Levetiracetam attenuates diabetes-associated cognitive impairment and microglia polarization by suppressing neuroinflammation.

Introduction: Cognitive impairment is a common complication and comorbidity of diabetes. However, the underlying mechanisms of diabetes-associated cognitive dysfunction are currently unclear. M1 microglia secretes pro-inflammatory factors and can be marked by CD16, iNOS, Iba1 and TNF-ɑ. The decline of M2 microglia in the diabetic rats indicates that high glucose promotes the differentiation of microglia into the M1 type to trigger neuroinflammatory responses. Moreover, there is a lack of strong evidence for treatments of diabetes-associated cognitive impairment in addition to controlling blood glucose. Methods: Diabetic rats were established by intraperitoneal injection of one dose of streptozotocin (60 mg/kg). Polarization transitions of microglia were induced by high glucose treatment in BV2 cells. Levetiracetam was orally administered to rats 72 h after streptozotocin injection for 12 weeks. Results: In STZ-induced diabetic rats, the results demonstrated that levetiracetam improved rat cognitive function (Morris water maze test) and hippocampus morphology (Hematoxylin-eosin staining), and the effect was more evident in the high-dose levetiracetam group. Microglia activation in the hippocampus was inhibited by levetiracetam treatment for 12 weeks. Serum levels of TNF-α, IL-1ß, and IL-6 were reduced in the LEV-L and LEV-H groups, and IL-1ß level was obviously reduced in the LEV-H group. In vitro, we found that levetiracetam 50 µM attenuated high-glucose induced microglial polarization by increasing IL-10 level and decreasing IL-1ß and TNF-α levels. Moreover, levetiracetam 50 µM increased and decreased the proportion of CD206+/Iba1+ and iNOS+/Iba1+cells, respectively. Western blot analysis illustrated that LEV 50 µM downregulated the expression of MyD88 and TRAF6, and phosphorylation of TAK1, JNK, p38, and NF-κB p65. The effect of levetiracetam on the anti-polarization and expression of p-JNK and p-NF-κB p65 were partly reversed by anisomycin (p38 and JNK activators). Discussion: Together, our data suggest that levetiracetam attenuates streptozotocin-induced cognitive impairment by suppressing microglia activation. The in vitro findings also indicate that the levetiracetam inhibited the polarization of microglia via the JNK/MAPK/NF-κB signaling pathway.

Hypothermia or hyperthermia, which is associated with patient outcomes in critically ill children with sepsis? -A retrospective study.

OBJECTIVES: In the early stage of sepsis, identifying high-risk paediatric patients with a poor prognosis and providing timely and adequate treatment are critical. This study aimed to evaluate the effect of average body temperature within 24 hours of admission on the short-term prognosis of paediatric patients with sepsis. DESIGN: A retrospective cohort study. SETTING: A single-centre, tertiary care hospital in China, containing patient data from 2010 to 2018. PARTICIPANTS: 1144 patients with sepsis were included. INTERVENTION: None. PRIMARY AND SECONDARY OUTCOME MEASURES: The main outcome measure was in-hospital mortality, which was defined as death from any cause during hospitalisation. The secondary outcome was the length of hospital stay. RESULTS: The LOWESS method showed a roughly 'U'-shaped relationship between body temperature on the first day and in-hospital mortality. Multivariate logistic regression showed that severe hypothermia (OR 14.72, 95% CI 4.84 to 44.75), mild hypothermia (OR 3.71, 95% CI 1.26 to 10.90), mild hyperthermia (OR 3.41, 95% CI 1.17 to 9.90) and severe hyperthermia (OR 5.15, 95% CI 1.84 to 14.43) were independent risk factors for in-hospital mortality. Compared with other variables, the Wald χ2 value of temperature on the first day minus the degree of freedom was the highest. CONCLUSIONS: Whether hypothermic or hyperthermic, the more abnormal the temperature on the first day is, the higher the risk of in-hospital death in children with sepsis.

P-Rex1 Cooperates With TGFßR2 to Drive Lung Fibroblast Migration in Pulmonary Fibrosis.

