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Objective To study the microscopic structure and morphological characteristics of Zebrafish eyeball and retina at different developmental stages, and to lay a foundation for visual research model. Methods Select eight groups of zebrafish at different ages, with six fish in each group, 48 fish in total. Optical microscopy and transmission electron microscopy were used to observe the eyeball structure of Zebrafish at different developmental stages, and the thickness of retinal each layer was measured to analyze the temporal and spatial development pattern. The morphological characteristics of various cells in the retina and the way of nerve connection were observed from the microscopic and ultrastructural aspects, especially the structural differences between rod cells and cone cells. Results The retina of Zebrafish can be divided into ten layers including retinal pigment epithelial layer, rod cells and cone cells layer, outer limiting membrane, outer nuclear layer, outer plexiform layer, inner nuclear layer, inner plexiform layer, ganglion cell layer, nerve fiber layer, inner limiting membrane. Rod cells had a smaller nucleus and a higher electron density than cone cells. Photoreceptor terminals were neatly arranged in the outer plexiform layer, forming neural connections with horizontal cells and bipolar cells, and several synaptic ribbons are clearly visible within them. In Zebrafish retina, ganglion cell layer and inner plexiform layer are the earliest developed. With the growth and development of Zebrafish, the thickness of rod cells and cone cells layer and retinal pigment epithelial layer gradually increases, and the retinal structure was basically developed in about 10 weeks. Conclusion The retinal structure of Zebrafish is typical, with obvious stratification and highly differentiated nerve cells. There are abundant neural connections in the outer plexiform layer. The ocular development characteristics of Zebrafish are similar to those of most mammals.
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italic>Schisandra chinensis is a traditional Chinese medicine with the functions of reinforcing deficiency, strengthening, and inducing astringency, appliable to treat the chronic cough and deficiency in breath, palpitation, and insomnia, etc. A hybrid mass spectrometry scanning strategy (high-definition data-independent/data-dependent acquisition, HDDIDDA), enabling the ion mobility separation and alternating data-independent acquisition/data-dependent acquisition, was established, which, in combination with in-house library-driven automatic peak annotation workflows facilitated by the UNIFI software, was utilized to systematically characterize the multi-classes of chemical components from S. chinensis. The use of an HSS T3 column (100 mm × 2.1 mm, 1.8 μm), 0.1% formic acid in H2O-acetonitrile as the mobile phase running at the flow rate of 0.3 mL·min-1, and column temperature at 35 ℃, could enable good separation of the S. chinensis components within 42 min. HDDIDDA scan in both the positive and negative ion modes was employed for data acquisition. Based on the automatic peak annotation, reference standards comparison, MS2 data interpretation, and literature analysis, we were able to identify or tentatively characterize 105 compounds in the S. chinensis decoction, involving 56 terpenoids, 42 lignans, five glycosides, one organic acid, and one flavonoid. HDDIDDA scanning can improve the coverage of data acquisition and improve the accuracy of identification, while CCS prediction analysis provides the possibility to distinguish isomers by the ion mobility technology. The results provide reference for the intelligent material basis research of TCM.
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This study was designed to comprehensively characterize and identify the chemical components in traditional Chinese medicine Psoraleae Fructus by establishing an ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF-MS) method in combination with in-house library. The chromatographic separation conditions(stationary phase, column temperature, mobile phase, and elution gradient) and key MS monitoring parameters(capillary voltage, nozzle voltage, and fragmentor) were sequentially optimized via single-factor experiments. A BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) was finally adopted, with the mobile phase consisting of 0.1% formic acid in water(A) and acetonitrile(B) at the flow rate of 0.4 mL·min~(-1) and column temperature of 30 ℃. Auto MS/MS was utilized for data acquisition in both positive and negative ion modes. By comparison with reference compounds, analysis of the MS~2 fragments, in-house library retrieval and literature research, 83 compounds were identified or tentatively characterized from Psoraleae Fructus, including 58 flavonoids, 11 coumarins, 4 terpenoid phenols, and 10 others. Sixteen of them were identified by comparison with reference compounds, and ten compounds may have not been reported from Psoraleae Fructus. This study achieved a rapid qualitative analysis on the chemical components in Psoraleae Fructus, which provided useful reference for elucidating its material basis and promoting the quality control.
