Steam Explosion-Assisted Extraction of Ergosterol and Polysaccharides from Flammulina velutipes (Golden Needle Mushroom) Root Waste.

In this work, steam explosion (SE) was applied to prompt the rapid extraction of ergosterol and polysaccharides from Flammulina velutipes root (FVR) waste. Ultrasound-assisted saponification extraction (UASE) followed by water extraction was used to prepare ergosterol and polysaccharides. The results indicated that SE destroyed the complicated structure of FVR and increased its internal porosity and surface roughness. SE caused the thermal degradation of FVR's structural components and increased the polysaccharide content 0.97-fold. As a result, the extraction yield and efficiency of ergosterol and polysaccharides were improved. The theoretical maximum extraction concentration (C∞) and diffusion coefficient (D) were increased by 34.10% and 78.04% (ergosterol) and 27.69% and 48.67% (polysaccharides), respectively. The extraction yields obtained within 20-30 min of extraction time exceeded those of untreated samples extracted after several hours. For polysaccharides, SE led to a significant reduction in the average molecular weight, increased the percentage of uronic acids and decreased the neutral sugar percentage. The monosaccharide composition was changed by SE, with an increase in the molar ratio of glucose of 64.06% and some reductions in those of other monosaccharides. This work provides an effective method for the processing of fungi waste and adds to its economic value, supporting its high-value utilization in healthcare products.

Triptolide protects human retinal pigment epithelial ARPE-19 cells against high glucose-induced cell injury by regulation of miR-29b/PTEN.

Oxidative stress and inflammation are necessary pathogenic factors contributing to the aetiology of diabetic retinopathy (DR). Triptolide (TPL) is derived from the traditional Chinese herb lei gong teng with anti-inflammatory, immunosuppressive and antitumor activities. This article was developed to examine the effect of TPL on DR. ARPE-19 cells were pre-treated with TPL and then stimulated by high glucose (HG). We found that TPL treatment enhanced cell viability, decreased apoptosis and ROS production in HG-treated RPE cells. MiR-29b was low-expressed in HG-treated cells, but TPL raised its expression. In addition, the protective activity of TPL towards ARPE-19 cells was attenuated when miR-29b was reduced. By utilising bioinformatics evaluation, PTEN was predicted as a downstream target of miR-29b. Also, TPL obstructed PI3K/AKT signalling pathways in HG-treated ARPE-19 Cells. Taken together, TPL secured ARPE-19 cells from HG-induced oxidative damage via regulating miR-29b/PTEN axis.

Anti-hepatitis B virus activity of lithospermic acid, a polyphenol from Salvia miltiorrhiza, in vitro and in vivo by autophagy regulation.

