基于网络药理学和体内实验探讨茯苓多糖抗对乙酰氨基酚致肝损伤的作用机制

目的利用网络药理学和体内实验,探讨茯苓多糖(PCP)抗对乙酰氨基酚(APAP)致肝损伤的作用靶点。方法通过SwissTargetPrediction数据库、SuperPred数据库和Pharmmapper数据库获取PCP的作用靶点,利用OMIM数据库、GeneCards数据库查找APAP致肝损伤相关作用靶点,使用String数据库构建蛋白互作网络(PPI)并筛选核心靶点。利用R软件进行基因本体(GO)和京都基因与基因组百科全书(KEGG)通路分析,通过AutoDuck vina软件对茯苓多糖和核心靶点进行分子对接。最后,用HE染色评估小鼠肝组织损伤情况,并且通过RT-PCR和免疫组化染色法(IHC)检测核心靶点的表达情况。结果共获得26个核心靶点,GO富集分析显示细胞凋亡、细胞周期、免疫等生物过程可能参与PCP抗APAP致肝损伤的进程。KEGG富集分析显示茯苓多糖抗APAP致肝损伤的信号通路主要集中在p53信号通路(p53 signaling pathway)、NOD样受体信号通路(NOD-like receptor signaling pathway)、TNF信号通路(TNF signaling pathway)等。分子对接结果显示,PCP与等核心靶点具有较好的结合能力。HE染色显示,APAP组小鼠肝组织形态结构出现异常,表现为细胞大面积坏死,而PCP干预后可以减轻肝组织损伤。RT-PCR和IHC结果表明,PCP通过逆转APAP暴露引起的CDK2、TNF、BCL2L1及MAOA水平升高来缓解小鼠肝损伤。结论通过网络药理学和体内实验找到了PCP抗APAP致肝损伤的可能潜在靶点,其作用机制可能与调节细胞周期、免疫反应、凋亡和药物代谢有关。

紫外标准曲线法测定对乙酰氨基酚片的不确定度评定

采用紫外标准曲线法测定对乙酰氨基酚片含量,评估测定结果的不确定度。通过不确定度来源分析,建立数学模型,量化不确定度分量,计算出测定值的不确定度结果及各分量的贡献率。不确定度的主要来源为系列标准溶液的配制及样品处理过程,其次为紫外仪器和标准工作溶液的配制。当k=2时,按置信区间为95%,对乙酰氨基酚片百分标示量含量测定值为(99.55±2.48)%。不确定度评估可以用于分析和优化紫外标准曲线法测定药品含量的过程,对药品的质量控制和方法改进有重要的指导意义。

对乙酰氨基酚-腺苷蛋氨酸复方制剂对大鼠慢性肝损伤的作用

目的探讨对乙酰氨基酚(APAP)-腺苷蛋氨酸(SAMe)复方制剂对大鼠慢性肝损伤的作用。方法60只SD雄性大鼠随机均分为正常对照组(生理盐水)、APAP组(210 mg/kg APAP)、低剂量APAP+SAMe组(210 mg/kg APAP+53 mg/kg SAMe)、中剂量APAP+SAMe组(210 mg/kg APAP+105 mg/kg SAMe)、高剂量APAP+SAMe组(210 mg/kg APAP+210 mg/kg SAMe)及阳性对照组[210 mg/kg APAP+200 mg/kg N-乙酰半胱氨酸(NAC)],各组给予相应药物灌胃1次/d,连续30 d,观察给药期间各组大鼠一般情况(精神状态、行为活动、饮食情况、二便情况及皮肤毛色等);末次给药16 h后,10%水合氯醛麻醉各组大鼠,腹主动脉取血,采用多功能酶标仪检测各组大鼠血清中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆红素(TBiL)及乳酸脱氢酶(LDH)的含量;取血后于冰面上取各组大鼠肝脏、称重并计算肝脏指数,采用苏木素-伊红(HE)染色法制备切片观察各组大鼠肝组织的组织学特征,制备10%肝组织匀浆检测谷胱甘肽(GSH)、超氧化物歧化酶(SOD)及脂质过氧化产物丙二醛(MDA)含量。结果与正常对照组相比,APAP组大鼠血清ALT、AST、TBiL及LDH水平明显上升(P<0.01),肝组织中MDA水平明显升高、SOD和GSH活性明显降低(P<0.01),肝组织切片存在明显的损伤,肝脏指数明显升高(P<0.01);APAP+SAMe不同剂量组大鼠肝损伤有不同程度的改善(P<0.05或P<0.01)。结论APAP配伍SAMe使用能明显降低长期使用APAP带来的肝损伤。

