ABSTRACT
To compare the pancreatic proteomics and autophagy between Rehmanniae Radix-and Rehmanniae Radix Praeparata-treated mice with type 2 diabetes mellitus(T2DM). The T2DM mouse model was established by high-fat diet coupled with streptozotocin(STZ, intraperitoneal injection, 100 mg·kg~(-1), once a day for three consecutive days). The mice were then randomly assigned into a control group, low-(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix groups, low-(150 mg·kg~(-1)) and high-dose(300 mg·kg~(-1)) catalpol groups, low-(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix Praeparata groups, low-(150 mg·kg~(-1)) and high-dose(300 mg·kg~(-1)) 5-hydroxymethyl furfuraldehyde(5-HMF) groups, and a metformin(250 mg·kg~(-1)) group. In addition, a normal group was also set and each group included 8 mice. The pancreas was collected after four weeks of administration and proteomics tools were employed to study the effects of Rehmanniae Radix and Rehmanniae Radix Praeparata on protein expression in the pancreas of T2DM mice. The expression levels of proteins involved in autophagy, inflammation, and oxidative stress response in the pancreatic tissues of T2DM mice were determined by western blotting, immunohistochemical assay, and transmission electron microscopy. The results showed that the differential proteins between the model group and Rehmanniae Radix/Rehmanniae Radix Prae-parata group were enriched in 7 KEGG pathways, such as autophagy-animal, which indicated that the 7 pathways may be associated with T2DM. Compared with the control group, drug administration significantly up-regulated the expression levels of beclin1 and phosphorylated mammalian target of rapamycin(p-mTOR)/mTOR and down-regulated those of the inflammation indicators, Toll-like receptor-4(TLR4) and Nod-like receptor protein 3(NLRP3), in the pancreas of T2DM mice, and Rehmanniae Radix showed better performance. In addition, the expression levels of inducible nitric oxide synthase(iNOS), nuclear factor erythroid 2-related factor 2(Nrf2), and heine oxygenase-1(HO-1) in the pancreas of T2DM mice were down-regulated after drug administration, and Rehmanniae Radix Praeparata demonstrated better performance. The results indicate that both Rehmanniae Radix and Rehmanniae Radix Praeparata can alleviate the inflammatory symptoms, reduce oxidative stress response, and increase the autophagy level in the pancreas of T2DM mice, while they exert the effect on different autophagy pathways.
Subject(s)
Diabetes Mellitus, Type 2 , Mice , Animals , Diabetes Mellitus, Type 2/drug therapy , Diabetes Mellitus, Type 2/genetics , Streptozocin/pharmacology , Diet, High-Fat/adverse effects , Proteomics , Inflammation , TOR Serine-Threonine Kinases , Autophagy , MammalsABSTRACT
BACKGROUND: Cardiovascular disease is one of the major threats to human life and health, and vascular aging is an important cause of its occurrence. Antisense non-coding RNA in the INK4 locus (ANRIL) is a kind of long non-coding RNA (lncRNA) that plays important roles in cell senescence. However, the role and mechanism of ANRIL in senescence of vascular smooth muscle cells (VSMCs) are unclear. METHODS: Cell viability and cell cycle were evaluated using an MTT assay and flow cytometry analysis, respectively. Senescence-associated (SA)-ß-galactosidase (gal) staining was used to determine cell senescence. Dual luciferase reporter assays were conducted to confirm the binding of ANRIL and miR-181a, as well as miR-181a and Sirt1. The expression of ANRIL, miR-181a, and Sirt1 was determined using qRT-PCR and protein levels of SA-ß-gal and p53-p21 pathway-related proteins were evaluated by Western blotting. RESULTS: ANRIL and Sirt1 were down-regulated, whereas miR-181a was up-regulated in aging VSMCs. In young and aging VSMCs, over-expression of ANRIL could down-regulate miR-181a and up-regulate Sirt1. MTT and SA-ß-gal staining assays showed that over-expression of ANRIL and inhibition of miR-181a promoted cell viability and inhibited VSMC senescence. The dual-luciferase reporter assay determined that miR-181a directly targets ANRIL and the 3'-UTR of Sirt1. Furthermore, over-expression of ANRIL inhibited cell cycle arrest and the p53-p21 pathway. CONCLUSION: ANRIL promotes cell viability and inhibits senescence in VSMCs, possibly by regulating miR-181a/Sirt1, and alleviating cell cycle arrest by inhibiting the p53-p21 pathway. This study provides novel insights for the role of ANRIL in the development of cell senescence.
