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.

[Interventional effect of metformin on pulmonary inflammation and pulmonary fibrosis in silicotic rats].

Objective: To investigate the interventional effect of metformin on pulmonary inflammation and pulmonary fibrosis in silicotic rats. Methods: In April 2019, 48 Wistar male rats of SPF grade were randomly divided into negative control group, metformin control group, silicon dioxide (SiO2) model group, low, medium and high dose metformin intervention group according to the random number table method, 8 rats in each group. The SiO2 model group and the low, medium and high dose metformin intervention groups were given 1 ml 50 mg/ml of SiO2 by intratracheal instillation, the negative control group and the metformin control group were given 1 ml normal saline by intratracheal instillation. 24 hours later, the low, medium and high dose metformin intervention groups and the metformin control group were treated with 100, 200, 400 and 400 mg/kg metformin daily, the control and SiO2 model groups received normal saline daily. Then the rats were sacrificed at the 28th day after SiO2 exposure. The changes of rat body weight and pathological examination of rat lung tissue were observed, and the lung organ coefficient, the content of hydroxyproline (HYP) , the expression levels of inflammatory factors transforming growth factor beta1 (TGF-ß1) , tumor necrosis factor-alpha (TNF-α) , interleukin-1beta (IL-1ß) and the protein expression of E-cadherin (E-Cad) , Vimentin, α-SMA were detected. Results: Compared with the negative control group, SiO2 model group had a significant decrease in the body weight of rats (P<0.05) , lung organ coefficient, alveolitis and fibrosis scores, HYP content and the levels of TGF-ß1, TNF-α, IL-1ß were all significantly increased (P<0.05) . Compared with the SiO2 model group, the weights of the rats in the medium and high dose intervention group of metformin increased significantly (P<0.05) . And after intervention with different doses of metformin, the lung organ coefficient, alveolitis and fibrosis scores, HYP content and the levels of TGF-ß1, TNF-α and IL-1ß were significantly decreased (P<0.05) . Immunohistochemistry and Western blotting results showed that compared with the negative control group, the expression of E-Cad of the SiO2 model group was decreased, and the expression levels of Vimentin and α-SMA were significantly increased (P<0.05) . After metformin intervention, the expression of E-Cad was significantly increased, the expression levels of Vimentin and α-SMA were significantly decreased (P<0.05) . Conclusion: Metformin can reduce lung tissue inflammation and fibrosis in rats exposed to SiO2 dust, which may be related to reducing the expression of inflammatory factors in lung tissue and inhibiting the EMT process.

Rehmanniae Radix Preparata suppresses bone loss and increases bone strength through interfering with canonical Wnt/ß-catenin signaling pathway in OVX rats.

Rehmanniae Radix Preparata (RRP) improves bone quality in OVX rats through the regulation of bone homeostasis via increasing osteoblastogenesis and decreasing osteoclastogenesis, suggesting it has a potential for the development of new anti-osteoporotic drugs. INTRODUCTION: Determine the anti-osteoporotic effect of RRP in ovariectomized (OVX) rats and identify the signaling pathway involved in this process. METHODS: OVX rats were treated with RRP aqueous extract for 14 weeks. The serum levels of tartrate-resistant acid phosphatase (TRAP), receptor activator of nuclear factor kappa-Β ligand (RANKL), alkaline phosphatase (ALP), and osteoprotegerin (OPG) were determined by ELISA. Bone histopathological alterations were evaluated by H&E, Alizarin red S, and Safranin O staining. Bone mineral density (BMD) and bone microstructure in rat femurs and lumbar bones were determined by dual-energy X-ray absorptiometry and micro-computed tomography. Femoral bone strength was detected by a three-point bending assay. The expression of Phospho-glycogen synthase kinase 3 beta (p-GSK-3ß), GSK-3ß, Dickkopf-related protein 1 (DKK1), cathepsin K, OPG, RANKL, IGF-1, Runx2, ß-catenin, and p-ß-catenin was determined by western blot and/or immunohistochemical staining. RESULTS: Treatment of OVX rats with RRP aqueous extract rebuilt bone homeostasis demonstrated by increasing the levels of OPG as well as decreasing the levels of TRAP, RANKL, and ALP in serum. Furthermore, RRP treatment preserved BMD and mechanical strength by increasing cortical bone thickness and epiphyseal thickness as well as improving trabecular distribution in the femurs of OVX rats. In addition, RRP downregulated the expression of DKK1, sclerostin, RANKL, cathepsin K, and the ratio of p-ß-catenin to ß-catenin, along with upregulating the expression of IGF-1, ß-catenin, and Runx2 and the ratio of p-GSK-3ß to GSK-3ß in the tibias and femurs of OVX rats. Echinacoside, jionoside A1/A2, acetoside, isoacetoside, jionoside B1, and jionoside B2 were identified in the RRP aqueous extract. CONCLUSION: RRP attenuates bone loss and improves bone quality in OVX rats partly through its regulation of the canonical Wnt/ß-catenin signaling pathway, suggesting that RRP has the potential to provide a new source of anti-osteoporotic drugs.

