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1.
Article in Chinese | WPRIM | ID: wpr-927347

ABSTRACT

OBJECTIVE@#To observe the clinical efficacy of acupoint injection combined with Vitalstim electrical stimulation for post-stroke dysphagia.@*METHODS@#A total of 98 patients with dysphagia after first stroke were randomized into an acupoint injection group (35 cases, 2 cases dropped off), an electrical stimulation group (31 cases, 3 cases dropped off) and a combination group (32 cases, 3 cases dropped off). Injection of mecobalamin into Tunyan point, Vitalstim electrical stimulation and the combination of injection of mecobalamin into Tunyan point and Vitalstim electrical stimulation were applied respectively in the 3 groups, once a day, 10 times as one course, 2 courses were required. Before and after treatment, the tongue muscle thickness and video fluoroscopic swallowing study (VFSS) score were observed in the 3 groups.@*RESULTS@#After treatment, the tongue muscle thickness was decreased (P<0.05), the VFSS scores were increased (P<0.05) compared with before treatment in the 3 groups, and the variation of tongue muscle thickness and VFSS score in the combination group was greater than the acupoint injection group and the electrical stimulation group (P<0.05).@*CONCLUSION@#Both acupoint injection of mecobalamin and Vitalstim electrical stimulation have therapeutic effect on dysphagia after stroke, and the two have synergistic effect.


Subject(s)
Acupuncture Points , Acupuncture Therapy , Deglutition , Deglutition Disorders/therapy , Electric Stimulation , Humans , Treatment Outcome
2.
Article in Chinese | WPRIM | ID: wpr-907747

ABSTRACT

Objective:To study the effect of levosimendan on coronary microembolization (CME)-induced myocardial injury and LOX-1/p38MAPK pathway.Methods:Microspheres were injected into coronary anterior descending branch to construct swine CME model, swine was given levosimendan by continuous intravenous drip for 24 h before modeling, and myocardial-specific overexpression of lectin-like oxidized low density lipoprotein receptor 1 (LOX-1) was achieved through coronary artery injection of adeno-associated virus (AAVs) at 2 weeks before modeling. Then, echocardiography was used to measure cardiac function; HE staining and HBFP staining were used to observe the pathological changes of myocardium and myocardial microinfarction area, respectively; ELISA was used to detect the serum level of cTnI; TUNLE staining was used to detect cardiomyocyte apoptotic index; the LOX-1, Bax, caspase-3 p12, Bcl-2, and p-p38 MAPK protein in myocardial tissue was observed by immunofluorescence method.Results:Compared to the sham group, the LVEF, LVFS, and CO value in the CME group were decreased, while the LVEDd value was increased significantly (all P<0.05); the area of myocardial micro-infarction, serum cTnI level and cardiomyocyte apoptotic rate in the CME group were increased significantly (all P<0.05); the protein levels of Bax, caspase-3 p12, LOX-1, and p-p38 MAPK were increased significantly, while the Bcl-2 level was decreased significantly ( P<0.05). Levosimendan pretreatment significantly improved cardiac dysfunction, reduced the area of myocardial micro-infarction and serum cTnI level, alleviated cardiomyocyte apoptosis, and significantly reduced the LOX-1 and p-p38 MAPK protein expression levels following CME (all P<0.05); while pretreatment with levosimendan and LOX-1 overexpression AAVs simultaneously abolished the effects of pretreatment with levosimendan alone (all P<0.05). Conclusion:Levosimendan alleviates CME-induced myocardial injury through inhibiting cardiomyocyte apoptosis mediated by LOX-1/p38 MAPK signaling pathway.

3.
Article in Chinese | WPRIM | ID: wpr-905134

ABSTRACT

Objective:To develop an automatic adjustment algorithm of bed height of multi-position lower limb rehabilitation robot, to meet the variety of leg lengths and training modes to avoid the collision between robot and ground. Methods:Six mathematical models of robot bed body height were established for six training modes of multi-position lower limb rehabilitation robot, which were described with leg length and bed tilt angle. The influence was analyzed that mechanical clearance and deflection as well as the jitter error of leg bracket during movement. Furthermore, a software related to these models was developed to automatically adjust the bed height for training. Volunteers were recruited to test actually. Results:The test data of bed height are consistent with the theoretical calculation of six mathematical models. Clearance and deflection did not affect the theoretical results of bed height. The end of robot's lower limb was always above the safe height during rehabilitation training. Conclusion:The automatic adjustment algorithm of bed height has been established, which can ensure that the rehabilitation robot runs at a safe height.

