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OBJECTIVE To estab lish the pre paration method of clopidogrel active thiol metabolite （CATM），and to provide reference for the synthesis of cis-CATM. METHODS CATM was prepared ，separated and purified with isolated rat liver perfusion and ChromCore 120 C18 preparative column ，using（S）-2-oxo-clopidogrel as substrate. The target compounds were identified by mass spectrometry and nuclear magnetic resonance spectroscopy. The retention time of the active configuration of CATM in the human body （cis-CATM）were compared to confirm the proportion of active configuration in the target product. RESULTS The conversion rate of the target product was 11.71％. The target products were identified as CATM by MS and 1H-NMR. Peak 2-peak 5 of CATM were four stereoisomers. The retention time of them were 21.3，22.3，26.5，27.3 min. The peak area ratios of them were 7.13％，7.23％，63.52％，14.97％，respectively. Based on that retention time of the active configuration of CATM in human body was 26.3 min，the active cis-stereoisomer in the target product CATM accounted for 63.52％. CONCLUSIONS This method is low-cost ，simple，and can prepare CATM with higher active configuration.
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Objective@#To examine the effects of bisphenol A (BPA), bisphenol S ( BPS ), bisphenol F ( BPF ) and bisphenol AF ( BPAF ) on the proliferation and oxidative stress of BRL 3A rat liver cells, and to preliminarily evaluate their mutagenicities.@*Methods@#In vitro cultured BRL 3A rat liver cells were treated with BPA, BPS, BPF and BPAF at concentrations of 0, 5, 10, 25, 50, 100, 150 and 200 μmol/L for 48 h, respectively. Then, the cell viability was determined using the CCK-8 assay, and the half maximal inhibitory concentration ( IC50 ) was calculated. The minimum inhibitory concentration for BRL 3A cell proliferation was screened, and the intracellular reactive oxygen species ( ROS ) was measured in BRL 3A cells using the 2',7'-dichlorodihydrofluorescein diacetate ( DCFH-DA ) assay. In addition, the effects of BPA, BPS, BPF and BPAF at concentrations of 1 000, 200, 40, 8 and 1.6 μg/plate on the mutant colonies of histidine-deficient Salmonella typhimurium ( TA1535, TA97a, TA98, TA100 and TA102 ) were tested using the Ames test.@*Results@#Treatment with BPA and BPF at concentrations of 100 to 200 μmol/L and with BPAF at concentrations of 25 to 200 μmol/L inhibited BRL 3A cell survival at a concentration-dependent manner, while exposure to BPS at concentrations of 5 to 200 μmol/L resulted in no changes in BRL 3A cell survival. The IC50 values of BPA, BPS, BPF and BPAF were 131.7, >200, 187.5 and 21.6 μmol/L against BRL 3A cells, respectively. Treatment with BPS at 100 μmol/L or BPAF at 25 μmol/L caused no significant changes in the ROS level; however, exposure to BPA at 100 μmol/L and BPF at 100 μmol/L significantly increased the ROS level. Ames test showed that BPA, BPS, BPF and BPAF did not induce mutagenicity in TA1535, TA97a, TA98, TA100 or TA102 strains.@*Conclusions@#BPAF shows the highest cytotoxicity to BRL 3A cells, and low-concentration exposure to BPS has few effects on BRL 3A cells. The cytotoxicity of bisphenols against BRL 3A cells may be associated with the induction of oxidative stress. None of the four bisphenols show mutagenic effects under the present experimental conditions.
