Objective To study the effect of human ?-globin matrix attachment region(MAR) on transgene expression in stably transfected CHO cells.Methods Expression vector was constructed,which contained the ?-globin MAR in both sides of Chloramphenicol acetyltransferase(CAT) reporter gene expression cassette in cis,then transfected into CHO cells.The CAT reporter gene expression was analyzed by ELISA method.Results The ?-globin MAR enhanced the CAT gene expression 5.5-fold in stably transformed CHO cells,while the transgene expression variation among individuals of transformants was decreased.Conclusion MAR increase transgene expression in stably transfected CHO cells.

Objective To profile the intraspecific type of Trichophyton mentagrophytes clinically isolated from different anatomical sites of patients, and to compare the performance of different target sites for the identification of Trichophyton mentagrophytes complex strains. Methods A total of 48 Trichophyton mentagrophytes strains, which were clinically isolated from Department of Dermatology, Wuhan No. 1 Hospital in the latest 3 years, were included in this study. The phenotypes of these Trichophyton mentagrophytes isolates were primarily determined by morphological observation and the urease test. PCR was performed to amplify the nuclear ribosomal internal transcribed spacer(ITS) region and the D1?D2 domains of the large?subunit ribosomal DNA(28S rDNA)followed by DNA sequencing. Then, Clustal X2 software and MEGA 6.0 software were used to analyze the ITS and D1?D2 sequences and to build phylogenetic trees by the maximum?likelihood method (bootstrap = 2000). Results As the ITS sequence?based phylogenetic tree showed, the probability that the 48 isolates were grouped into the Trichophyton interdigitale clade reached 92%. However, Trichophyton interdigitale could not be effectively differentiated from Trichophyton quinckeanum by the D1?D2 sequence?based phylogenetic tree. In addition, Trichophyton interdigitale showed various appearances, including woolen type, downy type, cream type, powdery type and granular type. Conclusions Trichophyton mentagrophytes can be identified to the species level based on the sequence of ITS region, which shows higher efficiency in identifying Trichophyton mentagrophytes complex than the D1?D2 domains. Morphological characteristics can not serve as the basis for intraspecific typing of Trichophyton mentagrophytes.

AIM:To screen the gene expression profiles of normal gastric mucosa,primary gastric cancer and metastatic lymph nodes by gene expression microarray and the associated genes with lymph node metastasis by bioinformatics. METHODS:The differentially expressed genes of nontumorous gastric mucosa (group A),primary gastric cancer (group B) and metastatic lymph nodes (group C) were screened by gene expression microarray obtained from Affymetrix company. The results were further analyzed by bioinformatic software including Gene Expression Profiles Analysis of Cohort Experiment,Gene Ontology Enrichment Analysis and Pathway Significant Analysis. The expression of TLN1 in group A,B and C were confirmed by real-time reverse transcription PCR. RESULTS:278 genes were persistent up-regulated in group A,B,C,which participated mainly in immunologic responses,cell adhesion,phosphate transport,inorganic anion transport,cell chemotaxis,cell motility and signal transduction. While 387 genes were persistent down-regulated in group A,B,C,which were concerned with digestion,glucide metabolism,lipid metabolism,protein metabolism,one-carbon compound metabolism,nitrogen compound catabolism and cell adhesion. The pathway analysis suggested that integrin-mediated cell adhesion pathway were abnormally regulated. These genes including THBS1,TLN,CAPN3,ITGAX,SORBS1,CAPN6,CAPN9 were continuous up-regulated or down-regulated in integrin-mediated cell adhesion pathway. The expressions of TLN1 in group A,B,C were 0.0000342?0.0000711,0.1064?0.1251 and 0.2886?0.3529,respectively. The expression of TLN1 in metastatic lymph nodes was significantly higher than that in nontumorous gastric mucosa and primary gastric cancer (P

