Potential anti-liver cancer targets and mechanisms of kaempferitrin based on network pharmacology, molecular docking and experimental verification.

AIM: Kaempferitrin is an active component in Chenopodium ambrosioides, showing medicinal functions against liver cancer. This study aimed to identify the potential targets and pathways of kaempferitrin against liver cancer using network pharmacology and molecular docking, and verify the essential hub targets and pathway in mice model of SMMC-7721 cells xenografted tumors and SMMC-7721 cells. METHODS: Kaempferitrin therapeutical targets were obtained by searching SwissTargetPrediction, PharmMapper, STITCH, DrugBank, and TTD databases. Liver cancer specific genes were obtained by searching GeneCards, DrugBank, TTD, OMIM, and DisGeNET databases. PPI network of "kaempferitrin-targets-liver cancer" was constructed to screen the hub targets. GO, KEGG pathway and MCODE clustering analyses were performed to identify possible enrichment of genes with specific biological subjects. Molecular docking and molecular dynamics simulation were employed to determine the docking pose, potential and stability of kaempferitrin with hub targets. The potential anti-liver cancer mechanisms of kaempferitrin, as predicted by network pharmacology analyses, were verified by in vitro and in vivo experiments. RESULTS: 228 kaempferitrin targets and 2186 liver cancer specific targets were identified, of which 50 targets were overlapped. 8 hub targets were identified through network topology analysis, and only SIRT1 and TP53 had a potent binding activity with kaempferitrin as indicated by molecular docking and molecular dynamics simulation. MCODE clustering analysis revealed the most significant functional module of PPI network including SIRT1 and TP53 was mainly related to cell apoptosis. GO and KEGG enrichment analyses suggested that kaempferitrin exerted therapeutic effects on liver cancer possibly by promoting apoptosis via p21/Bcl-2/Caspase 3 signaling pathway, which were confirmed by in vivo and in vitro experiments, such as HE staining of tumor tissues, CCK-8, qRT-PCR and Western blot. CONCLUSION: This study provided not only insight into how kaempferitrin could act against liver cancer by identifying hub targets and their associated signaling pathways, but also experimental evidence for the clinical use of kaempferitrin in liver cancer treatment.

Irisin alleviated the reproductive endocrinal disorders of PCOS mice accompanied by changes in gut microbiota and metabolomic characteristics.

Introduction: Folliculogenesis and oligo/anovulation are common pathophysiological characteristics in polycystic ovary syndrome (PCOS) patients, and it is also accompanied by gut microbiota dysbiosis. It is known that physical activity has beneficial effects on improving metabolism and promoting ovulation and menstrual cycle disorder in PCOS patients, and it can also modulate the gastrointestinal microbiota in human beings. However, the mechanism remains vague. Irisin, a novel myokine, plays a positive role in the mediating effects of physical activity. Methods: Mice were randomly divided into the control group, PCOS group and PCOS+irisin group. PCOS model was induced by dehydroepiandrosterone (DHEA) and high-fat diet (HFD). The PCOS+irisin group was given irisin 400µg/kg intraperitoneal injection every other day for 21 days. The serum sex hormones were measured by radioimmunoassay. Hematoxylin and Eosin (H&E) Staining and immunohistochemistry (IHC) were conducted on ovarian tissue. The feces microbiota and metabolomic characteristics were collected by 16S rRNA gene sequencing and liquid chromatography-mass spectrometry (LC-MS). Results: In this study, we demonstrated that irisin supplementation alleviated reproductive endocrine disorders of PCOS mice, including estrous cycle disturbance, ovarian polycystic degeneration, and hyperandrogenemia. Irisin also improved the PCOS follicles dysplasia and ovulation disorders, while it had no significant effect on the quality of oocytes. Moreover, irisin could mitigate the decreased bacteria of Odoribacter and the increased bacteria of Eisenbergiella and Dubosiella in PCOS mice model. Moreover, irisin could alleviate the increased fecal metabolites: Methallenestril and PS (22:5(4Z,7Z,10Z,13Z,16Z)/ LTE4). Conclusion: These results suggest that irisin may alleviate the status of PCOS mice model by modulating androgen-induced gut microbiota dysbiosis and fecal metabolites. Hence, our study provided evidence that irisin may be considered as a promising strategy for the treatment of PCOS.

Integrating plasma protein-centric multi-omics to identify potential therapeutic targets for pancreatic cancer.

