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OBJECTIVE@#To observe the effects of electroacupuncture (EA) pretreatment on cardiac function, sympathetic nerve activity, indexes of myocardial injury and GABAA receptor in fastigial nucleus in rats with myocardial ischemia reperfusion injury (MIRI), and to explore the neuroregulatory mechanism of EA pretreatment in improving MIRI.@*METHODS@#A total of 60 male SD rats were randomly divided into a sham operation group, a model group, an EA group, an agonist group and an agonist+EA group, 12 rats in each group. The MIRI model was established by ligation of the left anterior descending coronary artery. EA was applied at bilateral "Shenmen" (HT 7) and "Tongli" (HT 5) in the EA group and the agonist+EA group, with continuous wave, in frequency of 2 Hz and intensity of 1 mA, 30 min each time, once a day for 7 consecutive days. After intervention, the MIRI model was established. In the agonist group, the muscone (agonist of GABAA receptor, 1 g/L) was injected in fastigial nucleus for 7 consecutive days before modeling, 150 μL each time, once a day. In the agonist+EA group, the muscone was injected in fastigial nucleus 30 min before EA intervention. The data of electrocardiogram was collected by PowerLab standard Ⅱ lead, and ST segment displacement and heart rate variability (HRV) were analyzed; the serum levels of norepinephrine (NE), creatine kinase isoenzyme MB (CK-MB) and cardiac troponin I (cTnI) were detected by ELISA; the myocardial infarction area was measured by TTC staining; the morphology of myocardial tissue was observed by HE staining; the positive expression and mRNA expression of GABAA receptor in fastigial nucleus were detected by immunohistochemistry and real-time PCR.@*RESULTS@#Compared with the sham operation group, in the model group, ST segment displacement and ratio of low frequency to high frequency (LF/HF) of HRV were increased (P<0.01), HRV frequency domain analysis showed enhanced sympathetic nerve excitability, the serum levels of NE, CK-MB and cTnI were increased (P<0.01), the percentage of myocardial infarction area was increased (P<0.01), myocardial fiber was broken and interstitial edema was serious, the positive expression and mRNA expression of GABAA receptor in fastigial nucleus were increased (P<0.01). Compared with the model group, in the EA group, ST segment displacement and LF/HF ratio were decreased (P<0.01), HRV frequency domain analysis showed reduced sympathetic nerve excitability, the serum levels of NE, CK-MB and cTnI were decreased (P<0.01), the percentage of myocardial infarction area was decreased (P<0.01), myocardial fiber breakage and interstitial edema were lightened, the positive expression and mRNA expression of GABAA receptor in fastigial nucleus were decreased (P<0.01). Compared with the EA group, in the agonist group and the agonist+EA group, ST segment displacement and LF/HF ratio were increased (P<0.01), HRV frequency domain analysis showed enhanced sympathetic nerve excitability, the serum levels of NE, CK-MB and cTnI were increased (P<0.01), the percentage of myocardial infarction area was increased (P<0.01), myocardial fiber breakage and interstitial edema were aggravated, the positive expression and mRNA expression of GABAA receptor in fastigial nucleus were increased (P<0.01).@*CONCLUSION@#EA pretreatment can improve the myocardial injury in MIRI rats, and its mechanism may be related to the inhibition of GABAA receptor expression in fastigial nucleus, thereby down-regulating the excitability of sympathetic nerve.
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Male , Animals , Rats , Rats, Sprague-Dawley , Cerebellar Nuclei , Electroacupuncture , Myocardial Reperfusion Injury/therapy , Receptors, GABA-A/genetics , RNA, MessengerABSTRACT
OBJECTIVE To analyze the main components of Chelidonii Herba-Corydalis Rhizoma （CHCR）， and to predict pharmacodynamic substances against estrogen receptor （ER） -positive breast cancer and their potential targets and signaling pathways， followed by verifying experiments. METHODS The ethanol extract of CHCR was analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）. The network pharmacology analysis was performed for the screened components. The network diagram of CHCR “active components-target-pathway” was constructed， and the enrichment pathway in vitro was validated. RESULTS A total of 58 chemical components were identified， including 57 alkaloids and 1 organic acid. A total of 38 active ingredients were screened from the network pharmacology， and 38 core targets were found in the protein-protein interaction network of “component-disease” intersection targets； 258 gene ontology entries and 137 Kyoto encyclopedia of genes and genomics pathways were obtained， mainly including estrogen signal pathway， phosphatidylinositol-3-kinase/protein kinase B （PI3K/Akt） signal pathway， etc. The results of validation test showed that the median inhibitory concentration of CHCR to MCF-7 cells was 693 μg/mL； 150， 300， 600 μg/mL CHCR could significantly reduce the expressions of phosphorylated PI3K， phosphorylated Akt， ERα protein and ESR1 mRNA （P＜0.01）. CONCLUSIONS The anti-ER-positive breast cancer effect of CHCR may be related to the regulation of ER and PI3K/Akt pathways， which has the characteristics of multi-component and multi-target effects.