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Chromatography, High Pressure Liquid , Medicine, Chinese Traditional , Tandem Mass Spectrometry , Cell Cycle , CoumarinsABSTRACT
OBJECTIVE@#To investigate the diagnostic efficacy of targeted biopsy (TBx), systematic biopsy (SBx), TBx+6-core SBx in prostate cancer (PCa) / clinically significant prostate cancer (cs-PCa) for patients with prostate imaging reporting and data system (PI-RADS) score of 5, and thereby to explore an optimal sampling scheme.@*METHODS@#The data of 585 patients who underwent multiparametric magnetic resonance imaging (mpMRI) with at least one lesion of PI-RADS score 5 at Peking University First Hospital from January 2019 to June 2022 were retrospectively analyzed. All patients underwent mpMRI / transrectal ultrasound (TRUS) cognitive guided biopsy (TBx+SBx). With the pathological results of combined biopsy as the gold standard, we compared the diagnostic efficacy of TBx only, SBx only, and TBx+6-core SBx for PCa/csPCa. The patients were grouped according to mpMRI T-stage (cT2, cT3, cT4) and the detection rates of different biopsy schemes for PCa/csPCa were compared using Cochran's Q and McNemar tests.@*RESULTS@#Among 585 patients with a PI-RADS score of 5, 560 (95.7%) were positive and 25(4.3%) were negative via TBx+SBx. After stratified according to mpMRI T-stage, 233 patients (39.8%) were found in cT2 stage, 214 patients (36.6%) in cT3 stage, and 138 patients (23.6%) in cT4 stage. There was no statistically significant difference in the detection rate of PCa/csPCa between TBx+6-core SBx and TBx+SBx (all P>0.999). Also, there was no statistically significant difference in the detection rate of PCa/csPCa between TBx and TBx+SBx in the cT2, cT3, and cT4 subgroups (PCa: P=0.203, P=0.250, P>0.999; csPCa: P=0.700, P=0.250, P>0.999). The missed diagnosis rate of SBx for PCa and csPCa was 2.1% (12/560) and 1.8% (10/549), and that of TBx for PCa and csPCa was 1.8% (10/560) and 1.4% (8/549), respectively. However, the detection rate of TBx+6-core SBx for PCa and csPCa was 100%. Compared with TBx+SBx, TBx and TBx+6-core SBx had a fewer number of cores and a higher detection rate per core (P < 0.001).@*CONCLUSION@#For patients with a PI-RADS score of 5, TBx and TBx+6-core SBx showed the same PCa/csPCa detection rates and a high detection rates per core as that of TBx+SBx, which can be considered as an optimal scheme for prostate biopsy.
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Male , Humans , Prostatic Neoplasms/pathology , Magnetic Resonance Imaging/methods , Retrospective Studies , Prostate/diagnostic imaging , Image-Guided Biopsy/methodsABSTRACT
Aim To evaluate the effects of Rutaecarpine(Rut)on the expression of SIRT1 and the senescence of vascular smooth muscle cells(VSMCs)induced by angiotensin Ⅱ.Methods VSMC senescencewas induced by exposure to AngⅡ(1 μmol·L-1)for 72 h.VSMCs were treated with different concentrations of Rut(0.3, 1, 3 μmol·L-1).TRPV1 competitive antagonist CAPZ(10 μmol·L-1)and AMPK inhibitor Compound C(1 μmol·L-1)were used to explore whether TRPV1/AMPK mediated the protective effect of Rut.The quantity of senescent cells were determined by senescence-associated SA-β-Gal staining, and the intracellular ROS level was measured by(DCFH-DA)fluorescent probe.The migration ability of VSMCs was evaluated by Wound-healing assay combined with Transwell assay.The protein level of longevity protein SIRT1 and senescence-related proteins p53, p21 and AMPK phosphorylation level were detected by Western blot.Results Rut significantly inhibited Ang Ⅱ-induced VSMC senescence and ROS production and prevented VSMCs migration.Preprocessing of TRPV1 antagonist CAPZ could abolish the protective effect of Rut.Ang Ⅱ inhibited the expression of longevity protein SIRT1.Rut recovered SIRT1 expression in a dose-dependent manner, while prevented the up-regulation of senescence-related proteins p53 and p21.Ang Ⅱ inhibited AMPK phosphorylation, pre-treatment with Rut restored AMPK phosphorylation level.CAPZ and Compound C eliminated the up-regulating function of Rut on SIRT1 expression.Conclusions Rut up-regulates the expression of SIRT1 and prevents the senescence and migration of VSMCs induced by Angiotensin Ⅱ, which is related to activation of the TRPV1/AMPK signaling pathway.