4.
Article in English | WPRIM | ID: wpr-903955

ABSTRACT

Coronary microembolization (CME) is associated with cardiomyocyte apoptosis and cardiac dysfunction. Puerarin confers protection against multiple cardiovascular diseases, but its effects and specific mechanisms on CME are not fully known. Hence, our study investigated whether puerarin pretreatment could alleviate cardiomyocyte apoptosis and improve cardiac function following CME. The molecular mechanism associated was also explored. A total of 48 Sprague-Dawley rats were randomly divided into CME, CME + Puerarin (CME + Pue), sham, and sham + Puerarin (sham + Pue) groups (with 12 rats per group). A CME model was established in CME and CME + Pue groups by injecting 42 μm microspheres into the left ventricle of rats. Rats in the CME + Pue and sham + Pue groups were intraperitoneally injected with puerarin at 120 mg/kg daily for 7 days before operation. Cardiac function, myocardial histopathology, and cardiomyocyte apoptosis index were determined via cardiac ultrasound, hematoxylin-eosin (H&E) and hematoxylin-basic fuchsin-picric acid (HBFP) stainings, and TdT-mediated dUTP nick-end labeling (TUNEL) staining, respectively. Western blotting was used to measure protein expression related to the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) pathway. We found that, puerarin significantly ameliorated cardiac dysfunction after CME, attenuated myocardial infarct size, and reduced myocardial apoptotic index. Besides, puerarin inhibited cardiomyocyte apoptosis, as revealed by decreased Bax and cleaved caspase-3, and up-regulated Bcl-2 and PI3K/Akt/GSK-3β pathway related proteins. Collectively, puerarin can inhibit cardiomyocyte apoptosis and thus attenuate myocardial injury caused by CME. Mechanistically, these effects may be achieved through activation of the PI3K/Akt/GSK-3β pathway.

5.
Article in English | WPRIM | ID: wpr-896251

ABSTRACT

Coronary microembolization (CME) is associated with cardiomyocyte apoptosis and cardiac dysfunction. Puerarin confers protection against multiple cardiovascular diseases, but its effects and specific mechanisms on CME are not fully known. Hence, our study investigated whether puerarin pretreatment could alleviate cardiomyocyte apoptosis and improve cardiac function following CME. The molecular mechanism associated was also explored. A total of 48 Sprague-Dawley rats were randomly divided into CME, CME + Puerarin (CME + Pue), sham, and sham + Puerarin (sham + Pue) groups (with 12 rats per group). A CME model was established in CME and CME + Pue groups by injecting 42 μm microspheres into the left ventricle of rats. Rats in the CME + Pue and sham + Pue groups were intraperitoneally injected with puerarin at 120 mg/kg daily for 7 days before operation. Cardiac function, myocardial histopathology, and cardiomyocyte apoptosis index were determined via cardiac ultrasound, hematoxylin-eosin (H&E) and hematoxylin-basic fuchsin-picric acid (HBFP) stainings, and TdT-mediated dUTP nick-end labeling (TUNEL) staining, respectively. Western blotting was used to measure protein expression related to the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) pathway. We found that, puerarin significantly ameliorated cardiac dysfunction after CME, attenuated myocardial infarct size, and reduced myocardial apoptotic index. Besides, puerarin inhibited cardiomyocyte apoptosis, as revealed by decreased Bax and cleaved caspase-3, and up-regulated Bcl-2 and PI3K/Akt/GSK-3β pathway related proteins. Collectively, puerarin can inhibit cardiomyocyte apoptosis and thus attenuate myocardial injury caused by CME. Mechanistically, these effects may be achieved through activation of the PI3K/Akt/GSK-3β pathway.

6.
Article in Chinese | WPRIM | ID: wpr-863706

ABSTRACT

Objective:To establish a method that could determine 7 chemical components including baicalin, berberine hydrochloride, aloe-emodin, rhein, emodin, chrysophanol and physcion in San huang tablet by high performance liquid chromatography-diode array detector (HPLC-DAD) method simultaneously with quantitative analysis method. Methods:The Diamonsil C18 column (4.6 mm × 250 mm, 5 μm) was adopted. The mobile phase was acetonitrile (A)-0.1% phosphoric acid solution (B) with gradient elution at a flow rate of 1.0 ml/min. The column temperature was set at 30 ℃. The detector was DAD. The detection wavelengths were 280 nm (baicalin), 265 nm (berberine hydrochloride), 254 nm (aloe-emodin, rhein, emodin, chrysophanol and physcion).Results:The linear ranges of baicalin, berberine hydrochloride, aloe-emodin, rhein, emodin, chrysophanol and physcion were within the ranges of 0.051 8-0.518 0 μg ( r=0.999 2), 0.024 6-0.245 9 μg ( r=0.999 8), 0.002 0- 0.020 2 μg ( r=0.999 6), 0.002 0-0.020 0 μg ( r=0.999 0), 0.002 0-0.020 1 μg ( r=0.999 9), 0.002 0-0.020 1 μg ( r=0.999 5), and 0.001 0-0.010 2 μg ( r=0.999 8), respectively. The average recoveries ( n=6) were 95.41%-100.59%, with RSDs less than 3.11%. Conclusions:This HPLC-DAD method is consistent with methodology and could simultaneously determine 7 chemical components in San huang tablet.

7.
Journal of Medical Postgraduates ; (12): 184-187, 2020.
Article in Chinese | WPRIM | ID: wpr-818399

ABSTRACT

ObjectiveAt present, there are many bedside tools for delirium, but these manual tools are time-consuming and poor feasible. The aim of this study was to establish a delirium screening scale, automatically extracting keywords from electronic medical records (EMR).MethodsWe selected electronic medical records of 779 elderly hospitalized patients in West China Hospital of Sichuan University from 2015 to 2017. Then, R software was used to automatically extract keywords to form a database undercritical ration, correlation coefficient and different analysis methods. Finally, the Delphi method and Analytic Hierarchy Process Weight were carried out to the construct weight coefficient, so as to form the formal scale.ResultsIn the study, we developed a formal scale consisting of 59 items and 11 dimensions. The score of the scale ranged from 0 to 53.4, with a mean value of 6.64, skewness of 2.6 and kurtosis of 8.2.ConclusionThe delirium screening scale based on the EMR can improve the recognition rate of delirium through intelligent and automatic warning, so as to early diagnosis and timely intervention of delirium.

