' general information, autopsy findings, case investigation data and disease histories were statistically analyzed. RESULTS: Of the 105 cases, 104 were suicides. Most decedents were poorly educated. Most suicides occurred in warm climate, and the corpses were usually found within 2 days. Most decedents had no suicide history and notes, but had abnormal mental or behavior changes before suicide. Depression, depression status and schizophrenia were the main types of psychiatric disorders, and only 9 decedents had received regular therapies before their deaths. CONCLUSION Mental disorder is a high risk factor in the drowning victims. Guardians should be aware of psychotic abnormal behaviors, especially during warm climate seasons.
China/epidemiology* ; Depression/epidemiology* ; Depressive Disorder/epidemiology* ; Drowning/psychology* ; Female ; Humans ; Male ; Mental Disorders/psychology* ; Psychotic Disorders ; Retrospective Studies ; Risk Factors ; Schizophrenia ; Seasons ; Suicide/statistics & numerical data*

OBJECTIVE: To compare the correlation between contrast vision (LV) and sweep visual evoked potential acuity (SVEP-A) among people with emmetropia, mild myopia, and moderate myopia. METHODS: The CV and SVEP-A were tested individually in 96 eyes from healthy young volunteers, including 37 eyes of emmetropia, 27 eyes of mild myopia, and 32 eyes of moderate myopia. The statistic analysis was done by ANOVA analysis and rank sum test. RESULTS: (1) With the decrease of contrast, CV and SVEP-A decreased in every group. (2) At 100% contrast, the difference of CV between emmetropia and mild myopia had statistical significance (P<0.05). At 100%, 25% and 10% contrast, the difference of CV between emmetropia and moderate myopia had statistical significance (P<0.05). (3) In the same group, the difference of 100% and 25% contrast had statistical significance (P < 0.05). So was between 100% and 10% contrast. (4) At 100% and 10% contrast, the difference of CV and SVEP-A had statistical significance (P < 0.05). CONCLUSION The CV of myopia relates to many factors including ametropia and fundus lesions. The correction of ametropia is important to the values of CV and SVEP-A.
Evoked Potentials, Visual/physiology* ; Eye ; Fundus Oculi ; Humans ; Myopia/physiopathology* ; Neurologic Examination ; Severity of Illness Index ; Vision Tests/instrumentation* ; Visual Acuity/physiology*

To investigate the biological properties of cryopreserved dendritic cell (DC) derived from K562 cell line, thus to provide a simple, quick and efficient preservative method of DC for infusion of DC to patients with leukemia after complete remission, fresh DC induced from K562 cell line (K562-DC) was frozen in -196 degrees C liquid nitrogen and -80 degrees C mechanical freezer by method of steps (RPMI 1640 with 10% DMSO, 20% FCS as cryopreservatives), and thawed in different time, respectively. Survivals of cryopreserved and fresh K562-DC, expression of surface antigens, stimulating index (SI) and cytotoxic eliminating rate were detected. The results of fresh induced cells were compared with that of cryopreserved ones. The results showed that before and after frozen in liquid nitrogen, the morphological characteristics of K562-DC had no distinct change; and both their expression rates of surface molecular and capacity to stimulate allogeneic lymphocyte had no statistic significance (P > 0.05). In addition, there were no differences in terms of viability, stimulatory capacity and cytotoxicity of K562-DC from two ways for less than one month (P > 0.05), but there were differences when frozen for more than one month (P < 0.05). It is concluded that there is no significant difference when frozen less than one month between liquid nitrogen and -80 degrees C freezer; but when time is more than one month, K562-DC frozen in -196 degrees C liquid nitrogen is better than that in -80 degrees C freezer.
Antigens, Surface ; immunology ; Cell Shape ; Cell Survival ; Cells, Cultured ; Cryopreservation ; methods ; Dendritic Cells ; immunology ; pathology ; Humans ; K562 Cells ; T-Lymphocytes, Cytotoxic ; immunology ; pathology ; Time Factors

The chemical constituents of 95% ethanol extract of Melastoma dodecandrum were isolated and purified by chromatography on silica gel, Sephadex LH-20, and HPLC, to obtain thirteen compounds eventually. On the basis of their physico-chemical properties and spectroscopic data, these compounds were identified as quercetin (1), quercetin-3-O-β-D-glucopyranoside (2), quercetin-3-O-(6"-O-p-coumaroyl) -β-D-glucopyranoside (3), kaempferol (4), kaempferol-3-O-β-D-glucopyranoside (5), kaempferol-3-O- [2",6"-di-O-(E)-coumaroyl]-β-D-glucopyra-noside (6), luteolin (7), luteolin-7-O-(6"-p-coumaroyl) -β-D-glucopyranoside (8), apigenin (9), apigenin-7-(6"-acetyl-glucopyranoside) (10) , naringenin (11), isovitexin (12), and epicatechin-[8,7-e] -4β-(4-hydroxyphenyl)-3,4-dyhydroxyl-2(3H)-pyranone (13). Eight compounds(3,5,6,8-11 and 13) were obtained from M. dodecandrum for the first time.
Apigenin ; analysis ; Chromatography ; methods ; Chromatography, High Pressure Liquid ; Dextrans ; Flavanones ; analysis ; Flavonoids ; analysis ; chemistry ; Glycosides ; analysis ; chemistry ; Kaempferols ; analysis ; Luteolin ; analysis ; Magnoliopsida ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analysis ; Silica Gel

Objective:To express the glycoprotein D of herpes simplex virus type 2 (gD2) in the insect cells,and to determine its immunogenicity.Methods:HSV-2 genome was used as the template for amplification of gD2 extracellular domain fragment gene by PCR.The PCR product was inserted into the vector Bacmind,and the constructed recombinant plasmid gD2-Bacmind was transfected into the sf9 cells to package the recombinant baculovirus.The Sf9 cells were infected by recombinant baculovirus seed derived from the forth passage(P4),the titer of P4 recombinant baculovirus was detected by a plaque assay and the expression of recombinant protein gD2 was determined by Western blotting method.The supernatant of infected cells was collected and purified by Ni-NTA affinity chromatography to obtain the target protein gD2,the purified gD2 protein was used to immunize the BALB/c mice in 0, 2, 4 weeks (gD2 group),and PBS was used as negative control(PBS group);the titers of gD2 specific IgG in serum were detected by ELISA assay.Results: The PCR analysis and sequencing results proved that gD2-Bacmind was constructed correctly.The titer of recombinant baculovirus was 2.0×109 pfu·mL-1,the purified gD2 was about 37 000 with expectation,the percentage of gD2 in total protein was 90%.The average value of Log10 of titer of gD2 specific IgG in serum detected by ELISA assay in gD2 group at the sixth week was 4.34,and there was significant difference compared with PBS group(P<0.01).Conclusion: The gD2 expressed by insect-baculovirus expression vector system has the immunogenicity and can be selected as candidate protein for HSV-2 vaccine.