Objective To explore short-term effect of tonsillotomy on children′s obstructive sleep apnea hypopnea syndrome (OSAHS) with tonsil hypertrophy by Ellman double-frequency radio-frequency machine .Methods Totally 30 cases children′s OS-AHS with tonsil hypertrophy were had tonsillotomy by Ellman double-frequency radio-frequency machine .They were compared with 30 cases children′s OSAHS with tonsil hypertrophy by traditional tonsillectomy .Short-term effect of tonsillotomy were ana-lyzed by wound bleeding ,operation time ,postoperative pain ,difficulty eating ,PSG testing ,immune function ,growth and develop-ment .Results Postoperative symptom in 1 weeks and 6 months ,compared with the traditional tonsillectomy ,30 cases of OSAHS patients were found that there were less bleeding ,shorter operative time ,mild postoperative pain ,no difficulty in feeding ,improving sleep-disordered breathing ,no decline in immune function ,good growth and development .Conclusion Short-term effect of tonsillot-omy on children′s OSAHS with tonsil hypertrophy is good by Ellman double-frequency radio-frequency machine ,it is a minimally invasive ,safe and effective surgical method children OSA HS .

Objective:This study evaluated the economical effects of two different regimens for metastatic breast cancer, namely, paclitaxel plus either carboplatin (TP) or epirubicin (TE). Methods:The cost-effectiveness method in pharmacoeconomics was adopted to analyze retrospectively the two different regimens. Results:The median follow up was 23.5 (range:9 to 42) months. The overall response rate for TP and TE were 78.33%and 80.00%, respectively. The 1-and 2-year progression-free survival rates of TP and TE were 43.6%and 38.9%and 10.8%and 17.4%, respectively. The 1-and 2-year overall survival rates were 80.3%and 78.3%, respectively for TP, whereas the corresponding values for TE were 53.2%and 47.9%. No statistically significant difference was found between the two groups (P>0.05). Cost-effectiveness analysis showed that the average costs of the TP and TE regimens were 10 303.8 and 13 853.3 yuan, respectively, with corresponding cost-effectiveness ratios of 131.54 and 173.17 (P<0.01). For the chemotherapy toxicity, the alopecia reactions of the TP group were significantly lower than those of the TE group (P<0.01). Conclusion:The short-and long-term efficacies of the two regimens were similar. TP regimen was the optimal scheme for advanced metastatic breast cancer.

Objective: To evaluate the pharmacokinetics of palonosetron hydrochloride in healthy volunteers. Methods: Thir-ty-one healthy volunteers were grouped into three palonosetron hydrochloride dosage regimens of 0.125, 0.25, and 0.5 mg. The plasma concentrations of palonosetron were determined by ultra high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). DAS 2.1 software was applied to assess the plasma concentration-time data. Results:After intravenous injection of 0.125, 0.25, and 0.5 mg palonosetron to the subjects, the AUC0-168h values of palonosetron were (7.5±2.5), (15.2±4.0), and (34.8±9.7) μg· h·mL-1. The t1/2 values were (27.2±9.5), ( 27.2±6.5), and (31.4±5.6) h. Palonosetron exposure increased proportionally with the dose range of 0.125 mg to 0.5 mg. The correlation coefficient was 0.998. No grade 3 or grade 4 toxicity was observed during the study. Con-clusion:A rapid, sensitive, and selective UPLC-MS/MS method for palonosetron quantification in human plasma was developed and validated. All the participants indicated high tolerance throughout the study. Our data showed that palonosetron exhibits linear pharma-cokinetics over the the dose range of 0.125 mg to 0.5 mg.