BACKGROUND: Deciphering the role of plasma proteins in pancreatic cancer (PC) susceptibility can aid in identifying novel targets for diagnosis and treatment. METHODS: We examined the relationship between genetically determined levels of plasma proteins and PC through a systemic proteome-wide Mendelian randomization (MR) analysis utilizing cis-pQTLs from multiple centers. Rigorous sensitivity analyses, colocalization, reverse MR, replications with varying instrumental variable selections and additional datasets, as well as subsequent meta-analysis, were utilized to confirm the robustness of significant findings. The causative effect of corresponding protein-coding genes' expression and their expression pattern in single-cell types were then investigated. Enrichment analysis, between-protein interaction and causation, knock-out mice models, and mediation analysis with established PC risk factors were applied to indicate the pathogenetic pathways. These candidate targets were ultimately prioritized upon druggability and potential side effects predicted by a phenome-wide MR. RESULTS: Twenty-one PC-related circulating proteins were identified in the exploratory phase with no evidence for horizontal pleiotropy or reverse causation. Of these, 11 were confirmed in a meta-analysis integrating external validations. The causality at a transcription level was repeated for neutrophil elastase, hydroxyacylglutathione hydrolase, lipase member N, protein disulfide-isomerase A5, xyloside xylosyltransferase 1. The carbohydrate sulfotransferase 11 and histo-blood group ABO system transferase exhibited high-support genetic colocalization evidence and were found to affect PC carcinogenesis partially through modulating body mass index and type 2 diabetes, respectively. Approved drugs have been established for eight candidate targets, which could potentially be repurposed for PC therapies. The phenome-wide investigation revealed 12 proteins associated with 51 non-PC traits, and interference on protein disulfide-isomerase A5 and cystatin-D would increase the risk of other malignancies. CONCLUSIONS: By employing comprehensive methodologies, this study demonstrated a genetic predisposition linking 21 circulating proteins to PC risk. Our findings shed new light on the PC etiology and highlighted potential targets as priorities for future efforts in early diagnosis and therapeutic strategies of PC.

Deep Learning Model Coupling Wearable Bioelectric and Mechanical Sensors for Refined Muscle Strength Assessment.

Muscle strength (MS) is related to our neural and muscle systems, essential for clinical diagnosis and rehabilitation evaluation. Although emerging wearable technology seems promising for MS assessment, problems still exist, including inaccuracy, spatiotemporal differences, and analyzing methods. In this study, we propose a wearable device consisting of myoelectric and strain sensors, synchronously acquiring surface electromyography and mechanical signals at the same spot during muscle activities, and then employ a deep learning model based on temporal convolutional network (TCN) + Transformer (Tcnformer), achieving accurate grading and prediction of MS. Moreover, by combining with deep clustering, named Tcnformer deep cluster (TDC), we further obtain a 25-level classification for MS assessment, refining the conventional 5 levels. Quantification and validation showcase a patient's postoperative recovery from level 3.2 to level 3.6 in the first few days after surgery. We anticipate that this system will importantly advance precise MS assessment, potentially improving relevant clinical diagnosis and rehabilitation outcomes.

Preoperative and follow-up variations of psoas major muscle are related to S1 screw loosening in patients with degenerative lumbar spinal stenosis.

BACKGROUND: It was reported the paraspinal muscle played an important role in spinal stability. The preoperative paraspinal muscle was related to S1 screw loosening. But the relationship between preoperative and postoperative change of psoas major muscle (PS) and S1 pedicle screw loosening in degenerative lumbar spinal stenosis (DLSS) patients has not been reported. This study investigated the effects of preoperative and follow-up variations in the psoas major muscle (PS) on the first sacral vertebra (S1) screw loosening in patients with DLSS. METHODS: 212 patients with DLSS who underwent lumbar surgery were included. The patients were divided into the S1 screw loosening group and the S1 screw non-loosening group. Muscle parameters were measured preoperatively and at last follow-up magnetic resonance imaging. A logistic regression analysis was performed to investigate the risk factors for S1 screw loosening. RESULTS: The S1 screw loosening rate was 36.32% (77/212). The relative total cross-sectional areas and relative functional cross-sectional areas (rfCSAs) of the PS at L2-S1 were significantly higher after surgery. The increased rfCSA values of the PS at L3-S1 in the S1 screw non-loosening group were significantly higher than those in the S1 screw loosening group. The regression analysis showed male, lower CT value of L1 and longer segment fusion were independent risk factors for S1 screw loosening, and postoperative hypertrophy of the PS was a protective factor for S1 screw loosening. CONCLUSIONS: Compared to the preoperative muscle, the PS size increased and fatty infiltration decreased after surgery from L2-3 to L5-S1 in patients with DLSS after short-segment lumbar fusion surgery. Postoperative hypertrophy of the PS might be considered as a protective factor for S1 screw loosening. MRI morphometric parameters and postoperative selected exercise of PS for DLSS patients after posterior lumbar fusion surgery might contribute to improvement of surgical outcome.

