Exosomes derived from human adipose mesenchymal stem cells ameliorate hepatic fibrosis by inhibiting PI3K/Akt/mTOR pathway and remodeling choline metabolism.

Liver fibrosis is a chronic liver disease with the presence of progressive wound healing response caused by liver injury. Currently, there are no approved therapies for liver fibrosis. Exosomes derived from human adipose mesenchymal stem cells (hADMSCs-Exo) have displayed a prominent therapeutic effect on liver diseases. However, few studies have evaluated therapeutic effect of hADMSCs-Exo in liver fibrosis and cirrhosis, and its precise mechanisms of action remain unclear. Herein, we investigated anti-fibrotic efficacy of hADMSCs-Exo in vitro and in vivo, and identified important metabolic changes and the detailed mechanism through transcriptomic and metabolomic profiling. We found hADMSCs-Exo could inhibit the proliferation of activated hepatic stellate cells through aggravating apoptosis and arresting G1 phase, effectively inhibiting the expression of profibrogenic proteins and epithelial-to-mesenchymal transition (EMT) in vitro. Moreover, it could significantly block collagen deposition and EMT process, improve liver function and reduce liver inflammation in liver cirrhosis mice model. The omics analysis revealed that the key mechanism of hADMSCs-Exo anti-hepatic fibrosis was the inhibition of PI3K/AKT/mTOR signaling pathway and affecting the changes of metabolites in lipid metabolism, and mainly regulating choline metabolism. CHPT1 activated by hADMSCs-Exo facilitated formation and maintenance of vesicular membranes. Thus, our study indicates that hADMSCs-Exo can attenuate hepatic stellate cell activation and suppress the progression of liver fibrosis, which holds the significant potential of hADMSCs-Exo for use as extracellular nanovesicles-based therapeutics in the treatment of liver fibrosis and possibly other intractable chronic liver diseases.

Prognostic value of inflammation-immunity-nutrition score in patients with hepatocellular carcinoma treated with anti-PD-1 therapy.

BACKGROUND: There are no validated biomarkers that can predict the clinical benefit of immune checkpoint blockers against the programmed cell death protein 1 (PD-1) treatments in hepatocellular carcinoma (HCC). This study aimed to investigate the prognostic value of inflammation-immunity-nutrition score (IINS) in patients with HCC treated with anti-PD-1 therapy. METHODS: A consecutive series of 101 HCC patients treated with PD-1 inhibitors in Sichuan Provincial People's Hospital between January 2018 and August 2020 were enrolled in the retrospective study. IINS (0-6) was constructed based on pretreatment high-sensitivity C-reactive protein (hsCRP), lymphocyte (LYM), and albumin (ALB). The patients were divided into high and low IINS groups according to IINS values. Prognostic values of each variable were evaluated with univariate and multivariate time-dependent Cox regression analyses. Survival curves were calculated and compared using the Kaplan-Meier method and log-rank test. The prognostic performance of IINS was further compared with that of other traditional prognostic indicators by receiver operating characteristic (ROC) curve and the areas under the ROC curve. RESULTS: Patients with low IINS had longer overall survival (OS) (HR: 4.711, 95% CI: 1.80-12.37, p = .001) and progression-free survival (HR: 3.411, 95% CI: 1.79-6.51, p < .0001) than those with high IINS. The multivariate analysis identified IINS (HR: 3.746, 95% CI: 1.05-13.38, p = .042) and tumor number (HR: 5.111, 95% CI: 1.075-24.299, p = .04) as independent prognostic factors. According to ROC analysis, IINS (AUC =0.729, 95% CI: 0.597-0.861, p = .002) presented better prognostic performance than other traditional prognostic indicators. The area of the IINS-CA19-9 under the ROC curve (AUC) was higher than that of the IINS or CA19-9 levels for the prediction of OS. CONCLUSION: The results suggest that IINS may be an independent prognostic indicator for HCC patients treated with anti-PD-1 therapy. IINS-CA19-9 classification may be more effective in predicting clinical benefit of anti-PD-1 therapy in HCC patients.

Risk factors for sleep quality disturbances in patients with lumbar spinal stenosis before operation.

