Objective To study the technology of loading trehalose into cytoplasm of human platelets before lyophilization,and to determine the optimal experimental conditions of pretreatment technology.Methods The curves of loading efficiency,the internal trehalose concentration versus incubated time,temperature,and external trehalose concentration were drawn up to screen the optimal loading parameters.Results The loading efficiency was linear to the incubated time(2 hours later) and incubated temperature(range from 30℃ to 40℃),respectively.The loading efficiency almost reached 60 percent when the platelets were incubated at 37℃ for 4h.The intracellular trehalose concentration was increased with the raise of the external trehalose concentration(

Objective: To study the diagnostic characteristics of cardiac amyloidosis (CA) by non-invasive electrocardiography (ECG) in relevant patients. Methods: We retrospectively analyzed 60 CA patients diagnosed in our hospital from 2008-08 to 2013-12 for their clinical and ECG characteristics. Results: There were 48 male and 12 female patients with the ratio of 4: 1. The ifrst time diagnosis rate was low and the average age for conifrmed diagnosis was at (54. 5±14. 2) years.①There were 32 (53. 3%) cases combining heart failure, 12 (20%) with pleural effusion, 20 (33. 3%) with atrial arrhythmia, 8 (13. 3%)with ventricular arrhythmia, 4 (6. 7%)with sino-atrial block, 15 (25%)with atrio-ventricular block, 4 (6. 7%) with left bundle branch block (LBBB), 5 (8. 3%)with RBBB and 8 (13. 3%)with intra-ventricular block.②There were 32 (53. 3%) cases with low voltage on limb leads, 52 (86. 7%) with pseudo-infarct pattern, 48 (60%) with ST-T abnormality and 30 (50%) combining low voltage on limb leads with pseudo-infarct pattern.③The patients combining pleural effusion and with pseudo-infarct pattern had the increased ratio of low voltage on limb leads, while there were still 22 (45. 8%) cases without pleural effusion had low voltage on limb leads.④ ECG characteristics for 60 CA patients were as follows: QRS duration (104±26) ms, QT interval (404±34) ms, QTc (462±35) ms; the R wave of avR 0. 17 mV, QRS wave 0.30 mV; the R wave of limb leads and V1-3 were all<0.5mV, the S wave of V1-3 were 0. 62mV, 1. 61mV, 1. 56mV; the R/S ratio of V1-3 were 0. 19, 0. 12, 0. 20 respectively. Conclusion: CA patients had the highest incidence of pseudo-infarct pattern; meanwhile, combining with low voltage on limb leads, pseudo-infarct with long Q or S wave and ST-T abnormality but normal QRS duration was helpful for differential diagnosis of CA in clinical practice.

Gastrointestinal cancers are the common malignant tumors of the digestive system, and their morbidity and mortality are in the forefront of malignant tumors. Currently, cancer immunotherapy is the hottest topic in cancer research field. Although cancer immunotherapy has achieved some results in the fundamental research and clinical application of gastrointestinal tumors, there are still a series of problems that need to be resolved. In this article, we review the fundamental and clinical research progress of several common methods of cancer immunotherapy in the field of gastrointestinal tumors.