8.
Chinese Critical Care Medicine ; (12): 750-755, 2019.
Article in Chinese | WPRIM | ID: wpr-754049

ABSTRACT

Objective To explore the effects of Hippo pathway on differentiation, proliferation, and migration of bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods BMSCs of C57BL/6 mice were identified using fluorescence-activated cellsorting analysis and the capabilities of osteogenic, chondrogenic and adipogenic differentiation were evaluated. The differentiation of BMSCs to typeⅡalveolar epithelial cells (AECⅡ) was induced by indirect co-culture with mouse lung epithelial cells (MLE-12) and small airway epithelial cell growth medium (SAGM). The Hippo pathway was regulated by 2-deoxy-D-glucose (2-DG) and 9E1, the effects of 2-DG and 9E1 on the expression of BMSCs surface proteins (SPB, SPC and SPD) mRNA and pro-SPC protein were detected by real time quantitative polymerase chain reaction (qRT-PCR) and Western Blot. The effect of Hippo pathway on differentiation of BMSCs to AECⅡ cells was evaluated. The effect of Hippo pathway on the proliferation of BMSCs was evaluated by methyl thiazolyl tetrazolium (MTT) assay (intervention of 0.1, 0.5, 1.0, 5.0 mmol/L 2-DG). The scratch test and Transwell chamber test were used to analyze the effect of Hippo pathway on migration ability of BMSCs to conditioned medium of acute respiratory distress syndrome (ARDS) lung tissue. Results 2-DG could activate Hippo pathway in a dose-dependent manner and promote the differentiation to AECⅡ and proliferation of BMSCs, the maximum effects were observed after 5 mmol/L of 2-DG treatment [SPB mRNA (2-ΔΔCT): 2.42±0.28 vs. 1.89±0.11, SPC mRNA (2-ΔΔCT): 8.06±0.68 vs. 6.59±0.79, SPD mRNA (2-ΔΔCT): 6.45±0.37 vs. 5.27±0.28, pro-SPC/β-actin: 5.80±1.86 vs. 4.93±1.18, proliferation rate:(145.46±18.18)% vs. (98.91±4.36)%, all P < 0.05], but 9E1 could reverse those effects through inhibition of Hippo pathway [SPB mRNA (2-ΔΔCT): 1.32±0.17 vs. 1.89±0.11, SPC mRNA (2-ΔΔCT): 3.91±0.34 vs. 6.59±0.79, SPD mRNA (2-ΔΔCT): 3.38±0.25 vs. 5.27±0.28, pro-SPC/β-actin: 2.48±0.17 vs. 4.93±1.18, proliferation rate: (80.00±7.27)% vs. (98.91±4.36)%, all P < 0.05]. The ability of horizontal migration [wound healing: (27.17±3.53)% vs. (52.45±6.52)%, P < 0.05] and homing BMSCs to conditioned medium of ARDS lung tissue [cell count (fold, relative to control): 2.77±0.21 vs. 1.90±0.19, P < 0.05] were increased after activation of Hippo pathway by 2-DG treatment, but those effects were reversed after inhibition of Hippo pathway by 9E1 treatment [wound healing: (79.89±8.42)% vs. (52.45±6.52)%, cell count (fold, relative to control): 1.69±0.13 vs. 1.90±0.19, both P < 0.05]. Conclusion Activation of Hippo pathway could enhance differentiation of BMSCs to AECⅡ, promote proliferation and ability of horizontal migration and homing BMSCs to conditioned medium of ARDS lung tissue in vitro.

