OBJECTIVE: To investigate the hepatotoxicity of alkaloids from Euodiae Fructus combined with berberine (BBR) in Zuojin Pill, and to preliminarily explore the possible detoxification mechanism of the combination components. METHODS: The combination ratio of components was determined by the maximum concentration (Cmax) of the chemical components in Zuojin Pill. HepG2 cell model was used to investigate the combined toxicity of the hepatotoxic components from Euodiae Fructus, such as evodiamine (EVO) or dehydroevodiamine (DHED), with BBR for 48 h. The experimental groups were set as follows: the vehicle control group, the EVO group, the DHED group, the BBR group, and the combination group of EVO or DHED with BBR. The cell counting kit-8 (CCK-8) method was used to determine the cell viability, and the combination index (CI) was used to determine the combined toxicity of the components. The alanine transaminase (ALT), aspartate aminotransferase (AST), lactate dehydroge-nase (LDH), and alkaline phosphatase (ALP) activities as well as total bilirubin (TBIL) content in the cell culture supernatant were detected. The protein expression levels of bile acid transporters, such as bile salt export pump (BSEP) and multidrug resistance-associated protein 2 (MRP2), were detected by Western blot. The intracellular malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in HepG2 cells were detected. RESULTS: Compared with EVO or DHED group, the combination of EVO 1 μmol/L with BBR 10 μmol/L or DHED 50 μmol/L with BBR 35 μmol/L significantly increased cell viability of HepG2 cells (P < 0.01), with CI values of 77.89 or 4.49, respectively, much greater than 1. Significant decreases in the activities of ALT, AST, LDH, ALP, and TBIL content in the cell culture supernatant were found in both combination groups (P < 0.05, P < 0.01). Compared with the EVO group, the combination of EVO with BBR upregulated the protein expression levels of BSEP and MRP2. Compared with the DHED group, the combination of DHED with BBR significantly downregulated the protein expression levels of BSEP and MRP2 (P < 0.01). Compared with EVO or DHED group, the combination of EVO or DHED with BBR significantly reduced the MDA content in HepG2 cells (P < 0.05, P < 0.01). CONCLUSION A certain ratio of BBR combined with EVO or DHED had an antagonistic effect on HepG2 cytotoxicity, which might be related to regulating the expression of bile acid transpor-ters, and reducing lipid peroxidation damage.
Humans ; Hep G2 Cells ; Berberine/pharmacology* ; Drugs, Chinese Herbal/toxicity* ; Evodia/chemistry* ; Alkaloids/pharmacology* ; Cell Survival/drug effects* ; Multidrug Resistance-Associated Proteins/metabolism* ; Multidrug Resistance-Associated Protein 2 ; Quinazolines

Objective:To investigate whether escin(ESC)and dextromethorphan(DEX)have the protective effects on the progression and symptoms of Alzheimer disease(AD).Methods:The AD model of Caenorhabditis elegans(C.elegans)was established by transgenic amyloid β-protein(Aβ protein).Different concentrations of ESC or DEX or 50 μmol/L memantine(MEM)were used to treat the AD model worms,and their lifespan was detected.The movement ability of AD model C.elegans was evalua-ted by body bending frequency and head swinging frequency.The changes in cognitive functions of AD model C.elegans before and after treatment were detected by chemotaxis experiments.The changes in A βprotein and reactive oxygen species(ROS)content in C.elegans were detected.The changes in gene pathways related to oxidative stress were detected by Real-time quantitative polymerase chain reaction(RT-qPCR).Results:At high dose 1 000 μmol/L,ESC or DEX treatment showed no significant effects on the activity of C.elegans.Compared with untreated worms,the survival time of AD model C.elegans in the 20 μmol/L ESC and 60 μmol/L DEX intervention groups was significantly extended.In the middle stage of AD progression,the body bending frequency and head swinging frequency of AD model worms after ESC or DEX treatment was significantly increased compared with the untreated control group with DEX being more effective in the recovery of head swinging frequency.For the early cognitive function tests,the chemotaxis index of ESC or DEX treated worms was significantly higher than that of the untreated worms,which correlated with marked reductions in the Aβ protein levels.The reactive oxygen species content in the drug intervention group was also lower than that in the control group.RT-qPCR results showed that ESC could inhibit oxidative stress in the AD model C.elegans by a 2-fold upregulation of skn1 expression.Conclusion:ESC and DEX could improve the reductions of movement ability and cognitive function in the AD model worms and delay the aggravation of AD-related symptoms.ESC delays the progression of AD pos-sibly by activating the SKN-1/Nrf2 pathway to protect against oxidative injury in the AD model.

