Rotundic acid improves nonalcoholic steatohepatitis in mice by regulating glycolysis and the TLR4/AP1 signaling pathway.

BACKGROUND: Steatosis and inflammation are the hallmarks of nonalcoholic steatohepatitis (NASH). Rotundic acid (RA) is among the key triterpenes of Ilicis Rotundae Cortex and has exhibited multipronged effects in terms of lowering the lipid content and alleviating inflammation. The study objective is to systematically evaluate the potential mechanisms through which RA affects the development and progression of NASH. METHODS: Transcriptomic and proteomic analyses of primary hepatocytes isolated from the control, high-fat diet-induced NASH, and RA treatment groups were performed through Gene Ontology analysis and pathway enrichment. Hub genes were identified through network analysis. Integrative analysis revealed key RA-regulated pathways, which were verified by gene and protein expression studies and cell assays. RESULTS: Hub genes were identified and enriched in the Toll-like receptor 4 (TLR4)/activator protein-1 (AP1) signaling pathway and glycolysis pathway. RA reversed glycolysis and attenuated the TLR4/AP1 pathway, thereby reducing lipid accumulation and inflammation. Additionally, lactate release in L-02 cells increased with NaAsO2-treated and significantly decreased with RA treatment, thus revealing that RA had a major impact on glycolysis. CONCLUSIONS: RA is effective in lowering the lipid content and reducing inflammation in mice with NASH by ameliorating glycolysis and TLR4/AP1 pathways, which contributes to the existing knowledge and potentially sheds light on the development of therapeutic interventions for patients with NASH.

H19 is involved in the regulation of inflammatory responses in acute gouty arthritis by targeting miR-2-3p.

A great number of studies have confirmed that long noncoding RNA (lncRNA) are involved in the regulation of inflammatory response in acute gouty arthritis (AGA). This paper aimed to survey the regulatory mechanism of H19 on AGA. The expression of serum H19 in all subjects was examined by qRT-PCR. The ROC curve was used to estimate the diagnostic value of H19 for AGA. THP-1 cells were induced by MSU to establish in vitro AGA cell model. The concentrations of cytokines such as IL-1ß, IL-8, and TNF-α were tested by ELISA. Luciferase reporter gene analysis was used to verify the interaction between H19 and the 3'-UTR of miR-22-3p. Expressions of serum H19 in AGA patients were significantly higher than that in controls. The ROC curve indicated the potential of H19 as a diagnostic marker for AGA. Cell experiments revealed that the downregulation of H19 significantly inhibited the expressions of IL-1ß, IL-8, and TNF-α. The luciferase reporter gene assay manifested that miR-22-3p is the target gene of H19. And knockdown of miR-22-3p overturned the downregulation of inflammatory factors caused by H19 inhibition. H19 aggravated MSU-induced THP-1 inflammation by negatively targeting miR-22-3p, suggesting a new regulatory mechanism and potential therapeutic target for AGA.

Long non-coding RNA SND1-IT1 accelerates cell proliferation, invasion and migration via regulating miR-132-3p/SMAD2 axis in retinoblastoma.

Long noncoding RNAs (lncRNAs) have been identified as prognostic biomarkers and functional regulators in human tumors. In our study, we aim to investigate the roles of lncRNA SND1-IT1 (SND1-IT1) in retinoblastoma (RB). We observed that SND1-IT1 was highly expressed in both RB specimens and cells, and associated with poorer prognosis of RB patients. Functional investigation revealed that downregulation of SND1-IT1 suppressed RB cell proliferation, migration and invasion in vitro and restrained RB tumorigenesis in vivo. MiR-132-3p was predicted to interact with SND1-IT1. RT-qPCR and dual-luciferase reporter assays verified the regulation of miR-132-3p by SND1-IT1 in RB cells. In addition, SND1-IT1 enhanced the expression of SMAD2 by sponging miR-132-3p. Rescue experiments revealed that knockdown of miR-132-3p reversed the inhibiting effects of miR-132-3p knockdown on RB cells. Overall, SND1-IT1 can promote the progression of RB cells through miR-132-3p/SMAD2 axis, suggesting that l SND1-IT1 might be a novel biomarker and potential target for RB.