ETHNOPHARMACOLOGICAL RELEVANCE: Salvia miltiorrhiza (the roots of S. miltiorrhiza Bunge, Danshen in Chinese), a traditional Chinese medicine, has been clinically used to prevent and treat various diseases, such as cardiovascular and cerebrovascular diseases, diabetes, and hepatitis B, in China and some other Asian countries. Lithospermic acid (LA), a polyphenol derived from S. miltiorrhiza, has been reported to exhibit multiple pharmacological properties, such as anti-inflammatory, anti-HIV, and anti-carbon tetrachloride-induced liver injury activities. However, little is known about the anti-hepatitis B virus (HBV) activity of LA. AIM OF THE STUDY: The study was projected to investigate the anti-HBV activity of LA in vitro (HepG2.2.15 and pHBV1.3-transfected HepG2 cells) and in vivo (pAAV-HBV1.2 hydrodynamic injection [HBV-HDI] mice) and explore the potential mechanism as well. MATERIALS AND METHODS: Hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) contents were detected by ELISA kits. HBV DNA and hepatitis B core antigen (HBcAg) levels were evaluated by quantitative real-time polymerase chain reaction and immunohistochemistry assay, respectively. The proteins in autophagy process, lysosomal acidic function, and autophagy-related signaling pathways were examined by Western blot. Transmission electron microscopy was used to observe the number of autophagosomes and autolysosomes. Confocal microscopy was applied to analyze the autophagic flux and lysosomal acidification, using mCherry-enhanced green fluorescent protein (EGFP)-microtubule-associated protein light chain (LC)3 and lysosomal probes, respectively. RESULTS: LA exhibited anti-HBV activity by inhibiting HBV DNA replication in HepG2.2.15 and pHBV-transfected HepG2 cells in dose- and time-dependent manners and hampering HBsAg and HBeAg levels in HepG2.2.15 cells to a certain extent. LA reduced HBV DNA, HBsAg/HBeAg, and HBcAg levels in the serum/liver tissues of HBV-HDI C57BL/6 mice during the 3-week treatment and suppressed the withdrawal rebound of HBV DNA and HBsAg in the mice serum. LA increased LC3-II protein expression and the number of autolysosomes/autophagosomes and promoted the degradation of sequestosome 1(p62) protein in vitro and in vivo. LA enhanced the co-localization of LC3 protein with autolysosomes, further confirming the ability of LA to induce a complete autophagy. Knockdown of autophagy-related gene (Atg) 7 or 5 in vitro and administration of 3-methyladenine (an autophagic inhibitor) in vivo disabled the inhibitory efficacy of LA on HBV DNA replication, suggesting that the anti-HBV efficacy of LA depended on its ability of inducing autophagy. LA could enhance lysosomal acidification and improve the function of lysosomes by promoting the protein expression of lysosomal-associated membrane protein (LAMP)-1, LAMP-2, and mature cathepsin D, which may contribute to the autophagic induction of LA. LA inhibited the activation of AKT and mammalian target of rapamycin (mTOR) induced by HBV, which was reversed by IGF-1 (an agonist of the PI3K/AKT/mTOR signaling pathway), indicating that LA elicited autophagy through hampering the PI3K/AKT/mTOR signaling pathway. CONCLUSION: We revealed the anti-HBV activity and mechanism of LA in vitro and in vivo. This study facilitates a new understanding of the anti-HBV potent components of S. miltiorrhiza and sheds light on LA for further development as an active constituent or candidate used in the therapy against HBV infection.

Efficacy of epalrestat combined with alprostadil for diabetic nephropathy and its impacts on renal fibrosis and related factors of inflammation and oxidative stress.

OBJECTIVE: To explore the efficacy of epalrestat (Ep) combined with alprostadil (Alp) in the treatment of diabetic nephropathy (DN) and its impacts on renal fibrosis (RF) and inflammation and oxidative stress (OS)-related factors. METHODS: In this retrospective study, 120 patients with DN treated in the Cangzhou Central Hospital from January 2020 to January 2021 were selected as the research subjects. Among them, 80 cases treated with Ep combined with Alp were assigned to group A, and the rest 40 patients treated with Alp only were assigned to group B. The two groups were compared with respect to the following items: serum OS indexes (malondialdehyde, MDA; superoxide dismutase, SOD; total antioxidant capacity, TAOC), inflammatory factors (tumor necrosis factor-α, TNF-α; interleukin-2, IL-2), RF index transforming growth factor-ß1 (TGF-ß1), urinary protein indexes (urinary albumin excretion, UAE; serum albumin, ALB), blood glucose (fasting blood glucose, FBG), fasting C-peptide, postprandial 2hC peptide levels, overall response rate (ORR) and incidence of adverse reactions. RESULTS: Compared with group B, the levels of MDA, TNF-α, IL-2 and TGF-ß1 were lower, while SOD and TAOC were higher in group A. In addition, ALB was higher, while UAE and FBG were lower in group A as compared with group B. Moreover, group A had a higher ORR and fewer adverse reactions as compared with group B. CONCLUSION: The combined therapy of Ep and Alp is more effective in the treatment of DN. This combination can effectively reduce RF and better alleviate inflammation and OS.

CPSF1 mediates retinal vascular dysfunction in diabetes mellitus via the MAPK/ERK pathway.