PURPOSE: We aimed to explore the risk factors of preoperative sleep quality in patients with lumbar spinal stenosis (LSS) and the association of sleep-related beliefs with sleep quality in these patients. METHODS: Sleep quality and related risk factors of sleep quality disturbances in patients with LSS preoperatively were assessed by questionnaires. Pittsburgh Sleep Quality Index (PSQI) for sleep quality, Oswestry Disability Index (ODI) for clinical outcomes, Visual Analog Scale for Pain (VAS Pain), Self-Rating Anxiety Scale (SAS) for anxiety level, and Dysfunctional Beliefs and Attitudes about Sleep (DBAS-16) for sleep-related beliefs were assessed. Bivariate logistic regression analysis was used to assess the risk factors of sleep quality disturbances. RESULTS: A total of 227 patients were enrolled, mean age 64 years (SD 13.1), 119 women (52%). The incidence of sleep quality disturbances in patients was 37% (83/227). Increased DBAS-16 scores (OR = 0.781; 95% CI, 0.725-0.841; p < 0.001) significantly decreased the probability of developing sleep quality disturbances, while increased anxiety levels (OR = 1.241; 95% CI, 1.152-1.337; p < 0.001) significantly increased the probability of developing sleep quality disturbances in patients. Factors including educational level, increased age, sex, preoperative length of stay, VAS Pain scores, and ODI scores showed no significant association and were therefore excluded from the model. CONCLUSIONS: High levels of anxiety and mistaken sleep-related beliefs were risk factors of sleep quality disturbances in patients with LSS before surgery. The more mistaken sleep-related beliefs were, the greater the probability of sleep disturbances.

Preliminary experience in laparoscopic resection of hepatic hydatidectocyst with the Da Vinci Surgical System (DVSS): a case report.

BACKGROUND: At present, Da Vinci robotic assisted hepatectomy has been routinely carried out in conditional units. But there is no report concerning the use of Da Vinci robots for hepatic hydatid cystectomy and experience on this aspect is seldom mentioned before. This study was to summarize the preliminary experience in laparoscopic resection of hepatic hydatidectocyst with the Da Vinci Surgical System (DVSS). CASE PRESENTATION: A 29-year-old female diagnosed as hepatic hydatid in the right anterior lobe of liver was treated with laparoscopic resection by the DVSS under general anesthesia. Appropriate disposal of tumor cell in vascular system and disinfection of surgical field with hypertonic saline were conducted. The hepatic hydatidectocyst was resected completely with an operation time of 130 min, an intraoperative blood loss of 200 ml and a length of hospital stay for five days. The vital signs of patient were stable and no cyst fluid allergy occurred after operation. CONCLUSIONS: Our result showed that laparoscopic resection of hepatic hydatidectocyst by using the DVSS is safe and feasible on the basis of hospitals have rich experience in treatment of cystic echinococcosisliver, resection with DVSS and laparoscopic excision.

Intraoperative blood loss, postoperative drainage, and recovery in patients undergoing lumbar spinal surgery.

BACKGROUND: Spine surgery is widely accepted as an effective management for patients with lumbar disc herniation; however, the factors influencing intraoperative procedure and prognosis are not fully understood. The present study was aimed to identify the factors influencing intraoperative blood loss, postoperative drainage volume, and recovery in patients undergoing spinal surgery. METHODS: We retrospectively analyzed the clinical data of 183 consecutive patients with lumbar disc herniation who underwent spine surgery. The clinical characteristics, operation procedure, and outcome were documented and the correlations were analyzed. RESULTS: There were significant differences between one-level and two-level operations in the bleeding volumes of male (P = 0.005) and female (P = 0.002) patients, and in final drainage of male (P = 0.043) and female (P = 0.003) patients. The blood loss was correlated with the operation duration. There were differences in intraoperative bleeding and final drainage between groups with one-level and two-level operations. Additionally, there were differences in intraoperative autologous blood transfusion among various groups. There were significant differences in intraoperative bleeding between autologous blood transfusion and non-transfusion groups. CONCLUSIONS: The key factors affecting the intraoperative blood loss and postoperative drainage volume include operation methods, operation duration, blood-transfusion modes, and usage of anticoagulants. These results should be taken into consideration in the attempt to optimize operation procedure and improve post-operative recovery.