Pulmonary fibrosis is a kind of interstitial lung disease with progressive pulmonary scar formation, leading to irreversible loss of lung functions. The TGF-ß1/Smad signaling pathway plays a key role in fibrogenic processes. It is associated with the increased synthesis of extracellular matrix, enhanced proliferation of fibroblasts, and transformation of alveolar epithelial cells into interstitial cells. We investigated P-Rex1, a PIP3-Gßγ-dependent guanine nucleotide exchange factor (GEF) for Rac, for its potential role in TGF-ß1-induced pulmonary fibrosis. A high expression level of P-Rex1 was identified in the lung tissue of patients with pulmonary fibrosis than that from healthy donors. Using the P-Rex1 knockdown and overexpression system, we established a novel player of P-Rex1 in mouse lung fibroblast migration. P-Rex1 contributed to fibrogenic processes in lung fibroblasts by targeting the TGF-ß type Ⅱ receptor (TGFßR2). The RNA-seq analysis for expression profiling confirmed the modulation of P-Rex1 in cell migration and the involvement of P-Rex1 in TGF-ß1 signaling. These results identified P-Rex1 as a signaling molecule involved in TGF-ß1-induced pulmonary fibrosis, suggesting that P-Rex1 may be a potential target for pulmonary fibrosis treatment.

Identification of P-Rex1 in the Regulation of Liver Cancer Cell Proliferation and Migration via HGF/c-Met/Akt Pathway.

BACKGROUND: Rho-GTPases and their activators, guanine nucleotide exchange factors (GEFs), are increasingly being recognized as essential mediators of oncogenic signaling. Although it is known that P-Rex1, a member of the Dbl family of GEFs for the Rac small GTPase, contributes to the migration of cancer cells, its exact role in liver cancer and the underlying mechanisms remain unclear. MATERIALS AND METHODS: Public datasets from the Gene Expression Omnibus database (GEO) and clinical liver cancer samples were analyzed to explore the expression of P-Rex1. P-Rex1 knockdown and overexpression cell lines were established using a recombinant lentiviral transfection system. BrdU and colony formation assays were performed to determine cell viability. Migratory capacity was analyzed using a transwell migration assay and an in vitro wound-healing assay. Nude mice bearing subcutaneous xenograft tumors were established to determine the effects of P-Rex1 on tumorigenesis in vivo. The role of P-Rex1 in hepatocarcinogenesis was determined through Western blot and co-immunoprecipitation. RESULTS: Induced expression of endogenous P-Rex1 was identified in liver cancer tumors when compared with adjacent nonmalignant tissues from clinical data. In response to HGF treatment, P-Rex1-knockdown cells displayed reduced proliferation and migration in vitro as well as reduced xenograft tumor growth in vivo. Overexpression of P-Rex1 promoted liver cancer cell proliferation and migration. P-Rex1 primarily acts as a downstream effector of GPCR signaling. This study demonstrated that downregulation of P-Rex1 led to a significant decrease in the phosphorylation of Akt and Erk1/2 by reducing the phosphorylation of the tyrosine kinase receptor c-Met. Furthermore, a physical association between P-Rex1 and c-Met was observed after HGF treatment, suggesting that P-Rex1 may be involved in the HGF/c-Met signaling pathway. CONCLUSION: These results support the role of P-Rex1 as a novel player in liver cancer, which suggest that targeting P-Rex1 may provide a potential strategy for liver cancer treatment.

Ganoderic acid A alleviates hypoxia-induced apoptosis, autophagy, and inflammation in rat neural stem cells through the PI3K/AKT/mTOR pathways.

