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β-Thalassemia caused by abnormal coding of the β-globin gene is the most common hemoglobinopathy in many Asian countries. The in-depth study of the molecular basis and epigenetic mechanism of globin gene expression is the key to explore a new treatment for thalassemia. In this study, FAIRE (formaldehyde-assisted isolation of regulatory elements), 3C (chromosome conformation capture) and ChIP (Chromatin Immunoprecipitation) were used to investigate the three-dimensional interaction network of β-globin family gene loci and the molecular mechanism of functional regulation of gene expression during rapamycin-induced chromatin remodeling in CD4+ T cells. The results showed that the opening degree of globin gene chromatin, the interaction frequency between the gene promoter region and the regulatory element LCR (Locus control regions), and the enrichment efficiency of CTCF (CCCTC-binding factor) in the gene promoter region changed differently during the change of rapamycin treatment concentration from low to high, which led to the same change trend of the gene expression pattern. At the 10 nmol/ L concentration, chromatin accessibility and gene expression decreased (P < 0. 05). At 20 nmol/ L and 50 nmol/ L concentrations, chromatin accessibility increased and gene expression was up-regulated (P < 0. 05). In this study, the molecular mechanism of gene expression regulation of the β-globin family was expounded through this dynamic change process. Our work provides a theoretical and clinical practice basis for clinical precision treatment.
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OBJECTIVE@#To investigate the correlation of inducible co-stimulatory molecules (ICOS) with mesenteric vascular endothelial- mesenchymal transition (EndMT) and sclerosis in spontaneously hypertensive rats (SHR).@*METHODS@#Twenty 4-week-old WKY rats and 20 SHRs of the same strain were both randomly divided into 4 groups for observation at 4, 6, 10 and 30 weeks of age. ICOS expression frequency in rat spleen CD4+T cells was analyzed using flow cytometry, and the expressions of ICOS, VE-cad, α-SMA and Col3 mRNA in rat mesentery were detected by RT-PCR. The distributions of ICOS, IL-17A and TGF-β in rat mesentery were detected by immunohistochemistry. The levels of IL-17A and TGF-β in rat plasma were measured using ELISA. The morphological changes of rat mesenteric vessels were observed with Masson staining. Spearman or Pearson correlation analyses were used to evaluate the correlation between ICOS expression and the expressions of the markers of vascular EndMT and sclerosis.@*RESULTS@#Compared with the control WKY rats, the SHRs began to show significantly increased systolic blood pressure and ICOS expression frequency on CD4+T cells at 6 weeks of age (P < 0.05). In the SHRs, the mRNA and protein expressions of ICOS, α-SMA, Col3, IL-17A and TGF-β in the mesentery were significantly higher than those in control group (P < 0.05), while the mRNA and protein expressions of VE-cad started to reduce significantly at 10 weeks of age (P < 0.05). The plasma levels of IL-17A and TGF-β were significantly increased in SHRs since 6 weeks of age (P < 0.05) with progressive worsening of mesenteric vascular sclerosis (P < 0.05). ICOS mRNA and protein expression levels in the mesenteric tissues of SHRs began to show positive correlations with α-SMA and Col3 expression levels and the severity of vascular sclerosis at 6 weeks of age (P < 0.05) and a negative correlation with VE-cad expression level at 10 weeks (P < 0.05).@*CONCLUSION@#ICOS play an important pathogenic role in EndMT and sclerosis of mesenteric vessels in essential hypertension by mediating related immune responses.