9.
Chinese Critical Care Medicine ; (12): 281-287, 2019.
Article in Chinese | WPRIM | ID: wpr-753955

ABSTRACT

Objective To investigate the effects of Hippo signaling pathway on lung injury repair of mesenchymal stem cells (MSC) in acute respiratory distress syndrome (ARDS) and its mechanism. Methods Mouse bone marrow-derived MSC (mMSCs) cell lines with low expression of large tumor suppressor 2 (LATS2) were constructed by lentiviral vector transfection. Male C57BL/6 mice aging 6-8 weeks old were divided into four groups according to random number table (n = 36). The ARDS animal model (ARDS group) was reproduced by intratracheally injection of 2 g/L lipopolysaccharide (LPS) 50 μL, the normal saline (NS) control group was injected with an equal volume of NS. After 4 hours of model reproduction, 5×104 mMSCs transfected with blank lentivirus vector (MSC-shcontrol group) or shLATS2 lentivirus vector (MSC-shLATS2 group) were transplanted intratracheally, while NS control group and ARDS group were injected with equal volume of phosphate buffered saline (PBS). Mice were sacrificed at 3, 7, and 14 days after modeling, and lung tissue and bronchoalveolar lavage fluid (BALF) were harvested. Near-infrared fluorescence imaging, immunofluorescence staining and Western Blot were used to track mMSCs in lung tissue. Retension and proportion of mMSC differentiation into type Ⅱ alveolar epithelial cells (AECⅡ) were evaluated. Lung tissue wet weight/body weight ratio (LWW/BW) and total protein (TP) and albumin (ALB) in BALF were determined to reflect pulmonary edema. The expression of Occludin protein in lung epithelium was tested by Western Blot to reflect permeability of epithelium. The levels of interleukins (IL-1β, IL-6, IL-10) in BALF were assessed by enzyme-linked immunosorbent assay (ELISA) to reflect lung inflammation. Hematoxylin-eosin (HE) staining and modified Masson staining were carried out, and the scores were assessed to reflect lung injury and evaluate pulmonary fibrosis. Results The signal intensity of isolated lung fluorescence images at 3 days in the MSC-shLATS2 group was significantly higher than that in the MSC-shcontrol group (fluorescence intensity: 0.039±0.005 vs. 0.017±0.002, P < 0.05), the number of green fluorescent protein (GFP)-positive cells in lung tissue was also significantly higher than that in the MSC-shcontrol group (cells/HP:29.65±6.98 vs. 17.50±4.58, P < 0.05), but they all decreased with time; and the proportion of mMSCs differentiated into AECⅡ was significantly increased [(64.12±15.29)% vs. (19.64±3.71)%, P < 0.05]. Compared with the NS control group, the levels of surface active protein C (SPC) and Occludin protein in the ARDS group were significantly decreased, LWW/BW ratio and TP, ALB and inflammatory factors levels in BALF were significantly increased; extensive alveolar and interstitial edema, hemorrhage and diffuse inflammatory cell infiltration were found in lung tissue, and the lung injury score was significantly increased; collagen fibers were deposited in alveolar septum and alveolar cavity, and pulmonary fibrosis score was also increased significantly. Compared with the ARDS group, the expression levels of SPC and Occludin at 14 days in the MSC-shcontrol group and the MSC-shLATS2 group were significantly increased (SPC/β-actin: 0.51±0.12, 0.68±0.10 vs. 0.27±0.08, Occludin/β-actin: 0.49±0.19, 0.79±0.11 vs. 0.25±0.08, all P < 0.05), TP, ALB, IL-1β, IL-6 levels in BALF at 3 days were significantly decreased [TP (g/L): 8.08±1.72, 5.12±0.87 vs. 12.55±2.09; ALB (g/L): 0.71±0.21, 0.44±0.18 vs. 1.18±0.29, IL-1β(ng/L): 99.26±14.32, 60.11±8.58 vs. 161.86±25.17, IL-6 (ng/L): 145.54±13.29, 101.74±11.55 vs. 258.79±27.88, all P < 0.05], and IL-10 was significantly increased (ng/L: 190.83±22.61, 316.65±37.88, both P < 0.05). Furthermore, all the above parameters in the MSC-shLATS2 group were significantly improved as compared with those in the MSC-shcontrol group (all P < 0.05). LWW/BW ratio in the MSC-shLATS2 group was significantly lower than that in the ARDS group and the MSC-shcontrol group (mg/g: 9.85±1.51 vs. 16.78±1.92, 14.88±1.74, both P < 0.05). Conclusion Inhibiting Hippo signaling pathway by low expression of LATS2 could promote the retention of mMSCs in lung tissue and differentiation into AECⅡcells of ARDS mice, improve pulmonary edema and alveolar epithelial permeability, regulate pulmonary inflammatory response, and alleviate pathological damage and fibrosis of lung tissue.

10.
Article in Chinese | WPRIM | ID: wpr-751864

ABSTRACT

Objectives To determine the effect of Hippo signaling pathway on lung injury repair of bone-marrow derived mesenchymal stem cells (mMSCs) in murine lipopolysaccharide (LPS) induced acute respiratory distress syndrome (ARDS).Methods C57BL/6 mouse bone marrow-derived MSC (mMSCs) cell lines with low expression of large tumor suppressor 1 (LATS 1) were constructed by lentiviral vector transfection.ARDS was modeled by intratracheally injection of 2 mg/mL lipopolysaccharide (LPS)50 μL.C57BL/6 mice were randomly(random number) divided into four groups (n=36):normal control group,ARDS group,ARDS mice transplanted with mMSCs transfected with blank lentivirus vector (MSC-shcontrol group) or sh-LATS 1 lentivirus vector (MSC-shLATS 1 group).Mice were sacrificed at 3,7,and 14 d after modeling,and lung tissue and bronchoalveolar lavage fluid (BALF) were collected.Near-infrared fluorescence imaging,immunofluorescence staining and Western blot were used to evaluate retention and differentiation of mMSCs in lung tissue.Lung tissue wet weight/body weight ratio (LWW/BW) and total protein (TP) and albumin (ALB) in BALF were determined to reflect pulmonary edema.The expression of Occludin protein in lung epithelium was tested by Western blot.The levels of interleukins (IL-1β,IL-6,and IL-10) in BALF were assessed by enzyme-linked immunosorbent assay (ELISA).Lung injury score and pulmonary fibrosis score in lung tissue were assessed.Results The retention ofmMSCs at 3,7 and 14 d in the MSC-shLATS1 group were significantly higher than those in the MSC-shcontrol group [(26.25±4.58) vs (12.13±3.75) cells/HP,(20.49±3.86) vs (9.97±2.76) cells/HP,and (13.77±3.55) vs (6.89±2.10) cells/HP.all P<0.05],so was the differentiation of mMSCs into type Ⅱ alveolar epithelial cells at 14 d [(64.12±15.29)% vs (19.64±3.71)%,P<0.05].LWW/BW and TP and ALB in BALF at 3 and 14 d in the MSC-shLATS1 group [(9.85±1.51),(6.11±0.83) (mg/g) and (5.12±0.87),(3.05±0.87) (mg/mL)and (0.44±0.18),(0.33±0.04) (mg/mL)] were higher than those in the MSC-shcontrol group [(14.88± 1.74),(8.04±l.70)(mg/g) and (8.08±1.72),(5.94±1.20) (mg/mL) and (0.71±0.21),(1.07±0.29) (mg/mL)] (all P<0.05),so was the relative expression of Occludin protein[(0.79±0.11) vs (0.49±0).19),(P<0.05)].The levels of IL-1β and IL-6(pg/mL) in BALF in the MSC-shLATS1 group [(60.11±8.58),(101.74±11.55)] was lower than those in the MSC-shcontrol group [(99.26±14.32),(145.54±13.29)] (all P<0.05),but the levels of IL-10 in BALF in the MSC-shLATS 1 group (316.65±37.88)pg/mL was higher than those in the MSC-shcontrol group (190.83±22.61)pg/mL (P<0.05).Lung injury scores at 3 and 14 d in the MSC-shLATS1 group [(7.18±1.12),(3.33±0.49)] was lower than those in the MSC-shcontrol group [(9.72±1.45),(5.11±0.86)] (all P<0.05),so was pulmonary fibrosis score at 14 d [(0.68±0.12) vs (1.47±0.18),P<0.05].Conclusion Inhibition of Hippo signaling pathway through underexpression of LATS1 could improve the therapeutic effects of mMSCs in murine LPS-induced ARDS.

