Diabetic polyneuropathy and carpal tunnel syndrome together affect hand strength, tactile sensation and dexterity in diabetes patients.

AIMS/INTRODUCTION: Carpal tunnel syndrome (CTS) and diabetic polyneuropathy (DPN) can occur together, and this concomitance is thought to be higher in diabetes patients. We aimed to examine and compare hand function in type 2 diabetes mellitus patients without CTS and DPN (CTS-DPN-), patients with CTS without DPN (CTS+DPN-), patients with DPN without CTS (CTS-DPN+), and patients with CTS and DPN (CTS+DPN+). MATERIALS AND METHODS: A total of 161 type 2 diabetes mellitus patients underwent physical examination and electrodiagnostic tests. Grip and pinch strengths, tactile sensory thresholds were measured for each participant. Purdue pegboard test was used in evaluating the hand dexterity of the participants. RESULTS: Of the 161 type 2 diabetes mellitus participants, 36 (22.4%) had both CTS and DPN. CTS participants had lower grip (26.6 ± 10.6 vs 35.2 ± 14.3, P < 0.001) and pinch (6.3 ± 2.6 vs 7.5 ± 2.9, P = 0.026) strengths compared with non-CTS participants, whereas DPN participants had elevated tactile sensory thresholds of both the second (2.8 [2.8-3.6] vs 2.4 [2.4-2.8], P < 0.001) and the fifth (2.8 [2.8-3.6] vs 2.4 [2.4-2.8], P < 0.001) fingers compared with non-DPN participants. The CTS+DPN+ group had lower Purdue pegboard test scores than other groups. Grip (r = 0.482, 0.530, 0.467, 0.498, all P < 0.001) and pinch (r = 0.246, P = 0.003; r = 0.265, P = 0.001; r = 0.264, P = 0.001; r = 0.235, P = 0.005) strengths were positively correlated with Purdue pegboard test scores, whereas tactile sensory thresholds were negatively correlated with Purdue pegboard test scores (r = -0.447 to -0.359, all P < 0.001). CONCLUSION: Type 2 diabetes mellitus patients with both DPN and CTS had lower grip and pinch strengths and decreased tactile sensation, both of which were correlated with poorer hand dexterity.

LncRNA TRHDE-AS1 inhibit the scar fibroblasts proliferation via miR-181a-5p/PTEN axis.

Hypertrophic scar (HS), a fibroproliferative disorder caused by abnormal wound healing after skin injury, which is characterized by excessive deposition of extracellular matrix and invasive growth of fibroblasts. Recent studies have shown that some non-coding RNA implicated the formation of HS, but the mechanism remains unclear. In this study, we found that lncRNA TRHDE-AS1 was downregulated in HS tissues and HSFs, and the level of lncRNA TRHDE-AS1 negatively correlated with the level of miR-181a-5p in HS tissue and HSFs. Overexpressed lncRNA TRHDE-AS1 significantly suppressed miR-181a-5p level, while promoted HSFs apoptosis and inhibited HSFs proliferation. Further study shown that PTEN was a direct target of miR-181a-5p, and lncRNA TRHDE-AS1 served as a molecular sponge for miR-181a-5p to regulate the expression of PTEN. Overexpression of PTEN could eliminate lncRNA TRHDE-AS1-mediated proliferation suppression of HSFs. In conclusion, our study suggested that lncRNA TRHDE-AS1/miR-181a-5p/PTEN axis plays an important role in promoting hypertrophic scar formation, which may be effectively used as a therapeutic target for hypertrophic scar treatment.

LncRNA SNHG15 Knockdown Protects Against OGD/R-Induced Neuron Injury by Downregulating TP53INP1 Expression via Binding to miR-455-3p.