Objective To compare the reactive oxygen species (ROS) expression in human neutrophils phagocytizing Sporothrix Schenckii and Candida albicans yeast cells,and to compare the fungicidal activity of human neutrophils against Sporothrix Schenckii and Candida albicans.Methods Human neutrophils were isolated from peripheral blood by using density gradient centrifugation method,and cultured with the presence of the yeast phase of a Sporothrix Schenckii clinical isolate and a standard strain of Candida albicans (ATCC 90028)at a multiplicity of infection of 10 or 1 for 60-210 minutes.Subsequently,flow cytometry with ROS probe (2',7'-dichlorofluorescein diacetate,DCFH-DA) was carried out to for the real time detection of intracellular ROS level,confocal laser scanning microscopy (CLSM) for the observation of ROS distribution.In addition,the fungicidal efficiency of neutrophils against Sporothrix Schenckii and Candida albicans was estimated by the number of colonies after additional culture of neutrophil lysates on brain-heart infusion agar (BHIA) medium.Statistical analysis was done by using univariate analysis of variance and LSD-t test.Results The intracellular ROS level peaked at 60 minutes in neutrophils incubated with Sporothrix Schenckii yeast cells,then decreased rapidly from 60 minutes to 210 minutes.Compared with the neutrophils incubated with Candida albicans yeast cells,those with Sporothrix Schenckii yeast cells showed a higher ROS level (expressed as mean fluorescence intensity) at 60minutes (159.67 ± 11.34 vs.112.22 ± 9.66,P＜ 0.01),but a lower ROS level at 120 minutes (89.01 ± 9.81 vs.110.25 ± 7.28,P＜ 0.05) and 180 minutes (57.63 ± 8.46 vs.109.98 ± 9.00,P＜ 0.01).CLSM revealed that ROS was mainly distributed in neutrophils with phagocytized fungal spores,and especially on the surface of phagocytized spores.Furthermore,the percentage of yeast cells killed by neutrophils was significantly lower for Sporothrix Schenckii than for Candida albicans at 180 minutes (19.21％ ± 3.68％ vs.26.63％ ± 4.97％,P ＜ 0.01).Conclusions Differential expression of intracellular ROS was observed in neutrophils after phagocytosis of Candida albicans and Sporothrix schenckii.Neutrophils exert a stronger fungicidal activity against Sporothrix Schenckii in comparison with Candida albicans,which may be associated with the rapid decrease of ROS level in neutrophils after phagocytosis.

A 54-year-old female farmer presented with a pea-sized red nodule on the left upper limb near the wrist for 15 days,which occurred after trauma,gradually became swollen and ruptured,and developed into multiple nodules arranged in a chain in 30 days.Skin examination revealed multiple hard purple-red nodules arranged in a line on her left upper limb,some of which were ruptured with a small amount of purulent exndate.Histopathological examination further revealed that the focus of infection manifested as pyogenic granuloma-like inflammation mainly infiltrated with mixed inflammatory cells.Periodic acid Schiff (PAS) staining showed no fungal structures,including fungal spores,hyphae and asteroid body.The biopsy tissue culture yielded the fungus.According to the morphological analysis of the cultures and results of molecular identification based on internal transcribed spacer (ITS) and calmodulin (CAL) coding regions,this case was finally diagnosed as lymphocutaneous sporotrichosis caused by Sporothrix schenckii sensu stricto.The patient was treated with oral potassium iodide 10％ solution in a dose of 10 ml thrice a day.After 2-month treatment,the patient felt that the lesions were obviously improved,but afterwards she was lost to follow-up.This research report suggests that phenotypic analysis combined with ITS/CAL-based molecular identification can accurately identify Sporothrix schenckii complex at the species level.

