BACKGROUND:Adipose tissue-derived mesenchymal stem cells release a large amount of exosomes to participate in various pathophysiological processes,but the impact and precise mechanism of exosomes derived from adipose tissue-derived mesenchymal stem cells on autophagy of hepatic stellate cells have not been fully elucidated.OBJECTIVE:To explore the targeted regulatory effect and molecular mechanism of adipose tissue-derived mesenchymal stem cell-derived exosomes on autophagy of hepatic stellate cells through miR-15a-5p.METHODS:Adipose tissue was collected from inguinal region of 8-week male C57BL/6 mice.Adipose tissue-derived mesenchymal stem cells were extracted by collagenase digestion.Adipose tissue-derived mesenchymal stem cell-derived exosomes were extracted by ultracentrifugation.Mouse liver tissue was obtained,and hepatic stellate cells were isolated and extracted using collagenase perfusion digestion and density gradient centrifugation.The experiment was divided into two groups.In control group,hepatic stellate cells were cultured alone for 48 hours.In the exosome group,hepatic stellate cells were co-cultured with adipose tissue-derived mesenchymal stem cell-derived exosomes for 48 hours.The effects of exosomes on hepatic stellate cell proliferation,activation,autophagy,and expression of fibrosis markers were detected by western blot assay,RT-qPCR,and immunofluorescence staining.RT-qPCR and western blot assay were used to detect the effect of exosomes on the mRNA and protein expression of miR-15a-5p and the downstream signaling pathway Bcl-2,Beclin-1,and Rubicon in hepatic stellate cells.RESULTS AND CONCLUSION:(1)Compared with the control group,the ratio of autophagy markers LC3-Ⅱ expression decreased,the number of autophagosome was also significantly decreased,and the intracellular lipid droplets were regenerated,simultaneously,cell volume diminished with the weakening of proliferation in hepatic stellate cells of the exosome group,indicated that the hepatic stellate cell activation was significantly inhibited.(2)Compared with the control group,the expressions of α-smooth actin and type Ⅰ collagen were significantly decreased(P＜0.01),and the expression of miR-15a-5p was significantly increased in hepatic stellate cells of the exosome group(P＜0.01).At the same time,the expression of its downstream target gene Bcl-2 was significantly decreased(P＜0.01),while the expressions of autophagy genes Beclin-1 and Rubicon were significantly increased in hepatic stellate cells of the exosome group(P＜0.01).The results indicate that adipose tissue-derived mesenchymal stem cell-derived exosomes inhibits the expression of Bcl-2 in hepatic stellate cells by targeting miR-15a-5p and increases the expression of downstream autophagy genes Beclin-1 and Rubicon,thereby inhibiting the autophagy of hepatic stellate cells.

BACKGROUND:Adipose tissue-derived mesenchymal stem cells release a large amount of exosomes to participate in various pathophysiological processes,but the impact and precise mechanism of exosomes derived from adipose tissue-derived mesenchymal stem cells on autophagy of hepatic stellate cells have not been fully elucidated.OBJECTIVE:To explore the targeted regulatory effect and molecular mechanism of adipose tissue-derived mesenchymal stem cell-derived exosomes on autophagy of hepatic stellate cells through miR-15a-5p.METHODS:Adipose tissue was collected from inguinal region of 8-week male C57BL/6 mice.Adipose tissue-derived mesenchymal stem cells were extracted by collagenase digestion.Adipose tissue-derived mesenchymal stem cell-derived exosomes were extracted by ultracentrifugation.Mouse liver tissue was obtained,and hepatic stellate cells were isolated and extracted using collagenase perfusion digestion and density gradient centrifugation.The experiment was divided into two groups.In control group,hepatic stellate cells were cultured alone for 48 hours.In the exosome group,hepatic stellate cells were co-cultured with adipose tissue-derived mesenchymal stem cell-derived exosomes for 48 hours.The effects of exosomes on hepatic stellate cell proliferation,activation,autophagy,and expression of fibrosis markers were detected by western blot assay,RT-qPCR,and immunofluorescence staining.RT-qPCR and western blot assay were used to detect the effect of exosomes on the mRNA and protein expression of miR-15a-5p and the downstream signaling pathway Bcl-2,Beclin-1,and Rubicon in hepatic stellate cells.RESULTS AND CONCLUSION:(1)Compared with the control group,the ratio of autophagy markers LC3-Ⅱ expression decreased,the number of autophagosome was also significantly decreased,and the intracellular lipid droplets were regenerated,simultaneously,cell volume diminished with the weakening of proliferation in hepatic stellate cells of the exosome group,indicated that the hepatic stellate cell activation was significantly inhibited.(2)Compared with the control group,the expressions of α-smooth actin and type Ⅰ collagen were significantly decreased(P＜0.01),and the expression of miR-15a-5p was significantly increased in hepatic stellate cells of the exosome group(P＜0.01).At the same time,the expression of its downstream target gene Bcl-2 was significantly decreased(P＜0.01),while the expressions of autophagy genes Beclin-1 and Rubicon were significantly increased in hepatic stellate cells of the exosome group(P＜0.01).The results indicate that adipose tissue-derived mesenchymal stem cell-derived exosomes inhibits the expression of Bcl-2 in hepatic stellate cells by targeting miR-15a-5p and increases the expression of downstream autophagy genes Beclin-1 and Rubicon,thereby inhibiting the autophagy of hepatic stellate cells.

