MiR-29b level-mediated regulation of Klotho methylation via DNMT3A targeting in chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) is a chronic lung disease characterized by chronic bronchitis and emphysema. Cigarette smoke extract (CSE) is the predominant cause of COPD. This study aimed to investigate the effects of miR-29b and their underlying mechanisms in a COPD cell model. MiR-29b and DNMT3A expression in lung tissue samples (taken at least 5 cm away from the tumor lesion) of NSCLC cases with smoking (n = 30), without smoking (n = 30), and with COPD (with smoking) (n = 30) was researched by qRT-PCR. A medium containing 10 % CSE was employed to induce murine alveolar macrophage MH-S cells to establish COPD cells. 5-Aza-cdr (5-AZA-2'-deoxycytidine) was used to block DNMT3A. The relationship and interaction between miR-29b and DNMT3A were validated through the dual luciferase reporter assay. The expression levels of macrophage M1 polarization marker proteins iNOS and TNF-α, DNMT3A, and Klotho protein were monitored using western blotting. The methylation levels of the miR-29b precursor gene and Klotho promoter were detected by quantitative methylation-specific PCR (MS-qPCR). The levels of IL-1ß, IL-6, and TNF-α in cell culture medium were detected via ELISA. It was found that the expression of miR-29b was downregulated, as a result of increased DNA methylation, and that of DNMT3A was upregulated in the lung tissues of NSCLC cases with COPD (with smoking). DNMT3A expression was negatively correlated with miR-29b expression in the lung tissues of NSCLC cases with COPD (with smoking). In addition, miR-29b expression was distinctly downregulated in CSE-induced MH-S cells and inhibited CSE-induced M1 polarization and inflammation. Importantly, DNMT3A was identified as a direct target gene of miR-29b. MiR-29b is negatively regulated by DNMT3A-mediated DNA methylation. Moreover, Klotho expression was downregulated and the Klotho promoter methylation level was increased in lung tissues of NSCLC cases with COPD (with smoking). The negative feedback between miR-29b and DNMT3A modulates CSE-induced M1 polarization and inflammation in macrophages as well as Klotho promoter methylation in CSE-mediated MH-S. Collectively, these findings indicate that the miR-29b level in COPD controls Klotho methylation via DNMT3, which maybe a promising target for the treatment of COPD.

Coexistence of anti-MDA5 and anti-PL-7 in a patient with dermatomyositis: A case report.

Coexisting anti-MDA5 and anti-PL-7 antibodies are extremely rare. Anti-MDA5 is associated with rapidly progressive interstitial lung disease (RP-ILD), while anti-PL-7 is often associated with chronic or subacute ILD and better outcomes than RP-ILD. We report a 41-year-old woman diagnosed with dermatomyositis (DM)-associated ILD positive for anti-MDA5 and anti-PL-7.

Purification and characterization of a novel angiotensin I-converting enzyme-inhibitory peptide derived from Alaska pollack skins.

Hydrolysates containing angiotensin I-converting enzyme (ACE)-inhibitory peptide were prepared from protein of Alaska pollack skins using alcalase and trypsin. The protein hydrolysate was separated by ultrafiltration, Sephadex G-25 gel filtration chromatography and reversed phase high-performance liquid chromatography (HPLC), from which a novel purified peptide was obtained. Both random coil structure and ß-sheet in the purified peptide were revealed in Fourier transform infrared spectrum. The amino sequence of the purified peptide was identified as GPLGVP, VLYPVK, VFLENVLR, and FEEF by HPLC-Q-TOF-MS (HPLC-quadrupole time-of-flight mass spectrometry). The peptide GPLGVP whose molecular weight was 538.31 Da showed the highest ACE inhibitory activity (IC50  = 105.8 µM). The purified peptide featured a noncompetitive inhibition kinetic mechanism was shown in the Lineweaver-Burk plots and was susceptible to enzymes as indicated in the studies on stability of gastrointestinal proteases. Moreover, the peptide GPLGVP can combine ACE catalytic pocket through hydrogen bonds and other forces with high binding power as disclosed in molecular docking simulation, which provides the inhibitory effect of GPLGVP on ACE.

[Signaling Pathways in the Pathogenesis of Acne Vulgaris].