High-intensity focused ultrasound ablation induced apoptosis in human hepatocellular carcinoma.

BACKGROUND/AIMS: To evaluate the effect of high-intensity ultrasound (HIFU) ablation on human hepatocellular carcinoma tissues and apoptotic proteins (bcl-2 and p-53). METHODOLOGY: Patients with hepatocellular carcinoma at stage B were treated with HIFU ablation. Levels of bcl-2 and p53 protein and the apoptosis rate were evaluated both in the pre-treatment and post-treatment tissue specimens using immunochemistry and TUNEL methods, respectively. RESULTS: After HIFU ablation, p53 protein levels were significantly increased around the coagulation necrosis area, whereas, the level of bcl-2 was significantly decreased. More apoptosis cells were found post ablation compared with those in the pretreatment tissues. Additionally, no significant correlation was found between p53/bcl-2 levels and apoptotic index. CONCLUSIONS: HIFU ablation may exert promote the apoptosis of hepatocellular carcinoma cells and the effect has a closely association with the change of p53 and bcl-2 expression.

Clinical outcome of autologous hematopoietic stem cell infusion via hepatic artery or portal vein in patients with end-stage liver diseases.

OBJECTIVE: To investigate the efficacy of hematopoietic stem cell (HSC) transplantation via the hepatic artery vs. the portal vein for end-stage liver disease (ESLD). METHODS: Patients with hepatic decompensation were prospectively recruited from September 2010 to September 2012 to receive HSC transplantation via the hepatic artery or the portal vein. Liver function was examined at 3, 6, and 12 months after transplantation. Liver biopsy Results were analyzed using the Knodell score. RESULTS: Eighty patients (58 males and 22 females) were enrolled in the study. The Child-Pugh score was grade B in 69 cases, and grade C in the remaining 11 cases. HSC transplantation was performed via the portal vein in 36 patients and via the hepatic artery in 44 patients. ALT levels decreased while serum albumin levels increased significantly in both groups at 6 and 12 months after HSC transplantation (P<0.05 compared with pre-transplantation levels). Total bilirubin levels decreased significantly in both groups at 3, 6, and 12 months after HSC transplantation (P<0.05 compared with pre-transplantation levels). Additionally, prothrombin time decreased in both groups at 12 months after HSC transplantation (P<0.05 compared with pre-transplantation level). There were no significant differences in ALT, total bilirubin and prothrombin time between the two groups either before or after transplantation. Moreover, Knodell score decreased significantly at 6 and 12 months. Histological examination showed that liver cell edema, degeneration, necrosis, and inflammation were significantly relieved at 3, 6, and 12 months after transplantation. The incidence of portal vein thrombosis, upper gastrointestinal bleeding, and hepatic encephalopathy were 1.25%, 3.75%, and 2.5% respectively. The one-year survival rate was 100%. CONCLUSIONS: Autologous HSC transplantation improves liver function and histology in ESLD patients. The administration route of HSC has no significant impact on the efficacy of transplantation.

[Imaging assessing symptomatic vertebral compression fracture to be treated by vertebroplasty and kyphoplasty in osteoporosis patients].

OBJECTIVE: To evaluate the values of radiology, computed tomography (CT) and magnetic resonance imaging (MRI) in assessing symptomatic osteoporosis vertebral compression fractures in vertebroplasty and kyphoplasty. METHODS: Retrospective reviews were conducted for 41 osteoporotic vertebral compression fractures patients with preoperative CT (with 3D reconstruction)/MRI (with fat saturation sequence) and preoperative and postoperative radiological imaging data. All patients underwent percutaneous vertebroplasty (PVP) or balloon kyphoplasty (BKP) when conservative treatment failed to offer pain relief. RESULTS: CT scan showed 52 vertebral fractures while MRI detected a total of 70 lesions of vertebral body, including 44 new fractures, 8 old fractures and 18 bone bruises. Eight old fractures and 2 mild bone bruises were treated conservatively. In conjunctions with MRI, especially fat saturation sequence, 60 therapeutic vertebral bodies were visualized. It was superior to CT. As demonstrated by preoperative and postoperative radiology, vertebral body height improved and Cobb Angle slightly decreased for 60 lesions. CONCLUSION: For vertebral compression fractures in osteoporosis patients, preoperative CT and MRI examinations have important clinical significance in providing treatment guidance and assessing postoperative outcomes.

