Baicalin alleviates silica-induced lung inflammation and fibrosis by inhibiting TLR4/NF-?B pathway in rats.

Silicosis is an occupational lung disease caused by inhaling silica dust. The disease is characterized by early lung inflammation and late irreversible pulmonary fibrosis. Here we report the effect of Baicalin, a main flavonoid compound from the roots of Chinese herbal medicine Huang Qin on silicosis in a rat model. Results showed Baicalin (50 or 100 mg/kg/day) can mitigate the silica-induced lung inflammation and reduce the harm of alveolar structure and the blue region of collagen fibers in rat lung at 28 days after administration. At the same time, Baicalin also diminished the level of interleukin-1beta (IL-1beta, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) in lung tissues. The protein expression of collagen I (Col-1), alpha-smooth muscle actin (alpha-SMA) and vimentin were down-regulated while E-cadherin (E-cad) was increased in Baicalin-treated rats. In addition, the Toll Like Receptor 4 (TLR4)/ nuclear factor kappaB (NF-kappaB) pathway was enabled at 28 days after silica infusion, and the treatment of Baicalin diminished the expression of TLR4 and NF-?B in the lungs of rat with silicosis. These results suggested that Baicalin inhibited the pulmonary inflammatory and fibrosis in a rat model of silicosis, which could be attributed to inhibition of the TLR4/NF-kappaB pathway.

[Interventional effect of asiaticosdide on rats exposed to silica dust].

Objective: To investigate the effect of asiaticoside for fibrosis in lung tissues of rats exposed to silica and to explore its possible mechanism. Methods: 144 SD male rats were randomly divided into control group, model group, positive drug control group, asiaticoside high-dose group, medium-dose group and low-dose group, each group included 24 rats. Rats in the control group were perfused with 1.0 ml of normal saline, and the other groups were given 1.0 ml 50 mg/ml SiO(2) suspension. Gavage of herbal was given from the next day after model establishment, once a day. Rats in the positive drug control group were administration with 30 mg/kg tetrandrine and rats in the low-dose group, medium-dose group and high-dose group were given 20 mg/kg, 40 mg/kg and 60 mg/kg asiaticoside for fibrosis respectively. Rats in the control group and the model group were given 0.9% normal saline. The rats were sacrificed in on the 14th, 28th and 56th day after intragastric administration and collect the lung tissues to detect the content of hydroxyproline, TGF-ß(1) and IL-18, observe the pathological changes of the lung tissues by HE and Masson staining and determine the expressions of Col-I, a-SMA, TGF-ß in lung tissues by Western Blot. Results: On the 14th day, 28th day and 56th day after model establishment, the lung tissues of rats in the model group showed obvious inflammatory response and accumulation of collagen fibers, and the degree of inflammation and fibrosis increased with time. The intervention of asiaticoside could effectively inhibit the pathological changes of lung tissues. The contents of hydroxyproline, IL-18 and TGF-ß1 in lung tissues of model group were higher than those in the control group (P<0.05) , while the level of hydroxyproline, IL-18 and TGF-ß1 in asiaticoside groups were significantly decreased, and the difference was statistically signicant (P<0.05) . Compared with the control group, the expression levels of Col-I, TGF-ß1and α-SMA in lung tissue of model group were increased (P<0.05) , while the expression level of Col-I, TGF-ß1 and α-SMA were decreased after the intervention of asiaticoside, and the difference was statistically signicant (P<0.05) . Conclusion: Asiaticoside can inhibit the increase of Col-I, TGF-ß1 and α-SMA content in the SiO(2)-induced lung tissues of rats, reduce the release of TGF-ß1 and IL-18 inflammatory factors in lung tissue, and then inhibit the synthesis and deposition of extracellular matrix in rat lung tissue, and improve silicosis fibrosis.

Association between Depressive Symptoms and Supplemental intake of Calcium and Vitamin D in Older Adults.

