Quality evaluation of Houttuynia cordata Thunb. by high performance liquid chromatography with photodiode-array detection (HPLC-DAD).

A new, validated method, developed for the simultaneous determination of 16 phenolics (chlorogenic acid, scopoletin, vitexin, rutin, afzelin, isoquercitrin, narirutin, kaempferitrin, quercitrin, quercetin, kaempferol, chrysosplenol D, vitexicarpin, 5-hydroxy-3,3',4',7-tetramethoxy flavonoids, 5-hydroxy-3,4',6,7-tetramethoxy flavonoids and kaempferol-3,7,4'-trimethyl ether) in Houttuynia cordata Thunb. was successfully applied to 35 batches of samples collected from different regions or at different times and their total antioxidant activities (TAAs) were investigated. The aim was to develop a quality control method to simultaneously determine the major active components in H. cordata. The HPLC-DAD method was performed using a reverse-phase C18 column with a gradient elution system (acetonitrile-methanol-water) and simultaneous detection at 345 nm. Linear behaviors of method for all the analytes were observed with linear regression relationship (r(2)>0.999) at the concentration ranges investigated. The recoveries of the 16 phenolics ranged from 98.93% to 101.26%. The samples analyzed were differentiated and classified based on the contents of the 16 characteristic compounds and the TAA using hierarchical clustering analysis (HCA) and principal component analysis (PCA). The results analyzed showed that similar chemical profiles and TAAs were divided into the same group. There was some evidence that active compounds, although they varied significantly, may possess uniform anti-oxidant activities and have potentially synergistic effects.

[Culture in vitro and plant regeneration of Panax japonicus].

OBJECTIVE: To study the condition of culture in vitro and plant regeneration of Panax japonicus. METHODS: Embryos, stems and leaves of P. japonicus were used as explants, effects of different hormones for callus induction and plant regeneration were studied and optimized. RESULTS: The optimal way to obtain sterile explant for seeds was sterilized in 75% ethyl alcohol for 60 s then 0.1% HgCL2 for 12 min; Stems and leaves were sterilized in 75% ethyl alcohol for 15 s then 5% NaClO for 5 min. Used MS as basic medium, the optimal hormones combination for callus induction of embryos, stems and leaves were MS + 1.5 mg/L NAA + 1.5 mg/L 2, 4-D + 0.1 mg/L KT; MS + 1.5 mg/L NAA + 1.0 mg/L 2,4-D + 0.1 mg/L KT; MS + 1.5 mg/L NAA + 1.0 mg/L 2,4-D + 0.2 mg/L KT respectindy under the illumination. But under the darkness,the optimal callus induction hormones combination for embryos leaves were MS + 1.0 mg/L NAA + 1.5 mg/L 2,4-D +0.2 mg/L KT; 1.5 mg/L NAA + 1.5 mg/L 2,4-D + 0.1 mg/L KT; MS + 1.5 mg/L NAA + 1.0 mg/L 2,4-D + 0.1 mg/L KT respectivety. The optimal medium for germination was MS + 3.0 mg/L 6-BA + 1.0 mg/L GA3. The optimal medium for roots generation was MS + 1.0 mg/L 6-BA + 3.0 mg/L IBA. CONCLUSION: We establish the system of culture in vitro and plant regeneration for P. japonicus.

Comprehensive circular RNA profiling reveals the regulatory role of circRNA_0007694 in papillary thyroid carcinoma.

PURPOSE: The present study aimed to identify differentially expressed circRNAs in thyroid cancer and verify their potential functions. METHODS: Next-generation sequencing was used to identify differentially expressed circRNAs between papillary thyroid carcinoma (PTC) tissues and paired pericarcinomatous tissues. Polymerase chain reaction and Sanger sequencing methods successfully identified hsa_circ_0007694. A hsa_circ_0007694 over-expression vector was prepared to determine the effect of this circRNA on proliferation, migration, invasion, apoptosis, and the cell cycle in PTC cells. An in vivo animal assay was conducted by injecting PTC cells into the chests of mice. Further, RNA-seq was performed, followed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, to verify the regulatory mechanism of hsa_circ_0007694. Western blotting was used to verify the genes thought to be involved in the hsa_circ_0007694 regulatory pathways based on KEGG analysis. RESULTS: We identified a circRNA, hsa_circ_0007694 that was down-regulated in PTC tissues compared to pericarcinomatous tissues. Over-expression of hsa_circ_0007694 promoted apoptosis and inhibited proliferation, migration, and invasion in PTC cells in vitro, and decreased tumor growth in vivo. Transcriptome sequence analysis suggested 129 differentially expressed genes between PTC tissue and paired pericarcinomatous tissue. KEGG analysis and western blotting indicated that the PI3K/AKT/mTOR and Wnt signaling networks are most likely to be related to hsa_circ_0007694 in thyroid cancer. CONCLUSION: The circRNA hsa_circ_0007694 is down-regulated in PTC and is therefore a potential therapeutic target.