[Effects of different transfusion on intraoperative blood loss and postoperative drainage volume in posterior lumbar spinal surgery].

OBJECTIVE: To explore the effects of autotransfusion on intraoperative blood loss and postoperative drainage volume in posterior lumbar spinal surgery. METHODS: From June 2010 to January 2012, a total of 183 patients with lumbar disease were enrolled. There were 74 males and 109 females with an average age of 56.6 (31-81) years. There were lumbar disc herniation (n = 80), lumbar stenosis (n = 69), lumbar spondylolisthesis (n = 32) and discogenic pain (n = 2). And one-level (n = 83) and two-level (n = 100) operations were performed. The treatments were non-transfusion (n = 34), auto-blood (n = 109) and allogeneic blood (n = 20) and both (n = 20).Operative duration, operative level, intraoperative autologous blood transfusion, intraoperative allogeneic blood transfusion, intraoperative blood loss, immediate drainage volume, duration of drainage tube and final drainage volume were recorded. RESULTS: The mean intraoperative blood loss was (369.4 ± 284.0) ml and (568.8 ± 398.7) ml for two-level operation, there were significant differences between them(t = 3.94, P < 0.001). The final drainage volume was (186.3 ± 135.6) ml for one-level patients and (301.4 ± 286.8) ml for two-level patients, there were significant differences between them (t = 3.56, P < 0.05). The total intraoperative blood loss was (478.3 ± 364.3) ml and total final drainage volume was (249.2 ± 237.4) ml, there were significant differences between them(P < 0.001). There were significant differences in blood loss between autologous blood transfusion for non-transfusion groups (P < 0.001) .However, there was no difference for immediate drainage and final drainage comparing with other blood transfusion methods (P > 0.05) . CONCLUSION: Using autologous blood transfusion alone during operation has significant difference for inoperative blood loss compared with non-transfusion. Yet there is no effect on immediate or final drainage volume.

[Autologous peripheral blood CD34+ stem cells transplanted into 100 patients with advanced cirrhosis].

OBJECTIVE: To investigate whether transplantation of autologous peripheral blood CD34+ stem cells is a viable approach for treating patients with advanced cirrhosis,which is currently hindered by a shortage in liver donors. METHODS: A total of 100 patients with advanced cirrhosis and who had failed to respond to conservative therapy were recruited for transplantation of autologous peripheral blood CD34+ stem cells.The success of transplantation was investigated 6-and 12-months later by measuring markers of liver biosynthesis function (coagulation,albumin level,indocyanine green clearance,Child-Pugh score) and assessing pathological changes (Knodell score) and morphologic changes in the liver tissue.Complications were also recorded during follow-up. RESULTS: The 1-year cumulative survival rate was 100%. Fifty-two patients with massive ascites showed gradual reduction and disappearance of the ascites.Four patients experienced upper gastrointestinal bleeding and three patients developed with hepatic encephalopathy (I-II degree) at 3 months post-transplantation.All patients showed significantly improved liver biosynthesis function,liver elasticity and Knodell score after transplantation (P less than 0.05). CONCLUSION: Autologous peripheral blood CD34+ stem cell transplantation is a safe and effective treatment for advanced cirrhosis,and has high cost-benefit since it improves liver function,liver histology,and quality of life.

Diffusion-induced lithium isotopic heterogeneity in olivines from peridotites of an oceanized mantle lithosphere at the Yunzhug ophiolite (central Tibet).