OBJECTIVES: We explored the associations between depressive symptoms and supplemental calcium and vitamin D intake in older adults. DESIGN: This was a prospective cohort study. PARTICIPANTS: 8,527 older adults aged ≥60 years from Zhejiang Major Public Health Surveillance Program (ZPHS) without depressive symptoms at baseline survey. MEASUREMENTS: Participants were divided into non-supplementation, calcium (Ca), vitamin D, and calcium plus vitamin D (CaD) groups based on their supplemental intake during the past year. Depressive symptoms were assessed using the Patient Health Questionnaire-9 (PHQ-9). Binary logistic regression analyses were performed to examine the association between depressive symptoms and supplemental intake. RESULTS: When compared to the non-supplementation group, the Ca group exhibited a significant odds ratio (OR) of 0.731 (95% CI: 0.552-0.967, P=0.028). After adjusting for age, sex, and Ca food sources, the OR was even smaller for the CaD group (OR: 0.326; 95% CI: 0.119-0.889, P=0.029). Additionally, our results indicated that taking Ca supplements ≥4 days/week yielded a significant OR of 0.690 (95% CI: 0.492-0.968) after full adjustment. Taking CaD supplements ≥4 days/week yielded a significant OR of 0.282 (95% CI: 0.089-0.898) after adjusting for age, sex, and Ca food sources. CONCLUSIONS: Supplemental intake of Ca or CaD ≥4 days/week can decrease the risk of depressive symptoms in older adults, although CaD supplements may be more effective.

Pathophysiological characteristics and gene transcriptional profiling of bone microstructure in a low calcium diet fed laying hens.

Calcium depletion is a valuable non-invasive tool for studying skeletal system disorders. A low calcium diet (LCD) was used to examine the pathophysiological characteristics and molecular mechanisms of osteoporotic damage in caged laying hens. Sixty 64-wk-old laying hens were randomly housed in single-bird cages, and the cages were divided into 2 treatments: fed a regular calcium diet (RCD, 3.69%) or a LCD (1.56%) for 8 wk. The diet-induced changes of serum bone remodeling indicators, bone strength, microstructure of the distal femur, and the gene expression profiling of keel bone were measured. Compared to RCD hens, LCD hens had higher activity of serum alkaline phosphatase and tartrate resistant acid phosphatase but lower serum calcium concentrations with reduced tibial and femoral mass, width, and strength (P < 0.05). In addition, LCD hens had greater densities of osteoclasts, osteoblasts, connective tissue cells, and osteoid in the trabecular bone (P < 0.05). The transcriptome analysis revealed that 563 unigenes were differentially expressed in keel bone between LCD and RCD hens. The Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that these differentially expressed genes were mainly associated with the osteoporosis-related signaling pathways involved in the biological functions of the bone cellular and extracellular structural modulations. The real-time PCR analysis further confirmed that the LCD enhanced the mRNA expression of collagen type 1 alpha 2, integrin-binding sialoprotein and periostin, but inhibited sclerostin expression. These findings indicate that LCD hens have a higher bone turnover and micro-architectural damage compared to RCD hens. The results further evidence dietary supplement of calcium is a critical nutrient strategy for improving hen skeletal health.

Thermoluminescence glow curves and optical stimulated luminescence of undoped alpha-Al2O3 crystals.

The characteristics of thermoluminescence (TL) and optical stimulated luminescence (OSL) in undoped alpha-Al2O3 single crystals were studied. The TL glow curves of the crystal samples irradiated at various dose levels were measured by RisØ TL/OSL-DA-15B/C reader with U-340 or 7-59 filters at different heating rates. The glow peaks measured with U-340 at approximately 210 degrees C of the undoped alpha-Al2O3 can be well fitted by first-order kinetic equation whereas the glow peaks measured with 7-59 filters are a composite of two first-order glow peaks. It indicates that the TL glow curves are dependent upon the filter used in the reader that is related to the emission spectra of luminescence materials. The OSL were also measured and fitted by two exponential functions to get the luminescence intensities. The TL and OSL dose responses of the undoped alpha-Al2O3 crystal were obtained in the dose range of 0.12-248 Gy and fitted by the composite action dose-response function to get nonlinear characteristic parameters. The TL and OSL dose responses are linear-sublinear.

Thermoluminescence of MgSO4 doped with Eu and P impurities.