Effects of ganoderic acid A (GAA), a lanostane triterpene, on hypoxia-ischemia encephalopathy (HIE) remain unclear. We aimed to figure out the specific role of GAA in hypoxia-treated neural stem cells (NSCs) as well as the regulatory mechanisms. Primary rat NSCs were incubated under hypoxia to simulate HIE. Viability and apoptosis of hypoxia-injured NSCs were measured by cell counting kit-8 and flow cytometry assays, respectively. Proteins related to apoptosis, autophagy, and the PI3K/AKT/mTOR pathways were evaluated by Western blot analysis. LY294002 and rapamycin were added to inhibit the PI3K/AKT pathway and mTOR pathway, respectively. Enzyme-linked immunosorbent assay was carried out to test the release of proinflammatory cytokines. We found that hypoxia-induced decrease of cell viability, increases of apoptotic cells and autophagy, and the release of IL-6, IL-1ß, and TNF-α were all attenuated by GAA stimulation. Activation of caspases induced by hypoxia was alleviated by GAA. Furthermore, we found that inhibition of the PI3K/AKT pathway eliminated the effects of GAA on apoptosis and proinflammatory cytokines release in hypoxia-injured NSCs. Meanwhile, inhibition of the mTOR pathway abrogated the effects of GAA on cell autophagy in hypoxia-injured NSCs. In conclusion, GAA alleviated hypoxia-induced injury in NSCs might be through activating the PI3K/AKT and mTOR pathways.

Ginkgo biloba Extract Protects Mesenteric Arterioles of Old Rats via Improving Vessel Elasticity through Akt/FoxO3a Signaling Pathway.

BACKGROUND: Previous studies have shown that Ginkgo biloba extract (GBE) dietary diminished salt-related elevation of blood pressure and ameliorated ischemic diseases. However, whether GBE could improve vascular elasticity to protect mesenteric arterioles of old rats is still elusive. In this study, we aimed to investigate the effects of GBE on vascular elasticity of old rats and its possible underlying mechanism. METHODS: Morphological changes of mesenteric arterioles were observed using hematoxylin and eosin and Verhoeff-Van Gieson staining, and diameters of mesenteric arterioles under various pressure were detected after GBE administration. In addition, phosphorylation level of Akt and FoxO3a proteins from mesenteric arterioles were detected. RESULTS: The results implicated that GBE treatment narrowed endothelial cell gap and increased the curvature of inner elastic membrane with reduced middle layer collagen fiber. Meanwhile, compared with young rats, old rats appeared to have lower vascular elasticity while GBE treatment at 50, 100, and 200 mg/kg dosage through intragastric administration per day for 3 weeks could effectively improve the vascular elasticity under different pressures in a dose-dependent manner. Furthermore, phosphorylation level of Akt and FoxO3a was also reduced in GBE-treated rats. CONCLUSIONS: This is the first report to indicate that GBE might exert protective effect on mesenteric arterioles of old rats via improving vascular elasticity and Akt/FoxO3a signaling pathway might be involved in this action.

Transurethral balloon dilatation of the Prostate and Transurethral Plasmakinetic resection of the Prostate in the treatment of Prostatic Hyperplasia.

BACKGROUND & OBJECTIVE: With the aggravation of global aging, benign prostate hyperplasia tends to have a higher incidence and has been the most common disease in urinary surgery. It is usually treated by surgery. Our objective was to select an effective treatment scheme, the clinical efficacy and relevant indicators of transurethral balloon dilatation of the prostate (TUDP) and transurethral plasmakinetic resection of the prostate (PKRP) in the treatment of benign prostate hyperplasia were emphatically compared. METHODS: Ninety-eight patients with benign prostate hyperplasia who were admitted to the hospital of between May 2014 and July 2016 were selected and divided into a TUDP group (n=49) and PKRP (n=49) using random number table. The intraoperative blood loss, duration of surgery, international prostate symptom score (IPSS), quality of life (QOL), post-void residual urine (PVR) and complications of the two groups were observed. RESULTS: The results demonstrated that the postoperative blood loss and duration of surgery of the patients in the PKRP group were significantly higher than those of the TUDP group (P<0.05); the IPSS, QOL and PVR of the patients in the two groups after surgery were much lower than those before surgery (P<0.05); the IPSS, QOL and PVR of the patients in the PKRP group were significantly lower than those in the TUDP group after surgery (P<0.05). The incidence of postoperative complications of the PKRP group was 38.8%, which was apparently higher than 14.3% in the TUDP group (P<0.05). CONCLUSION: PKRP has better efficacy than TUDP in treating benign prostatic hyperplasia, but QOL was poor and there are many complications. Proper surgical procedure should be selected according to the specific disease condition of patients.

