Traditional Chinese medicine (Xielikang) reduces diarrhea symptoms in acquired immune deficiency syndrome (AIDS) patients by regulating the intestinal microbiota.

Diarrheal acquired immune deficiency syndrome (AIDS) seriously affects the quality of life of patients. In this study, we analyzed the differences in the intestinal microbiota among healthy individuals, AIDS patients without diarrhea and AIDS patients with diarrhea through high-throughput sequencing. The microbial diversity in the intestines of patients in the AIDS diarrhea group was significantly increased, and after treatment with Xielikang, the intestinal microbial diversity returned to the baseline level. At the phylum level, compared those in to the healthy (ZC) and AIDS non diarrhea (FN) groups, the relative abundances of Bacteroidetes and Verrucomirobia in the AIDS diarrhea (FA) group before treatment were significantly increased, while the relative abundance of Firmicutes was significantly decreased. Similarly, compared with those in the FA group, the relative abundances of Bacteroidea and Firmicutes in the AIDS diarrhea (FB) group after treatment were significantly increased, while the relative abundance of Firmicutes was significantly decreased after treatment. Additionally, there was no significant difference between the ZC and FN groups. At the genus level, compared with those in the ZC group, the relative abundance of Prevotella and Escherichia_Shigella in the FA group was significantly increased, while the relative abundances of Megamonas and Bifidobacterium was significantly decreased compared to that in the ZC group. After treatment with Xielikang, the relative abundance of Prevotella and Escherichia_Shigella in the FB group were significantly decreased, while the relative abundances of Megamonas and Bifidobacteria were significantly increased than those in the FA group; moreover, there was no significant difference between the ZC and FN groups. The functional prediction results showed that the ketodeoxyoctonate (Kdo) transfer to lipid IVA III and the superpathway of N-acetylglucosamine pathways in the AIDS diarrhea group were significantly altered. The correlation analysis results showed that Dorea was positively correlated with inflammatory factors, while Streptococcus and Lactobacillus were negatively correlated with inflammatory factors. The composition and function of the intestinal microbiota changed significantly in AIDS diarrhea patients, which affected the immune function of the host. The Xielikang capsule modulated the composition of the intestinal microbiota in AIDS diarrhea patients and thus improved immune function and reduced diarrheal symptoms.

Characterization of intestinal fungal community diversity in people living with HIV/AIDS (PLWHA).

Acquired Immune Deficiency Syndrome (AIDS) is a highly dangerous infectious disease caused by the Human Immunodeficiency Virus (HIV), a virus that attacks the human immune system. To explore the correlation between intestinal fungal community and immune function (Immune cells and inflammatory factors) in people living with HIV/AIDS (PLWHA). The feces and blood samples were collected from two groups of subjects: PLWHA and healthy controls. High-throughput sequencing of the internal transcribed spacer 1, flow cytometry, and ELISA were performed to analyze the differences and correlations between fungal microbiota, cellular immune status and serum inflammatory factors in the two groups. There were significant differences in the composition of fungal microbiota between the two groups. The relative abundance of Candida, Bjerkandera, and Xeromyces in PLWHA was significantly higher than that of healthy volunteers (P < 0.01), while the relative abundance of Mycospaerella, Xeroxysium, Penicillium, and Glomerella in PLWHA was significantly lower than that of healthy volunteers. The correlation analysis results show that Mycospaerella and Xeromyces are significantly positively correlated with CD4+/CD8+ T cells and the anti-inflammatory cytokine IL-4. On the other hand, Candida was positively correlated with pro-inflammatory factors negatively correlated with CD4+/CD8+ T cells and the anti-inflammatory cytokine IL-4, while it is positively correlated with pro-inflammatory cytokines. The significant increase in the relative abundance of Candida may be one of the important causes of intestinal damage in PLWHA. The results of this study contribute to the understanding of the relationship between fungal microbiota structure and immune function in the gut ecology of PLWHA.

Exosome-Transmitted miR-25 Induced by H. pylori Promotes Vascular Endothelial Cell Injury by Targeting KLF2.

