Periodontitis is a common oral disease caused by bacteria coupled with an excessive host immune response. Stem cell therapy can be a promising treatment strategy for periodontitis, but the relevant mechanism is complicated. This study aimed to explore the therapeutic potential of mitochondria from human embryonic stem cell-derived mesenchymal stem cells (hESC-MSCs) for the treatment of periodontitis. The gingival tissues of periodontitis patients are characterized by abnormal mitochondrial structure. Human gingival fibroblasts (HGFs) were exposed to 5 μg/mL lipopolysaccharide (LPS) for 24 h to establish a cell injury model. When treated with hESC-MSCs or mitochondria derived from hESC-MSCs, HGFs showed reduced expression of inflammatory genes, increased adenosine triphosphate (ATP) level, decreased reactive oxygen species (ROS) production, and enhanced mitochondrial function compared to the control. The average efficiency of isolated mitochondrial transfer by hESC-MSCs was determined to be 8.93%. Besides, a therapy of local mitochondrial injection in mice with LPS-induced periodontitis showed a reduction in inflammatory gene expression, as well as an increase in both the mitochondrial number and the aspect ratio in gingival tissues. In conclusion, our results indicate that mitochondria derived from hESC-MSCs can reduce the inflammatory response and improve mitochondrial function in HGFs, suggesting that the transfer of mitochondria between hESC-MSCs and HGFs serves as a potential mechanism underlying the therapeutic effect of stem cells.
Humans ; Gingiva/cytology* ; Fibroblasts/metabolism* ; Mitochondria/physiology* ; Mesenchymal Stem Cells/cytology* ; Animals ; Periodontitis/therapy* ; Mice ; Reactive Oxygen Species/metabolism* ; Inflammation ; Lipopolysaccharides ; Human Embryonic Stem Cells/cytology* ; Cells, Cultured ; Adenosine Triphosphate/metabolism* ; Male

Objective:To explore the mechanism of a novel BMX splicing variant induced epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) gefitinib resistance in lung cancer.Methods:Stable transgenic cell line PC9-BMXΔN and HCC827-BMXΔN were constructed by lentivirus infection of PC9 and HCC827 cells carrying EGFR mutation. The cells were divided into PC9-Vec group (PC9 cells transfected with empty vector), PC9-BMX group (PC9 cells stably expressing BMX), PC9-BMXΔN group (PC9 cells stably expressing BMXΔN) and HCC827-Vec group (HCC827 cells transfected with empty vector), HCC827-BMXΔN group (HCC827 cells stably expressing BMXΔN). Quantitative real-time PCR was used to detect the expression levels of mRNA. The protein expression levels in each group were detected by Western blotting. The cells in the PC9-Vec group and PC9-BMXΔN group were treated with 0, 0.01, 2.00, 50.00, 100.00, 200.00 nmol/L and 2.00, 4.00 μmol/L gefitinib. The cells in the HCC827-Vec group and HCC827-BMXΔN group were treated with 0, 0.01, 1.00, 10.00, 100.00 nmol/L and 1.00 μmol/L gefitinib. MTT method was used to detect cell viabilities.Results:The PC9-BMXΔN cells were scattered and showed a fibroblast-like morphology. Compared with the PC9-Vec cells, the relative expression levels of fibronectin, N-cadherin, vimentin, Snail, Slug and TWIST 2 mRNA in PC9-BMXΔN cells were up-regulated. Compared with the PC9-Vec cells and PC9-BMX cells, the expression levels of fibronectin and vimentin protein in PC9-BMXΔN cells were up-regulated; while the expression level of E-cadherin protein in PC9-BMXΔN cells was significantly down-regulated. Compared with the PC9-Vec cells, the cell viabilities of PC9-BMXΔN cells treated with 0.01 nmol/L [(99.11±2.16)% vs. (91.29±1.91)%, t=-4.701, P=0.011], 2.00 nmol/L [(80.41±1.48)% vs. (63.36±2.14)%, t=-11.324, P<0.001], 50.00 nmol/L [(80.83±5.38)% vs. (60.22±3.61)%, t=-5.507, P=0.005], 100.00 nmol/L [(75.54±3.46)% vs. (59.93±1.91)%, t=-6.836, P=0.002], 200.00 nmol/L [(77.57±6.53)% vs. (56.70±2.88)%, t=-5.064, P=0.007], 2.00 μmol/L [(70.22±3.45)% vs. (53.14±0.89)%, t=-8.309, P=0.001], 4.00 μmol/L [(68.66±4.67)% vs. (52.30±2.59)%, t=-4.882, P=0.008] gefitinib were significantly increased, with statistically significant differences. Similarly, compared with the HCC827-Vec cells, the cell viabilities of HCC827-BMXΔN cells treated with 1.00 nmol/L [(64.36±2.49 )% vs. (47.13±4.21)%, t=-7.067, P=0.019], 10.00 nmol/L [(63.25±5.87)% vs. (43.28±2.95)%, t=-5.267, P=0.006], 100.00 nmol/L [(49.47±5.74)% vs. (37.12±4.92)%, t=-2.830, P=0.047], 1.00 μmol/L [(49.05±3.34)% vs. (32.06±4.73)%, t=-5.073, P=0.007] gefitinib were significantly increased, with statistically significant differences. Gefitinib treatment could significantly inhibit the expression levels of p-EGFR and p-ERK1/2 both in PC9-Vec cells, PC9-BMX cells and PC9-BMXΔN cells. Compared with the PC9-Vec cells and PC9-BMX cells, the expression level of p-EGFR in PC9-BMXΔN cells was significantly increased after gefitinib treatment for 8 h (0.91±0.04 vs. 0.81±0.04 vs. 0.80±0.05, all P<0.05); the expression levels of p-ERK1/2 in PC9-BMXΔN cells were significantly increased after gefitinib treatment for 2 h (0.64±0.06 vs. 0.38±0.12 vs. 0.37±0.14), 4 h (1.28±0.06 vs. 1.08±0.06 vs. 1.11±0.07), and 8 h (0.75±0.04 vs. 0.55±0.05 vs. 0.60±0.07), with statistically significant differences (all P<0.05). Conclusion:BMXΔN is involved in EGFR-TKI gefitinib resistance in lung cancer, which may be achieved by inducing cells to undergo epithelial-mesenchymal transition and activating the ERK/MAPK signaling pathway.

