Fecal Microbiota Transplantation Improves Clinical Symptoms of Fibromyalgia: An Open-Label, Randomized, Nonplacebo-Controlled Study.

Fibromyalgia (FM) is a complex and poorly understood disorder characterized by chronic and widespread musculoskeletal pain, of which the etiology remains unknown. Now, the disorder of the gut microbiome is considered as one of the main causes of FM. This study aimed to investigate the potential benefits of fecal microbiota transplantation (FMT) in patients with FM. A total of 45 patients completed this open-label, randomized, nonplacebo-controlled clinical study. The numerical rating scale scores in the FMT group were slightly lower than the control group at 1 month (P > .05), and they decreased significantly at 2, 3, 6, and 12 months after treatment (P < .001). Besides, compared with the control group, the Widespread Pain Index, Symptom Severity, Hospital Anxiety and Depression Scale, and Pittsburgh Sleep Quality Index scores were significantly lower in the FMT group at different time points (P < .001). After 6 months of treatment, there was a significant increase in serotonin (5-hydroxytryptamine) and gamma-aminobutyric acid levels (P < .001), while glutamate levels significantly decreased in the FMT group (P < .001). The total effective rate was higher in the FMT group (90.9%) compared to the control group (56.5%) after 6 months of treatment (P < .05). FMT can effectively improve the clinical symptoms of FM. With the close relations between the changes in neurotransmitters and FM, certain neurotransmitters may serve as a diagnostic marker or potential target for FM patients. PERSPECTIVE: FMT is a novel therapy that aims to restore the gut microbial balance and modulate the gut-brain axis. It is valuable to further explore the therapeutic effect of FMT on FM. Furthermore, certain neurotransmitters may become a diagnostic marker or a new therapeutic target for FM patients.

Long-term hypercaloric diet exacerbates metabolic liver disease in PNPLA3 I148M animals.

BACKGROUND & AIMS: Nonalcoholic fatty liver disease (NAFLD) is a major health burden associated with the metabolic syndrome leading to liver fibrosis, cirrhosis and ultimately liver cancer. In humans, the PNPLA3 I148M polymorphism of the phospholipase patatin-like phospholipid domain containing protein 3 (PNPLA3) has a well-documented impact on metabolic liver disease. In this study, we used a mouse model mimicking the human PNPLA3 I148M polymorphism in a long-term high fat diet (HFD) experiment to better define its role for NAFLD progression. METHODS: Male mice bearing wild-type Pnpla3 (Pnpla3WT ), or the human polymorphism PNPLA3 I148M (Pnpla3148M/M ) were subjected to HFD feeding for 24 and 52 weeks. Further analysis concerning basic phenotype, inflammation, proliferation and cell death, fibrosis and microbiota were performed in each time point. RESULTS: After 52 weeks HFD Pnpla3148M/M animals had more liver fibrosis, enhanced numbers of inflammatory cells as well as increased Kupffer cell activity. Increased hepatocyte cell turnover and ductular proliferation were evident in HFD Pnpla3148M/M livers. Microbiome diversity was decreased after HFD feeding, changes were influenced by HFD feeding (36%) and the PNPLA3 I148M genotype (12%). Pnpla3148M/M mice had more faecal bile acids. RNA-sequencing of liver tissue defined an HFD-associated signature, and a Pnpla3148M/M specific pattern, which suggests Kupffer cell and monocytes-derived macrophages as significant drivers of liver disease progression in Pnpla3148M/M animals. CONCLUSION: With long-term HFD feeding, mice with the PNPLA3 I148M genotype show exacerbated NAFLD. This finding is linked to PNPLA3 I148M-specific changes in microbiota composition and liver gene expression showing a stronger inflammatory response leading to enhanced liver fibrosis progression.

The Effects of Ginkgo biloba Extract on Autophagy in Human Macrophages Stimulated by Cigarette Smoke Extract.

