Objective:To investigate the mechanism of proanthocyanidin (PA) in regulating the osteogenic differentiation of human periodontal ligament stem cells (PDLSCs), and to explore the effects of PA on the expression and nuclear translocation of transcription factor EB (TFEB) and on the autophagy-lysosome pathway.Methods:PDLSCs were divided into control group and PA group, which were subjected to RNA sequencing analysis (RNA Seq) to detect differentially expressed genes. The osteogenic differentiation ability and autophagy level were observed by real-time fluorescence quantitative PCR (RT-qPCR) analysis, alkaline phosphatase (ALP) staining and transmission electron microscope (TEM), respectively. Scratch assay and Transwell assay were used to detect the migration ability of PDLSCs. Lysotracker and immunofluorescence staining were used to detect the biogenesis of lysosomes. The total protein expression of transcription factor EB (TFEB) as well as that in cytoplasm and nucleus were detected by Western blotting. Confocal laser scanning microscope (CLSM) was used to observe the nuclear translocation of TFEB. The PDLSCs were treated with small interfering RNA (siRNA) technology to knock down the expression levels of TFEB gene with or without PA treatment. Western blotting was used to analyze the expressions of autophagy-related proteins Beclin1 and microtubule-associated protein 1 light chain 3 (LC3B), as well as osteogenic-related proteins runt-related transcription factor 2 (RUNX2), ALP, and osteocalcin in PDLSCs.Results:Compared with the control group, the osteogenic-related and autophagy-related genes showed differential expression in PDLSCs after PA treatment ( P<0.05). The mRNA expression levels of osteogenic-related genes RUNX2 (2.32±0.15) and collagen type Ⅰ alpha 1 (COL1α1) (1.80±0.18), as well as the autophagy related genes LC3B (1.87±0.08) and Beclin1 (1.63±0.08) were significantly increased in the PA group, compared with the control group (1.01±0.16, 1.00±0.10, 1.00±0.07, 1.00±0.06, respectively, all P<0.01). Compared with the control group, the PA group had higher ALP activity, and more autophagosomes and autophagolysosomes observed by TEM. PA promoted the migration of PDLSCs ( P<0.05) and the increased number of lysosomes and the expression of lysosomal associated membrane protein 1 (LAMP1). In the PA group, the relative expression level of total TFEB protein (1.49±0.07) and the nuclear/cytoplasmic expression of TFEB protein (1.52±0.12) were significantly higher than the control group (1.00±0.11, 1.00±0.13, respectively) ( t=6.43, P<0.01; t=5.07, P<0.01). The relative nuclear/cytoplasmic fluorescence intensity of TFEB in the PA group (0.79±0.09) was increased compared with the control group (0.11±0.08) ( t=8.32, P<0.01). Knocking down TFEB significantly reduced the expression of TFEB (1.00±0.15 vs 0.64±0.04), LAMP1 (1.00±0.10 vs 0.69±0.09), Beclin1 (1.00±0.05 vs 0.60±0.05), and LC3B Ⅱ/Ⅰ (1.00±0.06 vs 0.73±0.07) in PDLSCs ( P<0.05, P<0.05, P<0.01, P<0.01). When TFEB gene was knocked down, the expression levels of Beclin1 (1.05±0.11), LC3B Ⅱ/Ⅰ (1.02±0.09), RUNX2 (1.04±0.10), ALP (1.04±0.16), and osteocalcin (1.03±0.15) proteins were significantly decreased in the PA group compared with the pre-knockdown period (1.28±0.03, 1.44±0.11, 1.38±0.11, 1.62±0.11, 1.65±0.17, respectively) ( P<0.05, P<0.01, P<0.05, P<0.01, and P<0.01, respectively). Conclusions:PA promotes the osteogenic differentiation of PDLSCs through inducing the expression and nuclear translocation of TFEB and activating the autophagy-lysosome pathway.

