BACKGROUND: Severe preeclampsia (sPE) is a serious condition posing risks to both maternal and fetal health. Based on mass spectrometry analysis, we identified a key protein, PSME3 (proteasome activator subunit 3), an 11S proteasome activator, whose protein level was significantly downregulated in sPE placentas and whose function in sPE remains unknown. METHODS: PSME3 protein levels in human placental tissue were detected using Western blot, and PSME3 concentration in serum was detected by ELISA assay. The human preeclampsia-like phenotypes of Psme3 − / − pregnant mice were examined. Trophoblast cell apoptosis was detected by flow cytometry. Pregnant mice were treated with 9.5% O 2 to construct a preeclampsia mouse model for detecting placental Psme3 expression. The regulation of PSME3 expression by hypoxia was detected in trophoblast cell lines treated with 21% O 2 or 1% O 2 . RESULTS: PSME3 protein levels were significantly downregulated in sPE placentas and serum. Pregnant mice with Psme3 − / − embryos and placentas spontaneously presented human preeclampsia-like symptoms, including hypertension and proteinuria, increased serum soluble fms-like tyrosine kinase 1 concentration, fetal growth restriction, and increased cellular apoptosis. Mechanically, PSME3 knockdown promoted the apoptosis of trophoblast cells by repressing the degradation of UBE2V2 (ubiquitin conjugating enzyme E2 V2). Moreover, the placentas of hypoxia-induced preeclampsia mice presented significantly reduced Psme3 protein levels and elevated Ube2v2 protein levels. Hypoxia-inducible factor-1α functioned as a transcriptional repressor of PSME3. CONCLUSIONS: In sPE placentas, hypoxia of the placenta may lead to the transcriptional inhibition of PSME3. PSME3 deficiency promotes the accumulation of UBE2V2, thereby inducing trophoblast cell apoptosis. Our study provides a new perspective for elucidating the pathogenesis of sPE.

Abstract Background The relationship between systemic immune inflammation index (SII) and the osteoarthritis (OA) has not been well established. This study aimed to investigate the associations between SII and OA risk using data from the National Health and Nutrition Examination Survey (NHANES) 2005–2016. Methods A cross-sectional study was conducted using data from participants in NHANES 2005–2016. SII is calculated as platelet count × neutrophil count/lymphocyte count. Multivariable logistic regression models were used to explore the link between SII and osteoporosis, adjusting for potential confounders. Results A total of 18,459 participants were included, of which 1130 reported having OA. After adjusting for potential confounders, an inverse association was observed between SII and OA (OR = 1.27; 95% CI = 1.07–1.51, p = 0.006). Moreover, we also identify that SII of 515.56 is the optimal cut-off value, indicating that when the SII value exceeds the cut-off value, the risk increases rapidly. Conclusions In this large, nationally representative sample of U.S. adults, higher SII was negatively associated with the risk of OA. Further prospective studies are needed to confirm these findings and elucidate potential underlying mechanisms.

Abstract Background: The relationship between dietary niacin, vitamin B6, and folate intakes and the risk of rheumatoid arthritis (RA) has not been well established. This study aimed to investigate the associations between these nutrients and RA risk using data from the National Health and Nutrition Examination Survey (NHANES) 2003-2018. Methods: A cross-sectional study was conducted using data from participants in NHANES 2003-2018. Dietary niacin, vitamin B6, and folate intakes were assessed using 24-hour dietary recall interviews. The presence of RA was determined based on self-reported physician diagnoses. Multivariable logistic regression models were used to estimate the adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for RA risk according to the intake of niacin, vitamin B6, and folate, adjusting for potential confounders. Results: A total of 24,151 participants reported having RA. After adjusting for potential confounders, an inverse association was observed between dietary niacin intake and RA risk (OR for the highest versus lowest quartile: 0.67; 95% CI: 0.54-0.84; Ptrend <0.001). Similarly, an inverse association were detected between vitamin B6 (OR: 0.66; 95% CI: 0.54-0.80; Ptrend <0.001) or folate (OR: 0.78; 95% CI: 0.64-0.95; P trend <0.001) intake and RA risk. Conclusions: In this large, nationally representative sample of U.S. adults, dietary intake of niacin, vitamin B6, and folate was negatively associated with the risk of RA. Further prospective studies are needed to confirm these findings and elucidate potential underlying mechanisms.

