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.

Objective:To explore the repairing effect and mechanism of tangeretin in rats with spinal cord injury.Methods:The rats were divided into sham-operation group, model group and tangeretin group according to random number table, with 8 rats in each group. Except for the sham-operation group, Allen hit method was used to make rat models in the other groups. After the model was successfully established, the tangeretin group was intragastrically administered with tangeretin 50 mg/kg, and the sham-operation group and the model group were intragastrically administered with an equal volume of normal saline once a day for 14 days. On days 0, 3, 7, and 14 after modeling, the motor function recovery of rats was assessed using the Basso-Beattie-Bresnahan (BBB) score; the morphological changes of the spinal cord tissues were observed using HE staining and Nissl staining; the SOD and GSH activities and MDA, IL-1β, TNF-α, and IL-10 levels in the spinal cord tissues of rats in each group were measured using ELISA kit detection; the GFAP and Neun expressions in the spinal cord tissues were detected by immunofluorescence; the IL-1β, TNF-α, IL-10, nuclear factor E2-related factor 2 (Nrf-2), and NAD (P) H-quinone oxidoreductase 1 (NQO-1) expressions in the spinal cord tissues were detected by Western blot.Results:Compared with the model group, the BBB score increased in the tangeretin group ( P<0.05), HE staining score decreased ( P<0.05), and the number of Nissl bodies increased ( P<0.05); the level of IL-10, SOD and GSH activities increased ( P<0.05), and IL-1β, TNF-α and MDA levels decreased in the spinal cord tissue ( P<0.05); GFAP fluorescence intensity decreased ( P<0.05) and NeuN fluorescence intensity increased ( P<0.05); the relative expression of IL-1β and TNF-α decreased ( P<0.05), and the relative expressions of IL-10, Nrf-2 and NQO-1 protein increased ( P<0.05). Conclusions:Tangeretin can exert anti-inflammatory and anti-oxidative stress effects through the Nrf2/NQO1 signaling pathway and alleviate early spinal cord injury in rats. On the other hand, it may promote the recovery of spinal cord injury by reducing glial scar generation and promoting neural cellogenesis.

OBJECTIVES: Malignant melanoma is a highly malignant and heterogeneous skin cancer. Although immunotherapy has improved survival rates, the inhibitory effect of tumor microenvironment has weakened its efficacy. To improve survival and treatment strategies, we need to develop immune-related prognostic models. Based on the analysis of the Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), and Sequence Read Archive (SRA) database, this study aims to establish an immune-related prognosis prediction model, and to evaluate the tumor immune microenvironment by risk score to guide immunotherapy. METHODS: Skin cutaneous melanoma (SKCM) transcriptome sequencing data and corresponding clinical information were obtained from the TCGA database, differentially expressed genes were analyzed, and prognostic models were developed using univariate Cox regression, the LASSO method, and stepwise regression. Differentially expressed genes in prognostic models confirmed by real-time reverse transcription PCR (real-time RT-PCR) and Western blotting. Survival analysis was performed by using the Kaplan-Meier method, and the effect of the model was evaluated by time-dependent receiver operating characteristic curve as well as multivariate Cox regression, and the prognostic model was validated by 2 GEO melanoma datasets. Furthermore, correlations between risk score and immune cell infiltration, Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data (ESTIMATE) score, immune checkpoint mRNA expression levels, tumor immune cycle, or tumor immune micro-environmental pathways were analyzed. Finally, we performed association analysis for risk score and the efficacy of immunotherapy. RESULTS: We identified 4 genes that were differentially expressed in TCGA-SKCM datasets, which were mainly associated with the tumor immune microenvironment. A prognostic model was also established based on 4 genes. Among 4 genes, the mRNA and protein levels of killer cell lectin like receptor D1 (KLRD1), leukemia inhibitory factor (LIF), and cellular retinoic acid binding protein 2 (CRABP2) genes in melanoma tissues differed significantly from those in normal skin (all P<0.01). The prognostic model was a good predictor of prognosis for patients with SKCM. The patients with high-risk scores had significantly shorter overall survival than those with low-risk scores, and consistent results were achieved in the training cohort and multiple validation cohorts (P<0.001). The risk score was strongly associated with immune cell infiltration, ESTIMATE score, immune checkpoint mRNA expression levels, tumor immune cycle, and tumor immune microenvironmental pathways (P<0.001). The correlation analysis showed that patients with the high-risk scores were in an inhibitory immune microenvironment based on the prognostic model (P<0.01). CONCLUSIONS The immune-related SKCM prognostic model constructed in this study can effectively predict the prognosis of SKCM patients. Considering its close correlation to the tumor immune microenvironment, the model has some reference value for clinical immunotherapy of SKCM.
Humans ; Melanoma/genetics* ; Skin Neoplasms/genetics* ; Tumor Microenvironment ; Prognosis

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: To evaluate effects of acitretin and methotrexate on treatment of severe plaque psoriasis.
 METHODS: A total of 54 patients were treated with either acitretin (32 patients; 30 mg per day) or methotrexate (22 patients; 15 mg per week) for 8 weeks. The therapeutic effects of the two drugs were evaluated according to the psoriasis area and severity index (PASI).
 RESULTS: After 8 weeks of treatment, the score of PASI decreased in both the acitretin group and the methotrexate group (P<0.001). However, there was no significant differences neither in the percentage reduction in the PASI score nor in PASI response rate between the two groups (P>0.05).
 CONCLUSION Acitretin and methotrexate exert similar therapeutic effect in severe plaque psoriasis.
Acitretin ; Humans ; Methotrexate ; Psoriasis

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 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.