Impact of pelvic anteversion on spinopelvic alignment in an asymptomatic population: a dynamic perspective of standing and sitting.

BACKGROUND CONTEXT: A subgroup of patients with pelvic anteversion can present with an unusually large degree of lumbar lordosis (LL), a highly sloped sacrum, and a relatively small pelvic incidence (PI). Prior to lumbar surgery, it can be important to consider such unique sagittal alignment. However, until now, there has been a lack of a predictive model considering different pelvic alignments. Furthermore, the dynamic characteristics of an anteverted pelvis (AP) subgroup have also been unclear. PURPOSE: To build linear predictive formulas for LL that take pelvic anteversion into consideration and to explore the dynamic characteristics of an AP subgroup. STUDY DESIGN: Monocentric, cross-sectional study. PATIENT SAMPLE: Five hundred and sixty-five asymptomatic Chinese men and women between the ages of 18 and 80 years. OUTCOME MEASURES: Sagittal parameters including LL, lumbar lordosis minus thoracic kyphosis (LL-TK), PI, pelvic tilt (PT), pelvic incidence minus lumbar lordosis (PI-LL), sacral slope (SS), sacral slope divided by pelvic incidence (SS/PI), sagittal vertical axis (SVA), thoracic kyphosis (TK), and T1 (first thoracic vertebra) pelvic angle (TPA) were measured on whole spine radiographs obtained with participants in standing and sitting positions. METHODS: All participants underwent radiography in the standing position; 235 of them underwent additional radiography in the sitting position to allow measurement of sagittal parameters. The participants with pelvic anteversion were placed in an AP (anteverted pelvis) group. Sagittal parameters were compared between the AP group and the non-AP group, and predictive formulas for LL based on PI were created in both groups. In addition, changes in sagittal parameters from standing to sitting were compared in the AP group and a PI-matched control group. RESULTS: Of the 565 participants, 171 (30.3%) had pelvic anteversion. In comparison with the non-AP group, the AP group presented with larger LL, a larger SS, and a smaller PT, with relatively small PI. The predictive formulas for LL were LL=0.60° × PI+21.60° (R2=0.268; p<.001) in the whole cohort, LL=0. 83×PI+18.75° (R2=0.427; p<.001) in AP group, and LL=0.79°×PI+9.66° (R2=0.451; p<.001) in the non-AP group. In moving from standing to sitting, the AP group presented with a larger decrease in SS and LL compared with the control group, indicating different patterns of spinopelvic motion. CONCLUSIONS: In the cohort examined, 30.3% present with pelvic anteversion. Those with AP present with unique characteristics of spinopelvic alignment. In moving from standing to sitting, they exhibit different patterns of spinopelvic motion. We found that identifying the degree of anteversion in each person improves the accuracy of linear models for predicting the degree of LL, which in turn can make plans for spine surgery more accurate.

Konjac glucomannan-based highly antibacterial active films loaded with thyme essential oil through bacterial cellulose nanofibers/Ag nanoparticles stabilized Pickering emulsions.

The aim of this study was to develop novel konjac glucomannan (KGM)-based highly antibacterial active films, where five types of films were prepared and compared. The microstructure results showed that KGM-based films loaded with thyme essential oil (TEO) through bacterial cellulose nanofibers/Ag nanoparticles (BCNs/Ag nanoparticles) stabilized Pickering emulsions (Type V films) displayed the smoothest surface and the most evenly dispersed TEO droplets as compared with the other four types of films. Moreover, Type V films showed the highest contact angle value (86.28°), the best thermal stability and mechanical properties. Furthermore, Type V films presented the highest total phenol content (13.23 mg gallic acid equivalent/g film) and the best antioxidant activity (33.96 %) as well as the best sustained-release property, thus showing the best antibacterial activity, which was probably due to that BCNs/Ag nanoparticles and TEO displayed a synergistic effect to some extent. Consequently, Type V film-forming solutions were used as coatings for tangerines. The results showed that the tangerines treated with Type V coatings displayed excellent fresh-keeping properties. Therefore, the coatings, KGM-based film-forming solutions loaded with TEO through BCNs/Ag nanoparticles stabilized Pickering emulsions, have great potential for the preservation of fruits and vegetables.

Inhibition the ubiquitination of ENaC and Na,K-ATPase with erythropoietin promotes alveolar fluid clearance in sepsis-induced acute respiratory distress syndrome.

