Objective:To explore the correlation between CCAAT enhancer binding protein beta (CEBPB) expression and clinical characteristics in ccRCC, and to investigate the effect of CEBPB on proliferation and invasion of ccRCC cells.Methods:Between March 2020 to December 2020, the transcriptome and clinical data of 537 ccRCC cases were downloaded from TCGA database, and the correlation of CEBPB expression with clinical characteristics of ccRCC were analyzed. Univariate and multivariate Cox regression analysis were used to determine the effect of CEBPB expression on the prognosis of ccRCC patients. The correlation between CEBPB expression and immunocyte infiltration in ccRCC was investigated via TIMER database. The expression levels of CEBPB mRNA and protein in human renal tubular epithelial cell line HK2 and ccRCC cell lines (Caki-1, ACHN, 786O, 769P and A498) were determined by real-time PCR and western blot, respectively. After transfected with NC siRNA or CEBPB siRNA for 48 h, the proliferation and invasion of ACHN cells and 786O cells were determined by using MTT assay and invasion assay, respectively.Results:TCGA databases analysis revealed that, compared with normal kidney tissue, the expression of CEBPB mRNA in ccRCC was up-regulated by 2.55-fold ( P<0.05). CEBPB expression was positively correlated with age, tumor grade, tumor stage, lymph node metastasis and distant metastasis ( P<0.05). The tumor grade ( HR=1.703, P=0.040), tumor stage( HR=1.773, P=0.026), distant metastasis ( HR=3.080, P<0.001) and the high expression of CEBPB ( HR=1.874, P=0.003) were independent poor prognostic factors for ccRCC patients. The analysis results by using TIMER database showed that CEBPB expression was positively correlated with infiltrating levels of B cells (Rho=0.168), M2 macrophages (Rho=0.373), Tregs (Rho=0.348), neutrophils (Rho=0.194), and natural killer T cell (Rho=0.421) in ccRCC. The expression level of CEBPB mRNA in Caki-1, ACHN, 786O, 769P and A498 cells was (9.43±1.25)-fold, (5.44±0.82)-fold, (4.50±0.52)-fold, (4.88±0.73)-fold and (7.50 ± 1.04)-fold of HK2 cells, respectively. The expression level of CEBPB protein was (6.22±0.45)-fold, (5.84±0.85)-fold, (6.51±0.55)-fold, (6.23±0.62)-fold and (3.84±0.45)-fold of HK2 cells, respectively ( P<0.05). MTT assay showed that the proliferation rates of ACHN cells and 786O cells at 24, 48, 72, 96 h were (98.4±1.7)% and (99.0±1.4)%, (97.8±2.1)% and (98.5±1.5)%, (101.3±1.2)% and (97.6±1.7)%, (97.5±2.0)% and (99.1±1.3)% in NC siRNA group, and (68.8±5.8)% and (79.5±6.2)%, (57.9 ± 6.1)% and (70.8±5.1)%, (50.9±4.6)% and (66.8±4.9)%, (43.2±5.0)% and (60.5±5.3)% in CEBPB siRNA group. Compared with NC siRNA group, the proliferation activity of ACHN cells and 786O cells was significantly inhibited in the CEBPB siRNA group ( P<0.05). Cell invasion assay showed that the invasion activity of ACHN cells and 786O cells were (95.0±5.2)% and (97.3±4.4)% in NC siRNA group, (35.2±5.4)% and (26.7±3.3)% in CEBPB siRNA group, respectively ( P<0.05). Compared with NC siRNA group, the invasion activity of ACHN cells and 786O cells were significantly inhibited in the CEBPB siRNA group ( P<0.05). Conclusions:CEBPB was highly expressed in ccRCC, which was closely related to the prognosis and immunocyte infiltration of ccRCC patients. Silencing the expression of CEBPB significantly inhibited the proliferation and invasion of ccRCC cells

Lumbar spondylolysis refers to the bone injury between the upper and lower articular processes and the transition zone of the transverse process of the unilateral or bilateral pedicle of the lumbar spine, being a common cause of low back pain in patients that seriously affects their quality of life. The mechanism of the occurrence and development of lumbar spondylolysis is complex, and long-term stress wear and sudden damage with an external force are the main causes. At the same time, risk factors related to spinal anatomy are important causes of lumbar spondylolysis. A full understanding of the pathogenesis of lumbar spondylolysis, early identification of high-risk groups, and active preventive measures can reduce its incidence. For this purpose, the authors reviewed the research progress in risk factors related to the spinal anatomy of lumbar spondylolysis from three aspects including genetical susceptibility, local anatomy and overall spine-pelvic sequence, so as to provide references for the prevention and treatment of spondylolysis.

