Cancer immunotherapy has become a promising strategy. However, the effectiveness of immunotherapy is restricted in "cold tumors" characterized with insufficient T cells intratumoral infiltration and failed T cells priming. Herein, an on-demand integrated nano-engager (JOT-Lip) was developed to convert cold tumors to hot via "increased DNA damage and dual immune checkpoint inhibition" strategy. JOT-Lip was engineered by co-loading oxaliplatin (Oxa) and JQ1 into liposomes with T-cell immunoglobulin mucin-3 antibodies (Tim-3 mAb) coupled on the liposomal surface by metalloproteinase-2 (MMP-2)-sensitive linker. JQ1 inhibited DNA repair to increase DNA damage and immunogenic cell death (ICD) of Oxa, thus promoting T cells intratumoral infiltration. In addition, JQ1 inhibited PD-1/PD-L1 pathway, achieving dual immune checkpoint inhibition combining with Tim-3 mAb, thus effectively promoting T cells priming. It is demonstrated that JOT-Lip not only increased DNA damage and promoted the release of damage-associated molecular patterns (DAMPs), but also enhanced T cells intratumoral infiltration and promoted T cell priming, which successfully converted cold tumors to hot and showed significant anti-tumor and anti-metastasis effects. Collectively, our study provides a rational design of an effective combination regimen and an ideal co-delivery system to convert cold tumors to hot, which holds great potential in clinical cancer chemoimmunotherapy.

The present study was conducted to clarify the therapeutic effect of cornuside on experimental autoimmune encephalomyelitis (EAE) and its influence on T helper 17 (Th17) cell and regulatory T (Treg) cell infiltration into the central nervous system. Rats were randomly placed into four treatment groups: control, EAE, EAE+cornuside, and EAE+prednisolone. The neurological function scores of rats were assessed daily. On the second day after EAE rats began to show neurological deficit symptoms, the four groups were treated with normal saline, normal saline, cornuside (150 mg/kg), and prednisolone (5 mg/kg), respectively. The treatment was discontinued after two weeks, and the spinal cord was obtained for hematoxylin and eosin (H&E) and luxol fast blue staining, as well as retinoic acid receptor-related orphan receptor γ (RORγ) and forkhead box protein P3 (Foxp3) immunohistochemical staining. Blood was collected for Th17 and Treg cell flow cytometry testing, and the serum levels of interleukin (IL)-17A, IL-10, transforming growth factor-β (TGF-β), IL-6, IL-23, and IL-2 were measured via enzyme-linked immunosorbent assay (ELISA). Compared with rats in the EAE group, rats in the EAE+cornuside and EAE+prednisolone groups began to recover from neurological deficits earlier, and had a greater degree of improvement of symptoms. Focal inflammation, demyelination, and RORγ-positive cell infiltration were reduced by cornuside or prednisolone treatment, whereas the Foxp3-positive cell numbers were not significantly different. Meanwhile, the number of Th17 cells and the IL-17A, IL-6, and IL-23 levels were lower in the blood after cornuside or prednisolone treatment, whereas the number of Treg cells or the levels of IL-10, TGF-β, and IL-2 were not markedly different. Cornuside can alleviate symptoms of EAE neurological deficits through its anti-inflammatory and immunosuppressive effects, and Th17 cells may be one of its therapeutic targets.

Objective:To discuss the application and effectiveness of the flipped classroom based on Wechat platform in the teaching of neurology.Methods:clinical students of grade 2014 of binjiang college of Zhejiang Chinese Medical University were selected as teaching subject. The students were divided into experimental group which contained a total of 74 students in class one and class three and the control group which contained a total of 38 students in class two. Peripheral neuropathy and neuromuscular-junction disease were selected as teaching contents. The experimental group adopted flipped classroom as the teaching mode and Wechat platform for pre-class preparation, after-school review and interactive communication. The control group was taught by traditional teaching mode. Before and after class, students in the two groups had a small-scale test and were surveyed by questionnaire, respectively. Students in the two groups took the same final exam which included case analysis when the course was over. The scores of the final exam and the results of the case analysis of each group were recorded and analyzed. All data were processed with statistical software SPSS 20.0, and t-test or chi-square test was used. Results:Students in the experimental group had significantly higher test scores in the after-class small-scale test than those in the control group ( P=0.038). Their final exam scores were higher than those in the control group ( P=0.046), and their scores of case analysis in the final exam were higher than those in the control group ( P=0.026). The results of pre-class questionnaire survey showed that the proportion of students who chose "good" in the experimental group was higher than that in the control group on the understanding of the learning content and the preparation ( P<0.05). In the after-class questionnaire survey, the proportion of students who chose "excellent" and "good" in the evaluation of learning interest in the experimental group was higher than those in the control group ( P<0.05), the proportion of students in the experimental chose "good" in evaluating their self-learning ability was higher than that in the control group ( P<0.05), the proportion of "excellent" on clinical thinking ability and teaching satisfaction was higher in the experimental group than that in the control group ( P<0.05), and the overall proportion of students who chose "excellent" and "good" in all items in the experimental group were higher than that in the control group ( P<0.05). In the experimental group, the overall proportion of students who selected "excellent" and "good" in on the evaluation of their learning interest was significantly increased in the after-class questionnaire survey compared with the pre-class questionnaire survey ( P<0.01). Conclusions:The application of flipped classroom based on Wechat platform is feasible and effective in the teaching of neurology. It can make up for the deficiency of traditional teaching methods. It is helpful to improve students' learning interest and self-learning ability, and is also helpful to exercise their clinical thinking ability. Thus this method deserves further popularization.