Subject(s)
Cellular Senescence/drug effects , MicroRNAs/antagonists & inhibitors , Muscle, Smooth, Vascular/drug effects , RNA, Long Noncoding/pharmacology , Sirtuin 1/antagonists & inhibitors , Cell Cycle/drug effects , Cell Survival/drug effects , Cells, Cultured , Down-Regulation/drug effects , Humans , MicroRNAs/metabolism , Muscle, Smooth, Vascular/metabolism , Signal Transduction/drug effects , Sirtuin 1/metabolismABSTRACT
BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) exhibits poor prognosis and resistance to chemotherapy. This study was to identify the biomarkers associated with the progression, poor prognosis and chemoresistance of PDAC. METHODS: miR-34a and miR-150 levels in the plasma and tissues from PDAC patients were measured by real-time PCR. Xenograft PDAC tumor models were established in mice by inoculation of CD133+ stem cells isolated from PDAC tumors. Protein expression was measured by Western blot. RESULTS: The plasma miR-34a and miR-150 levels were significantly lower in PDAC patients than in patients with benign pancreatic lesions and in healthy subjects. The miR-34a and miR-150 levels in the tumor tissues were significantly lower than in pancreatic tissues with benign lesions. The protein levels of CD133, Notch1, Notch2 and Notch4 receptors in PDAC tumor tissues were significantly higher than in pancreatic tissues with benign lesions. miR-34a injection significantly inhibited the tumor growth of PDAC tumors and sensitized the anticancer effects of 5-fluorouracil (5-FU). miR-34a significantly inhibited Notch1, Notch2 and Notch4 expression in xenograft tumor tissues in vivo and BxPC-3 cells in vitro. miR-34a and miR-150 significantly induced apoptosis and inhibited proliferation, invasion and migration in BxPC-3 cells. miR-34a, but not miR-150, significantly sensitized the anticancer effect of 5-FU in BxPC-3 cells in vitro. CONCLUSION: A loss of expression of miR-34a, but not of miR-150, is associated with disease progression and poor prognosis in PDAC patients, and may be involved in the chemoresistance of PDAC cells.
Subject(s)
Biomarkers, Tumor/metabolism , Carcinoma, Pancreatic Ductal/genetics , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , Pancreatic Neoplasms/genetics , Adult , Aged , Animals , Carcinoma, Pancreatic Ductal/diagnosis , Cell Line, Tumor , Disease Progression , Female , Humans , Male , Mice , Mice, Inbred BALB C , Middle Aged , Pancreas/metabolism , Pancreas/pathology , Pancreatic Neoplasms/diagnosis , Prognosis , Random Allocation , Pancreatic NeoplasmsABSTRACT
Rapid restoration of perfusion in ischemic myocardium is the most direct and effective treatment for coronary heart disease but may cause myocardial ischemia/reperfusion injury (MIRI). Cinnamaldehyde (CA, C9H8O), a key component in the well-known Chinese medicine cinnamomum cassia, has cardioprotective effects against MIRI. This study aimed to observe the therapeutic effect of CA on MIRI and to elucidate its potential mechanism. H9C2 rat cardiomyocytes were pretreated with CA solution at 0, 10, and 100 µM, respectively and subjected to oxygen-glucose deprivation/reoxygenation (OGD/R). Then the cell viability, the NF-κB and caspase3 gene levels, the reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio, superoxide dismutase (SOD) level, reactive oxygen species (ROS) generation, 4-hydroxynonenal (4-HNE), and malondialdehyde (MDA) were detected. The severity of DNA damage was assessed by tail moment (TM) values using alkaline comet assay. Besides, the DNA damage-related proteins and the key proteins of the Nrf2 pathway were detected by western blot. CA treatment increased the cell viability, GHS/GSSG ratio, SOD level, PARP1, Nrf2, PPAR-γ, and HO-1 protein levels of H9C2 cardiomyocytes, while reducing NF-κB, caspase3, ROS level, 4-HNE and MDA content, γ-H2AX protein level, and TM values. Inhibition of the Nrf2 pathway reversed the effect of CA on cell viability and apoptosis of OGD/R induced H9C2 cardiomyocytes. Besides, 100 µM CA was more effective than 10 µM CA. In the OGD/R-induced H9C2 cardiomyocyte model, CA can protect cardiomyocytes from MIRI by attenuating lipid peroxidation and repairing DNA damage. The mechanism may be related to the activation of the Nrf2 pathway.