OBJECTIVE: To investigate the effects of Ginkgo biloba extract (GBE) on autophagy in human macrophages stimulated by cigarette smoke extract (CSE). METHODS: The human monocyte cell line U937 was cultured in vitro, and phorbol ester (PMA) was added to the cell culture medium to induce differentiation into human macrophages. CSE was prepared by traditional methods for experiments. The cells were divided into four groups: the blank group, the CSE model group, the GBE + CSE group, and the rapamycin + CSE group. Immunofluorescence was used to identify human macrophages, transmission electron microscopy was used to observe the ultrastructure of human macrophages in each group, ELISA was used to measure the amount of IL-6 and IL-10 in the supernatant from each group of cells, the mRNA levels of p62, ATG5, ATG7, and Rab7 were measured by real-time qPCR, and the protein expression levels of p62, ATG5, ATG7, and Rab7 were measured by Western blotting. RESULTS: U937 cells were successfully differentiated into human macrophages after induction with PMA. The CSE model group had many more autophagosomes than the blank group. Compared with the CSE model group, the GBE + CSE group and the rapamycin + CSE group had significantly more autophagolysosomal. Compared with the other groups, the CSE model group had a higher level of IL-6 but a lower level of IL-10 in the supernatant (p < 0.05). Compared with the blank group, the mRNA and protein expression levels of p62 in the CSE model group were significantly decreased, while the mRNA and protein expression levels of ATG5 and ATG7 were significantly increased in the CSE model group (p < 0.05). No difference was found in the mRNA and protein expression levels of Rab7 between the blank group and the CSE model group. Compared with the CSE model group, the IL-6 level in the GBE + CSE group and the rapamycin + CSE group cell culture supernatant decreased significantly, p62 mRNA and protein expression significantly decreased, while ATG5, ATG7, and Rab7 mRNA and protein expression levels were significantly increased (p < 0.05). Moreover, increased LC3-II/LC3-I ratio were also found in the GBE + CSE group and the rapamycin + CSE group compared with the CSE model group. CONCLUSIONS: GBE could promote the fusion of autophagosomes and lysosomes in human macrophages, enhance the autophagy function of human macrophages, and reduce the damaging effect of CSE on the autophagy function of macrophages.

Intravenous Lidocaine Significantly Reduces the Propofol Dose in Elderly Patients Undergoing Gastroscopy: A Randomized Controlled Trial.

Objective: Propofol-based sedation has been widely used for gastroscopy, but the risk of respiratory suppression in elderly patients should not be overlooked. Intravenous (IV) lidocaine during surgery can reduce the demand for propofol and the incidence of cardiopulmonary complications. We examined whether IV lidocaine reduces the dose of propofol and the occurrence of adverse events during gastroscopy in elderly patients. Methods: We conducted a prospective, single-center, double-blind randomized controlled trial in elderly patients aged ≥65 years with ASA I-II. Subjects were randomly assigned to the lidocaine group (Group L, n=70), who received IV 1.5 mg kg-1 lidocaine followed by a continuous infusion of 4 mg kg-1 h-1 lidocaine, or the normal saline group (Group N, n=70), who received an equal volume of saline in the same way. Results: IV lidocaine reduced the total and maintenance propofol dose in Group L (p<0.001), with no significant effect on the induction dose. The incidence of intraoperative hypoxia (p=0.035), emergency airway management events (p=0.005), duration of gastroscopy (p<0.05), consciousness recovery time (p<0.001), and postoperative pain (p=0.009) were all reduced in Group L. Patient (p=0.025) and gastroscopist (p=0.031) satisfaction was higher in Group L. Intraoperative hemodynamic parameters, the respiratory rate, the incidence of sedation-related events and anesthesiologist satisfaction were similar between the two groups. Conclusion: IV lidocaine can significantly reduce the amount of propofol, the incidence of hypoxia and postoperative pain during gastroscopy in elderly patients, with a higher patient and gastroscopist satisfaction.

A MicroRNA-Based Network Provides Potential Predictive Signatures and Reveals the Crucial Role of PI3K/AKT Signaling for Hepatic Lineage Maturation.

