Mesaconitine was incubated with rat liver microsomes in vitro. The metabolites of mesaconitine in rat liver microsomes were identified by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method with high resolution power. A typical reaction mixture of 100 mol L-1 Tris-HCI buffer (pH 7.4) containing 0.5 gL-1 microsomal protein and 50 micro molL-1 mesaconitine was prepared. The above reaction mixture was divided into six groups, and the volume of each group was 200 micro L. The incubation mixture was pre-incubated at 37 degrees C for 2 min and the reactions were initiated by adding NADPH generating system. After 90 min incubation at 37 degrees C, 200 micro L of acetonitrile was added to each group to stop the reaction. The metabolites of mesaconitine were investigated by UPLC-MS/MS method. Mesaconitine and 6 metabolites M1-M6 were found in the incubation system. The structures were characterized according to the data from MS/MS spectra and literatures. The metabolic reactions of mesaconitine in rat liver microsomes included the demethylation, deacetylation, dehydrogenation and hydroxylation. The major metabolic pathways of mesaconitine in rat liver microsomes were determined by UPLC-MS/MS on multiple reaction monitoring (MRM) mode combined with specific inhibitors of cytochrome P450 (CYP) isoforms, including alpha-naphthoflavone (CYP1A2), quinine (CYP2D), diethyldithiocarbamate (CYP2E1), ketoconazole (CYP3A) and sulfaphenazole (CYP2C), separately. Mesaconitine was mainly metabolized by CYP3A. CYP2C and CYP2D were also more important CYP isoforms for the metabolism reactions of mesaconitine, but CYP1A2 and CYP2E1 haven't any contribution to MA metabolism in rat liver microsomes.
Aconitine ; analogs & derivatives ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 CYP3A Inhibitors ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Enzyme Inhibitors ; pharmacology ; Ketoconazole ; pharmacology ; Male ; Metabolic Networks and Pathways ; Microsomes, Liver ; enzymology ; metabolism ; Quinine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfaphenazole ; pharmacology ; Tandem Mass Spectrometry

Using a UPLC-MS/MS (MRM) and cocktail probe substrates method, the metabolic fingerprint of the compatibility of Radix Aconite (RA) and Radix Paeoniae Alba (RPA) and its effect on CYP450 enzymes were investigated. These main CYP isoforms include CYP 1A2, CYP 2C, CYP 2E1, CYP 2D and CYP 3A. Compared with the inhibition effect of RA decoctions on CYP450 isoforms, their co-decoctions of RA and RPA with different proportions can decrease RA' inhibition on CYP3A, CYP2D, CYP2C and CYP1A2, but can not reduce RA' effect on CYP2E1. The metabolic fingerprints of RA decoction and co-decoctions with different proportions of RPA in CYP450 of rat liver were analyzed by UPLC-MS. Compared with the metabolic fingerprints of RA decoction, the intensity of diester-diterpenoid aconitum alkaloids decreased significantly, while the intensity of monoester-diterpenoid alkaloids significantly increased in the metabolic fingerprints of co-decoctions of RA and RPA. The results suggest that RA coadministration with RPA increased the degradation of toxic alkaloid and show the effect of toxicity reducing and efficacy enhancing.
Aconitum ; chemistry ; Alkaloids ; chemistry ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 Enzyme Inhibitors ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Liver ; drug effects ; enzymology ; Metabolome ; Paeonia ; chemistry ; Rats ; Tandem Mass Spectrometry

Objective:To analyze the clinical characteristics that affect the early outcomes of the elderly patients with transient ischemic attack(TIA) and provide the guidance for the prediction of disease recurrence.Methods:From January 2016 to December 2019, 315 TIA patients over 60 years old based on the time-clinical definition who were admitted to the department of neurology in Beijing Shijitan Hospital were divided into recurrence cerebrovascular events group(RCVEs group) and non-recurrence cerebrovascular events group(non-RCVEs group) according to the recurrence at 1 month after onset. The clinical characteristics of the two groups were compared by Logsitic regression analysis.Results:There were 29 cases in the RCVEs group and 286 cases in the non-RCVEs group. Compared to the non-RCVEs group, patients in RCVEs group were more likely have higher prevalence of hypertension history(14/15, 83/203), moderate or severe intracranial artery stenosis(21/8, 132/154), carotid stenosis(22/7, 107/179), DWI positive, motor symptoms, carotid artery stenting and higher times of TIA episodes(2(2, 3), 2(1, 2)), fasting blood glucose and glycosylated hemoglobin levels(8.86(8.17, 9.56)mmol/L, 5.82(5.27, 6.33)mmol/L). Logistic regression analysis showed that patients in RCVEs group had a DWI positive( B=1.634, OR=5.124, 95% CI: 2.898-9.059, P=0.000) and higher prevalence of moderate to severe stenosis of carotid artery( B=0.572, OR=1.773, 95% CI: 1.050-2.993, P=0.032). Conclusion:DWI positive and moderate to severe carotid stenosis may influence the early poor prognosis in patients with TIA.