CaSO4:Eu, MgSO4:Eu and MgSO4:Eu,P phosphors have been prepared and their thermoluminescence (TL) characteristics were studied. A main glow peak due to Eu2+ ions is seen at approximately 146 degrees C and 440 nm and glow peaks at approximately 145 degrees C, approximately 190 degrees C, approximately 260 degrees C and approximately 360 degrees C for 590 nm and 625 nm wavelengths are identified as Eu3+ ion emissions in MgSO4:Eu. Emission spectra in MgSO4:Eu and the MgSO4:Eu,P show that the MgSO4:Eu3+ glow peak at 260 degrees C for 590 nm and 625 nm shifts to 280 degrees C with enhanced intensity while the Eu2+ ion glow peak at 146 degrees C remains but with reduced intensity. The main glow peak at approximately 146 degrees C and 440 nm from Eu2+ ions shows significant difference from the characteristic glow peaks of Eu3+ ions. It is observed that the wavelength of the Eu2+ ion glow peak is inversely proportional to the radius of the cation of the host sulphate in alkaline-earth sulphate phosphors. By contrast the wavelengths of the Eu3+ ion glow peaks remain unchanged in different sulphates. Besides, the glow curve at approximately 146 degrees C obtained using a conventional blue sensitive reader shows simply the first order kinetics. It is concluded that the luminescence centres and distribution of traps related to Eu2+ ions are different from that of Eu3+ ions in MgSO4:Eu and MgSO4:Eu,P phosphors.

Isoquinoline alkaloids from Thalictrum delavayi.

Two new protoberberine alkaloids, 2,3,9,10-dimethylenedioxy-8-oxoprotoberberine (1) and 2,3,9,10-dimethylenedioxy-1,8-dihydroxyprotoberberine (2), together with nine known isoquinoline-type alkaloids were isolated from the roots of Thalictrum delavayi. Their structures were elucidated by spectral methods. Among these compounds, pseudoprotopine showed competitive inhibition activity by DA receptor binding assay (D1) in vitro. The competitive inhibitions were 87.5% (10(-4) M) and 15.6% (10(-6) M), respectively.

[Effects of YMB injection on hemorheology in rats].

This study was designed to elucidate the mechanism of YMB Injection on anti-CAD therapy. Fufang Danshen Injection was used as positive control and saline was taken as negative control to study the effects of traditional medicine YMB Injection on hemorheological parameters in normal rats and hypostasis rats. The results showed YMB Injection had no effect on hemorheological parameters of normal rats, but it could effectively decrease whole blood viscosity and whole blood reduction viscosity of hypostasis rats. The data implied that YMB Injection could decrease blood viscosity and hence improve blood supply of the graft in anti-CAD therapy.

[Effect of gandou decoction on copper metabolism of skin fibroblast of hepatolenticular degeneration model].

OBJECTIVE: To explore the cytological mechanism of Gandou Decoction (GDD) in treating hepatolenticular degeneration (HLD). METHODS: Twenty-three in vitro models of cultured skin fibroblast of HLD were established to observe the intracellular change of microelements (Cu2+, Zn2+, etc.) content before and after adding rabbit's serum containing GDD. RESULTS: After being treated with GDD contained serum for 24 hours, the intracellular content of Cu2+ decreased from (80.94 +/- 34.76) ng/mg to (46.90 +/- 22.14) ng/mg, P < 0.01, while that of Zn2+ increased from (140.43 +/- 33.81) ng/mg to (151.43 +/- 37.83) ng/mg, P < 0.01. CONCLUSION: GDD is quite effective in removing the intracellular copper and increasing the intracellular zinc.

Prevention by inhibitors of arachidonic acid cascade of liver carcinogenesis, cirrhosis and oxidative DNA damage caused by a choline-deficient, L-amino acid-defined diet in rats.

Effects of inhibitors of arachidonic acid (AA) cascade on the development of fatty liver, cirrhosis, glutathione S-transferase placental form (GST-P)-positive preneoplastic nodules, neoplastic nodules and generation of 8-hydroxydeoxyguanosine (8-OHdG), caused by a choline-deficient, L-amino acid-defined (CDAA) diet, were examined in Fischer 344 male rats by feeding CDAA diet supplemented with the inhibitors for 12 and 30 weeks. None of the inhibitors affected fatty liver. Among cyclooxygenase (COX) inhibitors, an irreversibly acting acetylsalicylic acid and a long-acting piroxicam, and to a much lesser extent the short-acting ibuprofen but not indomethacin, inhibited the development of cirrhosis, GST-P-positive and neoplastic nodules and generation of 8-OHdG. A phospholipase A2 inhibitor p-bromophenacylbromide (BPB) also exerted similar but lesser extent of inhibitory effects. Lipoxygenase inhibitors quercetin and nordihydroguiaretic acid inhibited GST-P-positive nodules but not cirrhosis or 8-OHdG. Present results suggest that perturbed AA cascade, particularly augmented COX pathway, might play key roles in the causation of liver lesions in the CDAA diet model.

