Red Rice Seed Coat Targeting SPHK2 Ameliorated Alcoholic Liver Disease via Restored Intestinal Barrier and Improved Gut Microbiota in Mice.

Alcoholic liver disease (ALD), leading to the most common chronic liver diseases, is increasingly emerging as a global health problem, which is intensifying the need to develop novel treatments. Herein, our work aimed to estimate the therapeutic efficacy of red rice (Oryza sativa L.) seed coat on ALD and further uncover the underlying mechanisms. Red rice seed coat extract (RRA) was obtained with citric acid-ethanol and analyzed via a widely targeted components approach. The potential targets of RRA to ALD were predicted by bioinformatics analysis. Drunken behavior, histopathological examination, liver function, gut microbiota composition and intestinal barrier integrity were used to assess the effects of RRA (RRAH, 600 mg/kg·body weight; RRAL, 200 mg/kg·body weight) on ALD. Oxidative stress, inflammation, apoptosis associated factors and signaling pathways were measured by corresponding kits, Western blot and immunofluorescence staining. In ALD model mice, RRA treatment increased sphingosine kinase 2 (SPHK2) and sphingosine-1-phosphate (S1P) levels, improved gut microbiota composition, restored intestinal barrier, decreased lipopolysaccharide (LPS) levels in plasma and the liver, cut down Toll-like receptor 4 (TLR4)/Nuclear factor kappa B (NF-κB) pathways, alleviated liver pathological injury and oxidative stress, attenuated inflammation and apoptosis and enhanced liver function. To sum up, RRA targeting SPHK2 can ameliorate ALD by repairing intestinal barrier damage and reducing liver LPS level via the TLR4/NF-κB pathway and intestinal microbiota, revealing that red rice seed coat holds potential as a functional food for the prevention and treatment of ALD.

Oryza sativa L. Indica Seed Coat Ameliorated Concanavalin A-Induced Acute Hepatitis in Mice via MDM2/p53 and PKCα/MAPK1 Signaling Pathways.

Acute hepatitis (AH) is a common liver disease with an increasing number of patients each year, requiring the development of new treatments. Hence, our work aimed to evaluate the therapeutic effect of Oryza sativa L. indica (purple rice) seed coat on concanavalin A (ConA)-induced AH and further reveal its potential mechanisms. Purple rice seed coat extract (PRE) was extracted with hydrochloric acid ethanol and analyzed through a widely targeted components method. We evaluated the effects of PRE on AH through histopathological examination, liver function, gut microbiota composition, and the intestinal barrier. The potential targets of PRE on AH were predicted by bioinformatics. Western blotting, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) staining, and corresponding kits were used to investigate PRE effects on predicting targets and associated signaling pathways in AH mice. In AH model mice, PRE treatment increased transformed mouse 3T3 cell double minute 2 (MDM2) expression to inhibit apoptosis; it also markedly downregulated protein kinase C alpha (PKCα), prostaglandin-endoperoxide synthase 1 (PTGS1), and mitogen-activated protein kinase 1 (MAPK1) activity to alleviate inflammation. Thus, PRE treatment also recovered the intestinal barrier, decreased the lipopolysaccharide (LPS) levels of plasma and the liver, enhanced liver function, and improved the composition of intestinal microbiota. In general, PRE targeting MDM2, PKCα, MAPK1, and PTGS1 ameliorated ConA-induced AH by attenuating inflammation and apoptosis, restoring the intestinal barrier, enhancing the liver function, and improving the gut microbiota, which revealed that the purple rice seed coat might hold possibilities as a therapeutic option for AH.

miR-30a-3p Regulates Autophagy in the Involution of Mice Mammary Glands.

The mammary gland undergoes intensive remodeling during the lactation cycle, and the involution process of mammary gland contains extensive epithelial cells involved in the process of autophagy. Our studies of mice mammary glands suggest that miR-30a-3p expression was low during involution compared with its high expression in the mammary glands of lactating mice. Then, we revealed that miR-30a-3p negatively regulated autophagy by autophagy related 12 (Atg12) in mouse mammary gland epithelial cells (MMECs). Restoring ATG12, knocking down autophagy related 5 (Atg5), starvation, and Rapamycin were used to further confirm this conclusion. Overexpression of miR-30a-3p inhibited autophagy and altered mammary structure in the involution of the mammary glands of mice, which was indicative of alteration in mammary remodeling. Taken together, these results elucidated the molecular mechanisms of miR-30a-3p as a key induction mediator of autophagy by targeting Atg12 within the transition period between lactation and involution in mammary glands.

