Alternative splicing and intron retention: Their profiles and roles in cutaneous fibrosis of systemic sclerosis.

BACKGROUND: Alternative splicing (AS) and intron retention (IR) implicated in multiple pathophysiological processes, have rarely been reported in systemic sclerosis (SSc). METHODS: We integrated bulk RNA-seq and 4D label-free mass spectrometry to perform a multi-omics analysis of AS and IR in SSc skin tissue and fibroblasts. RMATS and iREAD were used to identify AS and IR, which were validated by real-time PCR. Spearman correlation and the LASSO method were employed to assess correlations among clinical features, introns, splicing factors (regulators of AS) and proteins. FINDINGS: AS profiles showed distinct alterations in SSc skin tissue, with the most pronounced changes occurring in IR. AS and IR were associated with total modified Rodnan skin score (mRSS) and local skin score. Upon TGF-ß stimulation, fibroblasts exhibited significant alterations in IR profiles, affecting genes related to fibroblast proliferation and collagen fibril organization. A comprehensive integrated analysis of introns, exons, and proteome profiles revealed that IR exerted a negative impact on protein expression, with certain changes being under intronic control. RT-PCR confirmed the presence of intron and exon-derived sequences of CTTN, OGA, MED16 and PHYKPL. Additionally, notable changes were observed in the regulatory network of splicing factors in SSc skin tissues. These factors are also involved in fibrosis pathways and correlated with clinical features. CONCLUSION: Totally, abnormal AS, IR profiles and splicing factors were identified in SSc, altered IRs and splicing factors participated in fibrosis-related pathways. IR exerted a negative impact on protein expression in TGF-ß-stimulated fibroblasts. Clarification of the IR mechanisms will provide new insights into the pathophysiology of SSc.

Crocin suppresses breast cancer cell proliferation by down-regulating tumor promoter miR-122-5p and up-regulating tumor suppressors FOXP2 and SPRY2.

Crocin has been reported to have antitumor activity in several tumors including breast cancer. Nevertheless, the mechanism of action of crocin on breast cancer remains unclear. The cytotoxicity of crocin was evaluated by CCK-8 assay. Cell proliferation was assessed using EdU incorporation assay and western blot analysis. Breast cancer-related genes were extracted from GEPIA. miR-122-5p targets were predicted using Targetscan, starbase, and miRDB softwares. Luciferase reporter assay was employed to confirm whether miR-122-5p targeted sprouty2 (SPRY2) and forkhead box P2 (FOXP2). Results showed that crocin exhibited cytotoxicity and suppressed the proliferation in breast cancer cells. miR-122-5p was upregulated in breast cancer tissues and cells. Crocin suppressed miR-122-5p to block the proliferation of breast cancer cells. Seven targets of miR-122-5p were identified in breast cancer. SPRY2 and FOXP2 were selected for further experiments due to their involvement in breast cancer. miR-122-5p targeted SPRY2 and FOXP2 to inhibit their expression. miR-122-5p knockdown restrained breast cancer cell proliferation by targeting SPRY2 and FOXP2. Additionally, crocin increased SPRY2 and FOXP2 expression by inhibiting miR-122-5p expression. Together, our results suggested that crocin inhibited proliferation of breast cancer cells through decreasing miR-122-5p expression and the subsequent increase of SPRY2 and FOXP2 expression.

ABCA8 inhibits breast cancer cell proliferation by regulating the AMP activated protein kinase/mammalian target of rapamycin signaling pathway.

