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OBJECTIVE@#To investigate the effect of human subcutaneous adipose-derived stem cells (hADSCs) local transplantation on orthodontically induced root resorption (OIRR) and provide theoretical and experimental basis for the clinical application of hADSCs to inhibit OIRR.@*METHODS@#Forty 8-week-old male Sprague Dawley rats were randomly divided into experimental group and control group, with 20 rats in each group, to establish the first molar mesial orthodontic tooth movement (OTM) model of rat right maxillary. The rats in the experimental group were injected with 25 μL of cell suspension containing 2.5×10 5 hADSCs on the 1st, 4th, 8th, and 12th day of modeling, while the rats in the control group were injected with 25 μL of PBS. The rat maxillary models were obtained before and after 7 and 14 days of force application, and 10 rats in each group were killed and sampled after 7 and 14 days of force application. The OTM distance was measured by stereomicroscope, the root morphology of the pressure side was observed by scanning electron microscope and the root resorption area ratio was measured. The root resorption and periodontal tissue remodeling of the pressure side were observed by HE staining and the root resorption index was calculated. The number of cementoclast and osteoclast in the periodontal tissue on the pressure side was counted by tartrate resistant acid phosphatase staining.@*RESULTS@#The TOM distance of both groups increased with the extension of the force application time, and there was no significant difference ( P<0.05). There was no significant difference in OTM distance between the experimental group and the control group after 7 and 14 days of force application ( P>0.05). Scanning electron microscope observation showed that small and shallow scattered resorption lacunae were observed on the root surface of the experimental group and the control group after 7 days of force application, and there was no significant difference in the root resorption area ratio between the two groups ( P>0.05); after 14 days of application, the root resorption lacunae deepened and became larger in both groups, and the root resorption area ratio in the experimental group was significantly lower than that in the control group ( P<0.05). The range and depth of root absorption in the experimental group were smaller and shallower than those in the control group, and the root absorption index in the experimental group was significantly lower than that in the control group after 14 days of force application ( P<0.05). The number of cementoclast in the experimental group was significantly lower than that in the control group after 7 and 14 days of force application ( P<0.05); the number of osteoclasts in the experimental group was significantly lower than that in the control group after 14 days of force application ( P<0.05).@*CONCLUSION@#Local transplantation of hADSCs may reduce the area and depth of root resorption by reducing the number of cementoclasts and osteoclasts during OTM in rats, thereby inhibiting orthodontic-derived root resorption.
Sujet(s)
Rats , Mâle , Humains , Animaux , Rhizalyse/thérapie , Rat Sprague-Dawley , Ostéoclastes , Mouvement dentaire , Cellules souchesRÉSUMÉ
Objective:To evaluate the safety and efficacy of combination of submucosal tunneling endoscopic septum division (STESD) and peroral endoscopic myotomy (POEM) for the treatment of esophageal epiphrenic diverticulum with esophagogastric junction (EGJ) outflow tract disorder.Methods:From October 2017 to August 2021, 6 patients with esophageal epiphrenic diverticulum complicated with EGJ outflow tract disorder receiving combination of STESD and POEM in the Endoscopic Center of the First Affiliated Hospital of Nanjing Medical University were enrolled. The clinical characteristics of the patients were retrospectively analyzed, which included the ratio of male to female, age, course of disease, length of diverticula, location, whether with multiple diverticula, type of EGJ outflow tract disorders, whether the endoscopic technique was successful (the completion of all steps under endoscopy, including tunnel establishment and diverticulum cristae, lower esophageal sphincter and cardia muscularis propria incision); operation