This study investigated the expression and underlying molecular mechanism of CPSF1 in diabetic retinopathy. Streptozotocin (STZ)-induced Sprague-Dawley (SD) rats were employed as a diabetic model, and high-glucose (HG)-induced human retinal vascular endothelial cells （HRVECs）were used as an in vitro experimental model to explore the effect of CPSF1. The results showed that CPSF1 was downregulated in diabetic retinopathy (DR) tissues and HRVECs under HG conditions. Adeno-associated viral CPSF1 attenuated histological abnormalities of retinas. CPSF1 regulates the apoptosis, migration, and vascularisation of HRVECs under HG conditions in vitro. CPSF1 mediates retinal vascular dysfunction by suppressing the phosphorylation mechanism in the mitogen-activated protein kinase/extracellular-signal-regulated kinase (MAPK/ERK) pathway in DR. In conclusion, CPSF1 may be associated with the development of DR, and upregulated CPSF1 alleviates apoptosis and migration via MAPK/ERK pathway.

miR-340 Promotes Retinoblastoma Cell Proliferation, Migration and Invasion Through Targeting WIF1.

BACKGROUND: MicroRNAs (miRNAs) function as important regulators of gene expression involved in tumor pathogenesis, including retinoblastoma. However, the expression profiles and potential roles in retinoblastoma are still largely unclear. MATERIAL AND METHODS: Differentially expressed miRNAs (DEmiRs) and genes (DEGs) in retinoblastoma were extracted from Gene Expression Omnibus (GEO) repository. Expression levels of miR-340 and WIF1 were detected in retinoblastoma tissues and cell lines by qRT-PCR. Both gain-of-function and loss-of-function experiments were performed to explore the effects of miR-340 on cell proliferation, migration and invasion. Bioinformatics analysis and luciferase reporter assay were used to explore the interaction between miR-340 and WIF1. RESULTS: A total of 11 DEmiRs were identified in retinoblastoma tissue and blood samples. Among them, we validated that miR-340 was the most highly expressed miRNA and correlated with tumor size, ICRB stage and optic nerve invasion. miR-340 was observed to enhance the proliferation, migration and invasion capacity of retinoblastoma cells. We then identified 26 DEGs from 3 retinoblastoma GEO datasets and subsequently constructed a miRNA-mRNA regulatory network. Further analysis revealed that WIF1 was a direct target of miR-340. Moreover, overexpression of WIF1 could repress retinoblastoma progression induced by miR-340 in vitro and in vivo. CONCLUSION: Collectively, miR-340 functioned as an oncomiRNA to promote retinoblastoma cell proliferation, migration and invasion via regulating WIF1. Our data also provided multiple miRNAs and genes that may contribute to a better understanding of retinoblastoma pathogenesis.

Effects of daphnetin on lipid metabolism, insulin resistance and oxidative stress in OA­treated HepG2 cells.

Non­alcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease, and has high rates of morbidity and mortality worldwide. Daphnetin (DAP) possesses notable antioxidative, anti­inflammatory and anticoagulant activities; DAP is an active ingredient extracted from Daphne Koreana Nakai. To investigate the effects and the underlying mechanism of DAP on NAFLD, we treated HepG2 cells with oleic acid (OA) and DAP simultaneously and non­simultaneously. In the simultaneous treatment condition, HepG2 cells were co­treated with 0.5 mM OA and DAP (5, 20, and 50 µM) for 24 h. In the non­simultaneous treatment conditions, HepG2 cells were pretreated with 0.5 mM OA for 24 h, and then treated with DAP (5, 20 and 50 µM) for 24 h. Following the aforementioned treatments, the biochemical indexes associated with NAFLD were measured as follows: i) The intracellular contents of triglyceride (TG), reactive oxygen species (ROS) and fluorescent glucose 2­[N­(7­nitrobenz­2­oxa­1,3­diazol­4­yl) amino]­2­deoxyglucose were analyzed with corresponding detection kits; and ii) the cellular expression levels of glycolipid metabolism­ and oxidative stress­related genes, including 5'AMP­activated protein kinase (AMPK), sterol regulatory element­binding protein­1C (SREBP­1C), patatin­like phospholipase domain­containing protein 3 (PNPLA3), peroxisome proliferator­activated receptor α (PPARα), phosphoinositide 3­kinase (PI3K), protein kinase B (AKT), nuclear factor­like 2 (Nrf2), cytochrome P450 (CYP) 2E1 and CYP4A were determined by reverse transcription­quantitative polymerase chain reaction and western blotting. The results revealed the potential mechanism underlying the effects of DAP on NAFLD in vitro: i) By increasing the phosphorylation of AMPK, DAP inhibited the expression of SREBP­1C and PNPLA3, and induced that of PPARα. Lipid accumulation within hepatocytes was reduced; ii) by upregulating PI3K expression and pAKT/AKT levels, DAP may alleviate insulin resistance and promote hepatocellular glucose uptake; and iii) by upregulating the expression of Nrf2, DAP downregulated the expression of CYP2E1 and CYP4A, and the levels of reactive oxygen species in hepatocytes.