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Objective: To investigate the effects of IMPX977 on long term potentiation (LTP) at Schaffer collateral-CA1 synapses in vitro and on methyl CpG binding protein 2 (Mecp2) expression in mice cortex and hippocampus. Methods: Thirty-two C57BL/6 mice were randomly divided into four groups: control, olive oil (vehicle), IMPX977 low (5 mg/kg) and high (15 mg/kg) groups. Mice were administrated every other day orally for two weeks. Extracellular recording technique in vitro was used to record the effects of IMPX977 on Schaffer collateral-CA1 LTP pathway in acute mice hippocampal slices. The Mecp2 protein expression level was detected by Western blotting. Results: Compared to the control group, vehicle did not alter the synaptic transmission in Schaffer collateral-CA1 synapses, however, IMPX977 at concentrations of 5 mg/kg and 15 mg/kg significantly enhanced fEPSP (field excitatory postsynaptic potential) slope in Schaffer collateral-CA1 pathway to (179.6 ± 17.8)% and (191.4 ± 21.4)%, individually 60 min after HFS, IMPX977 improved LTP induction significantly at Schaffer collateral-CA1 pathway at least. Also, IMPX977 significantly elevated MeCP2 protein level in cortex. Conclusion: The effects of IMPX977 on synaptic transmission and Mecp2 protein expression provided convincing evidence that IMPX977 could be promising new drug candidates for Rett syndrome treatment.
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BACKGROUND@#Delivery room resuscitation assists preterm infants, especially extremely preterm infants (EPI) and extremely low birth weight infants (ELBWI), in breathing support, while it potentially exerts a negative impact on the lungs and outcomes of preterm infants. This study aimed to assess delivery room resuscitation and discharge outcomes of EPI and ELBWI in China.@*METHODS@#The clinical data of EPI (gestational age [GA] <28 weeks) and ELBWI (birth weight [BW] <1000 g), admitted within 72 h of birth in 33 neonatal intensive care units from five provinces and cities in North China between 2017 and 2018, were analyzed. The primary outcomes were delivery room resuscitation and risk factors for delivery room intubation (DRI). The secondary outcomes were survival rates, incidence of bronchopulmonary dysplasia (BPD), and risk factors for BPD.@*RESULTS@#A cohort of 952 preterm infants were enrolled. The incidence of DRI, chest compressions, and administration of epinephrine was 55.9% (532/952), 12.5% (119/952), and 7.0% (67/952), respectively. Multivariate analysis revealed that the risk factors for DRI were GA <28 weeks (odds ratio [OR], 3.147; 95% confidence interval [CI], 2.082-4.755), BW <1000 g (OR, 2.240; 95% CI, 1.606-3.125), and antepartum infection (OR, 1.429; 95% CI, 1.044-1.956). The survival rate was 65.9% (627/952) and was dependent on GA. The rate of BPD was 29.3% (181/627). Multivariate analysis showed that the risk factors for BPD were male (OR, 1.603; 95% CI, 1.061-2.424), DRI (OR, 2.094; 95% CI, 1.328-3.303), respiratory distress syndrome exposed to ≥2 doses of pulmonary surfactants (PS; OR, 2.700; 95% CI, 1.679-4.343), and mechanical ventilation ≥7 days (OR, 4.358; 95% CI, 2.777-6.837). However, a larger BW (OR, 0.998; 95% CI, 0.996-0.999), antenatal steroid (OR, 0.577; 95% CI, 0.379-0.880), and PS use in the delivery room (OR, 0.273; 95% CI, 0.160-0.467) were preventive factors for BPD (all P < 0.05).@*CONCLUSION@#Improving delivery room resuscitation and management of respiratory complications are imperative during early management of the health of EPI and ELBWI.
