OBJECTIVE: To observe the effect of electroacupuncture (EA) pretreatment of "biaoben acupoint combination" on cardiomyocyte mitochondrial fission in the rats with myocardial ischemia-reperfusion injury (MIRI) and explore its mechanism. METHODS: Fifty male SD rats were randomly divided into a sham-operation group, a model group, an EA pretreatment group, an EA pretreatment + Compound C group and an EA pretreatment+ML385 group, 10 rats in each group. In the EA pretreatment, the EA pretreatment + Compound C group and the EA pretreatment+ML385 group, EA was delivered at bilateral "Neiguan" (PC6), "Zusanli" (ST36) and "Guanyuan" (CV4) for 20 min, with continuous wave and 2 Hz of frequency, 1 mA of current, once daily for consecutive 7 days. On day 8, in the EA pretreatment + Compound C group and the EA pretreatment+ML385 group, 30 min before model preparation, the intraperitoneal injection with Compound C (0.3 mg/kg) and ML385 (30 mg/kg) was administered respectively. Except in the sham-operation group, the ligation of the left anterior descending coronary artery was performed to prepare MIRI rat model in the rest groups. In the sham-operation group, the thread was not ligated. After modeling, the content of reactive oxygen species (ROS) in the ischemic area was measured by flow cytometry, superoxide dismutase (SOD) was detected using xanthine oxidase method, and malondialdelyde (MDA) was detected using thiobarbituric acid (TBA) chromatometry. The morphology of myocardial tissue in the ischemic area was observed with HE staining, and the mitochondria ultrastructure of cardiomyocytes observed under transmission electron microscopy. Using immunofluorescence analysis, the positive expression of mitochondrial fission factor (MFF), mitochondrial fission 1 protein antibody (Fis1) and dynamin-related protein 1 (Drp1) was detected; and with immunohistochemical method used, the protein expression of adenosine monophosphate-activated protein kinase (AMPK), nuclear factor E2-associated factor2 (Nrf2) and Drp1 in the ischemic area was detected. RESULTS: Compared with the sham-operation group, the content of ROS and MDA in the myocardial tissue of the ischemic area, and the positive expression of MFF, Fis1 and Drp1 increased in the model group (P<0.01); the content of SOD and the protein expression of AMRK and Nrf2 decreased (P<0.01), and the protein expression of Drp1 elevated (P<0.01). Compared with the model group, the content of ROS and MDA in the myocardial tissue of the ischemic area, and the positive expression of MFF, Fis1 and Drp1 were dropped in the EA pretreatment group (P<0.01); the content of SOD and the protein expression of AMRK and Nrf2 rose (P<0.01), and the protein expression of Drp1 declined (P<0.01); and in the EA pretreatment+Compound C group and the EA pretreatment+ML385 group, the positive expression of MFF, Fis1 and Drp1, and the protein expression of Drp1 were all reduced (P<0.01). When compared with the EA pretreatment + Compound C group and the EA pretreatment+ML385 group, the content of ROS and MDA in the myocardial tissue of the ischemic area, and the positive expression of MFF, Fis1 and Drp1 were dropped in the EA pretreatment group (P<0.01); the content of SOD and the protein expression of AMRK and Nrf2 rose (P<0.01, P<0.05), and the protein expression of Drp1 decreased (P<0.05). In comparison with the model group, the EA pretreatment+Compound C group and the EA pretreatment+ML385 group, the cardiac muscle fiber rupture, cell swelling and mitochondrial disorders were obviously alleviated in the EA pretreatment group. The morphological changes were similar among the model group, the EA pretreatment+Compound C group and the EA pretreatment+ML385 group. CONCLUSION Electroacupuncture pretreatment of "biaoben acupoint combination" attenuates myocardial injury in MIRI rats, probably through promoting the phosphorylation of AMPK and Nrf2, inhibiting the excessive mitochondrial fission induced by Drp1, and reducing mitochondrial dysfunction caused by mitochondrial fragmentation and vacuolation.
Animals ; Electroacupuncture ; Male ; Rats, Sprague-Dawley ; Myocardial Reperfusion Injury/physiopathology* ; Myocytes, Cardiac/cytology* ; Rats ; Acupuncture Points ; Mitochondrial Dynamics ; Humans ; Reactive Oxygen Species/metabolism* ; NF-E2-Related Factor 2/genetics* ; Superoxide Dismutase/metabolism*

