Objective To observe the effect of m⁶A methylation regulation on Notch1 pathway on the homing of BMSCs in asthma, and the intervention study of traditional Chinese medicine compound Yanghe Pingchuan Granules. Methods Rat bone mesenchymal stem cells(BMSC)and bronchial epithelial cells were cocultured. The extracted cells were divided into: bronchial epithelial cell group, asthma bronchial epithelial cell+mesenchymal stem cell co-culture group (co-culture group), co-culture cell+normal serum group, coculture cell+serum containing optimal drug group, siRNA FTO+normal serum group, siRNA FTO-NC+normal serum group, and siRNA FTO+serum containing optimal drug group. The vitality and cell cycle changes of co-cultured cells were detected. The level and markers of homing BMSC were detected by immunofluorescence staining. The expression of Notch1 pathway related genes were detected by qRT-PCR. The expression of Notch1 pathway related proteins were detected by Western blot. Results Compared with bronchial epithelial cell group, the co-cultured cell group showed an increase in the homing level of BMSCs and the expression of C-X-C motif chemokine receptor 4 (CXCR4), stromal cell-derived factor 1 (SDF-1), Notch1, transcription factor recombination signal binding protein-J (RBP-J), and hairy enhancer of split 1 (Hes1) proteins. Compared with the co-cultured cell group and co-cultured cell+normal serum group, the co-cultured cell+serum containing optimal drug group showed an increase in the homing level of BMSCs and the expressions of CXCR4 and SDF-1, while the protein and mRNA levels of Notch1 and Hes1 decreased. Compared with the siRNA FTO-NC+normal serum group, the siRNA FTO+normal serum group showed an increase in the levels of Notch1, activated Notch1, RBP-J, Hes1 protein, and cell viability, while the level of homing BMSC decreased. Compared with siRNA FTO+normal serum group, the levels of Notch1, RBP-J mRNA, activated Notch1, and Hes1 protein decreased, while the level of homing BMSCs increased in siRNA FTO+serum containing optimal drug group. The levels of Notch1, RBP-J, and Hes1 mRNA were reduced in the co-cultured cells+serum containing optimal drug group. Compared with siRNA FTO+serum containing optimal drug group, the expressions of Notch1, activated Notch1, RBP-J, Hes1 protein and cell viability decreased, while the level of homing BMSCs increased in the co-cultured cells+serum containing optimal drug group. Conclusion Yanghe Pingchuan Granules may promote the homing of BMSCs in asthma and alleviate asthma inflammation by upregulating the expression of FTO and inhibiting the expression of downstream genes in the Notch1 signaling pathway.
Animals ; Receptor, Notch1/genetics* ; Mesenchymal Stem Cells/cytology* ; Asthma/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Signal Transduction/drug effects* ; Rats ; Coculture Techniques ; Alpha-Ketoglutarate-Dependent Dioxygenase FTO/genetics* ; Epithelial Cells/metabolism* ; Rats, Sprague-Dawley ; Cells, Cultured ; Male

OBJECTIVES: To investigate the mechanism mediating the regulatory effect of miR-155-5p on proliferation of human submandibular gland epithelial cells (HSGECs) in primary Sjogren's syndrome (pSS). METHODS: Dual luciferase reporter assay was used to verify the targeting relationship between miR-155-5p and the PI3K/AKT pathway. In a HSGEC model of pSS induced by simulation with TRAIL and INF-γ, the effects of miR-155-inhibitor-NC or miR-155 inhibitor on cell viability, cell cycle, apoptosis and proliferation were evaluated using CKK8 assay, flow cytometry and colony formation assay. ELISA and RT-PCR were used to detect the expressions of inflammatory cytokines and miR-155-5p mRNA in the cells; Western blotting was performed to detect the expressions of proteins in the PI3K/AKT signaling pathway. RESULTS: Dual luciferase assay showed that miR-155-5p targets the PI3K/AKT pathway via PIK3R1 mRNA. The HSGEC model of pSS showed significantly decreased cell viability, cell clone formation ability and expressions IL-10 and IL-4 and increased cell apoptosis, cell percentage in G2 phase, expressions of TNF‑α, IL-6, miR-155-5p and PIK3R1 mRNA, p-PI3K/PI3K ratio, p-Akt/AKT ratio, and PIK3R1 protein expression. Treatment of the cell models with miR-155 inhibitor significantly increased the cell viability, G1 phase cell percentage, colony formation ability, and expressions of IL-10 and IL-4 levels, and obviously reduced cell apoptosis rate, G2 phase cell percentage, expressions of TNF-α, IL-6, miR-155-5p and PIK3R1 mRNA, p-PI3K/PI3K ratio, p-AKT/AKT ratio, and PIK3R1 protein expression. CONCLUSIONS In HSGEC model of pSS, inhibition of miR-155-5p can promote cell proliferation and reduced cell apoptosis by targeting PI3K1 mRNA to negatively regulate the overexpression of PI3K/AKT signaling pathway.
Humans ; MicroRNAs/genetics* ; Cell Proliferation ; Signal Transduction ; Proto-Oncogene Proteins c-akt/metabolism* ; Sjogren's Syndrome/pathology* ; Epithelial Cells/cytology* ; Submandibular Gland/cytology* ; Phosphatidylinositol 3-Kinases/metabolism* ; Apoptosis ; Class Ia Phosphatidylinositol 3-Kinase ; Cells, Cultured

