Capture of cervical exfoliative cells on a glass slide coated by 3-glycidyloxypropyl trimethoxysilane and poly-L-lysine

A new modification method for glass slides was developed and applied to make ThinPrep Pap smears,in order to increase the adhesion ability of cervical exfoliative cells.3-glycidyloxypropyl trimethoxysilane(GOPS) was coated on the glass slides firstly on the slides,then poly-L-lysine(PLL) was covalently modified onto the above epoxy-terminated slides to form GOPS-PLL double decorated slides.The modified slides were characterized using X-ray photoelectron spectroscopy(XPS) and atomic force microscopy(AFM).The cell adhesion ability effect was tested and compared with traditional PLL coated slides by fixing the cervical exfoliative cells on the double adorned slides.The control test was conducted by the bare glass slides unmodified.The cell morphology of cervical exfoliative cells adhered on different slides was observed under the microscope after Papanicolaou staining.The number of cervical exfoliative cells on the unmodified slides,PLL coated slides and GOPS-PLL coated slides was 10307300,32837226 and 41197280(n=12),respectively.The data among the three different modification methods showed significant differences(one-way analysis of variance,ANOVA test,Po0.05).The cell capturing effect of the GOPS-PLL slide was the best among the three different modified slides.In addition,the GOPS-PLL slide could enhance the uniformity of the adhered cells and be widely applied to the ThinPrep system for cervical carcinoma screening to increase the accuracy rate of diagnosis.

DIAGNOSTIC VALUE OF DENSITY GRADIENT CENTRIFU-GATION FOR EXFOLIATIVE TUMOR CELLS IN MALIG-NANT PLEURAL EFFUSIONS

Objective To find out a specific method for diagnosis of malignant pleural effusions (MPEs)with higher sensitivity and practicality. Methods The diagnosis of MPEs were made using density gradient cen-trifugation(DGC) , smear cytologic examination( SCE) and pleural needle biopsy (PNB). Comparisons between these results and those of benign pleural effusions were also made. Results The positive rates of DGC,SCE and PNB for diagnosing MPEs were 94. 3% ,62. 9% and 44. 6% , respectively, and the positive rate of SCE combined with PNB for diagnosing MPEs was 73. 2%. The positive rate of the exfoliative tumor cells ( ETCs ) by DGC was much higher than that of SCE or/and PNB with no false-positive. Conclusion The ETCs isolated by DGC from the MPEs is quite specific for the diagnosis of malignant tumors with higher sensitivity and practicality in clinico-patho-logical practice.

Analysis of Human Papillomavirus Infection in Cervical Exfoliated Cells

To screen patients with early cervical lesions by analyzing the infection of high-risk Human papillomavirus (HR-HPV). Research Methods: The cervical exfoliated cell specimens and their clinical data were collected. The HPV infection types of the collected specimens were detected by fluorescence quantitative PCR, and the correlation between HPV infection and clinicopathological features was analyzed statistically. Results: 725 cases were HR-HPV positive from 2605 cases, including 15 high-risk types of HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68. Different histological types ranged from NILM to HSIL, and the positive rate of HPV showed an increasing trend with the aggravation of cervical lesions. Conclusion: The positive rate of 15 high-risk HPV types in the collected specimens was 27.8%. Patients with early cervical lesions could be screened for 15 high-risk HPV infection types.

Stem cells from human exfoliated deciduous teeth ameliorate concanavalin A-induced autoimmune hepatitis by protecting hepatocytes from apoptosis

BACKGROUND Autoimmune hepatitis is a serious autoimmune liver disease that threatens human health worldwide,which emphasizes the urgent need to identify novel treatments.Stem cells from human exfoliated deciduous teeth(SHED),which are easy to obtain in a non-invasive manner,show pronounced proliferative and immunomodulatory capacities.AIM To investigate the protective effects of SHED on concanavalin A(ConA)-induced hepatitis in mice,and to elucidate the associated regulatory mechanisms.METHODS We used a ConA-induced acute hepatitis mouse model and an in vitro co-culture system to study the protective effects of SHED on ConA-induced autoimmune hepatitis,as well as the associated underlying mechanisms.RESULTS SHED infusion could prevent aberrant histopathological liver architecture caused by ConA-induced infiltration of CD3+,CD4+,tumor necrosis-alpha+,and interferon-gamma+inflammatory cells.Alanine aminotransferase and aspartate aminotransferase were significantly elevated in hepatitis mice.SHED infusion could therefore block ConA-induced alanine aminotransferase and aspartate aminotransferase elevations.Mechanistically,ConA upregulated tumor necrosisalpha and interferon-gamma expression,which was activated by the nuclear factor-kappa B pathway to induce hepatocyte apoptosis,resulting in acute liver injury.SHED administration protected hepatocytes from ConA-induced apoptosis.CONCLUSION SHED alleviates ConA-induced acute liver injury via inhibition of hepatocyte apoptosis mediated by the nuclear factor-kappa B pathway.Our findings could provide a potential treatment strategy for hepatitis.