Knockdown of angiopoietin-like 4 inhibits the development of human gastric cancer.

Human gastric cancer (GC) is the second most common cause of cancer-related deaths worldwide and is one of the most common metastatic cancers. Tumor proliferation, apoptosis, metastasis and invasion are important predictors of the invasiveness of GC and are key factors in cancer-induced death. Angiopoietin-like 4 (ANGPTL4) is a secreted protein that belongs to the angiopoietin (ANGPTL) family and is involved in the regulation of cancer metastasis. However, whether ANGPTL4 plays a role in the progression of GC remain unclear. In the present study, immunoreactivity of ANGPTL4 demonstrated that ANGPTL4 expression was upregulated in GC tissues with the development of GC. The siRNA targeting ANGPTL4 effectively knocked down ANGPTL4 in the SNU­1 and BGC823 cell lines at the mRNA and protein levels. Following ANGPTL4 downregulation, the proliferation and invasion abilities of GC cell lines were suppressed as determined by MTT and Transwell assays, and cell apoptosis level and sensitivity to cisplatin were increased as determined by flow cytometry and MTT assay. In conclusion, these findings suggest that ANGPTL4 may be a new potential therapeutic target for GC.

Corrigendum: (-)-Epigallocatechin-3-Gallate Ameliorates Atherosclerosis and Modulates Hepatic Lipid Metabolic Gene Expression in Apolipoprotein E Knockout Mice: Involvement of TTC39B.

[This corrects the article on p. 195 in vol. 9, PMID: 29593532.].

(-)-Epigallocatechin-3-Gallate Ameliorates Atherosclerosis and Modulates Hepatic Lipid Metabolic Gene Expression in Apolipoprotein E Knockout Mice: Involvement of TTC39B.

Background: Aberrant chronic inflammation and excess accumulation of lipids play a pivotal role in the occurrence and progression of atherosclerosis. (-)-Epigallocatechin-3-gallate (EGCG), the major catechins in green tea, displayed anti-atherosclerotic properties in vivo and in vitro. However, the effects and underlying mechanism of EGCG on atherosclerosis remain unclear. Methods: Male apolipoprotein E-knockout (ApoE-/-) mice (7 weeks old) fed with high-fat diet (HFD) were treated with normal saline or EGCG (40 mg/kg/d, i.g.) for 18 weeks. Atherosclerotic plaque and liver lipid accumulation were measured by Oil Red staining. Plasma lipids and cytokines were detected using commercial kits. The expression of protein and mRNA was analyzed by western blot and quantitative real-time reverse transcription-polymerase chain reaction, respectively. Results: EGCG administration markedly attenuated atherosclerotic plaque formation in HFD-fed ApoE-/- mice, which were accompanied by increased plasma interleukin-10 (IL-10) level and decreased plasma IL-6 and tumor necrosis factor-α (TNF-α) levels. In addition, EGCG modulated high-fat-induced dyslipidemia, evidencing by decreased total cholesterol (TC) and low-density lipoprotein levels and increased high-density lipoprotein level. Meanwhile, EGCG treatment alleviated high-fat-mediated liver lipid accumulation and decreased liver TC and triglyceride. Mechanistically, EGCG significantly modulated high-fat-induced hepatic tetratricopeptide repeat domain protein 39B (TTC39B) expression and its related genes (Lxrß, Abcg5, Abcg8, Abca1, Srebf1, Scd1, Scd2, Fas, Elovl5, Mylip) expression in liver from ApoE-/- mice. Notably, EGCG remarkably induced hepatic liver X receptor α (LXRα) and LXRß expression and inhibited both precursor and mature sterol regulatory element binding transcription factor-1 (SREBP-1) expression. Conclusion: Taken together, our data for the first time suggested that TTC39B was involved in EGCG-mediated anti-atherosclerotic effects through modulation of LXR/SREBP-1 pathway.

