Limit test of flavones in diterpene ginkgolides meglumine injection materials by UV-Vis and HPLC-DAD method was studied in this essay. The HPLC-DAD method has lower LOD (about 1% of the UV-Vis), that is, the sensitivity is higher than UV-Vis method. Through the analysis of the kinds of flavonoids ingredients in the samples by LC-MS, the three compounds with highest contents are kaempferol, quercetin and isorhamnetin. Kaempferol, quercetin and isorhamnetin were chosen as reference compounds for HPLC analysis, and the HPLC separation analysis was carried on an Agilent Eclipse plus C18 column (4.6 mm x 250 mm, 5 μm) with methanol and water containing 0.4% phosphoric acid (50: 50) as mobile phase, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm. This method has good specificity, precision and reproducibility. The LODs of quercetin, kaempferide and isorhamnetin were 27.6, 22.3, 29.5 μg x L(-1). The average recovery was 87.9% (RSD 3.3%), 91.7% (RSD 3.1%), 88.3 (RSD 1.3%) for quercetin, kaempferide and isorhamnetin, respectively. Based on the 10 batches of sample results and sensitivity of different HPLC, the content of total flavonoids ingredients of diterpene ginkgolides meglumine injection materials was limited no more than 2 x 10(-5). This method is simple, quick and has good maneuverability, and could be used to the limit test of flavonoids in the diterpene ginkgolides meglumine injection materials.
Chromatography, High Pressure Liquid ; methods ; Diterpenes ; analysis ; Drugs, Chinese Herbal ; analysis ; Flavones ; analysis ; Ginkgolides ; analysis ; Limit of Detection ; Mass Spectrometry ; methods

Objective To evaluate risks of symptomatic pulmonary embolism (PE) in deep vein thrombosis (DVT) patients complicating iliac vein compression syndrome (IVCS).Methods Between January 2010 and January 2015,patients diagnosed with lower extremity DVT and IVCS at our institution were included.The odds of symptomatic PE versus lower extremity DVT complicated with IVCS were assessed using Logistic regression models.Its association with age,gender,thrombophilic risk factors,degree of stenosis,filter implantation and symptomatic PE were assessed.Results Of 759 patients,410 patients had inferior vena cava (IVC) filters implanted and 349 had not.In filter placement group,3 patients suffered from symptomatic PE among 32 with thrombi intercepted in IVC filters.In the group with no IVC filters placement,39 cases suffered from symptomatic PE.Symptomatic PE was in positive correlation to respiratory disease and right lower extremity DVT,and negative correlation to IVC filter implantation.Conclusions In lower extremity DVT patients complicated with IVCS,the right lower extrenity DVT and respiratory disease are risk factors of symptomatic PE.IVC filter implantation prevents symptomatic PE from happening in most patients.