Photoelectrochemical degradation of acetaminophen and valacyclovir using nanoporous titanium dioxide

Electrochemically treated nanoporous TiO2 was employed as a novel electrode to assist in the pho- toelectrochemical degradation of acetaminophen and valacyclovir. The prepared electrode was characterized by scanning electron microscopy （SEM） and energy dispersive spectroscopy （EDS）. Cyclic voltammetry （CV）, Mott-Schottky plots, ultraviolet-visible light （UV-vis） absorbance spec- troscopy, and a total organic carbon （TOC） analyzer were employed to investigate the photoelec- trochemical degradation of acetaminophen and valacyclovir. The results indicated no obvious re- moval of acetaminophen and valacyclovir over 3 h when separate photochemical degradation and electrochemical oxidation were employed. In contrast, acetaminophen and valacyclovir were rapid- ly eliminated via photoelectrochemical degradation. In addition, electrochemically treated nanopo~ rous TiO2 electrodes significantly enhanced the efficacy of the photoelectrochemical degradation of acetaminophen and valacyclovir, by 86.96% and 53.12%, respectively, when compared with un- treated nanoporous TiO2 electrodes. This enhanced performance may have been attributed to the formation of Ti3~, Ti2~, and oxygen vacancies, as well as an improvement in conductivity during the electrochemical reduction process. The effect of temperature was further investigated, where the activation energy of the photoelectrochemical degradation of acetaminophen and valacyclovir was determined to be 9.62 and 18.42 kJ/mol, respectively.

Plasma microRNA profiles distinguish lethal injury in acetaminophen toxicity: A research study

AIM: To investigate plasma microRNA (miRNA) profiles indicative of hepatotoxicity in the setting of lethal acetaminophen (APAP) toxicity in mice. METHODS: Using plasma from APAP poisoned mice, either lethally (500 mg/kg) or sublethally (150 mg/kg) dosed, we screened commercially available murine microRNA libraries (SABiosciences, Qiagen Sciences, MD) to evaluate for unique miRNA profiles between these two dosing parameters. RESULTS: We distinguished numerous, unique plasma miRNAs both up- and downregulated in lethally compared to sublethally dosed mice. Of note, many of the greatest up- and downregulated miRNAs, namely 574-5p, 466g, 466f-3p, 375, 29c, and 148a, have been shown to be associated with asthma in prior studies. Interestingly, a relationship between APAP and asthma has been previously well described in the literature, with an as yet unknown mechanism of pathology. There was a statistically significant increase in alanine aminotransferase levels in the lethal compared to sublethal APAP dosing groups at the 12 h time point (P < 0.001). There was 90% mortality in the lethally compared to sublethally dosed mice at the 48 h time point (P = 0.011). CONCLUSION: We identified unique plasma miRNAs both up- and downregulated in APAP poisoning which are correlated to asthma development.

