OBJECTIVE: This study aimed to explore the association between body mass index (BMI) and mortality based on the 10-year population-based multicenter prospective study. METHODS: A general population-based multicenter prospective study was conducted at four sites in rural China between 2013 and 2023. Multivariate Cox proportional hazards models and restricted cubic spline analyses were used to assess the association between BMI and mortality. Stratified analyses were performed based on the individual characteristics of the participants. RESULTS: Overall, 19,107 participants with a sum of 163,095 person-years were included and 1,910 participants died. The underweight (< 18.5 kg/m ²) presented an increase in all-cause mortality (adjusted hazards ratio [ aHR] = 2.00, 95% confidence interval [ CI]: 1.66-2.41), while overweight (≥ 24.0 to < 28.0 kg/m ²) and obesity (≥ 28.0 kg/m ²) presented a decrease with an aHR of 0.61 (95% CI: 0.52-0.73) and 0.51 (95% CI: 0.37-0.70), respectively. Overweight ( aHR = 0.76, 95% CI: 0.67-0.86) and mild obesity ( aHR = 0.72, 95% CI: 0.59-0.87) had a positive impact on mortality in people older than 60 years. All-cause mortality decreased rapidly until reaching a BMI of 25.7 kg/m ² ( aHR = 0.95, 95% CI: 0.92-0.98) and increased slightly above that value, indicating a U-shaped association. The beneficial impact of being overweight on mortality was robust in most subgroups and sensitivity analyses. CONCLUSION This study provides additional evidence that overweight and mild obesity may be inversely related to the risk of death in individuals older than 60 years. Therefore, it is essential to consider age differences when formulating health and weight management strategies.
Humans ; Body Mass Index ; China/epidemiology* ; Male ; Female ; Middle Aged ; Prospective Studies ; Rural Population/statistics & numerical data* ; Aged ; Follow-Up Studies ; Adult ; Mortality ; Cause of Death ; Obesity/mortality* ; Overweight/mortality*

Objective To establish a simple technical procedure for preparing doggybone DNA(dbDNA),and to discuss the distinctions in gene expression and immune response between dbDNA and conventional plasmid DNA.Induced pluripotent stem cell(iPSC)was generated from human peripheral blood mononuclear cells(PBMCs)by using dbDNA.To investigate a protocol for inserting ultra-long DNA fragments into the iPSC adeno-associated virus site 1(AAVS1),concurrently with the integration of the Cas9 expression frame point into the dbDNA-derived iPSC AAVS1 safe harbor,so as to establish a stable cell line that can achieve iPSC gene editing exclusively through the introduction of the single guide RNA(sgRNA)for the targeted gene.Methods A simple dbDNA preparation technique was established through utilization of Phi29 DNA polymerase multiple displacement amplification,protelomerase TelN digestion and covalent ligation,as well as the subsequent removal of the back-bone loop through the action of a restriction enzyme and exonuclease.The efficiency of green fluorescent protein(GFP)was eval-uated following electroporation of the dbDNA-GFP vector and the pMax-dbDNA-GFP traditional plasmid vector into PB-MCs.The mRNA expression of interferon-γ(IFN-γ)in the two groups was quantified by qPCR.The dbDNA reprogramming vectors dbDNA-MOS,dbDNA-KLF4,dbDNA-MYC and dbDNA-BCL were prepared and subsequently transferred into PBMCs to induce reprogramming.The dbDNA-iPSC cell line was identified through morphological analysis,alkaline phosphatase stai-ning,RT-PCR,and detection of pluripotent stem cell markers.The ZFN and TALEN gene editing techniques were employed to insert an 11.5 kb ultra-long DNA fragment containing the Cas9 expression cassette,EGFP reading frame with stop codons in the sequence,a eukaryotic cell screening marker and other elements at the iPSC AAVS1.The identification of the iPSC lines with Cas9 integrated into AAVS1 was conducted through the utilization of red fluorescence detection,genomic PCR reaction and Western blot.The EGFP single-strand homologous repair template and EGFP sgRNA were co-transfected into positive iPSC cell lines to repair the erroneous stop codons in the EGFP gene and to express green fluorescent protein correctly.This approach was undertaken in order to verify the gene editing ability of the Cas9 protein.Results The dbDNA vector was successfully pre-pared and employed for gene expression.The GFP gene expression efficiency of dbDNA was similar to that of traditional plas-mid DNA.The expression level of IFN-γ mRNA in the dbDNA group was lower than that of traditional plasmid DNA group.The iPSC cell lines resembling human embryonic stem cells were successfully obtained,iPSC cell lines with positive alka-line phosphatase staining,pluripotency related gene expression and stem cell pluripotency markers.The ultra-long fragment CRISPR-Cas9 gene editing reporting system was successfully inserted into the safe harbor AAVS1 site of the cell line,and the integrated fragment was successfully inserted into the genome by genomic PCR.Red fluorescent protein was expressed.Cas9 protein expression and EGFP gene recovery was confirmed by Western blot.Green fluorescence was observed.Conclusion A simple technical procedure for the preparation of dbDNA is successfully established.The gene expression efficiency of the dbD-NA vector is comparable to that of traditional plasmid DNA,with less immune response.The dbDNA-derived iPSCs are success-fully obtained,which is confirmed by morphological analysis,alkaline phosphatase staining,RT-PCR,and stem cell pluripotency markers.An 11.5 kb ultra-long DNA fragment is successfully inserted into the iPSC AAVS1 safe harbor to establish a stable iPSC cell line with Cas9-mediated gene editing.