Objective:To analyze the differences in clinical and endoscopic ultrasonography (EUS) findings between diffuse and focal IgG4-related autoimmune pancreatitis (IgG4-AIP).Methods:Data of patients diagnosed as having IgG4-AIP who underwent EUS at Chinese PLA General Hospital from September 2011 to April 2022 were retrospectively collected. General clinical data, EUS features, and postoperative pathology were analyzed for characteristic differences.Results:A total of 40 patients were included in the study, 60.03±10.87 years old, a higher proportion of males (85.0%, 34/40). All patients underwent EUS, and 28 underwent EUS-guided fine-needle aspiration. Among the 40 patients, 29 (72.5%) had diffuse type and 11 (27.5%) had focal type. Abdominal pain [65.5% (19/29) VS 18.2% (2/11), χ2=5.393, P=0.020] and thickening of the bile duct wall [51.7% (15/29) VS 9.1% (1/11), χ2=4.394, P=0.036] were more common in the diffuse type, while main pancreatic duct dilation [45.5% (5/11) VS 10.3% (3/29), χ2=4.146, P=0.042] was more common in the focal type, with the lesion most commonly located in the pancreatic head (90.9%, 10/11). There was no significant difference in the presence of chronic pancreatitis parenchymal changes between the two groups [34.5% (10/29) VS 27.3% (3/11), χ2=0.003, P=0.955]. Conclusion:There are certain differences in abdominal pain and biliary and pancreatic duct lesions between diffuse and focal AIP. The high expression of chronic pancreatitis characteristics is not observed in either group, which provides clues for the classification of AIP in clinical practice.

The molar anchorage control in orthodontic treatment is a key concern of clinicians and a hot spot in the field of orthodontic clinical research.Good molar anchorage control is a prerequisite for the success of orthodontic treatment.In recent years,clear aligner treatment has been favored by orthodontists and patients because of its aesthetics,comfort and other advantages.However,the unique biomechanical mechanism of clear aligner system has brought new changes and challenges for dentists to understand the anchorage con-trol in orthodontics.This article provides a systematic review of the research methodology,clinical efficacy and enhanced strategy of mo-lar anchorage control in clear aligner treatment,with the aim to provide a reference for the clinical research and technical development of molar anchorage control in clear aligner treatment.

Pathological dry skin is a disturbing and intractable healthcare burden,characterized by epithelial hy-perplasia and severe itch.Atopic dermatitis(AD)and psoriasis models with complications of dry skin have been studied using single-cell RNA sequencing(scRNA-seq).However,scRNA-seq analysis of the dry skin mouse model(acetone/ether/water(AEW)-treated model)is still lacking.Here,we used scRNA-seq and in situ hybridization to identify a novel proliferative basal cell(PBC)state that exclusively expresses transcription factor CUT-like homeobox 1(Cux1).Further in vitro study demonstrated that Cux1 is vital for keratinocyte proliferation by regulating a series of cyclin-dependent kinases(CDKs)and cyclins.Clinically,Cux1+PBCs were increased in patients with psoriasis,suggesting that Cux1+ PBCs play an important part in epidermal hyperplasia.This study presents a systematic knowledge of the tran-scriptomic changes in a chronic dry skin mouse model,as well as a potential therapeutic target against dry skin-related dermatoses.

Objective:To evaluate the long-term efficacy and safety of a novel self-help inflatable balloon to prevent esophageal stenosis after extensive endoscopic submucosal dissection (ESD).Methods:Patients with early esophageal cancer or precancerous lesions, undergoing ESD in the First Medical Center of Chinese PLA General Hospital from January 2018 to December 2019 were included in the prospective study, who had post-ESD mucosal defect greater than 5/6 of the esophageal circumference and 30-100 mm in length. The self-help inflatable balloon was used to prevent esophageal stenosis after ESD. Mucosal defect of ESD was divided into grade 1 (≥5/6 and less than the whole circumference) and grade 2 (the whole circumference). The incidence of stricture, the time from ESD to the occurrence of stricture, the total number of endoscopic balloon dilations (EBD) or radial incision and cuttings (RIC), and other adverse events were observed.Results:A total of 27 patients met the including criteria with follow-up time of 14-38 months, including 3 patients of grade 1 and 24 of grade 2. The ulcer longitudinal length was 73.7±18.4 mm. The time of wearing balloons was 92.0±20.0 days. The overall frequency of stricture was 18.5% (5/27), and the stricture incidence of patients of grade 2 resection was only 16.7% (4/27). The median time from balloon removal to stricture was 17 days. To treat the stricture, two patients received 3 EBD sessions, and three other patients received 2, 1 and 2 RIC sessions, respectively. No balloon was removed in advance, and none had a perforation or delayed bleeding.Conclusion:The self-help inflatable balloon shows high efficacy and safety in preventing esophageal stenosis in patients with mucosal defect greater than 5/6 of the esophageal circumference and less than 100 mm in length after extensive esophageal ESD.