Subject(s)
Female , Humans , Infant , Infant, Newborn , Male , Pregnancy , Birth Weight , Bronchopulmonary Dysplasia , China/epidemiology , Delivery Rooms , Gestational Age , Infant, Extremely Low Birth Weight , Infant, Extremely PrematureABSTRACT
Objective: To explain the "multi-components, multi-targets, multi-pathways" mechanism of Erzhiwan in treating benign prostatic hyperplasia(BPH) based the network pharmacology. Method: Ingenuity pathway analysis(IPA) was used to construct components-targets-diseases network and PPI network, then the classified enrichment analysis of gene ontology (GO) and pathway enrichment analysis (KEGG) were carried out on the main function of its gene sets, so as to discuss the mechanism of Erzhiwan in the treatment of BPH. Result: Erzhiwan has 19 components in IPA; and apigenin,luteolin,oleanolic acid and quercitrin were common components of Ligustri Lucidi Fructus and Echiptae Herba and the main component of Erzhiwan. Muscarinic acetylcholine receptor M3 (CHRM3), muscarinic acetylcholine receptor M2 (CHRM2), urokinase plasminogen activator receptor (PLAUR), kinin releasing enzyme 3 (KLK3), cadherin 1 (CDH1), chemokines 3 (CCL3) and metalloproteinase-1 (MMP-1) were important targets for Erzhiwan to treat BPH. The target proteins in PPI network were enriched with 20 GO functions and 5 main KEGG pathways, and Docking was verified for relevant targets. Conclusion: Erzhiwan may play a role in treating BPH by activating MMP-1 and inhibiting KLK3 and CCL3 protein expressions, inducing apoptosis, inhibiting cell proliferation and intervening relevant pathways of mitogen-activated protein kinase/extracellular signal-regulated kinase(MAPK/ERK) and nuclear factor(NF)-κB(NF-κB).
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Background@#Structured reports are not widely used and thus most reports exist in the form of free text. The process of data extraction by experts is time-consuming and error-prone, whereas data extraction by natural language processing (NLP) is a potential solution that could improve diagnosis efficiency and accuracy. The purpose of this study was to evaluate an NLP program that determines American College of Radiology Breast Imaging Reporting and Data System (BI-RADS) descriptors and final assessment categories from breast magnetic resonance imaging (MRI) reports.@*Methods@#This cross-sectional study involved 2330 breast MRI reports in the electronic medical record from 2009 to 2017. We used 1635 reports for the creation of a revised BI-RADS MRI lexicon and synonyms lists as well as the iterative development of an NLP system. The remaining 695 reports that were not used for developing the system were used as an independent test set for the final evaluation of the NLP system. The recall and precision of an NLP algorithm to detect the revised BI-RADS MRI descriptors and BI-RADS categories from the free-text reports were evaluated against a standard reference of manual human review.@*Results@#There was a high level of agreement between two manual reviewers, with a κ value of 0.95. For all breast imaging reports, the NLP algorithm demonstrated a recall of 78.5% and a precision of 86.1% for correct identification of the revised BI-RADS MRI descriptors and the BI-RADS categories. NLP generated the total results in <1 s, whereas the manual reviewers averaged 3.38 and 3.23 min per report, respectively.@*Conclusions@#The NLP algorithm demonstrates high recall and precision for information extraction from free-text reports. This approach will help to narrow the gap between unstructured report text and structured data, which is needed in decision support and other applications.
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OBJECTIVE@#To evaluate the diagnostic performance of MRI for the assessment of inferior vena cava (IVC) wall invasion by IVC thrombus in patients with renal cell carcinoma (RCC).@*METHODS@#We retrospectively collected patients who underwent radical nephrectomy and thrombectomy for RCC between 2010 and 2018 at Peking University First Hospital. All the patients underwent imaging on a 1.5 Tesla or 3.0 Tesla MRI scanner. Fifty-six patients met the inclusion criteria. Preoperative imaging was reviewed by two radiologists blinded to details of the patient's surgical procedure and histopathology. Two radiologists measured the maximum anterior-posterior diameter and coronal diameters of the IVC and renal vein, and the craniocaudal extent of tumor thrombus, and evaluated the MRI features of IVC thrombus, including occlusion of the IVC lumen, the margin of the tumor thrombus (smooth vs. irregular), contact of the IVC thrombus and IVC wall, and altered signal of the IVC wall. Univariable and multivariable associations of clinical and radiographic features with IVC wall invasion were evaluated by Logistic regression.@*RESULTS@#Of the 56 patients [male: 43, female: 13, mean age: (55.64±0.43) years], 17 (30.36%) were detected with IVC wall invasion, and most were clear cell carcinoma. Tumor thrombus with IVC wall invasion showed an increase in length of IVC thrombus [(7.91±3.59) cm vs. (5.94±3.57) cm, P=0.049], and more features of complete occlusion of the IVC lumen (P=0.002), irregular margin of the IVC thrombs (P=0.005), contact of the IVC thrombus and IVC wall (P=0.001), and altered signal of the low-intensity vessel wall (P<0.001), with a sensitivity of 94.12% and a specificity of 79.49%.@*CONCLUSION@#The present study indicates that MRI could be a means of evaluating RCC with IVC wall invasion, and the combination of tumor thrombus length and subjective impression of IVC wall invasion achieved a high sensitivity and specificity for diagnosis.
