Preparation of ß-CD-Vitexin Microspheres and their Effects on SW480 Cell Proliferation.

OBJECTIVE: In order to overcome the insolution and low bioavailability of the vitexin in vivo, ß-cyclodextrin-vitexin (ß-CD-vitexin) microspheres were prepared, and their effects on the proliferation of SW480 cells were observed. METHODS: Scanning electron microscopy, ultraviolet spectrum, Fourier transform infrared spectroscopy, and release rate analysis identified the formation of ß-CD-vitexin microspheres. MTT assay detected the effect of ß-CD-vitexin microspheres on tumor cell proliferation at 6, 12, 24, and 48 h. Fluorescence microscopy and flow cytometry were used to observe the effect of ß-CD-vitexin microspheres on the apoptosis of SW480 cells. The mRNA expression of the p53 gene was measured by qPCR. RESULTS: ß-CD-vitexin microspheres were successfully prepared. SW480 cell proliferation was inhibited by 0.1, 0.2, and 0.4 mg/mL of ß-CD-vitexin microspheres in a dose- and time-dependent manner, and the mechanism of proliferation inhibition was related to cell apoptosis caused by the upregulated expression of p53 gene. CONCLUSION: The preparation of ß-CD-vitexin sustained release microspheres is feasible, and ß-CDvitexin microspheres have potential anti-colorectal cancer value.

Isorhamnetin inhibits progression of ovarian cancer by targeting ESR1.

Background: Although reports suggest Chinese herbal medicine treatment of ovarian cancer (OC) has a good effect, the role of isorhamnetin (ISO), a flavonol aglycone with immune, anti-inflammatory, cardiovascular and cerebrovascular protective effects, as well as an anticancer effect, in OC remains unclear. Network pharmacology was used to explore this in vitro and in vivo, and to identify relevant targets. Methods: The common targets of ISO in the treatment of OC were screened by constructing drug targets and disease gene databases for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. The protein-protein interaction network was constructed by STRING. Overlapping targets were further analyzed using the online tool UALCAN to analyze the correlation between gene expression and patient survival and prognosis. The effect of ISO on OC cell proliferation, migration, and invasion was assessed in vivo and in vitro, and the function of the estrogen receptor 1 (ESR1) in the development of OC was examined by overexpressing and knocking down ESR1 expression. Results: Through network pharmacology analysis, 25 target genes related to ISO-OC were screened out. The overall survival rate of OC patients only significantly correlated with high expression of ESR1 among 13 highly expressed overlapping genes. ISO significantly inhibited the proliferation, migration and invasion of OC cells in vitro and inhibited tumor growth in vivo. Overexpression of ESR1 significantly promoted the proliferation, migration and invasion of OC cells, whereas knockdown of ESR1 showed the opposite result. In addition, overexpression of ESR1 significantly reversed the inhibitory effect of ISO on the proliferation, migration and invasion of OC cells. Conclusions: We confirmed that ISO inhibits OC cell proliferation, migration and invasion by targeting ESR1 expression, which provides a theoretical basis for further pharmacological research.

Conserved RNA secondary structure in Cherry virus A 5'-UTR associated with translation regulation.

BACKGROUND: A variety of cis-acting RNA elements with structures in the 5'- or 3'-untranslated region (UTR) of viral genomes play key roles in viral translation. Cherry virus A (CVA) is a member of the genus Capillovirus in the family Betaflexiviridae. It has a positive single-stranded RNA genome of ~ 7400 nucleotides (nt). The length of the CVA 5'-UTR is ~ 100 nt; however, the function of this long UTR has not yet been reported. METHODS: Molecular and phylogenetic analyses were performed on 75 CVA sequences, which could be divided into four groups, and the RNA secondary structure was predicted in four CVA 5'-UTR types. These four CVA 5'-UTR types were then inserted upstream of the firefly luciferase reporter gene FLuc (FLuc), and in vitro translation of the corresponding transcripts was evaluated using wheat germ extract (WGE). Then, in-line structure probing was performed to reveal the conserved RNA structures in CVA-5'UTR. RESULTS: The four CVA 5'-UTR types appeared to have a conserved RNA structure, and the FLuc construct containing these four CVA 5'-UTR types increased the translation of FLuc by 2-3 folds, suggesting weak translation enhancement activity. Mutations in CVA 5'-UTR suppressed translation, suggesting that the conserved RNA structure was important for function. CONCLUSION: The conserved RNA secondary structure was identified by structural evolution analysis of different CVA isolates and was found to regulate translation.