Objective To investigate the impact of LncRNA FOXP4-AS1 regulating the EZH2/LATS2 axis on the proliferation and migration of bladder urothelial carcinoma(BUC)cells.Methods The Cancer Genome Atlas(TCGA)database and real-time quantitative PCR were utilized to analyze the expression of FOXP4-AS1 and LATS2 mRNA in BUC tissues.Cell proliferation was observed using MTT experiments,and migration was checked using Transwell assays.Western blot assays were performed to determine the expression of LATS2 and H3K27me3 proteins.RNA immunoprecipitation(RIP)and chromatin immunoprecipitation(ChIP)assays were employed to verify the relationship between FOXP4-AS1,EZH2 and LATS2.Results Compared with normal bladder tissues,FOXP4-AS1 expression was increased in tumor tissues,while LATS2 mRNA expression was decreased(P<0.01).Moreover,FOXP4-AS1 expression was elevated in EJ,T24,BIU-87,and 5637 cell lines compared to SV-HUC-1(P<0.01).The inhibition of FOXP4-AS1 resulted in a significant decrease in the proliferation,migration,and expression of H3K27me3 protein in BUC cells,while concurrently upregulating the expression of LATS2 mRNA and protein.Conversely,the overexpression of FOXP4-AS1 yielded contrasting effects(P<0.05).RIP and ChIP assays revealed that FOXP4-AS1 could recruit EZH2 to the promoter region of LATS2,leading to an enrich-ment of H3K27me3 in this region.Interference with LATS2 or EZH2 expression partially reversed the effects of FOXP4-AS1 silencing or overexpression on the proliferation and migration of BUC cells,with concomitant effects on LATS2 expression.Conclusion FOXP4-AS1 demonstrates a notable increase in expression in BUC,leading to a suppression of LATS2 expression through the recruitment of EZH2 to the promoter region of LATS2.This regula-tory process ultimately influences the proliferation and migration of BUC cells.

Abstract: To explore the molecular mechanism by which long non-coding RNA Surfactant Associated 1 Pseudogene(SFTA1P) promotes the proliferation and migration of clear cell renal cell carcinoma(ccRCC) cells by regulating the microRNA-182-5p(miR-182-5p)/fibronectin 1(FN1) pathway. Methods: GEPIA2 software was utilized to analyze the expression ofSFTA1Pin ccRCC tissues from the TCGA database. Quantitative real-time PCR(qPCR) was employed to detect the expression ofSFTA1Pin ccRCC tissues, normal kidney tissues and ccRCC cell lines. A subcellular localization experiment was performed to explore the localization ofSFTA1Pwithin the human renal cell adenocarcinoma cell line(ACHN) derived from ccRCC. ACHN cells were then divided into the following groups: si-Con group, si-SFTA1P #2 group, mimic NC group, miR-182-5p mimic group, anti-miR-Con group, anti-miR-182-5p group, anti-miR-182-5p+si-FN1 group, si-Con+anti-miR-Con group, si-SFTA1P #2+anti-miR-Con group, and si-SFTA1P #2+anti-miR-182-5p group. CCK-8 and Transwell chamber experiments were conducted to assess cell proliferation and migration abilities. qPCR, Western blot, and dual-luciferase reporter assays were employed to elucidate the regulatory interactions amongSFTA1P,miR-182-5p, andFN1. Results: Analysis of The Cancer Genome Atlas(TCGA) database indicated thatSFTA1Pwas overexpressed in ccRCC tissues(P<0.05). When compared to normal kidney tissues,SFTA1Pexpression was markedly elevated in ccRCC tissues(P<0.01). Furthermore, the expression levels ofSFTA1Pin ccRCC cell lines 786-O, SN12-PM6, ACHN, and A498 were significantly higher than those in human renal proximal tubule cells(HK-2)(allP<0.01). Subcellular localization experiments revealed thatSFTA1Ppredominantly localized in the cytoplasm of ACHN cells. Compared to the si-Con group, the si-SFTA1P #2 group exhibited a significant reduction in proliferation and migration abilities of ACHN cells, accompanied by a decrease inFN1mRNA and protein expression(P<0.05). Compared to the mimic NC group, the expression ofFN1mRNA and protein in ACHN cells in the miR-182-5p mimic group reduced(P<0.01). In comparison to the anti-miR-Con group, the expression levels ofFN1mRNA and protein in ACHN cells were significantly elevated in the anti-miR-182-5p group. Additionally, there was a significant enhancement in both cell proliferation and migration capabilities(P<0.05). Conversely, the proliferation and migration abilities of ACHN cells in the anti-miR-182-5p+si-FN1 group were significantly reduced compared to the anti-miR-182-5p group(P<0.05). Furthermore, relative to the si-SFTA1P #2+anti-miR-Con group, the ACHN cells in the si-SFTA1P #2+anti-miR-182-5p group demonstrated increased proliferation and migration abilities, along with elevatedFN1mRNA and protein expression levels(P<0.05). Conclusion SFTA1Pexhibits elevated expression levels in ccRCC and facilitates the proliferation and migration of ccRCC cells through the modulation of themiR-182-5p/FN1signaling pathway.