Effect of Lianshu preparation on lipopolysaccharide-induced diarrhea in rats

AIM： To investigate the effect of Lianshu preparation on lipopolysaccharide （LPS）-induced diarrhea in rats. METHODS： A diarrhea model was established in Sprague Dawley rats via injection of 1 mL of 30 mg/kg LPS. A total of 40 rats were randomly divided into normal group, LPS group, LPS ＋ Lianshu group, LPS ＋ berberine group （n = 10 in each group）. Their intestinal mucosal barrier and frequency of diarrhea were observed. Levels of glucose, serum Na^＋, K^＋, Cl and hematocrit, plasma nitrogen monoxide （NO）, diamine oxidase （DAO）, and D （-）-lactate were measured. The number of IgA＋ plasma cells in small intestine was detected and SIgA levels in the intestinal fluid were measured. The antipyretic activity of Lianshu preparation in rats was evaluated using Brewer＇s yeast-induced pyrexia （10 mL/kg of 20% aqueous suspension）. Acetaminophen （250 mg/kg, intragastric administration, bid） was comparison. Temperature used as a standard drug for was recorded 1 h before and 6 h after Brewer＇s yeast injection. Finally, small intestina transmission in mice treated with Lianshu was detected after intraperitoneal injection of methyl prostigmin （2 mg/kg）. Atropine （10 g/kg） was used as a control. The ink content in intestine was determined and the total length of intestine was measured. RESULTS： The frequency of diarrhea was higher in LPS group than in LPS ＋ Lianshu group and LPS ＋ berberine group （36.70± 5.23 vs 28.50 ±4.06 and 32.70±9.30 respectively, P 〈 0.01）, and lower in LP5 ＋ Lianshu group than in LPS ＋ berberine group （P = 0.03）. The levels of Na＋, glucose, Cl, K^＋ were significantly lower in LPS ＋ Lianshu group than in LPS ＋ berberine group （140.35±3.19 mmol/L vs 131.99±4.86 mmol/L, 8.49 ±1.84 mmol/L vs 6.54±2.30 mmol/L, 106.29± 4.41 mmol/L vs 102.5±1.39 mmol/L, 5.08±0.66 mmol/L vs 4.32 ± 0.62 mmol/L respectively, P 〈 0.05）. The level of hematocrit was lower in LPS ＋ Lianshu group than in LPS ＋ berberine group （0.50% ±0.07% vs 0.59%± 0.10% respectively, P 〈 0.05）. The plasma levels of NO, DAO and D （-）-lactate were higher in LPS group than in normal group （79.74 ± 7.39μmol/L vs 24.94 ± 3.38μmol/L, 2.48 ±0.42μ/mL vs 0.82 ±0.33 p/mL, 5.63± 0.85μg/mL vs 2.01 ±0.32 μg/mL respectively, P 〈 0.01）, and lower in LPS ＋ Lianshu group than in LP5 ＋ berberine group （48.59±4.70μmol/L vs 51.56 ±8.38 μmol/L, 1.43± 0.53μmol/mL vs 1.81 ±0.42 μmol/mL, 4.00± 0.54 μg/mL vs 4.88 ± 0.77 pg/mL respectively, P 〈 0.05）. The morphology of the intestinal mucosa showed destroyed villi in LPS group and atrophied intestinal mucosa in other groups. The pathological intestinal mucosal changes were less in LPS ＋ Lianshu group than in LPS group. The number of IgA＋ plasma cells and amount of SIgA were higher in LPS ＋ Lianshu group than in LPS group （1.16±0.19/μm^2 vs 1.09±0.28/μm^2, P = 0.026; 0.59 ±0.12 mg/L vs 0.15± 0.19 mg/L respectively, P = 0.000）. Lianshu had counteractive effects on yeast-induced pyrexia and enterokinesia in rats. CONCLUSION： Lianshu preparation has therapeutic effects on LPS-induced diarrhea and enterokinesia in rats.