Rotundic acid ameliorates non-alcoholic steatohepatitis via SREBP-1c/ SCD1 signaling pathway and modulating gut microbiota.

Non-alcoholic steatohepatitis (NASH) is a devastating form of non-alcoholic fatty liver disease (NAFLD) with distinguished hallmarks of steatosis and inflammation. Rotundic acid (RA) is a natural pentacyclic triterpene compound extracted from the bank of Ilex rotunda Thunb with a wide range of biological activities. The aim of the study is to evaluate the pharmacological effect and action mechanism of RA on NASH in vitro and in vivo. RA has weak lipid lowering ability in rat primary hepatocytes, significantly decreases serum LDL level, hepatic TG and TC levels and lipid droplets, reduces NAS compared with the NASH group, and alleviates hepatic inflammation. RA also enhances the recovery of intestinal bacterial community and intestinal-derived short-chain fatty acid caused by high food diet (HFD). Further investigation shows that RA protects against HFD-induced NASH via downregulating the expression of SREBP-1c/SCD1 signaling pathway and improving gut microbiota. These findings imply that RA might be helpful for the alleviation of NASH.

Inhibition of Nav1.7 channel by a novel blocker QLS-81 for alleviation of neuropathic pain.

Voltage-gated sodium channel Nav1.7 robustly expressed in peripheral nociceptive neurons has been considered as a therapeutic target for chronic pain, but there is no selective Nav1.7 inhibitor available for therapy of chronic pain. Ralfinamide has shown anti-nociceptive activity in animal models of inflammatory and neuropathic pain and is currently under phase III clinical trial for neuropathic pain. Based on ralfinamide, a novel small molecule (S)-2-((3-(4-((2-fluorobenzyl) oxy) phenyl) propyl) amino) propanamide (QLS-81) was synthesized. Here, we report the electrophysiological and pharmacodynamic characterization of QLS-81 as a Nav1.7 channel inhibitor with promising anti-nociceptive activity. In whole-cell recordings of HEK293 cells stably expressing Nav1.7, QLS-81 (IC50 at 3.5 ± 1.5 µM) was ten-fold more potent than its parent compound ralfinamide (37.1 ± 2.9 µM) in inhibiting Nav1.7 current. QLS-81 inhibition on Nav1.7 current was use-dependent. Application of QLS-81 (10 µM) caused a hyperpolarizing shift of the fast and slow inactivation of Nav1.7 channel about 7.9 mV and 26.6 mV, respectively, and also slowed down the channel fast and slow inactivation recovery. In dissociated mouse DRG neurons, QLS-81 (10 µM) inhibited native Nav current and suppressed depolarizing current pulse-elicited neuronal firing. Administration of QLS-81 (2, 5, 10 mg· kg-1· d-1, i.p.) in mice for 10 days dose-dependently alleviated spinal nerve injury-induced neuropathic pain and formalin-induced inflammatory pain. In addition, QLS-81 (10 µM) did not significantly affect ECG in guinea pig heart ex vivo; and administration of QLS-81 (10, 20 mg/kg, i.p.) in mice had no significant effect on spontaneous locomotor activity. Taken together, our results demonstrate that QLS-81, as a novel Nav1.7 inhibitor, is efficacious on chronic pain in mice, and it may hold developmental potential for pain therapy.

Prognostic factors affecting the short-term efficacy of non-surgical treatment of chronic periodontitis: a multilevel modeling analysis.