Objective:To introduce a novel risk factor for lumbar spondylolysis, the pedicle isthmus angle (PIA), and to explore its underlying mechanism and clinical relevance.Methods:A retrospective analysis of CT imaging data from young male patients with lumbar spondylolysis, admitted to the 960th Hospital of the Joint Logistic Support Force of the PLA between January 2018 and August 2023, was conducted. The study included 119 cases of unilateral spondylolysis and 339 cases of bilateral spondylolysis, with a mean age of 22.8±3.4 years (range 18-30 years). A control group of 458 patients with normal lumbar CT scans, presenting with low back pain, was also analyzed. Their mean age was 22.9±3.5 years (range 18-30 years). The PIA of the left and right sides of the L 3, L 4, and L 5 vertebrae in both the spondylolysis and control groups were measured using CT imaging. Differences in PIA measurements between the left and right sides, as well as between groups, were compared. Binary logistic regression analysis identified risk factors for lumbar spondylolysis. The receiver operating characteristic (ROC) curve and Youden index were used to determine the critical risk threshold for lumbar spondylolysis. Results:No significant differences were found between the spondylolysis and control groups in terms of gender, age, height, weight, or body mass index (BMI) ( P>0.05). Similarly, there was no significant difference in the left and right PIA measurements for the L 3, L 4, and L 5 vertebrae in either group ( P>0.05). The PIA of the L 3 and L 4 vertebrae was not significantly different between the groups (107.2°±3.5° vs. 107.1°±3.5°, t=0.270, P=0.787; 110.6°±3.5° vs. 110.5°±4.0°, t=0.441, P=0.659). However, the PIA of the L 5 vertebra was significantly larger in the spondylolysis group (117.7°±4.7°) compared to the control group (114.0°±4.9°) ( t=11.654, P<0.001). Logistic regression analysis identified an increased PIA at L 5 ( β=0.159, OR=1.172, P<0.001) as a risk factor for lumbar spondylolysis. According to the ROC curve and Youden index, the risk of lumbar spondylolysis increased substantially when the L 5 PIA exceeded 115.8°. The area under the curve (AUC) was 0.709, with a sensitivity of 0.670 and a specificity of 0.644. Conclusion:PIA is an objective and effective imaging parameter for predicting lumbar spondylolysis. It aids in understanding the pathophysiology of spondylolysis, identifying high-risk individuals, and informing prevention and treatment strategies for lumbar spondylolysis.

OBJECTIVETo estimate the value of blastocyst culture for preimplantation genetic diagnosis (PGD).

METHODSDay 3 embryos were biopsied and analyzed with fluorescence in situ hybridization (FISH) technique. Embryos with normal FISH results were cultured into blastocysts, and the ones with better morphology scores were transferred. Fourteen embryos with abnormal FISH results were cultured into blastocysts. Part of the cells taken from the blastocysts were amplified by whole genomic amplification (WGA) and assessed by array-based comparative genomic hybridization (array-CGH) analysis.

RESULTSSix blastocysts with normal FISH results were transferred in 5 cycles. Four healthy babies of 3 cycles were delivered. Another one was a singleton pregnancy but with embryo growth arrest, whose villus karyotype was normal. Fourteen embryos with abnormal FISH results were cultured into blastocysts and analyzed by array-CGH. Six blastocysts were normal by array-CGH.

CONCLUSIONFISH combined with blastocyst culture may further ensure the accuracy of PGD result. Detection at the blastocyst stage can avoid false positive results and mosaic interferences on Day 3 stage and are therefore more authentic.

Adult ; Blastocyst ; cytology ; Comparative Genomic Hybridization ; methods ; Embryo Transfer ; Female ; Genetic Diseases, Inborn ; diagnosis ; embryology ; genetics ; prevention & control ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Male ; Pregnancy ; Preimplantation Diagnosis ; methods

Objective: To study the correlation of lifestyle characteristics with thyroid nodules in a population-based sample of centenarians in Hainan. Methods: The study was based on China Hainan Centenarian Cohort Study (CHCCS) conducted in 18 cities and counties in Hainan province from 2014 to 2016. A group of multidisciplinary team interviewed and examined local centenarians with structured questionnaires and ultrasonography procedures. A total of 918 centenarians were analyzed after excluding those who refused ultrasonographic examinations or had relevant missing data. Thyroids of centenarians were examined by 3-year experienced sonographer, details on lifestyle characteristics and dietary habits were collected by standard procedure. Results: Of the 918 centenarians, 683 (74.4%) had thyroid nodules under the ultrasonography procedures. The prevalence of thyroid nodules in different group of areca nut consumption varied significantly (P<0.05). In multivariate logistic regression model, the results showed female and areca nut intake were independently associated with thyroid nodules in centenarians (P<0.05). Conclusions The modifiable variables including areca nut intake as well as non-modifiable such as gender are independent determinants of thyroid nodules in centenarians. Further studies are warranted to explore and verify these associations in populations with different ranges of age.