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Abstract Evaluating the effects of ecstasy on CYP2E1 activity is of great concern, mainly due to growing trends in abuse and co-administration of MDMA with ethanol and the dominant role of this isoenzyme on ethanol metabolism. This study aimed to evaluate the effects of MDMA on CYP2E1 activity. A total of 24 male rats were selected and divided into three groups. The first and second groups consisted of 12 rats and were employed to optimize the perfusion method, and the third group was employed for studying the alteration of CYP2E1 activity after liver exposure to MDMA (300 and 600 ng/ml). The amount of chlorzoxazone and 6-hydroxy chlorzoxazone in a sample obtained from liver perfusion before and after exposure to a buffer containing MDMA was determined by HPLC-FL. The enzymatic activity of rat CYP2E1 decreased after liver perfusion with a buffer containing 600 ng/ml of MDMA. However, no significant changes were observed in chlorzoxazone and 6-hydroxy chlorzoxazone concentration in perfusate before and after liver perfusion with a buffer containing 300 ng/ml of MDMA. Our findings suggest that the activity of CYP2E1 in rats might decrease only after administration of MDMA at a lethal dose. However, further animal and human studies are needed to confirm our assumption.
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OBJECTIVE：To i nvestigate the metabolism stabilities of novel hypoglycemic compound LSM- 13 in rat liver microsomes，and to analyze the possible metabolites. METHODS ：LSM-13 was dissolved in rat liver microsome incubation system initiated by reduced nicotinamide adenine dinucleotide phosphate ，and was incubated in water at 37 ℃. The reaction was terminated with acetonitrile at 0，5，10，15，30，45 and 60 min，respectively. Using indomethacin as internal standard ，the concentration of LSM-13 in incubation system was determined by HPLC. The residual percentage and enzyme kinetic parameters of LSM- 13 were calculated at different incubation time points with the concentration of LSM- 13 incubated for 0 min as reference. UPLC-Q-TOF/MS was used to analyze and speculate the metabolites of LSM- 13 in rat liver microsomes. RESULTS ：After 60 min incubation ，the remaining percentage of LSM- 13 was（56.07±0.95）％，the half-life was 42.78 min，and the intrinsic clearance was 0.032 4 mL/（min·mg）. Compared with total ion flow diagram of rat liver microsome blank samples ，three chromatographic peaks were added in the samples incubated for 60 min；the corresponding molecular ion peaks were m/z 505.133 8，417.102 4，293.111 7 [M+H]+；the possible metabolites may be dehydrogenation ，O-debentylation and hydrolysis products of LSM- 13. CONCLUSIONS ： The compound LSM- 13 has moderate stability in rat liver microsomes ，and may undergo dehydrogenation ，O-debentylation and hydrolysis.
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Objective To establish a rat liver transplantation model under direct vision of single operator and to explore the effect of different perfusion methods on the quality of the donor liver. Methods On the basis of the "two-cuff method" established by Kamada, the operation details were improved to established the rat liver transplantation model. The recipient rats were divided into two groups according to different perfusion methods, group A (perfusion via abdominal aorta) and group B (perfusion via portal vein). The perfusion effect, operation time, operation success rate, postoperative liver function, liver graft pathological manifestations and survival were compared between the two groups. Results There were more residual red blood cells in sinus hepaticus in group B than in group A after perfusion. Both the donor liver perfusion time and donor operation time were longer in group A than those in group B, and the differences were statistically significant (both P < 0.01). The success rate of operation in group A and group B was 77% and 71%, respectively. At 3 d after liver transplantation in rats, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TB) of the rats in the two groups were significantly higher than normal. At 7, 30 d after operation, compared with group A, the levels of ALT, AST and TB in group B were significantly increased, and the differences were statistically significant (all P < 0.01-0.05). The liver pathological examination showed that the degree of inflammatory reaction in the liver and degree of destruction of liver tissue in group B were more severe than those in group A, but there was no significant difference in long-term survival rate between the two groups. Conclusions Although the perfusion time and donor operation time of rat liver transplantation model were slightly prolonged by means of abdominal aorta perfusion, the perfusion effect was better, which can reduce liver tissue damage after operation and restore liver function to normal levels more quickly.