A high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) method was established for simultaneous determination of seven characteristic components from the active fraction of Alpiniae Officinarum Rhizoma in rat plasma, including galangin, kaempferol, kaempferide, pinocembrin, 1,7-diphenyl-4-en-3-heptanone, 5-hydroxy-7-(4-hydroxy-3-methoxyphenyl)-1-phenyl-3-heptanone(DHPA), and 7-(4-hydroxy-3-methoxyphenyl)-1-phenyl-4-en-3-heptanone(DPHB). The new developed HPLC-MS/MS method was applied to study the pharmacokinetics of the 7 characteristic components in rats with Helicobacter pylori gastritis. A Waters Sunfire C_(18) column(2.1 mm×150 mm, 3.5 μm) was used. The acetonitrile-aqueous solution(containing 0.1% formic acid) was adopted as the mobile phase for gradient elution. Seven components and internal standard(chlorogenic acid) were separated within 12 min. Mass spectrometric detection was performed in multiple reaction monitoring(MRM) mode using electrospray ionization(ESI) source with fast switching between positive and negative ions. The method was verified by specificity, linearity, precision, accuracy, recovery, matrix effect, and stability and met the requirements of pharmacokinetic study on the 7 components in rat plasma. Pharmacokinetic results showed that the average peak time(T_(max)) of the 7 components was 0.31-2.19 h, their elimination half-life(t_(1/2)) was 5.26-16.65 h, and the average residence time(MRT) was 6.29-31.03 h after the oral administration of the active fraction of Alpiniae Officinarum Rhizoma to rats with H. pylori gastritis. The plasma exposure levels of galangin and DHPA were higher than those of the other components. The concentration-time curves of four detected flavonoids showed obvious double peaks. This study elucidated the pharmacokinetic characteristics of 7 characteristic components from the active fraction of Alpiniae Officinarum Rhizoma in rats with H. pylori gastritis, providing a scientific basis for the identification of the pharmacodynamic substances of Alpiniae Officinarum Rhizoma for treatment of H. pylori gastritis and the clinical application of Alpiniae Officinarum Rhizoma in the prevention and treatment of H. pylori gastritis.
Animals ; Rats ; Chromatography, High Pressure Liquid/methods* ; Tandem Mass Spectrometry/methods* ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Helicobacter pylori/drug effects* ; Alpinia/chemistry* ; Rats, Sprague-Dawley ; Gastritis/metabolism* ; Helicobacter Infections/metabolism* ; Flavonoids/blood* ; Rhizome/chemistry* ; Liquid Chromatography-Mass Spectrometry

Humans ; SARS-CoV-2 ; COVID-19 ; Antibodies ; Antibodies, Viral/therapeutic use* ; Antibodies, Neutralizing/therapeutic use*

Objective To analyze the effect of angiopoietin-like protein 4(ANGPTL4) on M2 macrophage differentiation.Methods The frequency of M2 macrophages in spleen of ANGPTL 4-/-mice and the controls was detected by flow cytometry.And the changes of M2 macrophages was measured by LPS stimulation.F4/80 + macrophages was separated by flow cytometry and treated with LPS or recombinant ANGPTL 4 protein.The secretion of IL-4 in CD4 + T cells was observed after purified macrophages after co-culture with CD4 + T cells by the flow-cytometric intracellular staining method.Results There was no significant difference in the frequency of M2 macrophages in ANGPTL 4-/-mice and controls.LPS stimulation did not affect the expression of M2 macrophages from ANGPTL 4-/-mice.The macrophages from ANGPTL 4/-mice did not promote differentiation of CD4 + T cells into Th2 cells.After co-culturing of macrophages and CD4 + T cells for 48 h in vitro,IL-4 secretion of CD4 + T cells was not changed.Conclusions ANGPTL4 has no effects on M2 macrophage differentiation.

Objective To analyze the effect of angiopoietin-like protein 4(ANGPTL4) on M2 macrophage differentiation.Methods The frequency of M2 macrophages in spleen of ANGPTL 4-/-mice and the controls was detected by flow cytometry.And the changes of M2 macrophages was measured by LPS stimulation.F4/80 + macrophages was separated by flow cytometry and treated with LPS or recombinant ANGPTL 4 protein.The secretion of IL-4 in CD4 + T cells was observed after purified macrophages after co-culture with CD4 + T cells by the flow-cytometric intracellular staining method.Results There was no significant difference in the frequency of M2 macrophages in ANGPTL 4-/-mice and controls.LPS stimulation did not affect the expression of M2 macrophages from ANGPTL 4-/-mice.The macrophages from ANGPTL 4/-mice did not promote differentiation of CD4 + T cells into Th2 cells.After co-culturing of macrophages and CD4 + T cells for 48 h in vitro,IL-4 secretion of CD4 + T cells was not changed.Conclusions ANGPTL4 has no effects on M2 macrophage differentiation.

