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ObjectiveBody fluid stains left at crime scenes are frequently trace amounts, while the identification of body fluids through real time fluorogenic quantitative technique often necessitates the repeated detection within the limited sample, as multiple miRNA markers are the basis for the identification. Based on the goal of both the throughput and efficiency improvement of miRNA analysis in trace samples, a duplex real time fluorogenic quantitative PCR assay system was designed to accurately quantify two miRNAs simultaneously, and the system should be further verified by actual sample for the body fluid identification. MethodsThe duplex real time fluorogenic quantitative PCR system of miR-451a to miR-21-5p was established with specially designed primers and probes, and the concentrations of the primers and probes were both optimized. The specificity, sensitivity and reproducibility of the system were validated, while its capability for body fluid identification was assessed using the miR-451a to miR-21-5p ratio. ResultsThe optimized assay system exhibited excellent specificity and repeatability, with coefficients of variation consistently below 8% for both intra- and inter-batch variability. The amplification efficiency of miR-451a and miR-21-5p reached 71.77% and 74.81%, respectively, with high and relatively consistent results. By utilizing this duplex real time fluorogenic quantitative PCR assay system, a total of 58 body fluid samples were analyzed, exhibiting a discrimination rate of 100% between blood and non-blood samples, as well as between peripheral blood and menstrual blood samples. Moreover, the results, obtained from single real time fluorogenic quantitative PCR assay system and duplex real time fluorogenic quantitative PCR assay system, showed no statistically significant difference with randomly selected blood samples (n=20). Compared to previous single real time fluorogenic quantitative PCR assay system, the sensitivity of duplex real time fluorogenic quantitative PCR assay system exhibited remarkable improvement. A minimum input of only 0.1 ng total RNA was sufficient for accurate detection of peripheral blood and menstrual blood samples, while saliva, semen, and vaginal secretion required only 1 ng total RNA for precise identification purposes. Additionally, the duplex real time fluorogenic quantitative PCR assay system successfully differentiated between different types of body fluids in simulated samples under natural outdoor conditions. ConclusionThe duplex real time fluorogenic quantitative PCR assay system effectively reduced both the time and material costs by half compared to the single system, especially suitable for the examination of body fluid stains left at crime scenes, solving the contradiction between the trace amount and the multiple sample volumes demand of repeated real time fluorogenic quantitative PCR. The duplex real time fluorogenic quantitative PCR assay successfully distinguished blood and other body fluid, as well as peripheral blood and menstrual blood samples, which maintains an equivalent capability for body fluid identification with half sample, time and reagent consumption. This system provides an efficient tool for identifying suspicious body fluids, as well as a foundation for more multiplexed real time fluorogenic quantitative PCR assay system research.
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Objective:To summarize the clinicopathologic features and clinical diagnosis and treatment experience of retroperitoneal paraganglioma.Methods:This study retrospectively analyzed the clinical, pathological and follow-up data of 39 patients admitted to the First Affiliated Hospital of Xi'an Jiaotong University from 1 Oct 2012 to 1 Oct 2022 for retroperitoneal paragangliomas undergoing resection.Results:There were 19 males and 20 females with tumor being functional in 11 cases (28%) and non-functional in 28 cases (72%). CT angiography showed that the tumors were distributed around the abdominal aorta and inferior vena cava in most cases. All 39 patients underwent tumor rescetion.Patients in laparoscopic group had shorter operation time and postoperative hospital saty compared with open sugery [(135±66)min vs. (194±67)min, t=-2.529, P=0.016; (6.6±2.2)d vs.(9.6±4.8)d, t=-2.096, P=0.043], while there was no statistically significant difference between the two groups in terms of intraoperative blood loss [(152±151)ml vs. (361±608)ml, t=-1.169, P=0.250]. There were no major postoperative complications in the laparoscopic group, and pulmonary infection in 1 case and intestinal obstruction in 1 case in the open group. Thrity-six cases were followed up, ranging from 2 to 115 months, 1 patient in the laparoscopic group died 1 year after surgery due to recurrence and metastasis. In the open group, 1 case recurred 2 years later and was discharged after the second operation, and 1 case died of recurrence 2 years after surgery. Conclusions:Surgery is indicated for retroperitoneal paraganglioma. Adequate perioperative management is the key to the success of the operation. Laparoscopic surgery is superior to open surgery in terms of operation time and postoperative recovery .