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To evaluate the anticancer potentials of Annona muricata fruit by in vitro and in vivo methods. Methods: The ethanolic extract of Annona muricata fruit was prepared by Soxhlet extraction method and further fractionated with petroleum ether, ethyl acetate and chloroform. The fractions were tested for cytotoxicity, apoptosis, scratch wound assay, and cell cycle analysis. IC50, apoptotic index and percentage cell migration were determined using HepG2 cells. For the in vivo studies, hepatocellular carcinoma was induced by administering 0.01% diethylnitrosamine (DEN) in drinking water in Wistar rats. In pre-treatment, rats were co-administered 200 mg/kg of fruit extract with DEN for 14 weeks. In post-treatment, the extract was co-administered after 8-weeks of DEN-induction for 14 weeks. Liver function test, haematological test, oxidative stress markers, relative liver weight, number of cancer nodules and histopathological parameters were determined. Results: Annona muricata fruit extract =significantly lowered cell proliferation counts. The chloroform-fraction possessed higher activity [IC50=(53.7±4.3) μg/mL]. The chloroform fraction inhibited cell migration, which was significant compared to curcumin. Further investigations regarding the mode of anticancer activity revealed that the chloroform fraction induced apoptosis. The cell cycle analysis indicated that cells were being arrested at G0/G1. In the in vivo studies, the DEN-control group showed a significant decrease in body weights with increased mortality rate, hepatic nodules, and impairment of liver function compared to normal rats. The rats pre-treated and post-treated with the extract showed positive results with significant improvement in the parameters that were adversely affected by DEN. In addition, other adverse effects of DEN, such as blood dyscrasias and hepatic endogenous antioxidant, were significantly attenuated by Annona muricata fruit extract. Conclusions: The Annona muricata fruit extract has anticancer activity when tested by in vitro and in vivo hepatocellular cancer models.
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To evaluate the anticancer potentials of Annona muricata fruit by in vitro and in vivo methods. Methods: The ethanolic extract of Annona muricata fruit was prepared by Soxhlet extraction method and further fractionated with petroleum ether, ethyl acetate and chloroform. The fractions were tested for cytotoxicity, apoptosis, scratch wound assay, and cell cycle analysis. IC50, apoptotic index and percentage cell migration were determined using HepG2 cells. For the in vivo studies, hepatocellular carcinoma was induced by administering 0.01% diethylnitrosamine (DEN) in drinking water in Wistar rats. In pre-treatment, rats were co-administered 200 mg/kg of fruit extract with DEN for 14 weeks. In post-treatment, the extract was co-administered after 8-weeks of DEN-induction for 14 weeks. Liver function test, haematological test, oxidative stress markers, relative liver weight, number of cancer nodules and histopathological parameters were determined. Results: Annona muricata fruit extract =significantly lowered cell proliferation counts. The chloroform-fraction possessed higher activity [IC50=(53.7±4.3) μg/mL]. The chloroform fraction inhibited cell migration, which was significant compared to curcumin. Further investigations regarding the mode of anticancer activity revealed that the chloroform fraction induced apoptosis. The cell cycle analysis indicated that cells were being arrested at G0/G1. In the in vivo studies, the DEN-control group showed a significant decrease in body weights with increased mortality rate, hepatic nodules, and impairment of liver function compared to normal rats. The rats pre-treated and post-treated with the extract showed positive results with significant improvement in the parameters that were adversely affected by DEN. In addition, other adverse effects of DEN, such as blood dyscrasias and hepatic endogenous antioxidant, were significantly attenuated by Annona muricata fruit extract. Conclusions: The Annona muricata fruit extract has anticancer activity when tested by in vitro and in vivo hepatocellular cancer models.
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@#A new method based on rat liver microsomes and chromatographic fingerprint comparison was established to investigate the possible herb-drug interactions between Liuwei Dihuang Pills(LDP)and nifedipine(NIF). LDP and NIF were incubated simultaneously with rat liver microsomes at 37 °C for 60 min in a shaking water bath. The separation was achieved on a C18 column using 0. 1% formic acid solution and acetonitrile as mobile phase with a liner gradient program. The flow rate was set at 1. 0 mL/min. Detection was achieved by UV light at 240 nm. To evaluate the interactions between LDP and NIF, the similarity of the fingerprinting chromatograms before and after co-incubation was calculated by similarity evaluation system for chromatographic fingerprint of TCM. Furthermore, pharmacokinetic experiments in rat suggested that there was no significant difference in the pharmacokinetic parameters of the typical components in LDP.