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Objective To analyze the trend of death level and the probability of premature death caused by chronic obstructive pulmonary disease (COPD) in Jinshan District of Shanghai from 1980 to 2020, and to provide reference for the formulation of prevention and control measures of COPD. Methods The death cases of COPD in Jinshan District from 1980 to 2020 were collected through the death cause registration information system. The crude mortality, standardized mortality, age group mortality, early death probability and annual change percentage in different periods were calculated. Results From 1980 to 2020, the crude mortality of COPD in Jinshan District was 154.38/100 000, and the standardized mortality was 82.66/100,000. In the past 41 years, the standardized mortality of COPD showed a downward trend in males and females (APC=-1.79%, -2.52%, P<0.001). In the same period, the mortality of COPD in subjects aged 30-69 years old and subjects aged 70 years old and above also decreased (APC=-8.79%, -4.79%, P<0.001), and the probability of premature death caused by COPD showed a downward trend in males and females (APC=-9.61%, -10.71%, P<0.001). Conclusion The mortality rate and the probability of premature death of COPD in Jinshan District have decreased in the past 41 years. However, COPD is still one of the major chronic diseases that pose a serious threat to the health of residents in Jinshan District. Comprehensive prevention and treatment measures should be taken to reduce the mortality and the probability of premature death of COPD.
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Objective:Powders and decocted powders account for about 1/3 in the Catalogue of Ancient Famous Classical Formulas （the First Batch）， and have a very important position. Determination of preparation technology and particle size in the pulverization process is the key step in the research and development of powders and decocted powders following the original methods. However， there are many terms describing the preparation technology and particle size of powders and decocted powders in ancient Chinese medical books， and the parameters are not clear. Due to the lack of unified basis of particle size， the existing research results have not formed a uniform consensus. Based on ancient textual researches and experimental results， this article discusses the particle size of decocted powders and powders. Method:Through textual researches of the preparation technology and particle size of powders and decocted powders and powder classification in the 2020 edition of Chinese Pharmacopoeia， the specifications of pulverized particle size were suggested. In addition， Xiebaisan and Danggui Buxuetang were taken as examples to investigate the influence of different particle sizes （4， 10， 24 mesh） on the preparation process of decocted powders and the obtained decoction. Result:The particle size of 4 mesh was equivalent to that of ancient as big as hemp bean. The contents of index components in Xiebaisan and Danggui Buxuetang with particle size of 4 mesh were higher than that of 10 mesh and 24 mesh， but the particle size of 50 mesh was too fine to be filtered. Conclusion:The suggested particle sizes of powders and decocted powders are recommended as Cumo is the power through 10-mesh sieve， Mo is the power through 24-mesh sieve， Ximo is the power through 80-mesh sieve， as big as hemp bean is the power through 4-mesh sieve and not through 10-mesh sieve.
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Objective To analyze the trend of death level and the probability of premature death caused by gastric cancer in Jinshan District, Shanghai from 1980 to 2019, and to provide a basis for formulating prevention and control measures of gastric cancer. Methods The death cases of gastric cancer were collected through the death cause registration system in Jinshan District, and the crude gastric cancer death rate, standardized death rate, age group death rate, premature death probability and annual change percentage were calculated. Results From 1980 to 2019, the crude mortality of gastric cancer in Jinshan District was 22.38/100 000, and the standardized mortality was 13.85/100 000. Over these 40 years, the standardized mortality of gastric cancer showed a downward trend in both males and females (APC=-3.69%, -3.39%, P<0.001). In different periods, the mortality of gastric cancer in all age groups decreased. Over the past 40 years, the probability of premature death caused by gastric cancer showed a downward trend in both males and females (APC=-4.32%, -4.24%, P<0.001), but there was no significant downward trend in the probability of premature death caused by gastric cancer in males and females in the past 10 years. Conclusion Over the past 40 years, the mortality rate and the probability of premature death of gastric cancer in Jinshan District have shown a downward trend. However, gastric cancer is still one of the major malignant tumors that seriously threaten the health of residents in Jinshan District. Comprehensive prevention and treatment measures should be taken to reduce the mortality and the probability of premature death of gastric cancer.