Objective To study Twist,E-CD expression in colorectal cancer tissues and its relationship with colorectal cancer invasion,metastasis and prognosis.Methods Immunohistochemical staining (EnVision) was used to detect E-CD,Twist expression of normal colon mucosa in 30 cases,colorectal adenoma in 30 cases and colorectal cancer tissues in 142 cases.Chi-square、Fisher's and Spearman test were used to analyze E-CD and Twist protein expression,Kaplan-Meier survival analysis and multivariate COX regression were used to analyze prognosis of patients.Results E-CD in the normal mucosa were positively expressed in 90％ cases,which was significantly higher than that in colorectal adenomas (63％) (P =0.046) and colorectal cancer tissues (42％) (P =0.000).E-CD expression was related to tumor differentiation (P =0.048),invasion depth (P =0.000),vein (P =0.000) and lymph vessel invasions (P =0.030),lymph node metastasis (P =0.001) and Dukes' stage (P =0.016),but not related to patient's age(P =0.174),gender(P =0.159),tumor size (P =0.628) and tumor histological type (P =0.153).1,3,5 year survival rate in patients with positive E-CD expression was significantly higher than that in patients with negative expression (P =0.000).Positive expression rate of Twist in colorectal cancer tissues (68％) was significantly higher than that in normal mucosa (20％,P =0.000) and colorectal adenomas (30％,P =0.000).Twist expression was related to tumor histological type (P =0.000),differentiation(P =0.000),invasion depth(P =0.000),vein(P =0.000) and lymph vessel invasions(P =0.000),lymph node metastasis(P =0.010) and Dukes' stage(P =0.000).1,3,5 year's survival rate of Twist-negative expression patients was significantly higher than that in patients with positive expression (P =0.000).E-CD and Twist in colorectal cancer tissues were negatively correlated (r =-0.530,P =0.000).COX multivariate analysis shows that vein invasion (P =0.045),lymph node metastasis (P =0.040),Dukes' stage (P =0.000),E-CD (P =0.003) and Twist (P =0.031) were independent prognostic indicators.Conclusions E-CD and Twist expression in colorectal cancer are related to tumor invasion,metastasis and prognosis.Low E-CD expression and high Twist expression are related to poor prognosis of colorectal cancer patients.

Objective To investigate the mechanism of soluble β-1, 3-D-glucan in G-test positive serum in inhibiting ROS-dependent killing of Candida albicans ( C.albicans ) mediated by neutrophil Dectin-1.Methods The expression and distribution of internalized Dectin-1 and triggered ROS in human neutrophils were detected by using confocal/two-photon laser scanning microscopy upon stimulation with C.albicans (MOI=10) which was pretreated with β-1, 3-D-glucanase (10 U/ml) or not.Abrogation test was used to analyze whether intracellular Dectin-1 was involved in C.albicans-triggered ROS production in human neutrophils.Furthermore, flow cytometry analysis was performed to detect the expression of intracel-lular Dectin-1 and ROS in neutrophils which were pretreated respectively with G-test positive serum at differ-ent dilutions for 60 min and then stimulated with C.albicans for another 60 min at 37℃.Results After stimulated with C.albicans (MOI=10) for 60 min, the expression of Dectin-1 in neutrophils was recruited to the spores of opsonophagocytized C.albicans, and partly co-localized with the triggered ROS production . However, the expression of intracellular Dectin-1 was not observed in neutrophils when stimulated with β-1, 3-D-glucanase pretreated C.albicans for 60 min at 37℃.Abrogation test further showed that C.albicans-trig-gered ROS production in neutrophils was partly and irreversibly inhibited by adding Dectin -1 blocking mAb of 5 μg/ml.In addition , both the triggered expression of intracellular Dectin-1 and ROS production in neu-trophils stimulated with C.albicans ( MOI=10 ) in the presence of G-test positive serum were significantly lower than those of neutrophils stimulated only with C.albicans (LSD-t test, P<0.01).Linear regression a-nalysis suggested that the triggered intracellular Dectin-1 and ROS production in neutrophils upon stimulation with C.albicans were both inhibited by soluble β-1, 3-D-glucan in a dose-dependent manner (Dectin-1,R2=0.702,P<0.01;ROS,R2=0.588,P<0.01 ).Conclusion Taken together, these results demonstrated that the soluble β-1, 3-D-glucan in G-test positive serum may play a role in inhibiting the ROS-dependent killing of C.albicans by interfering with internalized expression of neutrophil Dectin-1.