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Objective:To explore the clinical characteristics of patients with paraquat mixed with diquat poisoning.Methods:The clinical data of 145 patients with paraquat mixed with diquat poisoning admitted to the Department of Emergency of the First Affiliated Hospital of Wenzhou Medical University from January 20, 2016 to March 31, 2022 were retrospectively analyzed. According to the detection results of plasma toxicants in patients with poisoning, the patients were divided into the paraquat diquat mixed group (mixed group), paraquat group (PQ group) and diquat group (DQ group). The clinical indexes, organ dysfunction, different poisoning doses and prognosis of the three groups were compared. Patients in the mixed group were divided into the survival group and death group according to their 90-day survival, and the differences of each index between the two groups were compared. Kaplan-Meier survival analysis was conducted for each index. After Log-rank test, multivariate Cox regression was used to analyze the risk factors of death in the mixed group.Results:A total of 31 patients were included in the mixed group, 92 patients in the PQ group, and 22 patients in the DQ group. There were significant differences in age, toxic dose, number of organ dysfunction, PSS score and APACHE II score among the three groups ( P<0.05). The main injured organs of the mixed group were gastrointestinal tract, kidney, liver, lung and nervous system. The proportion of organ damage in the mixed group was higher than that in the PQ group and DQ group. The white blood cell count, neutrophil count, HB, creatinine, AST, lactic acid, PT and APTT were statistically significant among the three groups ( P<0.05). In the mixed group, patients taking oral administration of < 20 mL all survived; 8 patients taking oral administration of 20 -50 mL died; 11 patients took oral administration of 51-100 mL and 8 (72.7%) died; and 10 patients took oral administration of more than 100 mL and 9 patients (90%) died. In the mixed group, patients with the concentration of diquat > 5000 ng/mL died. Among 31 patients with mixed poisoning, 30 patients (96.78%) had significantly higher concentrations of diquat than paraquat. There were no significant differences in sex, age, time from poisoning to hospitalization, ingestion amount, lymphocyte count, Hb, BNU, CK, total bilirubin, PH, and PT between the survival group and the death group ( P>0.05). Multivariate Cox regression analysis showed that the ingestion amount, plasma PQ concentration at admission, plasma DQ concentration at admission, and lactic acid were independent risk factors for death ( P<0.05). Conclusions:Paraquat mixed with diquat can cause multiple organ function damage. The main damaged organs are gastrointestinal tract, kidney, liver, lung and nervous system. Compared with PQ or DQ poisoning, mixed poisoning has a higher incidence of organ damage, a more serious condition, and a higher mortality rate. Ingestion amount, plasma PQ concentration at admission, plasma DQ concentration at admission and lactic acid were independent factors influencing the prognosis of mixed poisoning.
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Objective:To explore the risk factors of microvascular invasion (MVI) in hepatocellular carcinoma (HCC), and to predict MVI preoperatively, non-invasively and accurately.Methods:A total of 150 HCC patients (183 HCC lesions) were retrospectively collected in the First Affiliated Hospital of Xi′an Jiaotong University from January 2016 to June 2022.The clinical data and hematological data, gray-scale ultrasonography (US), contrast-enhanced ultrasonography (CEUS), enhanced magnetic resonance imaging with gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (EOB-MRI) and pathological data of these patients were recorded. According to the pathological diagnosis of MVI, the lesions were divided into MVI (+ ) group and MVI (-) group. The indicators between the two groups were compared. All 183 lesions were put into the training set, and the prediction model with nomogram was constructed according to the risk factors of MVI selected by multivariate Logistic regression. The internal verification was carried out by ten-fold cross-validation method.Results:There were significant statistical differences in the following parameters between MVI (+ ) group ( n=109) and MVI (-) group ( n=74) (all P<0.05). These were cirrhosis, serological parameters (alpha-fetoprotein, albumin, total bilirubin), qualitative indexes of US (size, boundary, internal echo), qualitative indexes of CEUS (hyper/iso/hypovascularity of lesions in arterial phase, portal phase, and delayed phase compared with hepatic parenchyma), and quantitative indexes of EOB-MRI [post enhancement rate (post ratio) and gadolinium disodium rate (EOB ratio)] calculated mainly in terms of lesions and surrounding liver parenchyma in hepatobiliary phase and unenhanced T1 images). Finally, cirrhosis of patients, the size, boundary, internal echo of lesions in US; arterial phase (AP), portal phase (PP), post-vascular phase (PVP) features in CEUS; the EOB rate and post rate of EOB-MRI entered the prediction model of MVI. The training set exhibited good calibration and net gain rate. The areas under the ROC curve for the training set and the validation set were 0.981 and 0.961, respectively, while the diagnostic accuracy were 92.9% and 85.8%, respectively. Conclusions:The model constructed mainly by multimodality imaging methods can achieve favorable predictive performance for MVI, which provides valuable ideas for noninvasively predicting the incidence of MVI and optimizing the MVI-related treatment of MVI in HCC patients.
