Effect of An Atmospheric Plasma Jet on the Differentiation of Melanoblast Progenitor.

OBJECTIVE: Melanoblasts are the cell source of regeneration for pigment restoration. The ability to differentiate into mature melanocytes is the essential feature of melanoblasts in depigmentation diseases. Cold atmospheric plasma is an ionized gas with near-room temperature and highly reactive species that has been shown to induce stem cell differentiation. The aim of the study was to explore the effect of cold atmospheric plasma on the differentiation of melanoblast progenitor cells. METHODS: In this study, melanoblasts were exposed to the plasma jet and the cell morphology was observed. The cell cycle and cell proliferation were detected. Furthermore, the cell immunofluorescence and the detection of melanin particle and nitric oxide were carried out to investigate the differentiation of melanoblast progenitor cells. RESULTS: Cells that were treated with the plasma had longer and more synaptic structures, and the G1 phase of cell cycle was prolonged in the treated group. More melanin synthesis-related proteins and melanin particles were produced after plasma treatment. Nitric oxide was one of the active components generated by the plasma jet, and the nitric oxide content in the cell culture medium of the treated group increased. CONCLUSION: These results indicate that an increase in nitric oxide production caused by a plasma jet can promote cell differentiation. The application of plasma provides an innovative strategy for the treatment of depigmentation diseases.

Prognostic Value of ALP and LDH in Periampullary Carcinoma Patients Undergoing Surgery.

OBJECTIVE: Conversion of normal cells to cancer cells is often accompanied by abnormal synthesis of serum enzymes. Both alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) have been reported to have prognostic value in a variety of malignancies. The aim of this study was to investigate the effect of preoperative serum ALP and LDH levels on the prognosis of patients with periampullary carcinoma who underwent pancreatoduodenectomy (PD). METHODS: According to the preoperative ALP or LDH values, 856 cancer patients receiving PD treatment from January 2001 to January 2019 were divided into high-ALP group and low-ALP group or high-LDH group and low-LDH group. Statistical analysis was carried out to study the differences between the high-ALP and low-ALP groups or the high-LDH and low-LDH groups. Furthermore, the possibility of preoperative ALP or LDH as prognostic factor of periampullary carcinoma was investigated. RESULTS: In both the high-ALP and the high-LDH groups, the prognosis of patients with periampullary carcinoma who underwent PD was worse than that of the low-ALP and low- LDH group. Even through risk factor analysis, it was found that preoperative ALP and LDH could be independent prognostic factor for patients with periampullary carcinoma who underwent PD. CONCLUSION: Preoperative ALP or LDH is an independent risk factor for periampullary carcinoma.

CD36- and obesity-associated granulosa cells dysfunction.

Emerging evidence indicates that obesity impairs granulosa cell (GC) function, but the underlying mechanisms remain unclear. Gene expression profiles in GC of non-polycystic ovary syndrome (PCOS) obese (NPO), PCOS obese (PO), PCOS normal weight (PN) and non-PCOS normal weight (NPN) patients were analysed by microarray analysis. Compared with the NPN group, there were 16, 545 and 416 differently expressed genes in the NPO, PO and PN groups respectively. CD36 was the only intersecting gene, with greater than two fold changes in expression between the NPO versus NPN and PO versus NPN comparisons, and was not present in the PN versus NPN comparison. In addition, levels of CD36 protein were higher in GC from obese than normal weight patients. Furthermore, CD36 overexpression in a GC line inhibited cell proliferation, as determined by the cell counting kit-8 (CCK8) test, promoted cell apoptosis, as determined by flow cytometry, and inhibited the secretion of oestradiol by depositing triglyceride in cells and increasing cellular lipid peroxide levels. These adverse effects were reduced by sulfo-N-succinimidyloleate, a specific inhibitor of CD36. Together, the findings of this study suggest that obesity with and without PCOS should be regarded as separate entities, and that CD36 overexpression in GC of obese patients is one of the mechanisms by which obesity impairs GC function.

Alternative Oxidase Promotes Biofilm Formation of Candida albicans.

This study was designed to analyze the effect of the mitochondrial respiratory pathways of Candida albicans (C. albicans) on the biofilm formation. The 2, 3-bis (2-methoxy- 4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay was used to measure the metabolic activities of biofilms formed by the C. albicans which were cultured in the presence of respiratory pathways inhibitors. The biofilms formed by the wide type (WT), GOA7-deleted (GOA31), GOAV-reconstituted (GOA32), AOXla-deleted (AOX1) and AOXlb-deleted (AOX2) C. albicans strains were examined by the XTT reduction assay and fluorescence microscopy. The expression of adhesion-related genes BCR1, ALS1, ALS3, ECE1 and HWP1 in the biofilms formed by the above five C. albicans strains was detected by real time polymerase chain reaction. It was found that the metabolic activity of biofilms formed by C. albicans was decreased in the presence of alternative oxidase inhibitor whereas it was increased in the presence of classical mitochondrial respiratory pathway complex HI or complex IV inhibitor. AOX1 strain produced scarce biofilms interspersed with few hyphal filaments. Moreover, no significant changes in the expression of BCR1 and ALS3 were observed in the AOX1 strain, but the expression of ALSI and ECE1 was down-regulated, and that of HWP1 was up-regulated. These results indicate that both AOX1 and AOX2 can promote the biofilm formation. However, AOXla primarily plays a regulatory role in biofilm formation in the absence of inducers where the promoting effect is mainly achieved by promoting mycelial formation.