11.
Article in Chinese | WPRIM | ID: wpr-801547

ABSTRACT

Objective@#To optimize the method of simultaneous determination of four aflatoxins (B1, B2, G1 and G2) of ginger by the ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method and high-throughput method.@*Methods@#The aflatoxins were extracted from ginger by methanol-water (80:20, V/V) solution, concentrated and dried with nitrogen. The aflatoxins were detected by UPLC-MS/MS by using Waters Acquity UPLC BEH C18 chromatographic column. The mobile phase was 0.1% formic acid water (A phase) -0.1% formic acid methanol (B phase), gradient elution, flow rate 0.35 ml/min, mass spectrometry was electrospray ion source, positive ion scanning mode, multi reaction ion monitoring were using.@*Results@#Quantification of four aflatoxins by matrix matching standard curve. The linear was good in the range of 0.125-20.000 ng/ml, and the correlation coefficients were all greater than 0.999 0. The ginger sample detection was 0.125-0.300 μg/kg and 0.125-1.000 μg/kg, respectively. The average recoveries were 81.7%-96.0%, and the relative standard deviation (RSD) was lower than 7.53%.@*Conclusions@#This method is simple, rapid, sensitive and low limit of detection, which can meet the requirements for the detection of trace aflatoxins residues in ginger.

12.
Chinese Critical Care Medicine ; (12): 1143-1148, 2019.
Article in Chinese | WPRIM | ID: wpr-797535

ABSTRACT

Objective@#To investigate the regulatory effect of Hippo signaling pathway mediated by large tumor suppressor gene 2 (LATS2) on biological behavior of mice bone marrow mesenchymal stem cells (BMSCs) in vitro.@*Methods@#BMSCs of C57BL/6 mice were cultured in vitro and passed to the 6th to 7th generations for experiment. BMSCs with activated and inactivated LATS2 were constructed with lentiviral vectors transfections. The BMSCs were allocated to blank control group (MSC group), empty vector control group (MSC-eGFP group), LATS2-overexpressing group (MSC-LATS2 group), empty vector without LATS2 shRNA control group (MSC-shcontrol group) and LATS2-underexpressing group (MSC-shLATS2 group). The transduction efficiencies mediated by the lentiviral vectors were evaluated by flow cytometry. The mRNA and protein expressions of LATS2, phosphorylation of Yes associated protein (p-YAP) and 14-3-3 were quantified by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot respectively. Osteogenic and adipogenic differentiation of BMSCs were evaluated through alizarin red and oil red O staining. Proliferation of BMSCs was evaluated using the CCK-8 assay. The effect of Hippo pathway on horizontal and vertical migration ability of BMSCs was measured by the scratch test and Transwell chamber test.@*Results@#The transduction efficiencies mediated by the lentiviral vectors were 93.1%-97.1%. Compared with MSC-eGFP group, the expressions of LATS2, p-YAP and 14-3-3 in MSC-LATS2 group were significantly elevated [LATS2 mRNA (2-ΔΔCT): 2.55±0.13 vs. 1.08±0.05, LATS2/GAPDH: 2.63±0.11 vs. 1.06±0.08, p-YAP/total YAP: 1.67±0.11 vs. 1.00±0.04, 14-3-3/β-actin: 2.22±0.20 vs. 0.98±0.06, all P < 0.05], however, compared with MSC-shcontrol group, the expressions in MSC-shLATS2 group were significantly reduced [LATS2 mRNA (2-ΔΔCT): 0.10±0.01 vs. 1.01±0.05, LATS2/GAPDH: 0.09±0.01 vs. 1.05±0.06, p-YAP/total YAP: 0.10±0.02 vs. 1.10±0.09, 14-3-3/β-actin: 0.05±0.01 vs. 0.90±0.08, all P < 0.05]. It suggested that high and low expression of LATS2 could activate or inhibit Hippo pathway. The osteogenic and adipogenic differentiation and proliferation rate of BMSCs in MSC-LATS2 group were significantly lower than those in MSC-eGFP group, however, those in MSC-shLATS2 group were significantly higher than MSC-shontrol group (all P < 0.05). It suggested that high and low expression of LATS2 could inhibit or promote osteogenesis, adipogenesis and cell proliferation of BMSCs. Scratch test and Transwell chamber test showed that the degree of scratch healing in MSC-LATS2 group was significantly lower than that in MSC-eGFP group [(22.11±3.02)% vs. (45.99±6.58)%], while the number of cells migrating to the subventricular layer of Transwell was significantly reduced (cells/MP: 20.82±3.05 vs. 111.33±13.28, both P < 0.05); the degree of scratch healing in MSC-shLATS2 group was significantly higher than that in MSC-shcontrol group [(70.32±7.17)% vs. (39.28±2.98)%], the number of cells migrating to the subventricular layer of Transwell was increased significantly (cells/MP: 206.19±30.58 vs. 120.10±25.10, both P < 0.05). It suggested that high and low expression of LATS2 could inhibit or promote the horizontal and vertical migration of BMSCs.@*Conclusion@#LATS2-mediated alteration of Hippo pathway could modulate differentiation, proliferation, and migration of mesenchymal stem cells in vitro.