Sepsis-induced acute respiratory distress syndrome (ARDS) causes significant fatalities worldwide and lacks pharmacological intervention. Alveolar fluid clearance (AFC) plays a pivotal role in the remission of ARDS and is markedly impaired in the pathogenesis of ARDS. Here, we demonstrated that erythropoietin could effectively ameliorate lung injury manifestations and lethality, restore lung function and promote AFC in a rat model of lipopolysaccharide (LPS)-induced ARDS. Moreover, it was proven that EPO-induced restoration of AFC occurs through triggering the total protein expression of ENaC and Na,K-ATPase channels, enhancing their protein abundance in the membrane, and suppressing their ubiquitination for degeneration. Mechanistically, the data indicated the possible involvement of EPOR/JAK2/STAT3/SGK1/Nedd4-2 signaling in this process, and the pharmacological inhibition of the pathway markedly eliminated the stimulating effects of EPO on ENaC and Na,K-ATPase, and subsequently reversed the augmentation of AFC by EPO. Consistently, in vitro studies of alveolar epithelial cells paralleled with that EPO upregulated the expression of ENaC and Na,K-ATPase, and patch-clamp studies further demonstrated that EPO substantially strengthened sodium ion currents. Collectively, EPO could effectively promote AFC by improving ENaC and Na,K-ATPase protein expression and abundance in the membrane, dependent on inhibition of ENaC and Na,K-ATPase ubiquitination, and resulting in diminishing LPS-associated lung injuries.

Relationship between the postoperative variations of paraspinal muscles and adjacent-segment degeneration in patients with degenerative lumbar spinal stenosis after posterior instrumented lumbar fusion.

OBJECTIVE: This study aimed to quantify pre- and postoperative paraspinal muscular variation following posterior lumbar interbody fusion (PLIF) in patients with degenerative lumbar spinal stenosis (DLSS) and measure the association of this variation with adjacent-segment degeneration (ASD). METHODS: Data from 149 patients who underwent L4-S1 PLIF for DLSS were collected. Patients were divided into radiologically confirmed ASD and control groups according to follow-up radiological findings. MRI was performed before surgery and at the last follow-up. Muscular parameters including the relative cross-sectional area (rCSA), relative functional cross-sectional area (rFCSA), relative total cross-sectional area (rTCSA), and fatty infiltration (FI) of the multifidus (MF), erector spinae (ES), and psoas major (PM) muscles were measured on preoperative and follow-up L2-S1 MR images. Logistic regression was used to investigate risk factors for ASD. RESULTS: The rate of radiological ASD was 42.3% at the final follow-up (mean 25.71 ± 8.35 months). At surgical levels, the rFCSA and rTCSA of the MF and ES muscles decreased. The FI of the MF from L2-3 to L5-S1 and ES muscles at L5-S1 significantly increased after surgery, while the rFCSA and rTCSA of the PM muscle increased and its FI decreased. At adjacent levels, the rFCSA and rTCSA of the MF muscle and rTCSA of the ES muscle decreased and the FI of the MF muscle increased postoperatively (p < 0.05), but the rFCSA and rTCSA of the PM muscle increased and its FI decreased (p < 0.05). The FIs of the MF, ES, and PM muscles at adjacent levels significantly differed between the ASD and control groups. Logistic regression analysis indicated that higher BMI (p = 0.002) and FI of the PM muscle at adjacent levels (p = 0.025) were significant risk factors for ASD. CONCLUSIONS: The functional area decreased in the MF and ES muscles and increased in the PM muscle after L4-S1 PLIF. A compensatory postoperative decrease in FI of the PM muscle at the adjacent level was a protective factor for ASD in DLSS patients after PLIF.

Preoperative Prediction of Perineural Invasion in Oesophageal Squamous Cell Carcinoma Based on CT Radiomics Nomogram: A Multicenter Study.

RATIONALE AND OBJECTIVES: To investigate the value of computed tomography (CT) radiomics nomogram in the preoperative prediction of perineural invasion (PNI) in oesophageal squamous cell carcinoma (ESCC) through a multicenter study. MATERIALS AND METHODS: We retrospectively collected postoperative pathological data of 360 ESCC patients with definite PNI status (131 PNI-positive and 229 PNI-negative) from two centres. Radiomic features were extracted from the arterial-phase CT images, and the least absolute shrinkage and selection operator and logistic regression algorithm were used to screen valuable features for identifying the PNI status and calculating the radiomics score (Rad-score). A radiomics nomogram was established by integrating the Rad-score and clinical risk factors. A receiver operating characteristic curve was used to evaluate model performance, and decision curve analysis was used to evaluate the predictive performance of the radiomics nomogram in the training, internal validation, and external validation sets. RESULTS: Twenty radiomics features were extracted from a full-volume tumour region of interest to construct the model, and the radiomics nomogram combined with radiomics features and clinical risk factors was superior to the clinical and radiomics models in predicting the PNI status of ESCC patients. The area under the curve values of the radiomics nomogram in the training, internal validation, and external validation sets were 0.856 (0.794-0.918), 0.832 (0.742-0.922), and 0.803 (0.709-0.898), respectively. CONCLUSION: The radiomics nomogram based on CT has excellent predictive ability; it can non-invasively predict the preoperative PNI status of ESCC patients and provide a basis for preoperative decision-making.

