ObjectiveTo assess the relationship of filaggrin expression with atopic diathesis and disease severity in patients with alopecia areata (AA).MethodsThirty-seven patients with AA aged (26.3 ± 10.6) years were enrolled in this study.Atopic diseases were noted in 8 of these patients.Clinical data and laboratory test resuhs were reviewed.Immunohistochemical staining was performed to quantify the expression of filaggrin protein in scalp biopsy specimens from all of the 37 patients with AA and from 10 human controls,and fluorescence-based semiquantitative reverse transcription-PCR to detect the expression of filaggrin mRNA in scalp biopsy specimens from 22 patients with AA and 13 healthy controls.Data were statistically analyzed by Mann Whitney U test,chi-square test,and Spearman's rank correlation test.ResultsThe expressions of filaggrin protein and mRNA were significantly lower in patients with AA than in the controls(P ＜ 0.05 or 0.01 ),and the decrease seemed more obvious in patients with large areas of lesions,long duration of disease,and nail abnormalities,but the degree of decrease was unrelated to the complication with atopic diseases.No significant differences were observed in sex ratio,age at onset,disease duration,area of hair loss,the prevalence of family history or incidence of nail abnormalities and increase in serum IgE and eosinophils,between patients with atopic diseases and those without.ConclusionsThe expressions of filaggrin protein and mRNA are decreased in patients with AA,suggesting that filaggrin may participate in the development of AA and is correlated with the severity of AA.

Objective To detect the expressions of apoptosis-related factors and inflammatory cytokines in superficial and deep layers of as well as anagen hair follicles in lesions of early alopecia areata (AA).Methods Scalp biopsy samples were collected from 25 patients with early AA and 15 healthy human controls.Fluorescence-based quantitative PCR was performed to detect mRNA expressions of apoptosis-related genes p53,caspase 3,Fas,survivin and bcl-2,as well as those of inflammatory cytokines interleukin (IL)-4,IL-10,IL-12 and interferon (IFN)-γ.An immunohistochemical assay was conducted to measure the expression of p53 protein in anagen hair follicles.Results Compared with control skin samples,anagen hair follicles in AA lesions showed significantly increased mRNA expression levels (expressed as 2-△△Ct) of pro-apoptotic factors caspase 3,p53 and Fas (6.78,8.01,9.74,respectively,all P ＜ 0.05),but decreased mRNA expression levels of antiapoptotic factors bcl-2 and survivin (0.08 and 0.03 respectively,both P ＜ 0.01),and similar mRNA expression levels of inflammatory cytokines.There was a significant increase in mRNA expression levels of Th1 cytokines IFN-γ and IL-12 (2.75 vs.1.00,P ＜ 0.05; 85.67 vs.1.00,P ＜ 0.01),but a significant decrease in the expression level of the Th2 cytokine IL-10 (0.002 vs.1.000,P ＜ 0.01) in superficial layers of AA lesions compared with those of normal control skin.The degree of changes in mRNA expression levels of IL-10 and IL-12 was significantly higher in superficial layers than in deep layers of AA lesions (P＜0.01 and 0.05 respectively).The immunohistochemical assay showed that the number of p53-positive cells per 100 cells in anagen hair follicles of AA lesions was higher than that in those of control skin (t =23.79,P ＜ 0.01).Conclusions Anagen hair follicles in AA lesions exhibit high expressions of pro-apoptosis factors,but low expressions of antiapoptotic factors,suggesting that apoptotic factors play a role in the occurrence of AA.

Purpose:Taking tuberculosis as an example, this paper aims at to analyzing the level of reimburse-ment for infectious diseases care, and clarifying the government responsibility. Methods:In order to achieve the ob-jective of this research, UHC framework was used to analyze the security level. Result:The findings of this research reveal that TB in-patients' Compensation Ratio of the New Cooperative Medical Scheme ( NCMS) was lower than aver-age level of all the NCMS patients, the out-patients' was even lower. The categories of anti-tuberculotic for free was limited, the utilization was not as expected. Medical assistance covered few people in spite of its high level of reim-bursement. Conclusion:Based on the findings of this review, it has been revealed that the medical insurance didn't make a big difference in financial protection for patients with infectious diseases. As the treatment for of infectious diseases is a quasi-public good, the government has to shoulder the responsibility of improving the compensation ratio of the patients.