BACKGROUND: This study is aimed to analyze the prognostic factors affecting the short-term efficacy of non-surgical treatment of patients in periodontitis from stage II to stage IV by the multilevel modeling analysis. MATERIALS AND METHODS: A total of 58 patients with chronic periodontitis were included in this study. Patients were clinically explored before and 3 months after the treatment and the difference in probing depth was determined [Reduction of probing depth (Δ PD) = baseline PD - finial probing depth (FPD)] which is considered as the therapeutic evaluation. Three different levels were analyzed: patients, teeth and sites to construct a multi-layer linear model. RESULTS: Probing depth (PD) improved significantly compared with that before treatment (p < 0.05), in which FPD was (3.90 ± 1.39) mm, and the ΔPD was (1.79 ± 0.97) mm. Compared with the mesial sites and distal sites of the multi-rooted teeth, the number of PD ≥ 5 mm or PD < 5 mm after the treatment was significantly different (P < 0.05), and the proportion of PD < 5 mm was higher in mesial sites. The null model showed that Δ PD varied greatly between groups at various levels (P < 0.001), with prediction variable of site level, tooth level, and patient level accounted for 66%, 18%, and 16% of the overall difference, respectively. The complete model showed that the Δ PD of smokers was significantly lower than that of non-smokers (P < 0.001). The Δ PD of the mesial and distal sites was larger than that of the buccolingual central site (P < 0.001). The Δ PD of single-rooted teeth was larger than that of multi-rooted teeth (P < 0.001). The baseline PD, tooth mobility (TM), bleeding index (BI), clinical attachment loss (CAL) were significantly negatively correlated with Δ PD (P < 0.001). CONCLUSIONS: Patients with periodontitis from stage II to stage IV, who were non-smoking, have good compliance, good awareness of oral health, and low percentage sites with PD ≥ 5 mm at baseline, single-rooted teeth with hypomobility, less clinical attachment loss and lower bleeding index and sites of mesial or distal can obtain an ideal short-term efficacy of non-surgical treatment.

Two new cephalochromin derivative from the Alternaria sp. ZG22.

Two new cephalochromin derivatives, prenylcephalochromin A (1), prenylcephalochromin B (2), along with cephalochromin (3) were isolated from the Alternaria sp. ZG22 obtained from a Dasymaschalon rostratum collected from the Hainan. The structures of two new compounds were elucidated by comprehensive spectroscopic methods. Compounds 1-3 showed α-glucosidase inhibitory activity.

Structural variation of the complete chloroplast genome and plastid phylogenomics of the genus Asteropyrum (Ranunculaceae).

Two complete chloroplast genome sequences of Asteropyrum, as well as those of 25 other species from Ranunculaceae, were assembled using both Illumina and Sanger sequencing methods to address the structural variation of the cp genome and the controversial systematic position of the genus. Synteny and plastome structure were compared across the family. The cp genomes of the only two subspecies of Asteropyrum were found to be differentiated with marked sequence variation and different inverted repeat-single copy (IR-SC) borders. The plastomes of both subspecies contains 112 genes. However, the IR region of subspecies peltatum carries 27 genes, whereas that of subspecies cavaleriei has only 25 genes. Gene inversions, transpositions, and IR expansion-contraction were very commonly detected in Ranunculaceae. The plastome of Asteropyrum has the longest IR regions in the family, but has no gene inversions or transpositions. Non-coding regions of the cp genome were not ideal markers for inferring the generic relationships of the family, but they may be applied to interpret species relationship within the genus. Plastid phylogenomic analysis using complete cp genome with Bayesian method and partitioned modeling obtained a fully resolved phylogenetic framework for Ranunculaceae. Asteropyrum was detected to be sister to Caltha, and diverged early from subfamily Ranunculoideae.

The synergistic effect of cobalt on a Pd/Co catalyzed Suzuki-Miyaura cross-coupling in water.

This work introduces the new trimetallic complex CoPd2(HBPDC)2Cl4·(H2O)4(H2BPDC = 2,2'-bipyridine-4,4'-dicarboxylic acid) as a highly efficient and more cost-effective catalyst for a Suzuki-Miyaura reaction proceeding in water, without additives and under aerobic conditions. Catalytic studies revealed a synergistic Co-Pd cooperativity, fostered by ligation through H2BPDC, and accounting for the superior performance of the heterobimetallic complex vs. its Co-free counterpart.

[IDH1 Gene Mutation and Its Clinical Significance in patients with Acute Myeloid Leukemia].

