Adolescent ; Adult ; Aged ; Female ; Humans ; Liver ; physiopathology ; Liver Function Tests ; Male ; Middle Aged ; Severe Acute Respiratory Syndrome ; immunology ; physiopathology ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.

METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.

RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.

CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.

Animals ; Autophagy ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats

OBJECTIVETo investigate the osteoblast-like characteristics of human periodontal ligament cells affected by elastic stress in vitro, and the role of corebinding factor a 1 (cbfa1) in alveolar bone formation during orthodontic tooth movement.

METHODSRat dig-labeled cbfa1 cDNA probe was prepared from SD rat osteoblasts cultured in vitro. Human periodontal ligament cells were cultured on the elastic bottom plate and stimulated by elastic stress using mechanical loading system for cultured cells in vitro. The expression of cbfa1 mRNA was detected by in situ hybridization method.

RESULTSCbfa1 mRNA express in human periodontal ligament cells stimulated by elastic stress and did not express in normal human periodontal ligament cells.

CONCLUSIONIt is suggested that elastic stress plays a role in the differentiation process from human periodontal ligament cells to osteoblast-like cells. Cbfa1 is a transcription factor in alveolar bone remodeling during orthodontic tooth movement.

Adolescent ; Animals ; Cells, Cultured ; Child ; Core Binding Factor alpha Subunits ; DNA-Binding Proteins ; genetics ; Elasticity ; Humans ; Periodontal Ligament ; cytology ; metabolism ; RNA, Messenger ; analysis ; Rats ; Transcription Factors ; genetics

The Notch signaling pathway has been implicated in the regulation of cell-fate decisions such as differentiation of embryo stem cells and neural stem cells into neurons. We cultured human mesenchymal stem cells (hMSCs) in vitro and induced hMSCs to differentiate into neural cells by beta-mercaptoethanol (beta-ME), DMSO and 3-tert-butyl-4-hydroxyanisole (BHA). Immunocytochemistry was utilized to detect neuron-specific enolase (NSE) and Nissl body, and flow cytometry was used to determine cell growth phases. The expressions of signal molecules involved in the Notch pathway such as Notch1, Jagged 1 (JAG1), presenilin 1 (PS1) and hairy and enhancer of split 1(HES1) were observed by RT-PCR and immunofluorescent techniques. The results were as follows: (1) Before induction, the percentage of hMSCs at G(0)/G(1) was 58.5%, and the percentage at S+G(2)/M was 41.5%. After induction, the percentage of hMSCs at G(0)/G(1) increased to 73.1%, 76.2% and 78.1%, respectively on days 2, 4 and 6, and the percentage at S+G(2)/M decreased to 26.8%, 24.8% and 21.9%, respectively; The percentage of NSE-positive cells reached (77+/-0.35) %; Nisslos staining was positive in cytoplasm. (2) Notch1 and JAG1 were both expressed in hMSCs before and after induction, but the mRNA expressions of both Notch1 and JAG1, detected by RT-PCR, decreased obviously after induction(P<0.05). Notch1 mRNA/beta-actin was 1.157, 0.815, 0.756 and 0.570, and JAG1 mRNA/beta-actin was 0.437, 0.350, 0.314 and 0.362, respectively, on days 0, 2, 4 and 6 after induction. The Notch pathway activation participant PS1 mRNA and Notch pathway target gene HES1 mRNA also decreased apparently after induction (P<0.05), and their mRNA/beta-actin was 0.990, 0.449, 0.441, 0.454 and 0.370, 0.256, 0.266, 0.240 on days 0, 2, 4 and 6, respectively. These observations indicate that the expressions of Notch signal molecules were suppressed when hMSCs were induced to differentiate into neural cells. Based on these findings, we propose that low level of Notch signaling activation may contribute to neural cell differentiation.
Basic Helix-Loop-Helix Transcription Factors ; genetics ; Calcium-Binding Proteins ; genetics ; Cell Cycle ; Cell Differentiation ; Flow Cytometry ; Homeodomain Proteins ; genetics ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; Jagged-1 Protein ; Membrane Proteins ; genetics ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Receptor, Notch1 ; genetics ; Receptors, Notch ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Serrate-Jagged Proteins ; Signal Transduction ; Transcription Factor HES-1