OBJECTIVEThis research aimed to observe the expression of heat shock protein 27 (HSP27) in human gingival fibroblasts (HGFs) cell injury induced by different concentrations of cigarette smoke extract (CSE).

METHODSThe third to eighth generations of cultured HGFs were treated with serially diluted CSE of different concentrations (0, 2.5%, 5.0%, 12.5%, 25.0%, 50.0%). Wound-healing assay was performed to determine the migration of HGFs, and Western blot was used to determine the expression of HSP27.

RESULTSThe migration capability of HGFs weakened with the increase of CSE concentration. HSP27 expression was negative in normal HGFs but positive in CSE-intervened HGFs in a concentration-dependent manner.

CONCLUSIONHSP27 concentration increased in the CSE-induced injury of HGFs. This finding suggests that HSP27 plays an important role in CSE-induced epithelial injury.

Cells, Cultured ; Fibroblasts ; Gingiva ; HSP27 Heat-Shock Proteins ; Humans ; Smoke ; Tobacco ; Wound Healing

Objective This research aimed to observe the expression of heat shock protein 27 (HSP27) in human gingival fibroblasts (HGFs) cell injury induced by different concentrations of cigarette smoke extract (CSE). Methods The third to eighth generations of cultured HGFs were treated with serially diluted CSE of different concentrations (0, 2.5%, 5.0%, 12.5%, 25.0%, 50.0%). Wound-healing assay was performed to determine the migration of HGFs, and Western blot was used to determine the expression of HSP27. Results The migration capability of HGFs weakened with the increase of CSE concentration. HSP27 expression was negative in normal HGFs but positive in CSE-intervened HGFs in a concentration-dependent manner. Conclusion HSP27 concentration increased in the CSE-induced injury of HGFs. This finding suggests that HSP27 plays an important role in CSE-induced epithelial injury.

Objective To investigate the influence of diabetes mellitus (DM) on left ventricular(LV) remodeling in patients with acute ST-segment elevation myocardial infarction (STEMI) treated by primary percutaneous coronary intervention (PCI) within 12 hours of symptom onset. Methods Four hundred and fifty-one consecutive patients with acute STEMI treated by primary PCI were prospectively enrolled in the current study. Baseline, angiographic and PCI features and prevalence of LV remodeling at one-week during hospitalization and 6-month clinical follow-up by two-dimensional echocardiography were compared between 93 diabetic and 358 non-diabetic patients. Results Despite similar baseline clinical and angiographic characteristics, symptom-to-door time was longer (399±106 min vs. 321±116 min, P=0.006) and prevalence of multivessel disease was higher (65.6%vs. 51.7%, P=0.02) in diabetic patients. More patients in diabetic group had LV remodeling at 6-month clinical follow-up (29.0%vs. 17.3%, P=0.01), and DM was an independent predictor of LV remodeling (RR 2.1, 95%CI 1.31-4.79, P=0.02). The rate of rehospitalization due to heart failure did not differ between diabetic and non-diabetic patients (12.9%vs. 8.1%, P=0.15), however, more adverse events occurred in patients with LV remodeling comparing to those without LV remodeling (25.8% vs. 6.6%, P < 0.001). Conclusions Diabetic patients with STEMI often have an increased risk of LV remodeling after treated by primary PCI. Thus, comprehensive therapeutic strategy for diabetic patients presented with STEMI is required considering the poor prognosis of these patients with LV remodeling.