Cerebral ischemia-reperfusion (I/R) injury is the common symptom of ischemic stroke, which poses a heavy burden to human health. Long non-coding RNA (lncRNA) is indicated to be a critical regulator in cerebral ischemia. This study aims to reveal the effects of lncRNA small nucleolar RNA host gene 15 (SNHG15) on oxygen-glucose deprivation and reoxygenation (OGD/R)-induced neuron injury and underlying mechanism. The expression levels of SNHG15, microRNA-455-3p (miR-455-3p) and tumour protein p53 inducible nuclear protein 1 (TP53INP1) mRNA were determined by quantitative real time polymerase chain reaction in P12 cells. The protein levels of TP53INP1, cleaved caspase-3, caspase-3, B-cell lymphoma-2 and BCL2-associated x protein (Bax) were detected by western blot in P12 cells. Cell viability and apoptosis were revealed by cell counting kit-8 assay and flow cytometry analysis, respectively, in P12 cells. Caspase-3 activity, the levels of tumor necrosis factor-α and interleukin-1ß and the production of reactive oxygen species (ROS) were severally determined by caspase-3 activity assay, Enzyme-linked immunosorbent assay and ROS detection assay in P12 cells. The binding relationship between miR-455-3p and SNHG15 or TP53INP1 was predicted by starbase online database, and identified by dual-luciferase reporter, RNA pull-down or RNA immunoprecipitation assay. SNHG15 expression and the mRNA and protein levels of TP53INP1 were dramatically upregulated, while miR-455-3p expression was apparently downregulated in OGD/R-induced PC12 cells. SNHG15 silencing hindered the effects of OGD/R treatment on cell viability, apoptosis, inflammation and oxidative in PC12 cells; however, these impacts were restored after miR-455-3p inhibitor transfection. Additionally, SNHG15 acted as a sponge of miR-455-3p and miR-455-3p bound to TP53INP1. SNHG15 contributed to OGD/R-induced neuron injury by regulating miR-455-3p/TP53INP1 axis, which provided a novel insight to study lncRNA-directed therapy in ischemia stoke.

MicroRNA-138 Regulates T-Cell Function by Targeting PD-1 in Patients with Hepatitis B Virus-Related Liver Diseases.

OBJECTIVE: T-cell exhaustion in hepatitis B virus (HBV) infection, which results from upregulation of programmed cell death-1 (PD-1), leads to persistent HBV infection and related disease progression. Therefore, agents targeting PD-1 may prove beneficial in the treatment of this condition. MicroRNA-138 (miR-138) possesses an anti-tumor ability in that it targets immune checkpoints, including PD-1. However, the function and underlying mechanisms of miR-138 in patients with HBV infection remains unclear. METHODS: Specimens were collected from healthy volunteers (n = 43) and patients with chronic hepatitis B (CHB; n = 52), liver cirrhosis (LC; n = 26), and hepatocellular carcinoma (HCC; n = 31); carriers of HBV who were asymptomatic (n = 51); and patients with CHB receiving antivirus treatment (n = 11). These specimens were then used to study the expression and relationship among miR-138, PD-1, and HBV DNA viral load. To investigate the role of miR-138 in regulating PD-1 expression and determine the effect of miR-138 in regulating T-cell function, a luciferase assay and a transfection assay were each performed with primary CD3+ T cells. RESULTS: We found that PD-1 was upregulated and miR-138 was downregulated in patients with CHB, LC, and HCC. Correlations analysis revealed that PD-1 expression was positively correlated with HBV DNA viral load whereas miR-138 was negatively correlated. Luciferase assay results showed that miR-138 directly inhibited PD-1 expression by interacting with the 3'-untranslated region of PD-1. As a result of miR-138 overexpression in primary T cells, PD-1 in these T cells was downregulated and antivirus cytokines secreted by T cells were significantly upregulated. In addition, the expression levels of PD-1 and miR-138 were reversed in patients with CHB who received antivirus treatments. CONCLUSION: Results showed that miR-138 can promote T-cell responses within patients with HBV infection by inducing a PD-1 blockade. Such an effect suggests that miR-138 may serve as a new therapeutic target for the treatment of HBV infection.

Ablation of TMEM126B protects against oxygen-glucose deprivation/reoxygenation-induced injuries of PC12 cells via maintaining mitochondrial anti-apoptotic functions.