Objective: To observe the clinical efficacy of moxibustion therapy plus Liu's pediatric massage (tuina) for children with recurrent respiratory tract infections due to qi deficiency of spleen and lung. Methods: A total of 60 children who met the inclusion criteria were divided into an observation group and a control group according to the visiting sequence, with 30 cases in each group. Children in the observation group were treated with moxibustion therapy plus Liu's pediatric massage, and those in the control group were treated with Liu's pediatric massage alone. The incidence of respiratory tract infections and traditional Chinese medicine (TCM) symptoms score were observed and recorded in both groups before and after treatment. And the clinical efficacy was compared between the two groups. Results: The total effective rate of the observation group was 93.3％, and that of the control group was 83.3％. The difference between the two groups was statistically significant (P<0.05). After treatment, the TCM symptoms score and total times of infections in both groups were all statistically different from those before treatment (all P<0.05). The differences in TCM symptoms score and infection frequency before and after treatment in the observation group were statistically different from those in the control group (both P<0.05). Conclusion: Moxibustion therapy plus Liu's pediatric massage has a better effect in improving the clinical symptoms and reducing the frequency of respiratory tract infections for children with recurrent respiratory tract infections due to qi deficiency of spleen and lung than the pediatric massage alone.

OBJECTIVE: To investigate the expression and cellular localization of nucleolin C23 during human umbilical vein endothelial cell (HUVEC) apoptosis induced by hydrogen peroxide (H(2)O(2)). METHODS: Apoptosis of HUVEC was induced by exposure to 0.5 mmol/L H(2)O(2) for different periods and detected by flow cytometry and activity of caspase-3. The mRNA and protein expression of nucleolin were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, respectively. The intracellular distribution of nucleolin was observed by indirect immunofluorescence. RESULTS: The percentage of apoptotic cells was increased significantly after treatment with H(2)O(2) for 12, 24 and 36 hours. The activity of caspase-3 reached the peak after treatment with H(2)O(2) for 4 h. RT-PCR showed that nucleolin C23 mRNA was decreased after 2, 4, and 8 hours treatment with H(2)O(2). Western blot showed that C23 protein level was decreased after 12 hours with an additional cleft band of 80 kD appeared after 8 hours. Density analysis showed that the 80 kD cleft band increased in a time-dependent manner. Immunofluorescence analysis demonstrated that H(2)O(2)-induced C23 redistribution from the nucleus to the cytoplasm. CONCLUSION H(2)O(2) could induce apoptosis accompanying with C23-cleavage and C23-translocation from the nucleus to the cytoplasm.
Apoptosis ; drug effects ; Cell Nucleus ; metabolism ; Cells, Cultured ; Cytoplasm ; metabolism ; Endothelial Cells ; cytology ; metabolism ; Humans ; Hydrogen Peroxide ; pharmacology ; Phosphoproteins ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; RNA-Binding Proteins ; biosynthesis ; genetics ; Umbilical Veins ; cytology

Chromogranin A ; metabolism ; Diagnosis, Differential ; Duodenal Neoplasms ; metabolism ; pathology ; surgery ; Ganglioneuroma ; metabolism ; pathology ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; Humans ; Male ; Middle Aged ; Neurofibroma ; metabolism ; pathology ; Paraganglioma ; metabolism ; pathology ; surgery ; Phosphopyruvate Hydratase ; metabolism ; S100 Proteins ; metabolism

Objective To observe the therapeutic effects of low frequency ultrasound enhanced thrombolysis (LFUET) on acute cerebral infarction (ACI) in rats.Methods The ACI animal models were established by injec- ting auto-thrombus into the rats' left middle cerebral arteries.They were then treated with urokinase,and received transcranial LFUET treatment at the same time.Nervous system functioning was assessed using NSS,and infarct vol- umes (IVs) were measured through tetrazolium chloride (TTC) staining.Results The NSS scores in the large- dose urokinase group (LDU group),the ultrasound plus small-dose urokinase group (USMU group) and in the in- farct group (Ⅰgroup) at 24 h after treatment were significantly lower than those before treatment.IVs in the two treat- ment groups are lower than those in theⅠgroup,but there was no significant difference between the LDU group and USMU group volumes.Conclusion LFUET can accelerate the recovery of nervous system function in rats after ACI,minimize IVs,and reduce the required dosage of urokinase.

