This study aimed at investigating the inhibitory effects and the anti-tumor mechanisms of co-adminis-tration of fusion proteins mGM-CSF/βhCG ( GC ) and hVEGF121/βhCG ( VC ) on RM-1 prostatic cancer and B16 F10 melanoma in the C57 BL/6 J mouse model. Two recombinant stains containing pET-28 a-mGM-CSF-X10-βhCGCTP37 and pET-28 a-VEGF-M2-X10-βhCG-CTP37 were induced by lactose to express fusion proteins. The fusion proteins were separated and purified to prepare the anti-tumor protein vaccines ( VC protein vaccine and GC protein vaccine) , which were then mixed to prepare a combined protein vaccine named VGC protein vac-cine. The prostatic cancer and melanoma tumor-bearing mice C57 BL/6 J were immunized with described vac-cines, then the growth of each tumor was measured;splenocyte proliferation of immunized mice was detected;and the cytotoxic effects of the vaccine on tumor cells were tested. After that, the in vivo concentrations of IFN-γ and anti-hVEGF antibodies were investigated by ELISA. The difference between each experimental group and normal saline group ( NS) was statistically significant in both tumor-bearing mouse models ( P <0. 05) respectively. Besides, VGC group exhibited significantly better anti-tumor effect compared with the GC and VC groups with the anti-tumor rate ( 41. 7 ± 0. 83)% and ( 46. 4 ± 1. 27)% for prostatic cancer and melanoma tumor, respectively. The co-administration of the two proteins, VC and GC, could inhibit the growth of RM-1 prostatic tumor and B16F10 melanoma effectively via anti-tumor immunity and anti-tumor angiogenesis.

Objective To investigate the incidence of nutritional risk for stroke patients with diabetes mellitus. Methods All the stroke patients with diabetes mellitus in a neurologic department were investigated with Nutritional Risk Screening (NRS2002). Results The incidences of undernutrition and nutritional risk were 11.3% (42/372) and 35.8% (133/372), respectively. The incidence of nutritional risk was more in the patients aged over 60 years than in the patients below 60 years (P=0.001). The incidence of both undernutrition and nutritional risk was more in the patients with stroke relapsing than those of the initial stroke (P<0.001). Conclusion The stroke patients with diabetes mellitus are in the risk of undernutrition and nutritional risk, especially those over the age of 60 years and relapsed stroke.

Autophagosomes derived from tumor cells have been proved to induce potent T cell response both in mouse and human. In human in vitro study, dendritic cells(DC)loaded with cytomegalovirus(CMV)pp65 antigen-containing DRibble were capable to efficiently re-stimulate pp65-specific T-cell recall responses from freshly isolated or frozen humanperipheral blood mononuclear cell(PBMC). This study developed more robust assays using in vitro expanded antigen-specific T cells that contained a much higher percentage of antigen-specific T cells. DC cross-presentation efficiency of OX40 and CD80 modified pp65-DRibble was detected by intracellular IFN-γ staining. Compared with Ctrl/pp65 DRibble, the percentage of IFN-γ+ in total CD8+ T cells andCD4+ T cells was improvedwith OX40/pp65 DRibbleand CD80/pp65 DRibble stimulation. In addition, vaccine induced IL-12indendritic cells, whichpolarizes Th cells toward the IFN-γ high Th1 phenotype, evaluated by ELISA inco-culture supernatantwas dramatically higher in OX40/pp65 DRibble and CD80/pp65 DRibblegroups than in Ctrl/pp65 DRibble group. These results have implications for the immuneactivity of OX40 and CD80 modified DRibble and choice for prospective clinical use ofDRibble-based cancer immunotherapy.

OBJECTIVETo assess the value of chromosome microarray analysis (CMA) for identifying the etiology of developmental delay/intellectual disability (DD/ID).

METHODSA total of 489 children referred for DD/ID with or without other abnormalities were recruited. All patients showed a normal karyotype. DNA was extracted and hybridized with Affymetrix CytoScan 750K array by following the manufacturer's protocol. The data was analyzed with CHAS v2.0 software.

RESULTSThe children were classified as with isolated DD/ID (n=358), DD/ID with epilepsy (n=49), and DD/ID with other structural anomalies (n=82). Pathogenic copy number variants (CNVs) were identified in 126 cases (25.8%), which included 89 (24.9%, 89/358) of whose with isolated DD/ID, 13 (26.5%, 13/49) of those with DD/ID and epilepsy, and 24 (29.3%, 24/82) of whose with DD/ID and other structural anomalies [P=0.064 (24.9% vs. 26.5%), P=0.679 (24.9% vs. 29.3%), and P=0.113 (26.5% vs. 29.3%), respectively]. Among the 126 cases, 79 were identified as microdeletion/microduplication syndromes, which included 15q24 microdeletion syndrome, Xq28 microduplication syndrome, and Lowe syndrome. Forty-seven cases had de novo pathogenic CNVs. ABAT, PMM2, FTSJ1, DYNC1H1 and SETBP1 were considered as candidate genes for DD/ID.