ATP-binding cassette (ABC) subfamily A member 8 (ABCA8) has been reported to play a vital role in cancer development. Our study aimed to explore the role and the molecular mechanism of ABCA8 in breast cancer (BC) progression. GSE65194, GSE15852, and GSE45827 datasets were used to identify differentially expressed genes (DEGs) in BC. The diagnosis and prognosis value were determined using ROC curve analysis and Kaplan-Meier plotter, respectively. The relationship between ABCA8 expression and clinicopathological features in BC was analyzed by TCGA. Co-expressed genes of ABCA8 in BC were screened out through GEPIA and subjected to KEGG pathway enrichment analysis. Cell proliferation was evaluated by CCK-8 and EdU incorporation assays. Proliferating cell nuclear antigen (PCNA) expression and the changes of the AMP activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway were measured by western blot analysis. Totally 4 overlapping DEGs were identified and all reduced in BC samples. ABCA8 with high diagnostic and prognostic values was selected for further exploration. Low ABCA8 expression was correlated with clinicopathological features in BC patients. ABCA8 overexpression inhibited BC cell proliferation. The top 20 co-expressed genes of ABCA8 were identified by GEPIA and significantly enriched in AMPK signaling pathway. Inhibition of AMPK/mTOR pathway reversed the suppressive effect of ABCA8 on BC cell growth. These results suggested that ABCA8 overexpression repressed BC cell proliferation through regulating the AMPK/mTOR signaling pathway.

The cytotoxicity of karanjin toward breast cancer cells is involved in the PI3K/Akt signaling pathway.

Karanjin is a bioactive furanoflavonoid with various pharmacological activities including anticancer activities. However, the effect and the related mechanism of karanjin in breast cancer (BC) have not been revealed. The potential targets of karanjin and BC were predicted using SwissTargetPrediction and GeneCards databases, respectively. The overlapping targets between karanjin and BC were identified using the Venn diagram. DAVID database was used for the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment pathway analysis. Cell viability, proliferation, and apoptosis were examined by MTT (3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-tetrazolium bromide), EdU (5-ethynyl-2'-deoxyuridine) incorporation, and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP digoxigenin nick-end labeling) assays, respectively. The protein levels were measured by western blot analysis. We screened out 28 overlapping targets between karanjin and BC. KEGG analysis showed that the targets of karanjin in BC were associated with the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway. Karanjin inhibited cell viability and impeded the proliferative ability of BC cells. Moreover, karanjin treatment induced apoptosis in BC cells. Additionally, karanjin treatment blocked the PI3K/Akt signaling pathway and activation of the PI3K/Akt pathway reversed the antitumor effect of karanjin on BC cells. In conclusion, karanjin exerted antitumor activity in BC cells by regulating the PI3K/Akt signaling pathway.

Postoperative drain after transoral endoscopic thyroidectomy vestibular approach (TOETVA) with single incision.

OBJECTIVE: To find the optimal size of a drain for the reliable drainage and the best cosmetic result in TOETVA. To explore the normal drainage flow rate after TOETVA. METHODS: A prospective randomized controlled trial was performed in a single center from December 2016 to December 2018. One hundred and fifty-three (153) patients had TOETVA with a single incision and were randomly divided into two groups. Self-made drainage tubes with a small diameter (outer diameter 2.0 mm, inner diameter 1.0 mm) were used in 80 patients (experimental group). No. 8 tubes were used in 73 patients (control group). The clinical characteristics and results between both groups were compared by t test or chi-square test, and the results of normal drainage flow rate were calculated. RESULTS: The experimental group had a longer intraoperative tube-inserting time, compared with the control group (9.5 ± 2.5 min vs. 5.6 ± 1.4 min, p = 0.001), a smaller scar six months after the operation (1.8 ± 2.3 mm vs. 3.1 ± 2.6 mm, p = 0.002), and a lower Vancouver Scar Scale score at both one month (3.20 ± 1.44 vs. 4.19 ± 1.92, p = 0.001) and six months after the operation(1.43 ± 1.84 vs. 2.40 ± 2.37, p = 0.006). The drainage volume, pain score on the first day, postoperative complications (tube blockage, air leakage, subcutaneous hydrops, hematoma, regional infection), and the extubation time were not significantly different. The average drainage of 148 patients without postoperative complications was 78.3 ± 10.9 ml. The cumulative drainage within 8 h, and 32 h after the operation accounted for 53.2% and 91.9% of the total drainage, respectively. The residual drainage at 32 h was estimated to be 6.5 ± 2.9 ml (P95 = 11.0 ml). A linear regression equation between total volume (Vt) and the size of resected tissue (S) was established: Vt = 1.625 S + 56.604 (p = 0.0001). CONCLUSION: In TOETVA, a small drain can provide a good cosmetic appearance and reliable drainage. The main exudation period of the wound is within 8 h after the operation. If a residual volume of less than 11 ml is considered to be self-absorbable, the shortest safe extubation point for 95% of patients should be 32 h after the operation.

