Objective To study the determination method of total flavonoids in Gansu Astragali Radix and Hedysarum Polybotrys. Methods Calycosin glycosides etc. was selected as reference substances, comparison on the difference of absorption curves was done by ultraviolet spectroscopy and colorimetric method (NaNO2-Al(NO3)3-NaOH, AlCl3, Mg(Ac)2, NaOH, phosphomolybdic acid, HCl-Mg power). Results With colorimetric method, the maximum absorption wavelength of referrence and the test was inconsistent. The absorption peak shape was also different. With UV method, Calycosin glycosides in band Ⅱ (260 nm) showed a shoulder absorption. Astragali Radix and Hedysarum Polybotrys also showed characteristic shoulder absorptions in band Ⅱ with absorption wavelength at 263 nm and 265 nm. So the sample absorption wavelength is basically the same as that of the control sample. Conclusion Colorimetries usually used for determination of total flavonoids are not suitable for the comparison determination of Gansu Astragali Radix and Hedysarum Polybotrys. It is suitable for determining the contents of total flavonoids in samples by UV spectrophotometry at the band Ⅱ, which is the characteristic absorption band of isoflavone compound.

Objective To find a way to measure and count plane distribution of cells distributed on single layer and compare differences of undifferentiated mesenchymal cells of periosteum germinal layer from different parts of the body.Methods After counting the number of undifferentiated mesenchymal cells of periosteum germinal layer from different parts of the body microscopically and figuring out the number of cells per area unit in each periosteum specimen,the obtained data were statistically analyzed and the stratum structure of periosteum observed microscopically.Results The homogeneity of variance test showed homoscedasticity,with no statistical significance(P＞0.05).The analysis of variance found homoscedasticity but showed no statistical significance(F=0.253,P＞0.05).The periosteum of patel- la,tibial plateau and costa had two layers,while the periosteum of costal cartilage had three layers. Conclusions There is no conspicuous difference upon proliferation and evoluting activities of periosteum from different parts of body.Therefore,it is unnecessary to choose specific parts for drawing the periote- um in clinical situation.In the meantime,the structure of periosteum from different parts diversifies.

Abstract: Objective　To analyze the serotype distribution, drug resistance rate and drug resistance gene carrying of Streptococcus pneumoniae isolates in hospitalized patients, and evaluate the coverage of the vaccine to the serotype of Streptococcus pneumoniae in this area, so as to provide reference for the rational use of antibiotics in clinic. Methods　A total of 150 strains of non-repetitive Streptococcus pneumoniae isolated from inpatients from January 2015 to December 2019 were collected for serotyping and antimicrobial sensitivity test. The carrying rates of pbp2b, ermB and tetM were detected by PCR. Results　The PCR classification rate of 150 strains of Streptococcus pneumoniae was 93.1%, and the classification rate of capsular swelling test was 100%, and a total of 19 serotypes were divided, mainly 19F and 6B. Children's serotypes were predominantly 19F, 6B, and 15A; adult serotypes were predominantly 19F, 14, and 23F. The coverage rates of the PCV7, PCV10, PCV13 and PPV23 vaccines were 36.8%, 42.1%, 57.9% and 68.4%, respectively. Strains with serotypes of 19F, 6B, 3, and 23F had higher rates of resistance to antimicrobials. The sensitivity of Streptococcus pneumoniae to penicillin was greater than 96.0%. Antimicrobials with significant differences in resistance rates between invasive and non-invasive strains were penicillin, moxifloxacin, and levofloxacin. The percentage of strains carrying both ermB and tetM resistance genes was 96.0%, and the concordance rate between pbp2b, ermB and tetM resistance genes and the resistance phenotype was >98.0%. A total of 10 multi-resistance combinations were detected, with a multi-resistance rate of 62.6%, and the multi-drug resistance pattern of Streptococcus pneumoniae was mainly concentrated in the 19F and 6B serotypes. Conclusion　There are significant age differences in the serotypes of Streptococcus pneumoniae in this area. The vaccine currently used has low coverage in this region and therefore offer limited protection to the population. The drug resistance rates of Streptococcus pneumoniae varied significantly among serotypes. Erythromycin and tetracycline are not recommended for clinical treatment of Streptococcus pneumoniae. Penicillin can still be used as the first choice for clinical treatment of Streptococcus pneumoniae infection.