HPLC法测定感冒灵胶囊中各组分的含量

采用高效液相色谱法同时测定感冒灵胶囊中各组分含量。方法简便、快速、准确。

Dynamic tracking of stem cells in an acute liver failure model

AIM： To investigate a dual labeling technique, which would enable real-time monitoring of transplanted em- bryonic stem cell （ESC） kinetics, as well as long-term tracking. METHODS： Liver damage was induced in C57/BL6 male mice （n = 40） by acetaminophen （APAP） 300 mg/kg administered intraperitoneally. Green fluores- cence protein （GFP） positive C57/BL6 mouse ESCs were stained with the near-infrared fluorescent lipophilic tracer 1,1-dioctadecyl-3,3,3,3-tetramethylindotricarbo- cyanine iodide （DiR） immediately before transplantationinto the spleen. Each of the animals in the cell therapy group （n = 20） received 5 x 106 ESCs 4 h following treatment with APAP. The control group （n = 20） re- ceived the vehicle only. The distribution and dynamics of the cells were monitored in real-time with the IVIS lumina-2 at 30 rain post transplantation, then at 3, 12, 24, 48 and 72 h, and after one and 2 wk. Immunohisto- chemical examination of liver tissue was used to identify expression of GFP and albumin. Plasma alanine amino- transferase （ALT） was measured as an indication of liver damage.RESULTS： DiR-stained ESCs were easily tracked with the IVIS using the indocyanine green filter due to its high background passband with minimal background autofluorescence. The transplanted cells were confined inside the spleen at 30 min post-transplantation, gradu- ally moved into the splenic vein, and were detectable in parts of the liver at the 3 h time-point. Within 24 h of transplantation, homing of almost 90% of cells was confirmed in the liver. On day three, however, the DiR signal started to fade out, and ex vivo IVIS imaging of different organs allowed signal detection at time-points when the signal could not be detected by in vivo imag- ing, and confirmed that the highest photon emission was in the liver （P 〈 0.0001）. At 2 wk, the DiRsignal was no longer detectable in vivo; however, immuno- histochemistry analysis of constitutively-expressed GFP was used to provide an insight into the distribution of the cells. GFP ＋ve cells were detected in tissue sections resembling hepatocytes and were dispersed throughout the hepatic parenchyma, with the presence of a larger number of GFP ＋ve cells incorporated within the sinu- soidal endothelial lining. Very faint albumin expression was detected in the transplanted GFP ＋re cells at 72 h; however at 2 wk, few cells that were positive for GFP were also strongly positive for albumin. There was a significant improvement in serum levels of ALT, albumin and bilirubin in both groups at 2 wk when compared with the 72 h time-point. In the cell therapy group, serum ALT was significantly （P = 0.016） lower and al- bumin （P = 0.009） was significantly higher when com- pared with the control group at the 2 wk time-point;however there was no difference in mortality between the two groups. CONCLUSION： Dual labeling is an easy to use and cheap method for longitudinal monitoring of distribu- tion, survival and engraftment of transplanted cells, and could be used for cell therapy models.

Protective effect of tea polyphenols against paracetamol-induced hepatotoxicity in mice is significanly correlated with cytochrome P450 suppression

AIM： To investigate the hepatoprotective activity of tea polyphenols （TP） and its relation with cytochrome P450 （CYP450） expression in mice. METHODS： Hepatic CYP450 and CYPbs levels were measured by UV-spectrophotometry in mice 2 d after intraperitoneal TP （25, 50 and 100 mg/kg per day）. Then the mice were intragastricly pre-treated with TP （100, 200 and 400 mg/kg per day） for six days before paracetamol （1000 mg/kg） was given. Their acute mortality was compared with that of control mice. The mice were pre-treated with TP （100, 200, and 400 mg/kg per day） for five days before paracetamol （500 mg/kg） was given. Hepatic CYP2E1 and CYPIA2 protein and mRNA expression levels were evaluated by Western blotting, immunohistochemical staining and transcriptase-polymerase chain reaction. RESULTS： The hepatic CYP450 and CYPb5 levels in mice of TP-treated groups （100, 200 and 400 mg/kg per day） were decreased in a dose-dependent manner compared with those in the negative control mice.TP significantly attenuated the paracetamol-induced hepatic injury and dramatically reduced the mortality of paracetamol-treated mice. Furthermore, TP reduced CYP2E1 and CYPIA2 expression at both protein and mRNA levels in a dose-dependent manner. CONCLUSION： TP possess potential hepatoprotective properties and can suppress CYP450 expression.

Factors associated with irritable bowel syndrome symptoms in hemodialysis patients

AIM: To investigate clinical characteristics associated with the presence of irritable bowel syndrome (IBS) symptoms in hemodialysis (HD) patients.METHODS: This was a cross-sectional study. A questionnaire based on the Bowel Disease Questionnaire that records gastrointestinal symptoms was given to 294 patients in 4 dialysis centers. A total of 196 (67%) subjects returned the survey. A multivariable logistic regression model was used to identify factors significantly associated with IBS symptoms. RESULTS: Symptoms compatible with IBS were present in 27 (13.8%) subjects and independently associated with low post-dialysis serum potassium [OR = 0.258, 95% CI (0.075-0.891), P = 0.032], paracetamol use [OR = 3.159, 95% CI (1.214-8.220), P = 0.018], and Kidney Disease Quality of Life (KDQOL) cognitive function score [OR = 0.977, 95% CI (0.956-0.999), P = 0.042]. Univariate regressions were also performed and the reported significance is for multivariate analysis. No association was detected for age, gender, depressed mood, smoking (present or past), body mass index, albumin level, Kt/V, sodium preor post-dialysis level, change in potassium level during HD, proton pump inhibitor or H2 blocker use, aspirin use, residual diuresis, hepatitis B or C infection, diabetes mellitus, marital status and education level. CONCLUSION: This study examined potential risk factors for symptoms compatible with IBS in HD patients and identified an association with paracetamol use, post-dialysis potassium level and KDQOL-cognitive function score.