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Objective:To explore the value of vascular endothelial-cadherin (VE-cad) in evaluating the severity of sepsis.Methods:A prospective study was conducted to select 85 patients with sepsis treated in the emergency ward of the First Affiliated Hospital of Wenzhou Medical University from June 1, 2015 to November 1, 2017. The gender, age, medical history, first infection site, number of affected organs, laboratory indexes, acute physiology and chronic health evaluationⅡ(APACHEⅡ), simplified acute physiology score Ⅱ(SAPSⅡ), sequential organ failure assessment (SOFA) and the total length of stay, emergency intensive care unit (EICU) length of stay, 28-day at admission and survival during hospitalization were measured, and the VE-cad level within 24 hours at admission was measured. The patients were divided into sepsis group and septic shock group according to the progress of the disease. The patients were divided into multiple organ dysfunction syndrome (MODS) group and non MODS group according to whether they were accompanied by MODS. The differences of the above indexes in patients with different disease progression, MODS and different prognosis were analyzed and compared. The receiver operator characteristic curve (ROC curve) was drawn to evaluate the value of VE-cad in evaluating the severity of sepsis.Results:A total of 85 patients were included, mainly respiratory tract infection. Among them, 38 cases were sepsis and 47 cases were septic shock, 39 cases had MODS, 46 cases had no MODS, 64 cases survived and 21 cases died within 28 days after admission. Compared with sepsis group, the number of affected organs in septic shock group was greater [3 (2, 4) vs. 1 (0, 2)], APACHE Ⅱscore [13 (10, 21) vs. 7 (5, 12)], SAPS Ⅱscore [35 (31, 55) vs. 7 (5, 12)], SOFA score [7.0 (5.0, 10.0) vs. 3.0 (0, 5.0)], blood lactic acid [Lac (mmol/L): 3.5 (2.4, 6.2) vs. 1.9 (1.2, 2.2)], C-reactive protein [CRP (mg/L): 90.0 (58.1, 90.0) vs. 50.5 (38.0, 90.0)] and VE-cad levels [mg/L: 1.427 (1.141, 2.150) vs. 1.195 (0.901, 1.688)] were significantly increased, while platelet count [PLT (×10 9/L): 113.4±67.2 vs. 202.5±109.5] and hemoglobin (Hb) levels (g/L: 106.3±36.3 vs. 118.6±18.0) were significantly decreased (all P < 0.05). Compared with non MODS group, APACHE Ⅱ score [14 (10, 22) vs. 8 (6, 13)], SAPS Ⅱ score [36 (32, 56) vs. 29 (24, 35)], SOFA score (7.9±3.9 vs. 4.0±3.8), in-hospital mortality [53.8% (21/39) vs. 0% (0/46)], Lac [mmol/L: 3.1 (2.3, 6.3) vs. 2.1 (1.4, 4.6)] and VE-cad levels [mg/L: 1.427 (1.156, 1.937) vs. 1.195 (0.897, 1.776)] in MODS group were significantly higher, the length of stay in EICU was significantly longer [days: 6 (3, 12) vs. 3 (0, 7)], and the PLT level was significantly lower (×10 9/L: 118.2±80.0 vs. 182.5±104.0, all P < 0.05). Compared with the death group, the number of affected organs in the survival group was fewer [2 (1, 3) vs. 3 (1, 5)], APACHE Ⅱ score [9 (6, 13) vs. 21 (13, 25)], SAPS Ⅱ score [31 (25, 36) vs. 55 (35, 63)] and SOFA score (4.7±3.7 vs. 8.9±4.5) were significantly reduced, and the length of stay in EICU [days: 4 (1, 8) vs. 8 (3, 15)] was significantly shorter (all P < 0.05). ROC curve analysis showed that area under the ROC curve (AUC) of VE-cad, SOFA score and VE-cad combined with SOFA score in evaluating the severity of sepsis were 0.632 [95% confidence interval (95% CI) was 0.513-0.750], 0.830 (95% CI was 0.744-0.916) and 0.856 (95% CI was 0.779-0.933), respectively. When the cut-off value of VE-cad was 1.240 mg/L, the sensitivity was 68.1% and the specificity was 55.3%, the sensitivity of VE-cad combined with SOFA score was 85.1%, the specificity was 73.7%. Conclusion:VE-cad has a certain evaluation value for the severity of sepsis, and the evaluation value of VE-cad combined with SOFA score is better than that of VE-cad single index.