BACKGROUND: Functions of miRNAs involved in tumorigenesis are well reported, yet, their roles in normal cell lineage commitment remain ambiguous. Here, we investigated a specific "transcription factor (TF)-miRNA-Target" regulatory network during the lineage maturation of biliary tree stem cells (BTSCs) into adult hepatocytes (hAHeps). METHOD: Bioinformatic analysis was conducted based on our RNA-seq and microRNA-seq datasets with four human hepatic-lineage cell lines, including hBTSCs, hepatic stem cells (hHpSCs), hepatoblasts (hHBs), and hAHeps. Short time-series expression miner (STEM) analysis was performed to reveal the time-dependent dynamically changed miRNAs and mRNAs. GO and KEGG analyses were applied to reveal the potential function of key miRNAs and mRNAs. Then, the miRDB, miRTarBase, TargetScan, miRWalk, and DIANA-microT-CDS databases were adopted to predict the potential targets of miRNAs while the TransmiR v2.0 database was used to obtain the experimentally supported TFs that regulate miRNAs. The TCGA, Kaplan-Meier Plotter, and human protein atlas (HPA) databases and more than 10 sequencing data, including bulk RNA-seq, microRNA-seq, and scRNA-seq data related to hepatic development or lineage reprogramming, were obtained from both our or other published studies for validation. RESULTS: STEM analysis showed that during the maturation from hBTSCs to hAHeps, 52 miRNAs were downwardly expressed and 928 mRNA were upwardly expressed. Enrichment analyses revealed that those 52 miRNAs acted as pluripotency regulators for stem cells and participated in various novel signaling pathways, including PI3K/AKT, MAPK, and etc., while 928 mRNAs played important roles in liver-functional metabolism. With an extensive sorting of those key miRNAs and mRNAs based on the target prediction results, 23 genes were obtained which not only functioned as the targets of 17 miRNAs but were considered critical for the hepatic lineage commitment. A "TF-miRNA-Target" regulatory network for hepatic lineage commitment was therefore established and had been well validated by various datasets. The network revealed that the PI3K/AKT pathway was gradually suppressed during the hepatic commitment. CONCLUSION: A total of 17 miRNAs act as suppressors during hepatic maturation mainly by regulating 23 targets and modulating the PI3K/AKT signaling pathway. The regulatory network uncovers possible signatures and guidelines enabling us to identify or obtain the functional hepatocytes for future study.

Ultrasound-Guided Continuous Thoracic Paravertebral Infusion of Methylene Blue in the Treatment of Postherpetic Neuralgia: A Prospective, Randomized, Controlled Study.

INTRODUCTION: Postherpetic neuralgia (PHN) is the most common complication of herpes zoster. Methylene blue (MB) is an inhibitor of nitric oxide synthesis with potentially analgesic and anti-inflammatory properties. Studies have demonstrated that thoracic paravertebral single MB injection is effective in treating chronic pain. However, there are rare reports of the efficacy of continuous thoracic paravertebral infusion of MB for pain management in PHN patients. The purpose of this study was to evaluate the therapeutic effects of continuous thoracic paravertebral infusion of MB on PHN. METHODS: A total of 104 PHN patients were randomly divided into two groups: the control group (continuous thoracic paravertebral infusion of 5% lidocaine in a total volume of 300 ml) and the MB group (continuous thoracic paravertebral infusion of 5% lidocaine plus 0.2% MB in a total volume of 300 ml). All patients were evaluated using the Numerical Rating Scale (NRS), Insomnia Severity Index (ISI), Patient Health Questionnaire-9 (PHQ-9), 36-Item Short-Form Health Survey (SF-36), and medication doses before and after the procedure. The effective treatment rate and adverse complications were recorded 6 months after the procedure. RESULTS: In both groups, the NRS scores, ISI scores, PHQ-9 scores, and rescue medication dosages were significantly decreased at different time points after treatment compared to baseline, while the SF-36 scores were evidently improved at different time points after treatment compared to baseline. Compared with the control group, the MB group had significantly reduced NRS scores, ISI scores, PHQ-9 scores, and rescue medication dosages at each observation time point. Furthermore, the SF-36 scores in the MB group were significantly higher than those in the control group at each observation time point. The total effective treatment rate of the MB group was higher than that of the control group 6 months after the procedure. No severe adverse complications were observed in either group. CONCLUSIONS: Ultrasound-guided continuous thoracic paravertebral infusion with MB is a safe and effective therapy for PHN. Continuous infusion with MB can significantly reduce pain intensity, improve pain-related depression, increase quality of life, and decrease the amount of rescue medicine with no serious adverse complications.

Xenogeneic native decellularized matrix carrying PPARγ activator RSG regulating macrophage polarization to promote ligament-to-bone regeneration.