Subject(s)
Acrolein , Myocytes, Cardiac , NF-E2-Related Factor 2 , Oxygen , Animals , Rats , Acrolein/analogs & derivatives , Acrolein/pharmacology , Apoptosis , DNA Damage , Glucose/pharmacology , Glutathione Disulfide/genetics , Glutathione Disulfide/metabolism , Glutathione Disulfide/pharmacology , Lipid Peroxidation , Myocytes, Cardiac/drug effects , Myocytes, Cardiac/metabolism , NF-E2-Related Factor 2/drug effects , NF-E2-Related Factor 2/metabolism , NF-kappa B/metabolism , Oxidative Stress , Oxygen/metabolism , Reactive Oxygen Species/metabolism , Superoxide Dismutase/metabolismABSTRACT
This study investigated the expression of interleukin-17 (IL-17) and T cell immunoglobulin mucin and domain-containing molecule-3 (Tim-3) in bronchoalveolar lavage fluid (BALF) of asthmatic mice and the effect of dexamethasone (DEX) on these factors. Thirty-six mice were randomly divided into three groups: normal group, asthmatic group and DEX group. The mouse model of asthma was established by sensitization with ovalbumin in both the asthmatic and DEX groups. The levels of IL-6, IL-10, IL-17 and TGF-ß were measured in BALF by enzyme-linked immunesorbent assay (ELISA). The mRNA expression level of Tim-3 was detected by reverse transcription polymerase chain reaction (RT-PCR). The ratio of Tim-3+CD4+ cells to total CD4+ cells in BALF was determined by flow cytometry. Differential inflammatory cells in BALF were detected. The correlations among IL-17, IL-6, IL-10, Tim-3 and inflammatory cells were analyzed. The results showed that the levels of IL-17, IL-6 and Tim-3 were substantially increased and the IL-10 level decreased in BALF in the asthmatic mice, which was significantly reversed by DEX treatment. IL-17 expression was positively correlated with IL-6 and Tim-3 expression and the number of inflammatory cells but negatively with IL-10 expression. These results indicate that the increased expression of IL-17 and Tim-3 in BALF may be implicated in the occurrence and development of asthmatic inflammation; the mechanism by which DEX suppresses asthmatic airway inflammation involves down-regulation of IL-17 and Tim-3 levels.
Subject(s)
Asthma/metabolism , Dexamethasone/pharmacology , Gene Expression/drug effects , Interleukin-17/metabolism , Receptors, Virus/metabolism , Animals , Asthma/drug therapy , Asthma/genetics , Bronchoalveolar Lavage Fluid/chemistry , Female , Gene Expression/genetics , Hepatitis A Virus Cellular Receptor 2 , Interleukin-17/genetics , Mice , Mice, Inbred BALB C , Receptors, Virus/geneticsABSTRACT
OBJECTIVE: To reveal protein diversity between the same batch of fresh Houttuynia cordata in the same GAP base,and to lay the foundation construction for "node metabolic network". METHODS: Three methods including the Ramagli improved Bradford law, SDS-PAGE gel electrophoresis method and double wavelength thin-layer scanning method were used to study the total protein content diversity, protein species diversity and various kinds of content variability. RESULTS: The molecular weight of 53 plant protein mostly concentrated in the range of 6.5-97.2 kDa, the species diversity was not obvious with main performance for banding color shades; The RSD of zero moment (AUCT), first moment (MCRTT) and second moment (VCRTT) in protein electrophoresis banding was 40.92%, 6.01% and 18.57%, respectively. CONCLUSION: There is rich diversity in different Houttuynia cordata plant in the same GAP base, which provides basis for the foundation of subsequent key protease search, "node metabolic network" construction, and study on the Chinese medicine quality stability.