Protective effects of 2,4-dihydroxybenzophenone against acetaminophen-induced hepatotoxicity in mice

AIM:To examine the effects of 2,4-dihydroxybenzophenone(BP-1),a benzophenone derivative used as an ultraviolet light absorbent,on acetaminophen(APAP)induced hepatotoxicity in C57BL/6J mice.METHODS:Mice were administered orally with BP-1 at doses of 200,400 and 800 mg/kg body weight respectively every morning for 4 d before a hepatotoxic dose of APAP(350 mg/kg body weight) was given subcutaneously.Twenty four hours after APAP intoxication,the serum enzyme including serum alaine aminotransferase(ALT),aspartate aminotransferase(AST),lactate dehydrogenase(LDH) were measured and liver histopathologic changes were examined.RESULTS:BP-1 administration dramatically reduced serum ALT,AST and LDH levels.Liver histopathological examination showed that BP-1 administration antagonized APAP-induced liver pathological damage in a dose-dependent manner.Further tests showed that APAP-induced hepatic lipid peroxidation was reduced significantly by BP-1 pretreatment,and glutathione depletion was ameliorated obviously.CONCLUSION:BP-1 can effectively protect C57BL/6J mice from APAP-induced hepatotoxicity,and reduction of oxidative stress might be part of the protection mechanism.

Protective effects of 5-methoxypsoralen against acetaminophen-induced hepatotoxicity in mice

AIM:To investigate the hepatic protective effects of 5-methoxypsoralen(5-MOP) and to learn if 5-MOP causes hepatotoxicity at protective doses.METHODS:C57BL/6J mice were administrated orally with 5-MOP at doses of 12.5,25 and 50 mg/kg body weight respectively every morning for 4 d before given acetaminophen(APAP) subcutaneously at a dose of 500 mg/kg.The 5-MOP alone group was treated with 5-MOP orally at a dose of 50 mg/kg body weight for 4 d without APAP.Twenty-four hours after APAP administration,blood samples of mice were analyzed for serum enzyme alanine transaminase(ALT),aspartate transaminase(AST),lactate dehydrogenase(LDH) levels,and malondialdehyde(MDA),reduced glutathione(GSH) and oxidized glutathione(GSSG) of liver tissues were measured and histopathologic changes of the liver were observed.RESULTS:Compared with the vehicle control group,the serum levels(IU/L) of ALT,AST and LDH were all increased significantly in APAP group(8355 ± 3940 vs 30 ± 21,P < 0.05;6482 ± 4018 vs 146 ± 58,P <0.05;24627 ± 10975 vs 1504 ± 410,P < 0.05).Compared with APAP group,the serum ALT levels(IU/L)(1674 ± 1810 vs 8355 ± 3940,P < 0.05;54 ± 39 vs 8355 ± 3940,P < 0.05;19 ± 9 vs 8355 ± 3940,P < 0.05),AST levels(IU/L)(729 ± 685 vs 6482 ± 4108,P < 0.05;187 ± 149 vs 6482 ± 4108,P < 0.05;141 ± 12 vs 6482 ± 4108,P < 0.05) and LDH levels(IU/L)(7220 ± 6317 vs 24 627 ± 10 975,P < 0.05;1618 ± 719 vs 24 627 ± 10 975,P < 0.05;1394 ± 469 vs 24 627 ± 10 975,P < 0.05) were all decreased drastically in the three-dosage 5-MOP pretreatment groups.Pretreatment of 5-MOP could attenuate histopathologic changes induced by APAP,including hepatocellular necrosis and infiltration of inflammatory cells,and the effect was dose-dependent.MDA levels(nmol/mg) were decreased by 5-MOP in a dose-dependent manner(0.98 ± 0.45 vs 2.15 ± 1.07,P > 0.05;0.59 ± 0.07 vs 2.15 ± 1.07,P < 0.05;0.47 ± 0.06 vs 2.15 ± 1.07,P < 0.05).The pretreatment of 5-MOP could also increase the GSH/GSSG ratio(3.834 ± 0.340 vs 3.306 ± 0.282,P > 0.05;5.330 ± 0.421 vs 3.306 ± 0.282,P < 0.05;6.180 ± 0.212 vs 3.306 ± 0.282,P < 0.05).In the group treated with 5-MOP but without APAP,the serum enzyme levels,the liver histopathologic manifestation,and the values of MDA and GSH/GSSG ratio were all normal.CONCLUSION:5-MOP can effectively protect C57BL/6J mice from APAP-induced hepatotoxicity and possesses an antioxidative activity,and does not cause liver injury at the protective doses.