Subclassification of tumors based on molecular features may facilitate therapeutic choice and increase the response rate of cancer patients. However, the highly complex cell origin involved in osteosarcoma (OS) limits the utility of traditional bulk RNA sequencing for OS subclassification. Single-cell RNA sequencing (scRNA-seq) holds great promise for identifying cell heterogeneity. However, this technique has rarely been used in the study of tumor subclassification. By analyzing scRNA-seq data for six conventional OS and nine cancellous bone (CB) samples, we identified 29 clusters in OS and CB samples and discovered three differentiation trajectories from the cancer stem cell (CSC)-like subset, which allowed us to classify OS samples into three groups. The classification model was further examined using the TARGET dataset. Each subgroup of OS had different prognoses and possible drug sensitivities, and OS cells in the three differentiation branches showed distinct interactions with other clusters in the OS microenvironment. In addition, we verified the classification model through IHC staining in 138 OS samples, revealing a worse prognosis for Group B patients. Furthermore, we describe the novel transcriptional program of CSCs and highlight the activation of EZH2 in CSCs of OS. These findings provide a novel subclassification method based on scRNA-seq and shed new light on the molecular features of CSCs in OS and may serve as valuable references for precision treatment for and therapeutic development in OS.

Background: The relationship between Mg (magnesium), Cu (copper), and K (potassium) intakes and the risk of rheumatoid arthritis (RA) remains limited. The aim of present study was to examine the associations between Mg, Cu and K intakes with RA. Methods: Using data from the National Health and Nutrition Examination Survey (NHANES) 2003-2018, we examined the association between Mg, Cu and K intakes and the risk of RA among US adults. After adjustment for age, sex, race, BMI, educational level, smoking history, alcohol consumption, family Poverty Income Ratio (PIR), diabetes and total daily energy intake, logistic regression models and smooth curve fitting were applied to examine the associations of Mg, Cu and K intakes with RA. Results: A total of 18,338 participants were included (1,008 participants with RA). The multivariate adjusted ORs (95% CI) of RA were [0.66 (0.51, 0.84)], [0.76 (0.60, 0.97)], and [0.75 (0.58, 0.97)] in the highest versus lowest quartile of magnesium intakes, respectively. A nonlinear association between Cu intakes and RA was found. When Cu intake (ln) was between 0.6-2.2 mg, the risk of RA reduced by 26% for every 1 mg increase of intake in Cu [0.74 (0.58, 0.96)]. Conclusions: Higher Mg, Cu and K intakes may be inversely associated with the risk of RA among US adults, and an inverse L-shaped association between dietary Cu and RA was found.

AbstractBackground As a natural process of aging, intervertebral disc degeneration is more prone to degeneration, with limited repair ability, which is closely related to ageing and excessive manual labor. The main manifestations include the formation of fissures in the annulus fibrosus, the reduction of water in the intervertebral disc, and the decrease of elasticity. However, the molecular mechanism of intervertebral disc degeneration is still unclear. Methods In this study, key biomarkers in intervertebral disc degeneration were identified through bioinformatics. GSE70362 and GSE56081 were downloaded from the GEO database, and then the two datasets were differentially analyzed and validated for their expression, immune infiltration analysis, functional enrichment analysis, and potential drug prediction through the Connectivity Map (CMap) database. Results A total of 352 and 9815 differential genes were identified by GSE70362, GSE56081, respectively. The up-regulated and down-regulated genes of the two datasets were intersected with ferroptosis genes to obtain five key genes that were significantly correlated with immune cell content, namely AKR1C3, CKB, KRT19, MT1G and MUC1. The ROC results showed that the five core genes could well predict the occurrence and development of the disease. In addition, the results of CMap suggested that four drugs, including 1-Phenylbiguanide, LY-2183240, Flubendazole and Penciclovir, have the potential to reverse intervertebral disc degeneration. Conclusion Exploring the expression levels of five key genes in intervertebral disc degeneration is conductive to providing new ideas for the prevention and treatment of intervertebral disc degeneration. Moreover, Flubendazole and Penciclovir have the potential to provide options for clinical treatment of intervertebral disc degeneration.