Subject(s)
Female , Humans , Male , Middle Aged , Carcinoma, Renal Cell , Kidney Neoplasms , Magnetic Resonance Imaging , Nephrectomy , Retrospective Studies , Thrombectomy , Thrombosis , Vena Cava, InferiorABSTRACT
The intestinal absorption properties of four main effective components(gallic acid, ocinolglucoside, ethyl gallate and penta-O-galloyl-β-D-glucose) in Rhus chinensis extracts were investigated by in situ single-pass intestinal perfusion model in rats. The liquid accumulation of perfusion was corrected by gravimetry. The HPLC method was established to determine the concentration of the four effective components in the intestinal perfusion. It showed significant differences(Pethyl gallate>gallic acid>ocinolglucoside, with significant differences between them(P<0.05). In conclusion, gallic acid, orpheolglucoside, ethyl gallate and pentacyl-glucose could be absorbed in the whole intestine. Their absorption rate and permeation ability were related to the intestinal section and the perfusate concentration. These results indicated potential active transport or facilitated diffusion in the intestinal transport process of the four effective components.
Subject(s)
Animals , Rats , Chromatography, High Pressure Liquid , Hydroxybenzoates , Metabolism , Intestinal Absorption , Perfusion , Phytochemicals , Metabolism , Rhus , ChemistryABSTRACT
Extrusion-spheronisation method was used to prepare Rhus chinensis total phenolic acid pellets. The formula and preparation of R. chinensis total phenolic acid pellets were optimized. The formulas( drug loading capacity,diluent,wetting agent and anti-sticking agent) were determined by the single factor test with yield,appearance and performance as the indexes. The preparation was optimized by Box-Behnken design and response surface method,with the rate of extrusion,rate of spheronization and time of spheronization as the independent variables and the overall desirability value of yield,friability and roundness as the dependent variables. The optimal formula of pellets was as follows: drug loading capacity 28. 7%,MCC-lactose 9 ∶1,silicon dioxide as anti-sticking agent,and 60% ethanol as wetting agent. The optimal preparation was determined as follows: the rate of extrusion was 43 r·min-1,the rate of spheronization was 1 800 r·min-1,and the time of spheronization was 4 min. The absolute deviation between predicted value and estimated value under the conditions was less than 5. 0%,with a high degree of model fit. The preparation parameters obtained were accurate,reliable and reproducible. Under scanning electron microscopy( SEM),R. chinensis total phenolic acid pellets were uniform in diameter,round and smooth. The optimal formulation and process are stable and feasible for preparing R. chinensis total phenolic acid pellets.
Subject(s)
Drug Compounding , Methods , Hydroxybenzoates , Chemistry , Particle Size , Rhus , Chemistry , SolubilityABSTRACT
BACKGROUND@#Structured reports are not widely used and thus most reports exist in the form of free text. The process of data extraction by experts is time-consuming and error-prone, whereas data extraction by natural language processing (NLP) is a potential solution that could improve diagnosis efficiency and accuracy. The purpose of this study was to evaluate an NLP program that determines American College of Radiology Breast Imaging Reporting and Data System (BI-RADS) descriptors and final assessment categories from breast magnetic resonance imaging (MRI) reports.@*METHODS@#This cross-sectional study involved 2330 breast MRI reports in the electronic medical record from 2009 to 2017. We used 1635 reports for the creation of a revised BI-RADS MRI lexicon and synonyms lists as well as the iterative development of an NLP system. The remaining 695 reports that were not used for developing the system were used as an independent test set for the final evaluation of the NLP system. The recall and precision of an NLP algorithm to detect the revised BI-RADS MRI descriptors and BI-RADS categories from the free-text reports were evaluated against a standard reference of manual human review.@*RESULTS@#There was a high level of agreement between two manual reviewers, with a κ value of 0.95. For all breast imaging reports, the NLP algorithm demonstrated a recall of 78.5% and a precision of 86.1% for correct identification of the revised BI-RADS MRI descriptors and the BI-RADS categories. NLP generated the total results in <1 s, whereas the manual reviewers averaged 3.38 and 3.23 min per report, respectively.@*CONCLUSIONS@#The NLP algorithm demonstrates high recall and precision for information extraction from free-text reports. This approach will help to narrow the gap between unstructured report text and structured data, which is needed in decision support and other applications.
