Objective To investigate whether baicalein inhibits the proliferation, cell cycle of and pseudopod formation in A431, a skin squamous cell carcinoma cell line, by suppressing the expression of ezrin protein. Methods A431 cells were grouped to be transfected with ezrin-targeting siRNA (siRNA group), treated with baicalein of 5, 10, 20, 40 μmol/L, respectively (baicalein group), or remain untreated (control group). After additional culture, wound healing assay and Transwell assay were performed to observe the migration and invasion of A431 cells, RT-PCR to detect the mRNA expression of ezrin in A431 cells, Western blot and immunoflu-orescence to measure the expression of ezrin protein and its phosphorylation. The pseudopod formation in A431 cells was observed by using scanning electron microscopy. Results After 24-hour culture, wound healing assay displayed that the percent wound closure was 13.3 ± 1.7, 7.6 ±1.6 and 5.9 ± 1.3, respectively, in A431 cells treated with baicalein of 5, 10, 20μmol/L, significantly lower than that in untreated A431 cells (16.3 ± 2.3, all P < 0.01), and the inhibition of baicalein on the migration of A431 cells was concentration-dependent. In the Transwell assay, a significant decrease was observed in the number of A431 cells per high power field permeating through the artificial basement membrane in the groups treated with baicalein of 5, 10, 20 μmol/L for 48 hours compared with the control group (46.5 ± 3.8, 34.3 ± 3.4, 25.3 ± 2.3 vs 56.3 ± 3.8, all P < 0.01), whereas no significant difference was noted between these baicalein-treated groups and siRNA-transfected group (28.3 ± 2.1, all P > 0.05). RT-PCR analysis showed that the mRNA expression of ezrin in baicalein-treated A431 cells significantly decreased compared with that in untreated cells (all P< 0.01), but showed no difference from that in siRNA group (P > 0.05). A statistical difference was also observed in the expression of ezrin and phosphorylated ezrin protein between baicalein-treated A431 cells and untreated cells (all P< 0.05), but not between 40 μmol/L baicalein-treated A431 cells and siRNA-transfected cells (P> 0.05). Furthermore, the suppression of baicalein on ezrin protein and mRNA expression was concentration dependent. The number of pseudopod per cell was significantly lower in 20 μmol/L baicalein-treated A431 cells and siRNA-transfected cells than that in untreated A431 cells (5.3 ± 1.9, 4.5 ± 2.8 vs 22.6 ± 2.8, both P < 0.01), while no significant difference was observed between the former two groups of cells (P > 0.05); the length of pseudopodia also reduced in baicalein-treated cells. Conclusions Baicalein may inhibit the proliferation and invasion of A431 cells by directly or indirectly suppressing the expression of ezrin and phosphorylated ezrin, which in turn contributes to the effect of baicalein against tumor proliferation and metastasis.

blood of patients.

Objective To investigate the expression of ezrin in seborrheie keratosis(SK),basal cell carcinoma (BCC).squamous cell carcinoma (SCC)and its relation to elinical pathology parameters.Methods Skin samples were collected from 36 patients with SCC,27 patients with BCC,20 patients with SK and 10 normal human controls.Immunohistochemistry was performed to detect the expression of ezrin in these samples.The relationship between ezrin expression and prognosis of SCC was assessed using COX regression analysis.Results The positivity rate of ezrin in SK,BCC and SCC samples was 25.0%,66.7%,91.3%,respectively,compared to 20.0%in normal controls.Compared with the controls,a significant increase was observed in the expression of ezrin in patients with BCC and SCC(both P＜0.05).but not in those with SK(P＞0.05).Moreover.increased expression level of ezrin was correlated with a high degree of tumor malignancy,advanced pathological stage,and occurrence of lymph node metastasis of SCC (r=0.87,0.80,0.89,respectively,all P＜0.01).COX regression analysis revealed that the expression level of ezrin was an independent factor for the prognosis of SCC.Conclusion The expression level of ezrin is closely related to the malignancy of skin tumors,pathological grading and lymph node metastasis of SCC.