Protective effect of hawthorn vitexin on the ethanol-injured DNA of BRL-3A hepatocytes.

ABSTRACT: Vitexin is a natural active ingredient in hawthorn leaves, which has a wide range of anti-tumor effects. This study was conducted to assess the protective effect of hawthorn vitexin on the ethanol-injured DNA of hepatocytes in vitro and to explore its mechanism. The effect of different concentrations of hawthorn vitexin on ethanol-injured hepatocytes was detected via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method to study the protective effect of hawthorn vitexin on ethanol-injured DNA damage in hepatocytes. Single-cell gel electrophoresis was used to observe the effect of hawthorn vitexin on ethanol-induced DNA damage in hepatocytes, and the Olive tail moment was measured. Cell physiological and biochemical indexes, such as superoxide dismutase activity, malonaldehyde content, and glutathione peroxidase activity, were detected with kits. The mRNA expression of the superoxide dismutase gene was measured via real-time quantitative polymerase chain reaction. It was showed that 0.2, 0.4, and 0.8 mg mL-1 hawthorn vitexin could significantly repair hepatocyte growth and ethanol-induced DNA damage. This effect was closely related to the improvement in superoxide dismutase, malonaldehyde, and glutathione peroxidase. Hawthorn vitexin could be used to repair ethanol-injured hepatocytes through antioxidation effects, and showed potential for the treatment of liver injury.

The association between overweight and obesity on bone mineral density in 12 to 15 years old adolescents in China.

ABSTRACT: Overweight/obesity can influence bone mineral accretion, but the conclusions are not consistent. We aimed to examine the association between bone mineral density (BMD) levels and body mass index (BMI) in 12 to 15 years old adolescents.We performed a cross-sectional study including 8365 adolescents. BMD was evaluated using a quantitative ultrasound device. Z scores for BMI were evaluated using World Health Organization references. Logistic regression models were performed to evaluate the association between BMD levels and BMI.Totally 1866 (22.3%) adolescents had low /reduced BMD, and boys had a higher rate than girls (72.6% vs 27.4%, P < .001). The rates of thinness, normal weight, overweight, and obesity were 2.8%, 57.1%, 22.3%, and 17.8%, respectively. The multivariable-adjusted (age, sex, systolic blood pressure, and height Z score) ORs (95% CIs) of low/reduced BMD associated with BMI groups (thinness, normal [reference], overweight, and obesity) were 0.59 (0.39-0.89), 1.00, 1.61 (1.41-1.84), and 1.98 (1.69-2.30), respectively (Ptrend < .001). This positive association existed in boys and girls though the differences were not significant between normal weight and thin girls. The multivariable-adjusted ORs for each 1-unit increase in BMI Z score were 1.36 (1.24-1.49) for girls, and 1.23 (1.16-1.30) for boys, and 1.26 (1.20-1.32) for all participants.We observed a positive association between BMI and low/reduced BMD in 12 to 15 years old adolescents. More attention should be paid on overweight and obese adolescents to reduce the risk of low BMD. Further studies are needed to explore the mechanisms of this association.

Overexpression of isochorismate synthase enhances salt tolerance in barley.

Isochorismate synthase (ICS) is a key enzyme for the synthesis of salicylic acid (SA) in plants. SA plays an important role in the response of plants to abiotic stress. In this study, transgenic barley was constructed to evaluate the function of ICS under salt stress. ICSOE lines showed obvious salt stress tolerance, this results from the increased outward Na+ flux and inward K+ flux in roots, thereby maintaining a lower cytosolic Na+/K+ ratio under salt stress. Overexprssion of ICS also improved Na+ sequestration in shoots under salt stress. In addition, ICSOE lines displayed less accumulation of reactive oxygen species and oxidative damage, accompanied by higher activity of antioxidant enzymes. The improved Na+/K+ ratio, Na+ sequestration, and antioxidative competence play an important role in the enhanced salt tolerance of ICSOE lines. These findings help to elucidate the abiotic stress resistance of the ICS pathway in barley.