Objective:To evaluate the effect of buccal acupuncture on analgesia after tonsilloadenoidectomy in pediatric patients.Methods:This was a randomized controlled study. One hundred and twenty-six American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ pediatric patients, aged 3-12 yr, weighing 12-34 kg, with body mass index <30 kg/m 2, undergoing elective tonsilloadenoidectomy with general anesthesia, were divided into 2 groups ( n=63 each) by the random number table method: buccal acupuncture group (group B) and control group (group C). All pediatric patients received the same anesthesia induction and intraoperative anesthesia maintenance. The concentration of sevoflurane was adjusted to keep the fluctuation amplitude of vital sign parameters within 20% of the baseline value. After surgery, the drug was immediately stopped and the children were transferred to the postanesthesia care unit for resuscitation under general anesthesia. In group B, the bilateral neck points, upper neck points, hologram points on the head and Zhongjiao points were selected before removal of the tracheal catheter, and disposable acupuncture needles were inserted directly into the acupoints and remained for 20-30 min. Group C received no buccal acupuncture. The pain Assessment Scale (FLACC) was used to assess the severity of postoperative pain. The postoperative agitation score was evaluated by Aono four-point rating method to evaluate the occurrence of agitation. The effective pressing times of patient-controlled analgesia, rescue analgesia and occurrence of nausea and vomiting within 48 h after operation were recorded. The occurrence of bleeding, infection and broken needle at acupuncture sites was recorded. Results:Compared with group C, the effective pressing times of patient-controlled analgesia and incidence of nausea and vomiting were significantly decreased in group B ( P<0.05). There was no significant difference in the rate of rescue analgesia and incidence of postoperative agitation between the two groups ( P>0.05). No infection or broken needle was found at acupuncture sites after buccal acupuncture, only 2 cases had slight bleeding at the puncture site, and there was no abnormality after pressing in group B. Conclusions:Buccal acupuncture can enhance the analgesic effect after tonsilloadenoidectomy in pediatric patients.

OBJECTIVES: To observe the role of miR-139-5p and Notch1 signaling pathway in regulation of homing of bone mesenchymal stem cells (BMSCs) of asthmatic rats. METHODS: Normal rat BMSCs were co-cultured with bronchial epithelial cells from normal or asthmatic rats, followed by transfection with miR-139-5p mimics or a negative control sequence. The changes in cell viability and cell cycle were analyzed, and the cellular expressions of CXCR4 and SDF-1 were detected using immunofluorescence staining. The changes of BMSC homing after the transfection were observed, and the expressions of Notch1, RBP-J, and Hes1 mRNAs and proteins and Th1/Th2 cytokines were detected with RT-qPCR, Western blotting or ELISA. RESULTS: The co-cultures of BMSCs and asthmatic bronchial epithelial cells showed significantly decreased expressions of miR-139-5p, IL-2 and IL-12 and increased expressions of CXCR4, SDF-1, IL-5, IL-9, Notch1, RBP-J, and Hes1. Transfection with miR-139-5p mimics significantly increased the expressions of miR-139-5p, IL-2, CXCR4 and SDF-1 and lowered the expression levels of IL-5, IL-9, Notch1, activated Notch1, and Hes1 in the co-cultured cells. Correlation analysis showed that BMSC homing was positively correlated with miR-139-5p and IL-12 and negatively correlated with IL-5 expression. The expression of CXCR4 was negatively correlated with activated Notch1, and SDF-1 was positively correlated with miR-139-5p but negatively correlated with Notch1 expression. CONCLUSIONS High expression of miR-139-5p promotes homing of BMSCs in asthma by targeting the Notch1 signaling pathway to regulate the expressions of Th1/Th2 cytokines, thereby alleviating airway inflammation.
Asthma/genetics* ; Animals ; Mesenchymal Stem Cells/cytology* ; MicroRNAs/metabolism* ; Rats ; Transcription Factor HES-1/genetics* ; Signal Transduction ; Receptor, Notch1/genetics* ; Immunoglobulin J Recombination Signal Sequence-Binding Protein/genetics* ; Receptors, CXCR4/genetics* ; Coculture Techniques ; Rats, Sprague-Dawley ; Chemokine CXCL12/genetics* ; Epithelial Cells/metabolism*