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Objective:To analyze the clinical features and prognosis of liver perivascular epithelioid cell neoplasms (PEComa).Methods:The clinical data of 12 patients with liver PEComa diagnosed by pathology at the First Affiliated Hospital of Xi 'an Jiaotong University from October 2011 to November 2021 were retrospectively analyzed. There were 1 male and 11 females, with a median age of 44 (range 20 to 63) years old. The clinical manifestations, laboratory examinations, imaging features, treatment methods, postoperative pathological features and treatment outcomes of these patients were collected and analysed. Postoperative follow-up by telephone or patient's follow-up records.Results:Among the 12 patients with hepatic PEComa, 8 patients (66.7%) were asymptomatic and 11 patients (91.7%) had a single lesion. All patients underwent surgical treatment, including local tumor resection in 10 patients (83.3%) and extended hemihepatectomy in 2 patients (16.7%). Enhanced CT showed the lesion to be a quasi-round homogeneous low-density mass, enhanced in arterial phase with hepatic artery branches in the lesion, and decrease in enhancement degrees in portal vein phase and delayed phase. Postoperative pathology of the lesions in all the 12 patients was benign. Immunohistochemical results showed that the positive rates of melanoma cell markers HMB45, Melan-A and smooth muscle actin were 100.0%(12/12), 83.3%(10/12) and 91.7%(11/12) respectively. The median follow-up period was 27 months, and no recurrence or metastasis was found.Conclusion:Hepatic PEComa occurred commonly in women with obscure symptoms. The lesion was mainly single and it had no correlation with hepatitis. It is easily confused with primary liver cancer and liver metastasis on medical imagings. PEComa expressed markers of both melanocyte and smooth muscle cell, and radical surgical resection resulted in good results.
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Objective To investigate the effects of up-regulation of zinc finger and BTB domain containing 20 (A20) expression on learning and memory and apoptosis of hippocampal neurons in APP
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Objective:To compare the short-term effects of hip muscles strengthening and quadriceps strengthening on knee osteoarthritis (KOA), and discuss the advantages of hip muscles strengthening. Methods:From October, 2015 to May, 2016, 42 old females with KOA were divided into two groups equally. They received hip strengthening (HS group) and quadriceps strengthening (QS group) for two weeks, respectively. The pain, stiffness and physical function scores of Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) were measured. Isokinetic strength peak torque (PT) was assessed for knee extensors and flexors, and 6-minute Walk Test was also evaluated. They were followed up two months later. Results:Before exercise, there was no significant difference in all the indexes between two groups (P > 0.05). Two weeks after exercise, the knee flexor PT (t = -4.038, P = 0.001) and 6-minute walk distance (t = -2.474, P = 0.022) increased in QS group; the pain, stiffness and physical function scores of WOMAC (t > 2.487, P < 0.05), the knee extensor and flexor PT (|t| > 6.370, P < 0.001), and 6-minute walk distance (t = -2.241, P = 0.037) improved in HS group; the scores of WOMAC were lower (t > 2.087, P < 0.05) and the knee extensor PT was higher (t = -5.028, P < 0.001) in HS group than in QS group. At two-months' follow-up, the drop-out rate was significantly lower in HS group than in QS group (χ2 = 13.480, P < 0.001). Conclusion:Hip muscles strengthening is a good choice for KOA in the early stage of treatment, which could avoid the pain in quadriceps training, increase quadriceps strength and improve physical function.
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Objective:To explore the application effect of online-offline mixed teaching mode in the teaching of histology and embryology.Methods:Two classes of students majoring in psychiatry from Batch 2020 were randomized into experimental group ( n = 100), and two classes of students majoring in general practice in 2020 grade were randomized into the control group ( n = 99). The online-offline mixed teaching mode was used in the experimental group, while a traditional teaching mode was used in the control group. The online test scores, final scores and questionnaire satisfaction of the two groups of students were evaluated on the teaching effect. All data were statistically processed by SPSS 19.0, and the comparison between groups was performed by t test. Results:The online test scores of the experimental group [(8.55±1.18) vs. (6.33±0.91)] and final scores [(85.56±3.32) vs. (72.24±2.71)] were significantly higher than those of the control group ( P < 0.05). Compared with the control group, the experimental group's satisfaction scores of the online-offline mixed teaching mode were significantly higher in students' learning interest, learning autonomy, time management ability, teamwork, interactive communication, knowledge understanding and memory ability ( P < 0.05). Conclusion:The online-offline mixed teaching mode can improve the histology and embryology teaching effect and promote the students' interest in learning and autonomous learning ability.