Application of dental stem cells in three-dimensional tissue regeneration

Dental stem cells can differentiate into different types of cells.Dental pulp stem cells,stem cells from human exfoliated deciduous teeth,periodontal ligament stem cells,stem cells from apical papilla,and dental follicle progenitor cells are five different types of dental stem cells that have been identified during different stages of tooth development.The availability of dental stem cells from discarded or removed teeth makes them promising candidates for tissue engineering.In recent years,three-dimensional(3D)tissue scaffolds have been used to reconstruct and restore different anatomical defects.With rapid advances in 3D tissue engineering,dental stem cells have been used in the regeneration of 3D engineered tissue.This review presents an overview of different types of dental stem cells used in 3D tissue regeneration,which are currently the most common type of stem cells used to treat human tissue conditions.

Isolated repeated anastomotic recurrence after sigmoidectomy

Repeated anastomotic recurrence(AR)of colonic cancer is uncommon.We report a case of a double-isolated AR after sigmoidectomy.In 2003,a 60-year-old woman underwent stapled sigmoid resection for a moderately differentiated adenocarcinoma.Further rectal bleeding occurred after six months,and colonoscopy detected an AR.Thus,an additional stapled colorectal anastomosis was performed.Ten months later,a colonoscopy detected a circumferential AR that prompted the completion of a second colorectal resection,with a doublestapled colorectal anastomosis.Twenty-four hours after surgery,a massive pulmonary embolism occurred,and the patient died within a few hours.At present,onlysix cases of repeated isolated AR have been described.Repeated segmental colorectal resections are generally associated with a favourable prognosis,with a median survival rate of 45 mo(range,13-132 mo).Repeated isolated ARs are rare,and segmental colorectal resections are generally associated with long-term diseasefree survival.

SHED-derived exosomes ameliorate hyposalivation caused by Sjögren’s syndrome via Akt/GSK-3b/Slug-mediated ZO-1 expression

Background:Sjögren’s syndrome(SS)is an autoimmune disorder characterized by sicca syndrome and/or systemic manifestations.The treatment is still challenging.This study aimed to explore the therapeutic role and mechanism of exosomes obtained from the supernatant of stem cells derived from human exfoliated deciduous teeth(SHED-exos)in sialadenitis caused by SS.Methods:SHED-exos were administered to the submandibular glands(SMGs)of 14-week-old non-obese diabetic(NOD)mice,an animal model of the clinical phase of SS,by local injection or intraductal infusion.The saliva flow rate was measured after pilocarpine intraperitoneal injection in 21-week-old NOD mice.Protein expression was examined by western blot analysis.Exosomal microRNA(miRNAs)were identified by microarray analysis.Paracellular permeability was evaluated by transepithelial electrical resistance measurement.Results:SHED-exos were injected into the SMG of NOD mice and increased saliva secretion.The injected SHED-exos were taken up by glandular epithelial cells,and further increased paracellular permeability mediated by zonula occluden-1(ZO-1).A total of 180 exosomal miRNAs were identified from SHED-exos,and Kyoto Encyclopedia of Genes and Genomes analysis suggested that the phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt)pathway might play an important role.SHED-exos treatment down-regulated phospho-Akt(p-Akt)/Akt,phospho-glycogen synthase kinase 3b(p-GSK-3b)/GSK-3b,and Slug expressions and up-regulated ZO-1 expression in SMGs and SMG-C6 cells.Both the increased ZO-1 expression and paracellular permeability induced by SHED-exos were abolished by insulin-like growth factor 1,a PI3K agonist.Slug bound to the ZO-1 promoter and suppressed its expression.For safer and more effective clinical application,SHED-exos were intraductally infused into the SMGs of NOD mice,and saliva secretion was increased and accompanied by decreased levels of p-Akt/Akt,p-GSK-3b/GSK-3b,and Slug and increased ZO-1 expression.Conclusion:Local application of SHED-exos in SMGs can ameliorate Sjögren syndrome-induced hyposalivation by increasing the paracellular permeability of glandular epithelial cells through Akt/GSK-3b/Slug pathway-mediated ZO-1 expression.

Bioinspired porous microspheres for sustained hypoxic exosomes release and vascularized bone regeneration

Exosomes derived from mesenchymal stem cells(MSCs)have demonstrated regenerative potential for cell-free bone tissue engineering,nevertheless,certain challenges,including the confined therapeutic potency of exosomes and ineffective delivery method,are still persisted.Here,we confirmed that hypoxic precondition could induce enhanced secretion of exosomes from stem cells from human exfoliated deciduous teeth(SHEDs)via comprehensive proteomics analysis,and the corresponding hypoxic exosomes(H-Exo)exhibited superior potential in promoting cellular angiogenesis and osteogenesis via the significant up-regulation in focal adhesion,VEGF signaling pathway,and thyroid hormone synthesis.Then,we developed a platform technology enabling the effective delivery of hypoxic exosomes with sustained release kinetics to irregular-shaped bone defects via injection.This platform is based on a simple adsorbing technique,where exosomes are adsorbed onto the surface of injectable porous poly(lactide-co-glycolide)(PLGA)microspheres with bioinspired polydopamine(PDA)coating(PMS-PDA microspheres).The PMS-PDA microspheres could effectively adsorb exosomes,show sustained release of H-Exo for 21 days with high bioactivity,and induce vascularized bone regeneration in 5-mm rat calvarial defect.These findings indicate that the hypoxic precondition and PMS-PDA porous microsphere-based exosome delivery are efficient in inducing tissue regeneration,hence facilitating the clinical translation of exosome-based therapy.