Objective To investigate the infectivity of hepatitis A virus(HAV)cell-adapted strain in a type Ⅰ interferon receptor-deficient mouse model.Methods The biological charateristics of HM175/18f were identified,including the viral protein expression and viral proliferation by indirect immunofluorescence,Western blot and real-time quantitative RT-PCR in vitro.Then,type Ⅰ interferon receptor-deficient A129 mice were infected with HM175/18f via intravenous injection.The viral RNA load in serum,feces and liver tissues of infected mice were detected to determine the replication of HAV in vivo.The level of serum alanine aminotransferase(ALT)and HE staining of liver tissues were used to evaluate liver injury.Additionally,the dynamic changes of HAV-specific IgG antibody was detected to assess the humoral immune response induced by HM175/18f.Results A129 mice infected with HM175/18f did not show obvious clinical symptoms,nor was the ALT level significantly elevated.However,viral RNA persisted in the liver tissue of infected mice until 42 days after infection.There was focal infiltration of lymphocytes and neutrophils in the liver tissue of infected mice,but no focal necrosis was observed.More importantly,HM175/18f infection caused significant viremia and sustained fecal virus shedding.In addition,HM175/18f induced a significant HAV-specific humoral immune response in A129 mice.Conclusion Our study has revealed the infectivity of HAV cell-adapted strain HM175/18f in type Ⅰ interferon receptor-deficient mice,and identified the attenuated characteristics of HM175/18f,which not only contributes to our understanding of the pathogenesis of HAV,but also expand the applications of a type Ⅰ interferon receptor-deficient mouse model in the study of hepatitis A.

Objective:To investigate the incidence rate and gene variation of methylmalonic academia (MMA) in Ji′nan city by analyzing biochemical and genetic screening results, and to explore the carrier frequency of MMA-related pathogenic genes in the population in Ji′nan.Methods:The children diagnosed with MMA by tandem mass spectrometry screening in Ji′nan Neonatal Disease Screening Centre from May 2011 to May 2022 were enrolled in this study.Their genetic test results were retrospectively analyzed and summarized.The dried heel blood tablets collected from 6 800 newborns were tested for neonatal gene screening. MMAA, MMAB, MMACHC and MMUT genes in 4 800 cases were detected by high-throughput sequencing+ target area capture technology.Ultra-multiplex polymerase chain reaction+ target gene locus capture technology was used to detect 174 target loci of 8 genes related to MMA in 2 000 cases.The hotspot mutation and related gene carrier rate of MMA were analyzed. Results:A total of 367 452 newborns were screened by tandem mass spectrometry, and 103 cases (56 males and 47 females) were diagnosed with MMA by screening.The estimated incidence of MMA was 1∶3 567.Among the 103 MMA cases, 76 were genetically diagnosed, and 4 gene variants of MMA ( MMAHC, MMUT, MMAA, MMADHC) were identified.A total of 6 800 neonates underwent neonatal genetic screening.Three of them were diagnosed with MMA.About 318 infants carried pathogenic variants of MMA, with a total carrier rate of 4.68%.Specifically, the carrier rates of MMACHC and MMUT gene variants were 3.09%(210/6 800) and 1.43% (97/6 800), respectively. Conclusions:MMA is the most common organic acid metabolism disorder in our country.The incidence and carrier rate of this disease are high in Jinan city.Neonatal genetic screening is an important supplement to neonatal biochemical screening.Carrier screening for MMA-related pathogenic genes is recommended for couples of childbearing age in Jinan.