Impact of Resistance Training in Subjects With COPD: A Systematic Review and Meta-Analysis.

BACKGROUND: The goal of this study was to evaluate the effects of resistance training on subjects with COPD. METHODS: We performed a systematic search in MEDLINE, PubMed, Embase, CINAHL, Elsevier ScienceDirect, EBM Reviews, Cochrane Central Register of Controlled Trials, and ClinicalTrials.gov and also of leading respiratory journals for randomized controlled trials on COPD treatment for ≥ 4 weeks with resistance training compared with non-exercise control or with combined resistance and endurance training compared with endurance training alone. Data from these studies were pooled to calculate odds ratio and weighted mean differences (WMDs) with 95% CI. RESULTS: Eighteen trials with 750 subjects with advanced COPD met the inclusion criteria. There were 2 primary and 5 secondary outcomes. Compared with non-exercise control, resistance training led to significant improvements in the dyspnea domain of the Chronic Respiratory Disease Questionnaire (WMD of 0.59, 95% CI 0.26-0.93, I2 = 0%, P < .001), skeletal muscle strength, and percent-of-predicted FEV1 (WMD of 6.88%, 95% CI 0.41-13.35%, I2 = 0%, P = .04). The combination of resistance and endurance training significantly improved the St George Respiratory Questionnaire total score (WMD of -7.44, 95% CI -12.62 to -2.25, I2 = 0%, P = .005), each domain score, and skeletal muscle strength. There were no significant differences in 6-min walk distance, 6-min pegboard and ring test, maximum exercise work load, and maximum oxygen consumption between the 2 groups. There were no reports of adverse events related to resistance-training intervention. CONCLUSIONS: Resistance training can be successfully performed alone or in conjunction with endurance training without increased adverse events during pulmonary rehabilitation in COPD.

Tiotropium versus placebo for inadequately controlled asthma: a meta-analysis.

OBJECTIVE: This meta-analysis was performed to evaluate the efficacy and safety of the addition of tiotropium to standard treatment regimens for inadequately controlled asthma. METHODS: A systematic search was made of PubMed, EMBASE, MEDLINE, and CENTRAL databases, and ClinicalTrials.gov, and a hand search of leading respiratory journals. Randomized, double-blind clinical trials on the treatment of inadequately controlled asthma for ≥ 4 weeks with the addition of tiotropium, compared with placebo, were reviewed. Studies were pooled to odds ratio (OR) and weighted mean differences (WMDs), with 95% CI. RESULTS: Six trials met the inclusion criteria. The addition of tiotropium, compared with placebo, significantly improved all spirometric indices, including morning and evening peak expiratory flow (WMD 20.59 L/min, 95% CI 15.36-25.81 L/min, P < .001; and WMD 24.95 L/min, 95% CI 19.22-30.69 L/min, P < .001, respectively), trough and peak FEV1 (WMD 0.13 L, 95% CI 0.09-0.18 L, P < .001; and WMD 0.10 L, 95% CI 0.06-0.14 L, P < .001, respectively), the area under the curve of the first 3 h of FEV1 (WMD 0.13 L, 95% CI 0.08-0.18 L, P < .001), trough and peak FVC (WMD 0.1 L, 95% CI 0.05-0.15 L, P < .001; and WMD 0.08 L, 95% CI 0.04-0.13 L, P < .001, respectively), the area under the curve of the first 3 h of FVC (WMD 0.11 L, 95% CI 0.06-0.15 L, P < .001). The mean change in the 7-point Asthma Control Questionnaire score (WMD -0.12, 95% CI -0.21 to -0.03, P = .01) was markedly lower in tiotropium group, but not clinically important. There were no significant differences in Asthma Quality of Life Questionnaire score (WMD 0.09, 95% CI -0.01 to 0.20, P = .09), night awakenings (WMD 0.00, 95% CI -0.05 to 0.05, P = .99) or rescue medication use (WMD -0.18, 95% CI -0.36 to 0.00, P = .06). No significant increase was noticed in adverse events in the tiotropium group (OR 0.80, 95% CI 0.62-1.03, P = .08). CONCLUSIONS: The addition of tiotropium to standard treatment regimens has significantly improved lung function without increasing adverse events in patients with inadequately controlled asthma. Long-term trials are required to assess the effects of the addition of tiotropium on asthma exacerbations and mortality.