Objective:To investigate the safety and feasibility of the CHESS endoscpic ruler (CHESS ruler), and the consistency between the measured values and the interpretation values by endoscopic physician experience.Methods:From January 2021 to January 2022, a total of 105 liver cirrhosis patients with portal hypertension were prospectively enrolled from General Hospital, Xixia Branch Hospital, Ningnan Hospital of People′s Hospital of Ningxia Hui Autonomous Region (29 cases), and the First People′s Hospital of Yinchuan (25 cases), General Hospital of Ningxia Medical University (18 cases), Wuzhong People′s Hospital (10 cases), the Fifth People′s Hospital of Ningxia Hui Autonomous Region (10 cases), Shizuishan Second People′s Hospital (6 cases), Yinchuan Second People′s Hospital (5 cases), and Zhongwei People′s Hospital (2 cases) 8 hospitals. The clinical characteristics of all the patients, including gender, age, nationality, etiolog of liver cirrhosis, and Child-Pugh classification of liver function were recorded. A big gastroesophageal varices was defined as diameter of varices ≥5 mm. Endoscopist (associated chief physician) performed gastroscopy according to the routine gastroscopy procedures, and the diameter of the biggest esophageal varices was measured by experience and images were collected, and then objective measurement was with the CHESS ruler and images were collected. The diameter of esophageal varices of 10 randomly selected patients (random number table method) was determined by 6 endoscopists (attending physician or associated chief physician) with experience or measured by CHESS ruler. Kappa test was used to test the consistency in the diameter of esophageal varices between measured values by CHESS ruler and the interpretation values by endoscopic physician experience.Results:Among 105 liver cirrhosis patients with portal hypertension, male 65 cases and female 40 cases, aged (54.8±12.2) years old, Han nationality 82 cases, Hui nationality 21 cases and Mongolian nationality 2 cases. The etiology of liver cirrhosis included chronic hepatitis B (79 cases), alcoholic liver disease (7 cases), autoimmune hepatitis (7 cases), chronic hepatitis C (2 cases), and other etiology (10 cases). Liver function of 32 cases was Child-Pugh A, Child-Pugh B 57 cases, and Child-Pugh C 16 cases. All 105 liver cirrhosis patients with cirrhotic portal hypertension were successfully measured the diameter of gastroesophageal varices by CHESS ruler, and the success rate of application of CHESS ruler was 100.0% (105/105). The procedure time from the CHESS ruler into the body to the exit of the body after measurement was (3.50±2.55) min. No complications happened in all the patients during measurement. Among 105 liver cirrhosis patients with cirrhotic portal hypertension, 96 cases (91.4%) were recognized as big gastroesophageal varices by the endoscopists. Totally 93 cases (88.6%) were considered as big gastroesophageal varices by CHESS ruler. Eight cases were recognized as big gastroesophageal varices by the endoscopist, however not by the CHESS ruler; 5 cases were recognized as big gastroesophageal varices by the CHESS ruler, but not by the endoscopists; 4 cases were not recognized as big gastroesophageal varices both by the endoscopists and CHESS ruler; 88 cases were recognized as big gastroesophageal varices both by the endoscopists and CHESS ruler. The missed diagnostic rate of big gastroesophageal varices by the endoscopists experience was 5.4% (5/93), and the Kappa value of consistency coefficient between the measurement by the CHESS ruler and the interpretation by endoscopists experience was 0.31 (95% confidence interval 0.03 to 0.60). The overall Kappa value of consistency coefficient by 6 endoscopists measured by CHESS ruler in big gastroesophageal varices diagnosis was 0.77 (95% confidence interval 0.61 to 0.93).Conclusion:As an objective measurement tool, CHESS ruler can make up for the deficiency of subjective judgment by endoscopists, accurately measure the diameter of gastroesophageal varices, and is highly feasible and safe.