PURPOSE: The intermediate stage of hepatocellular carcinoma (HCC) (Barcelona Clinic Liver Cancer [BCLC] B) comprises a highly heterogeneous population, and the treatment strategy is still controversial. Because of the heterogeneity, a subclassification of intermediate-stage HCCs was put forward by Bolondi according to the ‘beyond Milan and within up-to-7' criteria and Child-Pugh score. In this study, we aim to analyze the prognosis of BCLC-B stage HCC patients who received hepatic resection according to the Bolondi's subclassification. MATERIALS AND METHODS: One thousand and one hundred three patients diagnosedwith HCC and treatedwith hepatic resectionwere enrolled in our hospital between 2006 and 2012. According to Bolondi's subclassification, the BCLC-B patients were divided into four groups. Recurrence-free survival (RFS) and overall survival (OS) were analyzed. RESULTS: According to Bolondi's subclassification, the BCLC-B patients were divided into four groups: B1 (n=41, 18.7%), B2 (n=160, 73.1%), B3 (n=11, 5.0%), and B4 (n=7, 3.2%). Significant difference was observed between B1 and other groups (B1 vs. B2, p=0.022; B1 vs. B3, p < 0.001; B1 vs. B4, p < 0.001), but no difference for B2 vs. B4 (p=0.542) and B3 vs. B4 (p=0.542). In addition, no significant differences were observed between BCLC-A and BCLC-B1 group for both RFS (p=0.087) and OS (p=0.643). In multivariate analysis, BCLC-B subclassification was not a risk factor for both OS (p=0.263) and RFS (p=0.892). CONCLUSION: In our study, HCC patients at B1 stagewere benefited from hepatic resection and had similar survival to BCLC-A stage patients. Our study provided rationality of hepatic resection for selected BCLC-B stage HCC patients instead of routine transarterial chemoembolization.
Carcinoma, Hepatocellular* ; Hepatectomy ; Humans ; Liver Neoplasms ; Multivariate Analysis ; Population Characteristics ; Prognosis ; Risk Factors

Objective To explore the CRP harmonization by calibration using commutable trueness verification materials. Methods High and low level of CRP concentrations trueness verification materials(H and L) were prepared by Beijing center for clinical laboratories. Thesetrueness verification materials were diluted to 5 calibration points(5L, 4L+1H, 3L+2H, 1L+4H, 5H) by weighing method, respectively. These 5 points were used to calibrate four different brands of CRP detection system (Diasys, Leadman, Siemens and Roche) instead of the original procedure. Sera from 21 patients and the international standard ERM DA-474/IFCC were used to compare harmonization and trueness after calibration. Each sample above was measured twice. Results After calibration, the median of CV was reduced from 19.33％ to 2.92％ among 21 patient samples, less than the optimal CV based on biological variability (CV=10.6％). Compared with Desai, the slopes were closer to 1 from 0.90-1.09 to 0.93-0.96 after calibration. Meanwhile, if ERM-DA474/IFCC was used as the trueness verification materials, the absolute bias wasreduced from 3.08-11.07 mg/L to 0.52-2.97 mg/L which was close to theuncertainty of itself (2.5 mg/L). Conclusions Afterthe calibration which contained five linear concentration points of CRP trueness verification materials by weighing method, both harmonization and trueness of CRP were improved.