Characterization of the Microbial Communities in the Ant Lion Euroleon coreanus (Okamoto) (Neuroptera: Myrmeleontidae).

Euroleon coreanus (Okamoto) is widely distributed in China, and the larval stage can be treated as traditional Chinese medicine. However, the host-bacterium relationship remains unexplored, as there is a lack of knowledge on the microbial community of ant lions. Hence, in the current study, we explored the microbial community of the larval ant lion E. coreanus using Illumina MiSeq sequencing. Results indicated that a total of 10 phyla, 126 genera, and 145 species were characterized from the second instars of E. coreanus, and most of the microbes were classified in the phylum Proteobacteria. Cronobacter muytjensii was the most abundant species characterized in the whole body and gut of E. coreanus, and the unclassified species in the genera Brevundimonas and Lactobacillus were relatively more abundant in the head and carcass. In addition, no Wolbachia-like bacteria were detected, whereas bacteria like Francisella tularensis subsp. Holarctica OSU18 and unclassified Rickettsiella were first identified in ant lion E. coreanus.

Heterologous expression of Escherichia coli fructose-1,6-bisphosphatase in Corynebacterium glutamicum and evaluating the effect on cell growth and L-lysine production.

Fructose-1,6-bisphosphatase (FBPase), which is mainly used to supply NADPH, has an important role in increasing L-lysine production by Corynebacterium glutamicum. However, C. glutamicum FBPase is negatively regulated at the metabolic level. Strains that overexpressed Escherichia coli fructose-1,6-bisphosphatase in C. glutamicum were constructed, and the effects of heterologous FBPase on cell growth and L-lysine production during growth on glucose, fructose, and sucrose were evaluated. The heterologous fructose-1,6-bisphosphatase is insensitive to fructose 1-phosphate and fructose 2,6-bisphosphate, whereas the homologous fructose-1,6-bisphosphatase is inhibited by fructose 1-phosphate and fructose 2,6-bisphosphate. The relative enzyme activity of heterologous fructose-1,6-bisphosphatase is 90.8% and 89.1% during supplement with 3 mM fructose 1-phosphate and fructose 2,6-bisphosphate, respectively. Phosphoenolpyruvate is an activator of heterologous fructose-1,6-bisphosphatase, whereas the homologous fructose-1,6-bisphosphatase is very sensitive to phosphoenolpyruvate. Overexpression of the heterologous fbp in wild-type C. glutamicum has no effect on L-lysine production, but fructose-1,6-bisphosphatase activities are increased 9- to 13-fold. Overexpression of the heterologous fructose-1,6-bisphosphatase increases L-lysine production in C. glutamicum lysC(T311I) by 57.3% on fructose, 48.7% on sucrose, and 43% on glucose. The dry cell weight (DCW) and maximal specific growth rate (µ) are increased by overexpression of heterologous fbp. A "funnel-cask" diagram is first proposed to explain the synergy between precursors supply and NADPH supply. These results lay a definite theoretical foundation for breeding high L-lysine producers via molecular target.

Hypoglycemic activity of a polyphenolic oligomer-rich extract of Cinnamomum parthenoxylon bark in normal and streptozotocin-induced diabetic rats.