Objective To observe the microstructural changes in lesions of alopecia areata (AA) with dermoscopy and to evaluate their correlation with clinicopathological manifestations. Methods The area of alopecia of 62 patients with AA and 44 patients with other types of hair loss were observed by using a noncontact polarized dermoscope (Dermlite, USA). Clinical data on and laboratory findings from these patients were collected. Pathological examination was carried out with scalp biopsy specimens from the alopecia area of 15 AA patients. Results Characteristic dermoscopic signs of AA included yellow dots, black dots, broken hairs, exclamation mark hairs, short vellus hair and newly-grown short hairs. Among these signs, yellow dots showed the highest prevalence (83.9%). Exclamation mark hairs, black dots and broken hairs were rather specific signs for AA, and the prevalence of the three signs was positively correlated with disease activity and positivity rate of hair-pull test. A positive correlation was also noted between the prevalence of elevated thyroid peroxidase antibody levels and positivity rate of hair-pull test (r = 0.269, P ＜ 0.05 ) as well as prevalence of broken hairs (r = 0.445, P ＜ 0.05), and between the prevalence of yellow dots and that of keratinous plug in follicular orifice. There was a negative correlation between the prevalence of newly-grown short hairs and perifollicular mast cell infiltration and between the prevalence of black dots and the anagen/catagen ratio. Conclusions Yellow dots can serve as a preliminary screening marker for AA. Exclamation mark hairs, black dots and broken hairs are highly sensitive for the confirmation of diagnosis of AA, and often predict progressive AA.Dermoscopic signs are well correlated to the histopathology features of AA, and may be useful for the evaluation of disease severity and guidance on the treatment of AA.

In order to study the anti-viral mechanism of extracted ZG from Gardenia, the effect of extracted ZG on Hep-2 cell membrane potential, Na -K+-ATPase activity and membrane fluidity post infected with parainfluenza virus type 1 (PIV-1) was observed. Acetylcholine which was fluorescent labeled with DiBAC4 (3) was taken as positive control to observe the changes of membrane potential and was measured by flow cytometer. The phosphorus determination method and spectrophotometer were used to measure the Na+-K+-ATPase activity of Hep-2 cell membrane post PIV-1 infection. Hep-2 cell membrane phospholipids was labeled with fluorescent NBD-C6-HPC and membrane fluidity was measured by confocal laser scanning microscope. The results demonstated that after PIV-1 infection the Hep-2 cell membrane potential decreased significantly and the membrane was in the state of hyperpolarization, Na+-K+-ATPase activity increased and membrane fluidity decreased significantly. There was no apparent interferring effect of extracted ZG on the changes of membrane potential and Na+-K+-ATPase activity post PIV-1 infection, while membrane fluidity was improved significantly. Acetylcholine improved the state of hyperpolarization. The changes of membrane potential, Na -K+-ATPase activity and membrane fluidity might be the biomechanism of PIV-1 infectoin. The extracted ZG improved membrane fluidity to prevent from PIV-1 infection by protecting the cell membrane, which was probably the mechanism of anti-PIV-1 activity of the extracted ZG, but ZG probably had nothing to do with membrane potential and Na+-K+-ATPase activity.
Acetylcholine ; pharmacology ; Antiviral Agents ; pharmacology ; Cell Line, Tumor ; Cell Membrane ; drug effects ; Gardenia ; chemistry ; Humans ; Membrane Fluidity ; drug effects ; Membrane Potentials ; drug effects ; Parainfluenza Virus 1, Human ; drug effects ; Plant Extracts ; pharmacology ; Sodium-Potassium-Exchanging ATPase ; metabolism

The study was aimed to isolate and establish mesenchymal stem cell line from adult murine bone marrow as well as to identify its biological characteristics and differentiation potential. Bone marrow cells (BMCs) were collected by flushing the femurs and tibias of 4 - 5-week-old male C57BL/6 mice, and were inoculated at a concentration of 1 x 10(6)/cm(2). mMSCs were isolated, enriched and expanded by using bone marrow adherant culture and monoclonal culture. The characteristics of the cells, such as morphology, growth pattern, cell cycle, phenotype, karyotype and multipotent differentiation potential, cytogenetic stability and tumorigenesis were determined. The results indicated that the cell population consisted of spindle- and star-shaped cells, they were highly positive for CD29, CD44, Sca-1, MHC-I, moderate positive for CD13, CD90.2 and negative for CD117, CD45, Flk-1 and MHC-II. mMSCs could be induced to differentiate into adipocytes, osteoblast cells and chondrocytes. It is concluded that mMSC can be isolted, expanded and enriched by using bone marrow adhcrent culture and monoclonal culture. No tumor formations are observed for 3 months in nude mice after subcutaneous injection. mMSCs retain their properties after at least 30 passages in culture as well as from frozen stocks.
Animals ; Cell Differentiation ; physiology ; Cell Proliferation ; Cell Separation ; methods ; Cells, Cultured ; Hyaluronan Receptors ; metabolism ; Integrin beta1 ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; immunology ; physiology ; Mice ; Mice, Inbred C57BL ; Mice, Nude

OBJECTIVETo explore the clinical and laboratory characteristics of myleodysplastic syndrome (MDS)/myeloproliferative neoplasm (MPN) with PDGFRβ abnormalities.