Objective : To investigate the epidemiological characteristics of heatstroke and its correlation with meteorological factors in Shaoxing in 2017,and to provide evidence for heatstroke prevention and control. Methods : The data of heatstroke cases and the daily meteorological indexes were collected from July 2017 to August 2017 in Shaoxing to describe the spatial,temporal and population distribution of heatstroke cases. The correlation between heatstroke and meteorological factors was analyzed by multivariate logistic regression model. Results : A total of 759 cases of heatstroke were reported,with an average age of（53.3 ±17.9）years. There were 487 males cases（64.16%）and 272 female cases（35.84%）. There were 618 cases of mild heatstroke（81.42%）and 141 cases of severe heatstroke（18.58%）. There were six cases of death from severe heatstroke,and the mortality of severe heatstroke was 4.26%. Minimum temperature（rs=0.851,P<0.001）,maximum temperature（rs=0.726,P<0.001）and wind speed（rs=0.285,P=0.025）were positively correlated with the incidence of heatstroke,and relative humidity（rs=-0.693,P<0.001）and rainfall（rs=-0.414,P=0.001）were negatively correlated with the incidence of heatstroke. The results of logistic regression analysis showed that high daily minimum temperature was a risk factor for severe heatstroke（OR=1.854,95%CI：1.606-2.140）. Conclusion The mortality of severe heatstroke patients was high in Shaoxing,the daily minimum temperature was correlated with severe heatstroke.

Objective Previous study had shown that signal transduction was changed when Hela cells were transfected with HLA-B27gene.An increase of monocyte chemotactic protein-1(MCP-1)of Hela cells was observed.To determine the role of MCP in the pathogenesis of ankylosing spondylitis(AS),expression of MCP-1,2,3and4in peripheral blood mononuclear cells(PBMC),synovial fluid mononuclear cells(SFMC)and synovial tis-sues of AS patients were tested.Methods Gene expression profiles of PBMC from AS patients and healthy volun-teers were determined by cDNA microarray with1176target gene filter.The differentiated expressed gene MCP-1in PBMC,SFMC and synovial tissue of AS patients were confirmed by semi-quantitative RT-PCR.Results The gene expression profile of SFMC of AS patients was significantly different from those of PBMC from AS and PBMC from healthy volunteers.The MCP-1level was positively correlated with MCP-3(r=0.76,P=0.03).The expressions of MCP-1were higher in synovial tissues of AS than those of healthy volunteers(P=0.0035).MCP-1levels in monocytes of AS patients and control subjects were increased after LPS stimulation for4hours.Conclusions There is increased expression of MCP-1in SFMC and synovial tissue of AS patients.The results indicate that MCP-1may play a potential key role in the homing of cells migrating from blood to joint and in the pathogenesis of joint inflammation in AS patients.

Objective To set up a three-dimensional finite element model (FEM) to investigate biomechanics of point contact locking plate (PC-LP) fixating femoral shaft fractures. Methods One intact fresh adult cadaveric femur was scanned by CT at 1 mm interval. Then, the data of CT were utilized to establish three-dimensional FEM by using software Mimics and PRO/E and simulate the different clini-cal loading conditions. The changes of theoretical stress of femur and PC-LP were analyzed under flexion, axial compression and torsion loads. Results (1) Under four-point bending load, the distribution of femur stress was in uniformity, with the largest stress of the PC-LP focused on the edge. (2) Under axial compression load of 250 N, the largest stress of the femur was focused on the screw holes on beth distal ends, with the largest stress of the PC-LP focused on the middle screw holes. (3) Under the torsion load cused on the middlepart and the middle screw holes. Conclusions Under the four-point bending, ax-ial compression and torsion loads, the distribution of femur stress is in uniformity, when the largest stress of the PC-LP focuses on edge or the middle screw holes, while that of the PC-LP on two screw holes of proximal or distal ends.