Subject(s)
Drugs, Chinese Herbal/chemistry , Houttuynia/chemistry , Plant Proteins/chemistry , Electrophoresis, Polyacrylamide Gel , Genetic Variation , Houttuynia/genetics , Quality ControlABSTRACT
OBJECTIVE: The carotid body (CB) is a major peripheral respiratory chemoreceptor. In patients with obstructive sleep apnea (OSA), high CB chemosensitivity (CBC) is associated with refractory hypertension and insulin resistance and known to further aggravate OSA. Thus, the identification of high CB (hCBC) among OSA patients is of clinical significance, but detection methods are still limited. Therefore, this study aimed to explore the association of CBC with OSA severity and to develop a simplified model that can identify patients with hCBC. METHODS: In this cross-sectional study of subjects who underwent polysomnography (PSG), CBC was measured using the Dejours test. We defined hCBC as a decrease of >12% in respiratory rate (RR) after breathing of pure O2. The association of CBC with OSA severity was explored by logistic regression, and a model for identifying hCBC was constructed and confirmed using receiver operating characteristic analysis. RESULTS: Patients with OSA (n=142) and individuals without OSA (n=38) were enrolled. CBC was higher in patients with OSA than in those without OSA (% decrease in RR, 15.2%±13.3% vs 9.1%±7.5%, P<0.05). Apnea-hypopnea index (AHI), fraction of apnea-hypopnea events in rapid-eye-movement sleep (Fevents-in-REM), and longest time of apnea (LTA) were associated with hCBC independently (odds ratio [OR]=1.048, OR=1.082, and OR=1.024 respectively; all P<0.05). The model for identifying hCBC allocated a score to each criterion according to its OR values, ie, 1 (LTA >48.4 s), 2 (AHI >15.7 events/hour), and 3 (Fevents-in-REM >12.7%). A score of 3 or greater indicated hCBC with a sensitivity of 79.4% and specificity of 88.2%. CONCLUSION: High CBC is associated with the severity of OSA. A simplified scoring system based on clinical variables from PSG can be used to identify hCBC.
ABSTRACT
Despite progress achieved in cancer chemotherapy in recent decades, adverse effects remain a limiting factor for a number of patients with colorectal cancer, suggesting the requirement for novel therapeutic strategies. Gene therapy appears to be a promising strategy for treating cancer. The present study aimed to investigate the anti-tumor effect of a combined gene therapy, using Survivin downregulation by RNAi and a fusion suicide gene yCDglyTK therapy system. A triple-gene vector expressing Survivin-targeted small hairpin RNA (Survivin-shRNA) and fusion suicide gene yCDglyTK was constructed, and administered to HCT116 cells. Survivin expression decreased significantly and yCDglyTK fusion gene expression was confirmed by both reverse transcription-quantitative polymerase chain reaction and western blot analysis. Introduction of Survivin-shRNA into yCDglyTK/prodrug system eradicated colon cancer cells and induced apoptosis more effectively. Furthermore, this therapeutic system is able to inhibit the migration of HCT116 cells. These results indicate that the recombinant plasmid may serve as a novel gene therapy approach to treat colorectal carcinoma.