METHODSChromosome specimens were prepared directly and/or short-time culture of bone marrow cells. Karyotyping was performed with R-binding technique. Fluorescence in situ hybridization (FISH) was performed using PDGFRβ, PDGFRα, FGFR1 break-apart probes and whole chromosome 5 and 12 painting probes, respectively. The expression of JAK2 V617F was measured with quantitative PCR.

RESULTSThe clinical and hematological findings of 27 patients were compatible with diagnosis of MDS/MPN. PDGFRβ rearrangement was detected in 4 patients with D-FISH, and 2 of which were confirmed as t(5;12) by chromosome painting. PDGFRα, FGFR1 and JAK2 V617F mutation were not detected in these 4 PDGFRβ positive MDS/MPN patients with.

CONCLUSIONSPDGFRβ gene rearrangement may be detected in some MDS/MPN patients. FISH is a convenient and reliable approach to detect PDGFRβ gene.

Two clinical ablation protocols,2C3L and stepwise,have been routinely used in our group to treat atrial fibrillation (AF),but with a less than 60％ long-term arrhythmia-free outcome achieved in patients.The goal of this study was to examine the underlying mechanism of low success in clinical outcome.MRI images from one patient were used to reconstruct a human atrial anatomical model,and fibrotic tissue was manually added to represent the arrhythmia substrate.AF was induced with standard protocols used in clinical practice.2C3L and stepwise were then used to test the efficacy of arrhythmia termination in our model.The results showed that re-entries induced in our model could not be terminated by using either 2C3L or the stepwise protocol.Although some of the induced re-entries were terminated,others emerged in new areas.Ablation using only the 2C3L or stepwise method was not sufficient to terminate all re-entries in our model,which may partially explain the poor long-term arrhythmiafree outcomes in clinical practice.Our findings also suggest that computational heart modelling is an important tool to assist in the establishment of optimal ablation strategies.

Objective To improve the accuracy of patient' s identification and normalize the checking details for preventing from the error intravenous ( i.v.) infusion.Methods Participants who were eligible for this study from Department of Orthopedics,Nanxishan Hospital of Guangxi Zhuang Autonomous Region were assigned to control group and observational group by random number method.The control group included 1 410 patients which had 42 026 set intravenous infusion solutions and confirmed identity by the traditional procedure.The observation group included 41 102 set intravenous infusion solutions in 1 303 patients.In observational group,we confirmed patients' identity by speaking out their name and age and looking over the patient' s label on the infusion bottle by their family members along with the traditional procedure.The number of patients and i.v.solutions set regarding borderline errors of identification during venepuncture or replacing infusion bottle were investigated in two groups.Results 1 404 patients in control group were accurately confirmed identity and 6 patients had borderline errors.1 303 patients in observation group were accurately confirmed identity and 0 patients had borderline errors,the difference between the two groups showed statistical significance ( P =0.032).In control group,42 020 set were accurately confirmed identity and 6 set had borderline errors,however,in observation group 41 102 set were accurately confirmed identity and 0 set had borderline errors,the difference between the two groups showed statistical significance ( P =0.031 ).Conclusions Patients participate in double-identity confirmation can effectively prevent from error of i.v.solutions.

Objective To investigate the clinical relevant factors causing laryngeal stenosis after partial laryngectomy and the its nursing intervention. Methods A retrospective study was carded out to review the history clinical data from 138 patients of partial laryngectomy in the First Affiliated Hospital of Sun Yat-Sen University between January 1994 to October 2004. The clinical relevant factors causing laryngeal stenosis included age,sex,TNM stage, original position tumor, resects extension of thyroid cartilage, resects extension of larynx parenchyma, repair model, larynx pattern, duration of using bacteriophage postoperative, postoperative radiotherapy, lung infection postoperative, gastroesophageal reflux, diabetes. Multivariate stepwise logistic regression model was used for the analysis. Results Among 138 cases after partial laryngectomy, stenosis developed in 25 cases, incidence rate was 18.12%. In multivariate analysis,it was confirmed that the following variable correlated to laryngeal stenosis, resects extension of thyroid cartilage, postoperative radiotherapy, lung infection postoperative, and gastroesophageal reflux. Conclusions The clinical relevant factors causing laryngeal stenosis after partial laryngectomy were various. Statistical analysis showed that resects extension of thyroid cartilage, postoperative radiotherapy, lung infection postoperative, gastroesophageal reflux were risk factors which may cause laryngeal stenosis. The prophylactic nursing interventions should be taken to cut down the rate of laryngeal stenosis after partial laryngectomy and improve the quality of life.