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Objective:There were 92 kinds of compound preparations containing Ophiopogonis Radix in the 2015 edition of Chinese Pharmacopoeia， but there was no effective method to identify these compound preparations. Because Ophiopogonis Radix and Liriopes Radix are similar in appearance， it is easy to be confused in application. The aim of this study was to set up a thin layer chromatography （TLC） to identify compound preparations containing Ophiopogonis Radix and distinguish Ophiopogonis Radix and Liriopes Radix in the forms of decoction pieces and standard decoction. Method:In this study， decoction pieces of Ophiopogonis Radix and Liriopes Radix were collected and separately prepared as standard decoction. TLC was used to qualitatively identify decoction pieces and standard decoction of Ophiopogonis Radix and Liriopes Radix， and compound preparations containing Ophiopogonis Radix. In the TLC， the lower solution of chloroform-methanol-water （65∶35∶10） was selected as the developing agent and 10% sulfuric acid ethanol solution as the chromogenic agent. Result:The resolution of this TLC was good. Decoction pieces， standard decoction and preparations of Ophiopogonis Radix had the same characteristic strips， which were two bright white fluorescent strips under ultraviolet lamp （365 nm）. But these two characteristic strips were not existed in the TLC of decoction pieces and standard decoction of Liriopes Radix. The corresponding components of both of these two strips were identified as mixture containing saponins by LC-MSn， including ophiopogonin Ra， Tb， ophiopogonin D'， borneol glycoside， ophiopogonin C and Liriope muscari baily saponins C. Conclusion:The established TLC method， which has significant advantages such as high specificity and sensitivity， can be applied to the characteristic identification of decoction pieces and standard decoction of Ophiopogonis Radix， the identification of compound preparations containing Ophiopogonis Radix， and the distinction of Ophiopogonis Radix and Liriopes Radix， thus serving as an effective method to qualitatively identify Ophiopogonis Radix and its compound preparations.
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To study the compatibility rule of Simao Yongan Decoction,the rat single pass intestinal perfusion model in situ was used in this study. On the basis of early research,the five kinds of anti-inflammatory active ingredients,i.e. chlorogenic acid,liquiritin,hyperoside,angoroside C and isochlorogenic acid C in Simao Yongan Decoction were selected as research objects. The contents of the above five actives compounds with various compatibility combinations and in different intestinal segment perfusates were determined by using the method of ultra-performance liquid chromatography-mass spectrometry( UPLC-MSn). The kinetic parameters of intestinal absorption of the five anti-inflammatory active ingredients were calculated,which could be used to evaluate the intestinal absorption of each component in different combinations. The results showed that the absorption parameters of liquiritin in ileum were highest in Glycyrrhizae Radix et Rhizoma single herb,while the absorption parameters of other four components in ileum and duodenum were highest in the compatible combinations. Among them,the absorption parameters of chlorogenic acid in ileum and duodenum were highest in the whole prescription compatibility; ischlorogenic acid C showed higher absorption levels in the whole prescription and the herb compatibility of Lonicerae Japonicae Flos-Scrophulariae Radix-Glycyrrhizae Radix et Rhizoma. However,the absorption levels of hyperoside and angoroside C in different compatibilities were quite different in ileum and duodenum. In this study,the intestinal absorption of five anti-inflammatory active ingredients in Simiao Yongan Decoction with different compatibility combinations was investigated,revealing that the absorption of active ingredients varied with the different compatibility combinations and different intestinal segments. At the same time,the above research also indicated that the absorption of active ingredients could be obviously promoted by the compatibility of compound prescriptions,laying a foundation for the research on the compatibility rule of Simiao Yongan Detection from the biological point of view.