Impaired gluconeogenesis in a porcine model of paracetamol induced acute liver failure

AIM:To investigate glucose homeostasis and in particular gluconeogenesis in a large animal model of acute liver failure(ALF).METHODS:Six pigs with paracetamol induced ALF under general anaesthesia were studied over 25 h.Plasma samples were withdrawn every five hours from a central vein.Three animals were used as controls and were maintained under anaesthesia only.Using 1 H NMR spectroscopy we identified most gluconeogenic amino acids along with lactate and pyruvate in the animal plasma samples.RESULTS:No significant changes were observed in the concentrations of the amino acids studied in the animals maintained under anaesthesia only.If we look at the ALF animals,we observed a statistically significant rise of lactate(P<0.003)and pyruvate(P<0.018) at the end of the experiments.We also observed statistically significant rises in the concentrations of alanine(P<0.002),glycine(P<0.005),threonine(P< 0.048),tyrosine(P<0.000),phenylalanine(P<0.000) and isoleucine(P<0.01).Valine levels decreased significantly(P<0.05).CONCLUSION:Our pig model of ALF is characterized by an altered gluconeogenetic capacity,an impaired tricarboxylic acid(TCA)cycle and a glycolytic state.

The effects of acetaminophen combined with radiation on the radiosensitivity of irradiated human glioma cell progeny

Objective： To study the effects of acetaminophen （ACE） combined with radiation on the progeny of the human glioma cell line SHG-44, and to investigate if ACE may be an useful therapeutic radiosensitivity agent in the treatment of recurrent human glioma. Methods： A randomized, controlled experiment, was performed at the Department of Radiology Laboratory, the First Hospital Affiliated to Soochow University, between September 2004 and January 2006. Brain glioma SHG-44 cells were divided into three groups： SHG-44, SHG-44-10, and SHG-44-10 ＋ ACE cells groups. The SHG-44-10 cells group was irradiated with dose of 10 Gy by a linear accelerator （6 MVX）. It was passaged for 15 generations and cultured in RPMI-1640 culture media. Then SHG-44-10 ＋ ACE cells group was treated with ACE. Measures： Community re-double time, mean lethal dose （DO）, extrapolation number （N）, fraction surviving fraction irradiated by 2 Gy dose （SF2）, quasi-threshold dose （Dq）, and cell cycle. Results： The SF2 of the SHG-44, SHG-44-10, and SHG-44-10 ＋ ACE cells groups were 70.8%, 80.6% and 45.2%, respectively, with significance （P = 0.040）. The SHG-44-10 and SHG-44-10 ＋ ACE cells groups were irradiated with 8 Gy. After 12 hours, the G2/M ratio of the SHG-44-10 and SHG-44-10 ＋ ACE cells groups were indicating significantly higher ratio compared to pre-irradiated groups （P 〈 0.01）. After 24 hours, the G2/M ratio of the SHG-44-10 cells group decreased rapidly, while the ratio of the SHG-44-10 ＋ ACE cells group still maintained in high level. Conclusion： In the present study, Subtoxic dose of ACE increased the radiosensitivity of the progeny of irradiated human glioma cell. ACE may be an useful radiosensitivity agent in the treatment of recrudescent human malignant glioma.

The Hepatoprotective Effects of Solanum Incanum on Acetaminophen-lnduced Hepatotoxicity in Guinea Pigs

Solanum incanum, a shrubby herb, is widely distributed and used as analgesic, antitoxic, and antispasmodic in folk medicine. In the present study, the protective effects of aqueous extract of S. incanum against acetaminophen induced acute liver damage were evaluated in guinea pigs. Animals were orally administered with S. incanum extract （50 and 100 mg/ kg bw） and silymarin （ 100 mg / kg bw） respectively for 6 days followed by acetaminophen administration （2 g / kg bw） at the 7th day. The results showed that the treatment with S. incanum extract significantly lowered the acetaminophen-induced serum levels of hepatic marker enzymes （AST, ALT, and ALP）, Liver histopathology also showed that S. incanum extract reduced the incidence of liver lesions including the swelling of hepatic cells, lymphocytes infiltration, nucleus condensation, and hepatic necrosis induced by acetaminophen treatment in guinea pigs. The S. incanum extract at a dose of 100 mg / kg bw was more effective in suppressing the oxidative damage than the extract at a dose of 50 mg / kg bw. Therefore, the results of this study suggest that S. incanum extract could protect liver against the acetaminophen-induced oxidative damage.