time, changes in the severity of clinical symptoms before and after operation (including weight loss, dysphagia, retrosternal pain, and reflux assessed using the Eckardt score), intraoperative and postoperative complications, and follow-up, including whether achieved clinical success (complete or nearly complete improvement of dysphagia, vomiting after eating, retrosternal pain, regurgitation, weight loss, no need for repeat endoscopic intervention during follow-up) and adverse events. Descriptive methods were used for statistical analysis.Results:The male to female ratio of the 6 patients was 3 to 3, the median age was 69.2 years old, and the median disease course was 92.3 months. The median length of the diverticula was 47.5 mm. Diverticulum was located in the right wall of esophagus in 4 cases and in the left wall of esophagus in 2 cases, of which 1 patient was multiple diverticulum of esophagus. Achalasia was found in 5 cases, and EGJ outflow obstruction was found in 1 case. All the 6 patients successfully completed combination of STESD and POEM, and all achieved successful edoscopic technique. The operation time (range) was 55 min (40 to 70 min). Clinical symptom Eckardt score (range) before and after operation was 9.0 (7.0 to 11.0) and 1.3 (1.0 to 2.0), respectively. After operation the clinical symptom improved compared with that before operation. There were no delayed bleeding, perforation, infection, subcutaneous emphysema and other complications and severe adverse events. Six patients were all cured and discharged. Follow-up period was 1 to 50 months. The symptoms of dysphagia, vomiting after eating, retrosternal pain, regurgitation and weight loss were all significantly improved compared with those before operation. There were no severe adverse events and all achieved clinical success.Conclusions:Combination of STESD and POEM is safe and effective in the treatment of esophageal epiphrenic diverticulum with EGJ outflow tract disorder, and has good short-term and long-term effects.
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It is a thorny problem of modern medicine that the in-stent restenosis (ISR) after percutaneous coronary intervention (PCI). Combining with the etiology and pathogenesis of TCM, Professor LIU Zhong-yong believes that the root cause of ISR after PCI is the deficiency syndrome: menstruation gradually dying up, the heart yang qi deficiency; and the direct cause is excess syndrome: endogenous turbidity syndrome, heart vessel blockage. The cause for the formation of turbidity syndrome is cold, phlegm, blood stasis, poison, and dampness. Depending on the clinical manifestations, five kinds of syndromes were divided: cold turbidity stagnation, phlegm turbidity resistance, blood stasis blockage veins, poison turbidity, and dampness turbidity resistance. Professor LIU Zhong-yong also proposed relevant treatment for both symptoms and root causes, which provided new ideas and experience in the integrated TCM and Western medicine for ISR after PCI.
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Objective:To study the chemical constituents in Blumea aromatica ( Wall. ) DC. Methods:The compounds were iso-lated by silica gel, Sephadex LH-20 column chromatography and TLC. The structures were elucidated by physicochemical properties and spectral analysis. Results:Ten compounds were obtained and elucidated as oleic acid (1), linoleic acid (2), quercetin (3), luteolin (4), naringenin (5), hesperetin (6), gallic acid (7), β-sitosterol (8), daucosterol (9) and hesperidin (10). Conclu-sion:Compounds 1, 2, 5, 6, 7 and 10 are isolated from the plant for the first time.
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Objective:To investigate the volatile constituents in Folium isatidis. Methods:The volatile constituents from Folium isatidis were analyzed by head-space solid micro-extraction coupled with GC-MS for the first time. Results: Thirty-five compounds (89. 95%) were identified from the volatile constituents in Folium isatidis. The main volatile constituents of Folium isatidis were 6, 10, 14-trimethyl-2-pentadecanone (6. 32%), nonanal (5. 99%), phenethyl isothiocyanate (5. 79%) and palmitic acid (5. 62%). Conclusion:Palmitic acid and benzyl alcohol may be the main effective constituents in Folium isatidis.