Objective To establish a simple technical procedure for preparing doggybone DNA(dbDNA),and to discuss the distinctions in gene expression and immune response between dbDNA and conventional plasmid DNA.Induced pluripotent stem cell(iPSC)was generated from human peripheral blood mononuclear cells(PBMCs)by using dbDNA.To investigate a protocol for inserting ultra-long DNA fragments into the iPSC adeno-associated virus site 1(AAVS1),concurrently with the integration of the Cas9 expression frame point into the dbDNA-derived iPSC AAVS1 safe harbor,so as to establish a stable cell line that can achieve iPSC gene editing exclusively through the introduction of the single guide RNA(sgRNA)for the targeted gene.Methods A simple dbDNA preparation technique was established through utilization of Phi29 DNA polymerase multiple displacement amplification,protelomerase TelN digestion and covalent ligation,as well as the subsequent removal of the back-bone loop through the action of a restriction enzyme and exonuclease.The efficiency of green fluorescent protein(GFP)was eval-uated following electroporation of the dbDNA-GFP vector and the pMax-dbDNA-GFP traditional plasmid vector into PB-MCs.The mRNA expression of interferon-γ(IFN-γ)in the two groups was quantified by qPCR.The dbDNA reprogramming vectors dbDNA-MOS,dbDNA-KLF4,dbDNA-MYC and dbDNA-BCL were prepared and subsequently transferred into PBMCs to induce reprogramming.The dbDNA-iPSC cell line was identified through morphological analysis,alkaline phosphatase stai-ning,RT-PCR,and detection of pluripotent stem cell markers.The ZFN and TALEN gene editing techniques were employed to insert an 11.5 kb ultra-long DNA fragment containing the Cas9 expression cassette,EGFP reading frame with stop codons in the sequence,a eukaryotic cell screening marker and other elements at the iPSC AAVS1.The identification of the iPSC lines with Cas9 integrated into AAVS1 was conducted through the utilization of red fluorescence detection,genomic PCR reaction and Western blot.The EGFP single-strand homologous repair template and EGFP sgRNA were co-transfected into positive iPSC cell lines to repair the erroneous stop codons in the EGFP gene and to express green fluorescent protein correctly.This approach was undertaken in order to verify the gene editing ability of the Cas9 protein.Results The dbDNA vector was successfully pre-pared and employed for gene expression.The GFP gene expression efficiency of dbDNA was similar to that of traditional plas-mid DNA.The expression level of IFN-γ mRNA in the dbDNA group was lower than that of traditional plasmid DNA group.The iPSC cell lines resembling human embryonic stem cells were successfully obtained,iPSC cell lines with positive alka-line phosphatase staining,pluripotency related gene expression and stem cell pluripotency markers.The ultra-long fragment CRISPR-Cas9 gene editing reporting system was successfully inserted into the safe harbor AAVS1 site of the cell line,and the integrated fragment was successfully inserted into the genome by genomic PCR.Red fluorescent protein was expressed.Cas9 protein expression and EGFP gene recovery was confirmed by Western blot.Green fluorescence was observed.Conclusion A simple technical procedure for the preparation of dbDNA is successfully established.The gene expression efficiency of the dbD-NA vector is comparable to that of traditional plasmid DNA,with less immune response.The dbDNA-derived iPSCs are success-fully obtained,which is confirmed by morphological analysis,alkaline phosphatase staining,RT-PCR,and stem cell pluripotency markers.An 11.5 kb ultra-long DNA fragment is successfully inserted into the iPSC AAVS1 safe harbor to establish a stable iPSC cell line with Cas9-mediated gene editing.