Objective:To investigate the role of microRNA (miRNA)-101a in the liver fibrosis induced by activated hepatic stellate cell (HSC), through upregulating IRE1α signaling pathway.Methods:Carbon tetrachloride (CCl 4) induced liver fibrosis model of mice was established. RT-PCR was used to measure the mRNA level of miRNA-101a in liver tissue of mice. Protein level of α smooth muscle actin(αSMA), collagen I and IRE1α were investigated by Western blot. It was divide into Vehicle, TGFβ1, TGFβ1+ miRNA-NC and TGFβ1+ miRNA-M groups. TGFβ1+ miRNA-NC and TGFβ1+ miRNA-M group were transfected with miRNA-101a mimic negative control and miRNA-101a mimic, respectively. After the corresponding treatments, mRNA level of miRNA-101a was detected by RT-PCR, the protein level of αSMA、collagen I and IRE1α were measured by Western blot. Results:Compared to normal mice, the fibrotic deposition in liver tissue of CCl 4 group was increased significantly [(0.17±0.06) vs. (2.09±0.39), P<0.001)]. Protein level of αSMA, collagen I and IRE1α was increased significantly in the model group [(1.00±0.23) vs. (4.09±0.80), (1.00±0.21) vs. (4.98±1.19), (1.00±0.24) vs. (3.27±0.65), all P<0.001)]. While the mRNA level of miRNA-101a was decreased (1.00±0.05) vs. (0.43±0.05), P<0.001). In vitro study, we found that TGFβ1 could inhibit the mRNA expression of miRNA-101a, induced HSC-T6 activation and then up-regulated protein expression of αSMA, collagen I and IRE1α. Compared to TGFβ1+ miRNA-NC group, the expression of miRNA-101a in TGFβ1+ miRNA-M group increased significantly [(0.59±0.19) vs. (1.89±0.20), P<0.001)]. The protein levels of αSMA, collagen I were reduced by over expression of miRNA-101a [(2.65±0.69) vs. (0.84±0.13), (3.15±0.59) vs. (1.31±0.25), all P<0.05)], and the protein content of IRE1α was down-regulated [ (2.63±0.47) vs. (1.03±0.15), P<0.001)]. Conclusion:miRNA-101a may play a critical role in the inhibition of HSC activation and liver fibrogenesis by blocking IRE1α signaling pathway.

Objective:To explore and analyze the reliability and safety of sham feeding in facilitating the recovery of gastrointestinal function after laparoscopic appendectomy (LA), by using a new device, the Artificial Intelligence Bowel Tone Monitoring System.Methods:The data of 100 cases in Shaanxi Provincial People′s Hospital from Dec. 2020 to Sep. 2022 with acute appendicitis operated by LA who met the inclusion criteria. In this prospective study, participants were divided by random number table into a control group and an experimental group, with 50 cases in each group. The control group performed routine postoperative LA care, and the experimental group performed routine postoperative LA care and sham-feeding state care. The age, gender, recovery time of postoperative bowel sounds, time of first postoperative anal discharge, postoperative nausea and vomiting, abdominal distention, dry mouth and halitosis, and postoperative abdominal pain and other complications were recorded. GraphPad Prism 9.0 and SPSS 22.0 software were adopted to conduct data organization and analysis.Results:There were 100 valid cases in this trial. There were no statistical differences between the two groups in terms of gender, age, duration of surgery, abdominal pain and other symptoms ( P>0.05). The recovery time of bowel sounds after surgery was (8.92±0.56) h in the experimental group and (10.55±0.88) h in the control group, which was statistically significant ( t=10.99, P<0.0001); the recovery time of bowel sounds after surgery was (20.10±0.50) h in the experimental group and (20.96±0.59) h in the control group. There was a statistically significant difference between the two groups ( t=7.84, P<0.0001); there was a statistically significant difference between the experimental group (22%) and the control group (42%) for postoperative nausea and vomiting ( χ2=4.60, P=0.032); there was a statistically significant difference between the experimental group (16%) and the control group (52%) for postoperative abdominal distension ( χ2= There was a statistical difference between the experimental group (40%) and the control group (68%) ( χ2=7.89, P=0.005). The number of hospitalization days in the control group was (11.40±2.47) days and the days in the experimental group was (9.30±2.01) d, the difference between the two groups was statistically significant ( t=4.65, P<0.001); the hospitalization cost in the control group was (27 270.11±2 645.30) yuan and the cost in the experimental group was (23 669.68±2 841.28) yuan, the difference between the two groups was statistically significant ( t=6.56, P<0.001). Conclusion:To a certain extent, sham feeding can accelerate the recovery of gastrointestinal function in patients after LA, reduce the common postoperative discomfort, length of stay and hospital costs of patients.

OBJECTIVETo construct inducible lentiviral vector containing human Notch1 intracellular domain (NICD) gene and enhanced green fluorescent protein (EGFP), and to study its expression in PC12 cells.

METHODSNICD cDNA was amplified by RT-PCR from human placenta tissue. EGFP gene was amplified by PCR from pEGFP-C1. Both NICD and EGFP were cloned into pcDNA 3.1 (+) plasmid to form pcDNA3.1-Notch1-EGFP. Then the Notch1-EGFP fragment was separated and cloned into pLVX-Tight-puro to form pLVX-Notch1-EGFP. The lentivirus were packaged and harvested, which were used to infect PC12 cells. After antibody selection for 2 weeks, the PC12 cells were induced by doxycycline (Dox). The expression of Notch1-EGFP was detected by fluorescence microscope and flow cytometry.