ABSTRACT
Lipopolysaccharides (LPSs) are an important initiation factor in acute respiratory distress syndrome. The aim of the present study was to investigate the effect of LPSs on the regulation of angiotensin II (Ang II) receptors in human pulmonary microvascular endothelial cells (HPMECs). HPMECs were treated with 0, 50, 100 or 200 ng/ml LPS and incubated for 4, 8, 12 or 16 h. The expression of Ang II type 1 receptor (AT1R) and Ang II type 2 receptor (AT2R) was determined using reverse transcriptionpolymerase chain reaction and western blot analysis. The affinity to Ang II was measured using a radioligand binding assay. No AT2R expression was detected with or without LPS administration in HPMECs, and LPS treatment increased the expression level of AT1R. A timedependent increase of AT1R transcription was observed in the 50 ng/ml group, while in the 100 and 200 ng/ml groups, the AT1R mRNA levels reached peak values at 4 h and remained unchanged. The protein level of AT1R increased significantly in a dosedependent manner for each incubation time period. A timedependent increase in the protein level was observed in the 50 and 100 ng/ml groups. As for the 200 ng/ml group, the level of AT1R reached a peak value at 8 h. Maximal binding (Bmax) significantly increased following LPS treatment and Bmax of the 50 ng/ml group exhibited a timedependent increase. The Bmax of the 100 and 200 ng/ml groups reached peak values at 12 and 8 h, respectively, and decreased thereafter. The dissociation constant remained unchanged following LPS treatment. Thus, treatment with LPS increased AT1R expression and its Bmax to Ang II in HPMECs, however, did not alter the affinity of AT1R to Ang II.
Subject(s)
Endothelial Cells/drug effects , Lipopolysaccharides/pharmacology , Receptor, Angiotensin, Type 1/metabolism , Cells, Cultured , Endothelial Cells/metabolism , Humans , Receptor, Angiotensin, Type 1/physiology , Renin-Angiotensin SystemABSTRACT
AIM: To explore the possible mechanism why drinking Maotai liquor dose not cause hepatic fibrosis. METHODS: After being fed with Maotai for 56 days consecutively, the male SD rats were decollated for detecting the biological indexes, and the livers were harvested to examine the liver indexes and the level of hepatic metallothioneins (MT). Hepatic stellate cells (HSC) proliferation and collagen generation were also observed. RESULTS: Hepatic MT contents were 216.0 ng.g(-1)+/-10.8 ng.g(-1) in the rats of Maotai group and 10.0 ng.g(-1)+/-2.8 ng.g(-1) in the normal control group, which was increased obviously in Maotain group (P<0.05). In the rats with grade CCL(2) poisoning induced by Maotai, hepatic MT content was 304.8 ng.g(-1)+/-12.1 ng.g(-1) whereas in the controls with grade CCL(4) poisoning, it was 126.4 ng.g(-1)+/-4.8 ng.g(-1) (P<0.05). MDA was 102.0 nmol.g(-1)+/-3.4 nmol.g(-1) in Maotai group and 150.8 nmol.g(-1)+/-6.7 nmol.g(-1) in the control group (P<0.05). When both of the groups were suffering from grade CCL(4) poisoning, hepatic MT contents was negatively correlated with MDA (r=-0.8023, n=20, P<0.01). The 570 nmA values of each tube with HSC regeneration at concentrations of 0, 10, 50, 100, and 200 g.L(-1) of Maotai were 0.818, 0.742, 0.736, 0.72, 0.682, and 0.604, respectively. From the concentration of 10 g.L(-1), Maotai began to show obvious inhibitory effects against HSC, and the inhibition was concentration-dependent (P<0.05, P<0.01). Type I collagen contents in HSC were 61.4, 59.9, 50.1, 49.2, 48.7, 34.4 microg.g(-1) at concentrations of 0, 10, 50, 100, and 200 g.L(-1) of Maotai. At the concentration of 100-200 g.L(-1), Maotai had obvious inhibitory effect against the secretion of type I collagen (P<0.05). Gene expression analysis was conducted on cells with Maotai concentrations of 0, 50, 100g.L(-1) respectively and the ash values of beta-actin gene expression were 0.88, 0.74, and 0.59, respectively,suggesting that at the concentration of 100g.L(-1), Maotai could obviously inhibit gene expression of type I procollagen (P<0.05), but the effect was not obvious at the concentration of 50 g.L(-1) (P>0.05). At the concentration of 10 g.L(-1), HSC growth in vitro inhibition rates were 16.4+/-2.3 in Maotai group and -8.4+/-2.3 in the control group (P<0.05). CONCLUSION: Maotai liquor can increase metallothioneins in the liver and inhibit the activation of HSC and the synthesis of collagen in many aspects, which might be the mechanism that Maotai liquor interferes in the hepatic fibrosis.