This study aimed at evaluating the antioxidant effects of Guanfu base A(GFA)on acetylcholine(Ach)/CaCl2(CaCl2 10 mg/mL, Ach 66 μg/mL)-induced atrial fibrillation(AF)in rats. SD rats were rando-mized into normal group, model group, GFA treatment groups(6 mg/kg, 12 mg/kg), Amiodarone(Ami)treatment group(50 mg/kg)and Lovastatin(Lov)treatment group(10 mg/kg). The AF durations were measured by electrocardiogram(ECG). The effective refractory periods(AERP)were measured in the left atrial appendage. Oxidative stress-related gene and protein expression was evaluated by RT-PCR and Western blot. The activity of antioxidant enzymes was measured by enzymatic assay. Results indicated that, in comparison with that in the vehicle-treated AF rats, treatment with GFA(6 mg/kg, 12 mg/kg, po), significantly shortened the AF duration and prolonged the AERP in rats. In addition, treatment with GFA reduced the levels of plasma and myocardium malondialdehyde, increased the activity of plasma superoxide dismutase in a dose-dependent manner. Moreover, treatment with GFA mitigated AF-up-regulated p22phox, p47phox, gp91phox, and p67phox NADPH oxidase expression, and AF-increased ratios of membrane to cytosolic Rac-1 in the atrium. It also significantly prevented AF-down-regulated atrial connexin40 expression in rats. Data suggested that GFA(6 mg/kg, 12 mg/kg)has potent anti-oxidant activity and inhibits oxidative-stress-related AF in rats.

Objective:To detect and analyze the gene variation types of 64 unrelated pedigrees affected with autosomal dominant polycystic kidney disease (ADPKD), and explore the detection efficiency of multiple gene analysis techniques and variation characteristics.Methods:It was a cross-sectional study. The clinical data of 64 pedigrees with ADPKD from Nephrology Department or Genetic and Prenatal Diagnosis Center of the First Affiliated Hospital of Zhengzhou University from December 2017 to August 2020 were retrospectively analyzed. The blood samples of probands and other family members were collected. Genetic analysis was carried out by next generation sequencing, and suspected mutations were verified by multiplex ligation-dependent probe amplification, or long-range PCR combined with Sanger sequencing. Prenatal diagnosis for high-risk fetuses was performed by fetal villi or amniotic fluid cells after genotyping without maternal genomic DNA contamination.Results:Among detected 64 pedigrees, 57 pedigrees (89.06%) had genetic variants in PKD1/PKD2. A total of 49 pathogenic/likely pathogenic variants in PKD1/PKD2 were identified in 51 pedigrees (79.69%), including 14 nonsense variants (28.57%), 14 frameshift variants (28.57%), 11 missense variants (22.45%), 5 splicing variants (10.20%) and 5 deletion variants (10.20%). Of these variants, 87.76% (43/49) were in PKD1 and 12.24% (6/49) were in PKD2. Totally, 14 novel variants in PKD1/ PKD2 were identified, including 7 frameshift variants, 3 splicing variants, 2 nonsense variants, 1 deletion variant and 1 missense variant, of which 11 variants were in PKD1 and 3 variants were in PKD2. Twenty high-risk fetuses from 17 pedigrees received prenatal diagnosis, in whom 6 fetuses had PKD1 variation, and other 14 fetuses had no PKD1/ PKD2-genetic variation. Conclusions:The combination of next-generation sequencing, multiplex ligation-dependent probe amplification, and long-range PCR combined with Sanger sequencing can be helpful for rapid, efficient and accurate genetic diagnosis of ADPKD pedigrees. Point mutations are the most common types in PKD1/PKD2. Fourteen novel variants in PKD1/PKD2 extend its pathogenic variant spectrum and can provide basis for genetic counseling and prenatal diagnosis of ADPKD pedigrees.