Tamsulosin and Solifenacin in the Treatment of Benign Prostatic Hyperplasia in combination with overactive bladder.

OBJECTIVE: To analyze the clinical effect of tamsulosin and Solifenacin in the treatment of benign prostatic hyperplasia in combination with overactive bladder and its safety. Another objective was to investigate the clinical effect and safety of mega dose of tamsulosin in the treatment of benign prostatic hyperplasia in combination with overactive bladder. METHODS: One hundred and twenty-four patients who were admitted to the Dept. of Urology at Binzhou People's Hospital, , China with confirmed benign prostatic hyperplasia (BPH) with overactive bladder were randomly divided into two groups. Sixty-two patients in the control group were treated with tamsulosin, while sixty-two patients in the observation group were treated with tamsulosin in combination with solifenacin. The treatment of both groups lasted for 12 weeks. The effect and adverse reaction were compared between the two groups. RESULTS: The international prostate symptom score (IPSS), quality of life (QOL), and overactive bladder symptom score (OABSS), Qmax, pulmonary vascular resistance (PVR), daytime urination frequency, urgent urination frequency, urge urinary incontinence frequency and night urinary frequency of both groups improved after treatment, and the difference had statistical significance (P<0.05). The differences of the observation indexes (except PVR) in the observation group before and after treatment was significantly different with those of the control group (P<0.05). The incidence of adverse reactions in the observation group was lower than that in the control group, but the difference had no statistical significance (X2=2.843, P>0.05). CONCLUSION: Treating benign prostatic hyperplasia in combination with overactive bladder with tamsulosin in combination with solifenacin is more effective than tamsulosin, without significantly increasing adverse reactions. Thus the therapy is worth clinical promotion.

Shikonin suppresses pulmonary fibroblasts proliferation and activation by regulating Akt and p38 MAPK signaling pathways.

Fibroblast is believed to be the primary effector in idiopathic pulmonary fibrosis (IPF), a progressive lung disorder characterized by aberrant tissue remodeling and the formation of fibroblastic foci. Due to the complicated etiology and mechanism, there are few effective drugs for this fatal disease. Shikonin (SHI), which is the major ingredient isolated from the plant Lithospermum Erythrorhizon, has long been used as traditional medicine for many diseases including inflammation and cancer. The roles of SHI in attenuating skin scar and renal fibrosis by reducing TGFß1-stimulated fibroblast activation are also reported. But whether SHI works on IPF which exhibits both inflammatory and carcinoma-like features remains unknown. In this study, using isolated pulmonary fibroblasts, we demonstrated that SHI inhibited the proliferation, migration of fibroblasts, enhanced cell apoptosis and led to cell cycle arrest at G1 and G2/M phase. Moreover, SHI reduced the production of α-SMA, fibronectin, collagen I and III in response to TGF-ß induction in pulmonary fibroblasts, and all of these gene production is the key component of extracellular matrix for tissue remodeling for IPF. The phosphorylation of Akt was down-regulated, p53 increased, the mRNA levels of p21 and p27 enhanced after SHI treatments. The phosphorylation of both p38 MAPK and Akt stimulated by TGF-ß was reduced after SHI treatments. Collectively, these data indicate that SHI has a strong cytotoxicity in pulmonary fibroblast via inhibiting Akt activation signaling pathway, and attenuates TGF-ß induced extracellular matrix genes production in pulmonary fibroblasts via modulating the activities of p38 MAPK and Akt. SHI might serve as a therapeutically candidate for IPF patients.

α-Solanine reverses pulmonary vascular remodeling and vascular angiogenesis in experimental pulmonary artery hypertension.