Regulation of skeletal muscle differentiation in fibroblasts by exogenous MyoD gene in vitro and in vivo.

MyoD of the myogenic regulatory factors (MRFs) family regulates the skeletal muscle differentiation program. In this study, stably transfected NIH3T3-derived cell lines were established, in which exogenous MyoD was expressed at high levels. Transcriptional activation of endogenous muscle regulatory gene and induction towards the skeletal muscle lineages were observed with phase-contrast microscopy when continuously cultured in vitro. Moreover, to determine their ability of myogenic formation in vivo, the transfected cells were implanted in nude mice subcutaneously for up to 10 weeks. The morphological characterization of inductive cells was observed using transmission electron microscope and histological staining. Myogenesis of fibroblasts incubated in the medium was activated by overexpression of MyoD, and the cells were accumulated and fused into multinucleated myotubes. Correlatively, RT-PCR and immunohistochemistry confirmed the increased expression of characteristic downstream molecule myogenin and mysion heavy chains during myogenic differentiation. Ecoptic myogenesis was found and remained stable phenotype when the transfected cells were seeded in vivo. Our results suggest that MyoD can be considered to be a determining factor of myogenic lineages, and it may play an important role in the cell therapy and cell-mediated gene therapy of the skeletal muscle.

Temporal expression of estrogen receptor alpha in rat bone marrow mesenchymal stem cells.

Estrogen responsiveness of bone formation is mediated by the estrogen receptor alpha (ERalpha) in osteoblastic lineage. As osteoblasts arise from the multipotent bone marrow stromal (mesenchymal) cells, this study was undertaken to observe the ERalpha in primary female adult rat bone marrow mesenchymal stem cells (BMSCs). The ERalpha was localized using immunocytochemical analysis in identified primary BMSCs. Then, using real-time PCR analysis, we measured the expression of ERalpha messenger RNA (mRNA) in BMSCs. ERalpha transcripts showed different trends between untreated cultures (control group) and osteogenic-induced cultures (treated group). In the control group, ERalpha mRNA climbed at peak levels at a confluence stage and decreased until day 20, whereas, in the treated group, the ERalpha mRNA kept climbing from a low level until day 20. Thus, the observed developmental expression of ERalpha mRNA correlates with progressive BMSCs growth and osteogenic differentiation and BMSCs may be a primary target cell for estrogen in maintaining bone formation.

[Study on the effect of gradually induced disordered occlusion on the condyle of New Zealand Rabbit TMJ].

OBJECTIVE: To investigate whether the gradually induced disordered occlusion can induce degenerative changes in rabbit TMJ. METHODS: 27 male New Zealand Rabbits, age of 5 months were divided equally and randomly into three groups: Group A: One premolar in upper dentition at one side and another premolar in lower dentition at the other side were pulled to move medially to the anterior intrinsic space by an orthodontic string to cause the corresponding premolars not to occlude with the opposites coincidently. Group B: the sham-operating controls, all animals were treated similar to those in Group A but without pulling. At the age of 6, 7 and 8 months, three rabbits of each group were killed by deeply anaesthetizing and bilateral condyles of TMJs were excised, sectioned and stained with HE. RESULTS: There was no significant histological difference in condylar cartilage between Group B and C. The cartilage of TMJs condyle in Group A was found with significant degenerative changes that increased with age. The thickness of cartilage of anterior and intermediate part of condyles decreased, the disturbance of continuation of hypertrophic layer appeared and part of hypertrophic layer was locally replaced by fibrous tissue. While the thickness of cartilage of posterior part of condyle increased obviously with a significant increase of the proportion of immature chondrocytes. CONCLUSION: The gradually induced disorder occlusion may lead to degenerative changes in rabbit condyle.