Objective:To explore the interaction effects on brain activity between the smoking addiction and weight status by resting-state functional magnetic resonance imaging.Methods:Retrospective analysis of clinical data and resting-state functional magnetic resonance imaging data were analyzed from 99 recruited subjects from January 2019 to December 2021. All participants were divided into four groups: overweight smokers ( n=24), normal-weight smokers ( n=28), overweight non-smokers ( n=19), and normal weight non-smokers ( n=28). Calculate regional homogeneity (ReHo) to reflect the internal brain activity of the subjects. Two-way ANOVA was used to detect the interaction effects between smoking addiction and overweight on ReHo by SPM12 software, correcting for age, years of education and head motion. Results:The interaction effect between smoking addiction and overweight on ReHo was significant in right superior frontal gyrus(x, y, z=15, 9, 60)(GRF corrected, Pvoxel<0.005, Pcluster<0.05). The ReHo value in the right superior frontal gyrus of overweight smokers was significantly higher than that of normal weight smokers ( t=3.768, P<0.001, Bonferroni corrected). The ReHo values in the right superior frontal gyrus of overweight non-smokers were significantly lower than those of normal weight non-smokers ( t=-3.242, P=0.002, Bonferroni corrected). The ReHo values in the right superior frontal gyurs of normal-weight smokers were significantly lower than those of normal weight non-smokers( t=-3.540, P=0.001, Bonferroni corrected). The ReHo values in the right superior frontal gyrus of overweight smokers were significantly higher than those of overweight non-smokers ( t=3.392, P=0.002, Bonferroni corrected). Correlation analyses showed that the strengthen ReHo value in right superior frontal gyrus was positively associated with pack-year in smoking addicts( r=0.387, P=0.007, Bonferroni corrected). Conclusion:Smoking addiction and overweight have an antagonistic effect on brain activity in the right superior frontal gyrus, which may provide potential therapeutic targets for individuals with comorbidity of smoking addiction and overweight.

OBJECTIVE: To explore the clinical and genetic features of a child with autosomal dominant mental retardation type 40 (MRD40) due to variant of the CHAMP1 gene. METHODS: Clinical characteristics of the child were analyzed. Genetic testing was carried out by low-depth high-throughput and whole genome copy number variant sequencing (CNV-seq) and whole exome sequencing (WES). A literature review was also carried out for the clinical phenotype and genetic characteristics of patients with MRD40 due to CHAMP1 gene variants. RESULTS: The child, a 11-month-old girl, has presented with intellectual and motor developmental delay. CNV-seq revealed no definite pathogenic variants. WES has detected the presence of a heterozygous c.1908C>G (p.Y636*) variant in the CHAMP1 gene, which was carried by neither parent and predicted to be pathogenic. Literature review has identified 33 additional children from 12 previous reports. All children had presented with developmental delay and mental retardation, and most had dystonia (94.1%), delayed speech and/or walking (85.2%, 82.4%) and ocular abnormalities (79.4%). In total 26 variants of the CHAMP1 gene were detected, with all nonsense variants being of loss-of-function type, located in exon 3, and de novo in origin. CONCLUSION The heterozygous c.1908C>G (p.Y636*) variant of the CHAMP1 gene probably underlay the WRD40 in this child. Genetic testing should be considered for children featuring global developmental delay, mental retardation, hypertonia and facial dysmorphism.
Humans ; Intellectual Disability/genetics* ; Genetic Testing ; Phenotype ; Exome Sequencing ; Heterozygote ; Mutation ; Chromosomal Proteins, Non-Histone/genetics* ; Phosphoproteins/genetics*

OBJECTIVETo identify the effect of ethanol and its metabolite acetaldehyde on acetylcholine-sensitive K(+) channel Kir3.1 protein expression, and explore the potential role of this channel and acetaldehyde in arrhythmia caused by acute alcoholic intoxication.

METHODSPrimary atrial cardiomyocytes were isolated from 150 newborn SD rats by typsin and type II collagenase, cultured and troponin I was determined by immunofluorescence. Cell survival in 200-800 mmol/L ethanol or 50-500 µmol/L acetaldehyde treated cells for 24 hours was measured by CCK-8 assay to determine the concentration of ethanol and acetaldehyde for inducing apoptosis in cardiomyocytes. The highest non-apoptotic concentration (200 mmol/L) of ethanol and acetaldehyde (100 µmol/L) was used in the main study. Kir3.1 protein expression was detected by Western blot.