Objective To study the effect of computerized pressure hemostat on lower limb deep vein thrombosis in high-risk elderly patients.Methods From September to December 2013,36 orthopedic hospitalized patients who received operations were divided into the hemostat group and the non-hemostat group with 18 patients in each group.D-dimer changes and lower limb deep vein thrombosis were observed in both groups.Results D-dimer was significantly increased in both groups compared with that before operation,but the extent of elevation in the non-hemostat group was lower than that in the hemostat group.D-dimer and cases of lower limb deep vein thrombosis in the hemostat group was higher than those in the non-hemostat group after operation,which showed significant difference.Conclusions The use of computerized pressure hemostat will increase the risk of lower limb deep vein thrombosis in elderly patients,so the technical operation procedures should be strictly enforced accompany with safely use of computerized pressure hemostat.

Objective To analyse and compare the effects and safety of early use (in emergency room, intravenous loading followed by infusion) with bolus injection during primary PCI of tirofiban, on post-procedural TIMI flow and 30d clinical outcomes. Methods Seven hundred and seven patients with acute STEMI treated by primary PCI in Ruijin hospital were retrospectively and enrolled screened. Among them, 86 patients with single bolus intra-coronary injection of tirofiban (25 μg/kg) during the procedure were served as observation group. Baseline, angiographic, PCI features and rate of major adverse cardiac events (MACE) at 30 d follow-up were compared with those received early intravenous infusion of tirofiban (10ug/kg bolus followed by 0.15μg/(kg·min) intravenous infusion)(control group, n=239). Results Compared with control group, patients in observation group were older[(63.8±11.4) vs. (57.9±8.8), P=0.01], had higher prevalence of hypertension (58.6%vs. 51.0%, P=0.005), multivessel disease (57.0%vs. 34.3%, P<0.001), and female in gender (40.7%vs. 25.1%, P=0.006). Post-procedural TIMI flow in culprit vessel and TMP grade were comparable between the two groups (P=0.66 and P=0.48, respectively). Reduction in TIMI minimal bleeding events were found in the observation group (2.3%vs. 9.6%, P=0.03). MACE free survival rate at 30d clinical follow-up was similar between the two groups (P=0.48). Conclusions Single bolus intra-coronary injection of tirofiban exerts similar effects in post-procedural TIMI flow, TMP grade in culprit vessel and 30d clinical outcomes compared with early use in emergency room with intra-venous loading and infusion, nevertheless, intra-coronary injection resulted in significantly reduced TIMI minimal bleeding events. Prospective, randomized clinical study is mandatory to prove our current results.

OBJECTIVETo observe the short-term clinical effectiveness of bone ring graft technique and to summarize the key points of related surgical operation to provide comprehensive clinical guidelines.

METHODSFifteen patients with severe alveolar bone absorption were selected to receive bone ring grafting and immediate dental implant. Final fixed prostheses were cemented five months after initial implantation. Cone beam CT scans were conducted on all subjects before the procedure, as well as four months post-operation to evaluate alveolar bone height and level of bone height and absorption around the implants. Four to six months after prosthesis installation, each implant's Jemt classification, gingiva attachment, and probing depth (PD) were analyzed. The difference of PD between implants and adjacent teeth, as well as the difference of the bone absorption between labial and lingual sides, was compared. The survival rate of the bone ring and the retention rate of implants were calculated. Complications and patient satisfaction were also investigated.

RESULTSBone graft survival rate was 94.4% and dental implantation retention rate was 100% four months post-operation. Average bone level increase was (6.06 +/- 1.06) mm, average bone absorption was (1.33 +/- 0.84) mm, and average bone thickness at the neck of the dental implant body was (6.94 +/- 0.73) mm. Approximately 4 to 6 months after crown restoration, average bone level increase was (5.62 +/- 1.03) mm, average bone absorption was (1.51 +/- 1.02) mm, and average bone thickness at the neck of the dental implant body was (6.77 +/- 0.72) mm. The PD around the implant body and the adjacent teeth was statistically insignificant. No major post-operative complication was observed, restorations were successful, and patient satisfaction level was high.