Vascular calcification is a highly regulated process of ectopic calcification in cardiovascular system while no effective intervention can be clinically performed up to date.As vascular calcification undergoes a common regulatory mechanism within bone formation,bone morphogenetic protein 7(BMP-7)main-tains contractile phenotype of vascular smooth muscle cells and further inhibits vascular calcification via promoting the process of osteoblast differentiation,reducing ectopic calcification pressure by increasing bone formation and reducing bone resorption.This work systematically reviews the role of BMP-7 in vascular calcifi-cation and the possible mechanism,and their current clinical application as well.The current proceedings may help develope early diagnostic strategy and therapeutic treatment with BMP-7 as a new molecular marker and potential drug target.The expec-tation could achieve early prevention and intervention of vascular calcification and improve poor prognosis on patients.

Objective To establish a method to identify an unknown substance based on the combined use of gas chromatography-quadrupole time-of-flight mass spectrometry(GC-QTOF-MS),ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q/Orbitrap HRMS)and nuclear magnetic resonance(NMR)techniques.Methods The unknown substance was dissolved in methanol and was detected by GC-QTOF-MS and UPLC-Q/Orbitrap HRMS,and was dissolved in methanol-d4 to be detected by NMR.Results The main characteristics ion peaks of components with retention time of 9.67 min in GC-QTOF-MS measured were 84.080 8,110.999 7,128.107 0(base peak),138.994 7,etc.The protonated molecular ion peak m/z in UPLC-Q/Orbitrap HRMS was 268.109 3.It was inferred that the unknown substance was an analog of the syn-thetic cathinone substance 2-methyl-1-[4-(methylthio)phenyl]-2-morpholinopropan-1-one(MTMP)by comparing the mass spectrum information and molecular structure of MTMP.NMR analysis confirmed it as a novel N-morpholine substituted synthetic cathinone substance 1-(4-chlorophenyl)-2-methyl-2-morpholinopropan-1-one(CMMP).Conclusion The method established in this study can be used for structural confirmation of CMMP.