Background: Increasing evidence has shown that Helicobacter pylori is associated with coronary heart disease (CHD); however, the underlying mechanism remains unclear. Methods: The expression of miR-25 and mRNAs was measured using qRT-PCR. Protein levels were detected using western blotting and exosomes were assessed with an electron microscope. The target gene of miR-25 was identified using the luciferase report system. Results:H. pylori infection increased the expression of miR-25 in gastric epithelial cells and was associated with increased levels of exosome-transmitted miR-25 in human peripheral blood. Mechanistic investigation showed the Kruppel-like factor 2 (KLF2) was a direct target of exosome-transmitted miR-25 in vascular endothelial cells. In addition, the miR-25/KLF2 axis regulated the NF-κB signaling pathway, resulting in increased expression of interleukin 6 (IL6), monocyte chemoattractant protein-1 (MCP-1), vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1). Conclusion: Our findings suggest that the miR-25/KLF2 axis may be a potential therapeutic target for H. pylori-associated CHD. Furthermore, high levels of exosome-transmitted miR-25 in peripheral blood may pose a potential risk for CHD.

NF-κB signaling and integrin-ß1 inhibition attenuates osteosarcoma metastasis via increased cell apoptosis.

Osteosarcoma is a common primary bone malignancy, and distant metastasis limited the cure estimate during last decades. Detailed investigation of osteosarcoma metastasis is valuable for improving therapeutic strategy. Our study indicated increased integrin-ß1 expression and NF-kB signaling activation in metastatic osteosarcoma tissues. Gain-of-function assays showed that integrin-ß1 knockdown significantly inhibited osteosarcoma growth and metastasis, whereas exogenous reintroducing of integrin-ß1 restored cell proliferation and metastasis in vitro and in vivo. NF-κB signaling directly modulated integrin-ß1 expression, which is an effective target for the treatment of osteosarcoma. Mechanically, integrin-ß1 blockage with AIIB2 antibody increased osteosarcoma cell apoptosis. Immunohistochemistry staining of integrin-ß1 revealed that elevated integrin-ß1 expression was correlated with poor prognosis of osteosarcoma patients and acted as an independent detrimental factor for osteosarcoma. Our data showed that integrin-ß1 and NF-κB signaling are promising therapeutic targets to improve the clinical outcome of osteosarcoma patients. The examination of integrin-ß1 expression will also identify patients with high risk of disease progression.

Partial Recovery of Limb Function Following End-to-Side Screw Anastomosis of Phrenic Nerve in Rats with Brachial Plexus Injury.

BACKGROUND Brachial plexus injury (BPI), a severe nervous system injury, is a leading cause of functional damages of the affected upper limb. Patients with BPI manifested with motor weakness or paralysis, sensory deficits, and pain. We established a BPI rat model to explore the in vivo effect of end-to-side screw anastomosis (ETSSA) of phrenic nerve on the recovery of limb function after BPI. MATERIAL AND METHODS After modeling, rats were treated with end-to-side anastomosis (ETSA) and ETSSA respectively. After 1 and 3 months, the behavioral changes of rats were observed using the Terzis grooming test, and the compound muscle action potential (CMAP) and muscle tension of biceps brachii were detected. The muscle weight recovery rate (MWRR) and cross-sectional area recovery rate (CARR) were calculated. Toluidine blue staining was used to observe the myelinated nerve fibers in the proximal phrenic nerve and distal musculocutaneous nerve of suture. The ratio of regenerated nerve traversing rate (NTR) was counted and motor endplate area of biceps brachii was measured. RESULTS The rats treated with ETSA and ETSSA exhibited elevated grading of Terzis grooming test with time. Although both the ETSSA and ETSA can reduce the MWRR, CARR and motor endplate area in BPI rats, ETSSA showed a better influence on the latency delayed rate (LDR) and amplitude recovery rate (ARR) of CMAP, muscular tension recovery rate (MTRR), MWRR, number of regenerated myelinated nerve fibers, NTR, and motor endplate area in BPI rats. CONCLUSIONS Our study provided evidence that ETSSA can restore the limb function recovery to a greater extent, and accelerate the regeneration of nerve fibers in rats with BPI; the effect of ETSSA was better than that of ETSA.

Vitamin K intake and the risk of fractures: A meta-analysis.