The clinical application of mental nerve dissection in transoral endoscopic thyroidectomy via an oral vestibular approach.

BACKGROUND: Transoral endoscopic thyroidectomy vestibular approach is the natural orifice surgery to avoid surgical scars. However, mental nerve injury is a characteristic complication. Herein, we report the development of a novel method to dissect the mental nerve proactively during surgery to minimize the morbidity from mental nerve injury. METHODS: In this study, a total of 105 patients from June 2016 to February 2018 were categorized as the mental nerve dissection group (MND) or not mental nerve dissection group (NMND). We analyzed the demographics, operative data, hospital stay, pathologic results, and postoperative complications between the two groups. RESULTS: There were no significant differences with respect to age, gender, tumor size, extent of surgery, the amount of bleeding, or postoperative hospitalization between groups. The specimen removal time was shorter in the MND group. The average operation time in both groups was similar, but the operation time for a hemithyroidectomy with CND in the MND group was shorter than in the NMND group. The VAS pain scores and complication rates that included transient hypocalcemia, seroma, subcutaneous emphysema, transient, and permanent recurrent laryngeal nerve palsy did not differ significantly between groups. In terms of mental nerve injury, the morbidity rate in the MND group was lower than in the NMND group. CONCLUSIONS: The modified endoscopic thyroidectomy involving dissection of the mental nerve via the oral vestibular approach is safe and feasible. It is beneficial to protect the mental nerve and for specimen removal which is worth clinical promotion.

The expression and proangiogenic effect of nucleolin during the recovery of heat-denatured HUVECs.

BACKGROUND: The present study aims to examine the expression patterns and roles of nucleolin during the recovery of heat-denatured human umbilical vein endothelial cells (HUVECs). METHODS: Deep partial thickness burn model in Sprague-Dawley rats and the heat denatured cell model (52°C, 35s) were used. The expression of nucleolin was measured using Western blot analysis and real-time PCR. Angiogenesis was assessed using in vitro parameters including endothelial cell proliferation, transwell migration assay, and scratched wound healing. Gene transfection and RNA interference approaches were employed to investigate the roles of nucleolin. RESULTS: Nucleolin mRNA and protein expression showed a time-dependent increase during the recovery of heat-denatured dermis and HUVECs. Heat-denaturation time-dependently promoted cell growth, adhesion, migration, scratched wound healing and formation of tube-like structures in HUVECs. These effects of heat denaturation on endothelial wound healing and formation of tube-like structures were prevented by knockdown of nucleolin, whereas over-expression of nucleolin increased cell growth, migration, and formation of tube-like structures in cultured HUVEC endothelial cells. In addition, we found that the expression of vascular endothelial growth factor (VEGF) increased during the recovery of heat-denatured dermis and HUVECs, and nucleolin up-regulated VEGF in HUVECs. CONCLUSIONS: The present study reveals that the expression of nucleolin is up-regulated, and plays a pro-angiogenic role during the recovery of heat-denatured dermis and its mechanism is probably dependent on production of VEGF. GENERAL SIGNIFICANCE: We find a novel and important pro-angiogenic role of nucleolin during the recovery of heat-denatured dermis.

Cancer-associated fibroblasts-derived lncRNA signature as a putative biomarker in breast cancer.

