Glioblastoma(GBM)is a lethal cancer with limited therapeutic options.Dendritic cell(DC)-based cancer vaccines provide a promising approach for GBM treatment.Clinical studies suggest that other immu-notherapeutic agents may be combined with DC vaccines to further enhance antitumor activity.Here,we report a GBM case with combination immunotherapy consisting of DC vaccines,anti-programmed death-1(anti-PD-1)and poly I:C as well as the chemotherapeutic agent cyclophosphamide that was integrated with standard chemoradiation therapy,and the patient remained disease-free for 69 months.The patient received DC vaccines loaded with multiple forms of tumor antigens,including mRNA-tumor associated antigens(TAA),mRNA-neoantigens,and hypochlorous acid(HOCl)-oxidized tumor lysates.Furthermore,mRNA-TAAAs were modified with a novel TriVac technology that fuses TAAs with a destabilization domain and inserts TAAs into full-length lysosomal associated membrane protein-1 to enhance major histo-compatibility complex(MHC)class Ⅰ and Ⅱ antigen presentation.The treatment consisted of 42 DC cancer vaccine infusions,26 anti-PD-1 antibody nivolumab administrations and 126 poly I:C injections for DC infusions.The patient also received 28 doses of cyclophosphamide for depletion of regulatory T cells.No immunotherapy-related adverse events were observed during the treatment.Robust antitumor CD4+and CD8+T-cell responses were detected.The patient remains free of disease progression.This is the first case report on the combination of the above three agents to treat glioblastoma patients.Our results suggest that integrated combination immunotherapy is safe and feasible for long-term treatment in this patient.A large-scale trial to validate these findings is warranted.

Mice ; Animals ; Alzheimer Disease/complications* ; Entorhinal Cortex/metabolism* ; Brain-Derived Neurotrophic Factor/metabolism* ; Memory Disorders/etiology* ; Thalamus/metabolism* ; Disease Models, Animal

Objective Reports are relatively few at home and abroad onrobot-assistedlaparoscopic ureteral reimplantation ( RAUR) .This study summarizes our experience with RAUR and assesses the feasibility and clinical effect of the strategy . Methods We retrospectively analyzed the clinical data about 5 cases of lower ureterostenosistreated by RAUR in our department from May 2015 to February 2017. Results Operations were successfully completed in all the cases , with an estimatedintraoperative blood loss of 40-100 mL but no blood transfusion , nor conversion to open surgery , nor intestinal or major vascular injury .The urethral catheter was re-moved at 5-7 days after surgery and there were no postoperative complications .The patients were followed up for over 6 months , during which all showed increased glomerular filtration rate ( 2-12 mL/min) of the involved kidney at 3 months and ultrasonography revealed no ureterostenosisorhydronephrosis . Conclusion Robot-assistedlaparoscopic ureteral reimplantation is a safe , effective and minimally invasive surgical option for distal ureteral obstruction .

OBJECTIVETo investigate the impact of interferon-stimulated exonuclease 20 kDa (ISG20) on replication of genotype 2a hepatitis C virus (HCV) subgenomic replicon RNA and infectivity of the cell culture-derived HCV strain JFH1 to determine the potential of exogenously expressed ISG20 as an anti-viral therapy of chronic hepatitis C.

METHODSPlasma vectors containing wild-type (WT) ISG20 or a catalytically-inactive mutant ISG20m were transiently transfected into Huh7, Huh7.5 and HEK293 cells, and the replication of a monocistronic subgenomic JFH1 RNA replicon, SGRm-JFH1BlaRL, was measured. Huh7.5 cells stably expressing ISG20, ISG20m, or the control vector were established by transducing replication incompetent pCX4-Bsr-myc retroviruses encoding WT ISG20, D94G mutant ISG20, or the empty vector, respectively, and selecting with 5 mug/mL of blasticidin for approximately three weeks. The stable Huh7.5 cells were then transfected with HCV replicon RNA and infected with cell culture-derived HCV to investigate inhibition capacity of ISG20 against HCV.