The association between dietary vitamin K intake and the risk of fractures is controversial. Therefore we perform a meta-analysis of cohort or nested case-control studies to investigate the relationship between dietary vitamin K intake and the risk of fractures. A comprehensive search of PubMed and EMBASE (to July 11, 2016) was performed to identify cohort or nested case-control studies providing quantitative estimates between dietary vitamin K intake and the risk of fractures. Summary relative risk (RRs) with corresponding 95% confidence intervals (CIs) were pooled by using a random-effects model. Four cohort studies and one nested case-control study, with a total of 1114 fractures cases and 80,982 participants, were included in our meta-analysis. Vitamin K intake in all included studies refers exclusively to the intake of phylloquinone (vitamin K1), which is the predominant form of vitamin K in foods. We observed a statistically significant inverse association between dietary vitamin K intake and risk of fractures (highest vs. the lowest intake, RR = 0.78, 95% CI: 0.56-0.99; I = 59.2%, P for heterogeneity = .04). Dose-response analysis indicated that the pooled RR of fracture for an increase of 50 µg dietary vitamin K intake per day was 0.97 (95% CI: 0.95-0.99) without heterogeneity among studies (I = 25.9%, P for heterogeneity = .25). When stratified by follow-up duration, the RR of fracture for dietary vitamin K intake was 0.76 (95% CI: 0.58-0.93) in studies with more than 10 years of follow-up. Our study suggests that higher dietary vitamin K intake may moderately decrease the risk of fractures.

A MicroRNA-124 Polymorphism is Associated with Fracture Healing via Modulating BMP6 Expression.

BACKGROUND: miR-124-3p has been reported to be involved in the pathogenesis of many diseases by modulating a variety of signaling pathways. In this study, we aimed to understand the impact of miR-124-3p expression level on the fracture healing in the patients of metaphyseal fracture of distal tibia, who received minimal invasive percutaneous plate osteosynthesis. METHODS: We firstly collected 195 patients of metaphyseal fracture of distal tibia, and the genotype of rs531564 was determined: GG (n=124) and GC+CC (n=71). We collected information of the participants including age, gender, total in-hospital time, smoking and alcohol consumption. Subsequently, we searched the miRNA database online to identify the possible binding sequence of miR-124-3p located within the 3'-UTR of the target gene. We did correlation analysis and luciferase to understand the regulatory relationship between miR-124-3p and BMP6. Meanwhile, we also conducted real time PCR and western blotting analysis to study the mRNA and protein expression level of BMP6 in different genotype groups. We then treated the cells with scramble control, miR-124-3p mimics, BMP6 siRNA and miR-124-3p inhibitors to investigate the influence of miR-124-3p on the expression of BMP6, viability and apoptosis of cells. RESULTS: Total in-hospital time was significantly longer in GC+CC group than GG group. MiR-124-3p was up-regulated in GG group than GC and CC groups. BMP6 was virtual target of miR-124-3p. There existed negative regulatory relationship betweenmiR-124-3p and BMP6. The mRNA and protein expression level of BMP6 decreased in GG group. MiR-124-3p decreased the expression of BMP6. MiR-124-3p negatively interfered with the viability of cells and BMP6 positively interfered with the viability of cells. MiR-124-3p reduced apoptosis and BMP6 promoted apoptosis. CONCLUSION: These data proved the expression of miR-124-3p was associated with the healing of metaphyseal fracture of distal tibia, and could be recognized as a biomarker to predict the healing of metaphyseal fracture of distal tibia.

Relationship between PI3K pathway and angiogenesis in CIA rat synovium.