Detecting Muscle Invasion of Bladder Cancer: An Application of Diffusion Kurtosis Imaging Ratio and Vesical Imaging-Reporting and Data System.

BACKGROUND: Independent factors are needed to supplement vesical imaging-reporting and data system (VI-RADS) to improve its ability to identify muscle invasive bladder cancer (MIBC). PURPOSE: To assess the correlation between MIBC and diffusion kurtosis imaging (DKI) ratio, VI-RADS, and other factors (such as tumor location). STUDY TYPE: Retrospective. POPULATION: Sixty-eight patients (50 males and 18 females; age: 70.1 ± 9.5 years) with bladder urothelial carcinoma. FIELD STRENGTH/SEQUENCE: 1.5 T, conventional diffusion-weighted imaging (DWI), and DKI (single shot echo-planar sequence). ASSESSMENT: Three radiologists independently measured the diffusion parameters of each bladder cancer (BCa) and obturator internus, including the mean apparent diffusion coefficient (ADCmean), mean kurtosis (MK), and mean diffusion (MD). And the ratio of diffusion parameters between BCa and obturator internus was calculated (diffusion parameter ratio = bladder cancer:obturator internus). Based on the VI-RADS, the target lesions were independently scored. Furthermore, the actual tumor-wall contact length (ACTCL) and absolute tumor-wall contact length (ABTCL) were measured. STATISTICAL TESTS: Multicollinearity among independent variables was evaluated using the variance inflation factor (VIF). Multivariable logistic regression analysis was used to determine the independent risk factors of MIBC. The receiver operating characteristic curve was used to evaluate the efficacy of each variable in detecting MIBC. The DeLong test was used to compare the area under the curve (AUC). A P < 0.05 was considered statistically significant. RESULTS: MKratio (median: 0.62) and VI-RADS were independent risk factors for MIBC. AUCs for MKratio, VI-RADS, and MKratio combined with VI-RADS in assessing MIBC were 0.895, 0.871, and 0.973, respectively. MKratio combined with VI-RADS was more effective in diagnosing MIBC than VI-RADS alone. DATA CONCLUSIONS: MKratio has potential to assist the assessment of MIBC. MKratio can be used as a supplement to VI-RADS for detecting MIBC. LEVEL OF EVIDENCE: 4 TECHNICAL EFFICACY: Stage 2.

Neurosurgical robots in China: State of the art and future prospect.

Neurosurgical robots have developed for decades and can effectively assist surgeons to carry out a variety of surgical operations, such as biopsy, stereo-electroencephalography (SEEG), deep brain stimulation (DBS), and so forth. In recent years, neurosurgical robots in China have developed rapidly. This article will focus on several key skills in neurosurgical robots, such as medical imaging systems, automatic manipulator, lesion localization techniques, multimodal image fusion technology, registration method, and vascular imaging technology; introduce the clinical application of neurosurgical robots in China, and look forward to the potential improvement points in the future based on our experience and research in the field.

Discovery of 4-((3,4-dichlorophenyl)amino)-2-methylquinolin-6-ol derivatives as EGFR and HDAC dual inhibitors.

In patients with non-small cell lung cancer (NSCLC), the standard therapy consists of selective tyrosine kinase inhibitors that target epidermal growth factor receptors (EGFR). Nonetheless, their clinical utility is primarily limited by the development of resistance to drugs. HDAC inhibitors have been shown in studies to reduce the level of EGFR that is expressed and downregulate the EGFR-induced phosphorylation of AKT and ERK. Therefore, dual inhibitors of EGFR and HDAC provide a potential approach as combination treatment synergistically inhibited the growth of NSCLC. Herein, we examined the EGFR inhibition effect of twenty compounds which designed and synthesized by us previously. Among them, compounds 12c and 12d exhibited powerful antiproliferative activity against the NCI-H1975 cell line with IC50 values of 0.48 ± 0.07 and 0.35 ± 0.02 µM, correspondingly. In cell-free kinase assays, both 12c and 12d demonstrated target-specific EGFR inhibition against wild type (EGFRwt). Furthermore, the expression of EGFR and phosphorylation of the EGF-induced pathways were significantly suppressed under the treatment of 12c and 12d. Besides, both histones H3 and H4 exhibited increased levels of acetylation following 12c and 12d treatment. The animal experiments shown that 12d could prevent the growth of tumor, inhibited the expression of EGFR and the phosphorylation levels of p70 S6K, AKT and p38 MAPK in vivo, and did not cause organ damage to the mice during the experiment. Overall, the results illustrated that compound 12c and 12d could serve as effective EGFR and HDAC dual inhibitors in NSCLC cells. Our work offers an alternative strategy for NSCLC therapy.