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OBJECTIVE: To investigate the hepatic toxicity of 8 monomers in Polygonum multiflorum using a combination of UDPglucuronic acid transferase 1A1(UGT1A1 enzyme). METHODS: Bilirubin was used as the substrate for UGT1A1. Incubation method in RLM in vitro was adopted to test the apparent inhibition constants(Ki) of different components. Furthermore the structure-activity relationship between the 8 components and UGT1A1 was analyzed. RESULTS: The inhibition effects on UGT1A1 enzyme of the 8 components were in the following sequence: emodin-8-O-glc > emodin > citreorosein > (+) -catechin > gallic acid > physcion > rhein > emodin-6-O-glc. Moreover, there was a structure-activity relationship, and it was presumed that the 6-position hydroxyl group is an active and necessary group. CONCLUSION: The established method in vitro is stable and feasible. Experimental results shows that the enzyme inhibition has structural selectivity, which provides an experimental basis for predicting the enzyme inhibition activity of the analogues of components of Polygonum multiflorum.
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Las recomendaciones de consumo de fibra no se cumplen y hay una necesidad por alimentos con fibra. El salvado de arroz (SA) tiene fibra y propiedades antioxidantes. Aquí se evaluaron estas propiedades en ratas suficientes (+) y deficientes (-) en Vitamina E (VitE) con o sin SA. Las ratas fueron divididas en cuatro grupos. Dos consumieron dietas +VitE y uno tenía SA. Los restantes consumieron dietas -VitE y uno tenía SA. El consumo de alimento, su eficiencia y el crecimiento, fueron similares entre los 4 grupos pero la masa fecal húmeda o seca fue 3 veces superior en los SA+. La hemoglobina en sangre y el hierro hepático fueron similares entre grupos, pero en los grupos (SA-) la VitE hepática fue 10 veces menor en las ratas -VitE que en las +VitE. Sin embargo, en las ratas -VitE/SA+, la VitE hepática fue sólo 2,6 veces menor. Este efecto del SA también se detectó en los eritrocitos, ya que la catalasa y la glutatión reductasa aumentaron en el grupo -VitE/SA-pero no en el grupo -VitE/SA+. El estudio muestra que SA no interfirió con el crecimiento y el metabolismo del hierro, sino que tuvo un efecto laxante y previno parcialmente la deficiencia de VitE.
Dietary fiber requirements are met by only a small fraction of the population. There is need for supplemented foods to fill this gap. Rice bran (RB) is high in fiber and has antioxidant properties. The effects of rice bran fiber on several metabolic indicators and the antioxidant capacity of rice bran in rats was reported. Rats were divided into 4 dietary groups: Vitamin E-sufficient with (+VitE/RB+) or without (+VitE/RB-) rice bran; Vitamin E-deficient with (-VitE/RB+) or without (-VitE/RB-) rice bran. Food intake, growth and feed efficiency were similar in all groups but wet and dry fecal mass of the RB+ groups were 3 times higher than the RB- groups. Blood hemoglobin and liver iron were also similar among all groups. However, the liver VitE concentration of the rats of (-VitE/RB-) group was 10x lower than the (+VitE/RB-) group. In contrast, liver VitE of the rats (-VitE/RB+) was only 2.6x lower. This effect of RB was also seen in erythrocytes since, catalase and glutathione reductase increased in the VitE deficient rats but RB prevented this increase. This study shows that dietary RB did not interfere with growth, feed efficiency and iron metabolism, it provided dietary fiber and laxation and partially prevented VitE deficiency.