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Objective To investigate the effects of hypoxia-inducible factor (HIF)-1α on glycolysis of human esophageal squamous carcinoma cells and the possible mechanism.Methods TE13 and Eca109 cells were cultured under normal oxygen (20％O2) and hypoxia (1％O2) conditions.The hypoxia was duration 6 hours,12 hours,24 hours and 48 hours.HIF-1α gene was stable silented by RNA interference method and TE13/small interfering HIF cells and Eca109/siHIF cells were obtained.The cell culture condition and time was same as TE13 and Eca109 cells.The changes of HIF-1α expression were detected by Western-blot.The changes of lactic acid concentration in cell culture supernatant were determined by Spectrophotometry.The changes of glucose transporter-1 (GLUT-1) and lactic dehydrogenase A (LDHA) expression at mRNA level were examined by realtime polymerase chain reaction.The changes of GLUT-1 and LDHA expression at protein level were tested by Western blot.Using t or t' test to analyze the effects of hypoxia duration on HIF-1αexpression at protein level.One-way ANOVA was applied for the difference analysis between the groups.Results In TE13 and Eca109 cells,the HIF-1α expression significantly increased under hypoxia condition and reached the peak at 12 hour (t=6.11,8.31; both P＜0.05).The lactic acid secretion of TE13/siHIF cells and Eca109/siHIF cells was (1.24±0.33) and (1.28±0.37) mmol/L,which significantly decreased when compared with TE13 and Eca109 cells [(3.25±1.34) and (4.91±1.69) mmol/L,t=2.53,3.59,both P＜0.05].The lactic acid secretion of TE13 and Eca109 cells significantly increased after hypoxia [(6.48±1.73) and (8.02± 1.95) mmol/L,t=2.715,2.050,both P＜0.05].There was no significant lactic acid secretion in TE13/siHIF cells and Eca109/siHIF cells after hypoxia (P ＞ 0.05).The expressions of GLUT-1 and LDHA at mRNA level were significantly suppressed in TE13/siHIF cells and Eca109/siHIF cells (normal oxygen:t=6.98,3.92,7.25,3.67,all P＜0.05).The expression of GLUT-1 at protein level remarkably weaked (normal oxygen:t=4.57、16.56,hypoxia:t=6.19、6.09,all P＜0.05),while the expression of LDHA at protein level slightly decreased (P＞0.05).Conclusions The level of glycolysis can be lowered by suppression HIF-1α expression in human esophageal squamous carcinoma cells.The pathway may be involved in the suppression of GLUT-1 and LDHA expression.Except for HIF-1α,there may be other regulating factors in LDHA protein expression at same time.
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Objective To investigate the changes of hypoxia-inducible factor(HIF)-1α and hexokinase-Ⅱ(HK-Ⅱ)expression in human esophageal squamous cell carcinoma and its effect in glycolysis.Methods TE13 cells and Eca109 cells were cultured under hypoxic condition(1 ％O2)for different hypoxic time(6,12,24 and 48 hours).Cells cultured under normal oxygen condition(20％O2)were set as control.The changes of HIF-1α and HK-Ⅱ expressions at protein level were detected by Western blot.HIF-1α genes were specifically silenced with RNA interference technology(RNAi),and then the changes of HIF-1α and HK-Ⅱ expression were determined by realtime PCR and Western blot.Under normal oxygen and hypoxic condition,the changes of lactic acid concentration in cell culture medium were detected by spectrophotometric method.Results Under hypoxic condition,the expression of HIF-1α and HK-Ⅱ gradually increased as hypoxic time extended(P＜0.05),reached a peak at 12h and then gradually decreased as time extended.Compared with that under normal oxygen condition,the expression of HK-Ⅱ in TE13 cells and Eca109 cells significantly increased under hypoxic condition(P＜0.05),which was more significant after 12 hours hypoxia.The result of realtime PCR indicated that under normal oxygen condition the expression of HIF-1α at RNA level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference(P＜0.05).The expression of HK-Ⅱ at RNA level was consistent with the result of HIF-1α.Under normal and hypoxia condition,the expression of HK-Ⅱ at protein level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).The lactic acid secretion of TE13 cells and Eca109 cells under hypoxia condition(14.707 ± 3.594 and 15.062 ±3.901)was higher than that under normal oxygen condition(6.070±1.839 and 6.891±1.592,P＜0.05).The lactic acid secretion of TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).Conclusion The expressions of HIF-1α and HK-Ⅱ in human esophageal squamous cell carcinoma significantly increased under hypoxia conditions.The expression of HK-Ⅱ is closely correlated with lactic acid concentration and HIF-1α expression.HIF-1α may affect cell glycolysis through HK-Ⅱ.