Antioxidative and antiapoptotic effects of （＋）-clausenamide on acetaminophen-induced nephrotoxicity in mice

Objective： （＋）-Clausenamide （（＋）-CLA）, the active ingredient of wampee, was isolated from the leaves of Clausena lansium （Lour.） Skeels. This study aimed to evaluate the protective potential of （＋）-CLA against acetaminophen （APAP）-induced nephrotoxicity in mice. Methods： Mice were divided into control, APAP, high-dose （＋）-CLA, and low-dose （＋）-CLA groups. Then, mice were preadministered （＋）-CLA （50 and 100 mg/kg） for 5 consecutive days. After the last treatment, the animals received a single intraperitoneal injection of APAP （600 mg/kg）. Renal histopathology was evaluated by staining with hematoxylin and eosin. The levels of malondialdehyde （MDA） and glutathione （GSH） and the activities of catalase （CAT） and superoxide dismutase （SOD） were determined using corresponding kits. Western blotting was used to analyze the expression of apoptosis-related proteins in renal tissue. Results： Administration of APAP increased serum creatinine and blood urea nitrogen levels in comparison with the control group. An increase in renal MDA level, depletion of GSH, and reductions in CAT and SOD activities in renal tissue indicated that APAP-induced kidney injury was mediated by oxidative stress. The expressions of Bax and caspase-3, cleavage of caspase-3, and cytoplasm cytochrome c levels were up-regulated in renal tissue, whereas Bcl-2 expression and mitochondrial cytochrome c levels were down-regulated in the APAP group, which revealed that APAP-induced kidney injury significantly increased cell apoptosis in renal tubules. The histopathology of kidney tissue supported these biochemical mechanisms. （＋）-CLA can reverse changes in most of the abovementioned parameters and nearly restore the normal structure of the kidney. Conclusion： Oxidative stress and apoptosis are considered to be the mechanisms underlying APAP-induced nephrotoxicity. （＋）-CLA could be a promising antidote for APAP-induced acute renal damage owing to its antioxidative and antiapoptotic effects.

Crystal Structure of 2-(4-Fluorophenylamino)-2-thiomethyl acetophenone

The title compound 2-(4-fluorophenylamino)-2-thiomethyl acetophe-none (C15H14FNOS, M. = 275. 34 ) was synthesized by the reaction of 2-hydroxy-2-thiomethyl acetohenone (abenzoylhemithioacetal ) with 4-fluoroaniline. The crystal ismonoclinic, space group P21/n. with unit cell constants a = 12. 434 (4 ), b = 6. 00l2(6), c=l8. 49l(2) A, β=100. 87(l)° Z = 4, V = 1355. O(5) A, D'= 1. 350g/cm3, μ(MoKa) = 2. 41 cm-l, F(000) = 576, R=0. 029, Rw.= 0. 041 for 14l2 ob-served reflections (I> 3σ(I) ). X-ray analysis reveals that the dihedral angle betweenplane l (C(l)~C(6), F, N ) and plane 2 (C(8)~C(14), O ) is 4. l8° and theC(7) -S bond is longer than normal single C-S bond.

Chlorogenic acid prevents acetaminophen-induced liver injury: the involvement of CYP450 metabolic enzymes and some antioxidant signals