The study is aimed to ensure the quality and safety of medicinal plants by using ITS2 DNA barcode technology to identify Corydalis boweri, Meconopsis horridula and their close related species. The DNA of 13 herb samples including C. boweri and M. horridula from Lhasa of Tibet was extracted, ITS PCR were amplified and sequenced. Both assembled and web downloaded 71 ITS2 sequences were removed of 5. 8S and 28S. Multiple sequence alignment was completed and the intraspecific and interspecific genetic distances were calculated by MEGA 5.0, while the neighbor-joining phylogenetic trees were constructed. We also predicted the ITS2 secondary structure of C. boweri, M. horridula and their close related species. The results showed that ITS2 as DNA barcode was able to identify C. boweri, M. horridula as well as well as their close related species effectively. The established based on ITS2 barcode method provides the regular and safe detection technology for identification of C. boweri, M. horridula and their close related species, adulterants and counterfeits, in order to ensure their quality control, safe medication, reasonable development and utilization.
Base Sequence ; China ; Corydalis ; chemistry ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Papaveraceae ; chemistry ; classification ; genetics ; Phylogeny ; Plants, Medicinal ; chemistry ; classification ; genetics

OBJECTIVETo clone the p40 and p35 subunit cDNA of porcine IL-12(pIL-12) and construct the fusion gene of recombinant porcine single-chain interleukin-12 (pscIL-12).

METHODSThe total RNAs were extracted from porcine peripheral blood mononuclear cells (PBMCs) and porcine splenic lymphocytes for cloning pIL-12 p35 and p40 cDNA by RT-PCR. A hydrophobic polypeptide linker (Gly4Ser)3 was used for splicing two different gene fragments (pIL-12) p40+linker+p35 (pscIL-12) by recombinant PCR to construct pscIL-12 fusion gene. The pscIL-12 fusion gene was then inserted into pcDNA3.1(+) eukaryotic expression plasmid, and the resulted pcDNA3.1(+)-pscIL-12 was transfected into CHO-K1 cells via lipofectin. The expression of pscIL-12 mRNA in the transfected cells was identified by RT-PCR.

RESULTSThe sequence of the cloned porcine IL-12 p40 and p35 cDNA and the constructed pscIL-12 fusion gene were verified by PCR, restriction enzyme digestion and sequencing. The mRNA of pscIL-12 fusion gene was detected in the transfected CHO-K1 cells by RT-PCR.

CONCLUSIONThe constructed pcDNA3.1(+)-pscIL-12 eukaryotic expression plasmid allows expression of pscIL-12 in CHO-K1 cells, thus facilitating further study of the biological activity and adjuvant effect of pscIL-12 fusion protein.

Animals ; Base Sequence ; CHO Cells ; Cloning, Molecular ; Cricetinae ; DNA, Complementary ; genetics ; Interleukin-12 ; biosynthesis ; classification ; genetics ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Swine

BACKGROUNDGastric cancer (GC) is one of the most prevalent malignancies in the world today, with a high mortality rate. CDX2 is a Drosophila caudal-related homeobox transcription factor that plays an important role in GC. Phosphatase and tensin homologue deleted from chromosome 10 (PTEN) is an important tumor suppressor which is widely expressed in normal human tissues. The aim of the study was to determine the relationship and mechanism between CDX2 and PTEN in invasion and migration of GC cells.

METHODSpcDNA3-CDX2 plasmids were transfected into MGC-803 cells to up-regulate CDX2 protein, and small interfering RNA-CDX2 was transfected to down-regulate CDX2. The influence of CDX2 or PTEN on cell migration and invasion was measured by invasion, migration and wound healing assays. Western blotting assay and immunofluorescence were used to detect the expression of CDX2, PTEN, phosphorylation of Akt, E-cadherin and N-cadherin. Statistical significance was determined by one-way analysis of variance.

RESULTSThe results showed that CDX2 reduced the migration and invasion of GC cells (P < 0.05), and inhibited the activity of Akt through down-regulating PTEN expression (P < 0.05). CDX2 also restrained epithelial-mesenchymal transition of GC cells.