Elsholtzia bodinieri Vaniot ameliorated acute lung injury in mice by regulating pyroptosis, inflammation, oxidative stress and macrophage polarization.

ETHNOPHARMACOLOGICAL RELEVANCE: Elsholtzia bodinieri Vaniot, perennial herbs, a traditional Yunnan Chinese herbal medicine. Its whole herb can be used as commonly used herbs to cure fever, headache, inflammation, indigestion etc., and its tender tip can also be used as tea in Yunnan of China. However, the protective mechanism of Elsholtzia bodinieri Vaniot on acute lung injury (ALI) still needs to be explored. AIM OF STUDY: ALI is characterized by acute respiratory inflammation, which remains a significant source of morbidity and mortality. The current study with the aim of determining the therapeutic the efficacy of E. bodinieri Vaniot on lipopolysaccharide-induced ALI, moreover uncovered the underlying gene-regulated framework, so E. bodinieri Vaniot might serve as functional food for adjuvant therapy or therapeutic agent. MATERIALS AND METHODS: These potential pharmacological targets of E. bodinieri Vaniot against ALI were analyzed by multiple bioinformatics databases. E. bodinieri Vaniot methanol extract (EBE) was obtained by ultrasonic-assisted extraction method, and detected by UHPLC-ESI-HRMS/MS. These pyroptosis, inflammation and oxidative stress associated factors were measured using ELISA assay, western blotting, and histopathological examination to assess the effects of EBE. EcoTyper and immunofluorescence staining were employed to estimate macrophage polarization states in ALI lungs tissue. RESULTS: In ALI lung tissues, EBE treatment could increase B cell leukemia/lymphoma 2 (BCL2) to inhibit pyroptosis, downregulate prostaglandin-endoperoxide synthase 2 (PTGS2) to attenuate inflammation, upregulating NAD(P)H dehydrogenase, quinone 1 (NQO1) to alleviate oxidative stress and induce macrophage polarization toward the M2 phenotype. CONCLUSION: E. bodinieri Vaniot ameliorated ALI thought regulating pyroptosis, inflammation, oxidative stress and macrophage polarization, as well as could be a promising source for therapeutic agent.

Elsholtzia bodinieri Vaniot Ameliorated Acute Lung Injury by NQO1, BCL2 and PTGS2 In Silico and In Vitro Analyses.

Acute lung injury (ALI) is a clinical respiratory disease caused by various factors, which lacks effective pharmacotherapy to reduce the mortality rate. Elsholtzia bodinieri Vaniot is an annual herbaceous plant used as a traditional herbal tea and folk medicine. Here we used bioinformatic databases and software to explore and analyze the potential key genes in ALI regulated by E. bodinieri Vaniot, including B cell leukemia/lymphoma 2 (Bcl2), prostaglandin-endoperoxide synthase 2 (Ptgs2) and NAD(P)H dehydrogenase, quinone 1 (Nqo1). In an inflammatory cells model, we verified bioinformatics results, and further mechanistic analysis showed that methanol extract of E. bodinieri Vaniot (EBE) could alleviate oxidative stress by upregulating the expression of NQO1, suppress pyroptosis by upregulating the expression of BCL2, and attenuate inflammation by downregulating the expression of PTGS2. In sum, our results demonstrated that EBE treatment could alleviate oxidative stress, suppress pyroptosis and attenuate inflammation by regulating NQO1, BCL2 and PTGS2 in a cells model, and E. bodinieri Vaniot might be a promising source for functional food or as a therapeutic agent.

Knockdown of PEBP4 inhibits human glioma cell growth and invasive potential via ERK1/2 signaling pathway.

Phosphatidylethanolamine (PE)-binding protein 4 (PEBP4) is an antiapoptotic protein that is aberrantly expressed in various malignancies. We previously demonstrated that PEBP4 expression is dramatically induced in human gliomas and positively correlated with tumor grade and patient survival. However, the function of PEBP4 in human glioma development and underlying mechanisms remain largely unknown. By stable lentiviral vector-mediated silencing of PEBP4, we examined the effects of PEBP4 knockdown on the growth, apoptosis, and invasion of U251 and U373 human glioma cell lines using MTT, Transwell, colony formation, and flow cytometric assays. We examined the in vivo role of PEBP4 in tumor growth by inoculation of BALB/c nu/nu male mice with PEBP4-deficient U251 and U373 cells. The expression of cell cycle- and apoptosis-related proteins was analyzed by Western blotting and immunostaining. Knockdown of PEBP4 significantly reduced the proliferation and invasion of human glioma cells while inducing cell apoptosis by altering the expression of cell cycle- and apoptosis-related proteins. Mechanistically, PEBP4 knockdown led to activation of the extracellular signal-regulated kinases 1/2 (ERK1/2) pathway, an effect that could be reversed by U0126, a selective inhibitor of MEK1/2 (upstream of ERK1/2), suggesting involvement of ERK1/2 signaling in the regulation of glioma development and progression by PEBP4. We identified PEBP4 as a novel regulator mediating human glioma cell proliferation, invasion, and apoptosis as well as tumor formation and growth. Therefore, PEBP4 may be a potential therapeutic target in human glioma treatment.