Overexpression of isochorismate synthase enhances drought tolerance in barley.

Isochorismate synthase (ICS) is a key enzyme for the synthesis of salicylic acid (SA) in plants. SA mediates plant responses to both biotic and abiotic stresses. In previous studies, we found that overexpression of ICS (ICSOE) or suppression of ICS (ICSRNAi) affected the host response to Fusarium graminearum in barley. However, whether the barley ICS gene plays a role in adapting to abiotic stresses remains to be determined. In the present study, expression of the ICS gene was upregulated when treated with 20 % PEG6000, and ICSOE lines were more drought tolerant than wild type (WT) and ICSRNAi. In addition, the abscisic acid (ABA) levels in the ICSOE lines were higher than those in the WT and ICSRNAi lines under drought stress. High ABA levels significantly reduced Gs and E, which may impact water retention under drought stress. Under drought conditions, the activity of antioxidant enzymes was significantly higher in the ICSOE lines, correlating with a lower levels of reactive oxygen species (ROS) and malondialdehyde (MDA). Enhanced antioxidant competence also contributed to drought tolerance in ICSOE lines. These findings help elucidate the abiotic stress resistance of the ICS pathway in barley.

The impact of height on low/reduced bone mineral density in Chinese adolescents aged 12-14 years old: gender differences.

Low/reduced bone mineral density (BMD) is an important predictor of childhood fracture. In this article, we presented the prevalence of BMD in Chinese adolescents and, for the first time, demonstrated the gender disparities in the impact of height on BMD. PURPOSE: To analyze the gender disparities in the association of low/reduced BMD with height in Chinese adolescents at the stage of growth spurt. METHODS: A total of 8152 adolescents aged 12-14 years old were included based on a cross-sectional study in Tianjin, China. Height and weight were measured with standard equipment. BMD was measured using the method of quantitative ultrasound. Adolescents with Z ≤ - 2.0 or - 2.0 < Z ≤ - 1.0 were defined as "low BMD" or "reduced BMD". RESULTS: The total low/reduced BMD rate was 22.0% in Chinese adolescents aged 12-14 years old, and boys were more likely to have low/reduced BMD than girls (30.1% vs. 12.9%, P < 0.001). The rate of low/reduced BMD significantly increased with age in boys (Ptrend = 0.019), whereas decreased with age in girls (Ptrend = 0.018). We found significant interaction effect between gender and height standard deviation score (height-Z) in the association with low/reduced BMD (Pinteraction < 0.001). There was a positive association of height-Z among boys (OR = 1.30, 95%CI 1.21-1.39, P < 0.001), meanwhile low/reduced BMD was inversely associated with height-Z among girls (OR = 0.85, 95%CI 0.78-0.94, P < 0.001). CONCLUSIONS: Our study suggested strong gender disparities in the impact of height on BMD in Chinese adolescents aged 12-14 years old, where the association between low/reduced BMD and height was positive among boys but inverse among girls. The study provides evidence on the early prevention and the risk factor identification of low/reduced BMD and childhood fractures.

Curcumin Promotes Apoptosis of Activated Hepatic Stellate Cells by Inhibiting Protein Expression of the MyD88 Pathway.