Site-Specific Modification of Single Domain Antibodies by Enzyme-Immobilized Magnetic Beads.

Nanobodies as imaging agents and drug conjugates have shown great potential for cancer diagnostics and therapeutics. However, site-specific modification of a nanobody with microbial transglutaminase (mTGase) encounters problems in protein separation and purification. Here, we describe a facile yet reliable strategy of immobilizing mTGase onto magnetic beads for site-specific nanobody modification. The mTGase immobilized on magnetic beads (MB-mTGase) exhibits catalytic activity nearly equivalent to that of the free mTGase, with good reusability and universality. Magnetic separation simplifies the protein purification step and reduces the loss of nanobody bioconjugates more effectively than size exclusion chromatography. Using MB-mTGase, we demonstrate site-specific conjugation of nanobodies with fluorescent dyes and polyethylene glycol molecules, enabling targeted immunofluorescence imaging and improved circulation dynamics and tumor accumulation in vivo. The combined advantages of MB-mTGase method, including high conjugation efficiency, quick purification, less protein loss, and recycling use, are promising for site-specific nanobody functionalization and biomedical applications.

Role of the Paraspinal Muscles in the Sagittal Imbalance Cascade: The Effects of Their Endurance and of Their Morphology on Sagittal Spinopelvic Alignment.

BACKGROUND: The role of paraspinal muscle degeneration in the cascade of sagittal imbalance is still unclear. This study aimed to compare paraspinal muscle degeneration in the 4 stages of sagittal imbalance: sagittal balance (SB), compensated sagittal balance (CSB), decompensated sagittal imbalance (DSI), and sagittal imbalance with failure of pelvic compensation (SI-FPC). In addition, it aimed to compare the effects paraspinal muscle endurance and morphology on sagittal spinopelvic alignment in patients with lumbar spinal stenosis. METHODS: A cross-sectional study of 219 patients hospitalized with lumbar spinal stenosis was performed. The isometric paraspinal extensor endurance test and evaluation of atrophy and fat infiltration of the paraspinal extensor muscles and psoas major on magnetic resonance imaging were performed at baseline. Spinopelvic parameters including lumbar lordosis, pelvic tilt, sacral slope, pelvic incidence, and the sagittal vertical axis were measured. RESULTS: The patients with lumbar spinal stenosis were divided into 67 with SB, 85 with CSB, 49 with DSI, and 17 with SI-FPC. There were significant differences in paraspinal muscle endurance and morphology among the 4 groups. Furthermore, the SI-FPC group had poorer paraspinal muscle endurance than either the SB or the CSB group. In multiple linear regression analysis, paraspinal muscle endurance and the relative functional cross-sectional area of the paraspinal extensor muscles were the independent predictors of the sagittal vertical axis, and the relative functional cross-sectional area of the psoas major was the independent predictor of relative pelvic version. CONCLUSIONS: This study indicated that paraspinal muscle degeneration is not only an initiating factor in pelvic retroversion but also a risk factor for progression from a compensated to a decompensated stage. Specifically, the impairment of muscle endurance in the CSB stage may be the reason why patients experience failure of pelvic compensation. In addition, paraspinal muscle endurance and muscle morphology (relative functional cross-sectional area of the paraspinal extensor muscles and psoas major) had different clinical consequences. LEVEL OF EVIDENCE: Prognostic Level II . See Instructions for Authors for a complete description of levels of evidence.

The Role of Thoracic Tilt in Identifying Thoracic Compensation and Predicting Proximal Junctional Kyphosis in Degenerative Lumbar Scoliosis.