Ischemia reperfusion (I/R) injury is a key contributing factor to the pathogenic mechanism involved in cerebral infarction. Transmembrane protein 126b (TMEM126B), a mitochondrial complex I assembly factor, has been reported to have an intimate association with disease progression, but is little known in ischemia stroke. The present study was designed to explore the effects of TEME126B on oxygen-glucose deprivation/reoxygenation (OGD/R)-induced neuronal PC12 cells. The mRNA level of TMEM126B was determined using qRT-PCR. The levels of ROS, MDA, and SOD, as well as inflammatory cytokines, were measured using corresponding commercial kits. Cell apoptosis rate was assayed by flow cytometry analysis, and the apoptosis-related proteins were measured using western blotting. ATP production measured by colorimetric reaction and mitochondrial membrane potential measured by JC-1 staining were conducted to determine mitochondrial dysfunction. The results showed that TMEM126B was upregulated upon I/R injury in vitro and in clinical, and was positively corrected with the degree of oxidative stress. TMEM126B knockdown significantly reduced oxidative stress and inflammation in OGD/R-induced PC12 cells. TMEM126B knockdown also attenuated cell apoptosis rate, accompanied with increased expressions of Bcl-2, XIAP and cleaved PARP-1, and decreased expressions of Bax, cleaved caspase 3 and cleaved caspase 9. Furthermore, TMEM126B knockdown exhibited cytoprotective roles through alleviating mitochondrial dysfunction, as assessed by ATP production and mitochondrial membrane potential. Collectively, this study indicates that TMEM126B knockdown protects against OGD/R-induced neuronal injuries through relieving oxidative stress, inflammation, apoptosis and mitochondria dysfunction, which provides a promising target for ischemic stroke treatment.

Clinical, neuroelectrophysiological and muscular pathological analysis of chronic progressive external ophthalmoplegia.

The aim of the present study was to explore the clinical, neuroelectrophysiological and muscular pathological characteristics of chronic progressive external ophthalmoplegia (CPEO) and to improve the understanding of CPEO. Clinical manifestations, neuroelectrophysiology and pathological features of muscle biopsies from 12 patients with CPEO were retrospectively analyzed. The average age of onset for the 12 patients (6 males and 6 females) was 17.2 years. All patients had different degrees of blepharoptosis. A total of 11 patients experienced ocular dyskinesia, but diplopia was rare. Electrophysiological testing in 12 patients revealed abnormal changes in 6 patients, including 4 patients with a myogenic lesion, 1 patient with a neurogenic lesion, and 1 patient with mixed myogenic/neurogenic lesions. Two patients had slow sensory nerve conduction velocity. Muscle biopsies in 12 patients demonstrated ragged-red, irregular and broken fibers in 11 patients through Gomori trichrome and hematoxylin and eosin (H&E) staining, increased lipid levels in some muscle fibers in 4 patients through Οil Ρed O staining and abnormal distribution of type I and II muscle fibers in 3 patients through ATPase staining. Electron microscopy in 5 patients showed an increased number of mitochondria and abnormal mitochondrial aggregation between submucosa and myofibrils in 4 patients. These findings suggest that the possibility of CPEO should be considered if patients present with obvious extraocular muscle paralysis without diplopia. Furthermore, the identification of ragged-red fibers by Gomori trichrome and H&E staining of muscle biopsies from patients is an important basis for the diagnosis of CPEO.

Stat3 promotes invasion of esophageal squamous cell carcinoma through up-regulation of MMP2.