Acne vulgaris is an inflammatory disease of hair follicle sebaceous gland units,with an incidence of up to 85% in adolescents.The pathogenesis is closely related to androgen,sebum secretion,lipophilic microbial infection,and immune-inflammatory reaction.This article reviews the signaling pathways related to acne from the aspects of inflammatory signaling pathways and sebum secretion pathways.

Endoplasmic reticulum stress links psoriasis vulgaris with keratinocyte inflammation.

INTRODUCTION: Endoplasmic reticulum stress (ERS) has been implicated in the pathogenesis of various inflammatory diseases. However, the role of ERS in psoriasis is still unclear. AIM: To examine ERS in psoriasis keratinocytes and to assess the association of ERS with skin inflammation response. MATERIAL AND METHODS: We investigated ERS in keratinocytes of normal skin, lesional and perilesional psoriasis vulgaris (PV) skin tissues using transmission electron microscope (TEM) examination, Western blot and immunostaining analysis. RESULTS: By TEM examination, we found that endoplasmic reticulum (ER) in psoriatic keratinocytes was ultrastructurally abnormal, with changes in ER morphology and the ER expansion. Using Western blot and immunostaining analysis, we showed that the expression of ERS-associated proteins, such as BiP, CHOP and XBP1, was enhanced in PV epidermis compared to the healthy skin. Moreover, abundant TNF-α protein was correlated to the increased BiP, CHOP and XBP1 expression in PV epidermis. CONCLUSIONS: Our findings demonstrate that PV keratinocytes have an increased ERS, which may contribute to the pathogenesis of PV.

Proteomic Analysis of Differentially Expressed Proteins in Intracranial Angiostrongylus cantonensis Larvae in Permissive and Non-Permissive Hosts.

Angiostrongylus cantonensis infection can lead to severe neuropathological damage caused by the development of these nematodes in the central nervous system after penetrating the blood-brain barrier. They commonly cause eosinophilic meningitis or meningoencephalitis in non-permissive hosts (e.g., mice). It has been shown that differences exist in the brains of permissive and non-permissive hosts during the larval development of A. cantonensis; however, the mechanism underlying the difference is not completely understood. This study analyzed and characterized the differentially expressed proteins in the intracranial A. cantonensis larvae in rat (ILR) and mouse (ILM) brains by using proteomics. We found that 29 proteins were differentially expressed: 12 of these proteins were highly expressed in ILR, whereas the remaining 17 proteins were highly expressed in ILM. Three protein spots were homologous to the actin-2, actin-1, and disorganized muscle protein 1 (dim-1) of Caenorhabditis elegans. In addition, proteomic analyses revealed that act-1 and act-2 were up-regulated in ILM compared to ILR, whereas dim-1 was down-regulated in ILM. Annotation using gene ontology revealed that act-1, act-2, and dim-1 were mainly associated with adenosine triphosphate (ATP) catabolic processes and ATP binding. Quantitative real-time polymerase chain reaction analyses of act-1 and dim-1 using the first internal transcribed spacers of A. cantonensis 18S ribosomal RNA (rRNA) was consistent with 2-dimensional gel electrophoresis (2-DE) and the sizes of these parasites; ILR was longer and wider than ILM. These results indicate that the differentially expressed proteins dim-1 and act-1 could be related to the development and pathogenicity of A. cantonensis in different hosts.

Syringotropic mycosis fungoides responding well to VELP chemotherapy: A case report.

Mycosis fungoides (MF), a low-malignant lymphoproliferative disorder, is the most common type of cutaneous T-cell lymphoma. The current study reported a case of syringotropic MF, a rare variant of MF, which presented with reactive B cell proliferation, lymphoid follicle formation, hair loss and lymphadenopathy. The clinical manifestations of the patient were MF-like lumps. Immunohistochemical staining of AE1/AE3 showed that there were abundant infiltrated lymphocytes surrounding the syringocystadenoma. In addition, the direction of the lymphocyte arrangement was consistent with the meandering direction of syringocystadenoma. The patient did not respond to 1-month narrowband (311-nm) ultraviolet therapy; however, a good response was obtained subsequent to one cycle of chemotherapy with vincristine sulfate, etoposide, L-asparaginase and prednisone acetate (know as the VELP regimen). After 7 days of VELP chemotherapy, the skin lesions were ameliorated, hair loss was improved and lymphadenopathy disappeared. No lymphadenopathy or new skin lesions were observed during 6 months of follow-up.