STUDY DESIGN: A retrospective cohort study of consecutive patients. OBJECTIVE: To investigate the clinical value of thoracic tilt (TT) in characterizing thoracic compensation and predicting proximal junctional kyphosis (PJK) in degenerative lumbar scoliosis (DLS). SUMMARY OF BACKGROUND DATA: Thoracic compensation has been shown to be associated with the development of PJK, while thoracic shape and morphology in patients with DLS remain understudied. METHODS: Patients with DLS who underwent long-segment fusion were divided into a PJK group and a non-PJK group. Asymptomatic elderly volunteers were recruited as healthy controls. Thoracic parameters were measured in both cohorts, including the TT, T1-L1 pelvic angle (TLPA), T12 slope, thoracic kyphosis (TK, T4-T12), global thoracic kyphosis (GTK, T1-T12), and thoracolumbar kyphosis (TLK, T10-L2). Multivariate logistic regression was used to assess the association between TT and the development of PJK, adjusting for confounders. Multivariate linear regression was used to establish the predictive formula for TT. RESULTS: A total of 126 patients with DLS were enrolled, of which 37 (29.4%) developed PJK. Compared with 110 healthy controls, DLS patients had significantly greater TT, TLPA, T12 slope, and TLK as well as smaller TK and GTK (all P <0.001). Preoperatively, the PJK group showed significantly greater TT ( P =0.013), TLPA ( P <0.001), and TLK ( P =0.034) than the non-PJK group. No significant differences were found in TK and GTK before surgery. Postoperatively, the PJK group showed significantly greater TT ( P <0.001), TLPA ( P <0.001), TLK ( P <0.001), and proximal junctional angle ( P <0.001). Multivariate logistic regression analysis showed that greater postoperative TT was associated with the development of PJK. Multivariate linear regression analysis suggested that the regression formula was postoperative TT=0.675×T12slope+0.412×TK+0.158×TLK-4.808 ( R2 =0.643, P <0.001). CONCLUSIONS: The novel sagittal parameter TT can be used for the evaluation of thoracic compensation. Greater preoperative TT might represent a decompensated state of TK. Rebalancing the TT in a sagittal neutral position might help to prevent PJK in patients with DLS.

Effects of vitamin D supplementation on ovulation and pregnancy in women with polycystic ovary syndrome: a systematic review and meta-analysis.

Objective: To evaluate the effect of vitamin D supplementation on pregnancy and ovulation in patients with polycystic ovary syndrome. Method: We searched Pubmed, Medline (via Ovid, 1974 to 2020), EMBASE (via Ovid, 1974 to 2020), Cochrane Central Register of Controlled Trials (via Ovid), Web of Science, CNKI, WangFang and the Vip database from inception until April 2021. Two researchers independently screened articles, collected data and evaluated the quality, with Review manager 5.3 for meta-analysis. Results: Totally 20 randomized controlled studies with 1961 subjects were included. Meta analysis showed that pregnancy rate [RR=1.44 (1.28, 1.62), p<0.00,001], ovulation rate [RR=1.42 (1.14, 1.78), p=0.002] and matured oocytes rate [RR=1.08 (1.03, 1.13), p=0.002] of vitamin D supplementation group were significantly higher than those of control group. Meanwhile, early miscarriage rate [RR=0.44 (0.30, 0.66), p<0.00,001], androgen level [MD=-2.31 (-3.51, -1.11), p=0.0002], luteinizing hormone [MD=-1.47 (-2.57, -0.36), p=0.009], follicle stimulating hormone [MD=-0.15 (-0.24, -0.05), p=0.002], and premature delivery rate [RR=0.38, 95% CI (0.21, 0.70), p=0.002] were declined significantly than the controls. However, only one article suggested that the progesterone [MD=6.52 (4.52, 8.52), p<0.05] in the vitamin D intervention group was increased. There was no notable difference in the biochemical pregnancy rate [RR=0.95 (0.55, 1.63), p=0.84], gestational hypertension rate [RR=0.40, 95% CI (0.15, 1.11), p=0.08], gestational diabetes mellitus rate [RR=0.27, 95% CI (0.05, 1.39), p=0.11], fertilization rate [RR=1.05 (1.00, 1.10), p=0.04], cleavage rate [RR=1.03 (0.99, 1.06), p=0.17], high-quality embryo rate [RR=1.08 (0.98, 1.20), p=0.10], endometrial thickness [MD=0.10], 77 (-0.23, 1.77), p=0.13], estrogen level [MD=-0.34 (-1.55, 0.87), p=0.59], LH/FSH [MD=-0.14, 95% CI (-0.48, 0.20), p=1.00] and anti-Mullerian hormone [MD=-0.22 (-0.65, 0.21), p=0.32]. Conclusion: Vitamin D supplementation contribute to the higher pregnancy and ovulation rates, and lower androgen, LH, FSH and early miscarriage rates in women with PCOS, regardless of the use of ovulation induction drugs or assisted reproductive technologies. However, no significant improvement was observed in fertilization rate or cleavage rate. Due to the limitation in quality of involved studies, more high-quality RCTs are needed for further validation. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO, identifier CRD42021250284.

Use and misuse of random forest variable importance metrics in medicine: demonstrations through incident stroke prediction.

