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

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

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

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.

BACKGROUND: Capillarization of hepatic sinusoids is an inevitable part in liver fibrosis and cirrhosis, and is a characteristic lesion inducing portal hypertension. However, curcumin effects on the capillarization of hepatic sinusoids and the underlying mechanism remain unclear. OBJECTIVE: To observe the effect of curcumin (a natural polyphenolic compound derived from the rhizome of Curcuma longa)on the microstructure and secretion of hepatic sinusoidal endothelial cells(HSECs),and to further explore its intervention on sinusoidal capillarization and pharmacological action mechanism of anti-liver fibrosis and target sites. METHODS: The rat HSECs were cultured and divided into seven groups: blank control group received no intervention and cells in the other groups were activated by leptin, followed by treatment with nothing (model group), high-, medium- and low-dose of curcumin, colchicine and salvia miltiorrhiza phenolic acid B, respectively, for 48 hours. RESULTS AND CONCLUSION: Under scanning and transmission electron microscopes, with the increasing activation of leptin, the number of fenestrae in HSECs was increased and the aperture was decreased. Curcumin could increase and enlarge narrowed or disappeared fenestrae caused by leptin, attenuated the thickness and scope of extracellular basement membrane, and reduced the degree of capillarization of hepatic sinusoids in a dose-dependent manner. Real-time PCR and ELISA results showed that after activation of leptin, mRNA and protein expression levels of endothelin-1 and vascular endothelial growth factor in HSECs were significantly increased compared with the blank control group (P < 0.05), while the expressions showed a significant decrease after treatment with curcumin in a dose-dependent manner (P < 0.05). There was also a gradient reduction in the protein expression of endothelin-1 and vascular endothelial growth factor in HSECs treated with curcumin. Moreover, all above mRNA and protein expression levels in the high-dose curcumin group were significantly lower than those in the colchicine and salvia miltiorrhiza phenolic acid B groups. In summary, curcumin can significantly alleviate the sinusoidal capillarization, and thus delay the development of liver fibrosis, probably by down-regulating the expression levels of endothelin-1 and vascular endothelial growth factor.

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.

AIM:To study the influence of bone marrow mesenchymol stem cell-drived exosomes(BMSC-exo-somes)on hindlimb activity,and the numbers of reactive astrocytes and residual neurons in spinal cord injury(SCI)rats. METHODS:BMSCs were cultured using the whole bone marrow adherent culture method and surface markers CD 90 and CD34 were verified by flow cytometry.Exosomes were isolated by ultracentrifugation and the morphology of exosomes was observed under transmission electron microscope.The protein markers CD63 and CD9 were verified by Western blot.After exosomes were applied to SCI rats,the Basso,Beattie and Bresnahan locomotor rating scale score,the Nissl staining of the lesion site,and the numbers of reactive astrocytes and residual neurons were assessed at various time points.RESULTS:Transmission electron microscopic observation revealed the presence of saucer -shaped vesicles.BMSC-exosomes were found to express high levels of CD63 and CD9.Compared with injury group,significant improvement of hindlimb activity scores from day 14 after injury in treatment group was observed(P<0.05),and less reactive astrocytes and more residual neu-rons from day 7 after injury were also observed(P<0.05).CONCLUSION:BMSC-exosomes inhibit reactive astrocytes and death of neurons,and improve hindlimb activity in the rats after SCI.

Objective To evaluate the efficacy and safety of the new active-fitation right ventricular lead temporary-permanent pacemaker (TPPM) rersus the traditional temporary transvenous pacing system .Methods Between January 2011 and June 2013, 234 patients had their infected leads removed at our center. A total of 105 (44.9%) patients were pacemaker dependent. Thirty-five patients underwent TPPM implantation and 70 patients had implanted with traditional temporary transvenous pacing system. For traditional temporary pacing, the quadrupole catheter was implanted into the right ventricle through the femoral vein to connect the temporary pacemaker. In TPPM, an active-fixation electrode was implanted into the right ventricular septum through the subclavian and internal jugular veins to connect to the reused permanent pacemaker. parameters from the pacemakers,time for the procedure,the occurance of complications and rates of infection and mortality during the 2 years of follow up were compared between the 2 groups. Results There were more patients with infectious endocarditis in the TPPM group than in the traditional temporary pacing group(22.9% vs. 5.7%,P=0.019). Therefore,the electrode retention time in the TPPM group was longer[2(2,7)d vs.2(2,3)d,P=0.032]and the hospital stay was slightly prolonged[15(14,21)d vs.17(15,25)d,P=0.05]compared with the traditional temporary pacing group.The pacing threshold in the TPPM group was lower than that in the traditional temporary pacing group[(0.7±0.2)V vs.(1.0±0.3)V, P=0.035)].There was no difference in X-ray exposure time between the groups[(24.7±15.4)min vs.(27.5±17.7)min,P=0.242].There were no complications related to bridging in the TPPM group, but 11 patients in the traditional temporary pacing group had developed complications (P=0.009). Conclusions TPPM is effective and safer as compared to traditional temporary pacing for pacemaker-dependent patients with device infection. The operation time does not increase in patients with TPPM implantation.