Long non-coding RNA MALAT1 promotes Th2 differentiation by regulating microRNA-135b-5p/GATA-3 axis in children with allergic rhinitis.

Allergic rhinitis (AR) threatens patient survival. CD4+ T cells play key roles in AR progression. Long non-coding RNAs (lncRNAs) are key regulators of cell differentiation. Therefore, we investigated the molecular mechanism of the lncRNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in AR. Expression levels of MALAT1, microRNA (miR)-135b-5p, interleukin-4 (IL-4), and GATA-binding protein 3 (GATA-3) in the nasal mucosa of AR patients were quantified. CD4+ T cells were isolated from the peripheral blood of healthy volunteers and treated with ovalbumin (OVA) and Th2 inducers. After MALAT1 and miR-135b-5p levels changed in CD4+ T cells, the proportion of IL-4-expressing cells and the levels of IL-4 and GATA-3 in OVA-induced CD4+ T cells were determined. Binding relationships among MALAT1, miR-135b-5p, and GATA-3 were predicted and verified. Rescue experiments were performed to confirm the role of the MALAT1/miR-135b-5p/GATA-3 axis in Th2 differentiation of CD4+ T cells. MALAT1, IL-4, and GATA-3 expression was upregulated, whereas miR-135b-5p expression was downregulated, in patients with AR. MALAT1 knockdown or miR-135b-5p overexpression in CD4+ T cells notably decreased the proportion of IL-4-expressing cells and downregulated GATA-3 and IL-4 expression in OVA-induced CD4+ T cells. MALAT1 and GATA-3 exhibited competitive binding toward miR-135b-5p. MALAT1 facilitated CD4+ T cell Th2 differentiation via the miR-135b-5p/GATA-3 axis. MALAT1 facilitated AR development by facilitating CD4+ T cell Th2 differentiation via the miR-135b-5p/GATA-3 axis. This study may provide guidance for clinical treatment of AR.

[Predictive value of serum inhibin B levels as an indicator of the presence of testicular spermatozoa in nonobstructive azoospermia].

OBJECTIVE: To evaluate the predictive value of serum inhibin B (INH B) levels as an indicator of the presence of testicular spermatozoa in nonobstructive azoospermia. METHODS: Forty patients with nonobstructive azoospermia (NOA), 20 patients with obstructive azoospermia (OA), and 10 fertile volunteers were involved in this study. A chemoluminescence method was used to measure the levels of FSH; Inhibin B was analysed by using sandwich enzyme-linked immuno-sorbent assay. RESULTS: Patients with nonobstructive azoospermia has significantly higher levels of serum FSH [(21.34 +/- 12.15) IU/L] and significantly lower levels of inhibin B [(53.15 +/- 58.74) ng/L] than patients with obstructive azoospermia [FSH: (3.94 +/- 1.52) IU/L, INH B: (162.49 +/- 78.38) ng/L, P < 0.01] and fertile volunteers [FSH: (4.27 +/- 2.84) IU/L, INH B: (228.49 +/- 110.68) ng/L, P < 0.01]. Mean serum inhibin B were significantly higher in patients with nonobstructive azoospermia who had spermatozoa on TESE than in those in whom no spermatozoa was found on TESE [INHB: (90.31 +/- 72.18) ng/L vs (19.54 +/- 20.38) ng/L, r = 0.528, P < 0.01], but mean FSH levels did not have similar predictive power (P > 0.05). CONCLUSION: Serum INH B level seems to be more accurate than serum FSH in the prediction of presence of testicular spermatozoa in patients with nonobstructive azoospermia. Serum inhibin B determination may be substitute of TESE as a diagnostic index.

[Construction of SOD1 eukaryotic expression vector and its expression].

AIM: to construction of eukaryotic expression vector of the human SOD1 (Cu/Zn superoxide dismutase, SOD1) and expression in HeLa cells. METHODS: the open reading frame (ORF) of SOD1 was amplified from human peripheral blood by RT-PCR. TA cloning strategy was used to insert the target fragments into pUCm-T vector. The recombinant plasmid was identified and noted as pUCm-T-SOD1. Then, SOD1 was subcloned into pTracer-CMV/Bsd, a eukaryotic expression vector. The plasmid of pTracer-CMV/Bsd-SOD1 was sequenced and was introduced into HeLa cells by Lipofectamine(TM); 2000. The expression of green fluorescence protein (GFP) was observed by the fluorescence microscope. The expression of SOD1 was detected by RT-PCR and Western blot after screening by blasticidin for 4 weeks. RESULTS: the eukaryotic expression plasmid pTracer-CMV/Bsd-SOD1 was successfully constructed. The GFP was observed in transfected cells by the fluorescence microscope. The expression of SOD1 was detected in transfected cells by RT-PCR and Western blot. CONCLUSION: the recombinant eukaryotic expression vector of pTracer-CMV/Bsd-SOD1 has been constructed successfully which could express GFP and SOD1, respectively, providing a tool for further gene therapy study.

[Surface modification and DNA-binding assessment of nano-hydroxyapatite].

OBJECTIVE: To evaluate the impact of surface modification on the DNA-binding ability of nano-hydroxyapatite (nHA). METHODS: Chemical co-precipitation-hydrothermal synthesis was utilized to prepare the nHA particles, and polyethylenimine (PEI) was used for surface modification of the nHA. Transmission electron microscopic (TEM) observation and zeta potential detection of the nHA were carried out before and after surface modification. The abilities of the nanoparticles, at different pH values and different concentrations, for DNA-binding and DNA protection against nuclease digestion were assessed before and after surface modification by electrophoresis. RESULTS: TEM observation showed a short rod-like morphology of PEI-modified nHA with uniform particle size and good dispersion; the nHA without the modification tended to aggregate with poor dispersion. With a positive zeta potential, the PEI-modified nHA showed an obviously enhanced ability of DNA binding at different pH values and concentrations, with strong capacity to protect the DNA against Dnase I digestion. At the concentration of 250 µg/ml and a pH value of 7.0, the nHA-PEI showed an optimal efficiency of DNA-binding and DNA protection. CONCLUSION: nHA with surface modification by PEI can serve as an effective vector for DNA binding and transfer.