Subject(s)
Alcoholic Beverages/toxicity , Hepatocytes/drug effects , Metallothionein/biosynthesis , Animals , Cell Division/drug effects , Collagen/biosynthesis , Hepatocytes/metabolism , Hepatocytes/pathology , Humans , Liver Cirrhosis, Alcoholic/etiology , Liver Cirrhosis, Alcoholic/prevention & control , Male , Rats , Rats, Sprague-DawleyABSTRACT
BACKGROUND: Epidemiology investigation showed that no worker drunk Maotai liquor for nearly 30 years died of hepatic diseases, and no obvious hepatic fibrosis and cirrhosis were found in 99 workers who had drunk Maotai liquor for a long period by epidemiology investigation and needle biopsy. The same finding was detected in rats that were drunk by Maotai liquor continued for 56 days. This study was to investigate the effects of Maotai liquor on the liver and its mechanism of preventing hepatic fibrosis. METHODS: After ingestion of Maotai for 56 consecutive days, male SD rats were killed for detecting the levels of metallothionein and malondialdehyde (MDA) in liver tissues. Rat hepatic stellate cells (HSCs) and human HSCs were cultured in vitro to observe the effect of Maotai on HSCs proliferation and collagen synthesis. After ingestion of Maotai for 14 consecutive weeks, the livers of male SD rats were harvested for pathohistological examination. RESULTS: The level of metallothionein in the liver of Maotai-induced rats increased by 22 folds, whereas the levels of hepatic lipid peroxide and MDA was decreased significantly (P<0.05) in Maotai-induced animals suffering from CCl4. Maotai demonstrated obvious inhibitory effect on proliferation of HSCs and the inhibition was concentration-dependent. Gene expression and protein secretion of collagens could also be inhibited by Maotai. In alcoholic group, typical liver cirrhosis was observed. In Maotai group, however, though fatty degeneration of hepatocytes and mild fibrosis of the interstitium were observed, no obvious hepatic fibrosis and cirrhosis were found. CONCLUSION: It might be an important mechanism of interfering the progress of hepatic fibrosis that Maotai increases the level of metallothionein in the liver and inhibits the activation of HSCs and the synthesis of collagen proteins.
Subject(s)
Alcoholic Beverages/toxicity , Hepatocytes/drug effects , Metallothionein/biosynthesis , Animals , Base Sequence , Biopsy, Needle , Cell Division/drug effects , Cells, Cultured , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Immunohistochemistry , Liver Cirrhosis, Alcoholic/etiology , Liver Cirrhosis, Alcoholic/prevention & control , Male , Molecular Sequence Data , Polymerase Chain Reaction , Probability , Rats , Rats, Sprague-Dawley , Reference Values , Risk Assessment , Sensitivity and SpecificityABSTRACT
OBJECTIVE: To explore the effect on liver and the mechanism of preventing hepatic fibrosis by drinking Kweichow Moutai liquor (Maotai). METHODS: (1) After ingested with Maotai for 56 days consecutively, the male SD rats were decollated for detecting metallothioneins and MDA content in liver tissues; (2) Culturing rat hepatic stellate cell (HSC) and human HSC in vitro, observing the effect of Maotai on HSC's proliferation and collagen synthesis; (3) After male SD rats were ingested with Maotai for 14 weeks consecutively, the livers were harvested for pathohistological examination. RESULTS: (1) Metallothioneins content in the liver of Maotai-induced rats increased by 22 folds, the production of hepatic lipid peroxide, MDA was significantly decreased (P < 0.05) in Maotai-induced animals suffering from CCL4; (2) Maotai demonstrate obvious inhibitory effect against proliferation of HSC, and the inhibition was concentration-dependent. gene expression and protein secretion of collagens could also be inhibited by Maotai; (3) In control alcoholic group, typical cirrhosis of liver was shaped. In Maotai group, however, though fatty degeneration of hepatocytes and mild fibrosis of interstitium were observed, no obvious hepatic fibrosis and cirrhosis were found. CONCLUSION: It might be an important mechanism of interfering hepatic fibrosis progressing that Maotai induces the increase of metallothioneins content in the liver, inhibits the activation HSC and the synthesis of collagen protein.