Protocatechuic acid inhibits hepatitis B virus replication by activating ERK1/2 pathway and down-regulating HNF4α and HNF1α in vitro.

AIMS: Protocatechuic acid (PCA) is a phenolic compound found in many antiviral Chinese herbal medicines. HNF4α and HNF1α, the members of hepatocyte nuclear factor (HNF) family, play an important regulatory role in the gene transcription of hepatitis B virus (HBV). Previous studies found that PCA inhibited HBV antigen secretion and HBV DNA replication in HepG2.2.15 cells, but its anti-HBV mechanism has not been fully understood. We aim to illustrate the anti-HBV mechanism of PCA. MATERIALS AND METHODS: MTT was used to estimate cytotoxicity. The content of HBsAg or HBeAg was detected using an enzyme-linked immunosorbent assay kit. HBV DNA in cell-free culture media was detected by PCR kit. HNF1α and HNF4α mRNA expression was detected by real-time PCR. HNF1α, HNF4α and ERK1/2 protein expression was detected by western blotting and HBV promoter activity was tested by luciferase reporter assay. KEY FINDINGS: Our results demonstrated that PCA inhibited the gene transcription and protein translation of HNF1α and HNF4α in Huh7 and HepG2.2.15 cells, as well as the promoter activities of HBV X and preS1 in Huh7 cells transfected with the luciferase reporter plasmid of HBV promoter. Further study suggested that PCA induced the phosphorylation of extracellular-signal-related kinase (ERK) 1/2, and thereby inhibited HNF4α and HNF1α expression in HepG2.2.15 cells to exert its antiviral activity. SIGNIFICANCE: To our knowledge, this study is the first to reveal the anti-HBV mechanism of PCA. Our results demonstrate that PCA inhibits HBV replication by activating ERK1/2 pathway and subsequently down-regulating HNF4α and HNF1α in HepG2.2.15 cells.

[Contrast sensitivity analysis about fundus changes of hypertensive disorders complicating pregnancy].

OBJECTIVE: To observe corrected visual acuity and contrast sensitivity (CS) in patients with hypertensive disorders complicating pregnancy accompanied by fundus changes.
 Methods: Ninety-eight patients with hypertensive disorders complicating pregnancy and 50 healthy pregnant women underwent eye examination, including corrected visual acuity and fundus examination, and CS. Differences in corrected visual acuity and contrast sensitivity between the 2 groups were analyzed with two independent samples t-test analysis, while correlation between vision and contrast sensitivity in patients was evaluated by using spearman correlation test. Difference in CS was compared between the early and advanced stage of fundus diseases.
 Results: Corrected visual acuity and contrast sensitivity in patient with hypertensive disorders complicating pregnancy were lower than that in the control group (P<0.01). Corrected visual acuity in patients was associated with contrast sensitivity at variously spatial frequencies (P<0.01), showing the most correlation in contrast sensitivity at 6 of spatial frequency (r=0.87). Compared with the early stage, the CS in the advanced patients with fundus diseases was decreased (P<0.01).
 Conclusion: The visual acuity and contrast sensitivity in patient with hypertensive disorders complicating pregnancy are reduced. The CS (6.0 c/d) has the largest correlation with corrected visual acuity. Comparing with the visual acuity, contrast sensitivity can be more comprehensive in evaluation of retinal function damage in patients with hypertensive disorders complicating pregnancy.

Effects of 6-Month Folic Acid Supplementation on Cognitive Function and Blood Biomarkers in Mild Cognitive Impairment: A Randomized Controlled Trial in China.