Host immune response to tissue engineering tissues or organs directly determines the graft survival and the integration with host. Our and other previous studies have successfully regenerated the organs/tissues based on allogeneic native decellularized matrix (aNDM). But the very limited aNDM clinically hinders the artificial organs/tissues application to resolve the native organs/tissues loss with high incidence. However, the xenogeneic NDM will induce host immune rejection leading to the transplantation failure. This study constructed the xenogeneic (porcine) NDM (xNDM) which carried the immunoregulator Rosiglitazone (xNDM-RSG), a synthetic highly selective agonist of peroxisome proliferator-activated receptor-γ (PPARγ), evaluated xNDM's physical and chemical characterization, immunomodulatory properties, and its effect on the tissue regeneration. Results showed that the xNDM-RSG did not affect the proliferation and differentiation of odontogenic stem cells. In addition, the xNDM-RSG could also effectively decrease the expression of IL-1 and TNFα, and increase the expression of IL-10 and TGFß to enable a favorable immunomodulation and promote the ligament-to-bone regeneration by PPARγ to induce the alternative activated macrophages (M2 macrophages) antagonizing classically activated macrophages (M1 macrophages). Meanwhile the xNDM-RSG obviously reduces the implants absorption and promotes the regenerated ligament-to-bone expressing the key proteins (ALP, OPN, DSP) which are relative to the native dental and bone. This study demonstrated that protein adsorption could aggravate the immune inflammatory reaction, whereas, xNDM-RSG could effectively control the host immune response to accelerate tissue reparation and regeneration by facilitating the macrophage polarization, which highlighted a new strategy for improving the transplantation survival of the artificial organ or tissue based on the xenogeneic decellularized biomaterials.

Parathyroid hormone-related peptide (1-34) promotes tooth eruption and inhibits osteogenesis of dental follicle cells during tooth development.

Dental follicle cells (DFCs) activate and recruit osteoclasts for tooth development and tooth eruption, whereas DFCs themselves differentiate into osteoblasts to form alveolar bone surrounding tooth roots through the interaction with Hertwig's epithelial root sheath (HERS). Also during tooth development, parathyroid hormone-related peptide (PTHrP) is expressed surrounding the tooth germ. Thus, we aimed to investigate the effect of PTHrP (1-34) on bone resorption and osteogenesis of DFCs in vitro and in vivo. In vitro studies demonstrated that DFCs cocultured with HERS cells expressed higher levels of BSP and OPN than the DFCs control group, whereas cocultured DFCs treated with PTHrP (1-34) had lower expressions of ALP, RUNX2, BSP, and OPN than the cocultured DFCs control group. Moreover, we found PTHrP (1-34) inhibited osteogenesis of cocultured DFCs by inactivating the Wnt/ß-catenin pathway. PTHrP (1-34) also increased the expression of RANKL/OPG ratio in DFCs. Consistently, in vivo study found that PTHrP (1-34) accelerated tooth eruption and inhibited alveolar bone formation. Therefore, these results suggest that PTHrP (1-34) accelerates tooth eruption and inhibits osteogenesis of DFCs by inactivating Wnt/ß-catenin pathway.

Inactivation of caspase 8 in liver parenchymal cells confers protection against murine obstructive cholestasis.

BACKGROUND & AIMS: Caspase 8 (CASP8) is the apical initiator caspase in death receptor-mediated apoptosis. Strong evidence for a link between death receptor signaling pathways and cholestasis has recently emerged. Herein, we investigated the role of CASP8-dependent and independent pathways during experimental cholestasis. METHODS: Liver injury was characterized in a cohort of human sera (n = 28) and biopsies from patients with stage IV primary biliary cholangitis. In parallel, mice with either specific deletion of Casp8 in liver parenchymal cells (Casp8Δhepa) or hepatocytes (Casp8Δhep), and mice with constitutive Ripk3 (Ripk3-/-) deletion, were subjected to surgical ligation of the common bile duct (BDL) from 2 to 28 days. Floxed (Casp8fl/fl) and Ripk3+/+ mice were used as controls. Moreover, the pan-caspase inhibitor IDN-7314 was used, and cell death mechanisms were studied in primary isolated hepatocytes. RESULTS: Overexpression of activated caspase 3, CASP8 and RIPK3 was characteristic of liver explants from patients with primary biliary cholangitis. Twenty-eight days after BDL, Casp8Δhepamice showed decreased necrotic foci, serum aminotransferase levels and apoptosis along with diminished compensatory proliferation and ductular reaction. These results correlated with a decreased inflammatory profile and ameliorated liver fibrogenesis. A similar phenotype was observed in Ripk3-/- mice. IDN-7314 treatment decreased CASP8 levels but failed to prevent BDL-induced cholestasis, independently of CASP8 in hepatocytes. CONCLUSION: These findings show that intervention against CASP8 in liver parenchymal cells - specifically in cholangiocytes - might be a beneficial option for treating obstructive cholestasis, while broad pan-caspase inhibition might trigger undesirable side effects. LAY SUMMARY: Loss of caspase 8 - a protein involved in programmed cell death - in liver parenchymal cells protects against experimental cholestasis. Therefore, specific pharmacological intervention against caspase 8 might be a valid alternative for the treatment of obstructive cholestasis in the clinic, whereas broad pan-caspase inhibiting drugs might trigger undesirable side effects.