RESULTS(1) Cellular immunofluorescence results showed that cultured cells are cardiomyocytes, and more than 90% of these cells are troponin I positive. (2) CCK-8 assay demonstrated that the survival rate of cardiomyocytes in the groups treated by ethanol over 400 mmol/L for 24 hours or acetaldehyde over 400 µmol/L was significantly lower than that of the control group (P < 0.05), while the survival rate was similar in cardiomyocytes treated by ethanol less than 200 mmol/L or acetaldehyde less than 350 µmol/L for 24 hours and the control group (P > 0.05). (3) Western-bolt assay revealed that ethanol and acetaldehyde treatment for 24 hours upregulated Kir3.1 protein expression in primary atrial cardiomyocytes of newborn SD rats by (44.52 ± 23.07)% and (45.04 ± 22.01)% respectively compared with the control group (all P < 0.01).

CONCLUSIONSAcute ethanol and acetaldehyde treatment could significantly upregulate the protein expression of acetylcholine-sensitive K(+) channel Kir3.1, this might serve as a potential mechanism for arrhythmia caused by acute alcoholic intoxication.

Acetaldehyde ; metabolism ; Acetylcholine ; Alcoholic Intoxication ; metabolism ; Animals ; Apoptosis ; Cells, Cultured ; Ethanol ; pharmacology ; G Protein-Coupled Inwardly-Rectifying Potassium Channels ; drug effects ; Heart Atria ; Myocytes, Cardiac ; drug effects ; Rats ; Rats, Sprague-Dawley ; Sincalide

Background: The incidence of ulcerative colitis (UC) has gradually increased in China in recent years. The pathogenesis of UC is related to the dysfunction of immune system. B7-H5 is an important immune checkpoint molecule and is significant for the regulation of immune function. Ainis: To investigate the expression and clinical significance of B7-H5 in UC. Methods: A total of 65 UC tissue specimens were collected from Jan. 2010 to Dec. 2020 at the First Affiliated Hospital of Soochow University, and 5 healthy subjects were served as controls. Immunohistoehemistry and immunofluorescence were used to detect the expression of B7-H5, and its relationship with elinieopathologieal characteristics of UC patients was analyzed. Results: Expression of B7-H5 was significantly increased in UC patients than in controls (P < 0. 001). B7-H5 expression in UC patients was positively correlated with ESR and CRP (P < 0. 01), but not related to gender, age, extent of lesion, Mayo score and UCEIS score (P > 0. 05). Conclusions; The expression of B7-H5 in UC patients is significantly increased and is correlated with ESR and CRP, and can be used as a new marker for reflecting the severity of inflammation in UC patients.

Acyl-CoA synthetase long chain family member 5 (ACSL5), is a member of the acyl-CoA synthetases (ACSs) family that activates long chain fatty acids by catalyzing the synthesis of fatty acyl-CoAs. The dysregulation of ACSL5 has been reported in some cancers, such as glioma and colon cancers. However, little is known about the role of ACSL5 in acute myeloid leukemia (AML). We found that the expression of ACSL5 was higher in bone marrow cells from AML patients compared with that from healthy donors. ACSL5 level could serve as an independent prognostic predictor of the overall survival of AML patients. In AML cells, the ACSL5 knockdown inhibited cell growth both in vitro and in vivo. Mechanistically, the knockdown of ACSL5 suppressed the activation of the Wnt/β-catenin pathway by suppressing the palmitoylation modification of Wnt3a. Additionally, triacsin c, a pan-ACS family inhibitor, inhibited cell growth and robustly induced cell apoptosis when combined with ABT-199, the FDA approved BCL-2 inhibitor for AML therapy. Our results indicate that ACSL5 is a potential prognosis marker for AML and a promising pharmacological target for the treatment of molecularly stratified AML.
Humans ; Antineoplastic Agents/therapeutic use* ; Apoptosis ; beta Catenin/metabolism* ; Biomarkers, Tumor/metabolism* ; Cell Line, Tumor ; Coenzyme A Ligases/metabolism* ; Leukemia, Myeloid, Acute/metabolism* ; Lipoylation ; Prognosis ; Wnt Signaling Pathway