13.
Chinese Critical Care Medicine ; (12): 1143-1148, 2019.
Article in Chinese | WPRIM | ID: wpr-791040

ABSTRACT

Objective To investigate the regulatory effect of Hippo signaling pathway mediated by large tumor suppressor gene 2 (LATS2) on biological behavior of mice bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods BMSCs of C57BL/6 mice were cultured in vitro and passed to the 6th to 7th generations for experiment. BMSCs with activated and inactivated LATS2 were constructed with lentiviral vectors transfections. The BMSCs were allocated to blank control group (MSC group), empty vector control group (MSC-eGFP group), LATS2-overexpressing group (MSC-LATS2 group), empty vector without LATS2 shRNA control group (MSC-shcontrol group) and LATS2-underexpressing group (MSC-shLATS2 group). The transduction efficiencies mediated by the lentiviral vectors were evaluated by flow cytometry. The mRNA and protein expressions of LATS2, phosphorylation of Yes associated protein (p-YAP) and 14-3-3 were quantified by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot respectively. Osteogenic and adipogenic differentiation of BMSCs were evaluated through alizarin red and oil red O staining. Proliferation of BMSCs was evaluated using the CCK-8 assay. The effect of Hippo pathway on horizontal and vertical migration ability of BMSCs was measured by the scratch test and Transwell chamber test. Results The transduction efficiencies mediated by the lentiviral vectors were 93.1%-97.1%. Compared with MSC-eGFP group, the expressions of LATS2, p-YAP and 14-3-3 in MSC-LATS2 group were significantly elevated [LATS2 mRNA (2-ΔΔCT): 2.55±0.13 vs. 1.08±0.05, LATS2/GAPDH: 2.63±0.11 vs. 1.06±0.08, p-YAP/total YAP: 1.67±0.11 vs. 1.00±0.04, 14-3-3/β-actin: 2.22±0.20 vs. 0.98±0.06, all P < 0.05], however, compared with MSC-shcontrol group, the expressions in MSC-shLATS2 group were significantly reduced [LATS2 mRNA (2-ΔΔCT): 0.10±0.01 vs. 1.01±0.05, LATS2/GAPDH: 0.09±0.01 vs. 1.05±0.06, p-YAP/total YAP: 0.10±0.02 vs. 1.10±0.09, 14-3-3/β-actin: 0.05±0.01 vs. 0.90±0.08, all P < 0.05]. It suggested that high and low expression of LATS2 could activate or inhibit Hippo pathway. The osteogenic and adipogenic differentiation and proliferation rate of BMSCs in MSC-LATS2 group were significantly lower than those in MSC-eGFP group, however, those in MSC-shLATS2 group were significantly higher than MSC-shontrol group (all P < 0.05). It suggested that high and low expression of LATS2 could inhibit or promote osteogenesis, adipogenesis and cell proliferation of BMSCs. Scratch test and Transwell chamber test showed that the degree of scratch healing in MSC-LATS2 group was significantly lower than that in MSC-eGFP group [(22.11±3.02)% vs. (45.99±6.58)%], while the number of cells migrating to the subventricular layer of Transwell was significantly reduced (cells/MP:20.82±3.05 vs. 111.33±13.28, both P < 0.05); the degree of scratch healing in MSC-shLATS2 group was significantly higher than that in MSC-shcontrol group [(70.32±7.17)% vs. (39.28±2.98)%], the number of cells migrating to the subventricular layer of Transwell was increased significantly (cells/MP: 206.19±30.58 vs. 120.10±25.10, both P < 0.05). It suggested that high and low expression of LATS2 could inhibit or promote the horizontal and vertical migration of BMSCs. Conclusion LATS2-mediated alteration of Hippo pathway could modulate differentiation, proliferation, and migration of mesenchymal stem cells in vitro.