This paper reports lithium concentrations and isotopic compositions of olivines in the oceanized subcontinental lithospheric mantle (SCLM) peridotites of the Tibetan Yunzhug ophiolite. The results show systematic Li isotope changes with distance from the rim of olivine grains. δ7Li values of olivine in dunites decrease from + 10.46 to + 1.33‰ with increasing distance to olivine rim from 26.15 to 124.71 µm. A negative correlation of δ7Li and Li content in olivine from dunite and harzburgite indicates recent diffusive ingress of Li into the peridotites. The extremely heavy Li isotopic composition requires the seawater or seawater alteration endmember in the mixing model, and reveals Li diffusion from seawater into olivine. As in dunites, olivines in a harzburgite sample show similar variations in δ7Li as a function of distance from the grain rim (e.g., 6.01 to 1.73 in sample 14YZ13). We suggest that the behavior of Li in the oceanized SCLM peridotites may be controlled by Li diffusion from seawater, as Li activity in the liquid state is higher than the solid state in transporting Li through the olivines in the peridotites. This study supports that seawater Li diffusion is one of the important factors for the heterogeneity of mantle Li isotopes in ophiolites.

hASCs-derived exosomal miR-155-5p targeting TGFßR2 promotes autophagy and reduces pyroptosis to alleviate intervertebral disc degeneration.

Background: Intervertebral disc degeneration (IDD) is a complex chronic disease involving nucleus pulposus cells (NPCs) senescence, apoptosis, autophagy and extracellular matrix (ECM) degradation. In this study, we aimed to investigate the role of human adipose tissue stem cells (hASCs)-derived exosomal miR-155-5p targeting TGFßR2 in IDD and the mechanisms involved. Then miRNA sequencing was performed, and hASCs-derived Exo (hASCs-Exo) was extracted and characterized. Methods: First, NPCs were treated with different concentrations of LPS. Then miRNA sequencing was performed, and hASCs-Exo was extracted and characterized. NPCs were treated with PBS or autophagy inhibitor 3-MA. NPCs were transfected with miR-155-5p mimic, si-TGFßR2 and negative control. Cell viability, apoptosis, ROS, caspase-1+PI, pyroptosis markers, inflammatory cytokines, autophagy markers, Aggrecan, MMP13, and Akt/mTOR pathway-related factors were measured. Bioinformatics prediction and dual-luciferase were performed to verify the binding sites of miR-155-5p to TGFßR2. Finally, we validated the role of hASCs-derived exosomal miR-155-5p on IDD in vivo. Results: LPS promoted pyroptosis of NPCs, and inhibited autophagy and ECM synthesis. MiR-155-5p was characterized as an inflammation-related miRNA in NPCs. HASCs-derived exosomal miR-155-5p inhibited pyroptosis of NPCs and promoted autophagy and ECM synthesis. After bioinformatics prediction and verification, it was found that miR-155-5p targeted TGFßR2. Moreover, miR-155-5p targeted TGFßR2 to promote autophagy and inhibit pyroptosis in NPCs. In vivo experiments revealed that hASCs-derived exosomal miR-155-5p alleviated IDD in rats. Conclusions: HASCs-derived exosomal miR-155-5p alleviated IDD by targeting TGFßR2 to promote autophagy and reduce pyroptosis. Our study may provide a new therapeutic target for IDD. Translational Potential of this Article: HASCs-derived exosomal miR-155-5p is expected to be a biomarker for clinical treatment of IDD. Our study may provide a new therapeutic target for IDD.

A strong short-duration convection near Poyang Lake in daytime of warm season.

Poyang Lake (PL), the biggest freshwater lake in China, is situated in the East Asian Monsoon region, and has an important impact on local convection. In general, PL can result in convection in local region when it is a heat source in nighttime of warm season. However, at around noon on 4 May 2020 (PL is a cold source), a convection was triggered about 20 km west of PL, and rapidly enhanced and resulted in lightning when approaching PL, and then quickly weakened and disappeared after entering the main body of PL. In order to explore the convection formation, several observational data and the Weather Research and Forecasting model were applied in this study. Results show that when the convection approaches PL, its rapid enhancement is induced by PL, and after entering the main body of PL, its quick weakening is also resulted from PL. However, the initiation of the convection is mainly induced by the local topography west of PL under a favorable large-scale background. Mechanism analysis indicates that the strong low-level convergence near the west shore of PL associated with lake-land breeze is responsible for the rapid enhancement of the convection, and the low-level divergence over the main body of PL associated with the lake-land breeze and the increase of the low-level stability induced by cooling of PL jointly result in the quick weakening of the convection. The prevailing southerly wind in low level passes through the local topography (Meiling Mountain) west of PL, and is divided into southwesterly wind (flow around Meiling) and southerly wind (flow over Meiling), and they converge in the north of Meiling, triggering the convection. This study is not only important to deepen the understanding of PL affecting regional weather, but also helpful for improving the refined forecasting of convection near PL.