BACKGROUND: Machine learning tools such as random forests provide important opportunities for modeling large, complex modern data generated in medicine. Unfortunately, when it comes to understanding why machine learning models are predictive, applied research continues to rely on 'out of bag' (OOB) variable importance metrics (VIMPs) that are known to have considerable shortcomings within the statistics community. After explaining the limitations of OOB VIMPs - including bias towards correlated features and limited interpretability - we describe a modern approach called 'knockoff VIMPs' and explain its advantages. METHODS: We first evaluate current VIMP practices through an in-depth literature review of 50 recent random forest manuscripts. Next, we recommend organized and interpretable strategies for analysis with knockoff VIMPs, including computing them for groups of features and considering multiple model performance metrics. To demonstrate methods, we develop a random forest to predict 5-year incident stroke in the Sleep Heart Health Study and compare results based on OOB and knockoff VIMPs. RESULTS: Nearly all papers in the literature review contained substantial limitations in their use of VIMPs. In our demonstration, using OOB VIMPs for individual variables suggested two highly correlated lung function variables (forced expiratory volume, forced vital capacity) as the best predictors of incident stroke, followed by age and height. Using an organized analytic approach that considered knockoff VIMPs of both groups of features and individual features, the largest contributions to model sensitivity were medications (especially cardiovascular) and measured medical risk factors, while the largest contributions to model specificity were age, diastolic blood pressure, self-reported medical risk factors, polysomnography features, and pack-years of smoking. Thus, we reach very different conclusions about stroke risk factors using OOB VIMPs versus knockoff VIMPs. CONCLUSIONS: The near-ubiquitous reliance on OOB VIMPs may provide misleading results for researchers who use such methods to guide their research. Given the rapid pace of scientific inquiry using machine learning, it is essential to bring modern knockoff VIMPs that are interpretable and unbiased into widespread applied practice to steer researchers using random forest machine learning toward more meaningful results.

Role of the tumor microenvironment in malignant melanoma organoids during the development and metastasis of tumors.

Malignant melanoma (MM) is the most metastatic and aggressive form of skin cancer, and carries a high risk of death. Immune-checkpoint inhibitor therapy and molecular-targeted therapy can prolong the survival of patients with advanced MM significantly. However, the low response rate and inevitable drug resistance prevent further improvements in efficacy, which is closely related to the tumor microenvironment (TME). The TME refers to the tumor stroma, including fibroblasts, keratinocytes, immune cells, soluble molecules, and extracellular matrix (ECM). The dynamic interaction between the TME and tumor cells is very important for the growth, local invasion, and metastatic spread of tumor cells. A patient-derived organoid (PDO) model involves isolation of tumor tissue from patients with MM and culturing it in vitro in a three-dimensional pattern. Compared with traditional cultivation methods, the PDO model preserves the heterogeneity of the tissue structure of MM and demonstrates the interaction between MM cells and the TME. It can reproduce the characteristics of proliferation, migration, and invasion of MM cells, and better simulate the structural function of MM in vivo. This review explores the role of each TME component in development of the PDO model. This review will provide a reference for research on the drug screening and targeted treatment using PDOs, particularly for the immunotherapy of MM.

Kaempferitrin, a major compound from ethanol extract of Chenopodium ambrosioides, exerts antitumour and hepatoprotective effects in the mice model of human liver cancer xenografts.

OBJECTIVES：: This study aimed to investigate antitumour effect and possible toxicity of kaempferitrin, the major compound from ethanol extract of Chenopodium ambrosioides, in the mice model of human liver cancer xenografts. METHODS：: Forty mice bearing SMMC-7721 cells xenografts were divided into control group (not treated) and three groups orally administered with ethanol extract of C. ambrosioides, kaempferol (positive control) and kaempferitrin for 30 days. Antitumour effect was evaluated by measurement of tumour growth, histological examinations of tumours, flow cytometry detection of splenic CD19+ B lymphocytes and CD161+ Natural Killer cells, biochemical measurements of serum levels of tumour necrosis factor-α, interleukin-6, interferon-γ, malonaldehyde, 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azinobis-(3-ethylbenz thiazoline-6-sulphonate) radicals. Toxicity was evaluated by histological examinations of livers and measurements of serum levels of aspartate transaminase, alanine transaminase, total bilirubin, direct bilirubin, malonaldehyde and hepatic malonaldehyde level. KEY FINDINGS: Kaempferitrin significantly (P < 0.05) decreased tumour volume, mass and cell number. Antitumour effect was due to induction of tumour cells necrosis and apoptosis, stimulation of splenic B lymphocytes, decreases of radicals and malonaldehyde. Kaempferitrin did not change liver structure, and decreased serum levels of transaminases, bilirubin, malonaldehyde and hepatic malonaldehyde level. CONCLUSIONS: Kaempferitrin exerts antitumour and hepatoprotective effects.