Subject(s)
Alcoholic Beverages/toxicity , Hepatocytes/drug effects , Metallothionein/biosynthesis , Animals , Cells, Cultured , Collagen/biosynthesis , Hepatocytes/metabolism , Hepatocytes/pathology , Humans , Liver Cirrhosis, Alcoholic/etiology , Liver Cirrhosis, Alcoholic/metabolism , Liver Cirrhosis, Alcoholic/prevention & control , Male , Metallothionein/analysis , Rats , Rats, Sprague-DawleyABSTRACT
OBJECTIVE: To investigate the effects of abnormal Savda Munziq (ASMq) total phenolics combined with cisplatin and docetaxel on the Hela cell growth. METHODS: In vivo cultured Hela cells were treated with cisplatin, docetaxel, total phenolics, cisplatin+total phenolics or docetaxel+total phenolics. MTT was performed to assess inhibition of cell proliferation, flow cytometry to detect apoptosis, and semi-quantitative RT-PCR to test for survivin and Bcl-2 expression. RESULTS: The total phenolics, cisplatin and docetaxel had significant inhibitory and apoptosis-promoting effects on Hela cells (P<0.05), with the early apoptotic rates of 12.8±0.70%, 18.9±3.79% and 15.8±3.8)%; the total phenolics, cisplatin and docetaxel significantly decreased the expression of Bcl-2 and survivin (all P<0.01), especially when used in combination. CONCLUSION: ASMq total phenolics, combined with cisplatin and docetaxel, could promote the apoptosis of Hela cells possibly through reducing the expression of Bcl-2 and survivin.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/pharmacology , Apoptosis/drug effects , Cell Proliferation/drug effects , Phenols/pharmacology , Plants, Medicinal/chemistry , Cisplatin/administration & dosage , Docetaxel , Drugs, Chinese Herbal/chemistry , Flow Cytometry , HeLa Cells , Humans , Inhibitor of Apoptosis Proteins/genetics , Inhibitor of Apoptosis Proteins/metabolism , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Survivin , Taxoids/administration & dosageABSTRACT
OBJECTIVE: The stable quality of Chinese herbal medicines is a critical factor for their reliable clinical efficiency. An improved liquid-liquid extraction procedure and a liquid chromatographic method were developed to simultaneously analyze five anthraquinones (aloe-emodin, rhein, emodin, chrysophanol and physcion) in a Chinese traditional hospital preparation, Fuyankang mixture, in order to quantitatively control its quality in a more effective way. METHODS: A more economical and repeatable extraction procedure based on conventional liquid-liquid extraction technique was developed and used to extract five marker components in Fuyankang mixture. These anthraquinones were separated in less than 20 min on a C18 column with methanol and 0.1% phosphoric acid (88:12, v/v) as mobile phase. The method was validated for specificity, precision, spiked recovery and stability. RESULTS: Compared to conventional liquid-liquid extraction, the improved liquid-liquid extraction was found to be more effective for simultaneous extraction of anthraquinones from an aqueous Chinese herbal preparation, especially for hydrophobic compounds. The improved extraction method was successfully applied to determine the content of five marker components in Fuyankang mixture by the means of reverse phase high-performance liquid chromatography. CONCLUSION: The improved extraction procedure may be suitable for routine quality control of Fuyankang mixture and other traditional preparations at city-level hospitals in China.