To explore the role of dentritic epidermal T lymphocytes ( DETCs) in immune rejection of skin allograft in mice and its related mechanism. Methods (1) Full-thickness skin was harvested from back of one male wild type (WT) C57BL/6 mouse. Epithelial cells were isolated for detection of the expression of DETCs and their phenotype with flow cytometer. Another male WT C57BL/6 mouse was used to harvest full-thickness skin from the back. Epidermis was isolated for observation of the morphological characteristics of DETCs with immunofluorescence technology. (2) Four male green fluorescence protein (GFP)-marked C57BL/6 mice, 7 female WT C57BL/6 mice (group WT), and 7 female ybT lymphocytes 8 gene knock-out (GK) C57BL/6 mice (group GK) were used. Full-thickness skin in the size of 1.4 cm x 1.4 cm on the back of mice in groups WT and GK were excised, and the wounds were transplanted with full-thickness skin in the size of 1.2 cm x 1.2 cm obtained from male GFP-marked C57BL/6 mice. The survival time of skin grafts was affirmed with small animal in vivo imager and naked eyes and recorded. (3) Two male WT C57BL/6 mice were used to isolate epithelial cells. Cells were inoculated into 48-well plate and divided into activation group (A) and control group (C) according to the random number table, with 4 wells in each group. Cells in group A were treated with 10 pL concanavalin A in the concentration of 2 microg/mL for 24 hours, while those in group C with PBS in the same volume as that in group A. The expression of interferon y in DETCs was detected with flow cytometer. (4) Four male GFP-marked C57BL/6 mice were used as donors. Fourteen female WT C57BL/6 mice were used as receptors and divided into interferon gamma neutralizing group (IN) and control group (C) according to the random number table, with 7 mice in each group. The skin transplantation model of C57BL/6 male to C57BL/6 female was established as in part (2). Before surgery and 72 hours after, mice in group IN were intraperitoneally injected with 200 pL interferon y neutralizing antibody in the concentration of 1 mg/mL, and those in group C with normal saline in the same volume as that in group IN. The survival time of skin grafts was observed and recorded using the methods in part (2), and the result of group IN was compared with that of group GK in part (2). The survival curve of skin grafts was processed with Log-rank ( Mantel-Cox) test. Results (1) The positive expression rate of DETCs in epithelial cells of skin in mouse was 7.27%, and they were all CD3 cells. DETCs were found to be scattered in the epidermis of skin in mouse with dendritic morphology. (2) The survival time of skin grafts of mice in group GK was 22-35 d, obviously longer than that in group WT (12-16 d, y2 = 14. 10 , P < 0.001). (3) Expression of interferon gamma was detected in 22. 70% DETCs in group A, which was obviously higher than that in group C (0.51%). (4) The survival time of skin grafts of mice in group IN was 19-24 d, which was obviously longer than that in group C (12-16 d, chi 2 = 13.60, P < 0.001) but close to that in group GK as in part (2) (chi2 = 0.06, P = 0.810). Conclusions DETCs are involved in promotion of immune rejection of skin allograft probably by secretinf interferon gamma.
Allografts ; Animals ; Epidermis ; Female ; Graft Survival ; immunology ; physiology ; Interferon-gamma ; immunology ; metabolism ; Lymphocytes ; Male ; Mice ; Mice, Inbred C57BL ; Skin ; Skin Transplantation ; T-Lymphocytes ; immunology