Aging/metabolism* ; Animals ; Gene Expression Regulation ; Macaca fascicularis ; Retina/metabolism* ; Single-Cell Analysis

Objective:To describe the prevalence of heart failure in China and to explore the prospective association between smoking behavior and the risk of incident heart failure.Methods:The subjects were from the China Kadoorie Biobank (CKB) and the baseline survey was conducted from June 2004 to July 2008. A total of 487 197 subjects were included in this study, after excluding those with missing BMI information, lost follow-up immediately after baseline investigation, and self-reported coronary heart disease, stroke, or malignant tumor at baseline. This study included data from baseline and follow-up until December 31, 2016. Cox proportional hazards regression models were used to estimate the association between smoking behavior and the risk of heart failure.Results:The median follow-up time was 10.15 years, during which a total of 4 208 new cases of heart failure occurred, with a crude incidence rate of 0.87/1 000 person-years and a cumulative incidence rate of 0.86%. The higher the age at baseline, the higher the incidence of heart failure. The incidence of heart failure in high age group, rural area and male was higher than that in low age group, urban area and female population respectively. Compared with non-smokers, there was no significant difference in the risk of heart failure in occasional smokers ( HR=1.05; 95% CI: 0.91-1.22), while former smokers ( HR=1.48; 95% CI:1.31-1.67) and current smokers ( HR=1.34;95% CI:1.22-1.49) increased risk. Former smokers ( HR=1.33;95% CI:1.21-1.46) and current smokers ( HR=1.46; 95% CI:1.31-1.64) had higher risk of heart failure than non-smokers or occasional smokers. No dose-response relationship was observed between the number of cigarettes smoked per day and the risk of heart failure in current and former smokers (for trend P=0.347 and 0.066). Compared with non-smokers or occasional smokers, the hazard ratios of <5, 5-, 10- and ≥20 years since quit smoking were 1.61 (95% CI: 1.36-1.92), 1.55 (95% CI: 1.27-1.90), 1.24 (95% CI: 1.02-1.51) and 1.35 (95% CI: 1.08-1.68), respectively (for trend P=0.091). The hazard ratios of quitting smoking due to disease and other reasons were 1.62 (95% CI:1.41-1.86) and 1.23 (95% CI: 1.04-1.45). Healthy smoking behaviors had a significant protective effect on heart failure compared with non-healthy smoking behaviors ( HR=0.75, 95% CI:0.69-0.81). Area and family history of coronary heart disease, and the smoking behaviors interacted with the risk of heart failure (for all interactions were P<0.05). Conclusions:The incidence of heart failure in China is higher in males than females, higher in rural areas than in urban areas, and increases with age. Both former smokers and current smokers had a higher risk of heart failure than nonsmokers or occasional smokers, regardless of the frequency, amount, duration, and reason for quitting. Smoking is an important risk factor for heart failure and comprehensive anti-smoking measures should be maintained.

Along with the significant development on both theory and practice of health promotion programs, the application of behavioral and social science theories has also been advanced in the fields of design and evaluation regarding the intervention-related studies. Intervention mapping is a new planning protocol, efficiently used to develop, implement, and evaluate health promotion related intervention programs. In this article, we are briefly introducing the basic concepts, implementation steps, specific requirements, as well as reviewing the current progress in methodologies, application that are related to intervention mapping, so as to provide reference for health intervention research studies, domestically.

Aging increases the risk of various diseases. The main goal of aging research is to find therapies that attenuate aging and alleviate aging-related diseases. In this study, we screened a natural product library for geroprotective compounds using Werner syndrome (WS) human mesenchymal stem cells (hMSCs), a premature aging model that we recently established. Ten candidate compounds were identified and quercetin was investigated in detail due to its leading effects. Mechanistic studies revealed that quercetin alleviated senescence via the enhancement of cell proliferation and restoration of heterochromatin architecture in WS hMSCs. RNA-sequencing analysis revealed the transcriptional commonalities and differences in the geroprotective effects by quercetin and Vitamin C. Besides WS hMSCs, quercetin also attenuated cellular senescence in Hutchinson-Gilford progeria syndrome (HGPS) and physiological-aging hMSCs. Taken together, our study identifies quercetin as a geroprotective agent against accelerated and natural aging in hMSCs, providing a potential therapeutic intervention for treating age-associated disorders.