OBJECTIVE: To evaluate the incidence rate of IDH1 in acute myeloid leukemia and analyze its effect on clinical characteristics and prognosis. METHODS: Mononuclear cells in bone marrow samples were collected from 192 adult patients with newly diagnosed AML. Polymerase chain reaction (PCR) and direct sequencing were used to amplify exon 4 of IDH1 gene, the gene sequencing was used to analyze the gene mutations, at same time, the detection of NPM1, FLT3-TKD, FLT3-ITD, C-KIT, CEPBA, TET2 and JAK2V617F and MLL mutations were carried out, the follow-up was used to determine its therapeutic efficacy and outcomes of patients. The clinical and laboratory data of these cases were collected, and their clinical characteristics and prognosis were then analyzed. RESULTS: Among the 192 AML patients, 13 cases were detected with IDH1 gene mutation, the mutation rate was 6.77% [95% CI (5.70%-13.38%)]. The sequencing chart of IDH1 gene showed double peaks, the mutations were heterozygous, out of them c.G395A (p.R132H) was found in 8 cases, c.C394T was found in 4 cases (p.R132C), c.C394A (p.R132S) was found in 1 cases, R132H and R132C are common, 13 cases showed missense mutation. The median age in mutation group was 52 years old, the median age in unnutration group was 40 years, there was significant difference between them (P = 0.010). Mutation rate of IDH1 gene in M1 and M2 was significantly higher than that in other FAB subtypes. There were no significant difference in sex, newly diagnosed peripheral white blood cell count, hemoglobin, platelet count, peripheral blood and bone marrow original cell proportion of primitive cells between them. Mutation of IDH1 gene had certain correlation with NPM1 gene mutation, but no correlation with FLT3-TKD, FLT3-ITD, C-KIT, TET2 and JAK2V617F and MLL natations was found. In addition, the IDH1 mutation easily occurred in patients with normal karyotype or in patients with middle prognostic risk karyotype, IDH1 mutation occurred in 11 cases with normal karyotype, the mutation rate was 10.28%, IDH1 mutation were observed in 2 cases with abnormal karyotype, the mutation rate was 3.50%, there was significant difference. In AML patients with middle prognostic risk karyotype. The complete remission (CR) and the 3 year survival (OS) rate of IDH1 mut patients were less than that in IDH1 wt, there was significant difference (P < 0.05). CONCLUSIONS: The IDH1 mutation more easily occurr in older AML patients and mutations effect of IDH1 on clinical characteristics may represent a molecular marker for poor prognosis in AML.

[Ecological compensation standard in Dongting Lake region of returning cropland to lake based on emergy analysis].

The annual emergy and currency value of the main ecological service value of returning cropland to lake in Dongting Lake region from 1999 to 2010 was calculated based on emergy analysis. The calculation method of ecological compensation standard was established by calculating annual total emergy of ecological service function increment since the starting year of returning cropland to lake, and the annual ecological compensation standard and compensation area were analyzed from 1999 to 2010. The results indicated that ecological compensation standard from 1999 to 2010 was 40.31-86.48 yuan x m(-2) with the mean of 57.33 yuan x m(-2). The ecological compensation standard presented an increase trend year by year due to the effect of eco-recovery of returning cropland to lake. The ecological compensation standard in the research area presented a swift and steady growth trend after 2005 mainly due to the intensive economy development of Hunan Province, suggesting the value of natural ecological resources would increase along with the development of society and economy. Appling the emergy analysis to research the ecological compensation standard could reveal the dynamics of annual ecological compensation standard, solve the abutment problem of matter flow, energy flow and economic flow, and overcome the subjective and arbitrary of environment economic methods. The empirical research of ecological compensation standard in Dongting Lake region showed that the emergy analysis was feasible and advanced.

[Comparison of organic component and di-n-butyl phthalate between human milk and cow milk products].