Objective To prepare the trueness verification materials of C-reactive protein (CRP) and evaluate its homogeneity, stability and commutability. Methods The high and low CRP concentrations trueness verification materials were from patient leftover sera which were pooled, mixed thoroughly, filtered and aliquoted. The homogeneity, stability and commutability of these materials were evaluated according to CNAS(China National Accreditation Service for Conformity Assessment, CNAS)-GL29:2010 "Reference materials-General and statistical principles for certification (ISO Guide35:2006)"and the Clinical and Laboratory Standards Institute (CLSI) EP30A. The trueness verification materials were used to evaluate the commutability in 10 clinical CRP detection systems, using forty-five patients' leftover sera with different CRP concentration evaluated by Deming regression in EP30A of CLSI. Meanwhile, the commutability of dilution series of ERM DA-474/IFCC were evaluated using the same method. Results A total of two CRP concentration level trueness verification materials were prepared, with high and low concentration levels of 754 and 743 vials, 1 ml each, respectively. The preparation showed good homogeneity (F

Objective To value C-reactive protein ( CRP ) trueness verification materials and to perform the CRP trueness verification program in Beijing .Methods The CRP value of trueness verification materials were assigned by the international reference material ERM DA-474/IFCC, using 10 clinical routine detection systems at departments of clinical laboratory of Beijing Chaoyang and Luhe Hospital Affiliated to Capital Medical University .The calibration curves with 4 ERM DA-474/IFCC dilutions were established and used for value transfer for trueness verification materials of two levels .The uncertainty was also assessed during the process.Then, the trueness verification was performed in the EQA at Beijing Center for Clinical Laboratories ( BCCL ) among 42 clinical laboratories.The samples were distributed according to BCCL standard operating procedure .The Microsoft Excel 2007 and SPSS 17.0 were used to process the results and the function of efficiency ( En) was calculated to verify the difference between the value and the overall mean of all participating laboratories .Results The values and uncertainties of two trueness verification materials of CRP were (109.9 ±9.4) mg/L and (27.1 ±2.4) mg/L respectively.The results of trial application of two level trueness verification materials in the EQA at Beijing Center for Clinical Laboratories (BCCL) were satisfied.There were no significant difference between the transfer values from our study and the values from means of all laboratories in Beijing .The function of efficiency ( En ) was less than 1.Conclusions The valueswhich were established by using multiple detection platforms for CRP trueness verification materialswere accurate and the uncertainties were small .This method is a preferably method for CRP value assignment because there was no suitable reference method for CRP measurement till now .Thematerialswere suitable for the trueness verification program for clinical laboratories in Beijing .

OBJECTIVE: To investigate the effect of curcumin on dopamine neurons in Parkinson's disease (PD) and its mechanism. METHODS: SH-SY5Y human neuroblastoma cells were treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to establish the PD cell model. The model cells were treated with curcumin and/or autophagy inhibitor 3-MA. After 48 h of drug treatment, the number of surviving dopamine neurons was detected by tyrosine hydroxylase immunofluorescence method. Western blotting was used to detect protein expression of α-Synuclein (α-Syn), transcription factor EB (TFEB) and autophagy-related proteins lysosome-associated membrane protein 2A (LAMP2A) and microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ); RT-PCR was used to detect mRNA expression of α-Syn. RESULTS: Compared with MPTP model group, curcumin increased the number of surviving dopamine neurons(<0.01), decreased both protein expression and mRNA expression of α-Syn (all <0.01), and increased protein expression of TFEB, LAMP2A and LC3-Ⅱ (all <0.01). When curcumin and 3-MA were given concurrently, the number of surviving dopamine neurons, protein expression of TFEB, LAMP2A and LC3-Ⅱ increased (<0.05 or <0.01), and both protein expression and mRNA expression of α-Syn decreased (<0.05 or <0.01) compared with MPTP model group; but the number of surviving dopamine neurons and protein expression of LAMP2A and LC3-Ⅱ decreased compared with curcumin group (all <0.05). CONCLUSIONS Curcumin exerts protective effect on dopamine neurons in PD, which may be associated with enhancing autophagy and promoting the clearance of α-Syn.
Animals ; Cell Line ; Curcumin ; pharmacology ; Dopaminergic Neurons ; drug effects ; Humans ; Mice ; Mice, Inbred C57BL ; Parkinson Disease ; alpha-Synuclein ; metabolism