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Rats , Animals , Rats, Inbred SHR , Rats, Inbred WKY , Hypertension , Interleukin-17 , Sclerosis/pathology , Transforming Growth Factor beta , Mesentery/pathology , RNA, Messenger/metabolism , Blood PressureABSTRACT
OBJECTIVE@#To explore the role of interleukin-17A (IL-17A) in renal epithelial- mesenchymal transition (EMT) in essential hypertensive nephropathy.@*METHODS@#Four-week-old spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats (control group) were both randomized into 4 groups (n=5) for observation at 4, 6, 10 and 30 weeks of age. Blood pressure of the rats was monitored using a noninvasive tail artery blood pressure measurement instrument. The percentage of Th17 cells in the splenocytes was analyzed using flow cytometry. The mRNA and protein expression levels of IL-17A, iNOS, Arg-1, E-cadherin, and α-SMA in the kidneys of the rats were detected using RT-PCR and immunohistochemical staining, respectively, and plasma levels of IL-17A were regularly detected using ELISA.@*RESULTS@#At the age of 6 weeks, the SHRs began to show significantly higher blood pressure with greater Th17 cell percentage in the splenocytes and high renal expression and plasma level of IL-17A than WKY rats (P < 0.05 or P < 0.01). At 30 weeks, renal expression of E-cadherin mRNA and protein was significantly lower and the expression of Arg-1 mRNA and protein was significantly higher in SHR than in WKY rats (P < 0.01). Compared with the WKY rats, the SHRs showed significantly higher mRNA and protein expressions of iNOS at 6 and 10 weeks (P < 0.05 or 0.01) and higher α-SMA mRNA and protein expressions since 10 weeks of age (P < 0.05 or 0.01). In SHRs older than 10 weeks, renal IL-17A mRNA and protein expression levels were negatively correlated with those of E-cadherin (r=-0.731, P < 0.05; r=-0.827, P < 0.01) and positively correlated with those of α-SMA (r=0.658, P < 0.05; r=0.968, P < 0.01).@*CONCLUSION@#IL-17A is closely correlated with the progression of renal EMT in SHR and plays its role possibly by mediating M1/M2 polarization of renal infiltrating macrophages.
Subject(s)
Animals , Rats , Blood Pressure , Cadherins/metabolism , Epithelial-Mesenchymal Transition , Hypertension , Interleukin-17/metabolism , Kidney , RNA, Messenger/metabolism , Rats, Inbred SHR , Rats, Inbred WKYABSTRACT
Objective:To explore the soft tissue composition underlying upper eyelid puffiness and the experience in treating it with tailor-made procedure during upper blepharoplasty.Methods:In a retrospective study from January 2017 to December 2018, 165 patients with upper eyelid puffiness underwent full-incision upper blepharoplasty. During the procedure, the anterior wall of orbital septum was exposed, and the volume of the retro-orbicularis oculus fat (ROOF) and preaponeurotic fat was observed. The orbital septum would be opened if lacrimal gland prolapse was suspected. The puffy upper eyelid would be targeted with tailor-made procedure such as partial ROOF resection, removal or transposition of the preaponeurotic fat, as well as lacrimal gland refixation.Results:All 165 cases were healed by first intention. No relapse was found after a 6 to 24 months follow-up. Persistent mild pain and local skin tightness epiphora chemosis were presented in isolated cases with self-resolution within a short time. There were no severe adverse reaction or complications occurred during the operation and follow-up.Conclusions:The tailor-made procedure during upper blepharoplasty targets the soft tissue anatomy underlying upper eyelid puffiness. It is safe and effective in maintaining a natural look of upper eyelids while retaining the appropriate amount of adipose tissue in the upper eyelid.
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Objective@#To evaluate the clinical outcomes of a series of patients who have undergone reconstruction of craniofacial defects after resection of intracranial tumors or craniofacial trauma with free composite anterolateral thigh flaps.@*Methods@#Retrospective analyses the clinical cases from September 2007 to September 2016. Data included flap survival rate, complication, satisfaction survey was reviewed to evaluate the efficacy and safety of this surgical strategy.@*Results@#Totally 10 free anterolateral thigh flaps including 3 cases of fasciocutaneous flaps, 2 case of adipofascial flaps, 4 cases of myocutaneous flaps, 1 case of chimeric flap, were adopted to reconstruct craniofacial defects. Follow-up ranged from 3 to 17 months (average, 12 months). All flaps were transferred successfully. There were no cranial spinal fluid(CSF) leaks, intracranial infections or donor site complications. All patients were satisfied.@*Conclusions@#Because of its abundance of tissue, matched vessels to recipient site, versatility of muscular flaps to fill irregularly intracranial defects, reliable blood supply, feasibility of simultaneous fascia lata harvesting, free composite anterolateral thigh flap is the reconstructive method of choice for craniofacial defects reconstruction after resection of intracranial tumors or craniofacial trauma. The use of ALT flap was reliable in the decrease of CSF leak and infection rate and dependable according to long time follow-up.