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We aimed to evaluate the feasibility of simultaneous image acquisition of multiple instantaneous switchable scan (MISS) for prostate magnetic resonance imaging (MRI) on 3T. Fifty-three patients were scanned with MRI due to suspected prostate cancer. Twenty-eight of them got histological results. First, two readers assessed the structure delineation and image quality based on images of conventional T2-weighted imaging (T2WI) and diffusion-weighted imaging (DWI) (CTD). Second, two readers identified the index lesion together, and then, reader one evaluated the contrast of index lesion on T2WI and signal ratio on apparent diffusion coefficient map. Third, they assigned Prostate Imaging Reporting and Data System (PI-RADS) score in consensus for the index lesion. After 4 weeks, the images of MISS were reviewed by the same readers following the same process. Finally, two readers gave preference for image interpretation, respectively. Kappa coefficient, Wilcoxon signed-rank test, paired-sample t-test, Bland-Altman analysis, and receiver operating characteristic (ROC) analysis were used for statistical analysis. The acquisition time of CTD was 6 min and 10 s, while the acquisition time of MISS was 4 min and 30 s. Interobserver agreements for image evaluation were κ = 0.65 and κ = 0.80 for CTD and MISS, respectively. MISS-T2WI showed better delineation for seminal vesicles than CTD-T2WI (reader 1: P < 0.001, reader 2: P = 0.001). The index lesion demonstrated higher contrast in MISS-T2WI (P < 0.001). The PI-RADS scores based on CTD and MISS exhibited high ability in predicting clinically significant cancer (area under curve [AUC] = 0.828 vs 0.854). Readers preferred to use MISS in 41.5%-47.2% of cases. MISS showed comparable performance to conventional technique with less acquisition time.
Subject(s)
Adult , Aged , Aged, 80 and over , Humans , Male , Middle Aged , Young Adult , Diffusion Magnetic Resonance Imaging/methods , Image Processing, Computer-Assisted/methods , Magnetic Resonance Imaging/methods , Prostate/diagnostic imaging , Prostatic Neoplasms/diagnostic imaging , Seminal Vesicles/diagnostic imagingABSTRACT
This article reviewed the relevant research on concept, substance, functions, and clinical application of Sanjiao during recent years. The study believed that Sanjiao is the "big viscera" integrating functions of viscus;"Sanjiao is invisible" means it couldn't be described; "Sanjiao is visible" theory has made certain achievements;Sanjiao is secondary channel for running of primordial qi; Qi transformation of lower-jiao is the foundation of blood, essence, bones and brain; The function of Sanjiao is the function of autonomic nervous system; The dysfunction of qi transformation and the disorder of qi activity of Sanjiao are generally deemed as pathogenesis of various diseases. This study also pointed out that researchers should pay more attention to physiological characteristics and functions of Sanjiao, with a purpose to help guide clinical practice.
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To study the application and effect of Role-playing in the teaching of Medical Immunology for foreign students.Role-playing method was tried using in the practical teaching of the pathway of complement activation and the related components in the"Medical Immunology" in Grade 2014(83 foreign students),and the teaching effect was analyzed and evaluated.In the study,we found that role-playing teaching method could not only help foreign students develop their initiative and comprehensively understand the difficulties of knowledge,but also cultivate their teamwork and interpersonal communication skills.At the same time,role-playing improved teaching efficiency and quality.Role-playing is a teaching method that meets the learning habits,characteristics,contents and rules of foreign students,which can improve the teaching quality and effect.