[Effectiveness of bone-anterior cruciate ligament-bone allograft in reconstruction of anterior cruciate ligament under arthroscope].

OBJECTIVE: The anterior cruciate ligament (ACL) is the important stable structure of the knee. To evaluate the method and outcome of bone-ACL-bone (B-ACL-B) allograft under arthroscope in reconstruction of ACL. METHODS: Between October 2007 and February 2010, arthroscopic ACL reconstruction with deep-freezing B-ACL-B allograft was performed on 22 patients with ACL ruptures. There were 15 males and 7 females with an average age of 27.6 years (range, 19-55 years). The causes of ACL rupture were sport trauma in 12 cases, falling injury in 1 case, heavy crush in 2 cases, and traffic accident in 7 cases. The locations were the left knee in 14 cases and the right knee in 8 cases. The disease duration was 7 days to 12 months (median, 65 days). Nineteen patients showed the positive results of anterior drawer test and pivot shift test, and 21 patients showed the positive results of Lachman test. According to International Knee Documentation Committee (IKDC) criteria, there were 5 abnormal and 17 severely abnormal. The subjective IKDC score was 49.6 +/- 6.9. The Lysholm score as 48.5 +/- 5.3. The Tegner scale scores were 6.8 +/- 1.2 before injury and 2.1 +/- 0.5 before operation. The MRI showed the ACL injuries in 18 of 20 patients. RESULTS: The mean operative time was 75 minutes (range, 65-85 minutes); the mean blood loss was 110 mL (range, 80-150 mL). All incisions healed by first intention. No immunologic rejection and deep vein thrombosis of lower limbs occurred. All patients were followed up 7-34 months (mean, 18 months). At last follow-up, the flexion of the knee ranged from 125 to 135 degrees (mean, 130.5 degrees). Two patients showed the positive results of anterior drawer test, 1 patient showed the positive result of pivot shift test, and 3 patients showed the positive results of Lachman test. According to the IKDC criteria, 10 patients rated as normal, 11 patients as nearly normal, 1 patient as abnormal. The subjective IKDC score was 90.0 +/- 5.8, showing significant difference when compared with preoperative one (t = 4.653, P = 0.021). The Lysholm score was 91.6 +/- 7.1, showing significant difference when compared with preoperative one (t = 4.231, P = 0.028). The Tegner scale score was 6.1 +/- 1.5, showing no significant difference when compared with one before injury (t = 1.321, P = 0.070) and showing significant difference when compared preoperative one (t = 3.815, P = 0.033). The arthroscopic examination showed no rupture of grafts in 19 patients, 17 grafts showed normal tension, and 2 showed slight relaxation at 6 months after operation. CONCLUSION: Reconstruction of the ACL with B-ACL-B allograft under arthroscope is a safe and effective method, which can anatomically reconstruct ACL and obtain a good recovery of the knee function after operation.

[Treatment of femoral unstable intertrochanteric fractures in the elderly with uncemented long stem bipolar prosthetic prosthesis].

OBJECTIVE: To evaluate the effect of cemented long stem bipolar prosthetic prosthesis in the management of unstable intertrochanteric fractures in the elderly. METHODS: A retrospective analysis of 23 elderly cases of uncemented long-stem prosthetic replacement for unstable intertrochanteric fractures was conducted. Of the 23 elderly patients, 6 had type IIA fracture by Evans-Jenson classification, 11 had type IIB fractures, and 6 had type III fractures. RESULTS: One patient died due to severe respiratory failure during the follow-up period. The patients were followed up for 6 to 10 months (mean 15 months), and no dislocation, infection, loosening or sinking of the prosthesis was found in the followed-up cases with a Harris score reaching 84.8. CONCLUSION: Cemented long-stem bipolar prosthetic prosthesis replacement is effective for management of unstable intertrochanteric fractures in the elderly.