Activation and proliferation of hepatic stellate cells (HSC) play an important role in the progress of liver fibrosis. HSC activation occurs in response to inflammatory cytokines, cellular interactions with immune cells, and morphogenetic signals. The literature hints to a role of the adaptor protein MyD88 in fibrosis. Although curcumin has been shown to exert inhibitory effects on the proliferation of HSC in vitro, its influence on the MyD88 pathway in HSC has remained unclear. Here, we investigated whether curcumin accelerates apoptosis of HSC through the MyD88 pathway. HSC (rat HSC T6) were divided into a control group, MyD88 small interfering RNA (siRNA) group, curcumin group, and curcumin + MyD88 siRNA group. The MyD88 siRNA groups were exposed to siRNA for 48 h. The curcumin groups were cultured in the presence of curcumin for 24 h. Apoptosis was detected by flow cytometry. For Toll-like receptor (TLR) 2 and 4 as well as MyD88 and the dependent factors NF-κB, TNF-α, and IL-1ß, mRNA expression was detected by reverse transcription polymerase chain reaction (RT-PCR). For MyD88, protein expression was further observed by Western Blot. Both curcumin and MyD88 siRNA inhibited the mRNA expression of MyD88 pathway-related effectors (TLR2, TLR4, NF-κB, TNF-α, IL-1ß) in HSC. Furthermore, both treatments reduced the expression of MyD88 protein in HSC and promoted their apoptosis. These effects were more obvious in the curcumin + MyD88 siRNA group. This study demonstrates that curcumin promotes apoptosis of activated HSC by inhibiting the expression of cytokines related to the MyD88 pathway. It elucidates the possible mechanisms of curcumin in inducing apoptosis of HSC through the MyD88 pathway.

Physical fitness status of children and adolescents in Tianjin of China during past three decades: a cross-sectional study.

BACKGROUND: Significant changes in the society and lifestyle have been observed in China, which influences the physical fitness status of children and adolescents. This study aimed to estimate the physical fitness status in Tianjin during the past three decades. METHODS: Cross sectional survey was used in this study. The data were derived from the Chinese National Students' Physical Health Survey database (1985-2010) using stratified cluster randomized sampling method. A total of 58,006 subjects aged 7-21 were recruited. The ethics approval was obtained from the Ethics Committee of Tianjin Medical University before collecting data from the children and adolescents, and/or their parents/guardians on behalf of them. Also written informed consent was obtained before collecting data from the patients. The secular trend of the physical fitness status, Levels of height, weight, body mass index, and chest circumference in China were calculated. The increase velocity of according indexes were analyzed, gender and rural/urban difference were discussed afterwards. SPSS16.0 was used in the analysis. RESULTS: Levels of height, weight, body mass index, and chest circumference were obviously increased in 2010 compared to those levels in 1985, especially in the 9-13 age group. It was observed that the increase in the two crosses for height velocity between two genders of rural students was disappeared and the age of seven was important for the physical development. CONCLUSIONS: The circumstance was good for the physical development of the children and adolescents. The disappearance of the two crosses for height velocity between two genders of rural students in this study might be associated with factors such as nutritional status.

Effect of acupuncture treatment on vascular cognitive impairment without dementia: study protocol for a randomized controlled trial.

BACKGROUND: Vascular cognitive impairment, no dementia (VCIND) is a condition at risk for future dementia and should be the target of preventive strategies. Preliminary evidence suggests that acupuncture may be a clinically effective intervention for people with early-stage vascular cognitive impairment. We will do a multicenter, 6-month, drug-controlled, nonblinded, randomized, parallel-group trial to determine whether acupuncture is effective for improving cognitive function and quality of life for patients with VCIND. METHODS/DESIGN: A total of 216 eligible patients will be recruited and randomly assigned acupuncture for two sessions/week (n = 108) or citicoline 300 mg/day (n = 108) in a multicenter, 6-month trial. The primary endpoint is cognition (Alzheimer's Disease Assessment Scale, Cognitive Subscale (ADAS-cog)). Secondary endpoints include assessments of activities of daily living and behavioral symptoms (Clock Drawing Test (CDT), Activities of Daily Living (ADL) and Instrumental Activities of Daily Living scale (IADL)). DISCUSSION: This will be the first large-scale trial specifically evaluating acupuncture therapy in VCIND. If the study confirms the effectiveness and safety of acupuncture treatment, it will be important to examine how the acupuncture approach could most effectively be integrated into the provision of routine healthcare. TRIAL REGISTRATION: This study is registered as an International Standard Randomised Controlled Trial on 17 January 2014, number ISRCTN 82980206.

Transforming growth factor­ß1 reduces apoptosis via autophagy activation in hepatic stellate cells.