Background Neurofilament light chain (NEFL) has been identified as a biomarker for spinal cord injury (SCI), but its effect and underlying mechanism in SCI remain unclear. Methods SCI rat models were established for in vivo studies. Lipopolysaccharide (LPS)-induced cell models were used for in vitro studies. The protein and mRNA expression levels of genes were evaluated by western blotting and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The pathological changes in rats after SCI were subjected to histological examinations. The interaction of NEFL and upstream miRNAs was explored using dual-luciferase reporter gene assays. Results NEFL was highly expressed in SCI rat spinal cord tissues and LPS-stimulated PC12 cells. NEFL silencing showed an inhibitory effect on the morphological changes of SCI rats and the secretion of inflammatory factors and facilitated functional recovery of SCI rats. MiR-30b-5p was demonstrated to target NEFL and negatively regulate NEFL mRNA and protein levels. Downregulation of miR-30b-5p in SCI cell and rat models was demonstrated. MiR-30b-5p alleviated the inflammatory response in SCI rat models and LPS-stimulated PC12 cells and promoted functional recovery in rats by targeting NEFL. NEFL activated mTOR signaling. MiR-30b-5p inactivated mTOR signaling by negatively regulating NEFL. Conclusion MiR-30b-5p alleviated the inflammatory response and facilitated the functional recovery of SCI rats by targeting NEFL to inactivate the mTOR pathway.

Purpose . To investigate the relationship between serum cotinine and lumbar bone mineral density (BMD) among 7905 participants aged 30 years and over. Method . A total of 3945 men and 3960 women from the National Health and Nutrition Examination Survey 2011–2018 were included in this cross-sectional analysis. Independent variable was serum cotinine, which is a biomarker of cigarette exposure. The outcome variable was lumbar BMD. We investigated the associations of serum cotinine levels and lumbar BMD using multivariable linear regression models. Results . Serum cotinine concentration was negatively associated with lumbar BMD after adjustment of relevant covariables ( β  = −0.039, 95% CI: −0.078 to −0.014, P = 0.005). However, in the subgroup analysis stratified by gender, this negative association remained only in women ( β  = −0.072, 95% CI: −0.132 to −0.012, P = 0.019). Conclusion . Our study suggested that elevated serum cotinine level correlated with decreased lumbar BMD, especially in women. This finding indicated that reducing cigarette exposure and maintaining serum cotinine at a low level may be beneficial to bone health for adults.

目的 探讨杨浦区中老年人群腰椎骨量减少现状并分析相关因素,为制定针对性预防干预措施提供依据.方法 以2019年1月1日至2020年12月31日 在上海市某医院体检中心进行健康体检的≥45岁人群作为研究对象,对其进行问卷调查、血钙、碱性磷酸酶(ALP)及腰椎骨密度检测,从健康体检结果中提取体格检查及血生化检测结果,采用描述流行病学分析方法对腰椎骨量减少现状进行分析,并采用单、多因素分析方法对中老年人群腰椎骨量减少发生的影响因素进行分析.结果 本研究共获得1 826名杨浦区≥45岁健康体检者资料,男性占38.61％,女性占61.39％.年龄以45～75岁.1 826名中老年人群中,1186例腰椎骨量减少,减少检出率为64.95％.多因素Logistic回归分析结果发现年龄越大(OR=4.057)、有骨折史(OR=2.854)的≥45岁健康体检人群腰椎骨量减少可能性较大,ALP＞70 mmol/L(OR=0.325)、每天运动≥1 h(OR=0.337)的≥45岁健康体检人群腰椎骨量减少可能性较小.结论 中老年人群易受年龄、运动等因素影响,腰椎骨量减少检出率较高,实际生活中需通过多种途径强化运动、体重等方面管理,最大限度改善腰椎骨密度,降低骨质疏松发生风险.

Background Enhanced recovery after surgery (ERAS) pathway in spine surgery is increasingly popular which can reduce the length of hospital stay (LOS). However, there are few studies on the safety and effectiveness of ERAS pathway in the treatment of single-level lumbar disc herniation (LDH) by percutaneous endoscopic transforaminal discectomy (PETD). The aim of this study was to investigate whether ERAS can reduce LOS of patients with single segment LDH treated by PETD. Methods We reviewed the outcomes of all LDH patients (L4/5) who had been treated with PETD at our institution. Quasi-experimental study was adopted between patients treated in an ERAS after PETD with those rehabilitated on a traditional pathway. The two groups were analyzed for LOS, operation time, complications, visual analog scale (VAS), Oswestry Dysfunction Index (ODI), hospitalization expenses (HE), and improved MacNab efficacy assessment criteria (MacNab). Results A total of 120 single segment LDH patients (ERAS pathway 60 cases, traditional care pathway 60 cases) who were selected from January 2019 to January 2021 met the inclusion criteria. There was a significant difference in mean LOS postoperative VAS scores and ODI on the 3rd day after surgery between the two groups ( P < 0.05). The incidence of complications and HE were similar in the two groups ( P > 0.05). The mean LOS decreased from 3.47 ± 1.14 days to 5.65 ± 1.39 days after application of ERAS pathway ( P < 0.05). Conclusions The ERAS pathway reduced LOS without resulting in additional complications after PETD. These findings support the application of the perioperative ERAS pathway in the treatment of single-level LDH with PETD. Level of evidence Level IV, therapeutic