Objective To investigate the effects of the angle of external jugular vein on the incidence of PICC complications.Methods A total of 212 patients with external jugular vein tube from August 2013 to November 2015 were divided into acute angle type (100 cases) and obtuse angle type (112 cases) according to the angle of external jugular vein by the X-ray. PICC complications of two groups were evaluated.Results There were 4 cases of mechanical phlebitis, 2 cases of thrombosis and 0 case of infection in the obtuse angle type group, while in the acute angle type group there were 19 cases of mechanical phlebitis, 9 cases of thrombosis and 4 cases of infection. Statistically significant differences were observed between two groups (χ2=13.01, 5.58, 4.57;P<0.05).Conclusions External jugular vein with obtuse angle type has lower incidence of PICC complications.

ObjectiveTo analyze the expression of osteopontin (OPN) and interleukin-17 (IL-17)and study the roles in the pathogenesis of chronic severe hepatitis B.MethodsTwenty patients of acute on chronic liver failure were collected from the infection disease in-patients department of the First Affiliated Hospital of Medical College,Zhejiang University from 2009 to 2010,and 40 health controls were from medical examination center during the same period.Meanwhile,Balb/C mice were used for ConA injection to induce fulminant hepatitis and the plasma,serum and liver tissue of mice were collected.OPN and IL-17concentrations were measured using ELISA kits.PBMC were separated and cultured with anti-TNF-α or TNF-α.Supernatants were collected for analysis of OPN and IL-17.Differences between groups were evaluated by using a Student's t test and the relationship between IL-17 and OPN were evaluated by Pearson correlation.ResultsIn chronic severe hepatitis B group,levels of OPN and IL-17 were markedly higher than those of healthy control,respectively.(42.4 ± 12.9 vs 10.6 ±4.8 pg/ml; 1460.1 ±523.1 vs 472.8 ±67.5 ng/ml) ( t =2.387,3.570,P ＜ 0.05).The level of OPN in blood and liver reached peak at 6 hours at 12 hours after ConA injection,respectively.The level of IL-17 in blood and liver was significantly increased after ConA injection.IL-17 were positive correlated with OPN levels (R2 =0.582,P =0.026).TNF-α can increase the level of OPN secreted by lymphocytes.ConclusionsOPN and IL-17 levels in peripheral blood of hepatitis B patients are closely related to the hepatitis B degree.TNF-α can increase the level of OPN secreted by lymphocytes.

Objective To understand the elonal expansion and genetic diversity of Salmonella en-terica semtype Paratyphi A (SPA) and to construct a typing method to determine the epidemic clones of the isolates. Methods Antimicrobial susceptibility testing was performed with 3980 SPA isolates by the cen-trolled Kirby-Bauer disc diffusion technique on Muller-Hinton agar plates. A total of 15 SPA with nalidixie acid resistance for mutations in gyrA, gyrB, gyrC and gyrE genes within the quinolone-resistant determina-tion region (QRDR) were examined. Subtyping of 121 isolates of SPA from seven counties in Yuxi were studied using pulsed-field gel eleetrophoresis (PFGE) analysis following digestion of chromosomal DNA with restriction endanucleases Spe Ⅰ and Xba Ⅰ. PFGE patterns were analyzed by duster analysis. Results The nalidixic acid-susceptible isolates predominated in 1999 but was replaced by nalidixic acid -resistant (NAR) isolates after 2000. Amplification by PCR and sequencing of the genes with subsets of 15 NAR strains re-vealed that the resistance mechanisms had resulted from single point mutations in the gyrA gene. Spe Ⅰ and Xba Ⅰ digestion of 121 isolates gave five and four different PFGE patterns with the predominance of the Spe Ⅰ 01 and Spe Ⅰ 02 (or the Xba Ⅰ 01) epidemic patterns, respectively. Spe Ⅰ 01 and Spe Ⅰ 02 consisted of 37.2% and 57.9% of isolates, respectively, or Xba Ⅰ 01 consisted of 95.0% of isolates. Conclusion The incidence of resistance to nalidixic acid of the isolates increased during the study period. PFGE patterns Spe Ⅰ 01 and Spe Ⅰ 02 (or Xba Ⅰ 01), the main clones of the epidemics, are highly prevalent in Yuxi. PFGE with Spe Ⅰ and Xba Ⅰ is a useful technique to differentiate SPA.