BACKGROUND: This study is to examine the effects of folic acid supplementation on cognitive function in Chinese older adults with mild cognitive impairment who are unexposed to folic acid fortification and assess cognitive functioning in relation to folate, homocysteine, and vitamin B12 values at baseline. METHODS: This was a single-center, randomized, controlled trial in Tianjin, China; 180 individuals aged 65 years and older who had mild cognitive impairment were assigned randomly to one of two groups: (a) those treated with oral folic acid (400 µg/day) and (b) those treated via conventional treatment. Tests of cognitive performance and biomarkers were measured at baseline, 3 months, and 6 months. Analysis was by intention-to-treat. Changes in cognitive or clinical function were analyzed by repeated-measure analysis of variance or mixed-effects models. This trial has been registered with the trial number ChiCTR-TRC-13003227. RESULTS: Total of 159 participants (intervention group: 80; control group: 79) completed the trial. Repeated-measure analysis of variance showed significant improvements in serum folate (ηp (2) = 0.712, p = .009), homocysteine (ηp (2) = 0.119, p = .017), serum vitamin B12 (ηp (2) = 0.144, p = .022), and S-adenosylmethionine (ηp (2) = 0.117, p = .033) in the intervention group over the control group. Folic acid supplementation improved Full Scale IQ (p = .031; effect size d = 0.168), Digit Span (p = .009; d = 0.176), and Block Design (p = .036; effect size d = 0.146) scores at 6 months in comparison to the control. There were no significant findings for all other cognitive measures. CONCLUSION: There was a beneficial effect from relatively short-term folate supplementation on cognitive functioning in later life. Larger-scale, randomized, controlled trials of longer duration in selected age groups are needed.

[Experimental study of glyceraldehyde cross-linking of posterior scleral on FDM in guinea pigs].

OBJECTIVE: The purpose of this study is to observe the effects of glyceraldehyde cross-linking on sclera biomechanical strength and experimental myopia. METHODS: 50 three weeks aged guinea pigs were randomly divided into 5 groups: A, B, C, D, E. Each group had 10 guinea pigs. The right eye was set as the experimental eye, the left eye was used as control. Group A: 7 days mask; Group B: 21 days mask, plus physiological saline retrobulbar injection at mask day 1, 8, 15; Group C: 21 days mask, plus 0.05 mol/L glyceraldehyde retrobulbar injection at mask day 1, 8, 15; Group D: 21 days mask, plus 0.5 mol/L glyceraldehyde retrobulbar injection at mask day 1, 8, 15; Group E: normal control group. Several parameters of 7 guinea pigs of each group were measured before and after deprivation (mask day 7, 14, 21), including ocular axial length, refractive error, ultimate stress (σmax) (MPa), ultimate strain(εmax) (%) and 6% elastic modulus (MPa). The effects of glyceraldehyde on adjacent tissues of the rest 3 guinea pigs were detected by histopathological and immunohistochemical studies. Differences between experiment eyes and contralateral, eyes were compared with paired t test and correlative analysis. RESULTS: The experimental eyes appeared the increase of the the vitreous cavity length, the axial length and myopia after deprivation. In group A, group B and group C, the differences of the length of the vitreous cavity (group A (2.991 ± 0.078) mm, group B (2.961 ± 0.038) mm and group C (2.936 ± 0.021) mm), the axial length(group A (7.263 ± 0.133) mm, group B (7.732 ± 0.099) mm and group C (7.665 ± 0.055) mm) and refractive error (group A (-2.214 ± 2.881) D, group B (-4.525 ± 2.415) D and group C (-1.607 ± 0.866) D) between the experimental eye and the fellow eye was statistically significant (vitreous cavity = 3.234, 4.758, 7.608; Pvitreous cavity = 0.018, 0.002, 0.001; axial = 3.198,, 4.758, 7.608; Paxial = 0.019, 0.002, 0.000; refraction = -7.120, -4.020, -6.334; Prefraction = 0.000, 0.005, 0.001). In group D and group E, there is no difference between deprived eye and control eye about the length of the vitreous chamber as well as axial length (vitreous = 0.542, -0.646; Pvitreous cavity = 0.607, 0.539; axial = 0.542, -0.646; Paxial = 0.607, 0.539). The experimental eye ((-3.921 ± 0.874)D) and the fellow eye ((-3.321 ± 1.205)D) of group D, the difference of diopter was statistically significant (refraction = -3.154, Prefraction = 0.020). At the end of the experiment, the change of diopter of experimental eye of group B, C, D, E was significantly different (F = 61.249, P = 0.000). The difference of diopter change between group B ((8.800 ± 0.616) D), group C ( (7.236 ± 2.198) D), group D ( (6.271 ± 1.112) D) and the normal control group ((0.934 ± 0.158) D) was statistically significant (PB = 0.000, PC = 0.000, PD = 0.000). At the end of the experiment, the ultimate stress and 6% elastic modulus of group B experimental eye was (7.988 ± 3.677) MPa (P = 0.002) and (19.938 ± 4.871) MPa (P = 0.001), decreased 10.06% and 34.17% respectively. On the other hand the ultimate strain was (28.6 ± 3.6) % (P = 0.034), increased 19.17%. After the cross-linking treatment, the ultimate stress and 6% elastic modulus of group C experimental eye was (9.244 ± 0.806) MPa (P = 0.001) and (26.180 ± 4.388) MPa (P = 0.031) , decreased 23.13% and 13.34%, the ultimate strain was (26.2 ± 1.0) % (P = 0.016) , increased 12.93% separately. The ultimate stress of group D experimental eye was (12.476 ± 2.507) MPa (P = 0.580), decreased 5.50%, 6% elastic modulus was (30.446 ± 3.410) MPa (P = 0.314), increased 6.53%, ultimate strain was (23.8 ± 1.8) % (P = 0.253), decreased 4.42% respectively. Ultrastructure examination showed that, decreased scleral thickness with fibers lined up in order, without inflammatory cells Infiltration. Expressions of matrix metalloproteinases-2(MMP-2) mainly decrease in the episcleral tissue. The stroma of choroid, and the outer plexiform layer. CONCLUSIONS: Glyceraldehyde is a safe and effective cross-linking agent that could significantly enhance the sclera biomechanical strength. Glyceraldehyde cross-linking method could effectively control the development of myopia in animal model.