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Objective To investigate clinical characteristics and treatment of upper eyelid lipogranuloma cases that developed after facial autologous fat injection.Methods This retrospective study involved 8 patients who presented with upper eyelids symptoms (final diagnosis of lipogranulo ma) and had history of facial autologous fat injection.The collected data included information on patient sex,age,clinical presentation,laboratory findings,treatment and outcome.Results All 8 cases were managed by exploration and surgical removal through double eyelid surgery.All patients showed complete resolution over 1-12 months follow up period.Conclusions Lipogranuloma can develop in the upper eyelid after autologous fat injection into the face.and surgical removal can yield satisfactory outcomes in cases of repeated failure of conservative therapies.
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Objective To lay a foundation for the clinical application and tissue engineering research of hypertrophic scar (HS)-derived DMSCs by comparing the biological characteristics of dermis mesenchymal stem cells (DMSCs) from human maturing-phase HS and normal skin.Methods Twenty maturing-phase HS specimens (scar group) and 20 normal skin specimens (control group) were selected to extract and sort DMSCs by two-step enzyme digestion.When cells in both groups were subcultured to 3rd generation,cell morphology and growth curve were observed; expressions of cell surface proteins CD29,CD49 and vimentin were tested by immunocytochemistry; cells with positively expressed surface proteins CD34,CD73,CD90,and CD105 were examined by flow cytometry; expressions of genes Oct4 and Nanog were tested by RT-PCR; cell potential to differentiate into lipoblasts,osteoblasts,and chondroblasts was assayed in inductive medium.Results DMSCs in both groups showed similar shape and growth curve.Cell markers CD29,CD49 and vimentin expressed positively.Of scar and control groups,expressions of CD34,CD73,CD90,and CD105 were (0.60±0.03)％ vs (0.61 ±0.02)％,(98.90±0.80)％vs (99.00±0.70)％,(98.30±0.30)％vs (98.20±0.40)％,and (93.10± 0.40) ％ vs (93.00 ± 0.20) ％ respectively (P ＞ 0.05) ; expressions of genes Oct4 and Nanog were 0.506±0.024 vs0.512±0.024 and 0.496 ±0.018 vs 0.494 ±0.023 (P＞0.05).Both types of DMSCs were able to differentiate in vitro into lipoblasts,osteoblasts,and chondroblasts in invitro conductive medium.Conclusion DMSCs exist in maturing-phase HS and present biomechanical characteristics basically similar with those of normal human skin.
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<p><b>OBJECTIVE</b>To investigate the effect of epidermal stem cells from human hypertrophic scar (HS-ESCs) on the skin wound healing in nude mice.</p><p><b>METHODS</b>40 mice were randomly divided into two groups as experimental group (n = 20) and control group (n = 20). Wounds, 1 cm in diameters, were made on every mouse back. The wounds were treated with HS-ESCs and erythromycin ointment in experimental group, or only with erythromycin ointment in control group. The wound healing was observed during the following 14 days. The expression of collagen-I, collagen-III, epidermal growth factor (EGF), fibroblast growth factor (FGF2) , transforming growth factor (TGFbeta1, and TGFbeta2) were studied.</p><p><b>RESULTS</b>The wound healing time in the experimental group was (20.8 +/- 0.84) d, which was (25.6 +/- 0.89) d in the control group. HE staining revealed that the extent of vascularization in the experimental group was 11.60 +/- 0.55, while it was 8.04 +/- 0.33 in the control group. Immunochemistry analysis showed the expression of collagen-I, collagen-III, EGF, FGF2, TGFbeta1, and TGFbeta2 in the experimental group were significantly higher, compared with those in control group (P < 0.05).</p><p><b>CONCLUSION</b>HS-ESCs may promote wound healing through enhancement of the vascularization of the wound tissue and the expression of growth factors.</p>
Subject(s)
Animals , Female , Humans , Male , Mice , Cicatrix, Hypertrophic , Pathology , Epidermis , Cell Biology , Mice, Nude , Skin , Wounds and Injuries , Stem Cell Transplantation , Stem Cells , Wound HealingABSTRACT