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Article in Chinese | WPRIM | ID: wpr-707114

ABSTRACT

This article discussed the possible physiological mechanism of "treating waist by abdominal massage therapy" from the perspectives of TCM and modern medicine, and analyzed the effects of abdominal massage from the nerve, fascia, visceral, muscle and other aspects, with a purpose to elucidate that the occurrence of relevant diseases may be caused by a variety of factors, and different massage techniques have different effects on the body.

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Article in Chinese | WPRIM | ID: wpr-743199

ABSTRACT

Objective To investigate the role of nicorandil pretreatment on protecting myocardium after coronary microembolization (CME) and on the PDCD4/NF-κB/TNF-α signaling pathway in miniature pigs. Methods Fifteen Bama miniature pigs were randomly(random number) divided into the sham operation group (sham group), microembolization group (CME group) and CME plus nicorandil group, with 5 pigs in each group. The CME model was constructed by injecting polyethylene microspheres via microcatheter into the left anterior descending artery, and pigs in the sham group were injected with the same amount of saline. Pigs in the CME plus nicorandil group were injected intravenously with nicorandil (150 μg/kg) via ear vein 30 min before CME. Cardiac function indexes were measured using cardiac ultrasonography. The expression of PDCD4 and TNF-α mRNA in myocardium were detected by fluorescence quantitative PCR, and the protein expression of PDCD4 and TNF-α in myocardium were detected by Western blotting. NF-κB activation was evaluated by electrophoretic mobility shift assay. Results (1) Cardiac function was significantly lower and the level of serum cTnI was significantly higher in the CME group compared with the sham group. CME reduced myocardial systolic dysfunction and left ventricular dilatation. The CME plus nicorandil group showed improved CME-induced cardiac function and reduced serum cTnI level when compared with the CME Group (P < 0.05). (2) Compared with the CME group, the CME plus nicorandil group showed lower PDCD4 and TNF-α expression and NF-κB activity as well as improved cardiac function (P < 0.05). Conclusions The pretreatment of nicorandil effectively reduced the myocardial damage caused by CME, mainly through inhibiting the PDCD4/NF-κB/TNF-α pathway in cardiomyocytes.

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Journal of Medical Postgraduates ; (12): 539-543, 2018.
Article in Chinese | WPRIM | ID: wpr-700869

ABSTRACT

Zika virus (ZIKV), a kind of mosquito borne flavivirus , has spread rapidly in recent years and triggers global large scale epidemic outbreaks.ZIKV infection can lead to severe potential complications such as infant microcephaly and Guillain Barrésyndrome, which has caused social panic and global widespread attentions .The serological detection of ZIKV is easy to be con -fused by other flaviviruses, thus the establishment of specific and effective detection methods is extremely important in the prevention and control of ZIKV transmission.In this paper, the epidemic situation of ZIKV in global and Chinese and research progress of ZIKV detection methods including pathogeny detecting method , serological detecting method and nucleic acid detecting method are reviewed , and the future perspective of detection methods research is also briefly described .

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Article in Chinese | WPRIM | ID: wpr-700541

ABSTRACT

With the development of science and technology,medical simulation has been applied extensively.The application of simulation technique in teaching hospitals has become a new choice in modern medical education.As the most basic and important clinical ability,airway management has become a popular training program in China's teaching hospitals.This paper summarizes the current situation of the simulation training of airway management in teaching hospitals at home and abroad,concludesthe existing problems in the simulation teaching of airway management of teaching hospitals in China,such as narrow range of training objects,lack of professional teaching staff,unscientific curriculum design and outdated training model,etc.Then puts forward the countermeasures of expanding the training coverage,establishing the professional teaching team,designing the curriculum design of science,and introducing advanced simulation equipment to improve the quality of simulated teaching.

18.
Article in Chinese | WPRIM | ID: wpr-697618

ABSTRACT

Objective To investigate the effect of diallyltrisulfide on rats with myocardial injury after coro-nary microembolization (CME). Methods 20 survival of SD rats were randomly divided into CME group (CME group)and diallyltrisulfide pretreatment group(DATSgroup),and these rats were injected with microspheres(42 μm in diameter)into the left ventricles to induce the model of CME, 10 rats for each group.DATS group was received diallyltrisulfide (DATS) 40 mg/(kg·d) for 7 days before operation. Another 10 survival of SD rats was selected as sham operation group(Sham group),and these rats were injected with the same dose of normal saline by left ventri-cles. Cardiac function was assessed by echocardiography and the expression of Akt and caspase-3 of myocardial tissue of rats in each group were detected by TUMEL staining,RT-PCR and Western Blot,while testing the level of cTnI after operation of 6 h. Results Compared with Sham group, the cardiac function of CME group and DATS group was significantly decreased (P<0.05), the expression of caspase3 mRNA and protein was significantly increased (P<0.05), the expression on Akt mRNA and protein was significantly decreased (P<0.05) (P<0.05). Com-pared with CME group, the cardiac function of DATS group was significantly improved (P<0.05), the expression of caspase3 mRNA and protein was significantly decreased (P<0.05), the expression of Akt mRNA and protein was significantly increased (P<0.05), cTnI level was significantly decreased (P<0.05). Conclusion Diallyltrisulfide pretreatment can significantly reduce the apoptosis of cardiomyocytes after CME and improve cardiac function.The mechanism may be through the inhibition of Akt to activate cardiomyocyte caspase3-mediated death receptor activa-tion pathway.