In vivo effect of triptolide combined with glycyrrhetinic acid on rat cytochrome P450 enzymes.

Triptolide (TP) is a major active component in Tripterygium root, but its therapeutic window was very narrow due to its severe multi-organ toxicity. In this work, the effect of TP combined with glycyrrhetic acid (GA) on mRNA expression and activity of four cytochrome P450 (CYP) enzymes in rat liver was studied after intragastric administration of TP (0.05, 0.3 and 0.6 mg x kg(-1) x day(-1)) and TP (0.6 mg x kg(-1) x day(-1)) combined with GA (30 mg x kg(-1) x day(-1)) for 7 consecutive days. Compared with the control, the high dose of TP significantly up-regulated the mRNA expression levels of CYP2E1, 1A2, 3A1 and 2C11, the co-administration of TP and GA further up-regulated the mRNA expression levels of CYP3A1, 2C11 and 2E1 as compared with the high dose of TP. Meanwhile, TP at high dose and combined with GA significantly increased CYP3A-associated testosterone 6beta-hydroxylation activity (2.2-fold and 4.1-fold, respectively) as compared with the control. Because TP is mainly metabolized by CYP3A2 in male rats, the present work indicated that TP-induced increase of CYP3A activity might be an important reason for the rapidly metabolic clearance of TP in rat liver, and GA can reduce the hepatotoxicity of TP by promoting its hepatic metabolic clearance. Furthermore, the results also suggest that the drug interactions might be occurred when TP and GA were co-administered with other CYP3A substrate drug.

Effects of brucine combined with glycyrrhetinic acid or liquiritin on rat hepatic cytochrome P450 activities in vivo.