OBJECTIVE: Similar to cancer, pulmonary arterial hypertension (PAH) is characterized by vascular remodeling, which leads to obliteration of the small pulmonary arteriole, with marked proliferation of pulmonary artery smooth muscle cells (PASMC) and/or endothelial cells dysfunction. Aberrant expression of tumor suppressor genes is closely associated with susceptibility to PAH. We hypothesized that α-solanine, a glycoalkaloid found in members of the nightshade family known to have antitumor activity in different cancers, reverses experimental PAH by activating the tumor suppressor-axis inhibition protein 2 (AXIN2). METHODS AND RESULTS: We investigated the effects of α-solanine on PASMC proliferation and apoptosis by using 5-ethynyl-2'-deoxyuridine proliferation assay, proliferating cell nuclear antigen and Ki67 staining, TUNEL and Anexine V assays. Scratch wound healing and tube formation assays were also used to study migration of endothelial cells. In vitro, we demonstrated, using cultured human PASMC from PAH patients, that α-solanine reversed dysfunctional AXIN2, ß-catenin and bone morphogenetic protein receptor type-2 signaling, whereas restored [Ca]i, IL-6 and IL-8, contributing to the decrease of PAH-PASMC proliferation and resistance to apoptosis. Meanwhile, α-solanine inhibits proliferation, migration and tube formation of PAH-pulmonary artery endothelial cells by inhibiting Akt/GSK-3α activation. In vivo, α-solanine administration decreases distal pulmonary arteries remodeling, mean pulmonary arteries pressure and right ventricular hypertrophy in both monocrotaline-induced and Sugen/hypoxia-induced PAH in mice. CONCLUSION: This study demonstrates that AXIN2/ß-catenin axis and Akt pathway can be therapeutically targeted by α-solanine in PAH. α-Solanine could be used as a new therapeutic strategy for the treatment of PAH.

AKT2 Regulates Pulmonary Inflammation and Fibrosis via Modulating Macrophage Activation.

Idiopathic pulmonary fibrosis (IPF) is a highly lethal pathological process that is characterized by inflammation, fibroblast accumulation, and excessive collagen deposition. Although AKT2-mediated signaling pathways modulate inflammatory responses, their role in IPF has not been defined. We report that AKT2 deficiency (Akt2-/-) protected against bleomycin-induced pulmonary fibrosis and inflammation. Adoptive transfer of wild-type macrophages or administration of IL-13 to Akt2-/- mice could restore pulmonary fibrosis. In response to IL-33 treatment, Akt2-/- macrophages displayed decreased production of IL-13 and TGF-ß1 and attenuated phosphorylation of FoxO3a compared with Akt2+/+ macrophages. Furthermore, the expression of IL-13 was increased by small interfering RNA knockdown of FoxO3a or in FoxO3a-deficient macrophages. By evaluating lung sections from pulmonary fibrosis patients, we found that the phosphorylation of AKT2 and FoxO3a was remarkably upregulated. Collectively, these results indicate that AKT2 modulates pulmonary fibrosis through inducing TGF-ß1 and IL-13 production by macrophages, and inhibition of AKT2 may be a potential strategy for treating IPF.

Axis inhibition protein 2 deficiency leads to hypoxic pulmonary hypertension through ß-catenin signaling pathway.

OBJECTIVE: Pulmonary arterial hypertension (PAH) is characterized by increased vascular tone, altered vasoreactivity and vascular remodeling induced by smooth muscle cell proliferation. Similarities exist between cancer and PAH. Aberrant expression of the tumor suppressor protein is closely associated with PAH. Here, we tested the hypothesis that a tumor suppressor-axis inhibition protein 2 (Axin2) deficiency leads to PAH. METHODS AND RESULTS: We measured right ventricular systolic pressure in Axin2 knockout mice and assessed the expression of Axin2 in patients. We found that Axin2 expression level was decreased in both mice exposed to chronic hypoxia and patients with PAH in remodeled pulmonary arterioles. Axin2 knockout mice showed elevated mean right ventricular systolic pressure and enhanced contraction in response to phenylephrine. An increase in the cross-sectional area of the vessels was occupied by the vessel wall, indicating pulmonary vascular remodeling. Furthermore, knocking down Axin2 with small interfering RNA inhibited apoptosis of pulmonary arterial smooth muscle cells (PASMCs). This inhibition was significantly abolished by ß-catenin inhibitors, indicating that Axin2 through ß-catenin increased vascular wall by inhibiting the apoptosis of PASMCs. Importantly, overexpression of Axin2 attenuates the development of hypoxia-induced PAH in mice. CONCLUSION: Taken together, our study, for the first time, established that Axin2 plays a key role in the progression of PAH. We identified Axin2 as a novel mediator of pulmonary vasoconstriction and PASMC growth in hypoxia-mediated PAH. Our results suggest that downregulation of Axin2 in the pulmonary vasculature may be an underlying mechanism in the development of hypoxia-induced PAH.

