AIM:To explore the promoting action of chloroquine on the anti-proliferation effect of dexametha-sone on acute lymphoblastic leukemia cells .METHODS:CCK-8 assay was used to assess the viability of the dexametha-sone-resistant human acute lymphoblastic leukemia CEM-C1 cell line treated with the combination of chloroquine and dexa-methasone .Western blotting , quantitative real-time PCR and LysoTracker Red staining were utilized to examine the mecha-nism.RESULTS:Combination of chloroquine and dexamethasone significantly inhibited the proliferation of CEM -C1 cells compared with control group (P<0.01).The combination of chloroquine and dexamethasone increased the abundance of glucocorticoid receptor and inhibited lysosomal function , while lysosomal inhibitor bafilomycin A 1 also increased glucocorti-coid signaling .CONCLUSION:Dexamethasone combined with chloroquine triggers an anti-proliferation effect on CEM-C1 cells via a lysosome-mediated pathway .

Objective To understand the characteristics of pathogenic bacteria and drug resistance of ventilator-associated pneu-monia(VAP)in neonatal intensive care unit (NICU),and to explore the corresponding prevention and control measures,and to pro-vide the basis for the VAP antibiotic treatment.Methods A total of 80 children with respiratory failure and ventilator assisted breathing were selected from the NICU as objects in this study.The clinical data,pathogenic bacteria and drug resistance were ana-lyzed retrospectively.Results In 80 cases,the incidence of VAP was 43.75% (35/80),a total of 70 strains of pathogenic bacteria were isolated,gram negative bacilli accounted for the highest proportion,accounting for 81.43% (57/70).Pseudomonas aeruginosa, Klebsiella pneumoniae were the most common gram negative bacilli.Gram positive cocci accounted for 8.57% (13/70),which domi-nated by methicillin resistant Staphylococcus aureus,in addition to susceptible to vancomycin,but resistant to the other antibiotics. Conclusion Gram negative bacillus are the main bacteria in VAP cases,and which are multiple drug-resistant pathogens.

Objective:To study the relationship between genetic polymorphisms of GSTM1 GSTT1 and the susceptibility of laryngeal and hypopharyngeal carcinomas(LHC).Method:The GSTM1 an GSTT1 genotypes were determined by multiplex PCR analysis in 76 LHC patients and 76 population controls.The association be tween the genotypes and LHC risk was measured by odds ratios(ORs)and 95% confidence intervals(95%Cls).Resuit:The frequency of GSTM1 null genotype was 59.2% in the LHC patients and 42.1% in controls(OR=1.935,95%CI=1.069-3.510),the difference was significant(P＜0.01).The frequency of GSTT1 null genotype was 57.9% in the LHC patients and 51.3% in controls.The difference was not significant(P＞0.05).In smokers,the risk of the LHC increased in subjects of GSTM1 null genotype(OR=5.545,95%CI=2.158-13.528).Conclusion:GSTM1 polymorphisms are associated with susceptibility to the LHC.It has the synergistic effects with smoking in the development of the LHC.GSTT1 genotypes might have no association with risk of the LHC in urban Linyi.

OBJECTIVE: To study the relationship between genetic polymorphisms of GSTM1 GSTT1 and the susceptibility of laryngeal and hypopharyngeal carcinomas (LHC). METHOD: The GSTM1 and GSTT1 genotypes were determined by multiplex PCR analysis in 76 LHC patients and 76 population controls. The association between the genotypes and LHC risk was measured by odds ratios (ORs) and 95% confidence intervals (95% CIs). RESULT: The frequency of GSTM1 null genotype was 59.2% in the LHC patients and 42.1% in controls (OR=1.935, 95% CI=1.069-3.510), the difference was significant (P<0.01). The frequency of GSTT1 null genotype was 57.9% in the LHC patients and 51.3% in controls. The difference was not significant (P>0.05). In smokers, the risk of the LHC increased in subjects of GSTM1 null genotype (OR=5.545, 95% CI=2.158-13.528). CONCLUSION GSTM1 polymorphisms are associated with susceptibility to the LHC. It has the synergistic effects with smoking in the development of the LHC. GSTT1 genotypes might have no association with risk of the LHC in urban Linyi.
Carcinoma, Squamous Cell ; genetics ; Case-Control Studies ; Disease Susceptibility ; Female ; Gene Frequency ; Genotype ; Glutathione Transferase ; genetics ; Humans ; Hypopharyngeal Neoplasms ; genetics ; Laryngeal Neoplasms ; genetics ; Male ; Polymorphism, Genetic