CuCo2S4 nanocrystals: a new platform for multimodal imaging guided photothermal therapy.

Multifunctional nanomaterials open an avenue for the integration of cancer treatment and diagnosis, trending towards the clinical application of nanomedicines in future. Herein, we synthesized biocompatible CuCo2S4 nanocrystals (NCs) via a simple reflux method and unitized them as a new theranostic platform, where an intense near-infrared (NIR) absorption offers the CuCo2S4 NCs a perfect photothermal performance and photoacoustic (PA) imaging ability; the magnetic characteristic of Co allows them to serve as an enhanced magnetic resonance (MR) contrast agent. The in vitro and in vivo experiments demonstrated their good biocompatibility, non-toxicity and perfect photothermal conversion performance and further confirmed that HeLa tumors could be effectively thermal-ablated upon the assistance of the CuCo2S4 NCs. This work introduces the first bioapplication of the CuCo2S4 NCs and promotes theranostics based on other ternary compounds.

CX3CR1 is a gatekeeper for intestinal barrier integrity in mice: Limiting steatohepatitis by maintaining intestinal homeostasis.

UNLABELLED: Nonalcoholic fatty liver disease is seen as the hepatic manifestation of the metabolic syndrome and represents the most common liver disease in Western societies. The G protein-coupled chemokine receptor CX3CR1 plays a central role in several metabolic syndrome-related disease manifestations and is involved in maintaining intestinal homeostasis. Because diet-induced intestinal dysbiosis is a driver for nonalcoholic fatty liver disease, we hypothesized that CX3CR1 may influence the development of steatohepatitis. In two independent models of diet-induced steatohepatitis (high-fat diet and methionine/choline-deficient diet), CX3CR1 protected mice from excessive hepatic steatosis and inflammation, as well as systemic glucose intolerance. Lack of Cx3cr1 expression was associated with significantly altered intestinal microbiota composition, which was linked to an impaired intestinal barrier. Concomitantly, endotoxin levels in portal serum and inflammatory macrophages in liver were increased in Cx3cr1-/- mice, indicating an increased inflammatory response. Depletion of intestinal microbiota by administration of broad-spectrum antibiotics suppressed the number of infiltrating macrophages and promoted macrophage polarization in liver. Consequently, antibiotic-treated mice demonstrated a marked improvement of steatohepatitis. CONCLUSION: Microbiota-mediated activation of the innate immune responses through CX3CR1 is crucial for controlling steatohepatitis progression, which recognizes CX3CR1 as an essential gatekeeper in this scenario.

Correction of asymmetric facial deformity by contouring: indications and outcomes.