Objective To evaluate the efficacy and safety of two forms of preparations of dexamethasone palmitate in the treatment of rheumatoid arthritis (RA).Methods A multicenter,double-blind,randomized,parallel-group clinical trial was carried out according to good clinical practice (GCP).A total of 237cases of RA patients with mild to moderate knee swelling were randomly divided into the treatment group (n=118 ) or the control group (n=119) and were treated with two kinds of dexamethasone palmitate 8 mg injection respectively.The primary efficacy endpoints were the circumference of the knee joint at the upper and the lower edge after the intra-articular injection.The secondary efficacy endpoints were joint tenderness index and patients general assessment.The adveme events were recorded.Analysis of covariance,t test or Wilcoxon test,x2 test or Fisher exact test were used for statistical analysis.Results The upper edges of the treatment group and the control group after treatment were (37.2±3.3) cm and (36.4±3.9) cm respectively,and the lower edges of the two groups were (34.4±2.9) cm and (33.9±3.4) cm respectively.They were all significantly smaller than the edges before treatment [(38.1± 3.3) cm and (37.3±4.0) cm of the upper edges,(35.1±3.0)cm and (34.6±3.6) cm of the lower edges respectively ) (P＜0.O1)].After treatment,the joint tenderness index were improved (P＜0.01).A total ratio of great improvement and improvement of patients general assessment of the two group patients were 67.5％ (79/117) and 74.8％ (86/115) respectively.No statistical significant difference was found in all primary and secondary efficacy endpoints between the two groups (P＞0.05).During the clinical trial,the incidence of adverse events related to the treatment of two groups were 4.2％ and 6.8％,without any significant difference (P＞0.05).Conclusion New preparation of dexamethasone palmitate has the same efficacy and safety as the imported producted in the treatment of RA.The circumference of the knee joints at the upper and the lower edge may be used to assess the effects of intra-articular injections.

Objective Non-invasive detection is the focus of intense research in diagnosis of acute renal allograft rejection currently. Urine protein is considered the cue to reflect the pathological changes in kidney disease. In this study, we explored the urine markers for early acute renal allograft rejection. Methods The urine protein of two patients with acute renal allograft rejection were examined by 2D gel electrophoresis and bioinformatics. We adopted pH 4-7 ready strip IPG and stained the gel with Sypro-Ruby. The digitized 2D maps of urine protein were quantitatively analyzed using 2D-analysis software packages. By analyzing the differential expressions of proteome between different time points (1, 2, 3 days before acute rejection and 7, 14, 21 days after acute rejection), 30 protein spots were selected and analyzed by MALDI-TOF-MS/MS. Results We obtained 2D gel electrophoresis maps of urine protein of the patients with acute renal allograft rejection, which are of good reproducibility and resolution. Sixteen protein spots were identified, resulting in thirteen corresponding proteins. Out of these proteins, we screened three proteins (alpha-1-antichymotrypsin, tumor rejection antigen gp96, Zn-Alpha-2-Glycoprotein) closely related to acute rejection. Conclusion The urine protein spots on 2D gel electrophoresis maps for the patients with acute renal allograft rejection were of obvious difference when detected at different time points of acute rejection. Alpha-1-antichymotrypsin, tumor rejection antigen gp96 and Zn-Alpha-2-Glycoprotein might be the candidate protein markers to diagnose acute renal allograft rejection after renal transplantation.

The treatment of advanced malignant tumors usually includes chemotherapy, radiotherapy, immunotherapy and targeted therapy, etc. However, a single treatment often faces problems such as drug resistance, toxicity and side effects. Consequently, it is urgent to seek new methods. In 2015, Siwen Hu-Lieskovan proposed "oncology cocktail therapy" firstly. Cocktail therapy is the integration of three or more different methods to exert a synergistic anti-tumor effect. In recent years, many researchers have applied cocktail therapy in basic and clinical researches. For example, the combination of chemotherapy drugs, targeted drugs and immunotherapy drugs makes the advantages of drugs complementary. The combination of hyperthermia, chemotherapy and immunotherapy can improve the tumor immunosuppressive microenvironment and stimulate immunity, and then combines with anti-programmed death receptor 1 (aPD-1) drugs to produce synergistic effects. The results show that cocktail therapy can improve the efficacy and prognosis of advanced cancer patients. In this article, the role and impact of cocktail therapy were reviewed, aiming to provide reference for in-depth exploration of new combined treatments for advanced malignant tumors.