OBJECTIVE: To explore types of organic components and pollution level of di-n-butyl phthalate (DBP) between human milk and cow milk products. METHODS: Forty healthy postpartum women with an average age of (27.44 ± 3.43) years old were selected, and a 5 ml sample of breast milk were collected. Four different brands of fresh cow milk and 1 brand of milk powder were randomly selected in the market. A total of 15 samples were collected with 3 from each brand, and the qualitative analysis of types of organic components and quantitative analysis of DBP were conducted by gas-chromatography and mass-spectrometry (GC/MS) method. RESULTS: A total of 176 different types of organic components were detected in 40 samples of human milk (averaged at (10.58 ± 4.16) types per sample); 37 different types were detected in 12 samples of fresh cow milk (averaged at (8.67 ± 1.61) types per sample); while 31 types of organic components were detected in 3 samples of milk powder (averaged at (12.67 ± 0.58) types per sample). It was obvious that the types of organic components in milk powder were significantly higher than the other two groups (t = 2.09, 4.00, P < 0.05). The most frequent organic component in human milk and cow milk was 9-octadecenoic acid (45.00% (18/40) in human milk; 53.33% (8/15) in cow milk). DBP concentrations were (57.78 ± 35.42) µg/L, (20.76 ± 6.60) µg/L and (0.45 ± 0.05) mg/kg (equal to (66.78 ± 7.60) µg/L) in human milk, fresh cow milk and milk powder, respectively. The DBP concentration in fresh cow milk was significantly lower than those in human milk and milk powder (t = 37.02, 46.02, P < 0.05). CONCLUSION: Both human milk and cow milk contain different types of organic pollutants, some of which have toxic effects on reproduction and human development.

[Selection and anti-cancer effects of siRNAs targeting HMGA2 gene].

High mobility group A2 protein (HMGA2), an architectural factor, is highly expressed in various cancer types including lung cancers. It is a candidate target for cancer therapy. RNAi is an effective gene silencing method with low cost and less time-consuming. It is possible to exploit this technology in therapy. Here, 5 siRNAs targeting Hmga2 gene (HMGA2 siRNA1-5) were designed and synthesized. MTT assay, colony formation assay, transwell assay and flow cytometry were used to evaluate the effects of these siRNAs on lung cancer cell lines (NCI-H446 and A549). Results from cell proliferation, clone formation, migration and apoptosis showed that HMGA2 siRNA1, 3, 5 could affect these aspects for both lung cancer cell lines. Among the five siRNAs, HMGA2 siRNA5 showed the greatest inhibition effects. The inhibition effects of HMGA2 siRNA5 are sequence specific and are not due to the induction of interferon response. Taken together, siRNAs targeting Hmga2 gene are potential candidates for lung cancer gene therapy.

The effect of different acclimation temperatures on the prophenoloxidase system and other defence parameters in Litopenaeus vannamei.

Water temperature changes (higher and lower than 24 degrees C) were shown to have a significant effect on dopamine (DA) concentration, haemocyte count and the proPO system in the white shrimp Litopenaeus vannamei. No significant difference in any of the parameters was observed in the control group. DA concentration in haemolymph in the experimental groups increased to a peak value at 0.5 days; meanwhile serine protease (SP) activity and proteinase inhibitor (PI) activity decreased. Total haemocyte count (THC), differential haemocyte count (DHC) and PO activity were lowest at 1 day. All defence parameters became stable after 1-3 days, while the total haemocyte and large granular cell count stabilized after 6 days. After these stabilized, there was no significant difference in DA concentration and PI activity between the control and experimental groups, as was the case for the THC, DHC, PO and SP activities of shrimp held at higher temperatures. However these latter four parameters in the lower temperature groups were distinctly lower than the control group. alpha(2)-Macroglobulin activity in the experimental groups increased to a peak value after 1 day compared with the control and then stabilized after 6 days when the activity levels in higher temperature groups were higher than the control, while the lower temperature groups had no significant difference from the control. It was therefore concluded that water temperature changes modulated the immune system of L. vannamei.

Dynamic expression of pepsinogen C in gastric cancer, precancerous lesions and Helicobacter pylori associated gastric diseases.