Parthenogenetic embryonic stem cells (pESCs) derived from bi-maternal genomes do not have competency of tetraploid complementation, due to lacking of paternal imprinting genes. To make pESCs possess fully development potentials and similar pluripotency to zygote-derived ESCs, we knocked out one allelic gene of the two essential maternal imprinting genes (H19 and IG) in their differentially methylated regions (DMR) via CRISPR/Cas9 system and obtained double knock out (DKO) pESCs. Maternal pESCs had similar morphology, expression levels of pluripotent makers and in vitro neural differentiation potentials to zygotes-derived ESCs. Besides that, DKO pESCs could contribute to full-term fetuses through tetraploid complementation, proving that they held fully development potentials. Derivation of DKO pESCs provided a type of major histocompatibility complex (MHC) matched pluripotent stem cells, which would benefit research in regenerative medicine.
Animals ; Embryonic Stem Cells ; Gene Knockout Techniques ; Genomic Imprinting ; Mice ; Parthenogenesis ; Pluripotent Stem Cells ; Regenerative Medicine ; Tetraploidy

Objective: To investigate the effects of AZD5363, an inhibitor of protein kinase B (Akt), on proliferation, migration and apoptosis of human breast cancer cell line MDA-MB-231, and to further clarify their possible molecular mechanisms Methods: After treatment with different concentrations (0.5, 1, 5, 10, 20 and 50 μmol/L) of AZD5363, the viability of MDA-MB-231 cells was detected by MTT assay, the cell cycle distribution was analyzed by FCM, the cell migration ability was detected by wound healing test and Transwell chamber assay, the cell apoptosis rate was detected by TUNEL method. Then the expression levels of cell cycle- and apoptosis-related proteins were measured by Western blotting. Results: AZD5363 suppressed the cell viability in a dose-dependent manner (P < 0.05), and arrested the cell cycle progression at S phase by up-regulating the expression of p53 and down-regulating the expression of cyclin B1 (all P < 0.05). AZD5363 significantly inhibited the cell migration (P < 0.05), and induced the cell apoptosis (P < 0.05) by activating caspase-3 and poly (ADP-ribose) polymerase (PARP) proteins (both P < 0.05). Conclusion: AZD5363 can inhibit cell activity and migration, and induce apoptosis of human breast cancer cell line MDA-MB-2 31, thereby exhibiting its anticancer activity.

AIM:To investigate the effect of perifosine, an inhibitor of protein kinase B ( PKB/Akt) , on the cell cycle, apoptosis and autophagy in human brain glioma U251 cells, and to determine the relationship between perifos-ine-induced autophagy and apoptosis of glioma.METHODS:The cell growth inhibition was determined by MTT assay. The cell cycle distribution of U251 cells was examined by flow cytometry.The cell apoptosis was analyzed by Annexin V-FITC apoptosis detection kit.The protein expression of P21, P27, cyclin B1, caspase-9 and PARP was examined by Wes-tern blot analysis.The distribution and expression of LC3-Ⅱ, an autophagy marker, was observed to determine the effect of perifosine-induced autophagy.RESULTS:Perifosine inhibited the cell viability in a dose-dependent manner.In perifos-ine-treated U251 cells, the cell cycle was arrested in G2 phase and the expression of cyclin B1 was inhibited.Perifosine in-duced apoptosis of U251 cells through activation of caspase-9 cleavage, PARP cleavage and survivin inhibition.In addi-tion, suppression of autophagy by chloroquin, an inhibitor of autophagy, increased the number of apoptotic cells.CON-CLUSION:Perifosine inhibits cell proliferation and triggers apoptosis and autophagy in human U251 cells.Blocking auto-phagy magnifies perifosine-induced glioma cell apoptosis.

Objective To evaluate radical gastrectomy combined with cholecystectomy for gastric cancer patients with concomitant gallbladder disease.Methods Clinical data of 702 gastric cancer patients undergoing radical gastrectomy (614 patients) only or combined with cholecystectomy during radical gastrectomy from October 2009 to September 2014 in our department was retrospectively analyzed.Results The operating time of patients with simultaneous cholecystectomy was(348 ± 111)min.the operating time of patients with radical gastrectomy only was (274 ± 89) min (t =3.812,P ＜ 0.05).Perioperative and postoperative complications,hospitalization expenses and 5-year survival rates were not statistically significant (P ＞ 0.05).Conclusions Radical gastrectomy with cholecystectomy for gastric cancer with gallbladder disease patients is safe and feasible.