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Purpose@#Systemic inflammatory response is a critical factor that promotes the initiation and metastasis of malignancies including pancreatic cancer (PC). This study was designed to determine and compare the prognostic value of neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), monocyte-to-lymphocyte ratio (MLR), and fibrinogen-to-albumin ratio (FAR) in resectable PC and locally advanced or metastatic PC. @*Materials and Methods@#Three hundred fifty-three patients with resectable PC and 807 patients with locally advan-ced or metastatic PC were recruited in this study. These patients were classified into a training set (n=758) and a validation set (n=402). Kaplan-Meier survival plots and Cox proportional hazards regression models were used to analyze prognosis. @*Results@#Overall survival (OS) was significantly better for patients with resectable PC with low preoperative PLR (p=0.048) and MLR (p=0.027). Low FAR, MLR, NLR (p < 0.001), and PLR (p=0.003) were significantly associated with decreased risk of death for locally advanced or metastatic PC patients. FAR (hazard ratio [HR], 1.522; 95% confidential interval [CI], 1.261 to 1.837; p < 0.001) and MLR (HR, 1.248; 95% CI, 1.017 to 1.532; p=0.034) were independent prognostic factors for locally advanced or metastatic PC. @*Conclusion@#The prognostic roles of FAR, MLR, NLR, and PLR in resectable PC and locally advanced or metastatic PC were different. FAR showed the most prognostic power in locally advanced or metastatic PC. Low FAR was positively correlated with OS in locally advanced or metastatic PC, which could be used to predict the prognosis.
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Objective To investigate the protective effect and mechanism of curcumin on acute lung injury in septic mice.Methods Totally 120 clean BALB/c male mice were randomly (random number) divided into 8 groups:sham group,sepsis group,curcumin control group,curcumin intervention group,negative virus-sepsis group,negative virus-curcumin intervention group,Mfn2 interference-sepsis group,and Mfn2 interference-curcumin intervention group,15 rats in each group.Mice in the sepsis and the curcumin groups were given the cecal ligation and puncture (CLP) mice in the curcumin intervention and curcumin control groups were given curcumin 200 mg/(kg·d) for 1 week,and mice in the negative virus-sepsis group and negative virus-curcumin intervention groups were established by injection of a negative adeno-associated virus in the tail vein.The Mfn2 interference-sepsis and Mfn2 interference-curcumin intervention groups were established by injecting an adeno-associated virus carrying the Mfn2 interference sequence through the tail vein.Mice were sacrificed after 24 h in each group.The degree of lung injury was examined by lung wet-to-dry weight ratio and pathological examination.The inflammatory factors of alveolar lavage fluid including TNF-α and IL-6 were detected by ELISA,the activation of caspase-3,a key molecule for apoptosis,was detected by Western blot,and apoptosis was detected by TUNEL.The data were analyzed by SPSS 22.0 software,the count data was analyzed by x2 test,and the comparison of measurement data between groups was analyzed by one-way ANOVA.Results Compared with the sham group,the wet-to-dry weight ratio of lung tissue in the sepsis group was significantly increased (71.11 ± 3.78 vs 31.11 ± 5.61,P=0.002),the histopathological score was significantly higher (P=0.006),the inflammatory factors TNF-α (P=0.001) and IL-6 (P=0.012) were dramatically increased,and the apoptosis of lung tissue and the expression of caspase-3 cleaved were also significantly increased (P=0.001).Compared with the sepsis group,the wet-to-dry weight ratio and the histopathological score of lung tissue in the curcumin-treated group was significantly lower (32.84 ± 6.15 vs 71.11 ± 3.78,P=0.004),and the inflammatory factors TNF-α(P=0.013) and IL-6 (P=0.003) were obviously decreased,and apoptosis and apoptosis-related protein caspase-3 cleaved expression were also dramatically decreased (P=0.012).After Mfn2 was down-regulated,Mfn2 interference-curcumin intervention group interfered with Mfn2.Compared with the sepsis group,the dry-to-wet weight ratio and the histopathological score of the lung tissue of the mice was not significantly decreased.Further studies found that after down-regulating Mfn2,compared with the Mfn2 interfere-sepsis group,Mfn2 interfere-curcumin intervention group had no such performance.The inflammatory factors TNF-α and IL-6 were not significantly decreased,and the apoptosis of lung tissue and the expression of apoptosis-related protein caspase-3 cleaved were not significantly reduced.Conclusion Curcumin may attenuate acute lung injury in sepsis by up-regulating the expression of Mfn2.