AIM To determine the contents of chemical consituents in Lycii Fructus from Qaidam Basin.METHODS Spectrophotometry was adopted in the content determination of polysaccharides,total flavonoids and carotenoid.HPLC was applied to the content determination of betaine and scopoletin.Kjeldahl method was used for the content determination of protein.Then principal component analysis was performed.RESULTS The contents of carotenoid,betaine and scopoletin in samples from six growing areas showed obvious differences (P ＜ 0.05),while those of polysaccharides,total flavonoids and protein exhibited no obvious differences (P ＞ 0.05).The contents of various constituents in samples at three picking time also had no obvious differences (P ＞ 0.05).The comprehensive score of principal components of samples from Delingha City was the highest,followed by that from Ulan County.CONCLUSION The quality of Lycii Fructus from Qaidam Basin from Delingha City is the best.

This paper reviews metabolomics/metabonomics technologies used in digestive disease clinical diagnosis, summarizes mainly two aspects for using metabolomic methods in current diagnosis for gastrointestinal disease, i.e. the one used for diagnosis of gastrointestinal malignancy, the other improving diagnosis of disease which can not be easily distinguished by traditional clinical methods. The future use of the metabonomics in clinic is also analyzed in the present paper.
Gastrointestinal Diseases ; diagnosis ; Humans ; Liver Function Tests ; methods ; Metabolome ; physiology ; Metabolomics ; methods ; Stomach Neoplasms ; diagnosis

AIM: To observe the different changes of neuroendocrine systems between the state of sport fatigue and poverty of movement. METHODS: 60 male Wistar rats were randomly divided into three groups: normal control group, sport fatigue model group and poverty of movement model group (20 rats in each group). The sport fatigue model was established by the method of combining basal diet and loaded swimming during 2 weeks, whereas the method of restricted activities was used to establish the poverty of movement model with total experimental time of 10 weeks. By the end of experiment, the climbing pole time was determined. The contents of hypothalamus thyrotropin releasing hormone (TRH), and serum norepinephrine (NE) and epinephrine (E) in rats with different treatments were determined by ELISA. In addition, the changes of hypothalamus corticotropin release hormone (CRH), pituitary adrenocorticotropic hormone (ACTH) and thyroid stimulating hormone (TSH), and serum corticosterone (CORT), triiodothyronine (T_3), tetraiodothyronine (T_4) were determined by radioimmunoassay to evaluate the functions of adrenergic nerve-adrenomedullin system, hypothalamo-pituitary-adrenal (HPA) axis and hypothalamo-pituitary-thyroid (HPT) axis. RESULTS: Compared to control group, the climbing pole time of the animals was obviously decreased in two model group. The adrenergic nerve-adrenomedullin system and HPA axis were inhibited in sport fatigue model rats, but HPT axis was unchanged. Interestingly, the HPA axis was hyperfunctional and HPT axis was inhibited in poverty of movement model rats. However, no change in the adrenergic nerve-adrenomedullin system was observed. CONCLUSION: Sport fatigue and poverty of movement all affect neuroendocrine system and lead to the adjustment mechanism imbalance, but the target and tendency are different.

OBJECTIVE:To establish a RP-HPLC method for the determination of aristolochic acid A in decoction of Caulis Aristolochiae Manshuriensis METHODS:The analytical column was Spherisorb ODS2 column(4 6mm?250mm,5?m) The mobile phase consisted of a mixture,methanol-water-acetic acid(70∶27∶1) The flow rate was 1 0ml/min The UV detection wavelength was 250nm RESULTS:The linear range was 0 0 128?g～0 4 096?g(r=0 9 999) The regression equation was Y=4 553 7+5 388 319 3X The average recovery of aristolochic acid A was 97 33%(RSD=2 34%) CONCLUSION:This method is simple,sensitive and accurate

Objective To provide applied anatomic data for relevant operations of blood vessels of perisacral promontory(BVPSP). Methods The composition of BVPSP including origin, course, diameter of the middle sacral vessels, the distance between the sacral promontory and the sacral 1 transverse trunk were observed on 37 adult cadavers. Result The BVPSP is composed of the common and internal iliac vessels, the superior segment of the middle sacral vessels and the sacral 1 transverse trunk. Middle sacral artery comes from abdominal aorta. Middle sacral veins are thin walled without valves. The average diameter of middle sacral artery and vein is 1.02 mm and 2.53 mm respectively. The distance between the sacral 1 transverse trunk and the sacral promontory is 5.75 mm. Conclusion The composition of BVPSP, especially middle sacral veins, plentiful vascular anastomosis are the anatomical basis leading to massive hemorrhage in the relevant operations.