OBJECTIVETo study the influence of the reference values for semen analysis proposed in the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen on the indication spectrum for intrauterine insemination (IUI).

METHODSWe retrospectively analyzed the clinical data of 111 cycles of IUI by the reference values for semen analysis in the 4th edition of the WHO Laboratory Manual (group A) and 84 cycles by the 5th edition (group B). We recorded and compared the percentages of various indications for IUI between the two groups.

RESULTSThe complications for IUI in groups A and B were as follows: asthenospermia (87.4% [97/111] vs 55.9% [47/84], P < 0.05), oligospermia (0 vs 0), teratospermia (51.4% [57/111] vs 35.7% [30/84]) , abnormal liquefaction (0.9% [1/111] vs O) , sexual dysfunction and genital malformation (0 vs 3.6% [3/84] , immune infertility (0.9% [ 1/111] vs O), and unexplained infertility (3.6% [4/111] vs 2. 4% [2/84 ] ). There were no significant differences between the two groups in the percentages of all the indications except that of asthenospermia.

CONCLUSIONThe reference values for semen analysis proposed in the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen have an evident influence on the indication spectrum for IUI by largely reducing the cases of IUI for male factors, prolonging the cycles of some patients, causing excessive diagnosis and treatment for females, and increasing their mental and economic burdens.

Adult ; Contraindications ; Female ; Humans ; Insemination, Artificial ; Male ; Pregnancy ; Reference Values ; Retrospective Studies ; Semen ; Semen Analysis ; World Health Organization

Epithelial-mesenchymal transition ( EMT ) is a process in which epithelial cells lose their morphology and function and gradually transformed into mesenchymal-like cells. It is considered that EMT is the main cause for tumor recurrence and metastasis .Many factors are involved in the regulation of EMT , such as E-cadherin , transforming growth factor-β, Wnt signaling pathway , microRNA and EMT-related transcription factors .This article reviews the research progress on EMT and the involved mechanisms , and thus to provide a new perspective on cancer therapy in the future .

BACKGROUND:Hand-foot-mouth disease has become a major public health issue in children in China. In the present prospective study we investigated the clinical characteristics and emergency management of children with severe encephalitis associated with NPE caused by enterovirus 71. METHODS:The study was conducted in 2 pediatric intensive care units (PICUs) over a 2-month period. Clinical records were reviewed of critically ill children with severe encephalitis associated with NPE caused by EV71 who were admitted to PICUs during the period of May to June 2008 in Fuyang. RESULTS:We reviewed the complete records of 36 children, of whom 23 (63.9％) were male and 13 (36.1％) female. Their age ranged from 4 to 48 months, with an average of 15.8 months. Al children except one were under 3 years of age. The overal mortality in these children was 19.4％. The average duration of critical life threatening signs and symptoms was 2.1 days (12 hours-5 days). Nervous system diseases included brainstem encephalitis in 27 children (75％), brainstem encephalitis associated with myelitis in 6 children (16.7％), and general encephalitis in 3 chidren (8.3％), respectively. In 12 patients of NPE (33.3％) pink or bloody bubble sputum and asymmetric pulmonary edema or hemorrhage was the primary manifestation but no typical exanthema was observed. Five children died of acute onset of NPE and / or pulmonary hemorrhage with rapid progression of cardiopulmonary failure within hours after admission. Therapeutic management consisted of mechanical ventilation and administration of mannitol, methylprednisolone, intravenous immunoglobulin (IVIG) and vasoactive drugs, associated with the need of fluid volume resuscitation in 9 (25％) of the 36 children. CONCLUSIONS:In children less than 3 years of age found to be affected by severe EV71 encephalitis associated with NPE, one fifth may die. The major organ systems infected by severe EV71 include the central nervous system, the respiratory system, and the cardiovascular system. Early diagnosis and evaluation, respiratory support, treatment of intracranial hypertension, and mainttenance of function of the cardiovascular system are the most important therapeutic measures.