【Objective】 To study the differentiate expression genes and regulation signaling pathways related to the occurrence and development of dilated cardiomyopathy with heart failure, we used bioinformatics methods to explore gene chip in gene expression omnibus(GEO) and genes related to dilated cardiomyopathy in online mendelian inheritance in man(OMIM) . 【Methods】 GSE21610 and GSE29919 chip gene sets sequenced by Herz- &Diabeteszentrum NRW laboratory of the University of Bochum, Germany with myocardial biopsy specimens from clinical cases under the platform GPL570 and uploaded to GEO public database were selected to perform our study. Myocardial biopsy specimens from dilated cardiomyopathy with heart failure and normal cardiac function were regarded as the experimental group(14 and 21 cases, respectively) and the control group(12 and 8 cases, respectively). Up-regulated expression genes with the criteria: P < 0.05, were screened in GEO2R with the selected samples of two chips, which were then used to perform kyoto encyclopedia of genes and genomes(KEGG) pathways analysis, gene ontology(GO) function analysis, and protein-protein interaction(PPI) analysis, and the results were displayed through Volcano map, Venn map, Heat map, and Bubble charts with enrichment pathways drawn by R language packages. Meanwhile, KEGG pathways with the criteria: NOM. P < 0.05, and core genes relating to dilated cardiomyopathy with heart failure were performed through gene set enrichment analysis (GSEA). All candidate genes were then intersected with the reported genes in the OMIM, respectively, and clinical significance of these candidate genes was explored in relevant literatures. 【Results】 A total of 173 up-regulated expression genes with P < 0.05, were obtained by GEO2R, which are mainly related to inflammatory signals, cell proliferation and differentiation regulating, and classical apoptotic signaling pathways. These genes were intersected with the 754 reported genes in OMIM, and three reported up-regulated expression genes were obtained: NPPA for diagnosis of heart failure, APOA1 that associates with cytokine action, and COL6A1 that regulates lateral tubular remodeling. 158 and 46 core genes from KEGG pathways with NOM. P < 0.05 were obtained by GSEA, respectively, which were then intersected with the 754 reported genes in OMIM, and two core genes were obtained: PRKCA, enhancing myocardial contractility, and BMP2, promoting the development of heart failure. 【Conclusions】 Based on bioinformatics analysis, we found that the candidate genes PRKCA, BMP2, NPPA, and COL6A1 are likely to be closely related to the occurrence and development of dilated cardiomyopathy with heart failure, which can be used to reveal meaningful therapeutic clues and directions for the clinical treatment of dilated cardiomyopathy with advanced cardiac dysfunction.

ObjectiveTo analyze the risk factors for in-hospital mortality in patients with Takotsubo Syndrome （TTS）. MethodsHospitalization data of consecutive patients with TTS from February 2009 to January 2022 were retrospectively collected and analyzed. Patients were divided into survival group and death group according to outcomes. The basic clinical information， triggering factors， laboratory examinations， electrocardiogram， echocardiography， complications and treatments of the two groups were compared. Univariable logistic regression analysis was used to screen the possible risk factors for in-hospital mortality in TTS patients， and multivariable logistic regression analysis was used to determine the independent risk factors for in-hospital mortality in TTS patients. ResultsA total of 62 TTS patients were included in our study， including 21 males （33.9%）， 41 females （66.1%） and 26 postmenopausal women （41.9%）， with the mean age of （55.6±16.2） years， and physical triggers were found in 50 patients （80.6%）. 17 patients （27.4%） died while 45 patients （72.6%） survived during hospitalization. The death group had lower systolic blood pressure and left ventricular ejection fraction （LVEF）， higher incidence rate of syncope， higher level of N-terminal pro-B natriuretic peptide （NT-proBNP） and hypersensitive troponin T （hs TnT） when compared with survival group （all P value <0.05）. As for the triggering factors， the proportion of TTS induced by neurologic disorders in the death group was higher than that in the survival group （P<0.05）. The death group had higher rates of cardiogenic shock， malignant ventricular arrhythmia， atrial fibrillation， and respiratory failure （all P value <0.05）. Compared with the survival group， therapeutic dopamine， therapeutic norepinephrine， hemodialysis and mechanical ventilation were higher in the death group （all P value <0.05）. Univariable logistic regression analysis suggested that syncope， NT-proBNP， LVEF， neurologic disorders， cardiogenic shock， malignant ventricular arrhythmia， atrial fibrillation， respiratory failure， therapeutic dopamine， therapeutic norepinephrine， hemodialysis and mechanical ventilation were potential risk factors for in-hospital mortality in TTS patients （all P value <0.05）. Multivariable logistic regression analysis indicated that neurologic disorders ［OR（95%CI）=5.651（1.195，26.715），P=0.029］， atrial fibrillation ［OR（95%CI）=6.217（1.276，30.298）， P=0.024）］ and therapeutic norepinephrin ［OR（95%CI）=8.847（1.912，40.949）， P=0.005］ were independent risk factors for in-hospital mortality in TTS patients. ConclusionsNeurologic disorders， atrial fibrillation and therapeutic norepinephrin are independent risk factors for in-hospital mortality in patients with Takotsubo Syndrome. Clinically， attention should be paid to the prevention and treatment of neurologic disorders and atrial fibrillation； norepinephrine should be carefully used in patients with diagnosed TTS complicated with hemodynamic instability.