ST8SIA6-AS1 contributes to hepatocellular carcinoma progression by targeting miR-142-3p/HMGA1 axis.

Hepatocellular carcinoma (LIHC) accounts for 90% of all liver cancers and is a serious health concern worldwide. Long noncoding RNAs (lncRNAs) have been observed to sponge microRNAs (miRNAs) and participate in the biological processes of LIHC. This study aimed to evaluate the role of the ST8SIA6-AS1-miR-142-3p-HMGA1 axis in regulating LIHC progression. RT-qPCR and western blotting were performed to determine the levels of ST8SIA6-AS1, miR-142-3p, and HMGA1 in LIHC. The relationship between ST8SIA6-AS1, miR-142-3p, and HMGA1 was assessed using luciferase assay. The role of the ST8SIA6-AS1-miR-142-3p-HMGA1 axis was evaluated in vitro using LIHC cells. Expression of ST8SIA6-AS1 and HMGA1 was significantly upregulated, whereas that of miR-142-3p was markedly lowered in LIHC specimens and cells. ST8SIA6-AS1 accelerated cell growth, invasion, and migration and suppressed apoptosis in LIHC. Notably, ST8SIA6-AS1 inhibited HMGA1 expression by sponging miR-142-3p in LIHC cells. In conclusion, sponging of miR-142-3p by ST8SIA6-AS1 accelerated the growth of cells while preventing cell apoptosis in LIHC cells, and the inhibitory effect of miR-142-3p was abrogated by elevating HMGA1 expression. The ST8SIA6-AS1-miR-142-3p-HMGA1 axis represents a potential target for the treatment of patients with LIHC.

Artificial intelligence differentiates abdominal Henoch-Schönlein purpura from acute appendicitis in children.

OBJECTIVE: This study aims to construct an artificial intelligence (AI) model capable of effectively discriminating between abdominal Henoch-Schönlein purpura (AHSP) and acute appendicitis (AA) in pediatric patients. METHODS: A total of 6965 participants, comprising 2201 individuals with AHSP and 4764 patients with AA, were enrolled in the study. Additionally, 53 laboratory indicators were taken into consideration. Five distinct artificial intelligence (AI) models were developed employing machine learning algorithms, namely XGBoost, AdaBoost, Gaussian Naïve Bayes (GNB), MLPClassifier (MLP), and support vector machine (SVM). The performance of these prediction models was assessed through receiver operating characteristic (ROC) curve analysis, calibration curve assessment, and decision curve analysis (DCA). RESULTS: We identified 32 discriminative indicators (p < .05) between AHSP and AA. Five indicators, namely the lymphocyte ratio (LYMPH ratio), eosinophil ratio (EO ratio), eosinophil count (EO count), neutrophil ratio (NEUT ratio), and C-reactive protein (CRP), exhibited strong performance in distinguishing AHSP from AA (AUC ≥ 0.80). Among the various prediction models, the XGBoost model displayed superior performance evidenced by the highest AUC (XGBoost = 0.895, other models < 0.89), accuracy (XGBoost = 0.824, other models < 0.81), and Kappa value (XGBoost = 0.621, other models < 0.60) in the validation set. After optimization, the XGBoost model demonstrated remarkable diagnostic performance for AHSP and AA (AUC > 0.95). Both the calibration curve and decision curve analysis suggested the promising clinical utility and net benefits of the XGBoost model. CONCLUSION: The AI-based machine learning model exhibits high prediction accuracy and can differentiate AHSP and AA from a data-driven perspective.

Repair of fingertip defect with reverse digital artery island flap and repair of donor site with digital dorsal advancement flap.