[Determination of chemical constituents of the volatile oil from Curcuma longa by gas chromatography-mass spectrometry].

The study of chemical constituents of the volatile oil from Curcuma longa is reported. The volatile oil was extracted by steam distillation. Fifteen components in the oil were separated and identified by gas chromatography-mass spectrometry(GC/MS). The results were elucidated based on the NBS standard mass spectral data. The total ion current chromatogram showed their mass fraction by normalization method. alpha-curcumene, alpha-zingiberene, 1,8-cineole and zerumbone, which had been reported before were found in Curcuma longa volatile oil, but 1-(3-cyclopentylpropyl)-2,4-dimethylbenzene, beta-sesquiphellandrene, germacrene etc identified simultaneously in the oil had never been reported. The major chemical constituent of the volatile oil from Curcuma longa is alpha-curcumene.

[Effect of buyang huanwu decoction on platelet function and fibrinolytic activity of unstable angina pectoris patients].

OBJECTIVE: To explore the effect of Buyang Huanwu Decoction (BYHWD) on the platelet function and fibrinolytic activity of unstable angina pectoris patients. METHODS: Using randomized single blind method divided 60 unstable angina pectoris patients into conventional treatment (control) group and conventional plus BYHWD (TCM) group. RESULTS: BYHWD could lower the levels of platelet aggregation, thromboxane B2 and the inhibitor of fibrinolysinogen activator (P < 0.05), but the activity of histo-type of fibrinolysinogen activator was enhanced. CONCLUSION: BYHWD was effective in treating unstable angina pectoris.

Inhibition by acetylsalicylic acid, a cyclo-oxygenase inhibitor, and p-bromophenacylbromide, a phospholipase A2 inhibitor, of both cirrhosis and enzyme-altered nodules caused by a choline-deficient, L-amino acid-defined diet in rats.

Effects of inhibitors of arachidonic acid (AA) metabolism on the development of fatty liver, cirrhosis, glutathione-S-transferase placental form (GST-P)-positive nodules and the generation of 8-hydroxydeoxyguanosine (8-OHdG) and thiobarbituric acid-reactive substances (TBARS), caused by a choline-deficient, L-amino acid-defined (CDAA) diet, were examined in male Fischer 344 rats by feeding CDAA diets supplemented with the inhibitors for 12 and 30 weeks. Acetylsalicylic acid (ASA) (at doses of 0.1 and 0.2%) and p-bromophenacylbromide (BPB) (0.1 and 0.2%) were used as inhibitors of, respectively, cyclo-oxygenase and phospholipase A2, and quercetin (QU) (0.75 and 1.5%) and nordihydroguaiaretic acid (NDGA) (0.1 and 0.2%) as inhibitors of lipoxygenase. None of the inhibitors affected the development of fatty liver caused by the CDAA diet. ASA at a doe of 0.2% almost completely prevented the appearance of cirrhosis, GST-P-positive nodules, 8-OHdG and TBARS in seven out of 11 (63.7%) rats. BPB at a dose of 0.2% also exerted inhibitory effects on all of these lesions but to a lesser extent than ASA. QU and NDGA exerted inhibitory effects limited to the GST-P-positive nodule case. The results indicate that a perturbed AA metabolism, particularly of the cyclo-oxygenase pathway, derived secondarily from depletion of labile methyl groups or phosphatidylcholine, might play key roles in the cirrhosis, hepatocarcinogenesis and oxidative stress caused by a CDAA diet. The results also indicated a possible involvement of the lipoxygenase pathway in hepatocarcinogenic processes.

Use of site-directed mutagenesis to identify the galactosyltransferase binding sites for UDP-galactose.