Objective To investigate the situations of close contacting with MDR-TB in family and analysis the necessity and urgency of family protection.Methods Nine typical primary MDR-TB cases in our hospital that close contacted with family pulmonary TB patients.their relatives contacted with them and their treatment prognosis were retrospectively analyzed.Results All of the 9 cases were transmitted by lineal MDR-TB relatives.In one case,3 generations were pumonary TB patients.All of 4 family members were pulmonary TBpatients in another case.3 cases were treated with operations:2 cases with pulmonary lobectomy and 1 case with intestinal resection.Treatment prognosis:3 cases cured,2 cases not cured,4 cases dead.Conclusions Family close contacting with MDR-TB patients should be intervened earlier:including health education,effective protection and health examination regularly et al.

Objective：Rat UREB1 protein coded by the gene UREB1 can specially bind to URE (upstream regulatory element) which is in the upstream of the promoter. It′ s reported that the protein of UREB1 promote the transcription of Dynorphin gene and inhibits p53 transactivation. This study was designed to clone human UREB1 gene and explore the relationship between UREB1 and the development of tumor. Methods： The artificial synthetic oligonucleotide was used as the probe to screen human brain cDNA library and human UREB1 gene was cloned. The antibody, which was produced using the recombinant UREB1 from E.coli as the antigen and immunizing the animals, was utilized for detecting the distribution of UREB1 in different tumor tissues. Results： The human UREB1 gene was cloned by using in situ hybridization for screening human brain cDNA library, and the nucleotide sequences and the deduced amino acid sequence of human UREB1 has 91％ homology with that of rat UREB1 identified previously. Western blot analysis revealed that the human UREB1 was present in all tumor tissues but the quantity of UREB1 in different tissues was not the same. Immunohistochemistry results shown that the human UREB1 distributes primarily in the cytoplasm and nuclear of tumor cells and nuclear UREB1 in carcinosarcoma is much more than that in adenoma. After analyzing the level of tyrosine phosphorylated UREB1 in a few tumor tissues, the result shown that the more malignant the tumor tissue was, the higher level the tyrosine phosphorylated of UREB1 was in that tumor tissues. Conclusion： Human UREB1 may be involved in the development of tumor and its tyrosine phosphorylation may affect the degree of tumor malignant.

OBJECTIVETo characterize the biological function of calmodulin (CaM) from Clonorchis sinensis (C. sinensis, Cs) and investigate its role in clonorchiasis-associated hepatic fibrosis.

METHODSThe full-length sequence of CsCaM gene was isolated from Cs cDNA library and its homologues were searched using BLASTx for comparison. Bioinformatics analysis was performed to compare the homologues and predict the physiochemical characteristics and functional domains. The gene was cloned in a prokaryotic plasmid and expressed in E. coli, and the recombinant protein was purified by affinity chromatography for immunizing rats to produce polyclonal antibodies, whose titer was determined using ELISA analysis. Immunoblotting analysis was carried out to determine of the purity and antibody recognition of CsCaM. Immunofluorescence assay was employed to analyze the tissue location of the protein. A rat model of liver fibrosis was established by introperitoneal injection of the recombinant protein.

RESULTSThe recombinant CsCaM protein obtained contained 150 amino acids with a theoretical molecular mass of 23.4 kD. CsCaM homologue had EF hand motifs. The recombinant pET-30a-CsCaM plasmid expressed in BL21 E. coli was about 23.4 kD. The total IgG antibody titer in the immunized mice reached the peak level (over 1: 51200) 2 to 4 weeks after the first injection. Immunohistochemistry showed that CsCaM located in the testis of adult C. sinensis. The rats receiving intraperitoneal injection of CsCaM showed severe liver inflammation with mild to moderate liver fibrosis.