AIM: To investigate the relationship between the expression of pepsinogen C (PGC) and gastric cancer, precancerous diseases, and Helicobacter pylori (H pylori) infection. METHODS: The expression of PGC was determined by immunohistochemistry method in 430 cases of gastric mucosa. H pylori infection was determined by HE staining, PCR and ELISA in 318 specimens. RESULTS: The positive rate of PGC expression in 54 cases of normal gastric mucosa was 100%. The positive rates of PGC expression in superficial gastritis or gastric ulcer or erosion, atrophic gastritis or gastric dysplasia and gastric cancer decreased significantly in sequence (P<0.05; 100%/89.2% vs 14.3%/15.2% vs 2.4%). The over-expression rate of PGC in group of superficial gastritis with H pylori infection was higher than that in group without H pylori infection (P<0.05; chi2= 0.032 28/33 vs 15/25). The positive rate of PGC expression in group of atrophic gastritis with H pylori infection was lower than that in group without H pylori infection (P<0.01; chi2= 0.003 4/61 vs 9/30), and in dysplasia and gastric cancer. CONCLUSION: The level of PGC expression has a close relationship with the degree of malignancy of gastric mucosa and development of gastric lesions. There is a relationship between H pylori infection and expression of antigen PGC in gastric mucosa, the positive rate of PGC expression increases in early stage of gastric lesions with H pylori infection such as gastric inflammation and decreases during the late stage such as precancerous diseases and gastric cancer. PGC-negative cases with H pylori-positive gastric lesions should be given special attention.

[Expression of pepsinogen C in gastric cancer and its precursor].

OBJECTIVE: To investigate the dynamic expression of pepsinogen C (PGC) and its value in detection of precursor and gastric cancer. METHODS: Immunohistochemistry was used to examine the expression of PGC in 424 biopsy specimens of stomach mucosa collected by gastroscopy. RESULTS: The positive rate of PGC expression in 54 cases of normal gastric mucosa was 100% and 2.4% in 124 cases of gastric cancer. The positive rate of PGC expression decreased in the order of superficial gastritis/gastric ulcer or erosion-->atrophic gastritis or gastric dysplasia-->gastric cancer (P < 0.01). CONCLUSION: The expression of PGC is negatively correlated with the degree of malignity of gastric mucosa and development of gastric lesions. PGC has high sensitivity and specificity in diagnosis of precursor of gastric cancer and can be a good indicator in the screening and diagnosis of precursor of gastric cancer and gastric cancer.

Association between pepsinogen C gene polymorphism and genetic predisposition to gastric cancer.

AIM: To identify a molecular marker for gastric cancer, and to investigate the relationship between the polymorphism of pepsinogen C (PGC) gene and the genetic predisposition to gastric cancer. METHODS: A total of 289 cases were involved in this study. 115 cases came from Shenyang area, a low risk area of gastric cancer, including 42 unrelated controls and 73 patients with gastric cancer. 174 cases came from Zhuanghe area, a high-risk area of gastric cancer, including 113 unrelated controls, and 61 cases from gastric cancer kindred families. The polymorphism of PGC gene was detected by polymerase chain reaction (PCR) and the relation between the genetic polymorphism of PGC and gastric cancer was examined. RESULTS: Four alleles, 31 0bp (allele 1), 400 bp (allele 2), 450 bp (allele 3), and 480 bp (allele 4) were detected by PCR. The frequency of allele 1 was higher in patients with gastric cancer than that in controls. Genotypes containing homogenous allele 1 were significantly more frequent in patients with gastric cancer than that in controls (0.33, 0.14, chi(2)=3.86, P<0.05). There was no significant difference between the control group of Zhuanghe and the group of gastric cancer kindred. But the frequency of allele 1 was higher in control group of Zhuanghe area than that in control group of Shenyang area and genotypes containing homogenous allele 1 were significantly more frequent in the control group of Zhuanghe area than those in control group of Shenyang area (0.33, 0.14, chi(2)=4.32, P<0.05). In the group of gastric cancer kindred the frequency of allele 1 was significantly higher than that in control group of Shenyang area (0.5164, 0.3571, chi(2)=4.47, P<0.05). Genotypes containing homogenous allele 1 were significantly more frequent in the group of gastric cancer kindred than those in control group of Shenyang area (0.36, 0.14, chi(2)=4.91, P<0.05). CONCLUSION: These results suggest that there is some relation between pepsinogen C gene polymorphism and gastric cancer, and the person with homogenous allele 1 predisposes to gastric cancer than those with other genotypes. Pepsinogen C gene polymorphism may be used as a genetic marker for a genetic predisposition to gastric cancer. The distribution of pepsinogen C gene polymorphism in Zhuanghe, a high-risk area of gastric cancer, is different from that in Shenyang, a low risk area of gastric cancer.