Long noncoding RNAs (lncRNAs) have been reported to play a key role in regulating tumor microenvironment and immunity. Cancer-associated fibroblasts (CAFs) are abundant in many tumors. However, the functional and clinical significance of lncRNAs specifically expressed in CAFs has not been fully elucidated. In this study, we identified a list of 95 CAF-specific lncRNAs (FibLnc), including HHLA3, TP53TG1, ST7-AS1, LINC00536, ZNF503-AS1, MIR22HG, and MAPT-AS1, based on immune cell transcriptome expression profiling data. Based on the Cancer Genome Atlas and Gene Expression Omnibus datasets, we found that the FibLnc score predicted differences in overall patient survival and performed well in multiple datasets. FibLnc score was associated with the clinical stage of patients with breast cancer but did not significantly correlate with the PAM50 classification. Functional analysis showed that FibLnc was positively correlated with signaling pathways associated with malignant tumor progression. In addition, FibLnc was positively correlated with tumor mutational load and could predict immunotherapy response in patients with breast cancer receiving anti-PD-1 or anti-CTLA4 therapy. Our proposed FibLnc score was able to reflect the status of the immune environment and immunotherapeutic response in breast cancer, which could help explore potential therapeutic decisions and regulatory mechanisms of CAF-specific lncRNAs.

Reconstruction of chronic radiation-induced ulcers in the chest wall using free and pedicle flaps.

Background and purpose: Resection of radiation-induced ulcers often causes full-thickness defects of the chest wall. We retrospectively reviewed and evaluated 17 patients to explore a method of chest wall reconstruction. Materials and methods: A total of 17 breast cancer patients with radiation-induced ulcers were included. Various type of prostheses and flaps were used, results of clinic were evaluated. Results: Sixteen patients had full-thickness defects and one patient had only a soft tissue defect and underwent reconstruction with a pedicle latissimus dorsi (LD) myocutaneous flap. Among all 16 full-thickness defect cases, 15 patients underwent bony thoracic reconstruction using polymesh/3D-printed titanium plates or methyl methacrylate. For soft tissue reconstruction, 13 patients reconstruction using a free deep inferior epigastric perforator (DIEP) flap in combination with a contralateral transverse rectus abdominis myocutaneous (TRAM) flap, and 2 underwent pure free DIEP flap reconstruction. Among all the patients 15 healed with no complications, and 2 patients had delayed healing on the edges of the flaps. Conclusions: Distant pedicle or free flap can used for soft tissue defect coverage, for those severe patients with full-thickness defects and used prostheses, free deep inferior epigastric perforator flap in combination with a contralateral transverse rectus abdominis myocutaneous flap (TRAM + DIEP) would be an applicable choice.

lncRNA­MIAT facilitates the differentiation of adipose­derived mesenchymal stem cells into lymphatic endothelial cells via the miR­495/Prox1 axis.

The development of novel treatments for lymphedema is hindered by the poorly understood pathophysiology of the disease. To improve the therapeutic success of treating the disease, the present study aimed to investigate the effects and mechanism of long non­coding RNA myocardial infarction­associated transcript (MIAT) in terms of the differentiation of adipose­derived mesenchymal stem cells (ADMSCs) into lymphatic endothelial cells (LECs). The expression levels of (MIAT), microRNA (miR)­495 and Prospero­related homeobox 1 (Prox1) were measured by reverse transcription­quantitative PCR. The protein expression levels of Prox1, lymphatic vessel endothelial hyaluronan receptor 1 (LYVE­1), vascular endothelial growth factor receptor­3 (VEGFR­3) and podoplanin (PDPL) were detected by western blotting and immunofluorescence. A dual­luciferase reporter assay was also used to detect the interaction between MIAT, miR­495 and Prox1. In addition, migration and tube­formation capabilities were measured by Transwell assay and tube­formation assay, respectively. The results obtained demonstrated that VEGF­C156S (recombinant VEGF­C in which Cys156 was replaced by Ser residue) treatment could efficiently induce the differentiation of ADMSCs into LECs. MIAT expression was upregulated and miR­495 was downregulated during differentiation. Mechanistically, MIAT upregulated Prox1 expression possibly by acting as a molecular sponge for miR­495. Functional analyses indicated that the expression levels of Prox1, LYVE­1, VEGFR­3 and PDPL, and the migration and tube­formation capabilities of ADMSCs induced by VEGF­C156S, were significantly inhibited by silencing MIAT and overexpressing miR­495. Moreover, miR­495 inhibition and Prox1 overexpression reversed the effects of MIAT downregulation and miR­495 upregulation, respectively, on the differentiation of ADMSCs into LECs. Taken together, these results suggested that MIAT may be involved in the differentiation of ADMSCs into LECs, and that the MIAT/miR­495/Prox1 axis may be a novel regulatory mechanism and therapeutic target for the treatment of lymphedema.