Objective To establish a gas chromatography-mass spectrometry method for simultaneous determination of the contents of volatile components muscone,menthol,borneol,isoborneol,camphor,and carvone in Wuli Huichun pills,and to evaluate the quality of Wuli Huichun pills based on authenticity,effectiveness and safety.Methods The ethyl acetate extract was analyzed by HP-5MS capillary column(30 m×0.25 mm,0.25 μm)with temperature programming.The injection port temperature was set at 230℃,and the split ratio was set at 10∶1.The carrier gas was high-purity helium gas.The injection port was in constant flow mode,with a carrier gas flow rate of 1.0 mL·min-1.The detector was a mass spectrometer with a triple quadrupole in series.The ion source was an electron bombardment source(EI)with a temperature of 250℃.The collision gas was argon.The temperature of the mass spectrometry transmission interface was 250℃.Scanning mode was SIM mode,with a solvent delay of 2 minutes.Results The linear relationships of the six components were good(r>0.999 0),the average recoveries were 93.57%-97.96%,and the RSD was 0.73%-2.5%.Conclusion The method is simple,accurate,and efficient,and can be used for the quality evaluation of Wuli Huichun pills.

Objective:To investigate whether escin(ESC)and dextromethorphan(DEX)have the protective effects on the progression and symptoms of Alzheimer disease(AD).Methods:The AD model of Caenorhabditis elegans(C.elegans)was established by transgenic amyloid β-protein(Aβ protein).Different concentrations of ESC or DEX or 50 μmol/L memantine(MEM)were used to treat the AD model worms,and their lifespan was detected.The movement ability of AD model C.elegans was evalua-ted by body bending frequency and head swinging frequency.The changes in cognitive functions of AD model C.elegans before and after treatment were detected by chemotaxis experiments.The changes in A βprotein and reactive oxygen species(ROS)content in C.elegans were detected.The changes in gene pathways related to oxidative stress were detected by Real-time quantitative polymerase chain reaction(RT-qPCR).Results:At high dose 1 000 μmol/L,ESC or DEX treatment showed no significant effects on the activity of C.elegans.Compared with untreated worms,the survival time of AD model C.elegans in the 20 μmol/L ESC and 60 μmol/L DEX intervention groups was significantly extended.In the middle stage of AD progression,the body bending frequency and head swinging frequency of AD model worms after ESC or DEX treatment was significantly increased compared with the untreated control group with DEX being more effective in the recovery of head swinging frequency.For the early cognitive function tests,the chemotaxis index of ESC or DEX treated worms was significantly higher than that of the untreated worms,which correlated with marked reductions in the Aβ protein levels.The reactive oxygen species content in the drug intervention group was also lower than that in the control group.RT-qPCR results showed that ESC could inhibit oxidative stress in the AD model C.elegans by a 2-fold upregulation of skn1 expression.Conclusion:ESC and DEX could improve the reductions of movement ability and cognitive function in the AD model worms and delay the aggravation of AD-related symptoms.ESC delays the progression of AD pos-sibly by activating the SKN-1/Nrf2 pathway to protect against oxidative injury in the AD model.