Objective: The reverse digital artery island flap (RDAF) is widely used in repairing fingertip skin defects based on its good appearance and practicability. However, the donor area of the flap needs skin grafting, which can lead to complications. This retrospective study explored the clinical application of digital dorsal advance flap (DDAF) in repairing the donor site of the reverse digital artery island flap. Method: From June 2019 to February 2022, 17 patients with a soft tissue defect of the finger had been restored with the reverse digital artery island flap, and at the same time, the donor area was repaired with digital dorsal advance flap (DDAF). The sensitivity, the active range of motion (ROM) and patient satisfaction were assessed after the operation. Results: All flaps survived completely without skin grafting with only one linear scar. The sensory and motor functions of all patients recovered well. Assessment based on the Michigan Hand Outcomes Questionnaire (MHQ) showed satisfactory functional recovery for all patients. Conclusions: Reconstruction using RDAF combined with DDAF represents an effective alternative for repairing fingertip skin defects.

Cholesterol level influences opioid signaling in cell models and analgesia in mice and humans.

Cholesterol regulates the signaling of µ-opioid receptor in cell models, but it has not been demonstrated in mice or humans. Whether cholesterol regulates the signaling by mechanisms other than supporting the entirety of lipid raft microdomains is still unknown. By modulating cholesterol-enriched lipid raft microdomains and/or total cellular cholesterol contents in human embryonic kidney cells stably expressing µ-opioid receptor, we concluded that cholesterol stabilized opioid signaling both by supporting the lipid raft's entirety and by facilitating G protein coupling. Similar phenomena were observed in the primary rat hippocampal neurons. In addition, reducing the brain cholesterol level with simvastatin impaired the analgesic effect of opioids in mice, whereas the opioid analgesic effect was enhanced in mice fed a high-cholesterol diet. Furthermore, when the records of patients were analyzed, an inverse correlation between cholesterol levels and fentanyl doses used for anesthesia was identified, which suggested the mechanisms above could also be applicable to humans. Our results identified the interaction between opioids and cholesterol, which should be considered in clinics as a probable route for drug-drug interaction. Our studies also suggested that a low cholesterol level could lead to clinical issues, such as the observed impairment in opioid functions.

The prognostic significance of inflammation-immunity-nutrition score on postoperative survival and recurrence in hepatocellular carcinoma patients.

Background: Inflammation, immunity, and nutrition status play important roles in tumorigenesis, progression, and metastasis. This study aimed to evaluate the prognostic value of Inflammation-Immunity-Nutrition Score (IINS) for overall survival (OS) and progression-free survival (PFS) in patients with hepatocellular carcinoma (HCC) undergoing radical surgery. Methods: A total of 204 HCC patients who met the criteria were included in this retrospective study: 144 in the prediction model and 60 in the validation model. IINS was constructed based on the sum of classification scores of preoperative high-sensitivity C-reactive protein (hsCRP), lymphocyte (LYM), and albumin (ALB). The associations between the IINS group and the clinicopathologic characteristics were analyzed using Pearson's χ2 test or Fisher's exact test. Multivariate Cox regression analysis was used to evaluate variables significant on univariate analysis. Kaplan-Meier survival curves were conducted to investigate the prognostic values of IINS, Alpha-fetoprotein (AFP) and IINS-AFP classification. The prognostic performances of all the potential prognostic factors were further compared by receiver operating characteristic (ROC) curve, and time-dependent ROC curve. The internal validation and external validation were used to ensure the credibility of this prediction model. Results: The patients were divided into low and high IINS groups according to the median of IINS. According to multivariate Cox regression analyses, the Barcelona Clinic Liver Cancer (BCLC) Stage (P=0.003), AFP (P=0.013), and IINS (P=0.028) were independent prognostic factors for OS, and BCLC Stage (P=0.009), microvascular invasion (P=0.030), and IINS (P=0.031) were independent prognostic factors for PFS. High IINS group were associated with significantly worse OS and PFS compared with low IINS group (P<0.001; P=0.004). In terms of clinical prognosis, IINS-AFP classification was good in group I, moderate in group II, and poor in group III. Group I had a longer OS (P<0.001) and PFS (P=0.008) compared with group II and III. ROC analysis revealed that IINS-AFP classification had a better prognostic performance for OS (AUC: 0.767) and PFS (AUC: 0.641) than other predictors, excluding its slightly lower predictive power for PFS than IINS. The time-dependent ROC curves also showed that both IINS (12-month AUC: 0.650; 24-month AUC: 0.670; 36-month AUC: 0.880) and IINS-AFP classification (12-month AUC: 0.720; 24-month AUC: 0.760; 36-month AUC: 0.970) performed well in predicting OS for HCC patients. Furthermore, the internal validation and external validation proved that IINS had good predictive performance, strong internal validity and external applicability, and could be used to establish the prediction model. Conclusion: Inflammation-immunity-nutrition score could be a powerful clinical prognostic indicator in HCC patients undergoing radical surgery. Furthermore, IINS-AFP classification presents better prognostic performance than IINS or AFP alone, and might serve as a practical guidance to help patients adjust treatment and follow-up strategies to improve future outcomes.

