Objective To prepare monoclonal antibody(McAb) against human tissue kallikrein (HK) and develop an ELISA kit allows for the in vitro quantitative determination of human tissue kallikrein in urine. Methods To generate a monoclonal antibody specific for TK, the synthetic TK peptide consisting of 12 amine acids(12P), was fused to keyhole limpet hemocyanin(KLH) and used for immunization. Using hybridoma screening, monoclonal secreting cell lines were identified and used to generate ascites in BALB/c mouse. Antibody was purified by affinity column chromatography. 12% SDS-PAGE and Western blot were used to visualize the purified antibody. This kit employs indirect competitive ELISA technique and BiotinAvidin System. 12P was fused to bovine serum albumin(BSA) and has been pre-coated onto a microplate at first. Standards and samples were added to the appropriate microplate wells with a biotin-conjugated McAb croplate well. A TMB substrate solution is added to each well. The enzyme-substrate reaction is terminating by the addition of a sulphuric acid solution and the color change is measured spectrotometrically at a wavelength of 450 nm. The concentration of tissue kallikrein in the samples is then determined by comparing the O.D. of the samples to the standard curve. Results 8 hybridoma cell lines secreting mAbs special to HK,SDS-PAGE and Western blot demonstrated successful preparing and purification of McAb( 100% ). The linearity of this ELISA kit is demonstrated(r =0. 990). The range of detection of the assay is 0.008 μg/ml to 0. 5 μg/ml. The assay remained stable, with no change in the values measured, over five cycles of freezing and thawing. Conclusion 8 McAbs against HK have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for detecting HK can meet the needs of detection of HK in urine samples.

Objective To establish high resolution, reproducible 2-dimensional electrophoresis (2-DE) profiles of invasive and non-invasive pituitary adenoma tissues and to identify differentially ex-pressed proteins between the invasive and non-invasive tissues. Methods The proteome from invasive and non-invasive pituitary adenomas tissues was dissected and analyzed by: (1) immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis, (2) silver staining, (3) imageMaster 2-D software analysis, (4) peptide mass fingerprint based (PMS) on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and (5) database comparison. Results High-resolution 2-D patterns of invasive and non-invasive pituitary adenoma tissues were successfully produced and re-peated 3 times for each sample. An average of 1080±24 and 1035±28 spots were detected for invasive and non-invasive pituitary adenoma tissues, respectively. Additionally, 975±45 and 918±56 spots were found to have an average matching rate of 90.3% and 88.7% for invasive and non-invasive tissues, re-spectively. The spot positional deviation was (1.563±0.259) mm for IEF and (1.088±0.206) mm for SDS-PAGE. A total of 99 spots of differential expression were matched between the invasive and non-in-vasive pituitary adenoma tissues. Thirty differential proteins, some of which were involved in the regula-tion of cells cycle and signal transduction, were initially characterized by PMS. Conclusion The acquisi-tion of well-resolved and reproducible 2-D patterns of invasive and non-invasive pituitary adenoma tissues and the identification of differentially expressed proteins provides a proteome database for invasive pituita-ry adenomas.

Functional electrical stimulation(FES) offers a vast potential for partial restoration of parlayzed movements. In practice,understanding on the underlying mechanism and limitations of electrical activation of nerve is essential to guide a successful deployment of FES technology to clinical utilization. Thisthree part article will review the basic concepts,system design and applications of FES. In part I,conceptssuch as activation threshold,recruitment order are discussed. Part Ⅱ introduces stimulation waveforme,safeparameters and tissue damage,as well as the design principle of stimulators for use with percutaneous electrodes. Part Ⅲ outlines clinical applications of FES,in particular,for restoration of hand grasp function forC5/C6 patients.

Functional electrical stimulation(FES) offers a vast potential for partial restoration of parlayzed movements. This three part article will review the basic concepts,system design and applications of FES. In part I,concepts such as activation threshold, recruitment order are discussed. Part Ⅱ introduces stimulation waveforme,safe parameters and tissue damage,as well as the design principle of stimulators for use with percutaneous electrodes. Part Ⅲ outlines clinical applications of FES,in particular,for restoration of hand grasp function for C5/C6 patients.