Objective To establish a gas chromatography-mass spectrometry method for simultaneous determination of the contents of volatile components muscone,menthol,borneol,isoborneol,camphor,and carvone in Wuli Huichun pills,and to evaluate the quality of Wuli Huichun pills based on authenticity,effectiveness and safety.Methods The ethyl acetate extract was analyzed by HP-5MS capillary column(30 m×0.25 mm,0.25 μm)with temperature programming.The injection port temperature was set at 230℃,and the split ratio was set at 10∶1.The carrier gas was high-purity helium gas.The injection port was in constant flow mode,with a carrier gas flow rate of 1.0 mL·min-1.The detector was a mass spectrometer with a triple quadrupole in series.The ion source was an electron bombardment source(EI)with a temperature of 250℃.The collision gas was argon.The temperature of the mass spectrometry transmission interface was 250℃.Scanning mode was SIM mode,with a solvent delay of 2 minutes.Results The linear relationships of the six components were good(r>0.999 0),the average recoveries were 93.57%-97.96%,and the RSD was 0.73%-2.5%.Conclusion The method is simple,accurate,and efficient,and can be used for the quality evaluation of Wuli Huichun pills.

OBJECTIVE To investigate the impact of mercuric sulfide nanoparticles(HgS-NP)on various neurotransmitter-type neurons in Caenorhabditis elegans(C.elegans).METHODS Wild-type N2 C.elegans,as well as the transgenic C.elegans expressing green fluorescent protein specifically in gamma-amino-butyric acid(GABA)-ergic,glutamatergic,dopaminergic,cholinergic,and serotoninergic neurons(strains EG1285,DA1240,BZ555,LX929,and GR1366),were synchronized at the L4 stage and treated with different concentrations of HgS-NP via solid exposure.① Five transgenic strains of C.elegans were exposed to HgS-NP 0(control group),250,500,and 1000 mg·L-1 for 72 h.Fluorescence microscopy was used to observe the expression of green fluorescence in different neurotransmitter-type neurons of the corresponding transgenic C.elegans.② Wild-type N2 C.elegans were exposed to HgS-NP 0(control group)and 1000 mg·L-1 for 72 h.Real-time quantitative PCR(RT-qPCR)was performed to detect the mRNA expression levels of 33 genes related to the GABAergic and glutamatergic neurotransmitter systems.RESULTS ① Compared with the control group of the same genotype,the EG1285 C.elegans exhibited soma shrinkage,dendrite fragmentation,and neuronal loss,with a significant decrease in the relative fluorescence intensity of GABAergic neurons(P<0.05)after exposure to different concentra-tions of HgS-NP.After exposure to HgS-NP 1000 mg·L-1,the DA1240 C.elegans showed a loss of glutamatergic neurons in the head region and a significant decrease in the relative fluorescence intensity(P<0.01).However,there were no significant changes in the morphology or relative fluorescence intensity of dopaminergic,cholinergic,and serotoninergic neurons in BZ555,LX929,and GR1366 C.elegans after exposure to HgS-NP,respectively.② Compared with the control group,wild-type N2 C.elegans exposed to HgS-NP 1000 mg·L-1 showed increased mRNA expression levels of glr-7 and glr-8,which encoded non-N-methyl-D-aspartate glutamate receptors(P<0.05),while the expression levels of the remaining 31 genes related to the GABAergic and glutamatergic neurotransmitter systems showed no significant changes.CONCLUSION Exposure to a high dose of HgS-NP for 72 h may potentially damage the GABAergic and glutamatergic neurons in C.elegans,but have no significant effect on dopaminergic,cholinergic or serotoninergic neurons.