Cinnamon bark has been reported to be effective in the alleviation of diabetes through its antioxidant and insulin-potentiating activities. The water-soluble polyphenolic oligomers found in cinnamon are thought to be responsible for this biological activity. In this study, the hypoglycemic activity of a polyphenolic oligomer-rich extract from the barks of Cinnamomum parthenoxylon (Jack) Nees was studied in normal, transiently hyperglycemic, and streptozotocin (STZ)-induced diabetic rats. Oral administration of the extract at doses of 100, 200, and 300 mg/kg body wt. caused significant changes in body weight loss and fasting blood glucose levels of normal rats. In STZ-induced diabetic rats, after administration of the extract at doses of 100, 200, and 300 mg/kg body wt. over 14 days, the blood glucose levels were decreased by 11.1%, 22.5%, and 38.7%, respectively, and the plasma insulin levels were significantly increased over pre-treatment levels. In an oral glucose tolerance test, the extract produced a significant decrease in glycemia 90 min after the glucose pulse. These results suggest that Cinnamomum parthenoxylon polyphenolic oligomer-rich extract could be potentially useful for post-prandial hyperglycemia treatment.

A medicinal extract of Scutellaria baicalensis and Acacia catechu acts as a dual inhibitor of cyclooxygenase and 5-lipoxygenase to reduce inflammation.

A mixed extract containing two naturally occurring flavonoids, baicalin from Scutellaria baicalensis and catechin from Acacia catechu, was tested for cyclooxygenase (COX) and 5-lipoxygenase (5-LOX) inhibition via enzyme, cellular, and in vivo models. The 50% inhibitory concentration for inhibition of both ovine COX-1 and COX-2 peroxidase enzyme activities was 15 microg/mL, while the mixed extract showed a value for potato 5-LOX enzyme activity of 25 microg/mL. Prostaglandin E2 generation was inhibited by the mixed extract in human osteosarcoma cells expressing COX-2, while leukotriene production was inhibited in both human cell lines, immortalized THP-1 monocyte and HT-29 colorectal adenocarcinoma. In an arachidonic acid-induced mouse ear swelling model, the extract decreased edema in a dose-dependent manner. When arachidonic acid was injected directly into the intra-articular space of mouse ankle joints, the mixed extract abated the swelling and restored function in a rotary drum walking model. These results suggest that this natural, flavonoid mixture acts via "dual inhibition" of COX and LOX enzymes to reduce production of pro-inflammatory eicosanoids and attenuate edema in an in vivo model of inflammation.

Imaging cerebral gene transcripts in live animals.

To circumvent the limitations of using postmortem brain in molecular assays, we used avidin-biotin binding to couple superparamagnetic iron oxide nanoparticles (SPIONs) (15-20 nm) to phosphorothioate-modified oligodeoxynucleotides (sODNs) with sequence complementary to c-fos and beta-actin mRNA (SPION-cfos and SPION-beta-actin, respectively) (14-22 nm). The Stern-Volmer constant for the complex of SPION and fluorescein isothiocyanate (FITC)-sODN is 3.1 x 10(6)/m. We studied the feasibility of using the conjugates for in vivo magnetic resonance imaging (MRI) to monitor gene transcription, and demonstrated that these complexes at 40 mug of Fe per kilogram of body weight were retained at least 1 d after intracerebroventricular infusion into the left ventricle of C57Black6 mice. SPION retention measured by MRI as T(2)* or R(2)* maps (R(2)* = 1/T(2)*) was compared with histology of iron oxide (Prussian blue) and FITC-labeled sODN. We observed significant reduction in magnetic resonance (MR) T(2)* signal in the right cortex and striatum; retention of SPION-cfos and SPION-beta-actin positively correlated with c-fos and beta-actin mRNA maps obtained from in situ hybridization. Histological examination showed that intracellular iron oxide and FITC-sODN correlated positively with in vivo MR signal reduction. Furthermore, in animals that were administered SPION-cfos and amphetamine (4 mg/kg, i.p.), retention was significantly elevated in the nucleus accumbens, striatum, and medial prefrontal cortex of the forebrain. Control groups that received SPION-cfos and saline or that received a SPION conjugate with a random-sequence probe and amphetamine showed no retention. These results demonstrated that SPION-sODN conjugates can detect active transcriptions of specific mRNA species in living animals with MRI.