Weighted gene co-expression network analysis reveals prognostic and diagnostic significance of PAQR4 in patients with early and late hepatocellular carcinoma.

Background: This study aimed to reveal novel markers for prognostic and diagnostic prediction of hepatocellular carcinoma (HCC). Methods: We applied The Cancer Genome Atlas (TCGA) data to screen differentially expressed genes (DEGs). We identified hub modules and genes using weighted gene co-expression network analysis (WGCNA). After verification with the GSE36376 dataset, hub genes were further identified. The expression of progestin and adipoQ receptor 4 (PAQR4) was confirmed in HCC by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The diagnostic and prognosis value of PAQR4 was assessed. The expression of PAQR4 was verified using the GSE76427 dataset. Results: A total of 803 DEGs were obtained between HCC and normal tissue. Through WGCNA, 7 hub modules were screened, among which the blue module was selected to identify the hub genes associated with the HCC. After overlapping all the DEGs with 837 genes of the blue module, we obtained 466 DEGs that were defined as hub genes. Among the hub genes, 239 were related to staging. After verifying with the GSE36376 dataset, PAQR4 was identified as the real hub gene of HCC. The results of qRT-PCR revealed that PAQR4 was upregulated between HCC and normal tissue. Furthermore, PAQR4 was related to the diagnosis and prognosis of patients with HCC. Moreover, the GSE76427 verification results of PAQR4 were consistent with our integration and qRT-PCR results. Ultimately, high expression of PAQR4 was significantly related to cell cycle, DNA replication, and the p53 signaling pathway. Conclusions: The PAQR4 gene may be associated with the prognosis and diagnosis of HCC.

Novel Therapeutic Mechanism of Adipose-Derived Mesenchymal Stem Cells in Osteoarthritis via Upregulation of BTG2.

Background: Osteoarthritis (OA) is a debilitating and degenerative joint disease, which is characterized by progressive destruction of articular cartilage. Mesenchymal stem cells (MSCs) have been implicated in the treatment of OA. However, the function of adipose-derived MSCs (AD-MSCs) in OA and its underlying mechanism remain obscure. Aim: We aimed to explore the function of AD-MSCs in OA and investigate its potential regulatory mechanism. Methods: A guinea pig model of OA was constructed. AD-MSCs injected into the articular cavity of OA guinea pigs were viewed by in vivo bioluminescence imaging. The effect of AD-MSCs on the gonarthritis of OA guinea pigs was evaluated through both macroscopic and microscopic detections. The detailed molecular mechanism was predicted by GEO databases and bioinformatics tools and then verified via mechanism experiments, including ChIP assay, DNA pulldown assay, and luciferase reporter assay. Results: AD-MSCs had a significant positive therapeutic effect on the gonarthritis of the OA model, and the overall effects of it was better than that of sodium hyaluronate (SH). B-cell translocation gene 2 (BTG2) was significantly downregulated in the articular cartilage of the OA guinea pigs. Furthermore, BTG2 was positively regulated by Krüppel-like factor 4 (KLF4) in AD-MSCs at the transcriptional level. AD-MSCs performed an effect on KLF4 expression at the transcriptional levels. Conclusion: AD-MSCs suppresses OA progression through KLF4-induced transcriptional activation of BTG2. Our findings revealed an AD-MSCs-dominated therapeutic method for OA.