To investigate the expression of hypoxia inducible factor (HIF-1α) and vascular endothelial growth factor (VEGF) in the synovium of collagen-induced arthritis (CIA) joint, and whether the PI3K pathway regulates angiogenesis in rheumatoid arthritis or not. A randomized controlled according to the principle of the rats were divided into normal control group (10 rats) and the experimental group (40 rats). The experimental group rats were established as type II collagen plus adjuvant Freund's complete adjuvant-induced arthritis model. HIF-1α and VEGF proteins' expression in serum of CIA rats group and normal control group were detected by ELISA. Microvessel density (MVD) in synovial tissue of CIA rats group and normal control group were detected by immunohistochemistry (IHC) staining. The protein expression of PTEN, PI3K, and AKT in synovial tissue were detected by Western Blot. Compared with normal control group, toes and ankle swelling and arthritis index (AI) of CIA rat increased, and the expression of VEGF and HIF-1α proteins in peripheral serum increased, IHC showed that MVD was significantly higher than that of the control group, and the difference was statistically significant (p<0.05). Western Blot results showed that PI3K and AKT proteins expression in CIA synovial tissue of rats increased, while the expression of PTEN protein decreased. Correlation analysis showed that VEGF and HIF-1 levels in the peripheral serum of CIA rats were positively correlated with arthritis index (AI); the contents of HIF-1α and VEGF in the peripheral serum of CIA rats were positively correlated with MVD in synovium tissue. The CIA rat model regulated the expression of HIF-1α and VEGF proteins in peripheral serum by PI3K signaling pathway, and then regulated neovascularization in RA.

Caveolin-1 is critical in the proliferative effect of leptin on osteoblasts through the activation of Akt.

Osteoblasts are critical in bone remodeling and the repair of bone fractures. Leptin is involved in bone metabolism and osteoblast survival through the downstream signaling pathway, however, the exact mechanism of the effect of leptin on osteoblasts remains to be fully elucidated. In the present study, hFOB 1.19 cells were used to observe the effects of leptin on cell proliferation and apoptosis, and to investigate the underlying mechanism. The results confirmed that treatment of hFOB 1.19 cells with leptin significantly induced cell proliferation. Western blot analysis showed that the expression of caveolin­1 and the activation of Akt in the cells treated with leptin were significantly increased, compared with the control cells. Additionally, inhibiting Akt activation eliminated the effects on cell proliferation induced by leptin. The rates of cell apoptosis and cell cycle distribution were examined using flow cytometry, which revealed a decrease in the apoptotic rate and an increase in the proportion of cells in the S phase. This indicated that leptin was capable of inducing cell proliferation by inhibiting apoptosis and stimulating cell progression to the S phase. Transfection of the cells with caveolin­1 small interfering RNA showed that the activation of Akt induced by leptin was significantly inhibited. Furthermore, caveolin­1 knockdown and inhibiting Akt activation eliminated the increased proliferation, increased proportion of cells in the S phase and increased anti­apoptotic effects induced by leptin. Taken together, the data obtained in the present study demonstrated that caveolin­1 was critical in the proliferative effect of leptin on osteoblasts via the activation of Akt.

Expression, regulation and function of MicroRNAs in endometriosis.

Endometriosis (EMS), characterized by the presence and growth of functional en do met rial-like tissues outside the uterine cavity, is a common and benign gyneco logical disorder with a poorly understood and somewhat enigmatic etiopathogenesis and pathophysiology. MicroRNAs (miRNAs) are single-stranded 19-25 nucleotide-long RNAs and have an important role in post-transcriptional gene silencing by base pairing with target mRNAs. Recent research has shown that miRNAs and their target mRNAs are differentially expressed in endometriosis and other disorders of the female reproductive system. In this paper, we review the recent progress in understanding the roles of miRNAs in endometriosis, and specific miRNAs as biomarkers and therapeutic targets for endometriosis.

Somatic copy number losses on chromosome 9q21.33q22.33 encompassing the PTCH1 loci associated with cardiac fibroma.

Cardiac fibroma is an extremely rare benign tumor that remains poorly characterized genetically. Somatic copy number alterations are common in tumors and have been defined as a crucial factor leading to tumors. In this study, we present a child diagnosed with cardiac fibroma with somatic copy number losses of a total of three discontinuous segments from 9q21.33 to 9q22.33, including a mosaic deletion of PTCH1. PTCH1 has been associated with sporadic cardiac fibroma. Sequencing analysis of the PTCH1 gene has not revealed any causative mutation. Quantitative PCR analysis of PTCH1 further confirms somatic copy number losses. Our data narrow down the critical causative deletions for sporadic cardiac fibroma to a region more precise than any other previously reported one. Our results suggest important roles of somatic copy number losses on chromosome 9q21.33q22.33 in the development of sporadic cardiac fibroma; these findings may provide a better understanding of sporadic cardiac fibroma pathogenesis and contribute to the identification of novel diagnostic biomarkers of this neoplasm. .