Leucine-rich repeat containing G protein-coupled receptor 5 (Lgr5), a marker of intestinal stem cells (ISCs), is considered to play key roles in tissue homoeostasis and regeneration after acute radiation injury. However, the activation of Lgr5 by integrated signaling pathways upon radiation remains poorly understood. Here, we show that irradiation of mice with whole-body depletion or conditional ablation of REGγ in Lgr5+ stem cell impairs proliferation of intestinal crypts, delaying regeneration of intestine epithelial cells. Mechanistically, REGγ enhances transcriptional activation of Lgr5 via the potentiation of both Wnt and Hippo signal pathways. TEAD4 alone or cooperates with TCF4, a transcription factor mediating Wnt signaling, to enhance the expression of Lgr5. Silencing TEAD4 drastically attenuated β-catenin/TCF4 dependent expression of Lgr5. Together, our study reveals how REGγ controls Lgr5 expression and expansion of Lgr5+ stem cells in the regeneration of intestinal epithelial cells. Thus, REGγ proteasome appears to be a potential therapeutic target for radiation-induced gastrointestinal disorders.

Amphiphysin 1 (AMPH-1) is a nerve terminals-enriched protein involved in endocytosis, and we observe that its expression is increased in breast cancer tumor in compared with normal breast. However, its function in breast cancer is unknown. Here we aim to explore the role of AMPH-1 in breast cancer cells. Knockdown of AMPH-1 in breast cancer cells promotes cell proliferation, cell cycle progression and cell migration, and attenuates cell apoptosis. Of note, knockdown of AMPH-1 promotes breast cancer progression in xenograft mouse model. These oncogenic phenotypes may be partially due to the activated EMT and ERK pathways after inhibition of AMPH-1. Oncomine analyses of multiple breast cancer patient datasets show that reduced AMPH-1 mRNA level is significantly associated with breast cancer patients having metastatic events, advanced stage, poor clinical outcomes, and Paclitaxel+FEC treatment resistance. In summary, our results identified the anti-oncogenic function of AMPH-1 in breast cancer in vitro and in vivo. Activation of AMPH-1 may be a promising approach to treat breast cancer patients.

Background: Tumor microenvironment, in particular the stroma, plays an important role in breast cancer cell invasion and metastasis. Investigation of the molecular characteristics of breast cancer stroma may reveal targets for future study. Methods: The transcriptome profiles of breast cancer stroma and normal breast stroma were compared to identify differentially expressed genes (DEGs). The method was analysis of GSE26910 and GSE10797 datasets. Common DEGs were identified and then analyses of enriched pathways and hub genes were performed. Results: A total of 146 DEGs were common to GSE26910 and GSE10797. The enriched pathways were associated with "extracellular matrix (ECM) organization", "ECM-receptor interaction" and "focal adhesion". Network analysis identified six key genes, including JUN, FOS, ATF3, STAT1, COL1A1 and FN1. Notably, COL1A1 and FN1 were identified for the first time as cancer stromal key genes associated with breast cancer invasion and metastasis. Oncome analysis showed that the high expression levels of COL1A1 and FN1 correlated to an advanced stage of breast cancer and poor clinical outcomes. Conclusions: We found that several conserved tumor stromal genes might regulate breast cancer invasion through ECM remodeling. The clinical outcome analyses of COL1A1 and FN1 suggest these two genes are promising targets for future studies.