As recomendações de ingestão de fibras não são cumpridos e existe uma necessidade de alimentos ricos em fibras. Farelo de arroz (FA) tem fibra e propriedades antioxidantes. Aqui, estas propriedades foram avaliadas em ratos suficientes (+) e pobres (-) em Vitamina E (VitE) com ou sem FA. Os ratos foram divididos em 4 grupos. Dois consumiram dietas +VitE e um tinha FA. Os restantes consumiram dietas -ViteE e um tinha FA. O consumo de alimento, sua eficiência e crescimento foram semelhantes entre os 4 grupos, mas nos grupos (FA-) a VitE hepática foi 10 vezes menor nos ratos -VitE que nos +VitE. Entretanto, nos ratos -VitE/FA+, a VitE hepática foi apenas 2,6 vezes menor. Este efeito do FA também foi detectado nos eritrócitos, visto que catalase e glutationa redutase aumentaram no grupo -VitE/FA-, mas não no grupo -VitE/FA+. O estudo mostra que FA não interferiu no crescimento ou no metabolismo do ferro, porém teve um efeito laxante e impediu parcialmente a deficiência de VitE.
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Animals , Mice , Oryza/adverse effects , Vitamin E Deficiency/etiology , Vitamin E/analysis , Antioxidants/analysis , Dietary Fiber , Eating , RatsABSTRACT
AIM To study the interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes.METHODS With maackiain and ephedrine as internal standards,HPLC was adopted in the simultaneous content determination of total tlavonoids (baicalin,wogonoside,baicalein,wogonin and oroxylin A) from Scutellariae Radix and total alkaloids (oxymatrine,oxysophocarpine and matrine)from Sophorae flavescentis Radix in whole prescription component (group) compatibility and single medicinal material part (group) at seven time points (0,15,30,45,60,90 and 120 min),followed by the calculation of their in vitro metabolic rates.RESULTS In whole prescription component (group) compatibility,the contents of baicalin and wogonoside were first increased (0-0.5 h) and then decreased (0.5-2.0 h),those of baicalein,wogonin and oroxylin A showed increasing trends (more obvious within 0.5 h).The stable and gende metabolisms of various alkaloids reached balance within 20 min.CONCLUSION The bioavailability improvement and efficacy enhancement of total flavonoids from Scutellariae Radix may attribute to the compatibility with total alkaloids from Sophorae flavescentis Radix and total polysaccharides from Rehmanniae Radix in Sanwu Huangqin Decoction
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AIM To study the interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes.METHODS With maackiain and ephedrine as internal standards,HPLC was adopted in the simultaneous content determination of total tlavonoids (baicalin,wogonoside,baicalein,wogonin and oroxylin A) from Scutellariae Radix and total alkaloids (oxymatrine,oxysophocarpine and matrine)from Sophorae flavescentis Radix in whole prescription component (group) compatibility and single medicinal material part (group) at seven time points (0,15,30,45,60,90 and 120 min),followed by the calculation of their in vitro metabolic rates.RESULTS In whole prescription component (group) compatibility,the contents of baicalin and wogonoside were first increased (0-0.5 h) and then decreased (0.5-2.0 h),those of baicalein,wogonin and oroxylin A showed increasing trends (more obvious within 0.5 h).The stable and gende metabolisms of various alkaloids reached balance within 20 min.CONCLUSION The bioavailability improvement and efficacy enhancement of total flavonoids from Scutellariae Radix may attribute to the compatibility with total alkaloids from Sophorae flavescentis Radix and total polysaccharides from Rehmanniae Radix in Sanwu Huangqin Decoction
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Background: This study investigates the knowledge, attitudes and practices of cosmetic surgery among female medical students at King Saud University (KSU). Methods: A quantitative observational cross-sectional approach was used to carry out the study at the KSU College of Medicine. A web-based questionnaire was first developed to collect the data necessary to fulfill the objectives of the research. The population under study included a random sample in which the questionnaire was sent to all female medical students at KSU. The sample size was estimated by using a single proportion formula with an acceptable margin of error at 5%. The sample size obtained was 384. Results: A response rate of 99% was achieved. The mean age of the participants was 20.9±1.48.Out of 381 KSU female medical students in our study, almost all participants (360, 94.5%) have heard about cosmetic surgery. Television was the source of knowledge for more than one third (38%) of participants who had already heard about cosmetic surgery. Just over half (51.4%) of surveyed KSU female medical students recognized the best definition of cosmetic surgery as “a surgery that modifies or improves the appearance of a physical feature electively”. A great majority of participants (86%) reported knowing of a particular type of cosmetic surgery, namely breast augmentation. Almost all participants (92.4%) agreed that “women perform more cosmetic surgery than men”. Only 9% of participants reported undergoing cosmetic surgery, where most of them (19 out of 35) went through laser treatment for the skin and almost the same number had a cosmetic surgery for personal satisfaction. Conclusion: From this research, we recommend further studies to go beyond educational institutions to the public at large, and to study different sub-populations.