[Efficacy of compound xuanju capsule on type-III prostatitis-related sexual dysfunction].

OBJECTIVE: To investigate the value of Compound Xuanju Capsule in the treatment of type-III prostatitis-related sexual dysfunction. METHODS: We randomly divided 90 type-III prostatitis patients with sexual dysfunction diagnosed by NIH clinical criteria into an experiment group and a control group to be treated with Compound Xuanju Capsule and antibiotics, respectively. We analyzed the therapeutic results based on the scores on chronic prostatitis symptom index (CPSI), prostatitis-related sexual function index (PSFI ) and self-rating anxiety scale (SAS), and compared them between the two groups and with the baseline data. RESULTS: The degree of prostatitis-related sexual dysfunction was not correlated with that of prostatitis symptoms. Prostatitis symptoms and sexual function were significantly improved in the experiment group than in the control (P < 0.05), and the SAS score was markedly lower in the former than in the latter (P < 0.05). CONCLUSION: Compound Xuanju Capsule can not only alleviate the symptoms of type-III prostatitis, but also improve its related sexual dysfunction and anxiety.

[Effects of Ginkgo biloba extract on the expression of eNOS and the release of NO in mesenteric arterioles of senile rats].

OBJECTIVE: To investigate the effects of Ginkgo biloba extract (GBE) on the expression of endothelial nitric oxide synthase (eNOS) and the release of nitric oxide (NO) in mesenteric arterioles of aging rats. METHODS: (1) Cytologic experiment:human umbilical vein endothelial cells (HUVEC) were randomly divided into 3 groups: control group, NG-nitro-L-arginine methyl ester (L-NAME) group and GBE group. L-NAME group: 100 µmol/L L-NAME was added into HUNEC for a 48-hour incubation. GBE group: After HUVEC was exposed to 100 µmol/L L-NAME for 24 hours, 20 g/L GBE was added for another 24-hour co-incubation. Then the expression of eNOS protein was observed in each group. (2) Animal experiment: Thirty-two 24-month-old male SD rats were randomly divided into normal control group (n = 8) and GBE group (n = 24). The GBE group was further divided into 3 groups receiving an orally dosed GBE for 3, 5, 7 days respectively. Afterward the diameter of first-order mesenteric arteriole was measured under the pressures of 20, 40, 60, 80, 100, 120, 140 mm Hg (1 mm Hg = 0.133 kPa) and the elasticity of blood vessels calculated. The release of NO, the expression of eNOS protein and its mRNA in mesenteric arterioles stimulated by the same shear stress (15 dyn/cm(2)) were evaluated respectively. RESULTS: (1) Cytological studies indicated that the expression of eNOS protein of the L-NAME group was significantly lower than those of the control and GBE groups (0.57 ± 0.06 vs 0.96 ± 0.05, 0.81 ± 0.09, both P < 0.01). (2) After the dosing of GBE for 3, 5, 7 days, the release of NO was significantly higher than that of the control group [(8.01 ± 0.24, 12.11 ± 0.78, 14.72 ± 0.70 vs 5.83 ± 0.75) pmol×mm(-2)×min(-1), all P < 0.05]; the expressions of eNOS protein were significantly higher than those of the control group (0.59 ± 0.20, 0.86 ± 0.02, 1.09 ± 0.13 vs 0.41 ± 0.16, all P < 0.05). And GBE was highest at Day 7; the expression levels of eNOS mRNA were significantly higher than those of the control group (0.79 ± 0.04, 0.85 ± 0.07, 0.99 ± 0.03 vs 0.58 ± 0.05, all P < 0.05). And GBE was also highest at Day 7. CONCLUSION: GBE can improve vascular flexibility through increasing the expression of eNOS and the release of NO, protecting the functions of blood vessels.