CONCLUSIONSCDX2 inhibited invasion and migration of GC cells by PTEN/Akt signaling pathway, and that may be used for potential therapeutic target.

CDX2 Transcription Factor ; Cell Line, Tumor ; Cell Movement ; genetics ; physiology ; Chromosomes, Human, Pair 10 ; genetics ; Epithelial-Mesenchymal Transition ; genetics ; physiology ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Microfilament Proteins ; genetics ; metabolism ; PTEN Phosphohydrolase ; genetics ; Phosphoric Monoester Hydrolases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Signal Transduction ; genetics ; physiology ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Tensins ; Wound Healing ; genetics ; physiology

Objective To explore the feasibility and methodology of radiofrequency catheter ablation (RFCA) guided by 3D navigation system (Ensite-NavX) for right atrioventricular accessory pathway.Method Thirty-three cases of right accessory pathway atrioventricular reentrant tachycardia including 16 cases in right free wall,3 in right middle septum,14 in right posterior septum; 23 cases of dominant accessory pathway and 10 cases of concealed were treated by RFCA guided by NavX navigation.NavX navigation modeling method or spatial localization method was exploited to locate target positioning.Result All patients were successfully ablated without serious complications.Among them,25 cases were operated without exposure to X-ray,7 patients were exposed for several seconds to verify catheter position,1 case in right free wall was ablated under X-ray combined with Swartz sheath ablation.Conclusion Nonfluoroscopy or less fluoroscopy RFCA for right atrioventricular accessory pathway with Ensite-NavX is safe and feasible,modeling or spatial orientation method are helpful to locate the ablation target positioning.

Objective To test the effects of TNF-α inhibitor Etanercept on myocardial ischemia/ reperfusion (MI/R) injury in posttraumatic mice,and explore related mechanisms.Methods Traumatic mouse model was established with Noble-Collip drum.Five days after trauma,Notch1 was knocked down by intramyocardial injection of Notch1 small interfering RNAs (siRNA) or scrambled siRNA (20 μg).Seven days after trauma,mice were subjected to MI/R (30 minutes ischemia followed by reperfusion).Sham operation was similarly performed without coronary artery ligation.Ten minutes before reperfusion,mice received Etanercept (8 mg/kg,i.p.).ELISA was used to detect plasma levels of TNF-α and troponin I (cTnI) and myocardial nitrotyrosine content.Twenty-four hours after reperfusion,left ventricular ejection fraction (LVEF) was measured by echocardiography.Infarct size was determined by Evans blue/2,3,5-triphenyl tetrazolium chloride (TTC) double staining.Cardiac caspase-3 activity was detected using a caspase-3 kit.Myocardial TNF-α and Notchl intracellular domains (Notch1 ICD) expressions were determined by Western blot.Chemiluminescence was used to assess myocardial superoxide anion content.Results (1) Compared to vehicle group,Etanercept treatment significantly reduced cTnl content,infarct size and caspase-3 activity (all P ＜0.01),while obviously increased LVEF (P ＜0.01).(2) Etanercept treatment also significantly reduced plasma and myocardial TNF-α contents (P ＜ 0.01),whereas markedly increased myocardial Notch1 ICD content (P ＜0.05).(3) Compared to scrambled siRNA group,Notch1 deficiency significantly increased cTnI content,infarct size and caspase-3 activity (P ＜ 0.05),whereas obviously reduced LVEF (P ＜ 0.05).(4) Etanercept significantly reduced myocardial superoxide anion and nitrotyrosine content (P ＜ 0.01),which was reversed by downregulation of Notch1 (P ＜ 0.05).Conclusions TNF-α inhibitor Etanercept can alleviate MI/R injury after trauma by reducing myocardial oxidative/nitrative stress via activating Notch1 signaling pathway.

OBJECTIVETo investigate the influence of intranasal instilling titanium dioxide (TiO2) nanoparticles on monoaminergic neurotransmitters at different-time exposure.