The expression of microRNA-215 (miR-215) in non-small cell lung cancer (NSCLC) tissues and the effects of miR-215 on the proliferation and migration of NSCLC cells were investigated. qRT-PCR was used to detect the expression of miR-215 in NSCLC tissues and paired normal tumor-adjacent lung tissues; MTT assay, transwell assay and soft-agar assay were used in vitro to evaluate the role of miR-215 on proliferation, migration and cell clonality on NSCLC cells, after transfecting miR-215 mimics to NSCLC cell line A549 or miR-215 to H1299. miR-215 was significantly decreased in NSCLC tissues compared to the paired normal tissues; Overexpression of miR-215 in A549 cells resulted in reduction of the cell proliferation, migration and cell clonality, while downregulation of miR-215 in H1299 cells could promote cell proliferation, migration and clonality. In conclusion, miR-215 was downregulated in NSCLC tissues and may play a key role in the development of NSCLC.

目的:研究微小RNA-203表达与骨肉瘤细胞生物学特征的相关性.方法:收集骨肉瘤组织和肉瘤旁正常组织,检测miRNA-203的表达量;培养骨肉瘤细胞株MG63和成骨细胞细胞株hFOB1.19,检测细胞中miRNA-203的表达量;转染miRNA-203的模拟物后6h、12h、18h、24h时,检测细胞增殖、侵袭能力以及相关基因的表达量.结果:骨肉瘤组织中miRNA-203的表达量低于肉瘤旁正常组织,骨肉瘤细胞株MG63中miRNA-203的表达量低于成骨细胞细胞株hFOB1.19;在骨肉瘤细胞株MG63中,miRNA-203的模拟物能够以时间依赖性的方式降低细胞活力、减少侵袭细胞的数目,抑制Bcl-2、cMet、Notch-3、MMP-9的表达.结论:骨肉瘤组织中微小RNA-203的表达量显著降低,增加微小RNA-203的表达能够抑制骨肉瘤细胞的增殖和侵袭,其靶基因为Bcl-2、cMet、Notch-3、MMP-9.

目的：探讨行为回放式教学法在骨科物理诊断教学中的应用效果。方法：选择第二军医大学2012级五年制临床医学专业学员作为观察对象，其中实验组40名学员，采用行为回放式教学方法进行教学；对照组40名学员，采用传统教学模式进行教学。结果：实验组学员的操作能力考核成绩明显优于对照组，同时在学习兴趣、理论知识理解、解决实际问题能力和面对患者自信心四个方面明显优于对照组。结论：行为回放式教学法在骨科物理诊断教学中可行性强，可以显著提高临床教学效果和学员素质，值得提倡和推广。

Objective To evaluate the efficacy of clinical pathway teaching method applied in practice teaching of orthopedic oncology.Methods Sixty orthopedics graduates of grades 2011 to 2014 from Changzheng Hospital of the Second Military Medical University were randomized into the experimental group and the control group.Thirty students in the experimental group were taught with clinical pathway method,while the other 30 students in the control group were taught with the traditional teaching method.Results Significant improvements in therapeutic strategy examination were observed in the experimental group.Moreover,self-evaluation survey showed that understanding of theoretical knowledge,problem-solving ability and confidence in face of patients were all improved significantly in the experimental group.Conclusion Clinical pathway teaching method has great feasibility in orthopedic oncology practice teaching.It can obviously improve the teaching efficacy and students'quality.In this regard,it deserves promotion and popularization.

Purpose A number of studies have demonstrated that microRNAs play a critical role in osteosarcoma progression, therapy and drug resistance. MicroRNA-202 has proven to be dysregulated in many human cancer studies. This study aimed to explore miR-202 contributions to drug resistance in osteosarcoma. Methods miR-202 expression was measured by real-time PCR in patient tissues, cell lines or cells treated with TGF-β1, using siRNA to knock down the expression of Smad2, Smad3 and Smad4. miR-202 mimics, inhibitor and scramble siRNA were transfected into osteosarcoma cells to observe effects on cell apoptosis and drug resistance. Moreover, relationships of miR-202 level with PDCD4 were investigated by luciferase reporter assay, real-time PCR and Western blotting. Results We found that miR-202 is overexpressed in osteosarcoma tissues when compared with normal human osteoblasts and TGF-β1 can induce the expression of miR-202. Transfection of miR-202 mimics into osteosarcoma cell lines significantly promotes chemotherapy resistance by targeting PDCD4, a tumor suppressor which is involved in apoptosis. In contrast, transfection of miR-202 inhibitor enhances the drug sensitivity and apoptosis. Conclusions This study provides new insights into miRNA-202 in osteosarcoma as a potential molecular target for chemotherapy.