Objective:In order to understand the changing trends of gastric cancer incidence and mortality in early-onset and late-onset in China from 2000 to 2019.Methods:The Global Burden of Disease research data was collected, and Excel and R 4.2.1 softwares were used to examine the incidence rate, mortality rate, and disability-adjusted life years (DALY) of Chinese people from 2000 to 2019, with a focus on gender, age, and year.Results:In 2019, the crude incidence rates were 7.06/100 000 (95% UI: 6.63/100 000-7.59/100 000) and 114.52/100 000 (95% UI: 108.79/100 000-121.63/100 000) for early- and late-onset gastric cancer, respectively. The crude mortality rate for early-onset gastric cancer was 3.29/100 000 (95% UI: 3.11/100 000- 3.50/100 000), while the crude mortality rate for late-onset gastric cancer was 81.88/100 000 (95% UI: 78.15/100 000-86.04/100 000). Additionally, the crude DALY rates for these two types of gastric cancer were 156.48/100 000 (95% UI: 148.82/100 000-165.84/100 000) and 1 750.13/100 000 (95% UI: 1 661.21/100 000-1 852.99/100 000). The standardized incidence of early-onset gastric cancer decreased from 5.49/100 000 in 2000 to 4.76/100 000 in 2019, and that of late-onset gastric cancer decreased from 143.45/100 000 in 2000 to 123.02/100 000 in 2019.The standardized mortality rate of early-onset gastric cancer decreased from 4.16/100 000 in 2000 to 2.18/100 000 in 2019, and that of late-onset gastric cancer decreased from 140.82/100 000 in 2000 to 91.49/100 000 in 2019. The standardized DALY rate for early-onset gastric cancer in 2019 was 105.87/100 000 (95% UI: 87.98/100 000 -125.60/100 000), lower than 198.84/100 000 (95% UI: 179.47/100 000- 219.83/100 000) in 2000. The standardized DALY rate for late onset gastric cancer in 2019 was 1 821.11/100 000 (95% UI: 1 509.42/100 000-2 158.53/100 000), lower than 2 932.52/100 000 (95% UI: 2 665.92/100 000-3 252.60/100 000) in 2000. Conclusions:The standardized mortality rate of early-onset gastric cancer in China showed a decreasing trend from 2000 to 2019. The standardized mortality rate of late onset gastric cancer showed a trend of first increasing and then decreasing. Notably, the incidence, mortality, and DALY of late-onset gastric cancer were significantly higher than those of early-onset gastric cancer during this period. Additionally, male incidence, mortality, and crude DALY rates were higher than female.

Objective To investigate clinical effects of hyperbaric oxygen (HBO) combined with sensitive antibiotic alveolar wash on severe craniocerebral injury complicated with pulmonary infection.Methods One hundred and eighty-seven patients with severe craniocerebral injury who were admitted to the Blue Cross Brain Hospital for treatment from January 2017 to November 2018 were selected as research subjects.The diagnostic criteria of all the recruited patients were in accordance with the expert consensus on patients with severe infection of China Neurological Surgery.The patients were randomized into 3 groups:the HBO + diluted antibiotic alveolar wash group (or group Ⅰ) (63 cases),the diluted antibiotic alveolar wash group (or group Ⅱ) (63 cases) and venous antibiotic infusion group (or group Ⅲ) (61 cases).After 30 days of treatment with the above-mentioned treatment profiles,arterial blood gas partial pressure,changes in GCS scores,mechanical ventilation time,length of ICU stay and clinical effects both before and after treatment were closely observed and compared between the 3 groups.Results After 3 courses of treatment,the levels of arterial blood gas partial pressure and GCS scores were all obviously increased as compared with those before treatment,and statistical significance could be seen when comparisons were made between them (P ＜ 0.01).The arterial blood gas partial pressure of group Ⅰ was significantly higher than that of the 2 other groups,also with statistical significance (P ＜ 0.05).The mechanical ventilation time and length of ICU stay of the patients of group Ⅰ were considerably shorter than those of group Ⅱ and group Ⅲ,also with statistical significance (P ＜ 0.05 or P ＜ 0.01).Total efficacy rate of group Ⅰ (92.1％) was obviously higher than that those of group Ⅱ (84.1％)and group Ⅲ (77.1),also with statistical significance (P ＜ 0.05).Conclusion HBO combined with sensitive antibiotic alveolar wash in treatment of severe craniocerebral injury complicated with pulmonary infection could achieve significant effects.For this reason,it is worth further clinical extension.