Site-directed mutagenesis was utilized to identify binding sites for UDP-galactose in galactosyltransferase (EC 2.4.1.22). Mutant cDNAs were generated by a procedure based on PCR, and the mutated enzymes were expressed in E.coli cells. The mutant enzymes were purified by Ni-NTA Sephadex, and the degree of purification was judged by SDS-PAGE. Purified mutant GTs, F305L, P306V, N307S, N308S, showed dramatic decreases in activities in comparison with the activity of the wild-type GT. Enzyme kinetic analysis revealed that the Km values of F305L, P306V, N307S and N308S for UDP-galactose were, respectively, 9-, 11-, 50- and 20-fold higher than the Km of wild-type GT, but the Km values for manganese were not significantly different from that of the wild-type GT. The quartet mutant F305L/P306V/N307S/N308S showed no activity. From the results of this study it is concluded that amino acids, Phe-305, Pro-306, Asn-307 and Asn-308, in GT are most probably involved in GT catalysis or are located close to the UDP-galactose binding region but are not involved in the binding of manganese.

Prevention by acetylsalicylic acid of liver cirrhosis and carcinogenesis as well as generations of 8-hydroxydeoxyguanosine and thiobarbituric acid-reactive substances caused by a choline-deficient, L-amino acid-defined diet in rats.

Effects of acetylsalicylic acid (ASA) (aspirin) on the pathogenesis of fatty liver, cirrhosis and hepatocarcinogenesis caused by a choline-deficient L-amino acid-defined (CDAA) diet were examined in male Fischer 344 rats fed a CDAA diet supplemented with 0, 0.1, 0.2, 0.4 or 0.8% ASA for 30 weeks. ASA at concentrations of > 0.2% prevented the development of both cirrhosis and preneoplastic and neoplastic nodules, but without any directly associated prevention of fatty changes. ASA also prevented hepatocyte proliferation and the generation of thiobarbituric acid-reactive substances and 8-hydroxydeoxyguanosine caused by feeding the CDAA diet, analyzed, respectively, after 1, 12 and 12 weeks. The results clearly indicate that the anti-inflammatory drug ASA, which is not a lipotropic factor, can prevent the pathogenesis of cirrhosis and hepatocarcinogenesis caused by a CDAA diet, which is possibly partly associated with the prevention of reactive oxygen species production.

Induction of 8-hydroxydeoxyguanosine but not initiation of carcinogenesis by redox enzyme modulations with or without menadione in rat liver.

Inducibility of oxidative stress in rat liver in vivo by menadione-associated redox cycling activation under redox enzyme modulating conditions was examined by monitoring hepatocyte injury and 8-hydroxydeoxyguanosine (8-OHdG) levels of liver DNA. In addition, the treatment-associated liver tumor initiating activity was assessed in terms of development of gamma-glutamyl-transpeptidase (GGT)- and glutathione S-transferase placental form (GST-P)-positive foci and hyperplastic nodules. With or without following menadione treatment (50 mg/kg, i.g.), redox enzyme modulations of increased cytochrome P450 reductase activity induced by phenobarbital (PB)-Na (100 mg/kg, i.p. for 5 days), inhibition of DT-diaphorase by dicumarol (25 mg/kg, i.p.) and depletion of glutathione by phorone (200 mg/kg, i.p.), with or without further supplement of iron EDTA-Na-Fe(III) (70 mg/kg, i.p.), caused both substantial hepatocyte necrosis and 8-OHdG production in Fischer 344 male rats. Subsequent feeding with a 0.05% PB diet for 64 weeks resulted in slightly increased development of GGT-positive foci but not GST-P positive lesions or hyperplastic nodules, suggesting a lack of tumor-initiating activity of the oxidative DNA damage associated with redox enzyme modulations with or without menadione.

[Changes in monoamine neurotransmitters in CSF during inhibition of defensive hypertensive reactions by inputs of deep peroneal nerve].

Our previous work has indicated that excitation of hypothalamic defence area (HDA) could lead to an increase in the release of central monoamine neurotransmitters, pressor and other defence responses. The present experiments showed that stimulation of deep peroneal nerve with a current of low frequency and low intensity could inhibit the pressor, abolish the increase of noradrenaline and its metabolite MHPG and attenuate that of 5-HIAA (a 5-HT metabolite), dopamine and its metabolites DOPAC and HVA in CSF. The data suggest that inputs of deep peroneal nerve may inhibit the increased release of central monoamine neurotransmitters, and then defence reactions; and inhibition of NA release in some nuclei of the brain is an important mechanism of inhibition of defence pressor by inputs of deep peroneal nerve.