<p><b>OBJECTIVE</b>To explore the presence of bacterial biofilms (BF) in patients with CRS and the effect of BF on clinical symptoms and postoperative outcomes.</p><p><b>METHODS</b>Seventy-two patients with chronic sinusitis were enrolled in this study. The control group included 15 patients with deviation of the nasal septum and 10 patients had a fracture of the nasal bone. Mucosa of the uncinate process or ethmoid near the ostium of the maxillary sinus was obtained during endoscopic sinus surgery. The specimens were subjected to scanning electron microscopy. Patients were followed for 1 year and observed by the Lund-Kennedy endoscopy, and the Haikou standard classification (ESS-1997). Statistical analysis was performed by t-test or chi-square test.</p><p><b>RESULTS</b>Three patients were lost to follow-up. The scanning electron microscopy analysis showed bacterial biofilms in 49 of the 69 patients with chronic sinusitis. A marked destruction of the epithelium and cilia was observed in samples positive for bacterial biofilms. No bacterial biofilms were detected in the control group, and scanning electron microscopy showed normal epithelium and cilia in those specimens. There was no significant difference in gender, classification or duration of disease between the BF(-) and BF(+) groups. At six months and one year postoperative, the Lund-Kennedy endoscopy scores for CRS patients with BF (4.78 +/- 1.67; 4.55 +/- 1.61) were significantly higher than those without BF (3.65 +/- 1.39; 3.65 +/- 1.18) (t = -2.654, P < 0.01; t = -2.264, P < 0.05). Based on the Haikou standard classification, there was a significantly difference between patients with BF and those without BF (chi2 = 18.014, 22.063, P < 0.001, respectively).</p><p><b>CONCLUSIONS</b>Different life stages of bacterial biofilms were demonstrated to be present in CRS. Gender, classification or duration of disease did not affect the presence of bacterial biofilms in patients with CRS. There is a correlation between bacterial biofilms and an unfavorable outcome in patients with CRS after ESS.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Biofilms , Case-Control Studies , Chronic Disease , Follow-Up Studies , Microscopy, Electron, Scanning , Nasal Mucosa , Microbiology , Nasal Polyps , Microbiology , Rhinitis , Microbiology , Sinusitis , MicrobiologyABSTRACT
<p><b>OBJECTIVE</b>To study the surgical treatment of recurrent laryngeal nerve (RLN) injury caused by thyroid operation.</p><p><b>METHODS</b>From 1970 to 2001, 50 patients with RLN injury were caused by thyroid operation. The causes, location, type, operative procedures and follow-up were retrospectively analyzed.</p><p><b>RESULTS</b>Unilateral RLN injury occurred in 46 cases and bilateral nerve injury in 4 cases. The RLN injuries were located within 2cm below the point of RLN entering to throat in 45 nerves (83.3%), other places in 6 nerves (11.3%), and unknown location in 3 nerves (5.4%). Transection of the nerve was found in 19 nerves (36.5%), suture or scare pressing the nerve in 35 nerves (64.8%). All the injured nerves were repaired surgically. Meanwhile all 4 patients with bilateral RLN injuries underwent tracheotomy. Of the 50 cases, 44 cases (88.0%) were followed up for more than 1.5 years. Among the 44 followed-up patients, phonation was restored to normal or obvious improvement in 42 cases (95.5%), and improvement in 2 (4.5%). Of the 35 patients with 39 nerves underwent indirect or direct laryngoscopy, the affected vocal cord movement entirely recovered in 21 cords (53.8%), partially recovered in 7 cords (17.9%), uncovered in 11 cords (28.3%). There was no relation between the recovery of phonation or vocal cord movement with the timing or the procedure of repairing operation.</p><p><b>CONCLUSIONS</b>The location of most RLN injuries caused by thyroid surgery are just below the point of RLN entering to throat, and most are mechanical injury, and need operation to resolve the cause. Once the RLN injury is made, an operation should be performed as early as possible.</p>