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Animals , Rats , Drugs, Chinese Herbal , Pharmacokinetics , Intestinal Absorption , Intestines , Phytochemicals , PharmacokineticsABSTRACT
Proper chromosome separation in both mitosis and meiosis depends on the correct connection between kinetochores of chromosomes and spindle microtubules.Kinetochore dysfunction can lead to unequal distribution of chromosomes during cell division and result in aneuploidy,thus kinetochores are critical for faithful segregation of chromosomes.Centromere protein A (CENP-A) is an important component of the inner kinetochore·plate.Multiple studies in mitosis have found that deficiencies in CENP-A could result in structural and functional changes of kinetochores,leading to abnormal chromosome segregation,aneuploidy and apoptosis in cells.Here we report the expression and function of CENP-A during mouse oocyte meiosis.Our study found that microinjection of CENP-A blocking antibody resulted in errors of homologous chromosome segregation and caused aneuploidy in eggs.Thus,our findings provide evidence that CENP-A is critical for the faithful chromosome segregation during mammalian oocyte meiosis.
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To explore the clinical and genetic characteristics of Williams-Beuren syndrome (WBS) and to raise awareness of the disease.The characteristics of clinical manifestations,personal history,car diac ultrasound,brain magnetic resonance imaging (MRI),electroencephalogram (EEG) and chromosome detection results of two cases with WBS were analyzed.The two patients were both male and the age was 11 months and 1 day,and 9 months and 9 days,respectively.They both suffered from cardiovascular malformation:case one presented supravalvular aortic stenosis,and case two showed atrial septal defect and patent ductus arteriosus.Both of the cases were exhibited characteristic facial features of WBS,including full orbital,spherical nose,fiat nasal bridge,long philtrum and thick lips.For the mental development,case one displayed moderate to severe developmental retardation,and case two showed severe developmental retardation.In addition,case one presented bilateral indirect inguinal hernia and hydrocele,and case two manifested feeding difficulties,buried penis and infantile spasms.Personal history:case one's mother had tocolytic therapy during pregnancy period,and case one was born at full-term by cesarean section due to amniotic fluid pollution.Supplementary examination:brain MRI of the two cases were no significant abnormalities;the EEG of case two showed hypsarrhythmia,and the epileptic spasms were recorded.Chromosome detection results:case one was identified as 7q11.23 deletion including the fragment deletion mutation of elastin (ELN) gene by multiplex ligation dependent probe amplification method,and case two was found with 7q11.21q11.23 deletion by high resolution G-band method.The two cases with WBS both had cardiovascular malformations,special facial features,mental retardation and connective tissue or urinary system abnormality.The supravalvular aortic stenosis of case one may be associated with the deletion of ELN gene,and the occurrence of epilepsy of case two may be related to the q11.21 deletion beyond the 7q11.23 region.
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<p><b>OBJECTIVE</b>To construct a eukaryotic expression plasmid containing human triggering receptor expressed on myeloid cells-1(TREM-1) gene.</p><p><b>METHODS</b>The entire gene coding region of human TREM-1 was amplified from total RNA of human peripheral blood by means of RT-PCR. The fragment of TREM-1 was cloned to vector pUCm-T. After digestion by restriction endonuclease BamH I and Pst I, the fragment was subcloned into the eukaryotic expressing vector pEGFP-C3. This recombinant vector was transfected into 293 cells using liposome. The expression level of TREM-1 was determined by fluorescence microscope and Western blot assay. The recombinant TREM-1 vector was transfected into THP-1 cells. After stimulation with 100 ng/ml LPS for 24 h, the mRNA levels of TNF-α and IL-1β were measured using RT-PCR.</p><p><b>RESULT</b>The expression vector was constructed, and the result of the DNA sequencing showed that the constructed plasmid containing the TREM-1 gene. Fluorescence microscope and Western blot analysis showed that TREM-1 protein was expressed in 293 cells successfully. After transfection into THP-1 cells, recombinant TREM-1 could upregulate the mRNA levels of TNF-α and IL-1β.</p><p><b>CONCLUSION</b>Eukaryotic expression plasmid pEGFP-TREM-1 is successfully constructed and showed biological activity.</p>
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Humans , Cells, Cultured , Genetic Vectors , Membrane Glycoproteins , Genetics , Plasmids , Genetics , Receptors, Immunologic , Genetics , Transfection , Triggering Receptor Expressed on Myeloid Cells-1ABSTRACT
Informed consent is the important thesis in life ethics and life law,which gives the patients and experimenters the right of self-decision making on medical procedures and experiments.As the development of public health and public health ethics,informed consent has been paid more attention,especially on informed consent problems in epidemiology and public health practice.The requirement of informed consent varies in different research fields.According to different practice purposes,the researchers choose relevant applicable methods.