Biodegradation of benzene and its derivatives by a psychrotolerant and moderately haloalkaliphilic Planococcus sp. strain ZD22.

The potential for biodegradation of aromatic hydrocarbons simultaneously at low temperatures and under saline and alkaline conditions is not well understood, but such biodegradation would be useful for remediation of polluted sites. A psychrotolerant, moderately haloalkaliphilic pure culture using benzene as a sole source of carbon and energy was isolated by selective enrichment from alkaline and saline soils in the vicinity of the Daqing oil field in China. An analysis of the 16S rDNA gene sequence and morphological and physiological characteristics showed that this strain is a member of the genus Planococcus, and it was designated as strain ZD22. Strain ZD22 could grow at temperatures between 2 and 36 degrees C (pH 7.5-11) and salt concentrations from 0.5 to 25%. Its optimal conditions for biodegradation of benzene were 20 degrees C (pH 9.5) and 10% salt concentration. Strain ZD22 not only utilized benzene, toluene, ethylbenzene and o-xylene, but also degraded chlorobenzene, bromobenzene, iodobenzene and fluorobenzene. The kinetic model of strain ZD22 for benzene was solved to obtain mumax=0.34 h-1, Ks=0.041 mM, n=1.21, Sm=10.2 mM. To our knowledge, this is the first report of biodegradation of benzene and its derivatives simultaneously under multiple extreme conditions.

[The effect of salvia miltiorrhiza and shengmai on inflammatory mediator and renal function of post-operative patients with obstructive jaundice].

OBJECTIVE: To investigate the effect of salvia miltiorrhiza and shengmai (SS) on inflammatory mediator and renal function in patients with obstructive jaundice (OJ). METHODS: This study included three groups: the OJ patients receiving SS for 6 days after operation (the SS group, n = 15); the OJ patients not receiving SS (the OJ group, n = 15); the patients with other hepato-biliary disorders (the Non-OJ group, n = 15). The levels of ET, LPs, TNF-alpha, IL-6, IL-8 and urine RBP, TFR, ALB were measured one day before operation and 1, 4, 7 days after operation. RESULTS: The SS group's LPs, TNF-alpha, IL-6, IL-8, ET, RBP, TFR and ALB levels were obviously lower on Day 7 after operation as compared with those on Day 1 after operation (P < 0.05). The urine RBP and ALB levels were significantly lower in the SS group than in the OJ group (P < 0.05). CONCLUSION: The post-operative use of salvia miltiorrhiza and shengmai for patients with obstructive jaundice may effectively decrease the postoperative levels of plasma LPs, ET, TNF-alpha, IL-6 and IL-8. At the same time, salvia miltiorrhiza and shengmai may protect renal function by inhibiting inflammatory mediator and improving blood dynamics.

Modified Aloe barbadensis polysaccharide with immunoregulatory activity.

Aloe barbadensis polysaccharide was partially digested with cellulase and further purified by dialysis, stepwise ethanol precipitation, and size exclusion chromatography. Crude modified Aloe polysaccharide (MAP) activated macrophage cells and stimulated fibroblast growth. Under the same conditions, native Aloe barbadensis gel had no effect on macrophage activation. MAP prevented ultraviolet B (UVB) irradiation-induced immune suppression as determined by contact hypersensitivity (CHS) response in C3H/HeN mice. This in vivo activity was correlated with the activity of MAP to inhibit UVB irradiation-induced tumor necrosis factor alpha (TNF-alpha) release from human epidermoid carcinoma cells (KB cells). MAP with an average molecular weight of 80,000 Dalton (Da) contained mannose, galactose, and glucose in a ratio of 40:1.4:1.0. MAP was likely a linear, highly acetylated molecule.

[Clinical effect of Xinqingning combined low dose continuous gastrointestinal dialysis in treating uremia].