Abstract: The activities of four CYP450 enzymes (CYP3A, 1A2, 2El and 2C) and the mRNA expression levels of CYP1A2, 2El, 2Cll and 3A1 in rat liver were determined after Wistar rats were orally administered with brucine (BR) at three dosage levels (3, 15 and 60 mg.kg-1 per day) and the high dose of BR combined with glycyrrhetinic acid (GA, 25 mg.kg-1 per day) or liquiritin (LQ, 20 mg.kg-1 per day) for 7 consecutive days. Compared with the control, brucine caused 24.5% and 34.6% decrease of CYP3A-associated testosterone 6beta-hydroxylation (6betaTesto-OH) and CYP2C-associated tolbutamide hydroxylation (Tol-OH), respectively, and 146.1% increase of CYP2El-associated para-nitrophenol hydroxylation (PNP-OH) at the high dose level. On the other hand, (BR+GA) caused 51.4% and 33.5% decrease, respectively, of CYP2El-associated PNP-OH and CYP1A2-associated ethoxyresorufin-O-de-ethylation (EROD) as compared with the high dose of BR group. Meanwhile, (BR+LQ) caused 41.1% decrease of CYP2El-associated PNP-OH and 37.7% increase of CYP2C-associated Tol-OH. The results indicated that the co-administration of BR with GA or LQ had effect on mRNA expression and activities of the CYP450 enzymes mentioned above to some extent, and the in vivo antagonism of LQ on BR-induced CYPs adverse effects and the in vivo inhibitory action of GA on CYP2E1 and 1A2 might play an important role in the detoxification of Radix Glycyrrhizae against Strychnos nux-vomica L.

[Pharmacokinetics of gastrodin from compound Tianma granule in rats].

To study the influence of the compatibility of ophiopogonis tuber and Chinese magnoliavine fruit with gastrodia rhizome on the pharmacokinetics of gastrodin in rat, three dosages of compound Tianma granule extract (equivalent to gastrodin 50, 100, 200 mg x kg(-1)) and one dosage of Tianma extract (equivalent to gastrodin 100 mg x kg(-1) were administered to rats by intragastric administration separately. Plasma samples were collected at different times and treated with methanol and acetonitrile to precipitate protein. The contents of gastrodin in plasma were determined by HPLC method. The mean plasma concentration-time curves of different medication administration teams were processed with WinNonlin 5.2.1 pharmacokinetic software. The pharmacokinetic parameters of different medication administration teams were analyzed with SPSS statistics 17.0 software. The results indicated that the in vivo kinetic process of gastrodin was fitted to first-order absorption un-compartment model at low, middle dosages and zero-order absorption un-compartment model at high dosage of compound Tianma granule extract. By comparison with the pharmacokinetic parameters of gastrodin (100 mg x kg(-1)) in Tianma extract, the significant decrease for Cmax and significant increase for MRT0-infinity in compound Tianma granule extract indicated that the compatibility of ophiopogonis tuber and Chinese magnoliavine fruit with Gastrodia rhizome can delay the absorption, reduce the elimination rate and prolong the action time of gastrodin in vivo.

New modified iris suture technique for pupillary dilation in aphakic eyes during vitreoretinal surgery.

To describe a modified simple iris suture for pupillary dilation technique during vitrectomy in cases with a miotic pupil. Four translimbal incisions were created with a sharp straight blade at 1:30, 10:30, 4:30, and 7:30 o'clock, respectively. The straight needle of 10-0 polypropylene suture and a Sinskey IOL hook was used to displace the pupillary margin toward the limbus. In 3 cases, four sutures caused a 6-mm to 9-mm square-shaped pupil, and the pupil was allowed to return to a smaller size at the end of the operation. It is simple and may reduce postoperative complications.

[Pharmacokinetics of puerarin and hydrochlorothiazide from maijunan tablets in rats].

After oral administration of low, middle, high dose of simulative Maijunan tablets to SD rats and puerarin, hydrochlorothiazide at middle dosage to SD rats separately, plasma samples were collected at different times and treated with acetonitrile to precipitate protein. The contents of puerarin and hydrochlorothiazide in plasma were determined by HPLC method. The mean plasma concentration-time profiles of puerarin and hydrochlorothiazide at different dosages of medication administration were processed by 3P97 pharmacokinetic software and SPSS statistics 17.0 software. The results indicated that the in vivo kinetic processes of puerarin in rats were all fitted to a two-compartment open model and hydrochlorothiazide fitted to a one-compartment open model. Hydrochlorothiazide in vivo kinetic process in rats was in accordance with the linear dynamics. The combination of hydrochlorothiazide and rhynchophylla with pueraria promoted the absorption, reduced the elimination rate and prolonged the action time of puerarin in vivo. Meanwhile, the combination also promoted the absorption rate and the bioavailability, prolonged the action time and the accumulation time of hydrochlorothiazide in vivo.

[Study on the interaction between cinnamic acid and human serum albumin by fluorescence quenching method].