Transoral endoscopic thyroidectomy vestibular approach in a patient with Class III goitre (operative steps and video).

Transoral endoscopic thyroidectomy vestibular approach (TOETVA) has recently become a hot research topic due to the advantage of leaving no scar, but, according to most centers, its indication is restricted to the size of thyroid gland. Here we report a case of a female patient with Class III goitre who successfully underwent TOETVA (video attached). A 53-year-old woman with a previous history of chest keloidosis presented with a history of neck swelling for 3 years and was diagnosed as Hashimoto's thyroiditis with no nodules. The patient insisted that she undergo a TOETVA procedure in our hospital. Compared to the traditional TOETVA, several techniques were applied in this operation to ensure the resection and removal of the thyroid gland: with the dissection of the mental nerve and using the lateral approach to the thyroid gland. The total volume of thyroid gland was 205 mL. The operating time was 195 min. No complications were incurred. The numbness of the lip and chin was measured by the "two-points discrimination" method with several aspects (touch, pain, temperature) at different times to evaluate mental nerve injury. She felt the numbness during the first operative day but it was almost completely relieved in the third postoperative month. To our knowledge, this is the largest thyroid gland reported removed with TOETVA procedure. By carefully designing the operative steps, it is feasible to use the transoral approach in patients who have a benign thyroid disease with a Class III goitre if the patient strongly desires that operation.

The byproduct-organic acids strengthened pretreatment of cassava straw: Optimization and kinetic study.

The subcritical liquid hot water (SLHW) pretreatment could be strengthened by its byproduct-organic acids, such as acetic acid (AA), lactic acid (LA) and formic acid (FA). The effects of these three acids on the pretreatment were investigated by the yield of fermentable sugars. The results showed that the addition of acids could effectively catalyze the hydrolysis of hemicellulose to C5 sugars and contribute to the subsequent enzymatic hydrolysis of cellulose. It was found that all three organic acids promote xylose production, and the copresence of AA + LA could limit the content of the fermentation inhibitor. The optimum proportion of three organic acids were 0.33 wt%AA + 0.45 wt%LA + 0.20 wt%FA, and the yield of C5 sugars after pretreatment and C6 sugar after enzymatic hydrolysis were 89.06% and 78.56%, respectively. The kinetic studies proved that byproduct-organic acids could promote xylose production and inhibit its further degradation and explained that xylose would accumulate at lower temperatures.

The lateral thoracic vessels: A novel recipient site for breast reconstruction with DIEP flap.

BACKGROUND: Adequate selection of recipient vessel to minimize recipient site morbidity is essential for a successful breast reconstruction. The authors explored the clinical use of the lateral thoracic vessels as recipient vessels in the deep inferior epigastric artery perforator (DIEP) flap breast reconstruction. METHODS: This study included 63 patients with breast cancer. The age of patients ranged from 26 to 67 years (43.23 ± 8.63). All patients received an immediate DIEP flap breast reconstruction using the lateral thoracic vessels as recipient vessels in the authors' department from 2014 May to 2018 September. The flap blood supply, complications, and outcomes were assessed. RESULTS: The patients were followed up for 6 to 29 months with satisfactory results. All the flaps had stable blood supply except for two cases, which had an event of partial skin necrosis and were repaired by debridement. CONCLUSIONS: Lateral thoracic vessels are not only easy to explore but also have reliable blood supply, which present a safe and reliable choice for immediate DIEP flap breast reconstruction.