Autophagy is a metabolic process that is important in fibrogenesis, in which cellular components are degraded by lysosomal machinery. Transforming growth factor ß1 (TGF­ß1) is a potent fibrogenic cytokine involved in liver fibrosis; however, it remains elusive whether autophagy is regulated by TGF­ß1 in this process. In the present study, the function of TGF­ß1­mediated autophagy in the proliferation and apoptosis of hepatic stellate cells (HSCs) was investigated. A rat HSC cell line (HSC­T6) was incubated with or without TGF­ß1 followed by bafilomycin A1, and microtubule-associated proteins 1A/1B light chain 3 (LC3) small interfering (si)RNA was used to inhibit autophagy in order to assess the association between TGF­ß1 and autophagy. HSC­T6 cell transient transfection was accomplished with a pLVX­AcGFP­N1­rLC3B­encoding plasmid. An MTS assay and flow cytometry were utilized to detect proliferation and apoptosis of HSC­T6 cells. Quantitative polymerase chain reaction, immunofluorescence and western blot analysis were used to detect the presence of activation markers. Proliferation was increased and apoptosis was reduced in HSC­T6 cells treated with TGF­ß1 compared with cells subjected to serum deprivation. However, when HSC­T6 cells were treated with bafilomycin A1 and LC3 siRNA, increased apoptosis and reduced proliferation were observed. In addition, protein and mRNA expression levels of the autophagy marker LC3 were significantly increased. GFP­LC3 punctate markings were more prolific following TGF­ß1 treatment of HSC­T6 cells, indicating that TGF­ß1 may rescue HSC­T6 cells from serum deprivation and reduce apoptosis via autophagy induction. The present study elucidated the possible functions of TGF­ß1­mediated autophagy in the pathological process of liver fibrosis.

ß-Nerve growth factor attenuates hepatocyte injury induced by D-galactosamine in vitro via TrkA NGFR.

Nerve growth factor (NGF) regulates the proliferation, differentiation and survival of cells and is also involved in the wound healing and tissue remodeling processes. The biological effects of NGF are dependent upon receptor signal-mediating functions, which differ between cells. This study attempted to investigate the hepatoprotective effect and possible mechanism of ß-NGF on D-galactosamine (D-GalN)-injured human liver L-02 cell lines. We demonstrated that L-02 cells expressed the neurotrophin receptors tyrosine kinase-A nerve growth factor receptor (TrkA NGFR) and p75 pan-neurotrophin receptor (p75NTR). Recombinant human ß-NGF markedly reduced cell injury and promoted the proliferation of L-02 cells damaged by D-GalN. However, this proliferation effect was blocked by the anti-TrkA NGFR antibody. Lactate dehydrogenase (LDH) and malondialdehyde (MDA) were released at reduced levels in the L-02 cell culture supernatant pretreated with ß-NGF. Furthermore, the albumin (ALB) content in the cell medium and intracellular glutathione (GSH) levels were markedly augmented, and the permeability of the mitochondrial membrane of the L-02 cells was improved by ß-NGF. Our results suggested that exogenous ß-NGF protects L-02 cells from D-GalN-induced injury through the NGF/TrkA NGFR signaling pathway.

[Expression of eotaxin mRNA and TNF-alpha mRNA in lung tissues of sensitized mice and modulation by anti-inflammatory drugs].

OBJECTIVE: To establish determination methods of eotaxin mRNA and TNF-alpha mRNA expression in the lung tissue of mice. METHODS: Eotaxin mRNA and TNF-alpha mRNA expressions were determined by semi-quantitative RT-PCR. The functional implications of eotaxin mRNA and TNF-alpha mRNA expression were examined by detecting the numbers of total leucocytes and eosinophils in bronchoalveolar lavage fluid(BALF). RESULT: Eotaxin mRNA and TNF-alpha mRNA expression in lung tissue total numbers of leucocyte and numbers of eosinophil in BALF increased in sensitized mice compared with those in normal mice. Dexamethasone significantly but did not inhibit eotaxin mRNA and TNF-alpha mRNA expressions, and eosinophil infiltration in the lungs of the sensitized mice. A compound preparation of traditional Chinese medicine inhibited eotaxin mRNA and eosinophil infiltration, influenced TNF-alpha mRNA expression. CONCLUSION: Increased eotaxin mRNA expression in lung tissue is associated with eosinophil infiltration in BALF, which indicates that the methods of semi-quantitative RT-PCR may be useful to the study of the mechanism of antiasthmatic medicine.