BACKGROUND: Facial asymmetry is usually due to unbalanced development of the lower jaw and zygoma and often results in esthetically unpleasant appearance. However, reasonable and systematic treatment of such an asymmetric face is rarely reported in the literature. This article aims to evaluate the effectiveness of surgical correction of asymmetric facial deformity and discuss their indications. METHODS: From July 2006 to November 2010, a total of 52 patients received contour reshaping procedures to correct their asymmetric faces. Those patients in whom the asymmetric facial deformities were initiated by hypertrophy of the mandible and zygoma without occlusion and temporomandibular joint problem were chosen for this study. The authors performed a modified reduction malarplasty to correct asymmetric middle face and mandibular outer cortex splitting ostectomy, mandibular "V-line" ostectomy, and rotation genioplasty to improve asymmetric lower face depending on individual asymmetric facial characteristics. The effectiveness was then evaluated through cephalometric radiographs, three-dimensional computed tomography, and presurgical and postsurgical standard facial photographs. RESULTS: The postoperative results of all 52 cases showed that the asymmetric face was effectively corrected without serious complications and the harmonious facial contour improved significantly. The final esthetic outcomes were quite satisfactory for both surgeons and patients. CONCLUSIONS: The results indicate that a variety of contouring techniques for facial asymmetric deformity could be carried out based on characteristics of asymmetric face, so as to acquire a symmetric and harmonious face in accordance with facial esthetics.

[Clinical observation of CT guided two needles puncturing crossed through disc for superior hypogastric block to manage intractable pelvic cancer pain].

OBJECTIVE: To observe the feasibility and clinical efficacy of CT guided two needles puncturing crossed through disc for superior hypogastric block with alcohol to manage intractable pelvic cancer pain. METHODS: Thirty-one cases of advanced pelvic cancer suffering from untreatable pain in lower abdomen in our hospital from December 2009 to May 2012 were analyzed, the patients were treated with both sides of superior hypogastric block with absolute alcohol by CT guided two needles puncturing crossed through L5-S1 interlaminar space. Complications during and after the surgery were recorded. To observe and follow-up visual analog scale pain scores (VAS) and the daily oral morphine consumption on just before operation, at 1 week, 1 and 3 months after operation. RESULTS: No case suffered serious complication. A week later of surgery the curative effect of 20 patients: clinical cure in 17 cases, excellent in 14 cases, the effective rate was 100%. Compared with preoperation, The score of VAS on 1 week, 1 and 3 months after the surgery (2.0±0.7, 2.3±0.6, 3.0±0.4) were strikingly lower than before operation (7.7±0.7, P < 0.01); The daily oral dose of morphine of post-operation were significantly decreased in the three time points ((35±17) mg, (42±22) mg and (53±19) mg respectively) than the dose of pre-operation ((201±119)mg, P < 0.01). CONCLUSION: Superior hypogastric block with alcohol with double needles crossed transdiscal approach may be a safe, simple and effective method for relieving the severe pain of advanced pelvic cancer patients.

[Design and biomechanical analysis of nickel-titanium open shape memory alloy artificial vertebral body].

OBJECTIVE: To design an open shape memory alloy artificial vertebral body that can be used to reconstruct the vertebral body in spine diseases, such as thoracic-lumbar spine tumors, burst fracture of the vertebrae, kyphosis and scoliosis, and to evaluate the biomechanical stability of lumbar functional segment unit after insertion with the shape memory alloy artificial vertebral body. METHODS: The open shape memory alloy artificial vertebral body with nickel-titanium (NiTi) alloy was made. Eight fresh spine specimens (T14-L5) from normal adult porcine were used to detect the range of motion (ROM) in 4 models and were divided into 4 groups: intact vertebrae served as group A; pedicle screw fixation of T15, L1, L3, and L4 was given in group B; after total resection of L2, it was reconstructed by open shape memory alloy artificial vertebral body combined with pedicle screw fixation of T15, L1, L3, and L4 in group C; and after total resection of L2, it was reconstructed by titanium cage vertebral body combined with pedicle screw fixation of T15, L1, L3, and L4 in group D. The three-dimensional ROM of flexion, extension, left/right lateral bending, and left/right rotation in T15-L1, L1-3, and L3,4 segments were detected in turn by the spinal three-dimensional test machine MTS-858 (load 0-8 N x m). RESULTS: Compared with group A, groups B, C, and D had good stability in flexion, extension, left/right lateral bending, and left/right rotation, showing significant differences (P < 0.05). There was no significant difference in the degree of each motion between group B and group C (P > 0.05). Group C had less degree of motion in T15-L1 and L3,4 segments than group D, showing significant differences (P < 0.05), but there was no significant difference in L1-3 segment (P > 0.05). CONCLUSION: The open shape memory alloy artificial vertebral body has a reasonable structure and good biomechanical stability, it can be used to stabilize the spinal segment with pedicle screw fixation.