Objective: To investigate the impact and its possible mechanism of hydrogen sulfide (H2S) on myocardial collagen remodeling in experimental rats with diabetic mellitus (DM).
Methods: Rat’s DM model was established by intraperitoneal injection of STZ at 40 mg/kg. A total of 40 SD rats were randomly divided into 4 groups:Control group, DM group, DM+NaHS group, in which NaHS worked as exogenous donor of H2S and NaHS control group. n=10 in each group, all animals were treated for 8 weeks. The cardiac collagen deposition was observed by Masson staining, protein expressions of cardiac collagen types I, III, IV and transforming growth factorβ1 (TGF-β1), connective tissue growth factor (CTGF) were examined by Western blot analysis.
Results: Compared with Control group, DM group showed increased protein expressions of cardiac collagen types I and III, up-regulated expressions of TGF-β1 and CTGF, P<0.05;while the expressions of collagen type IV were similar between 2 groups. Compared with DM group, DM+NaHS group presented reduced cardiac collagen expression, decreased expression of collagen types I and III, down-regulated expressions of TGF-β1 and CTGF, P<0.05;while the expressions of collagen type IV were similar between 2 groups.
Conclusion: H2S may improve the myocardial collagen remodeling in experimental DM rats, the mechanism might be related to the down-regulation of TGF-β1 and CTGF expression.

[ ABSTRACT] AIM:To explore the effects of hydrogen sulfide ( H2 S) on the myocardial fibrosis in a rat model of diabetes and its mechanism.METHODS:Single intraperitoneal injection of streptozotocin ( STZ) was utilized to establish a rat model of diabetes.Sodium hydrosulfide was used as an exogenous donor of hydrogen sulfide.Male SD rats were ran-domly divided into control group, STZ group, STZ+H2 S group and H2 S group.Eight weeks later, HE and VG staining methods were used to observe the collagen distribution and collagen volume fraction was measured by image analysis.The expression levels of type I collagen, PPARγand NF-κB in the cardiac tissues were determined by Western blotting.RE-SULTS:Compared with control group, collagen distribution and the expression levels of type I collagen and NF-κB in the cardiac tissues were markedly increased (P<0.05), while PPARγwas significantly decreased in STZ group (P<0.05), but these indexes were reversed significantly in STZ+H2S group (P<0.05).The expression levels of type I collagen, PPARγand NF-κB had no significant difference between H2 S group and control group.CONCLUSION:Hydrogen sulfide attenuates cardiac fibrosis in diabetic rats, and its mechanism may be related to PPARγ-NF-κB signaling pathway.

ObjectiveTo observe the effect of light therapy combined with fluoxetine on winter attack depression.Methods68 patients of winter attack depression were divided randomly into the treatment group and control group with 34 cases in each group. Patients of the treatment group were treated by bright white light combined with fluoxetine (20 mg/d), but cases of the control group were treated only with fluoxetine. Therapeutic effects of two groups were evaluated with BDI, HAMD, and SCL-90 scales and traditional evaluation.ResultsPatients of the treatment group had a better effect than that of the control group (P<0.05).ConclusionLight therapy combined with fluoxetine has a good short and long term effect on winter attack depression, and can prevent from recurrence.