MiR-26b inhibits melanoma cell proliferation and enhances apoptosis by suppressing TRAF5-mediated MAPK activation.

Alterations in microRNA-26b (miR-26b) expression have been shown to participate in various malignant tumor developments. However, the possible function of miR-26b in human melanoma cells remains unclarified. In this study, quantitative polymerase chain reaction was used to explore the expression profiles of miR-26b in melanoma cells. The effect of miR-26b on cell viability was determined by using MTT assays and colony formation assay. The apoptosis levels were evaluated by using Annexin V/fluorescein isothiocyanate (FITC) apoptosis detection kit and the apoptosis cells were confirmed by Transmission Electron Microscopy (TEM). Luciferase reporter plasmids were constructed to confirm direct targeting. Our study found that the expression of miR-26b was downregulated in human melanoma specimens. Overexpression of miR-26b significantly increased the anti-proliferative effects and apoptosis in A375 and B16F10 melanoma cells. In addition, luciferase gene reporter assays confirmed that TRAF5 was a direct target gene of miR-26b and the anti-tumor effect of miR-26b in melanoma cells was significantly counteracted by treatment with TRAF5 overexpression. Furthermore, the molecular mechanisms underlying the tumor suppressor of miR-26b in malignant melanomas may be due to the dephosphorylation of MAPK pathway caused by the decrease in TRAF5 expression when miR-26b is up-regulated in melanoma cells. These findings indicate that miR-26b might influence TRAF5-MAPK signaling pathways to facilitate the malignant progression of melanoma cells.

Short-term clinical effects of photodynamic therapy with topical 5-aminolevulinic acid for facial acne conglobata: an open, prospective, parallel-arm trial.

BACKGROUND: Acne conglobata is hardly curable and easily leads to scar formation after treatment using traditional methods. AIM: To develop a novel way to treat acne conglobata. METHODS: Seventy-five patients with facial acne conglobata were included in this clinical study and divided into either a treatment group (n = 35) to receive photodynamic therapy (PDT) with topical 5% 5-aminolevulinic acid and red light once every 10 days for a month or a control group (n = 40) to receive a Chinese herbal medicine mask plus red light once per week for the same duration. Patients in both groups were given oral viaminate capsules, doxycycline, zinc gluconate, and topical metronidazole. Efficacy was evaluated with respect to symptom score, cure rate, and response rate up to 2 weeks following the final treatment, and time points for assessment included baseline (D0 ), the visit before each treatment (D10 and D20 for the treatment group, and D7 , D14 , and D21 for the control group), and 2 weeks after treatment (D34 for the treatment group and D35 for the control group). Safety was assessed by recording adverse effects. RESULTS: Treatment with PDT significantly improved acne lesions and reduced scar formation. The treatment group had a significantly lower symptom score, a higher cure rate, and response rate than the control group. No systemic side effects occurred. CONCLUSION: The treatment of acne conglobata with PDT is associated with a high cure rate, short treatment period, few side effects, and reduced scar formation. To the best of our knowledge, this is the first report on the treatment of acne conglobata with PDT.

[Expression of thrombospondin-1 in the lung of hypoxic pulmonary hypertension rats].