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This mini-review summarises the risk factors for acquiring Respiratory Syncitial Virus (RSV) infection, and describes the harmful effects of the infection in pre-term infants. Moreover, theoretical considerations are discussed for the prevention of RSV infection in high-risk infant categories, such as pre-term infants. Background: Neonates positive for RSV are more prone to severe infection than neonates infected with other common respiratory viruses. Despite RSV infection being more common in late neonates than in early ones, pre-term infants ≤ 35 wk gestational age (GA) are at high risk for developing severe RSV disease. Efforts to prevent infection include case management, vaccination and the identification of risk factors. The morbidity and mortality risks of RSV disease are highest in pre-term newborns with other underlying disease, such as bronchopulmonary dysplasia (BPD) or hemodynamically significant congenital heart disease (hsCHD). Associations between RSV-positive neonates and climate factors are also discussed. Nosocomial-acquired respiratory syncytial virus infections in pre-term infants in Neonatal Intensive Care Units (NICUs) are reported. The development of an RSV vaccine has been challenging, and vaccine in pre-term infants is currently unavailable. Palivizumab, a monoclonal antibody licensed for the prevention of RSV, lowers respiratory tract disease in pre-term infants. The home healthcare nurse can play an important role. By developing patient and caregiver trust, the nurse can implement an RSV prevention plan, leading to a decrease in the hospitalization of premature infants with RSV. Conclusions: Commercially-insured late pre-term infants with RSV infection are at high risk of recurrent wheezing and infantile asthma for 1 year after the initial episode, and pose a significant economic burden on the healthcare system. Education is critical for the continuing development of clinical NICU nursing practice.
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This study aimed to investigate the effects of tannins extracted from Tibetan medicine Phyllanthi Fructus on cytochrome P450 enzyme of liver microsomes in rats.Cocktail probe substrates were incubated with liver microsomes in vitro.Metabolic elimination percentages of the six probe substrates,including dapsone,dextromethorphan hydrobromide,coumarin,phenacetin,chlorzoxazone and tolbutamide,were determined by HPLC.The effects of tannins extracted from Tibetan medicine Phyllanthi Fructus on the enzymatic activity of rat liver microsomal P450s was evaluated.It was found that tannins extracted from Phyllanthi Fructus did not impact P450 enzymes.The IC50 values of CYP3A4 and CYP1A2 were over 200 μg·mL-1,while the IC50 value of CYP2C9 was superior to 500 tg·mL-1.In conclusion,Tannins extracted from Tibetan medicine Phyllanthi Fructus did not significantly affect cytochrome P450 enzymes.