Subject(s)
Anthraquinones/analysis , Chromatography, High Pressure Liquid/methods , Chromatography, Reverse-Phase/methods , Drugs, Chinese Herbal/analysis , Emodin/analogs & derivatives , Emodin/analysisABSTRACT
BACKGROUND: Stroke occurs upon obstruction of cerebral blood circulation and is clinically characterized by sudden onset symptoms. Advanced age is the main risk factor of stroke, but cases of pediatric stroke have been rarely reported. This study aimed to determine the etiology, clinical presentation, and radiologic features of neurological deficit for pediatric arterial ischemic stroke (PAIS). METHODS: The medical records of 42 PAIS patients (age range: 9 months to 13 years) treated at Wuhan Children's Hospital between July 2007 and January 2011 were retrospectively reviewed. Infarction location was first determined by craniocerebral computed tomography and magnetic resonance (MR) imaging. The stenotic or occluded main cerebral arteries and/or branches were determined by MR angiography and digital subtraction angiography. RESULTS: The majority of the 42 PAIS cases (66.7%, n = 28) were ≤ 3 years old (vs. >3 years old: 33.3%, n = 14; P<0.05), but the male: female ratio was similar in both groups (P > 0.05). The most frequently reported signs and symptoms for both age groups were limited physical activity followed by convulsions and delirium, but convulsions were more prevalent in children ≤ 3 years-old. Children > 3 years-old mainly experienced the limited physical activity symptoms, including hemiparalysis, aphasia, and ataxia. For all 42 cases, the most frequent etiologies were infections (38.1%, n = 16), iron deficiency anemia (16.7%, n = 7), and moyamoya syndrome (11.9%, n = 5). The predominant infarcts among all cases were middle cerebral artery (63.6%, n = 21) and basal ganglia (64.3%, n = 27). CONCLUSIONS: PAIS occurs more frequently in younger children and this group most frequently presents with convulsion as the initial symptom. The overall etiologies of PAIS may be different from those of adult stroke and the involved regions may be distinguishing features of PAIS or its different forms, but more research is required.
Subject(s)
Stroke/diagnostic imaging , Stroke/etiology , Adolescent , Child , Child, Preschool , Female , Humans , Infant , Male , Radiography , Retrospective Studies , Stroke/diagnosisABSTRACT
Alkaline treatment is widely used for improving biogas production for lignocellulosic materials. This study was conducted to investigate the effect of alkaline treatment on physicochemical property of digested Spartina alterniflora (DSA). Fourier Transform Infrared Spectroscopy (FTIR), X-Ray diffraction patterns, proton Nuclear Magnetic Resonance Spectroscopy (1H-NMR), solid-state 13C-NMR Spectroscopy and some general indicators were used to analyze changes of the lignocellulosic structure and composition of NaOH-treated digested Spartina alterniflora. The results showed that, after NaOH treatment, surface lignin and some carbohydrate were destructed into lignin fragment, organic acids and some other small molecular organic matter, but the skeleton structure of lignin and cellulose of DSA were not destructed significantly. The crystalline of cellulose of DSA was transferred into biodegradable forms and content of crystalline of DSA and cellulose were increased after NaOH treatment. The results of 13C-NMR showed that methyl (CH3) and carboxylic C(COOH) groups of DSA were decomposed significantly.
Subject(s)
Chemical Phenomena/drug effects , Fermentation , Poaceae/metabolism , Sodium Hydroxide/chemistry , Anaerobiosis , Biodegradation, Environmental , Biofuels/analysis , Cellulose/metabolism , Poaceae/chemistryABSTRACT
OBJECTIVE: To establish a method for inducing apoptosis of rhesus peripheral blood lymphocytes (PBLs). METHODS: Rhesus PBLs were irradiated with X-ray, (60)Co gamma-rays and ultraviolet (UVC254 nm), respectively, and the cell apoptosis was evaluated with flow cytometry using annexin-V staining and propidium iodide staining. RESULTS: X-ray and (60)Co gamma-ray irradiation induced only low apoptotic rates of the PBLs, and UVC resulted in the highest apoptotic rate of about 60%. UVC irradiation of the PBLs in RPMI supplemented with 10% heat-inactivated fetal calf serum for 60 min at a distance of 20 cm led to an early apoptotic rate of 58.85% and necrotic rate of 11.5%. The apoptotic rate of PBLs increased in a dose- and time-dependent fashion. CONCLUSION: For inducing apoptosis of the rhesus PBLs, UVC can be more effective than X-ray and (60)Co gamma-ray. The highest apoptotic rate can be achieved when the rhesus PBLs in RPMI supplemented with 10% heat-inactivated fetal calf serum are exposed to UVC for 60 min at the distance of 20 cm.