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Objective@#To investigate the effect of cycloartenyl ferulate (CF) on lipopolysaccharide (LPS)-induced apoptosis in human umbilical vein endothelial cells (HUVEC), and to explore its relative mechanism.@*Methods@#Human umbilical vein endothelial cells were cultured in vitro, and the experiment was divided into the normal group, CF group, LPS group, and LPS+CF groups at different concentrations. After the corresponding treatment of cells in each group, the cell viability and apoptosis of each group were tested by the cell counter Kit-8 (CCK-8) assay and TdT-mediated dUTP nick end labeling (TUNEL) assay. The protein expression of Bax, Bcl-2, caspase-3 and the effect of CF on the protein expression of Nrf2/HO-1 pathway were determined by Western blot. One-way ANOVA were performed in multigroup mean comparison, LSD-t test was used to compare the mean values of two samples, and P<0.05 was considered statistically significant.@*Results@#Compared with the LPS group, the survival rate of HUVECs cells in the CF group was significantly increased with different doses, and the survival rate increased significantly in the 320 μmol/L CF group [(69.85±1.2)% vs ( 100.2±1.824)%, P< 0.01]. TUNEL staining showed that compared with the LPS group, the number of apoptotic positive cells in the 320 μmol/L CF group was significantly reduced [(27.33 ± 3.40) vs (11.67 ± 2.04), P<0.01]. Compared with the LPS group, Bcl-2 protein expression level was significantly increased in the CF group at different doses (P<0.01), caspase-3 and Bax protein expression were significantly decreased (P<0.01). Nrf2 protein expression level increased significantly (P<0.01), and HO-1 protein level increased significantly (P<0.01).@*Conclusion@#CF can alleviate LPS-induced apoptosis of HUVEC, which may be related to the increase of bcl-2 expression, the decrease of caspase-3 and Bax protein expression and the activation of Nrf2/ HO-1 signaling pathway.
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Objective To investigate the protective effect of Baicalin on inflammation induced by lipopolysaccharide in H9C2 cardiomyocytes and its possible mechanism. Methods H9C2 myocardial cells were cultured and pretreated with baicalin at the final concentration of 10, 20, 30 μmol/L for 12 hours, then stimulated with LPS at the final concentration of 1 μg/mL for 6 hours. The control group was treated with the same amount of saline to collect cell samples. CCK-8 (The Cell Counting Kit-8) was used to detect cell activity, enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of interleukin-6 (IL-6), interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α), Western blot was used to detect the protein expression levels of NF-κB p65, p-NF-κB p65, p38 MAPK, p-p38 MAPK, IκBα and p-IκBα. SPSS 23.0 statistical software was used. Independent sample t test was used for comparison between two groups, and one-way ANOVA test was used for comparison among multiple groups. Results The survival rate of myocardial cells in the control group was (93.67 +1.453)%. Compared with the control group, the survival rate of H9C2 myocardial cells induced by LPS decreased (P< 0.05). In the control group, the expression of IL-6 in H9C2 myocardial cells was (49.33 +2.42) pg/mL, the expression of TNF-α was (86.33 +1.85) pg/mL, and the expression of IL-1β was (28.67 +4.66) pg/mL. Compared with the control group, the expression levels of IL-6, IL-1β and TNF-α in H9C2 myocardial cells increased after LPS induction (P< 0.05), while the levels of p-NF-κ B p65, p-p38 MAPK and p-I κ B α protein increased (P< 0.05), while the levels of I κ B α protein decreased (P< 0.05), while the expressions of NF-κ B p65 and p38 MAPK protein did not change significantly (P> 0.05). Compared with LPS group, the survival rate of H9C2 myocardial cells in baicalin intervention group increased (P<0.05), the expression levels of IL-6, IL-1β and TNF-a decreased (P < 0.05), the levels of p-NF-κB p65, p-p38 MAPK, p-I κBα protein decreased (P< 0.05), and the level of IκBα protein increased (P< 0.05), while the expression of NF-κB p65 and p38 MAPK did not change significantly. (P>0.05). Conclusions Baicalin may alleviate LPS-induced cardiomyocyte inflammation by inhibiting the activation of NF-kappa B and p38 MAPK, and improve cell survival.
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Objective To investigate the effect of cycloartenyl ferulate (CF) on lipopolysaccharide (LPS)-induced apoptosis in human umbilical vein endothelial cells (HUVEC),and to explore its relative mechanism.Methods Human umbilical vein endothelial cells were culturedin vitro, and the experiment was divided into the normal group, CF group, LPS group, and LPS+CF groups at different concentrations. After the corresponding treatment of cells in each group, the cell viability and apoptosis of each group were tested by the cell counter Kit-8 (CCK-8) assay and TdT-mediated dUTP nick end labeling (TUNEL) assay. The protein expression of Bax, Bcl-2, caspase-3 and the effect of CF on the protein expression of Nrf2/HO-1 pathway were determined by Western blot. One-way ANOVA were performed in multigroup mean comparison, LSD-t test was used to compare the mean values of two samples, andP<0.05 was considered statistically significant.Results Compared with the LPS group, the survival rate of HUVECs cells in the CF group was significantly increased with different doses,and the survival rate increased significantly in the 320 μmol/L CF group [(69.85±1.2)%vs ( 100.2±1.824)%,P< 0.01]. TUNEL staining showed that compared with the LPS group, the number of apoptotic positive cells in the 320 μmol/L CF group was significantly reduced [(27.33 ± 3.40)vs (11.67 ± 2.04),P<0.01]. Compared with the LPS group, Bcl-2 protein expression level was significantly increased in the CF group at different doses (P<0.01), caspase-3 and Bax protein expression were significantly decreased (P<0.01). Nrf2 protein expression level increased significantly (P<0.01), and HO-1 protein level increased significantly (P<0.01).Conclusion CF can alleviate LPS-induced apoptosis of HUVEC, which may be related to the increase of bcl-2 expression, the decrease of caspase-3 and Bax protein expression and the activation of Nrf2/ HO-1 signaling pathway.