CONCLUSIONThe pro-inflammation and pro-fibrosis effects of CsCaM in rat liver suggest its involvement in clonorchiasis- associated hepatic fibrosis.

Animals ; Antibodies, Helminth ; blood ; Antigens, Helminth ; immunology ; Calmodulin ; immunology ; Clonorchiasis ; immunology ; Clonorchis sinensis ; immunology ; Enzyme-Linked Immunosorbent Assay ; Gene Library ; Immunoglobulin G ; blood ; Inflammation ; Liver Cirrhosis ; parasitology ; Male ; Mice ; Rats ; Recombinant Proteins ; immunology

OBJECTIVETo investigate the change of Th17/Treg cell in patients with psoriasis arthritis (PA) and its clinical significance.

METHODSThe levels of IL-17 and TGF-β1 were measured by enzyme-linked immunosorbent assay(ELISA) in PA patients (n=35) and healthy controls(n=30). The frequencies of Th17 and Treg in the peripheral blood were detected by flow cytometry.

RESULTSCompared with the healthy controls, Th17/Treg in peripheral blood were significantly increased (p<0.05), Th17-related cytokine IL-17 significantly increased (p<0.05), and TGF-β1 significantly decreased (p<0.05) in the PA patients.

CONCLUSIONTh17/Treg cell and the related cytokines IL-17 and TGF-β1 may be involved in the pathogenesis of PA.

Aged ; Arthritis, Psoriatic ; blood ; immunology ; Case-Control Studies ; Female ; Humans ; Interleukin-17 ; blood ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology ; Transforming Growth Factor beta1 ; blood

OBJECTIVETo explore the mechanism of rehabilitation after middle cerebral artery occlusion (MCAO).

METHODSMCAO model was reproduced with two-kidney, two clip renovascular hypertensive rats stroke-prone (RHRSP), which were divided into two groups, the treated group (treated with electric stimulus) and the control group (untreated model) randomly. The rehabilitation of rats was evaluated by balance beam walking test. The ultrastructural changes of neurons and astrocytes, expressions of glial fibrillary acidic protein (GFAP)-positive cells, neurofilament (NF) protein, and cerebral capillary dilatation M-associated protein-2 (MAP2), as well as the neurons apoptosis and the number of dilatation of cerebral capillary in the margin of infarcted area were observed by the end of 1st, 3rd, 6th and 9th week after modeling.

RESULTSThe motor function of paralysed limbs recovered better in the treated group than that in the control group by the end of 3-9th week after MCAO, the expression of GFAP-positive cells in astrocytes and NF, MAP2 in neurons as well as the number of cerebral capillary dilatation at the margin of infarcted area were higher than those in the control group (P < 0.05).

CONCLUSIONElectric stimulation treatment could improve the recovery of motor function of paralyzed limbs. It might be due to the effect of electric stimulus in increasing astrocytes proliferation, reinforcing activity of neurons and evoking the dilatation of cerebral capillary.

Animals ; Apoptosis ; Astrocytes ; ultrastructure ; Electric Stimulation Therapy ; Glial Fibrillary Acidic Protein ; metabolism ; Infarction, Middle Cerebral Artery ; pathology ; physiopathology ; rehabilitation ; Male ; Neurons ; ultrastructure ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To observe the changes of electrocardiogram and serum cardiac troponin I at the early stage of severe crush injury in rats. METHODS: Crush injury was produced in Sprague-Dawley rats. The changes of electrocardiogram were recorded with the standard II, the serum levels of cardiac troponin I were studied by automated chemiluminescence assay. RESULTS: The ST segment elevated considerably after crush injury and lasted 24 h, the levels of serum cTnI were much higher than those of the control groupes after 6 h of injury. CONCLUSION Cardiomyocyte injury was induced in the early phase of crush injury.
Animals ; Crush Syndrome/physiopathology* ; Electrocardiography ; Extremities/injuries* ; Female ; Heart Injuries/physiopathology* ; Male ; Rats ; Rats, Sprague-Dawley ; Troponin I/blood*