Objective To establish a stable and efficient method of culturing imDCs in vitro,and to explore the effect of GW5074, which blocks ERK1/2 signal pathway in the process of imnature dentritic cells (imDCs) on inducing differentiation of the na(i)ve allogeneic CD4+ T cells into Treg cells in vitro. Methods The imDCs and mature DCs (mDCs) were isolated and cultured from the peripheral blood mononuclear cells (PBMC) derived from a healthy adult male volunteer, and they were identified by cell morphology, cell surface marker and cell functions respectively. Na(i)ve CD4+ T cells were isolated from newborn umbilical vein blood and were divided into 5 groups to be cultured: (1) Blank control group: Na(i)ve CD4+ T cells were cultured alone;(2) Positive control group: The irrDCs were Middle-concentration GW5074 group;(5) High-concentration GW5074 group. In the last three groups, imDCs and na(i)ve CD4+ T cells were co-cultured, the same as the positive control group, but these groups were added by GW5074 dilution at the concentrations of 8, 24, and 40μmol/Lrespectively. After co-culture for 5 days, the transformation ratio from naive CD4+T cells to Treg T cells was detected by flow cytometry. Results On the surface of imDCs, there was stronger pression of CD1a, but weaker expression of CD80 and CD83. On the contrary, on the surface of mDCs, there was weaker expression of CD1a, but stronger expression of CD80 and CD83. The stimulation index in imDCs group and mDCs group was 1.12±0.03 and 2.85±0. 07 respectively. The transformation ratio of Treg T cells in blank control group, positive control group, low-concentration GW5074 group, middle-concentration GW5074 group and high-concentration GW5074 group was (5. 81±1.36)%, (35.73±2.07)%, (22.53±2.11)%, (11.55±1.73)%, and (4.97±1.83)%respectively. One-way ANOVA analysis revealed that there was no significant difference between high-concentration GW5074 group and blank control group, P＞0. 05, but significant difference between the remaining groups, P＜0.01. Conclusion High purity of imDCs can be obtained from PBMC by induction with rhGM-CSF and rhIL-4. ERK1/2 signal pathway plays a role in inducing the immune tolerance. GW5074 can inhibit differentiation of na(i)ve CD4+ T cells into Treg T cells.

OBJECTIVE: To explore whether oncogenes DJ-1 and HSP27 are associated with invasiveness of human pituitary adenoma. METHODS: Total proteins were extracted from samples of 20 invasive and 20 non-invasive pituitary adenomas and the expression of DJ-1 and HSP27 was analyzed by Western blot. The correlation of DJ-1and HSP27 with the invasiveness of pituitary adenoma was analyzed. RESULTS: The strong positive rates of DJ-1 and HSP27 in the 20 invasive pituitary adenoma were 70% (14/20) and 80% (16/20), respectively. The invasive group had significantly higher expression of DJ-1 and HSP27 proteins than the noninvasive group [10% (2/20), 10% (2/20), respectively]. There was a positive correlation between the expression of DJ-1, HSP27 proteins and the invasiveness of pituitary adenoma as judged by the Spearman rank correlation test (P<0.05). CONCLUSION The proliferative activity and abnormal expression of oncogenes DJ-1 and HSP27 may play a significant role in tumorigenesis and progression of pituitary adenoma. There was a significant correlation between the expression of DJ-1 and HSP27 and the invasiveness of pituitary adenoma.
Adenoma ; metabolism ; pathology ; Adult ; Biomarkers, Tumor ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; HSP27 Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Middle Aged ; Neoplasm Invasiveness ; Oncogene Proteins ; genetics ; metabolism ; Pituitary Neoplasms ; metabolism ; pathology ; Protein Deglycase DJ-1 ; Signal Transduction ; Young Adult

Objective To summarize the success of congenital penile curvature. Methods 401 cases of con-genital penile curvature,whose average age is 2. 5 years ,ranging from 1 year to 26 years old. Of all cases ,383 patients who have both hypespadias and penile cun, ature, only 18 patients are congenital penile curvatures. 295 paitents are slight curvature,85 patients are moderate curvature and 21 patients are severe curvature. Curvature correction was per-formed on a case-by-case basis by solution fibric trabs of ventrial urethra and surrounding,skin de-gloving, ff necessa-ry,dorsal plication,and confirmed by Gittes test. Results . Curvature correction was possible by mobilization of ure-thra after penile degloviag,only a few need dorsal plieation. Followup for 6 months to 2 years,396 patients are success at once. The others after a second surgery had 100% success. Conclusions Solution fibrie trabs of ventrial urethra and surrounding, skin de-gloving, if necessary,dorsal plication ,is the first choice to repair of the congenital penile cur-vature and have good results.