Chlorogenic acid（CGA）, a polyphenolic compound, is abundant in fruits, dietary vegetables, and some medicinal herbs. This study investigated the prevention of CGA against acetaminophen（AP）-induced hepatotoxicity and its engaged mechanisms. CGA reversed the decreased cell viability induced by AP in L-02 cells in vitro. In addition, CGA reduced the AP-induced increased serum levels of alanine/aspartate aminotransferase（ALT/AST） in vivo. The effect of CGA on cytochrome P450（CYP） enzymatic（CYP2E1, CYP1A2, and CYP3A4） activities showed that CGA caused very little inhibition on CYP2E1 and CYP1A2 enzymatic activities, but not CYP3A4. The measurement of liver malondialdehyde（MDA）, reactive oxygen species（ROS）, and glutathione（GSH） levels showed that CGA prevented AP-induced liver oxidative stress injury. Further, CGA increased the AP-induced decreased m RNA expression of peroxiredoxin（Prx） 1, 2, 3, 5, 6, epoxide hydrolase（Ephx） 2, and polymerase（RNA） II（DNA directed） polypeptide K（Polr2k）, and nuclear factor erythroid-2-related factor 2（Nrf2）. In summary, CGA ameliorates the AP-induced liver injury probably by slightly inhibiting CYP2E1 and CYP1A2 enzymatic properties. In addition, cellular important antioxidant signals such as Prx1, 2, 3, 5, 6, Ephx2, Polr2 k, and Nrf2 also contributed to the protection of CGA against AP-induced oxidative stress injury.

Acetaminophen-induced hepatotoxicity in rats is correlated with the accumulation of bile acids: an underlying mechanism

In the present study, we aimed to investigate the underlying mechanism of acetaminophen（APAP）-induced hepatotoxicity by measuring the expression levels of liver transporters and concentrations of bile acids（BAs） in rat plasma and liver. SD rats（42） were randomly assigned into six groups, including 6-h control group, APAP 6-h group, 12-h control group, APAP 12-h group, 24-h control group and APAP 24-h group. The estimation study of BAs in plasma and liver was performed on LC-MS/MS. The levels of bile salt export pump（Bsep）, multidrug resistant protein 2（Mrp2）, multidrug resistant protein 4（Mrp4）, Na＋/taurocholate cotransporting polypeptide（Ntcp） and organic anion transporting polypeptide 2（Oatp2） in the liver were analyzed by Western blotting analysis. Compared with the corresponding control groups, no difference was found in the BA levels and the expressions of BA transporters in the plasma and liver after 6 h of APAP administration. While BA levels were significantly decreased in the plasma and increased in the liver after 12 h of APAP administration（P0.05）; and the expressions of Bsep and Mrp2 were significantly reduced（P0.05）. After 24 h of APAP administration, BA levels were both greatly increased in the plasma and liver（P0.05）; and the expressions of Mrp4 and Oatp2 were significantly decreased（P0.05）. In response to over-dose APAP, Bsep, Mrp2, Mrp4 and Oatp2 levels were reduced at different time points, causing the accumulation of BAs, and such accumulation may ultimately lead to the severe liver injury, which could be an underlying mechanism of the APAP-induced hepatotoxicity.

Analysis of alternation with ibuprofen and acetaminophen in outpatient and emergency departments of a children’s hospital

In the present study,we retrospectively analyzed 105382 prescriptions of antipyretic drugs for children(0–14 years of age)in the outpatient and emergency departments of Ningbo Women and Children’s Hospital from January to December 2019.Ibuprofen and acetaminophen are relatively safe and the most commonly used drugs in daily life.Due to different dosage forms,there are acetaminophen suspension drops,acetaminophen oral solution,ibuprofen suspension,and ibuprofen suppositories for children.Through investigation,we found that the age of antipyretic drugs used by children in our hospital ranged from 3 months to 14 years old,which could be divided into two groups:single drug use and alternating use of ibuprofen and acetaminophen,with an alternating use rate of 3.19%.There were differences between the two groups in terms of age,gender,the proportion of emergency departments use,and the amount of single prescription(P<0.05).The age of the alternating use group was younger,and the male proportion,the proportion of emergency departments use,single prescription amount of the alternating use group were higher.In the alternating use group,ibuprofen and acetaminophen of different dosage forms were used alternatively.After pair comparison analysis,they only had an age difference(P<0.008),which was in line with the principle that different dosage forms are suitable for different ages.The method of alternation with ibuprofen and acetaminophen is mainly used in the emergency departments and children of younger age because most of the children have a refractory fever,which is complicated and changes relatively fast.There was no record of adverse reactions about alternation of ibuprofen and acetaminophen in the software system of adverse reactions.However,this study was a retrospective analysis,and there were risks of missing records.Therefore,alternating use of ibuprofen and acetaminophen still couldn’t ignore the risk of adverse reactions.