CONCLUSIONBone ring graft technique and immediate dental implantation are relatively simple to perform, and these techniques facilitate reduction in required treatment time. Short-term effect is reliable and satisfactory, whereas long-term outcomes require further follow up and study.

Alveolar Bone Loss ; Bone Transplantation ; Crowns ; Dental Implantation, Endosseous ; Dental Implants ; Dental Prosthesis Design ; Dental Prosthesis, Implant-Supported ; Dental Restoration Failure ; Humans

BACKGROUNDCollaterals to occluded infarct-related coronary arteries (IRA) have been observed after the onset of acute ST-elevation myocardial infarction (STEMI). We sought to investigate the impact of early coronary collateralization, as evidenced by angiography, on myocardial reperfusion and outcomes after primary percutaneous coronary intervention (PCI).

METHODSAcute procedural results, ST-segment resolution (STR), enzymatic infarct size, echocardiographic left ventricular function, and major adverse cardiac events (MACE) at 6-month follow-up were assessed in 389 patients with STEMI undergoing primary PCI for occluded IRA (TIMI flow grade 0 or 1) within 12 hours of symptom-onset. Angiographic coronary collateralization to the occluded IRA at first contrast injection was graded according to the Rentrop scoring system.

RESULTSLow (Rentrop score of 0 or 1) and high (Rentrop score of 2 or 3) coronary collateralization was detected in 329 and 60 patients, respectively. Patients with high collateralization more commonly had prior stable angina and right coronary artery occlusion, but less often had left anterior descending artery occlusion. At baseline, these patients presented with less extent of ST-segment elevation and lower serum levels of creatine kinase myocardial band (CK-MB) and cardiac troponin I (cTnI). Procedural success rate, STR, corrected TIMI flame count, and area under the curve of CK-MB and cTnI measurements after the procedure were similar between patients with high collateralization and those with low collateralization (for all comparisons P > 0.05). There were no differences in left ventricular ejection fraction and rates of MACE at 6 months according to baseline angiographic collaterals to occluded IRA.

CONCLUSIONSIn patients with acute STEMI undergoing primary PCI within 12 hours of symptom-onset, coronary collateralization to the occluded IRA was influenced by clinical and angiographic features. Early recruitment of collaterals limits infarct size at baseline, but has no significant impact on myocardial reperfusion after the procedure and subsequent left ventricular function and clinical outcomes.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Angioplasty, Balloon, Coronary ; Coronary Angiography ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnostic imaging ; therapy ; Treatment Outcome ; Young Adult

Objective To observe the short-term clinical effectiveness of bone ring graft technique and to summarize the key points of related surgical operation to provide comprehensive clinical guidelines. Methods Fifteen patients with severe alveolar bone absorption were selected to receive bone ring grafting and immediate dental implant. Final fixed prostheses were cemented five months after initial implantation. Cone beam CT scans were conducted on all subjects before the procedure, as well as four months post-operation to evaluate alveolar bone height and level of bone height and absorption around the implants. Four to six months after prosthesis installation, each implant’s Jemt classification, gingiva attachment, and probing depth (PD) were analyzed. The difference of PD between implants and adjacent teeth, as well as the difference of the bone absorption between labial and lingual sides, was compared. The survival rate of the bone ring and the retention rate of implants were calculated. Complications and patient satisfaction were also investigated. Results Bone graft survival rate was 94.4% and dental implantation retention rate was 100% four months post-operation. Average bone level increase was (6.06±1.06) mm, average bone absorption was (1.33±0.84) mm, and average bone thickness at the neck of the dental implant body was (6.94± 0.73) mm. Approximately 4 to 6 months after crown restoration, average bone level increase was (5.62±1.03) mm, average bone absorption was (1.51±1.02) mm, and average bone thickness at the neck of the dental implant body was (6.77±0.72) mm. The PD around the implant body and the adjacent teeth was statistically insignificant. No major post-operative complication was observed, restorations were successful, and patient satisfaction level was high. Conclusion Bone ring graft technique and immediate dental implantation are relatively simple to perform, and these techniques facilitate reduction in required treatment time. Short-term effect is reliable and satisfactory, whereas long-term outcomes require further follow up and study.