Kinetic studies of the strengthening effect on liquid hot water pretreatments by organic acids.

The liquid hot water (LHW) pretreatments would be accelerated by the organic acids produced from the process. In the study, the organic acids included not only acetic acid but also lactic acid during LHW hydrolysis of reeds, at 180-220°C and for 15-135min. The lactic acid was presumably produced from xylose degradation in the pretreatment process. The different organic acids, such as acetic acid, lactic acid and acetic-lactic acids, were used to strengthen the LHW pretreatments for increasing xylose production. Moreover, the work presented kinetic models of xylose and hemicellulose at different conditions, considering the generation of lactic acid. The experimental and kinetic results both indicated that acetic-lactic acids had synergistic catalytic effect on the reaction, which could not only inhibit the degradation of xylose, but also promote the hydrolysis of hemicellulose. Besides, the highest concentration of xylose of 7.323g/L was obtained at 200°C, for 45min and with 1wt% acetic-lactic acids.

Modified transoral endoscopic thyroid surgery for treatment of thyroid cancer: operative steps and video.

In this video we describe a kind of modified transoral endoscopic thyroid surgery involving meticulous dissection of mental nerve. Inclusion criteria are: the diameter of benign tumors such as thyroid cyst, nodular goiter were limited less than 50 mm; the malignant thyroid tumors including follicular and papillary microcarcinoma were defined as a papillary carcinoma <2 cm in diameter and endoscopic surgery required for the patient. A 6 cm arc-shaped incision was designed at oral vestibule. The branches of mental nerves at both sides were identified and exposed carefully. A 10 mm trocar was placed at the midpoint of the vestibule. Two 5 mm trocars were separately inserted into the vestibule at lateral or medial of the medial branches of the mental nerve. Thyroidectomy and central lymph node dissection was done fully endoscopically using conventional endoscopic instruments.

Reconstruction of the Lower Abdominal Region Using Bilateral Pedicled Anterolateral Thigh Flaps Combined With Poly-Surgical Mesh: A Case Report.

The en-bloc resection of neoplasms on the abdominal wall often causes extensive defects that are difficult to manage. The anterolateral thigh (ALT) flap is a widely used flap in reconstructive surgery of defects. In this article, we present a case using bilateral pedicle anterolateral thigh flaps combined with a surgical polymesh to repair a large defect (22 cm × 18 cm) caused by dissection of a recurrent fibromatosis with good functional and aesthetic effects. There were no obvious morbidities or complications during a 6-month follow-up period.We conclude that the bilateral pedicle anterolateral thigh flap is a good choice for reconstruction of large lower abdominal wall defects. It can afford sufficient soft tissue coverage without obvious donor site morbidity.

Nucleolin involved in myocardial ischaemic preconditioning via post-transcriptional control of HSPA1A expression.