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Article in Chinese | WPRIM | ID: wpr-694377

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Objective To evaluate the prognostic value of transcutaneous oximetry in patients with septic shock.Methods Fifty-three patients with septic shock were enrolled prospectively from January 2013 to December 2015.Transcutaneous oximetry were used to determine the results of 10 min oxygen challenge tests (OCT) carried out at beginning(0 h) and at 6 h after fluid resuscitation respectively.The 10-min OCT value (10 min OCT) and oxygen challenge index(OCI) were calculated.The APACHE Ⅱ and SOFA score,hemodynamic variables,oxygen metabolism indexes,dose of vasoactive agents,10 min OCT,and OCI at 0 h and at 6 h were recorded.Patients were assigned into survival group and death group according to the 28 d survival.The differences in demographics and clinical data were compared between groups.The role of 10 min OCT and OCI in predicting death was evaluated by receiver operating characteristic curves(ROC).The Kaplan-Meier surviving curve was created and the survival of the patients was analyzed by the Log-rank test.Risk factors associated with the prognosis were analyzed using the multiple logistic regression analysis.Results There were 29 patients in the survival group and 24 patients in the death group.Compared with death group,10 min OCT[(77.55±18.48)mmHg vs.(51.30±21.60)mmHg] and OCI [(0.78±0.13) vs.(0.59±0.15)] at 6 h in survival group were significantly higher(P<0.05),while APACHE Ⅱ [(12.48±5.69) vs.(17.25±8.79)] and SOFA [(5.79±1.72) vs.(10.10±2.52)] in survival group were significantly lower than those in death group(P<0.01).The area under the ROC curve of 10 min OCT at 6 h and OCI at 6 h for predicting 28 d death were 0.86±0.05(95%CI:0.76-0.87,P<0.01) and 0.79±0.08(95%CI:0.64-0.95,P<0.01),respectively.The optimal cutoff point for 10 min OCT at 6 h was 72.00 mmHg with the sensitivity of 76.84% and specificity of 85.03%.The optimal cutoff point for OCI at 6 h was 0.76 with the sensitivity of 76.84% and specificity of 77.47%.Kaplan-Meier survival analysis showed that 28 d survival rate in high level of 10 min OCT at 6 h and high level of OCI at 6 h were significantly higher than that in low level of 10 min OCT at 6 h(70.86% vs.31.82%,x2=7.96,P<0.01)and low level of OCI at 6 h (75.00% vs.32.00%,x2=9.86,P<0.01).Multivariate logistic regression analysis showed that both 10 min OCT at 6 h (OR=0.92,95%CI:0.88-0.96,P<0.05) and OCI at 6 h (OR=0.01,95%CI:0.001-0.023,P<0.05) were independent risk factors associated with 28 d mortality of patients with septic shock.Conclusions The 10 min OCT and OCI were reliable predictors for the prognosis of patients with septic shock.

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Article in Chinese | WPRIM | ID: wpr-694359

ABSTRACT

Objective To explore the prognostic value of soluble triggering receptor expressed on myeloid cells-1(sTREM-1) in patients with ventilator-associated pneumonia (VAP).Methods A total of 103 VAP patients were enrolled from June 2013 to May 2015 in the ICU of Wuxi People's Hospital Affiliated to Nanjing Medical University.The demographics and clinical data were collected,while serum sTREM-1,procalcitonin (PCT),C-reactive protein(CRP),clinical pulmonary infection score(CPIS) and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) were measured.Patients were divided into the death group and the survival group according to 28 d survival.The differences in demographics and clinical data were compared between groups.The values of sTREM-1,PCT,CPIS and APACHE Ⅱ for predicting 28 d death were evaluated by receiver operating curves(ROC).The surviving curve was drawn by Kaplan-Meier method.The possible prognostic factors were analyzed by univadate and logistic multivariate analysis.Results There were 76 patients in the survival group and 27 patients in the death group,and there was no difference in demographics between two groups(P>0.05).The serum sTREM-1,PCT,CPIS and APACHE Ⅱ were higher in the death group[(89.50±18.45) pg/mL,(823.86±182.74) pg/ mL,(7.20±1.74) and (19.58±3.43)] than those in the survival group[(54.09±12.71) pg/mL,(579.81±193.45) pg/mL,(4.79±1.93) and (17.23±3.12),all P<0.05].The areas under the ROC of sTREM-1,PCT,CPIS and APACHE Ⅱ for predicting 28 d death were 0.84±0.04(95%CI:0.75-0.92,P<0.01),0.65±0.05(95%CI:0.55-0.74,P=0.49),0.67±0.06(95%CI:0.55-0.79,P<0.01),0.79±0.04(95%CI:0.70-0.87,P=0.03),respectively.Patients were assigned into two groups by the best cutoffpoint of sTREM-l=75.00 pg/mL,and Kaplan-Meier survival analysis showed that 28 d survival rate in the low sTREM-1 group was significantly higher than that in the high sTREM-1 group (82.5% vs.63.4%,x2=3.96,P<0.05).Multivariate logistic regression analysis showed that both sTREM-1 (OR=1.08,95%CI:1.04-1.13,P<0.01) andAPACHE Ⅱ (OR=1.39,95%CI:1.15-1.67,P<0.01) were risk factors associated with 28 d death.Conclusions Early serum sTREM-1 can be used as a reliable predictor for the outcome of patients with VAP.

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