Objective To evaluate the inductive effect of interferon-γ(IFN-γ) combined with tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) on programmed necrosis of the human immortalized keratinocyte cell line HaCaT,and to explore its mechanisms.Methods In vitro cultured HaCaT cells were divided into several groups:negative control group receiving no treatment,IFN-γ group treated with 50 μg/L IFN-γ,TRAIL group treated with 4 μg/L TRAIL,and cytokine combination group treated with 50 μg/L IFN-γ and 4 μg/L TRAIL or zVAD combination group pretreated with 40 μmo/L zVAD for 1 hour followed by the treatment with 50 μg/L IFN-γand 4 μg/L TRAIL.After 48-hour treatment,the morphology of HaCaT cells were observed under a light microscope,methyl-thiazolyl-tetrazolium assay was performed to evaluate the inhibitory effect of the treatment on the proliferation of HaCaT cells,and double staining flow cytometry to detect the necrosis of HaCaT cells.Meanwhile,real-time fluorescence-based quantitative PCR (qPCR) was conducted to determine the mRNA expression of receptor interaction protein kinase 3 (RIP3) and mixed lineage kinase domain-like protein (MLKL),Western blot analysis to determine the expression of RIP1,RIP3,MLKL proteins and their phosphorylated forms (pRIP1,pRIP3,pMLKL),immunofluorescent staining to observe the distribution of pRIP3 and pMLKL in HaCaT cells,and the level of reactive oxygen species (ROS) in HaCaT cells in the above groups was detected by the fluorescence probe DCFH-DA.Statistical analysis was carried out with SPSS 22 software by using one-way analysis of variance (ANOVA) for comparing indices among different groups,and least significant difference (LSD)-t test for multiple comparisons.Results After 48-hour treatment,HaCaT cells in the cytokine combination group and zVAD combination group showed necrosis-like morphologic features.Methyl-thiazolyl-tetrazoliumassay revealed significant differences in the survival rate of HaCaT cells among the IFN-γgroup,TRAIL group,cytokine combination group,zVAD combination group and negative control group (73.16％ ± 5.71％,81.46％ ± 4.68％,72.18％ ± 2.93％,69.67％ ± 3.24％ and 100％,respectively;F =24.34,P ＜ 0.001).The necrosis rate of HaCaT cells was notably higher in the cytokine combination group and zVAD combination group (9.86％ ± 1.31％,10.33％ ± 2.16％,respectively) than in the negative control group (5.26％ ± 0.91％,t =4.61,5.07,respectively,both P ＜ 0.05).qPCR revealed that the mRNA expression of RIP3 and MLKL significantly increased in the cytokine combination group and zVAD combination group compared with the negative control group (tRIP3 =0.99,1.84,tMLKL =1.51,2.17,respectively,all P ＜ 0.05).Western blot analysis suggested that the protein expression of RIP1,RIP3,MLKL,pRIP1,pRIP3 and pMLKL significantly increased in the cytokine combination group compared with the negative control group (all P ＜ 0.05),and the zVAD combination group showed significantly decreased caspase 8 expression and increased expression of the above proteins compared with the cytokine combination group.Fluorescence microscopy showed that enhanced green dot-like or clump-like fluorescent spots (representing pRIP3) could be observed in the cytoplasm,and red fluorescent spots (representing pMLKL) could be seen on the cell membrane in the cytokine combination group.The average fluorescence intensity of ROS was significantly higher in the cytokine combination group than in the negative control group (t =702.00,P ＜ 0.05).Conclusion IFN-γcombined with TRAIL can induce the programmed necrosis of HaCaT cells with increased level of ROS.

Objective To investigate the correlation between the appendicular skeletal mass index(ASMI)and os-teoporosis(OP)in postmenopausal patients with type 2 diabetes(T2DM).Methods 164 hospitalized postmeno-pausal elderly T2DM patients were selected and their bone density(BMD)and appendicular skeletal mass were measured using dual energy X-ray absorption method(DXA).ASMI=appendicular skeletal mass/height2(kg/m2),and they were divided into OP group and non OP group based on T value.The general clinical data,blood biochemical indicators,and ASMI between the two groups were compared,and Logistic regression analysis,ROC curve were further used to analyze the correlation and diagnostic power.Results Compared with non OP group,OP group had a higher incidence of sarcopenia(SAC)(P＜0.05);the differences in age,ASMI,body mass index(BMI),and estradiol(E2)had significant differences between the two groups(P＜0.05);Logistic regression analysis showed that the reduction of ASMI[OR=0.133,95％CI(0.029-0.611)],BMI[OR=0.785,95％CI(0.625-0.985)],and E2[OR=0.967,95％CI(0.942-0.993)]were protective factors for OP;receiver operating curve(ROC)suggested that the AUC of ASMI for OP prediction was 0.752[95％CI(0.632-0.872),P＜0.001]with sensitivity 87.5％,specificity 47.8％,and the best diagnostic value was 5.52 kg/m2.Conclusion The reduction of ASMI,BMI,and E2 is positively correlated with the occurrence of osteoporosis.ASMI is an important protective factor for osteoporosis.Early OP screening and risk factor assessment should be conducted for elderly postmenopausal T2DM patients,and early intervention measures should be taken to reduce the risk of falls and fractures.

Autism spectrum disorder (ASD) is a highly heritable neurodevelopmental disorder characterized by deficits in social interactions and repetitive behaviors. Although hundreds of ASD risk genes, implicated in synaptic formation and transcriptional regulation, have been identified through human genetic studies, the East Asian ASD cohorts are still under-represented in genome-wide genetic studies. Here, we applied whole-exome sequencing to 369 ASD trios including probands and unaffected parents of Chinese origin. Using a joint-calling analytical pipeline based on GATK toolkits, we identified numerous de novo mutations including 55 high-impact variants and 165 moderate-impact variants, as well as de novo copy number variations containing known ASD-related genes. Importantly, combined with single-cell sequencing data from the developing human brain, we found that the expression of genes with de novo mutations was specifically enriched in the pre-, post-central gyrus (PRC, PC) and banks of the superior temporal (BST) regions in the human brain. By further analyzing the brain imaging data with ASD and healthy controls, we found that the gray volume of the right BST in ASD patients was significantly decreased compared to healthy controls, suggesting the potential structural deficits associated with ASD. Finally, we found a decrease in the seed-based functional connectivity between BST/PC/PRC and sensory areas, the insula, as well as the frontal lobes in ASD patients. This work indicated that combinatorial analysis with genome-wide screening, single-cell sequencing, and brain imaging data reveal the brain regions contributing to the etiology of ASD.
Humans ; Autism Spectrum Disorder/metabolism* ; Autistic Disorder ; Exome Sequencing ; DNA Copy Number Variations ; East Asian People ; Brain/metabolism* ; Mutation/genetics* ; Genetic Predisposition to Disease/genetics*