METHODSCD female mice were intranasally instilled three different-sized (25 nm, 80 nm and 155 nm) TiO, suspension every other day in a dose of 50 mg/kg body weight. The control group was instilled the same volume of Milli-Q water. Inductively coupled plasma-mass spectrometry (ICP-MS) was used to analyze the titanium contents in murine brain after exposure to TiO2 particles 2 days, 10 days, 20 days and 30 days. The monoaminergic neurotransmitters such as norepinephrine (NE), dopamine (DA), 5-hydroxytryptamine (5-HT), 5-hydroxyindole acetic acid (5-HIAA), 3, 4-dihydroxyphenylacetic acid (DOPAC), and homovanillic (HVA), were determined by reversed-phase high performance liquid chromatography (RP-HPLC) with electrochemical detector.

RESULTSAfter exposure to TiO, nanoparticles 10 days, the titanium contents in murine brain were increased, the titanium content in the 25 nm group was up to (1059.3 +/- 293.5) ng/g. In 20 days, the titanium content decreased slowly with the metabolism of titanium in vivo, but it kept at a high level, the content decreased to (654.7 +/- 269.2) ng/g in the 25 nm group. After exposure to TiO2 nanoparticles 30 days, the titanium contents had no obviously change. Because of the accumulation of TiO, in the brain, the contents of NE and 5-HT increased significantly after exposure to 80 nm and 155 nm TiO, nanoparticles 20 days, while the decreased contents of DA, DOPAC, HVA and 5-HIAA were observed.

CONCLUSIONThe inhaled TiO2 nanoparticles could be translocated to and deposited in murine brain after absorbing by nasal mucosa, and further influence the releases and metabolisms of monoaminergic neurotransmitters in brain.

Administration, Intranasal ; Animals ; Biogenic Monoamines ; metabolism ; Brain ; drug effects ; metabolism ; Brain Chemistry ; Female ; Metal Nanoparticles ; Mice ; Neurotransmitter Agents ; metabolism ; Time ; Titanium ; administration & dosage ; pharmacology

BACKGROUNDMucus hypersecretion in the respiratory tract and goblet cell metaplasia in the airway epithelium contribute to the morbidity and mortality associated with airway inflammatory diseases. This study aimed to examine the effect and mechanisms of simvastatin on airway mucus hypersecretion in rats treated with lipopolysaccharide (LPS).

METHODSMucus hypersecretion in rat airways was induced by intra-tracheal instillation of LPS. Rats treated with or without LPS were administered intra-peritoneally simvastatin (5 and 20 mg/kg) for 4 days. Expression of Muc5ac, RhoA and mitogen-activated protein kinases (MAPK) p38 in lung were detected by real-time polymerase chain reaction (PCR), immunohistochemistry or Western blotting. Tumor necrosis factor (TNF)-alpha and IL-8 in bronchoalveolar lavage fluid (BALF) were assayed by an enzyme-linked lectin assay and enzyme linked immunosorbent assay (ELISA).

RESULTSSimvastatin attenuated LPS-induced goblet cell hyperplasia in bronchial epithelium and Muc5ac hypersecretion at both the gene and protein levels in lung (P <0.05). Moreover, simvastatin inhibited neutrophil accumulation and the increased concentration of TNF-alpha and IL-8 in BALF follows LPS stimulation (P < 0.05). The higher dose of simvastatin was associated with a more significant reduction in Muc5ac mRNA expression, neutrophil accumulation and inflammatory cytokine release. Simultaneously, the increased expression of RhoA and p38 MAPK were observed in LPS-treated lung (P <0.05). Simvastatin inhibited the expression of RhoA and p38 phosphorylation in lung following LPS stimulation (P < 0.05). However, the increased expression of p38 protein in LPS-treated lung was not affected by simvastatin administration.

CONCLUSIONSSimvastatin attenuates airway mucus hypersecretion and pulmonary inflammatory damage induced by LPS. The inhibitory effect of simvastatin on airway mucus hypersecretion may be through, at least in part, the suppression of neutrophil accumulation and inflammatory cytokine release via inactivation of RhoA and p38 signaling pathway.

Animals ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; pharmacology ; Lipopolysaccharides ; toxicity ; Male ; Mucin 5AC ; secretion ; Rats ; Rats, Sprague-Dawley ; Respiratory Mucosa ; drug effects ; secretion ; Simvastatin ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; rhoA GTP-Binding Protein ; antagonists & inhibitors