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Objective To explore the antioxidant mechanism ofhistone deacetylase 2 (HDAC2) regulating Nrf 2 acetylation in lipopolysaccharide (LPS)-induced type Ⅱ alveolar epithelial cell injury.Methods The experiment was divided into two parts.The first part was the routine culture of type Ⅱ alveolar epithelial cells of mice.The cells were stimulated with different concentrations of LPS (10 ng/ mL,100 ng/mL and 1 000 ng/mL).CCK-8 was used to detect the cell activity at 0 h,6 h,12 h,24 h and 48 h,respectively.The second part:Alveolar epithelial cells of type Ⅱ were cultured and divided into the normal control group (control group),LPS group,HDAC2 lentivirus interference group (siRNA-HDAC2 group) and HDAC2 lentivirus overexpression group (LV-HDAC2 group).The expression of HDAC2 and Nrf2 were detected by Western blot,the acetylation of Nrf2 was detected by immunoprecipitation,and the stability of nrf2 was detected after actinidone action.The activity of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by chemical colorimetry.SPSS 23.0 statistical software was used.LSD-t test was used for comparison between two groups,and one-way ANOVA test was used for comparison among multiple groups.Results Compared with the control group,the expression of HDAC2 protein in the LPS group increased (t=5.974,P=0.027),the acetylation level of Nrf2 decreased (t=7.223,P=0.002),the Nrf2 protein level increased (t=2.929,P=0.043),the protein stability of Nrf2 increased,the SOD activity decreased (t=121,P<0.01),and the MDA content increased (t=10.45,P=0.000 5).Compared with the LPS group,Nrf2 acetylation level decreased in the LV-HDAC2 group (t=1 1.29,P=0.000 4),Nrf2 protein expression increased (t=3.194,P=0.033),Nrf2 protein stability increased,SOD activity increased (t=4.678,P=0.009),and MDA content decreased in the LV-HDAC2 group (t=5.417,P=0.005 6).While the opposite trend was observed in the siRNA-HDAC2 group.Conclusion After LPS stimulation,oxidative stress of type Ⅱ alveolar epithelial cells was aggravated.HDAC2 could decrease the level of Nrf2 acetylation,increase the expression of Nrf2 protein,and alleviate LPS-induced oxidative stress.
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Epimedii Folium is a commonly used traditional Chinese drug, and now still depends on the wild resource. In recent years, with the surge in consumption, the resources are declining, the use of market varieties are constantly changing. In this paper, Production and sales situation of the five species contained in pharmacopoeia(Epimedium brevicornu, E. sagittatum, E. pubescens, E. koreanum and E. wushanense) have been studied on the basis of the existing researches, in-depth investigation and collection of accurate plant specimens and samples. And the origins of Epimedii Folium regulated by the pharmacopoeia have been discussed. At the same time, more exclusive and practical features have been summarized on the basis of observation and comparison with the stereo, optical and stereo-fluorescence microscopy technologies, and refering the related literatures on leaf structure and anatomy.The results of this study will provide a useful reference for the clinical medication, supervision, inspection, and standard drafting and so on.
Sujets)
Médicaments issus de plantes chinoises , Pharmacologie , Epimedium , Chimie , Classification , Plantes médicinales , Chimie , ClassificationRésumé
AIM To establish an HPLC method for the content determination of six constituents in Shiyifang Vinum (Notoginseng Radix et Rhizoma,Dipsaci Radix,Carthami Flos,etc.).METHODS The content determination of notoginsenoside R1,ginsenoside Rg1,ginsenoside Rb1 and asperosaponin Ⅵ was performed on a 30℃ thermostatic Inertsil(R) ODS-3 C18column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1% phosphoric acid flowing at 1.0 mL/min in a gradient manner,and the detection wavelength was set at 203 nm.The content determination of brucine and strychnine was conducted on a 30 ℃ thermostatic Geminni(R) C18 110(A) column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-mixed solution of 0.01 mol/L sodium heptanesulfonate and 0.02 mol/L potassium dihydrogen phosphate flowing at 1.0 mL/min in an isocratic elution manner,and the detection wavelength was set at 260 nm.RESULTS Six constituents showed good linear relationships within their own ranges (r > 0.999 0),whose average recoveries were 98.52%-99.96% with the RSDs of 2.0%-2.3%.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Shiyifang Vinum.