Objective:To compare the efficacy and safety of endoscopic mucosal resection (EMR), EMR with pre-cutting (EMR-P), endoscopic submucosal dissection (ESD) and ESD with snare (ESD-S) for the treatment of colorectal laterally spreading tumors (LSTs).Methods:Between January 2016 and March 2018, a total of 146 patients with 146 colorectal LSTs undergone endoscopic resection at the first medical center of PLA General Hospital. Data of demographics, treatment information, pathology and follow-up results were retrospectively analyzed.Results:Among the 146 patients, EMR, EMR-P, ESD, and ESD-S were performed in 23, 29, 50 and 44 tumors, respectively. Median tumor diameter was 2.5 cm (ranged 1.2-10.0 cm). The en bloc resection rate of EMR, EMR-P, ESD and ESD-S were 73.9% (17/23), 72.4% (21/29), 96.0% (48/50), and 65.9% (29/44), respectively, with statistical difference ( P<0.001). And the R0 resection rate were 65.2% (15/23), 69.0% (20/29), 94.0% (47/50), and 63.6% (28/44), respectively, with statistical difference ( P=0.002). The en bloc resection rate and R0 resection rate of the ESD group were significantly higher than those of the other three groups (all P<0.05). The difference was not statistically significant in terms of perforation rate [0, 0, 6.0% (3/50), and 9.1% (4/44), respectively, P=0.269] and delayed hemorrhage rate [4.3% (1/23), 0, 2.0% (1/50), and 2.3% (1/44), respectively, P=0.768] among the four groups. Follow-up endoscopy was performed in 117 cases (80.1%) with a median period of 10.0 months (ranged 3.0-26.0 months), and local recurrence was identified in 7 (6.0%) cases. Conclusion:ESD could be the optimal method for the resection of colorectal LSTs, while LSTs smaller than 20 mm can be resected by EMR. EMR-P and ESD-S as modified methods have their respective advantages for the treatment of LSTs.

Objective:To evaluate the clinical value of suspensory incision and suture technique in endoscopic full-thickness resection (EFTR) for muscularis propria tumor of gastric fundus.Methods:A retrospective analysis was performed on the data of 20 patients with muscularis propria tumor in gastric fundus and undergoing EFTR in the First Medical Center of PLA General Hospital from June 2017 to June 2019. Patients were divided into the observation group (9 cases) treated with suspensory incision and suture technique in EFTR and the control group (11 cases) treated with traditional EFTR method. The baseline data and perioperative data of the two groups were analyzed.Results:EFTR was successfully performed on all 20 patients. The tumor size of the observation group and the control group was 10.0 (7.5, 21.0) mm and 14.0 (10.0, 20.0) mm, respectively. The resection time of the two groups was 26.4±6.3 min and 35.5±11.4 min, respectively. The postoperative hospital stay was 6.4±1.0 d and 7.7±1.5 d, respectively. No postoperative delayed bleeding, perforation, or other complications occurred in the two groups.Conclusion:Using suspensory incision and suture technique is safe and effective during EFTR for muscularis propria tumor in gastric fundus, and can reduce operation time. This technique is worth applying in clinic.

Objective To assess the efficacy and safety of autologous skin-grafting surgery ( ASGS) in the prevention of esophageal stenosis after complete circular endoscopic submucosal tunnel dissection ( ESTD) for early esophageal cancer. Methods Between January 2018 and March 2018, five patients with early esophageal cancer underwent complete circular ESTD and ASGS in Chinese PLA General Hospital. The skin-graft survival situation, and occurrence of esophageal stenosis and complications were observed by endoscopy follow-up. Results Complete circular ESTD and ASGS were successfully performed in all 5 patients, and no complications including perforation, bleeding, wound infection or stent migration occurred. The mean skin-graft survival rate was 86. 0％. Four patients did not experience esophageal stenosis over the mean follow-up of 9. 5 months. One patient experienced esophageal stenosis after operation, and underwent endoscopic balloon dilatation. No stenosis occurred in 8 months of follow-up. Conclusion ASGS is a safe and effective method to prevent esophageal stenosis after complete circular ESTD.