Objective To investigate clinical effects of hyperbaric oxygen (HBO) combined with sensitive antibiotic alveolar wash on severe craniocerebral injury complicated with pulmonary infection.Methods One hundred and eighty-seven patients with severe craniocerebral injury who were admitted to the Blue Cross Brain Hospital for treatment from January 2017 to November 2018 were selected as research subjects.The diagnostic criteria of all the recruited patients were in accordance with the expert consensus on patients with severe infection of China Neurological Surgery.The patients were randomized into 3 groups:the HBO + diluted antibiotic alveolar wash group (or group Ⅰ) (63 cases),the diluted antibiotic alveolar wash group (or group Ⅱ) (63 cases) and venous antibiotic infusion group (or group Ⅲ) (61 cases).After 30 days of treatment with the above-mentioned treatment profiles,arterial blood gas partial pressure,changes in GCS scores,mechanical ventilation time,length of ICU stay and clinical effects both before and after treatment were closely observed and compared between the 3 groups.Results After 3 courses of treatment,the levels of arterial blood gas partial pressure and GCS scores were all obviously increased as compared with those before treatment,and statistical significance could be seen when comparisons were made between them (P ＜ 0.01).The arterial blood gas partial pressure of group Ⅰ was significantly higher than that of the 2 other groups,also with statistical significance (P ＜ 0.05).The mechanical ventilation time and length of ICU stay of the patients of group Ⅰ were considerably shorter than those of group Ⅱ and group Ⅲ,also with statistical significance (P ＜ 0.05 or P ＜ 0.01).Total efficacy rate of group Ⅰ (92.1％) was obviously higher than that those of group Ⅱ (84.1％)and group Ⅲ (77.1),also with statistical significance (P ＜ 0.05).Conclusion HBO combined with sensitive antibiotic alveolar wash in treatment of severe craniocerebral injury complicated with pulmonary infection could achieve significant effects.For this reason,it is worth further clinical extension.

OBJECTIVETo study characteristics of energy metabolism in the skeletal muscle of rats with postoperative fatigue syndrome (POFS) and the interventional effect of ginsenoside Rb1.

METHODWe chose resection of 70% of the "middle" small intestine as the rat model for POFS. Ninety-six adult male SPF SD rats were randomly divided into the control group, the model group, and the ginsenoside Rb1-treated group by body weight. And then, each group was further randomly divided into four subgroups, according to different postoperative investigated time points, such as postoperative day 1, postoperative day 3, postoperative day 7 and postoperative day 10. So the animals were divided into twelve subgroups (n = 8 in each subgroup). Rats of the control group and the model group were injected intraperitoneally with saline at the dose of 10 mL x kg(-1) one hour before the operation and once a day during the postoperative days. Rats of the ginsenoside Rb1-treated group were administered 10 mg x kg(-1) ginsenoside Rb1 by the same method. The skeletal muscles were sampled on postoperative day 1, 3, 7 and 10. The contents of ATP, ADP, AMP in skeletal muscles were determined by HPLC, and the activities of Na(+)-K(+)-ATPase and Ca(2+)-ATPase were investigated by colorimetry.

RESULTCompared with the control group, the content of ATP in skeletal muscle of rats of the model group decreased significantly on postoperative day 3 (P < 0.05), while the content of ADP significantly increased on postoperative day 7 and 10 (P < 0.05). The activity of Na(+)-K(+)-AT-Pase decreased on postoperative day 3 and 7 (P < 0.05), and the activity of Ca(2+)-ATPase decreased on postoperative day 7. After supplement of ginsenoside Rb1, on the investigated time points, all the negative changes of the indicators discovered above were significantly adjusted (P < 0.05) in rats of the ginsenoside Rb1-treated group, while no significant differences were investigated.

CONCLUSIONDuring a certain period of postoperative time, the activity of energy metabolism is depressed in the skeletal muscle of rats with POFS, but it can be improved by supplement of ginsenoside Rb1.

Animals ; Calcium-Transporting ATPases ; physiology ; Energy Metabolism ; drug effects ; Fatigue ; drug therapy ; metabolism ; Ginsenosides ; pharmacology ; therapeutic use ; Male ; Muscle, Skeletal ; metabolism ; Postoperative Complications ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; physiology ; Syndrome