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Objective To investigate the inhibitory effect of blocking PI3K/AKT pathway by wortmannin on hypoxia-inducible factor 1α (HIF-1α) and the effect on the expression of glycolysis associated genes in human esophageal carcinoma cell lines TE1 and TE13,and to analyze the relation between PI3K/AKT-HIF-1α pathway and glycolysis in esophageal carcinoma cells. Methods Esophageal carcinoma cell lines TE1 and TE13 pretreated with wortmannin (2 μmol/L) were incubated under normoxic and hypoxic conditions.And each cell line was divided into four groups.The expression of HIF-1α and glycolysis associated genes GLUT-1,LDHA and HK-Ⅱ at protein level were measured by.Western blot.The expression of HIF-1α,GLUT-1,LDHA and HK-Ⅱ at mRNA level was determined by real-time PCR. The activities of LDH and HK-Ⅱ and lactic acid (LA)concentration in the culture supernatant were tested with spectrophotometer method.Results Under normoxic condition,HIF-1α was expressed in TE1 cells and the expression of HIF-1α was inhibited by wortmannin (2 μmol/L),the most significant inhibitory effect was at 12 hours,therefore 12 hours was selected for the subsequent hypoxia experiment.Compared with untreated cells,the expression of HIF-1α、HK-Ⅱ 、GLUT-1、LDH-A at protein level significantly decreased in TE1 and TE13 cells after pretreated with wortmannin (P ＜ 0.05),and the expression of HIF-1α、HK-Ⅱ at mRNA level significantly decreased (P＜ 0.05).Under normoxic and hypoxic conditions,the HK-Ⅱ and LDH activities in TE1 and Te13 esophageal carcinoma cells significantly decreased after treated with wortmannin compared with untreated cells (P＜0.05).Under hypoxia condition,the enzyme activity increased in untreated cells (P＜ 0.05). Under normoxic and hyp0xic conditions,the lactic acid concentration in the culture supernatant obviously decreased in cells treated with wortmannin compared with untreated cells (P＜ 0.05). Under hypoxia condition,lactic acid concentration increased in wortmannin treated cells (P ＜ 0.05). Conclusions Under normoxic and hypoxic conditions,wortmannin decrease lactic acid concentration through inhibiting the expression of HIF-1α and glycolysis associated genes, which indicate PI3K/AKT-HIF-1α pathway was closely related to glycolysis in esophageal carcinoma cells.
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Objective To evaluate the efficacy and safety of covered metallic stents in the management of proximal esophageal strictures.Methods Retrievable covered self-expanding metallic stents were inserted individually in 43 patients with malignant strictures of the proximal esophagus.Main outcome measures included improvement of dysphagia,sealing of fistula and complications were evaluated.Results A total of 45 stents were placed in 43 patients,with a success rate of 95.6％ (43/45) at the first stent placement attempt.The dysphagia score improved significantly from 3.3 ± 0.6 to 1.8 ± 0.9 (P ＜ 0.001),with 18 having an excellent response and 25 having a good response.Improvement rate of dysphagia was 95.6％ (43/45).Fistula sealing was achieved in 7 of 9 patients (77.8％).Early complications included severe pain (n =5),mediastinal emphysema (n =1),moderate fever (n =9) and collapse of the proximal end of the stent (n =1),while late complications included migration (n =3) and tissue overgrowth (n =6).Conclusion Retrievable covered metallic stent placement is effective and safe for the palliation of obstructive dysphagia and sealing of fistulas in malignant proximal esophageal strictures.
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Objective:To study the effect of GABA and A receptor agonist THIP on proliferation and apoptosis of cytokine-induced killer (CIK) cells.Methods:CIK cells were cultured by routine method,and then treated with different concentrations of GABA and THIP.The proliferation of CIK cells were investigated by MTT assay.The apoptosis,cell cycle and immunophenotype were investigated by flow cytometry.Results:GABA could inhibit the proliferation of CIK cells in a dose-dependent manner and affect the distribution of cell cycle of CIK cells(P