OBJECTIVE: To investigate the expression of thrombospondin-1(TSP-1) in the lung of hypoxia-induced pulmonary hypertension rats. METHODS: Thirty male Wistar rats were divided into two groups, pulmonary hypertension group and control group. The mice in experimental group were exposed to isobaric hypoxia for 3 weeks, and those in control group were exposed to air. The pulmonary artery pressure was measured by right cardiac catheterization. The expression of TSP-1 and TGF-beta1 in the lungs of rats were measured by immunohistochemical staining. The histological sections of the lungs were examined using a computerized image analyzer. RESULTS: After the induction of hypoxia for 3 weeks, the rats had pulmonary artery pressure increased with the thickening of the wall and the narrowing of the lumen of pulmonary arterioles. In the experimental group, the mean pulmonary artery pressure (mPAP) was (2.86 +/- 0.39) kPa, the index of right ventricular hypertrophy RV/(LV+S) was (43.53 +/- 3.38)%, the ratio of vascular wall thickness/vascular external diameter (WA%) was (55.09 +/- 12.38)%, and the ratio of vascular wall area/total vascular area (WT%) was (35.24 +/- 11.2)%, which all were significantly increased in comparison with those of control group [mPAP (1.35 +/- 0.28) kPa, RV/(LV+S) (23.68 +/- 3.48)%, WT% (23.63 +/- 9.74)%, WA% (41.62 +/- 12.83)%, respectively. P < 0.05). The positive staining of TSP-1 (1.32 +/- 0.04 vs. 0.96 +/- 0.03) and TGF-beta1 (1.38 +/- 0.05 vs. 1.04 +/- 0.04) in the wall of pulmonary arteriole of the rats exposed to hypoxia were significantly stronger than those of control rats (P < 0.01). CONCLUSION: The expression of TSP-1 appears to be increased in hypoxic pulmonary hypertension rats, which may contribute to the pathogenesis of hypoxic pulmonary vascular remodeling.

[The expression of thrombospondin-1 in serum and pulmonary arterioles of hypoxic pulmonary hypertension rats].

OBJECTIVE: To investigate the expression of thrombospondin-1 (TSP-1) in serum and pulmonary arterioles of rats with hypoxic pulmonary hypertension. METHODS: Twenty male Wistar rats were divided into two groups and exposed to air and isobaric hypoxia for 3 weeks respectively. The mean pulmonary artery pressure (mPAP) was measured by right cardiac catheterization. The rates of wall thickness/external diameter (WT%) and wall area/total vascular area (WA%) were calculated. The TSP-1 level in serum was measured by enzyme-linked immunosorbent assay. TSP-1 mRNA expression in lung tissue was evaluated by quantitative PCR. RESULTS: The pulmonary artery pressure increased in the hypoxia exposed rats. The chronic hypoxia also elicited the thicking of the wall and the narrowing of the lumen of pulmonary arterioles. It led to the increases of pulmonary artery pressure, the index of right ventricular hypertrophy [RV/(LV+S)], WA% and WT% compared to the controls [mPAP:(2.86 +/- 0.39) kPa vs. (1.35 +/- 40.28) kPa; RV/(LV+ S): (43.53 +/- 3.38)% vs. (23.68 +/- 3.48)%; WT%: (35.24 +/- 11.20)% vs. (23.63 +/- 9.74)%; WA%: (55.09 +/- 12.38)% vs. (41.62 +/- 12.83)% respectively, P<0.05]. In hypoxic group, the expression of TSP-1 mRNA in the lung was significantly up-regulated, the expression level of TSP-1 in serum was higher than that in control group (P<0.01). Linear correlation analysis showed that TSP-1 mRNA was positively associated with WT%, WA% and mPAP (r= 0.748, 0.686, 0.942 respectively, P<0.05). CONCLUSION: The TSP-1 may play an important role in the pathogenesis process of hypoxic pulmonary vascular remodeling and pulmonary hypertension.

Determination of molecular weight of hyaluronic acid by near-infrared spectroscopy.

This paper attempted the feasibility to determine the molecular weight of hyaluronic acid with near-infrared (NIR) diffuse reflectance spectroscopy. In this work, 46 experimental samples of hyaluronic acid powder were analyzed by partial least square (PLS) regression multivariate calibration method in the selected region of NIR spectra. The leave-one-out cross-validation method was used for the PLS model selection criterion. The accuracy of the final model was evaluated according to correlation coefficient of prediction set (Rp) and root mean square error of prediction set (RMSEP). The repeatability was verified through repeated measurement of spectra coupled with an appropriate chi-square test. Finally, the optimal calibration model was obtained with Rp=0.9814 and RMSEP=88.32 when using Savitzky-Golay first (SG-1st) derivative with 9 smoothing points spectral preprocessing method. The parameters above and repeatability of NIR spectroscopy obtained from chi-square test were both within the range of permissible error in factories. This study demonstrated that NIR spectroscopy was superior to conventional methods for the fast determination of molecular weight of hyaluronic acid.