Based on the surgical model using transforaminal lumbar interbody fusion (TLIF) to treat lumbar spondylolisthesis, this paper presents the investigations of the biomechanical characteristics of cage and pedicle screw in lumbar spinal fusion implant fixed system under different combinations with finite element method. Firstly, combining the CT images with finite element pretreatment software, we established three dimensional nonlinear finite element model of human lumbar L4-L5 segmental slight slippage and implant under different fixed combinations. We then made a comparison analysis between the biomechanical characteristics of lumbar motion range, stress distribution of cage and pedicle screw under six status of each model which were flexion, extension, left lateral bending, right lateral bending, left axial rotation and right axial rotation. The results showed that the motion ranges of this model under different operations were reduced above 84% compared with those of the intact model, and the stability of the former was improved significantly. The stress values of cage and pedicle screw were relatively larger when they were fixed by single fusion device additional unilateral pedicle screw, but there was no statistically significant difference. The above research results would provide reference and confirmation for further biomechanics research of TLIF extracorporal specimens, and finally provide biomechanical basis for the feasibility of unilateral internal fixed diagonal intervertebral fusion TLIF surgery.
Biomechanical Phenomena ; Finite Element Analysis ; Humans ; Lumbar Vertebrae ; Models, Anatomic ; Motion ; Pedicle Screws ; Posture ; Range of Motion, Articular ; Rotation ; Spinal Fusion

In the present study, a finite element model of L4-5 lumbar motion segment was established based on the CT images and a combination with image processing software, and the analysis of lumbar biomechanical characteristics was conducted on the proposed model according to different cases of flexion, extension, lateral bending and axial rotation. Firstly, the CT images of lumbar segment L4 to L5 from a healthy volunteer were selected for a three dimensional model establishment which was consisted of cortical bone, cancellous bone, posterior structure, annulus, nucleus pulposus, cartilage endplate, ligament and facet joint. The biomechanical analysis was then conducted according to different cases of flexion, extension, lateral bending and axial rotation. The results showed that the established finite element model of L4-5 lumbar segment was realistic and effective. The axial displacement of the proposed model was 0.23, 0.47, 0.76 and 1.02 mm, respectively under the pressure of 500, 1 000, 1 500 and 2 000 N, which was similar to the previous studies in vitro experiments and finite element analysis of other people under the same condition. The stress distribution of the lumbar spine and intervertebral disc accorded with the biomechanical properties of the lumbar spine under various conditions. The established finite element model has been proved to be effective in simulating the biomechanical properties of lumbar spine, and therefore laid a good foundation for the research of the implants of biomechanical properties of lumbar spine.
Biomechanical Phenomena ; Finite Element Analysis ; Humans ; Intervertebral Disc ; anatomy & histology ; Lumbar Vertebrae ; anatomy & histology ; Models, Anatomic ; Pressure ; Prostheses and Implants ; Range of Motion, Articular ; Rotation

Objective Colorectal polyp is a precancerous lesion of colorectal cancer .Aim of the study was to explore the risk fac-tors of colorectal polyp malignant transformation .Methods The related information of 75 084 colonoscopies performed from 2003 to 2012 in Southwest Hospital in Chongqing were collected and the relationship between polyp malignant transformation and the pa-tient age ,sex ,polyp location ,size or histological types was analyzed .Results From 2003 to 2012 ,polyps were diagnosed in 14 806 cases of the total 75 084 patients with a 19 .72% detection rate .There were significant difference of the left-side and right-side pol-yp detection rate in different age groups ,and the frequency of polyps distributed in the whole colorectum increased with the increase of age .The rates of epithelial neoplasia and malignant transformation increased with age .At the same time ,malignant transforma-tion rate was significant higher in polyps located in left-side than that in right-side (P<0 .0167) ,in adenoma than that in inflamma-tory hyperplastic polyp (P<0 .01) .The larger diameter and the more villus ,the higher rate of malignant transformation .Conclusion Patient age ,polyp size ,location and histological type could be considered as the significant predictors of colorectal polyp malignant transformation .It may be useful to treat the polyp with endoscopy in patient with age more than 45 and adenoma whose diameter was not less than 1 cm ,located in left-side for prevention of colorectal cancer .