Minimally invasive unilateral versus bilateral technique in performing single-segment pedicle screw fixation and lumbar interbody fusion.

PURPOSE: The minimally invasive transforaminal lumbar interbody fusion procedure with percutaneous pedicle screws was adopted in clinical practice, but the choice between a unilateral pedicle screw (UPS) or bilateral pedicle screw (BPS) fixation after lumbar fusion remains controversial. The purpose of the present retrospective study was to compare the clinical outcomes and radiological results of unilateral and bilateral pedicle screw fixations. METHODS: The retrospective study recruited seventy-eight patients with a single-level pedicle screw fixation and lumbar interbody fusion at L4-L5 or L5-S1 from January 2010 to January 2013. The patients were treated with MIS TLIF with BPS fixation, and since May 2012, all patients were treated with UPS fixation. The perioperative outcomes including operative time, blood loss, hospital-stay length, and complication rates were accessed. Radiological outcomes regarding fusion were determined with the Bridwell grading system. Clinical outcomes were evaluated with the Oswestry Disability Index (ODI) and visual analog scale (VAS) during the mean follow-up of 2 years. RESULTS: According to perioperative assessments, the operative time was significantly shorter for group UPS (84.7 ± 6.4 min) than for group BPS (103.6 ± 10.6 min; p < 0.0001), and similar results were found with regard to the mean blood loss (UPS, 96.3 ± 17.5; BPS, 137.4 ± 32.9, p < 0.0001). With regard to the hospital-stay period, though the UPS group seems shorter, there is no statistical significance (UPS, 10.0 ± 2.1; BPS, 10.4 ± 2.4, p = 0.428). There were four in the BPS group and six in the UPS group defined as unfused at 6 months pest-operative, but at 12 months post-surgery, all patients achieved solid fusion. Regarding clinical outcomes, the VAS and ODI scores were significantly lower in the UPS group than the BPS group at 7 days post-surgery, but there was no difference at 1 month post-surgery and during the later follow-up. CONCLUSION: There was no difference between the UPS and BPS flexion techniques about the clinical outcomes at 24 months post-surgery. However, because the UPS involves a shorter surgical time, less blood loss, faster pain relief, and faster functional recovery, UPS might be more suitable in performing single-segment pedicle screw fixation and lumbar interbody fusion.

Safety of denosumab in postmenopausal women with osteoporosis or low bone mineral density: a meta-analysis.

PURPOSE: The aim of this meta-analysis was to assess the safety of denosumab in postmenopausal women with osteoporosis or low bone mineral density (BMD). METHODS: Safety of denosumab was compared with placebo or bisphosphonates. A systematic literature search without language restriction was conducted up to January, 2014. The RevMan 5.1 software was used for statistical analysis. RESULTS: A total of 11 English literatures were eventually identified. The pooled data in the overall analysis revealed that there was no significant difference when compared denosumab with placebo or bisphosphonates in any adverse events (AAE) (RR=0.99, 95% CI=0.98-1.01, p=0.29), serious adverse event (SAE) (RR=1.05, 95% CI=0.98-1.13, p=0.18), neoplasm/cancer (RR=1.14, 95% CI=0.95-1.37, p=0.16) and deaths (RR=0.77, 95% CI=0.57-1.04, p=0.09). However, significant differences were found when compared denosumab with placebo or bisphosphonates in SAE related to infection (RR=1.23, 95% CI=1.00-1.52, p=0.05) and non-vertebral fracture (RR=0.86, 95% CI=0.74-1.00, p=0.05). Subgroup analysis was performed by the type of drugs which was used in the control group. The results of subgroup analysis did not demonstrate the differences between denosumab and bisphosphonates in SAE related to infection (RR=1.13, 95% CI=0.63-2.03) and non-vertebral fracture (RR=1.31, 95% CI=0.87-1.98). CONCLUSIONS: Compared to placebo, denosumab treatment significantly decreased the risk of non-vertebral fracture but increased the risk of SAE related to infection in the postmenopausal women with osteoporosis or low BMD. However, no difference between the safety of denosumab and bisphosphonates was found.