Objective To systematically review the safety and efficacy of enhanced recovery after surgery (ERAS) in perioperative management of pancreaticoduodenectomy.Methods A search was performed in databases (including PubMed,EMASE,Cochrane library,Sinomed,Wangfang,VIP,and CNKI) for randomized controlled trials (RCTs) and non-randomized controlled trials (NRCTs) up to September 2016 on use of ERAS in patients undergoing pancreaticoduodenectomy.After quality evaluation and data extraction,meta-analysis was performed using RevMan 5.3.Results Four RCTs and Twelve NRCTs involving a total of 2 828 patients were included.1 401 patients were in the ERAS group,and 1 427 in the control group.Meta-analysis results showed that compared with the control group,the ERAS group had shorter length of hospital stay (SMD =-0.36,95 ％ CI =-0.44--0.28,P＜ 0.05) and lower incidence of delayed gastric emptying (RR =0.61,95％ CI=0.51-0.73,P＜0.05).However,no significant differences were observed in pancreatic fistula rate,bile fistula rate,readmission rate,reoperation rate,and overall mortality morbidity rate between the two groups (all P＞0.05).Conclusion It is reasonably safe and efficacious to adopt ERAS in periopetative management of patients undergoing pancreaticoduodenectomy.

Objective To investigate whether curcumin protects dopamine neurons in Parkinson's disease (PD) by activating autophagy and discuss its specific mechanism.Methods In vitro,SH-SY5Y human neuroblastoma cells were treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPP+) to induce PD cell models.In vivo,C57BL/6 male mice were performed intraperitoneal injection of 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) once daily for 7 d continuously to establish PD models.SH-SY5Y human neuroblastoma cells and C57BL/6 male mice were,respectively,divided into normal control group,model group,curcumin group,curcumin and 3-methyladenine (3-MA) group,and 3-MA group.Curcumin was used as intervention treatment,and 3-MA,an autophagy inhibitor,was used individually or concurrently with curcumin as intervention controls:curcumin (40 μmol/L),curcumin (40 μmol/L)+3-MA (4 mmol/L),and 3-MA (4 mmol/L) were added into the cells of curcumin group,curcumin and 3-MA group,and 3-MA group at the time of MPP+ addition;curcumin (80 mg/kg),curcumin (80 mg/kg)+3-MA (2 mg/kg),and 3-MA (2 mg/kg) were added into the mice of curcumin group,curcumin and 3-MA group,and 3-MA group at the time of MPTP injection once daily.In the normal control group,sterile saline was injected into the abdomen simultaneously (30 rmL/kg).In cell groups,after 48 h of drug treatment,the number of surviving dopamine neurons was detected by tyrosine hydroxylase (TH) immunofluorescence;Western blotting was used to detect protein expressions of α-synuclein (α-Syn),and autophagy related proteins lysosome-associated membrane protein 2A (LAMP2A) and microtubule-associated protein 1 light chain 3 (LC3);transcription factor EB (TFEB) levels in the cytoplasm and nucleus were detected separately by Western blotting;reverse transcription (RT)-PCR was used to detect the α-Syn mRNA expression.In mice groups,14 d after the last injection,mice were sacrificed and sections ofmidbrain nigra were gained for detection of TH level by immunohistochemistry,expressions of α-Syn and LC3 were detected by Western blotting.Results (1) As compared with model group,curcumin group had significantly larger number of surviving dopamine neurons both in vitro and in vivo,significantly decreased α-Syn protein and mRNA expressions in vitro,and significantly decreased α-Syn protein expression in vivo (P＜0.05);curcumin group had statistically increased LAMP2A and LC3-Ⅱ protein expressions in vitro and LC3-Ⅱ protein expression in vivo as compared with model group (P＜0.05).(2) As compared with curcumin group,curcumin and 3-MA group had statistically smaller number of surviving dopamine neurons in vitro and in vivo,and statistically increased α-Syn protein expression in vitro (P＜0.05).(3) In vitro,as compared with model group,curcumin group had significantly increased TFEB protein expression in nucleus (P＜0.05),and curcumin and 3-MA group had statistically increased TFEB protein expression in cytoplasm and statistically decreased TFEB protein expression in nucleus (P＜0.05).Conclusion Curcumin has protective effect on dopamine neurons of PD;one of the main mechanisms is that curcumin can activate the autophagy function and promote the clearance of α-Syn;increasing the TFEB expression,promoting the transfer of TFEB to nucleus to perform the transcription function,and then promoting the synthesis of autolysosome may be the main ways to activate the autophagy function.