OBJECTIVE: To investigate the clinical effect of Xinqingning (XQN), a preparation of rhubarb, combined with low dose continuous gastrointestinal dialysis in treating uremia. METHODS: Patients of uremia were divided into three groups at random, eighteen patients in the group A were treated with 1000 ml gastrointestinal dialysate (non-absorbed mannitol solution) orally 2-3 times a day, twenty patients in the group B treated with the same therapy as that in the group A, but combined with XQN 5-10 tablets 3 times per day and 19 patients in the group C treated by orally taken coated aldehyde oxystarch 5-10 g, 3 times a day. The therapeutic course for the three groups were 11 months. The changes in clinical manifestation, renal function, nutritional condition, and electrolytes before and after treatment were observed. RESULTS: After treatment, significant improvement was revealed in aspects of uremic symptoms, quality of life, nutritional condition, serum creatinine, urea nitrogen, serum phosphate, uric acid, CO2 combining power, creatine clearance, body weight and arm muscular circumference in the group A and B, as compared with those in the group C. In comparison of the group A and B, the group B showed a lower serum triglyceride and slower progression of chronic renal failure (CRF). All the criteria were not improved in the group C and with serum creatinine raised significantly. CONCLUSION: XQN combined low dose continuous gastrointestinal dialysis therapy was effective in definitely treating uremia, it provides a new therapeutic means of non-replacement therapy for CRF with uremia.

Pikuroside: a novel iridoid from Picrorhiza kurroa.

A new iridoid, pikuroside (1), was isolated from the roots of Picrorhiza kurroa, together with three known iridoids, picroside I (2), picroside II (3), and 6-feruloyl catalpol (4). The structure of 1 was established by interpretation and full assignments of NMR spectral data. Pikuroside (1) had no antiinflammatory activity, although the crude extract and picroside II (3) demonstrated moderate activity.

Drug leads from the Kallawaya herbalists of Bolivia. 1. Background, rationale, protocol and anti-HIV activity.

Aqueous, organic and alcoholic extracts of over 100 samples of 60 species of Kallawaya medicinal herbs representing 30 plant families were assayed to compare their toxicity and ability to protect MT-2 T-lymphoblastoid cells from the cytopathic effect of human immunodeficiency virus (HIV). The results are reported as a therapeutic index (TI) which was > 25 for eighteen species, including seven > 50 and one > 100. The anti-HIV activity resided primarily in the aqueous rather than in the organic extracts and was concentrated in plants used in ethnomedicine to treat lung and liver diseases.

PIP: Plants are a rich source of anti-viral substances. The National Cancer Institute therefore annually screens about 1500 species from Africa, Southeast Asia, and South and Central America, but not Bolivia, for anti-HIV activity. Several unique compounds with anti-HIV activity have emerged from the program. The Kallawaya Indians of Bolivia follow a medical tradition from the Tiahuanaco (400-1145), Mollo (1145-1435), Inca (1438-1532), Spanish (1532-1825), and Bolivian Republic (from 1825) which is only recently starting to be reported. They use approximately 900 of the more than 2000 medicinal plants found across Bolivia. Aqueous, organic, and alcoholic extracts of more than 100 samples of 60 species of Kallawaya medicinal herbs representing 30 plant families were assayed to compare their toxicity and ability to protect MT-2 T-lymphoblastoid cells from the cytopathic effects of HIV. The therapeutic index (TI) of sampled species is defined as the ratio of anti-HIV activity to toxic concentration. A TI of greater than 25 was chosen as the prerequisite for future bioassay-directed isolation of the active components as leads for potential new anti-HIV drugs. TI was greater than 25 for 18 species, including seven greater than 50 and one greater than 100. The anti-HIV activity resided mainly in the aqueous extracts and was concentrated in plants used in ethnomedicine to treat lung and liver diseases.

[The structure of glyuranolide, a new triterpene of Glycyrrhiza uralensis Fisch].

Using a medium pressure liquid chromatographic system, a new triterpene lactone named glyuranolide [3 beta, 22 alpha-dihydroxy-11-oxo-delta 12-oleanene-27 alpha-methoxy carbonyl-29-oic acid (29, 22 alpha-) lactone.] was isolated from the crude sapogenins of Glycyrrhiza uralensis Fisch. Its structure was elucidated by IR, UV, FAB-MS, and various NMR spectra (including NOE, BBD, INEPT, SR, COSY, NOESY etc.).