RESULTSHuh7.5-ISG20, Huh7.5-ISG20m, and Huh7.5-Bsr controls cells stably expressing ISG20, ISG20m, or the control vector, respectively, were constructed successfully; the ectopically expressed ISG20 and ISG20m were distributed in both nucleus and cytoplasm, as detected by immuno uorescence. SGRm-JFH1BlaRL replicated efficiently and with similar kinetics in the Huh7.5-Bsr and Huh7.5-ISG20m cells, with expression levels plateauing at 48-96 h post-transfection. In contrast, at all time points examined, SGRm-JFH1BlaRL replication was 9.1% to 16.7% in the Huh7.5-ISG20 cells. The Huh7, Huh7.5 and HEK293 cells transiently expressing ISG20 also showed 16.7% to 25.0% of HCV replication that the respective controls. In addition, the amount of infectious progeny JFH1 virus released in culture supernatants was 9.1% to 12.5% from the Huh7.5-ISG20 cells than from the Huh7.5-Bsr and Huh7.5-ISG20m cells at 48-72 h post-infection, and the latter two cultures produced similar JFH1 virus yields. Finally, the expression of HCV core protein was also lower in the Huh7.5-ISG20 cells, as detected by immunoblot analysis.

CONCLUSIONExogenous expression of ISG20, either in a transient or stable manner, suppresses not only replication of genotype 2a HCV RNA replicons but also JFH1 virus propagation in cultured hepatocytes. The exonuclease activity of ISG20 is required for its antiviral activities against HCV.

Antiviral Agents ; pharmacology ; Cell Line ; Genome, Viral ; HEK293 Cells ; Hepacivirus ; genetics ; Humans ; RNA, Viral ; genetics ; Replicon ; Virus Replication ; drug effects

To construct a hepatitis C virus (HCV) genotype 2a monocistronic replicon and investigate its replication capabilities in the human hepatocarcinoma cell lines, Huh7.5 and Huh7.1, in order to determine its potential as a molecular tool for future in vitro studies of HCV replication and selection studies for putative anti-HCV drugs. Site-directed mutagenesis was used to delete the Core-E1-E2-p7-NS2 fragment (about 3090 bp) from plasmid pJ6JFH1BlaRL. The resultant trianglepJ6JFH1BlaRL plasmid was digested with AgeI and AvrII to release the cDNA fragment (hereafter, referred to as fragment L) containing partial 5'-untranslated region (UTR), the first 12 amino acid (aa) of HCV Core coding sequence, full-length coding sequences for the blasticidin-resistance gene, Renilla luciferase, foot-and-mouth disease virus (FMDV) 2a antiprotease and ubiquitin, and partial coding sequence for HCV NS3. To generate the monocistronic replicon, pSGRmJFH1BlaRL, fragment L was ligated into the pSGR-JFH1 vector that had been digested with AgeI and AvrII to remove the partial 5'-UTR, the first 19 aa of HCV Core coding sequence, the full-length coding sequence for the neomycin phosphotransferase II gene, the internal ribosomal entry site from encephalomyocarditis virus, and partial HCV NS3 coding sequence. A replication-defective mutant replicon, pSGRmJFH1BlaRL/GND, was constructed by a similar procedure using the pSGR-JFH1/GND vector. Fragment L was confirmed in both constructs by sequencing. Replicon RNAs were prepared from XbaI-linearized plasmid DNA templates with Invitrogen's T7 MEGAscript kit, and were purified by DNase I treatment and LiCl precipitation. RNAs were quanti?ed by optical density, and the quality and concentration were con?rmed by agarose gel electrophoresis. Replicon RNAs were transfected into Huh7.5 and Huh7.1 cells using Invitrogen's DMRIE-C transfection reagent at a ratio of 5 mug of lipid to 1mug of RNA. Time course assay of Renilla luciferase activity indicated the replicon's replication function. The pSGRmJFH1BlaRL monocistronic replicon and pSGRmJFH1BlaRL/GND replication-defective mutant replicon were successfully constructed. The pSGRmJFH1BlaRL replicon was replication-proficient in Huh7.5 and Huh7.1 cells, with replication peaking at 72 hours post-transfection and decreasing after 96 hours. No replication was detected at any time point post-transfection for the defective mutant replicon. A monocistronic replicon of HCV genotype 2a was constructed and shown to be replication-proficient in human hepatocarcinoma cell lines.
Cell Line, Tumor ; Genetic Vectors ; Genome, Viral ; Genotype ; Hepacivirus ; genetics ; Humans ; Mutagenesis, Site-Directed ; RNA, Viral ; Transfection ; Virus Replication

BACKGROUNDChromosomal abnormalities have been shown to play an important prognostic role in multiple myeloma (MM). Interphase fluorescence in situ hybridization (i-FISH) has been much more effective to identify cytogenetic aberrations in MM than conventional cytogenetic technique (CC). To clearly determine the cytogenetic features of Chinese MM patients and identify their prognostic implications, we designed a multicenter study based on i-FISH including 672 patients from 52 hospitals in China.