Stat3 alters the expression of its downstream genes and is associated with tumor invasion and metastasis in several human cancers. Its role in esophageal squamous cell carcinoma (ESCC) has not been well characterized. We examined the tumor sections of 100 cases of ESCC by immunohistochemistry and observed significant overexpression of Stat3 in the cytoplasm of 89% of ESCC cells and of phosphorylated Stat3 (p-Stat3) in the nuclei of 71% of ESCC when compare with normal esophageal mucosa (72%, p = 0.02; and 31%, p = 0.001). Overexpression of Stat3 and p-Stat3 positively correlated with that of matrix metalloproteinase-2 (MMP2), a known regulator for cell migration, in 65% of ESCC while only 26% shown in benign esophageal mucosa. To further investigate the association of Stat3 with tumor metastasis in vitro, invasion of EC-1 cells (a human ESCC cell line) were investigated with Boyden chambers. The results showed that transfection of Stat3 not only promoted invasion of EC-1 cells but also significantly induced MMP2 expression in a dose-dependent manner. In contrast, suppressing expression of endogenous Stat3 mRNA and protein by Stat3 siRNA significantly reduced EC-1 cell invasion and MMP2 expression. A high-affinity Stat3-binding element was localized to the positions of 648-641 bp (TTCTCGAA) in the MMP2 promoter with electrophoretic mobility shift assay. Our results suggest that Stat3, p-Stat3, and MMP2 were overexpressed in ESCC and associated with invasion of ESCC; and Stat3 up-regulated expression of MMP2 in ESCC through directly binding to the MMP2 promoter.

S100A4 mediated cell invasion and metastasis of esophageal squamous cell carcinoma via the regulation of MMP-2 and E-cadherin activity.

It is well documented that S100A4 is upregulated in a large amount of invasive tumors and plays a pivotal role in tumor invasion and metastasis. However, the precise role and mechanism S100A4 exerts in the invasion and metastasis of esophageal squamous cell carcinoma (ESCC) have not been fully elucidated to date. Our data demonstrated that S100A4 was overexpressed in human ESCC tissues, especially in ESCC with poor differentiation, deep invasion and lymph node metastasis. Subsequently, the knockdown of S100A4 by RNAi in ESCC cell line (EC-1) could reduce cell invasion, metastasis and proliferation ability in vitro. Most importantly, S100A4 regulated MMP-2 positively and E-cadherin negatively in vivo and in vitro to some extent. Our results suggest that S100A4 is an important factor in the invasion, metastasis and proliferation of ESCC and may control invasion and metastasis at least in part through the regulation of MMP-2 and E-cadherin activity. S100A4 may serve as a biomarker for progression of ESCC and a potential molecular target for biotherapy of ESCC.

[Expression of S100A4 in esophageal squamous cell carcinoma and its relation to tumor invasion and metastasis].

OBJECTIVE: To study the role of S100A4 in the carcinogenesis, development, invasion and metastasis of esophageal squamous cell carcinoma. METHODS: Immunohistochemistry was used to detect the expressions of S100A4, MMP-2 and E-cadherin proteins in 100 cases of surgically resected esophageal squamous cell carcinoma specimens. RT-PCR and Western blot were used to detect the expressions of S100A4 mRNA and protein in esophageal squamous cell carcinoma line EC-1 and TE-1. Boyden-chamber model in vitro was utilized to detect the invasion ability of EC-1 and TE-1 cells. RESULTS: The positivity rate of S100A4 protein was 52.0% was in esophageal carcinoma tissues, significantly higher than that in normal tissues (26.0%) (P<0.01). The expression of S100A4 was related to tumor grading, invasive depth and lymph node metastasis (P<0.05). In esophageal carcinoma, the expression of S100A4 was positively correlated to MMP-2 expression (P<0.01), but inversely to E-cadherin expression (P<0.05). The expressions of S100A4 mRNA (0.894-/+0.021) and protein (0.897-/+0.053) in EC-1 cells were significantly higher than those in TE-1 (0.812-/+0.040 and 0.645-/+0.089, respectively, P<0.01), and the invasion ability of EC-1 cells was significantly higher than that of TE-1 cells (91.00-/+17.44 vs 61.80-/+11.10, P<0.01). CONCLUSION: The overexpression of S100A4 in esophageal squamous cell carcinoma tissue and highly invasive EC-1 cells may contribute to the carcinogenesis, development, invasion and metastasis of esophageal squamous cell carcinoma.

[Inhibitory effect and molecular mechanism of silencing S100A4 gene on the growth of transplanted tumor of human esophageal carcinoma EC-1 cells in nude mice].