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Objective To investigate the role of miR-10a in CD4+CD25+Treg-mediated immunosuppression during sepsis and its potential role in immunotherapy for sepsis.Methods Sepsis mouse model was established by cecal ligation and puncture(CLP).Balb/c mice of clean grade were sacrificed 1,3,5,and 7 days after operation.Blood as well as spleen samples were harvested at given intervals.The splenic CD4+CD25+Treg cells and CD4+T cells were isolated by MACS microbeads.Cells were cultured,and phenotypes were analyzed by flow cytometry.The miR-10a expressed in Treg cells were detected by Real-time PCR.After administration of LV-mmu-miR-10a-5p-inhibition,the immunosuppressive function have been detected.Statistical analyses were performed using one-way analysis of variance (SPSS 19.0,Chicago,USA) test followed by Dunnett-t test to compare among three or more groups or by Student's t-test to compare between two groups.Results The percentages of splenic Tregs (CD4+CD25+/CD4+T) was (7.34±1.2)% in normal group,and the increase in percentage of Tregs in spleen has been observed in septic mice (P<0.05).The mean fluorescence intensity (MFI) of Foxp3+Treg was increased in septic mice compared with sham group (P<0.05).The expression of miR-10a was significantly elevated on CLP 1-7 day (P<0.05).After down-regulation of miR-10a in septic mice,the percentages of Tregs (CD4+CD25+/CD4+T) was significantly increased in septic mice (P<0.05),the MFI of Foxp3+Treg was increased in septic mice compared with control group (P<0.05).The CD4+T cell proliferative activity in CLP-induced mice was significantly suppressed on CLP 3 day compared with sham group (P<0.05).After down-regulation of miR-10a in septic mice,the CD4+T cell proliferative activity was significantly suppressed compared with control group (P<0.05).Conclusions Treg plays a critical role in immunosuppression in septic mice.Inhibition of miR-10a in vivo could enhence immunesuppression of CD4+CD25+Treg.Therefore miR-10a may participate in the regulation of CD4+CD25+Treg immunosuppression in sepsis and become the target for immunotherapy.
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Objective To evaluate the application of Web problem-based learning (WPBL) mode of respiratory system integration curriculum.Methods Experimentclasses 1 and 2 of the 2014 grades of the Qiqihar Medical University were divided into control group(48 students) and experiment group (48 students).Traditional PBL teaching was conducted in control group:the contents of the case were printed and discussed with the material of each class.WPBL teaching was conducted in experiment group:autonomous learning before class,showing cases by Video,asking questions and discussion in group and reporting results in class.The teaching effect was evaluated by PBL evaluation form,questionnaire and final grade.SSPS 19.0 statistical software was used for data analysis and measurement.T test was conducted among groups.Counting data were tested by Chi-square and the results were expressed as percentage.Results The students in the experiment group were better than those of control groupin the aspects of contents understanding,course interest,clinical thinking ability,classroom atmosphere and the course attitude.The scores of PBL (16.65 ±2.82),experiment exam (8.21 ±0.44) and final grade(76.77 ± 12.31) in the experiment group were higher than those in the control group.There are statistical differences in two groups (P<0.05).Conclusions WPBL teaching can improve students'learning interest,clinical thinking ability and improve the teaching quality of respiratory integration curriculum.
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Objective To discuss the application of blending learning based on MOOC in Histology and Embryology, so as to improve students' learning effectiveness. Methods Nursing undergraduates in class one of 2017 grade were randomly selected as experimental group (n=47) , and nursing undergraduates in class two as control group (n=48), with blending learning and traditional teaching model adopted, respec-tively. SPSS 19.0 was applied to processing the data and T test were used to compare the results of the final exam of this course and the evaluation scale of students' independent learning ability in two groups. Results The experimental scores [(24.22±5.08) vs. (18.49±4.65)], paper test scores [(52.37±6.24) vs. (47.15± 5.99)], and total score [(76.61±7.22) vs. (62.83±7.36)], which shows the scores of experimental group were statistically higher than those of the control group. According to the evaluation scale of students' inde-pendent learning ability, the self-learning ability of the experimental group was better than that of the control group, with significant differences (P<0.05). Conclusion Blending learning can be applied to the teaching of Histology and Embryology, which can effectively improve learning quality, with satisfactory teaching effects to both teachers and students.