METHODSAll 672 patients were systematically screened for the following genomic aberrations: del(13q), IgH rearrangement, del(p53) and 1q21 amplifications.

RESULTSThe analysis showed that the chromosomal changes were detected in 22.1% patients by CC and in 82.3% patients by i-FISH. The most common abnormalities by CC were chromosome 1 aberrations (48.4%), -13/13q- (37.6%), hyperdiploidy (36.6%), hypodiploidy (30.1%) and IgH rearrangements (23.7%). The most frequent abnormalities by FISH was del(13q), which was found in 60.4% patients, whereas IgH rearrangement, 1q21 amplification and p53 deletions were detected in 57.6%, 49.0% and 34.7% cases, respectively. By statistical analysis, -13/13q- by CC was associated with low level of platelet (P = 0.015), hyperdiploidy was associated with low level of serum albumin (P = 0.028), and IgH rearrangement by FISH was associated with high level of β2 microglobulin (P = 0.019). Moreover, 1q21 amplification and del(p53) by FISH conferred a high incidence of progressive disease (PD) after initial therapy. Metaphase detection of IgH rearrangements and chromosome 1 aberrations concurrently was associated with a short progression free survival (PFS) (P = 0.036). No significant prognostic implications of other cytogenetic abnormalities were found associated with overall survival and PFS.

CONCLUSIONSChinese MM patients had similar cytogenetic abnormalities compared with the previous reported studies. However, the prognostic significance of FISH aberrations were not clearly determined and further study is required.

Adult ; China ; Chromosome Aberrations ; Chromosomes, Human, Pair 1 ; genetics ; Cytogenetic Analysis ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology

BACKGROUNDThree randomised trials have demonstrated that combining bevacizumab with first-line chemotherapy significantly improves progression-free survival versus chemotherapy alone in HER2-negative locally recurrent/metastatic breast cancer (LR/mBC). However, data from Chinese populations are limited and possible differences between ethnic and geographic populations are unknown. This study was conducted to determine whether there are differences in safety and efficacy in patients with HER2-negative LR/mRC between Chinese and Western populations after they receive first-line bevacizumab combined with taxane-based therapy.

METHODSIn the single-arm, open-label, Avastin Therapy for Advanced Breast Cancer (ATHENA) study (NCT00448591), patients with HER2-negative LR/mBC received first-line bevacizumab (investigator's choice of 10 mg/kg every 2 weeks or 15 mg/kg every 3 weeks) combined with taxane-based therapy. The primary endpoint was safety profile and the secondary is time to progression (TTP). A subpopulation analysis was conducted to assess safety and efficacy in Chinese patients.

RESULTSOf 2264 patients treated in ATHENA, 202 were enrolled in China. Bevacizumab was combined with docetaxel in 90% of Chinese patients and paclitaxel in 10%. The most common grade 3/4 adverse events were diarrhoea (in 5.0% of patients) and hypertension (in 2.5% of patients). Grade 3/4 proteinuria occurred in 0.5%. After median follow-up of 17.6 months and events in 56% of patients, median TTP was 9.0 months (95%CI, 8.4-11.1). Overall survival data were immature.

CONCLUSIONSWe found no evidence of increased bevacizumab-related toxicity or reduced efficacy in Chinese LR/mBC patients receiving first-line bevacizumab-taxane therapy compared with predominantly Western populations. The safety profile was generally similar to previously reported LR/mBC trials. Subtle differences may be attributable to different lifestyle and cardiovascular risk factors in Chinese patients compared with the overall population. It appears reasonable to extrapolate findings from bevacizumab-based randomised trials to Chinese populations.

Adult ; Aged ; Aged, 80 and over ; Antibodies, Monoclonal, Humanized ; adverse effects ; therapeutic use ; Antineoplastic Agents ; adverse effects ; therapeutic use ; Bevacizumab ; Breast Neoplasms ; drug therapy ; genetics ; metabolism ; Bridged-Ring Compounds ; adverse effects ; therapeutic use ; Female ; Humans ; Middle Aged ; Receptor, ErbB-2 ; genetics ; metabolism ; Taxoids ; adverse effects ; therapeutic use ; Young Adult

OBJECTIVETo analyze the correlations between increased spinal cord signal intensity on magnetic resonance images (MRI) and the clinical prognosis of compressive cervical myelopathy.