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This study was aimed to explore the effect of angiotensin converting enzyme 2 (ACE2) agonist diminazene aceturate (DIZE) on acute lung injury (ALI) induced by limb ischemia-reperfusion (LIR) in mice. Male 8-week-old wild-type and hACE2 transgenic ICR mice were randomly divided into 6 groups (6 in each group), including wild-type control (W), wild-type model (WL), wild-type model with DIZE administration (WLD), transgenic control (T), transgenic model (TL), and transgenic model with DIZE administration (TLD) groups. LIR model was established by 4 h reperfusion following 2 h ischemia of bilateral hindlimbs with rubber bands in mice. The WLD and TLD groups were pretreated with DIZE (15 mg/kg, i.p.) for 4 weeks before LIR. At the end of LIR, the mice were sacrificed and lung tissues were sampled. Indexes for evaluating lung injury include organ coefficient and wet/dry weight ratio (W/D), cell count and protein concentration of bronchoalveolar lavage fluid (BALF), as well as morphological change and pathological score were detected. Angiotensin II (Ang II) and Ang (1-7) levels in lung tissue were determined by using ELISA commercial kits. And the protein expressions of angiotensin II type 1 receptor (AT1) and Mas receptor protein in lung tissue were detected by Western blot. The results were as follows: (1) There was obvious lung injury in both the WL and TL groups. The lung injury in the TL group was lighter than that in the WL group. DIZE could attenuate the lung injury in both the two groups. (2) The WL group showed increased Ang II and decreased Ang (1-7) levels, whereas the TL group did not exhibit any changes of these two proteins. DIZE decreased the level of Ang II in both the WL and TL groups, and increased the level of Ang (1-7) in the WL group. (3) In the WL and TL groups, AT1 and Mas receptor protein expressions were up-regulated. DIZE reversed the change of AT1 protein expression, whereas further increased Mas receptor expression in both the two groups. These results suggest that DIZE may improve the renin-angiotensin system homeostasis by regulating ACE2-Ang (1-7)-Mas axis in local lung tissue and play a protective role in LIR-induced ALI in mice.
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Translating of scientific advances into clinical practice is a major challenge in the stroke research field in the past decades. There were many reasons involved:animal models might not accurately capture all aspects of clinical stroke in humans, the blind and randomized design principle was not closely followed, the inclusion and exclusion criteria was not previously established, sample size was inadequate, endpoint was not scientific nor blindly assessed, inadequate reporting of data and statistical flaws. To bridge the gap between experimental and clinical research, international consortia have attempted to establish standardized guidelines for study design and data report, which include optimizing animal models as well as experimental design, using innovative approaches to assess endpoint, making raw data and negative results available, establishing prior registration mechanism, conducting multicenter preclinical randomized controlled trials (pRCTs), systematic reviews and meta-analysis of preclinical studies, evolving the original focus on neuroprotection into a broader consideration of the role of neurovascular unit and ischemic cascade.
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Objective To establish and optimize the rat jugular vein catheterization model in our lab, and perform a crossover study using this model to compare the pharmacokinetic characters of a newly developed midazolam formulation to the existing preparation. Methods Six SD rats (half male and half female) received the right jugular vein catheterization when the rats were sufficiently anesthetized. One week after the operation, all the rats were used to conduct a cross-over double period pharmacokinetic study. Totally1.33 mg/kg midazolam solutions from automatic needle and clinic available injection were adminisered to the jugular vein catheterization rats via im route. The washout period was 5 days. Exact volume of blood samples at designed time points were taken through the catheter. After preparation, the concentrations of midazolam in rat plasma were determined by using established LC-MS/MS method. The corresponding pharmacokinetic parameters were calculated by WinNolin software. Results The rat jugular vein catheterization model was successfully built. Blood was easily sampled and rats were well tolerated, meeting the requirement of repeated blooding. This model solved the bottleneck of cross-over study performed in rats. The pharmacokinetic behavior of newly developed midazolam formulation had no difference with that of clinic injections. The relative bioavailability was around 99% . Conclusion Rat jugular vein catheterization model is proved to be that of a propagating technique to do the cross-over study and to evaluate the pharmacokinetic characters of novel formulations.
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DNA extraction is a basic technology of molecular biology. The purity and the integrality of DNA structure are necessary for different experiments of gene engineering. As commonly used materials in the clinical detection, the fast, efficient isolation and extraction of genomic DNA from peripheral blood mononuclear cells is very important for the inspection and analysis of clinical blood. At present, there are many methods for extracting DNA, such as phenol-chloroform method, salting out method, centrifugal adsorption column chromatography method (artificial methods), magnetic beads (semi-automatic method) and DNA extraction kit. In this article, a brief review of the principle for existing DNA blood extraction method, the specific steps and the assessment of the specific methods briefly are summarized.