Activation of the HMGB1­TLR4­NF­κB pathway may occur in patients with atopic eczema.

High mobility group protein B1 (HMGB1) has been reported to serve important roles in various pathological conditions. Toll­like receptor 4 (TLR4), as one of the HMGB1 receptors, has been reported to be involved in the development of certain inflammatory diseases by activating nuclear factor NF­κ­B (NF­κB). However, there are few studies investigating the effects of HMGB1, TLR4 and NF­κB on human inflammatory dermatoses. In the present study, the distribution and characteristics of HMGB1, TLR4 and NF­κB p65 expression in psoriasis and atopic eczema (AE) were investigated. In addition, immunohistochemical analysis was performed to evaluate their expression and distribution in normal skin, and in patients with AE or psoria-sis. Spearman's correlation analysis was used to predicate their relevancy. The present study identified that the p65 level in epithelial nuclei in AE skin was increased compared with normal and psoriasis skin (P<0.01). The level of extracellular HMGB1 in AE skin was also increased compared with normal and psoriasis skin (P<0.01). Meanwhile, TLR4 expression on the epithelial membranes of AE skin was increased compared with psoriasis skin (P<0.01). Furthermore, the level of extracellular HMGB1 was positively correlated with epithelial membrane TLR4 (r=0.3856; P<0.05) and epithelial nuclear p65 (r=0.5894; P<0.01) in AE skin. These results indicated that the HMGB1­TLR4­NF­κB signaling pathway is activated in AE and may account for its pathogenesis, but not in psoriasis. Therefore, HMGB1, TLR4 and NF­κB p65 have the potential to be targets for the treatment of human inflammatory dermatoses, including AE.

[NF1 mutation analysis in a Chinese family with neuro- fibromatosis type].

A Chinese family affected with autosomal dominant disorder-neurofibromatosis type I was identified in this study. Linkage analysis was performed, and DNA sequencing for whole coding region of NF1 was carried out to identify the disease-causing mutation. The disease gene of the Chinese NF1 family was linked to NF1 locus, and a nonsense mutation, G1336X in the NF1 gene was identified. This mutation truncates the NF1 protein by 1 483 amino acid residues at the C-terminus, and is co-segregate with all the patients, but not present in unaffected individuals in the family. The present study demonstrated that G1336X mutation in the NF1 gene cause Neurofibromatosis type I in the family. To our knowledge, this mutation is firstly reported in Chinese population.

[Optimized expression of a single-chain Fv antibody against human asialoglycoprotein receptor and determination of its affinity constant].

AIM: To express and purify a single chain Fv antibody (scFv) C1 against human hepatic asialoglycoprotein receptor (ASGPR) and to determine affinity constant of the purified scFv C1. METHODS: The specific anti-ASGPR phage clone C1 was transfected into E. coli HB2151. The single colony was chosen to be inoculated into 2 x TY medium and shaken (250 r/min) overnight at 37 degrees C. After 1 in 100 dilution in 2 x TY medium and induced for secreted expression, scFv C1 was induced at different concentrations (0.25, 0.5 and 1.0 mmol/L IPTG) overnight at 37 degrees C, 25 degrees C and 20 degrees C, respectively. The supernatant was precipitated with saturated ammonium sulfate and its sediment was analyzed by SDS-PAGE. In addition, the sediment was resuspended in 30 mL PBS and dialyzed against PBS overnight at 4 degrees C. The expressed scFv C1 was purified by Ni(2+) chelating HiTrap HP column and the purity of the purified scFv C1 was identified by SDS-PAGE. Then affinity constant of scFv C1 was determined by noncompetitive enzyme immunoassay. RESULTS: After induced with 0.5 mmol/L IPTG overnight at 25 degrees C, the amount of expressed scFv C1 increased greatly and its relative molecular mass was about 28,000, and it existed in culture supernant in soluble form. The purity of scFv C1 by nickel-agarose column was above 95% and its yield was about 0.8 mg/L. The affinity constant of the purified scFv C1 was confirmed to be (2.31+/-0.36)x10(-7) mol/L. CONCLUSION: The E. coli HB2151 infected with phage C1 clones may express soluble scFv C1 with low affinity, which has potential applications to gene therapy of hepatocellular carcinoma.