Idiosyncratic drug-induced liver injury (IDILI) has long posed a challenging and pivotal concern in pharmaceutical research. The complex composition of traditional Chinese medicine (TCM) has introduced a bottleneck in current research, hindering the elucidation of the component basis associated with IDILI in TCM. Using Epimedii Folium (EF) and Psoraleae Fructus (PF) as illustrative examples, this study endeavors to establish an in vitro evaluation model, providing a high-throughput and preliminary assessment method for screening components related to TCM-induced IDILI. A TNF-α-mediated HepG2 susceptible model was first established in this study, with the focus on the index components present in EF and PF. The release of lactate dehydrogenase (LDH) in the cell supernatant served as the detection index. A concentration-toxicity response curve was constructed, and the hepatotoxic components of EF and PF were identified utilizing the synergistic toxicity index. The LDH results unveiled the hepatotoxic effects of bavachin, backuchiol, isobavachin, neobavaisoflavone, psoralidin, isobavachalcone, icarisid I, and icarisid II on both normal and susceptible cells, categorizing these 8 components as both direct hepatotoxicity components and idiosyncratic hepatotoxicity components. Bavachin and neobavaisoflavone exhibited no hepatotoxicity on normal cells but demonstrated significant effects on susceptible cells, designating them as potential idiosyncratic susceptible hepatotoxicity components. The study further delineated that 10 EF components and 3 PF components were direct immune-promoting hepatotoxicity components. Additionally, 14 idiosyncratic immune-promoting hepatotoxicity components were identified, encompassing 10 EF components and 4 PF components, with neobavaisoflavone, bavachinin, and isobavachin being potential idiosyncratic susceptible immune-promoting hepatotoxicity components. Synergistic toxicity index results indicated that 13 idiosyncratic immune-promoting hepatotoxicity components (except anhydroicaritin) combined with bavachin demonstrated synergistic hepatotoxicity on susceptible cells. Notably, 3 idiosyncratic susceptible immune-promoting hepatotoxicity components combined with bavachin exhibited synergistic hepatotoxicity, with neobavaisoflavone displaying the highest synergistic toxicity index and bavachinin the lowest. In summary, this methodology successfully screens hepatotoxic and immune-promoting hepatotoxic components in EF and PF, distinguishing the types of components inducing hepatotoxicity, evaluating the hepatotoxicity degree of each component, and elucidating the synergistic relationships among them. Importantly, these findings align with the characteristics of IDILI. The method provides an effective model tool for the fundamental research of TCM-related IDILI components.

Objective: To analyze the clinical and molecular diagnostic status of Fanconi anemia (FA) in China. Methods: The General situation, clinical manifestations and chromosome breakage test and genetic test results of 107 pediatric FA cases registered in the Chinese Blood and Marrow Transplantation Registry Group (CBMTRG) and the Chinese Children Blood and Marrow Transplantation Registry Group (CCBMTRG) from August 2009 to January 2022 were analyzed retrospectively. Children with FANCA gene variants were divided into mild and severe groups based on the type of variant, and Wilcoxon-test was used to compare the phenotypic differences between groups. Results: Of the 176 registered FA patients, 69 (39.2%) cases were excluded due to lack of definitive genetic diagnosis results, and the remaining 107 children from 15 hospitals were included in the study, including 70 males and 37 females. The age at transplantation treatment were 6 (4, 9) years. The enrolled children were involved in 10 pathogenic genes, including 89 cases of FANCA gene, 7 cases of FANCG gene, 3 cases of FANCB gene, 2 cases of FANCE gene and 1 case each of FANCC, FANCD1, FANCD2, FANCF, FANCJ, and FANCN gene. Compound heterozygous or homozygous of loss-of-function variants account for 69.2% (72/104). Loss-of-function variants account for 79.2% (141/178) in FANCA gene variants, and 20.8% (37/178) were large exon deletions. Fifty-five children (51.4%) had chromosome breakage test records, with a positive rate of 81.8% (45/55). There were 172 congenital malformations in 80 children.Café-au-Lait spots (16.3%, 28/172), thumb deformities (16.3%,28/172), polydactyly (13.9%, 24/172), and short stature (12.2%, 21/172) were the most common congenital malformations in Chinese children with FA. No significant difference was found in the number of congenital malformations between children with severe (50 cases) and mild FANCA variants (26 cases) (Z=-1.33, P=0.185). Conclusions: FANCA gene is the main pathogenic gene in children with FA, where the detection of its exon deletion should be strengthened clinically. There were no phenotypic differences among children with different types of FANCA variants. Chromosome break test is helpful to determine the pathogenicity of variants, but its accuracy needs to be improved.
Male ; Female ; Humans ; Child ; Fanconi Anemia/genetics* ; Chromosome Breakage ; Retrospective Studies ; Exons ; China/epidemiology*