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OBJECTIVE: To identify the specific cytochrome P450 (CYP) enzymes involved in the metabolism of dipfluzine hydrochloride (Dip) in the rat liver microsomes. METHODS: The rat liver microsomes were prepared and incubated with Dip. The Dip metabolites (M1, M2, M4 and M5) were identified by LC-MS/MS, and the CYP isoenzymes were identified by the combination of the selective CYP inhibitor study, correlation analysis and a panel of recombinant rat CYP expereiment, respectively. RESULTS: The results from the experiments of selective CYP inhibitors, correlation analysis and recombinant rat CYP isoenzymes indicated that CYP2A1, CYP3A and CYP2C11 contributed to the formation of M1 and M5 in the rat liver microsomes. CYP3A, CYP2A1, CYP1A2, CYP2C11 and CYP2E1 metabolized Dip to M2. CYP3A, CYP2A1, CYP2E1 and CYP2C11 contributed to M4 formation. And the recombinant rat CYP researches further indicated that CYP3A2 exhibited more activity than CYP3A1. CONCLUSION: CYP3A and CYP2A1 are the major CYP isoenzymes responsible for catalyzing Dip to the four metabolites formation in the rat liver microsomes.
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OBJECTIVE: To evaluate the effects of dipfluzine hydrochloride (Dip) on CYP450s activities in vitro and in vivo in rats. METHODS: Markers were incubated in the normal rat liver microsomes with Dip (0-200 μmol·L-1) and the concentration of metabolites of the markers were determined by LC-MS/MS, and then the ratios were calculated to evaluate the effects of Dip on the CYP450s activities. Dip was administered by orally to the male SD rats at doses of 30, 60 and 90 mg·kg-1 body weight and phenobarbital was administered at doses of 120 mg·kg-1 body weight for 14 d. At the fifteenth day, the rats were orally administered the Cocktail probe markers and blood samples were collected via medial angle of eye at different time. The concentrations of markers were determined by LC-MS/MS and the drug-time curve was plotted, by which the pharmacokinetic parameters were calculated. And then the rat liver microsomes were prepared and the probe markers were added into the incubation samples. The concentrations of the probe markers and its metabolites were determined and the metabolism ratios were calculated. The effects of Dip on CYP450s activities were evaluated by comparing the following outcomes between the experimental groups and the control group: the relative liver weight, the concentrations of protein, the contents of CYP450 enzymes, the drug concentration-time curves, the pharmacokinetic parameters and the metabolism ratios of the probes. RESULTS: In the normal rat liver microsomes, Dip had the inhibitive effects on CYP2D1, CYP2C6 and CYP2C11, and the IC50 were 8.85, 20.93 and 69.45 μg·mL-1, respectively. Dip had no effect on the relative weight of livers, the protein concentrations and the CYP450 content for the rats after they were fed on Dip for 14 d, but these indexes were raised remarkably when phenobarbital was administered by orally to rats. The results displayed that the low-dose Dip inhibited the activity of CYP2D1 or induced the activity of CYP2C11 in rats. Moderate-dose Dip showed the abilities to inhibit CYP2D1, but induce CYP2C11 and CYP3A. High-dose Dip had the certain inhibitive effects to CYP2C6 and CYP2D1, and had the inductive effects on CYP2C11 and CYP3A. CYP1A2, CYP2C11, CYP2C12, CYP2D1 and CYP3A were all induced after administration of phenobarbital. CONCLU SION: The results of liver microsomes incubation and blood plasma from the normal and Dip-induced rats all show that Dip inhibit the activities of CYP2C6 and CYP2D1, however, Dip inhibit CYP2C11 in vitro and induce it in vivo.
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Objective Re Du Ning Injection (RDN), a Chinese materia medica injection, is made from the extracts of Lonicerae Japonicae Flos, Gardeniae Fructus, and Artemisiae Annuae Herba. Since last decade, RDN has been widely used in China for the treatment of viral infection, fever, and inflammation. To assess the potential interacting of RDN with co-administered drugs, the inhibitory effects of RDN on the enzyme activities (CYP1A1, CYP1A2, CYP2C11, CYP2D1, and CYP3A1/2) of rat liver microsomes were investigated by a cocktail method. Methods A sensitive and specific LC-MS method capable of simultaneous quantification of five metabolites in rat liver microsomes was developed and validated. Then RDN (0.625%–1.0%) was incubated with rat liver microsomes and specific substrates. The enzyme activities were expressed as the formation rate of the specific metabolites of the substrates (pmol · mg · protein−1 · min−1). Results RDN competitively inhibited the activities of CYP1A2 and CYP2C11, with inhibition constant (Ki) values determined to be 0.18% and 0.63%, respectively. RDN exhibited the mixed inhibition on the activity of CYP2D1, with a Ki value of 0.15%. The activities of CYP1A1 and CYP3A1/2 were not markedly inhibited even by 1.0% RDN. Conclusion RDN could inhibit the rat enzyme activities of CYP1A2, 2C11, and 2D1 in vitro with different inhibition modes, which is worthy of promoting safety and efficacy of RDN.