METHODSSixty-six patients with cervical spondylotic myelopathy underwent surgeries through the anterior approach. In all the patients, the diagnoses were established on the basis of both neurological examination and MRI findings that showed spinal cord compression. The patients were divided into two groups according to preoperative MRI, namely isointense MRI T1/T2 signal group and iso/hyperintense MRI T1/T2 group. The JOA scores of the patients were evaluated before and at 6 and 12 months after the operation.

RESULTSThe patients were followed up for 12 to 38 months after the operation (mean 21 months), and no statistically significant difference were found in the pre- and postoperative JOA scores between the two groups (P>0.05).

CONCLUSIONThe peoperative hyperintense signals on T2 weighted MRI does not correlate to the prognosis of patients with compressive cervical myelopathy, who may also have favorable clinical outcomes after the operation.

Adult ; Cervical Vertebrae ; pathology ; surgery ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Prognosis ; Spinal Cord Compression ; diagnosis ; etiology ; surgery ; Spinal Osteophytosis ; complications ; diagnosis ; surgery

Objective To investigate the status of endemic fluorosis in Boxing County in Shandong Province at present,and to provide the scientific evidence for making strategies in prevention and control.Methods Children aged 8-12 years old and adults above 30 years old were selected from 8 endemic fluorosis villages in 2 fluorosis of children aged 8-12 years old were diagnosed by Dean method and skeletal fluomsis diagnosed by clinic and X-Rays.Results Eight villages in 2 towns were chosen underwent epidemiological investigation.Eight villages had water fluoride content＞4.50 mg/L.the highest water fluoride content was 5.78 mg/L.The total rate of dental fluorosis of children aged 8-12 years old WaS 90.70%(195/215),the index of dental fluorosis was 2.15 and the rate of dental damage was 24.65%(53/215).The rate of skeletal fluorosis detected by clinic and X-rays in adults older than 30 years old were 30.71%(78/254)and 16.54%(42/254),respectively.The averaged fuoride level in urine wa8 over 1.50 mg/L in 98.95%(189/191)of children aged 8-12 years old and in 97.92%(235/240)adults older than 30 years old,with the highest respectively being 14.50 mg/L and 17.99 mg/L.Conclusions In Boxing County in Shandong Province,endemic fluorosis is not effectively controlled.So endemic fluorosis control mfist be strengthened.

OBJECTIVERecently, a new traditional Chinese medicine differentiation theory "Syndrome Element (SE)" has been raised. In this study, the main syndrome element types and their correlations with the results of coronary angiography (CAG) in patients with coronary heart disease (CHD) were investigated.

METHODSEpidemiology cross-sectional study method was employed and 324 patients with CHD were enrolled, and their syndrome element types as well as the CAG results were analyzed. The correlations among syndrome element types, Gensini score, and the number of abnormal branches were also analyzed based on the distribution characteristics of syndrome element and coronary angiography results in the 324 cases.

RESULTSAccording to their occurrence frequency in 324 CHD patients, the top eight major heart syndrome elements were Xin () blood stasis (85.8%), Xin qi deficiency (79.6%), Xin heat blockage (41.1%), Xin phlegm with turbid fluid (38.0%), Xin qi stagnation (24.7%), Xin yang deficiency (18.9%), Xin yin deficiency (17.5%) and Xin cold coagulation (4.4%), respectively, which suggested that Xin blood stasis and Xin qi deficiency were the two most common syndrome elements. Also, as coronary artery Gensini score increased, the changing trend of the syndrome element was "Xin yang deficiency with blood stasis" to "Xin phlegm obstruction with heat blockage" to "Xin yin deficiency with blood stasis" to "Xin qi deficiency with blood stasis" to "Xin cold coagulation with phlegm and turbid fluid, "Xin cold coagulation with blood stasis" to "Xin deficiency of qi, yin and yang". As the number of abnormal branches increased, the syndrome element changing trend was "simultaneous occurrence of cold and heat syndrome" to "Xin qi and yang deficiency with blood stasis" to "Xin retention of phlegm with turbid fluid" to "Xin cold coagulation in the heart meridian", "Xin deficiency of both qi and yin". The result of this study shows that Xin qi deficiency and Xin blood stasis were the major syndrome elements in patients with CHD.

CONCLUSIONAs the severity and extent of coronary artery lesion increased, there were some apparent correlations among syndrome elements, Gensini score and number of abnormal coronary artery branches.

Coronary Angiography ; Coronary Disease ; diagnostic imaging ; physiopathology ; Cross-Sectional Studies ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Qi ; Yin-Yang