AIMS: Recent studies have identified the critical roles of nucleolin in a variety of cellular processes, including regulation of viral replication and tumour formation. However, the possible roles of nucleolin in myocardial preconditioning remain undefined. METHODS AND RESULTS: We used an in vivo rat myocardial ischaemic preconditioning (IP) model (four cycles of 5 min ischaemia and 10 min reperfusion) and cellular hydrogen peroxide preconditioning (H2O2-PC) models. We found that nucleolin mRNA and protein expression showed a time-dependent increase during the recovery of myocardial ischaemic preconditioning in rats and H2O2-PC in neonatal rat cardiomyocytes. Nucleolin overexpression enhanced the protective effects of H2O2-PC, whereas nucleolin ablation abrogated the H2O2-PC-mediated protection in cardiomyocytes. On the other hand, nucleolin overexpression increased the stabilization of the HSPA1A mRNA and the expression of HSPA1A protein in cardiomyocytes, whereas nucleolin ablation abrogated the up-regulation of HSPA1A induced by H2O2-PC in cardiomyocytes. An interaction between nucleolin and HSPA1A mRNA was further identified using the RNA-protein interaction studies. Reporter gene assays, which depended on the untranslated regions (UTR) of HSPA1A mRNA, revealed that the post-transcriptional regulation was mainly attributed to the 3' UTR. Finally, HSPA1A anti-sense oligonucleotides (asODNs) attenuated the protective effect of nucleolin in cardiomyocytes. CONCLUSION: These results indicate that nucleolin is up-regulated and involved in myocardial protection of ischaemic preconditioning via a post-transcriptional control of HSPA1A expression.

Nucleolin protects the heart from ischaemia-reperfusion injury by up-regulating heat shock protein 32.

AIMS: Nucleolin plays important roles in a variety of cellular processes. In this study, we aimed to investigate the role of nucleolin in cardiac ischaemia-reperfusion (I-R) injury. METHODS AND RESULTS: We investigated the expression pattern of nucleolin in hearts subjected to I-R, or neonatal rat cardiomyocytes subjected to hypoxia-re-oxygenation. We found that nucleolin expression was significantly down-regulated and the cleaved protein was present, both in vivo and in vitro. Gene transfection and RNA interference approaches were employed in cardiomyocytes to investigate the function of nucleolin. Over-expression of nucleolin was cytoprotective, whereas nucleolin ablation enhanced both hypoxia- and H2O2-induced cardiomyocyte death. Furthermore, transgenic mice with cardiac-specific over-expression of nucleolin were resistant to I-R injury as indicated by decreased cellular necrosis and decreased infarct size. The cardio-protective roles of nucleolin in cardiomyocytes, are attributable to the interaction of nucleolin with the mRNA of heat shock protein 32 (Hsp32), resulting in an increase of Hsp32 mRNA stability, and subsequent up-regulation of Hsp32 expression. The selective Hsp32 inhibitor, zinc protoporphyrin-IX, abrograted the cardiac protection mediated by nucleolin. CONCLUSION: This study has demonstrated that nucleolin is involved in the regulation of I-R-induced cardiac injury and dysfunction via the regulation of Hsp32, and may be a novel therapeutic target for ischaemic heart diseases.

MicroRNA profiling in denatured dermis of deep burn patients.

BACKGROUND: Denatured dermis is a part of the dermis in deep burn wound and has the ability to restore normal morphology and function. Skin grafting with the preservation of denatured dermis is a new kind of surgical procedure and has reported satisfactory clinical effects, such as lessened scar contracture and a better restoration of the appearance and function. However, the underlying mechanism of the recovery of denatured dermal function remains unclear. MicroRNAs (miRNAs) are a new class of regulatory noncoding single-stranded RNAs, which play a key role in normal development and physiology, as well as in disease development. This study analysed the profile of miRNAs in denatured dermis from patients and further investigated the possible roles of miRNAs played in the functional recovery of denatured dermis by prediction of the potential target genes of differentially expressed miRNAs. METHODS: The denatured dermis and paired normal skin were collected and analysed by miRNA array. The miRNA profiling results were validated by real-time reverse transcriptase polymerase chain reaction (RT-PCR), and bioinformatics' analysis was employed to further predict the miRNA targets. RESULTS: A total of 66 miRNAs were differentially expressed in denatured dermis compared with those in normal skin, among which 34 were down-regulated while 32 are up-regulated. The most significantly up-regulated miRNA was miR-663, and the most significantly down-regulated one was miR-203. Differentially expressed miRNAs were predicted to be related with several signalling pathways in wound healing. CONCLUSION: The differential miRNA expression identified in this study supplies experimental basis for further understanding the mechanisms of functional recovery of the denatured dermis.