Objective: To evaluate optical coherence tomography(OCT)for predicting invasion depth of early esophageal cancer(EEC) and to compare OCT and magnifying endoscopy-narrow band imaging (ME-NBI)in clinical performance. Methods: Twenty-eight patients who were diagnosed with EEC and accepted OCT and ME-NBI before endoscopic submucosal dissection(ESD)were enrolled in this prospective study. On the basis of OCT and ME-NBI images, real-time prediction of EEC invasion depth was conducted. Postoperative pathological results were taken as golden standard to compare the accuracy of OCT and ME-NBI in evaluation of EEC invasion depth. The procedure time and incidence of complications during evaluation process were also analyzed. Results: The overall accuracy of OCT and ME-NBI in predicting invasion depth of 28 EEC patients were 67.9% (19/28) and 75.0% (21/28) respectively, with no significant difference(P>0.05). The accuracy of OCT and ME-NBI in distinguishing lesions located in epithelium/lamina propria mucosa (EP/LPM) lesions were 78.9%(15/19) and 68.4% (13/19), with no significant difference(P>0.05). The procedure time of OCT was significantly shorter than that of ME-NBI (6.0±2.9 min VS 16.3±5.4 min, P<0.001). Conclusion The ability of OCT to predict invasion depth of EEC and distinguish lesions located in the EP/LPM is comparable with that of ME-NBI. Besides, OCT requires shorter procedure time for evaluation.

Objective: To investigate the expression of lncRNA01296 in esophageal cancer (EC) tissues and its effect on the proliferation and migration of EC TE-2 cells. Methods:Atotal of 36 pairs of esophageal cancer tissues and corresponding para-cancerous tissues were collected from EC patients admitted to the Department of Thoracic Surgery,Affiliated Hospital of Chengde Medical College from January 2017 to September 2018. The human normal esophageal epithelial (HEEC) cells and human esophageal cancer cell lines ECA109, TE-1 and TE-2 were cultured. qPCR was used to detect the mRNAexpressions of lincRNA01296, SNRPA(small nuclear ribonucleoproteinA) and NGF (nerve growth factor) in EC tissues and cells. Recombinant lentiviral interference vectoror control vector were used to transfect EC cell lines, as sh-lncRNA01296#1,#2 and Mock groups. WB was used to detect the protein expressions of SNRPAand NGF in transfected cells. MTS assay was used to detect cell proliferation, and Transwell assays were used to detect cell invasion and migration of TE-2 cells after transfection. Results: The mRNAexpressions of lncRNA01296, SNRPAand NGF were significantly increased in esophageal cancer tissues and cell lines (all P<0.01), and these expressions in poorly differentiated TE-2 cells were higher than those in highly differentiated ECA109 and TE-1 cells (all P<0.05). The mRNAexpressions of lncRNA01296 and NGF in sh-lncRNA01296#1 and sh-lncRNA01296#2 groups were significantly lower than those in Mock group (all P<0.01), while the mRNAexpression of SNRPAshowed no statistical difference among three groups (P>0.05). The protein expressions of lncRNA01296 and NGF in sh-lncRNA01296#1 and sh-lncRNA01296#2 groups were significantly lower than those in Mock group (all P<0.01). The relative proliferation ability of cells in sh-lncRNA01296#1 and shlncRNA01296#2 groups was significantly lower than that of Mock group at 48 and 72 h after transfection (P<0.05 or P<0.01). The number of invasive cells was (72.0±6.3), (36.6±4.3) and (33.9±3.7) in Mock, sh-lncRNA01296#1 and sh-lncRNA01296#2 groups, respectively; and the number of migrated cells was (85.2±9.9), (47.5±8.1) and (43.8±6.5), respectively, indicating that the numbers of invasive and migrated cells in sh-lncRNA01296#1 and sh-lncRNA01296#2 groups were significantly less than those in Mock group(all P<0.01). Conclusion: lncRNA01296 can up-regulate SNRPAexpression to promote NGF-mediated proliferation and metastasis of EC cells, which may provide new target for the diagnosis and treatment of esophagealcancer.