Objective:To analyse the clinical significance of selective single embryo transfer by time-lapse mo-nitoring(TLM)or conventional morphology assessment(CMA)in vitro fertilization/intracytoplasmic sperm in-jection and embryo transfer(IVF/ICSI-ET),and to initially explore the predictive value of Raman spectral analy-sis of embryo culture medium for clinical pregnancy rate.Methods:The study is a prospective randomized con-trolled clinical trial.We assigned 139 patients treated with IVF/ICSI-ET in Reproductive and Genetics Center of Suzhou Municipal Hospital from April 2019 to July 2020,which were randomly assigned to either the CMA or the TLM group.We performed selective single-embryo transfer(fresh cycle and FET)after selecting the optimal em-bryos with TLM or CMA respectively.If the patient's first embryo transfer was unsuccessful,a second one would be performed to compare the differences in the cumulative live birth rate of embryo transfer and other pregnancy outcomes between the two groups.Meanwhile,we collected 15 μl of embryo culture medium at day 3 after IVF/ISCI fertilization for Raman spectroscopy analysis.Results:There were no differences in cumulative live birth,cu-mulative clinical pregnancy,cumulative premature birth,cumulative early spontaneous abortion,cumulative ectopic pregnancy and LGA or SGA between TLM and CMA groups(P＞0.05).The Neonatal sex ratio in the TLM group was lower than that in the CMA group,but the difference was not significant(P＞0.05).Raman spectros-copy analysis of embryo culture medium predicted the clinical pregnancy rate with 67.21％accuracy.Conclu-sions:In young women with a good ovarian reserve,the advantage of using TLM to evaluate embryos is not obvi-ous,so we should remain vigilant that embryo selection based on morphokinetic parameters may affect the sex ratio.Raman spectroscopic analysis of embryo culture medium is not yet able to effectively predict the planting ability of embryos.

Postzygotic mutations are acquired in normal tissues throughout an individual's lifetime and hold clues for identifying mutagenic factors.Here,we investigated postzygotic mutation spectra of healthy individuals using optimized ultra-deep exome sequencing of the time-series samples from the same volunteer as well as the samples from different individuals.In blood,sperm,and muscle cells,we resolved three common types of mutational signatures.Signatures A and B represent clock-like mutational processes,and the polymorphisms of epigenetic regulation genes influence the pro-portion of signature B in mutation profiles.Notably,signature C,characterized by C＞T transitions at GpCpN sites,tends to be a feature of diverse normal tissues.Mutations of this type are likely to occur early during embryonic development,supported by their relatively high allelic frequencies,presence in multiple tissues,and decrease in occurrence with age.Almost none of the public datasets for tumors feature this signature,except for 19.6％of samples of clear cell renal cell carcinoma with increased activation of the hypoxia-inducible factor 1(HIF-1)signaling pathway.Moreover,the accumulation of signature C in the mutation profile was accelerated in a human embryonic stem cell line with drug-induced activation of HIF-1α.Thus,embryonic hypoxia may explain this novel signature across multiple normal tissues.Our study suggests that hypoxic condition in an early stage of embryonic development is a crucial factor inducing C＞T transitions at GpCpN sites;and indi-viduals'genetic background may also influence their postzygotic mutation profiles.

Bone organoids can simulate the complex structure and function of the bone tissues, which makes them a frontier technology in organoid researches. Bone organoids show a tremendous potential of applications in bone disease modeling, bone injury repair, and medicine screening. Although advancements have been made so far in constructing bone organoids with functional structures like mineralization, bone marrow, trabecular bone, callus, woven bone, etc, the researches in this field are confronted with numerous challenges such as lack of standardized construction strategies and unified evaluation criteria, which limits their further promotion and application. To standardize researches in bone organoids, the Orthopedic Expert Committee of Geriatric Branch of Chinese Association of Gerontology and Geriatrics, the Youth Osteoporosis Group of Orthopedic Branch of Chinese Medical Association, the Osteoporosis Group of Orthopedic Surgeon Branch of Chinese Medical Doctor Association, and the Osteoporosis Committee of Shanghai Association of Integrated Traditional Chinese and Western Medicine organized related experts to formulate Expert consensus on the construction, evaluation, and application of bone organoids ( version 2024) based on an evidence-based approach. A total of 17 recommendations were put forth, aiming to standardize researches and clinical applications of bone organoids and enhance their value in scientific research and clinical practice.