The albumin is the richest protein in blood circulatory system, which can combine with many drugs and play an important role in transporing protein. In the present work, the non-covalent interaction between human serum albumin and cinnamic acid was studied by using fluorescence quenching method. The results showed that cinnamic acid had a powerful ability to quench the fluorescence of human serum albumin at excitation and emission wavelengths of lambda(ex) = 286 nm and lambda(ex) = 340 nm, respectively, in the reaction solution of pH 7.4. The binding constants(K) at 37 and 47 degrees C were found to be 1.276 7 x 10(3) and 3.404 1 x 10(3) L x mol(-1), with the number of binding site(n) of 0.758 6 and 0.835 6, respectively, suggesting that the reaction temperature is advantageous for the binding reaction in a way. The changes in the thermodynamic parameters of binding interaction at 37 and 47 degrees C indicated that the main binding force between cinnamic acid and human serum albumin was hydrophobic force. These provide important information for studying the pharmacological effects of cinnamic acid and the influence of cinnamic acid on the configuration change of HSA.

LC/MS/MS for identification of in vivo and in vitro metabolites of jatrorrhizine.

The in vivo and in vitro metabolism of jatrorrhizine has been investigated using a specific and sensitive LC/MS/MS method. In vivo samples including rat feces, urine and plasma collected separately after dosing healthy rats with jatrorrhizine (34 mg/kg) orally, along with in vitro samples prepared by incubating jatrorrhizine with rat intestinal flora and liver microsome, respectively, were purified using a C(18) solid-phase extraction cartridge. The purified samples were then separated with a reversed-phase C(18) column with methanol-formic acid aqueous solution (70:30, v/v, pH3.5) as mobile phase and detected by on-line MS/MS. The structural elucidation of the metabolites was performed by comparing their molecular weights and product ions with those of the parent drug. As a result, seven new metabolites were found in rat urine, 13 metabolites were detected in rat feces, 11 metabolites were detected in rat plasma, 17 metabolites were identified in intestinal flora incubation solution and nine metabolites were detected in liver microsome incubation solution. The main biotransformation reactions of jatrorrhizine were the hydroxylation reaction, the methylation reaction, the demethylation reaction and the dehydrogenation reaction of parent drug and its relative metabolites. All the results were reported for the first time, except for some of the metabolites in rat urine.

Liquid chromatography-electrospray ionization ion trap mass spectrometry for analysis of in vivo and in vitro metabolites of scopolamine in rats.

In vivo and in vitro metabolism of scopolamine is investigated using a highly specific and sensitive liquid chromatography-mass spectrometry (LC-MSn) method. Feces, urine, and plasma samples are collected individually after ingestion of 55 mg/kg scopolamine by healthy rats. Rat feces and urine samples are cleaned up by a liquid-liquid extraction and a solid-phase extraction procedure (C18 cartridges), respectively. Methanol is added to rat plasma samples to precipitate plasma proteins. Scopolamine is incubated with homogenized liver and intestinal flora of rats in vitro, respectively. The metabolites in the incubating solution are extracted with ethyl acetate. Then these pretreated samples are injected into a reversed-phase C18 column with mobile phase of methanol-ammonium acetate (2 mM, adjusted to pH 3.5 with formic acid) (70:30, v/v) and detected by an on-line MSn system. Identification and structural elucidation of the metabolites are performed by comparing their changes in molecular masses (DeltaM), retention-times and full scan MSn spectra with those of the parent drug. The results reveal that at least 8 metabolites (norscopine, scopine, tropic acid, aponorscopolamine, aposcopolamine, norscopolamine, hydroxyscopolamine, and hydroxyscopolamine N-oxide) and the parent drug exist in feces after administering 55 mg/kg scopolamine to healthy rats. Three new metabolites (tetrahydroxyscopolamine, trihydroxy-methoxyscopolamine, and dihydroxy-dimethoxyscopolamine) are identified in rat urine. Seven metabolites (norscopine, scopine, tropic acid, aponorscopolamine, aposcopolamine, norscopolamine, and hydroxyscopolamine) and the parent drug are detected in rat plasma. Only 1 hydrolyzed metabolite (scopine) is found in the rat intestinal flora incubation mixture, and 2 metabolites (aposcopolamine and norscopolamine) are identified in the homogenized liver incubation mixture.