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Aim: Our case report aims to inform practicing clinicians of an unusual presentation of vitamin C deficiency in the setting of a developed nation where this illness is rare and underappreciated. Case Presentation: We present the case of a noncompliant 16-year-old African American female with vertically transmitted human immunodeficiency virus/acquired immune deficiency syndrome who presented to the emergency department (ED) with a CD4 count of 4 and a hemoglobin level of 5.7 g/dL. In the ED, she was found to have persistent low-grade bleeding initially believed to be of an upper gastrointestinal origin, but which was later found come from the oral mucosa. Her stools were dark in color and guaiac positive. She was hemodynamically unstable, for which she was transfused with packed red blood cells and briefly treated with continuous norepinephrine infusion. Her initial coagulation studies were noncontributory with an international normalized ratio of 1.1, a prothrombin time of 35, and a platelet count that was also within normal limits. Esophagogastroduodenoscopy and a colonoscopy were both unremarkable. Bone marrow biopsy showed normocellular marrow with 80% cellularity and trilinear hematopoiesis. Her vitamin C level was zero. She was diagnosed with scurvy and treated with vitamin C supplementation. Discussion and Conclusion: Vitamin C deficiency can lead to an often-forgotten medical condition called scurvy. It can cause defective collagen synthesis leading to fragile capillaries, gingival bleeding, and cutaneous changes. Unrecognized, this condition can lead to significant bleeding and can be lethal in select patient populations. Our case is unique in that it shows that vitamin C deficiency can masquerade as upper gastrointestinal bleeding and may present with significant hemodynamic instability requiring blood transfusions and vasopressor support. It is therefore imperative to keep in mind the diagnosis of scurvy as a potential cause of hemodynamic instability even in an industrialized nation such as the United States. Vitamin C deficiency is a rare and underdiagnosed medical entity in the hospital setting that can lead to hemodynamic instability. Scurvy patients can present with melena and oral bleeding, mimicking hematemesis.
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This study was aimed to investigate the induction effect ofRe-Du-Ning (RDN) Injection on rat liver microsome CYP450 enzymes. SD rats were randomly divided into the solvent control group, positive control group as well as the low, middle and high dose group of RDN (1, 2, 4 mL·kg-1·d-1). After drugs were administrated continuously for 7 days, the rats were sacrificed. The liver was weighed and prepared to microsomes. Meanwhile, the liver coefficients of rats were calculated. And the protein content was detected by BCA method. Finally, activities of five important subtypes of CYP450 enzymes such as CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A1/2 were measured by the“cocktail” method. The results showed that the levels of liver coefficients, microsome yield rate and activities of CYP450 subtypes increased significantly in the positive control group compared with the solvent control group (P < 0.01). There was no significant difference on the levels of liver coefficients, microsome yield and protein content between the low and middle dose group of RDN. However, there was significant difference on the levels of liver coefficients and microsome yield in the high dose group (P < 0.05). In terms of the influence on enzyme activity, RDN Injection can significantly induce the activities of CYP1A2 with dose dependence. It can induce the activities of CYP2C9 and CYP2C19 at the middle and high dose. However, there was no obvious influence on the activities of CYP3A1/2 and CYP2D6. It was concluded that the positive control group can obviously induce activities of CYP450, which can be used in the evaluation of induction experiments. RDN Injection had induction effect on CYP1A2, CYP2C9 and CYP2C19. But it had no influence on the activities of CYP3A1/2 and CYP2D6.