RESULTSThe recombinant inducible lentiviral vectors (pLVX-Notch1-EGFP) were success fully constructed. The EGFP positive cell percentage was over 90% in transfected PC12 cells after 500 ng/ml Dox induction for 36 h. The expression of Notch1 was posited correlated to the Dox concentration. The expression of Notch1 increased with the duration of Dox induction, which got the peak at 36 h after Dox induction.

CONCLUSIONThe recombinant inducible lentiviral vectors containing Notch1 and EGFP gene are successfully constructed, which provides an effective and simple method to regulate the expression of Notch1 in PC12 cells.

Animals ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Lentivirus ; genetics ; PC12 Cells ; Plasmids ; Rats ; Receptor, Notch1 ; genetics ; Transfection

Sinisan is a widely used traditional Chinese medicine (TCM) in treating various diseases; however, the in vivo metabolic profile of its multiple components remains unknown. In this paper, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was applied to identify the metabolites of Sinisan extract in rat plasma, urine, feces and bile after intragastric administration. Using MS(E) and mass defect filter techniques, 41 metabolites of 10 parent compounds (naringin, naringenin, hesperidin, neohesperidin, liquiritin, liquiritigenin, glycyrrhizic acid, glycyrrhetinic acid, saikosaponin a and saikosaponin d) were detected and tentatively identified. It was shown by our results that these compounds was metabolized to the forms of hydroxylation, glucuronidation, sulfation, glucuronidation with sulfation and glucuronidation with hydroxylation in vivo.
Administration, Oral ; Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; metabolism ; pharmacokinetics ; Flavanones ; analysis ; metabolism ; pharmacokinetics ; Glucosides ; analysis ; metabolism ; pharmacokinetics ; Glycyrrhizic Acid ; analysis ; metabolism ; pharmacokinetics ; Hesperidin ; analogs & derivatives ; analysis ; metabolism ; pharmacokinetics ; Hydroxylation ; Male ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Objective:To explore the value of echocardiography for diagnosing infectious endocarditis (IE).Methods:A total of 487 patients with cardiovascular implantable electronic devices (CIED) infection treated in our hospital from 2013-01 to 2015-06 were enrolled.Based on symptoms,blood culture and echocardiography,9 patients with suspected IE were further examined by 18F-FDG PET-CT to confirm their diagnosis and classification.Definitive therapy was conducted and the patients were followed-up for 1 year to confirm the diagnostic accuracy of echocardiography on CIED induced IE.Results:3 patients were preliminarily diagnosed for bacteremia since no vegetation was found by echocardiography,while IE was finally diagnosed by PET-CT.2 patients were preliminarily diagnosed for IE by echocardiography presented valvular vegetation,while PET-CT showed no evidence of vegetation;then one of them was diagnosed as bacteremia by positive blood culture and another was diagnosed as non-infection.4 patients were preliminarily diagnosed for IE by echocardiography indicated existing vegetation after CIED lead extraction,while PET-CT demonstrated no infection sign in heart chamber and the finally diagnosed was as "non-infectious fibrous residual tissue".According to final diagnosis,definitive therapies were performed to specific patients with at least 1 year follow-up study,no one had new and recurrent infection.Conclusion:Echocardiography had deficiency for diagnosing vegetation in heart chamber especially in suspicious IE patients after CIED lead extraction.It is necessary to make accurate diagnosis with other method for guiding appropriate therapy.

Objective: To explore the prevalence and risk factors of orthostatic hypotension (OH) in elderly hypertension patients. Methods: A total of 532 retired hypertension patients elder than 65 years in Guangzhou military region were enrolled. The patients were divided into 2 groups: Hypertension group, n=414 and Hypertension combining OH (H+OH) group, n=118. The patient's age (65-79、≥ 80), hypertension grade (Grade 1-3) and complication status were studied. The risk factors for H+OH prevalence were analyzed by multivariate Logistic regression analysis. Results: The incidence rate of H+OH was 22.2％ (118/532). In H+OH group, the ratios of elderly and very elderly patients were 6.7％ and 23.1％, P<0.05 and the ratios of OH occurrence for hypertension grade 1, 2 and 3 were 12.6％, 23.3％ and 25.2％ respectively, P<0.05. Multivariate Logistic regression analysis presented that systolic blood pressure (BP) in supine position, BP at immediate standing, heart rate in supine position, heart rate after 2 minutes standing and chronic cardiac insufficiency were the impact factors for H+OH occurrence, P<0.05. Conclusion: In elderly hypertension patients, incidence of OH was increasing with age elevating; H+OH has been related to age, severity of hypertension and chronic cardiac insufficiency.