OBJECTIVE: To study the effect of S100A4siRNA on tumor growth of xenografted human esophageal squamous cell carcinoma (ESCC) cell line EC1 in nude mice and explore its related molecular mechanism. METHODS: The xenografted tumor model was established in nude mice, and S100A4siRNA chemically synthesized was used to transfect the xenografted nude mice. The tumor growth was observed. The mRNA and protein expressions of S100A4, MMP-2 and E-cadherin in tumor after transfection with S100A4siRNA were detected by RT-PCR and immunohistochemistry. RESULTS: The tumor volume of S100A4siRNA transfection group was lower than that of nonsense siRNA transfection group and blank control group (P < 0.05), and tumor inhibitive ratio of S100A4siRNA group was higher than that of nonsense siRNA transfection group (P < 0.05). Furthermore, the mRNA and protein expressions of S100A4, MMP-2 in S100A4siRNA group were lower than those in nonsense siRNA group and control group (P < 0.05), the mRNA and protein expressions of E-cadherin in S100A4siRNA group were higher than those in nonsense siRNA group and control group (P < 0.05). CONCLUSION: S100A4siRNA could effectively lead to growth inhibition of xenografted human esophageal tumor in nude mice, with down regulation of MMP-2 and up regulation of E-cadherin, which provides theoretical basis for molecular therapy of ESCC.

[Effect of KISS-1 on invasive potential and proliferation of esophageal squamous carcinoma cell line EC-1].

OBJECTIVE: To investigate the effect of KISS-1 expression on the potential of invasion and proliferation of esophageal squamous carcinoma cell EC-1. METHODS: Protein and mRNA expressions of KISS-1 were evaluated by Western blot and RT-PCR in four esophageal carcinoma cell lines (EC-1, Eca109, EC9706 and TE-1). Using liposome-mediated transfection, an eukaryotic expression vector (pcDNA3.1-KISS-1) of KISS-1 gene was transfected into EC-1 cells. Boyden chamber model, MTT and clone formation assay were used to detect the potential of invasion and proliferation. RESULTS: Western blot and RT-PCR showed a baseline low level of expression of KISS-1 protein (0.715 +/- 0.109) and mRNA (0.670 +/- 0.176) in EC-1 cells. pcDNA3.1-KISS-1 expression vector was successfully transfected into EC-1 cells. Western blot and RT-PCR showed that the expression of KISS-1 protein (1.143 +/- 0.218) and mRNA (0.877 +/- 0.162) in EC-1 cells transfected with pcDNA3.1-KISS-1 were significantly higher than those transfected with the control vector pcDNA3.1 (0.745 +/- 0.130, 0.685 +/- 0.128; t = 3.850, 2.481, P < 0.05) and the control cells (0.855 +/- 0.184, 0.677 +/- 0.138; t = 2.275, 2.306, P < 0.05). Boyden chamber analysis showed that the invasiveness of the cells transfected with KISS-1 at 24 h (91.8 +/- 11.7), 48 h (117.8 +/- 11.1) and 72 h (139.2 +/- 11.8) were significantly reduced than that of the cells transfected with the control vector pcDNA3.1 (118.1 +/- 14.7, 141.7 +/- 13.2, 162.2 +/- 22.7; t = 3.153, 4.215, 3.569, P < 0.01) and the control cells (112.2 +/- 15.6, 138.1 +/- 13.0, 162.3 +/- 14.0; t = 4.154, 3.797, 2.702, P < 0.05). MTT showed that the proliferation potential of cells after transfection with KISS-1 at 48 h (0.517 +/- 0.127) and 72 h (0.394 +/- 0.137) were significantly reduced than that of cells transfected with the control vector pcDNA3.1 (0.636 +/- 0.186, 0.513 +/- 0.150; t = 2.054, 2.709, P < 0.05) and the control cells (0.646 +/- 0.135, 0.511 +/- 0.153; t = 2.276, 2.205, P < 0.05). Clone formation assay suggested that cells transfected with KISS-1 (157.2 +/- 36.4) showed significantly decreased clone formation than cells transfected with the control vector pcDNA3.1 (236.3 +/- 78.1; t = 3.441, P < 0.01) and the control cells (242.5 +/- 48.6; t = 2.250, P < 0.05). CONCLUSION: KISS-1 gene inhibits the potential of invasion and proliferation of EC-1 cells.