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 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 channeland acetaldehyde in arrhythmia caused by acute alcoholic intoxication.Methods Primary 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).Conclusions Acute 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.

Objective:To develop a virtual reality (VR) exercise rehabilitation system for home-based patients with spinal cord injury, and test patients' acceptance and experience.Methods:The VR exercise rehabilitation system for home-based patients with spinal cord injury was developed by a multidisciplinary team based on the home rehabilitation needs, evidence, and clinical experience of spinal cord injury patients, and further improved after recommendations from experts and patients. From February to July 2024, convenience sampling was used to select 148 patients with spinal cord injuries admitted to the Department of Spinal Cord and Neurological Function Reconstruction at the China Rehabilitation Research Center. Acceptance Questionnaire was used to investigate patients' acceptance of the system. Thirteen patients with spinal cord injuries were selected for semi-structured interviews to explore the themes of their experiences using VR exercise rehabilitation system for home-based patients with spinal cord injury.Results:A VR exercise rehabilitation system for home-based patients with spinal cord injury was developed, which includes four aspects of personal profile, personal assessment, training selection, and care knowledge. The total acceptance score of this system among 148 patients was (87.69±1.59). Through semi-structured interviews, three themes were identified involving excellent experience, expectation of rehabilitation effects, satisfaction with functionality and hope for continuous system updates.Conclusions:The design of VR exercise rehabilitation system for home-based patients with spinal cord injury is scientifically reasonable, which can further verify the rehabilitation effect of the system.

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:Healthy life expectancy (HLE), which combines life expectancy with health, is an essential comprehensive measure of life length and quality. This article aimed to systematically review the methods for defining and measuring HLE and describe application studies published, providing a reference for decision makers to select and develop methods suitable for China's conditions to measure HLE.Methods:Seven Chinese and English literature databases were searched up to May 7, 2022, and several related reviews and bibliography were manually retrieved. Systematic reviews and empirical research were included concerning HLE indicators and measurement of HLE. Information including the study area, type of the study, study population, HLE index, measurement method, data sources, and results from application studies published in the last five years were extracted. The evolution of the definition of HLE, the scope of different indicators, the measurement scale of health, and measurement methods, were all collected. Results of the empirical research related to measurement methods of indicators were summarized. The study followed the scoping review framework and was written according to the PRISMA-ScR statement.Results:A total of 84 articles were included, including 13 reviews, 17 original studies related to HLE index definition, ten original studies related to index measurement, and 44 empirical studies conducted in the past five years. There were as many as 20 indicators related to HLE, and each scale had its emphasis. A total of ten methods measuring HLE were identified, which vary in the definition of health, whether using weight, and the data type. The most commonly used indicators in the past five years were disability-free life expectancy and HLE. For the method of HLE calculation, Sullivan's method was mainly used for cross-sectional data, and the multistate life table was mainly used for longitudinal data.Conclusions:There are various definitions and measurement methods of HLE, but none are suitable for all scenarios. To summarize the HLE concept, health evaluation techniques, measurement methods, and application studies published worldwide can provide a reference for the localization of HLE measurement in China.

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*