CONCLUSIONCMA is an effective method for identifying the etiology of DD/ID and is capable of identifying microdeletion/microduplication syndromes as well as de novo pathogenic CNVs which may be missed by conventional karyotyping. Based on the results, candidate genes for DD/ID may be identified.

Child ; Child, Preschool ; Chromosomes ; genetics ; Developmental Disabilities ; genetics ; Female ; Humans ; Infant ; Intellectual Disability ; genetics ; Karyotyping ; methods ; Male

To investigate the effects of Mycobacterium tuberculosis heat shock protein 65(HSP65)on Treg/Th17 immune balance in ApoE-knockout(ApoE-/-)mice, ApoE-/- mice with a high-cholesterol diet were immunized with M. tuberculosis HSP65. Sera were obtained for measurement of anti-HSP65 antibodies by ELISA; the effect of administration of different antigens was investigated, respectively, using flow cytometry analysis on the number of CD4+CD25+Foxp3+Tregs and CD4+IL-17+ Th17; the production of cytokines(IL-10, TGF-β1, IL-17 and IL-21)by these cells were determined by ELISA; total plasma cholesterol(TC), triglyceride(TG), high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)levels were detected by biochemical autoanalyzer. Atherosclerotic lesions were measured by lipid deposition stained with oil red O. The results demonstrated that the levels of anti-HSP65 IgG antibodies were increased significantly in Mycobacterium tuberculosis HSP65-treated ApoE-/- mice, revealed obvious decrease in Treg number, Treg related cytokines(IL-10, TGF-β1)levels and significant increase in Th17 number, Th17 related cytokines(IL-17 and IL-21)levels, the levels of TC, TG, HDL-C and LDL-C did not change between groups, while the atherosclerotic lesions significantly increased. Results indicate that M. tuberculosis HSP65 could interrupt the Th17/Treg immune balance in ApoE-/- mice, suggesting a potential role in the formation and progression of atherosclerosis.

An expression vector pET-28a-mGM-CSF-X10-βhCGCTP37 plasmid containing the βhCG and mGM-CSF gene was designed and constructed. The fusion protein was induced by lactose and purified by ammonium sulfate precipitation and DEAE-cellulose anion exchange column. Then dendritic cells(DC)in C57BL/6J mice were extracted and sensitized by the fusion protein to obtain DC vaccine. The DC vaccine was inoculated to C57BL of / 6J mice with prostate cancer RM-1. The results indicated that the anti-tumor effects of DC group and DC combined with paclitaxel(DP)group were superior to that of paclitaxel(Pac)group(P< 0. 01), and the anti-tumor effect of DP group was better than that of DC group. Thus, the constructed DC vaccine can inhibit the growth of prostate cancer, and have synergistic anti-tumor when used with paclitaxel.

In recent years, the demand of biologics has increased rapidly. Cell culture process with perfusion mode has become more and more popular due to its high productivity, good quality and high efficiency. In this paper, the unique operation and the details of process optimization for perfusion culture mode are discussed by comparing with traditional batch culture process. Meanwhile, the progress and strategies in the development and optimization of perfusion culture process in recent years are summarized to provide reference for the future development of mammalian cell perfusion culture technology.
Animals ; Batch Cell Culture Techniques ; trends ; Bioreactors ; standards ; CHO Cells ; Cricetulus ; Mammals ; Perfusion

ObjectiveTo investigate the epidemiological traits and potential years of life lost associated with lung cancer mortality among inhabitants of Shanghai's Pudong New Area from 1995 to 2021， in order to serve as a reference for developing intervention approaches. MethodsThe death surveillance system was used to gather statistics on lung cancer deaths. Crude mortality rate （CMR）， standardized mortality rate （SMR）， potential years of life lost （PYLL）， average years of life lost （AYLL）， annual percent change （APC） of the lung cancer deaths were analyzed. The impact of age-structural and non-age-structural factors on changes in lung cancer mortality was quantified using difference decomposition. ResultsThe CMR and SMR of lung cancer among residents in Pudong New Area between 1995 and 2021 were 58.21/10⁵ and 26.75/10⁵， respectively. The CMR of lung cancer increased over the years （APC=1.91%， 95%CI=1.60%‒2.30%； Z=11.487， P<0.001）， and the SMR of lung cancer declined over the years （APC=-1.50%， 95%CI=-1.80%‒-1.20%； Z=-9.006， P<0.001）. Age structure factors and gender factors contributed to the increase of lung cancer mortality， while non-population age structure factors overall appeared to play a protective role which might be related to the improvements in factors such as tobacco control and environmental management. The PYLL of lung cancer was 160 296 person years， the PYLL rate was 2.24‰， and the AYLL was 3.86 years per person. ConclusionAge structure factors are a significant contributor to the disease burden and result in the increase in the crude lung cancer mortality rate of Pudong residents of shanghai. Comprehensive monitoring， preventive， and control methods should be implemented.