Objective: To examine the application value of 3D Slicer software assisted domestic frameless stereotactic robot in biopsy of intracranial lesions. Methods: A retrospective analysis was performed on 80 patients who admitted consecutively and underwent intracerebral lesions biopsy with the domestic frameless stereotactic robot at Department of Neurosurgery, Aerospace Central Hospital from January 2019 to December 2021. There were 36 males and 44 females, with a mean age of (38.5±18.0) years (range: 6 to 71 years). Before surgery only enhanced T1-weighted three-dimensional magnetization prepared gradient echo sequences and diffusion tensor imaging scans were performed. Self-reconstruction of intracranial lesions, cerebral cortex and blood vessels was carried out using 3D Slicer software system after the DICOM format imaging data of 80 patients were collected. These imaging data were merged to the workstation of the domestic frameless stereotactic robot for preoperative surgical planning and the surgical puncture path was designed to avoid blood vessels in the brain functional area, cerebral cortex and sulcus. Results: All frameless stereotactic biopsy were successfully performed. Postoperative pathological diagnosis included 50 cases of diffuse astrocytic and oligodendroglioma, 15 cases of lymphoma, 5 cases of metastatic tumors, 5 cases of inflammatory demyelinating disease, 2 cases of inflammatory granuloma, 1 case of hemangioma, 1 case of acute lymphoblastic leukemia intracranial invasion and 1 case of seminoma. The positive diagnosis rate was 100% (80/80). Postoperative imaging confirmed that the puncture path and target were accurately implemented according to the preoperative planning, and the target error was (1.32±0.44) mm (range: 0.55 to 1.99 mm). One case of puncture-related bleeding occurred at the target after surgery and improved after treatment. Conclusion: The three-dimensional multimodal images reconstructed by the 3D Slicer software before operation could help the surgeons make the preoperative planning and reduce the risk of stereotactic brain biopsy.
Male ; Female ; Humans ; Child ; Adolescent ; Young Adult ; Adult ; Middle Aged ; Aged ; Brain Neoplasms/pathology* ; Diffusion Tensor Imaging ; Retrospective Studies ; Robotics ; Biopsy ; Software ; Stereotaxic Techniques

Squamosa promoter binding protein-like (SPL) family is a group of important transcription factors involved in the regulation of plant growth and development and the response to environmental stress, but there are few studies in perennial fruit trees such as citrus. In this study, Ziyang Xiangcheng (Citrus junos Sib.ex Tanaka), an important rootstock of Citrus, was used as the material for analysis. Based on plantTFDB transcription factor database and sweet orange genome database, 15 SPL family members were genome-widely identified and cloned from Ziyang Xiangcheng, and named CjSPL1-CjSPL15. Sequence analysis showed that the open reading frame (ORF) length of CjSPLs ranged from 393 bp to 2 865 bp, encoding 130-954 amino acids. Phylogenetic tree divided 15 CjSPLs into 9 subfamilies. Gene structure and conserved domain analysis predicted 20 different conserved motifs and SBP basic domains. Analysis of cis-acting promoter elements predicted 20 different promoter elements, including those related to plant growth and development, abiotic stress and secondary metabolites. The expression patterns of CjSPLs under drought, salt and low temperature stresses were analyzed by real-time fluorescence quantitative PCR (qRT-PCR), and many CjSPLs were significantly up-regulated after stress treatment. This study provides a reference for further study on the function of SPL family transcription factors in citrus and other fruit trees.
Phylogeny ; Transcription Factors/metabolism* ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism* ; Multigene Family ; Stress, Physiological

Laminin subunit alpha 4 (LAMA4),a member of the laminin family,is present in the intercellular matrix of adult tissues as a major component of basement membrane.LAMA4 is involved in the adhesion of cells and can bind to corresponding integrins to activate relevant signaling pathways,playing an essential role in the growth,proliferation,and migration of cells.It has been demonstrated that LAMA4 is associated with the occurrence and development of a variety of diseases including tumors,and the expression of LAMA4 can be used as a biomarker of tumor diagnosis and prognosis.This paper summarizes the current research progress in LAMA4 with the focus on the relationship between LAMA4 and diseases,especially tumor,with a view to provide new directions for the future research.
Adult ; Humans ; Laminin ; Extracellular Matrix