Depression of LncRNA DANCR alleviates tubular injury in diabetic nephropathy by regulating KLF5 through sponge miR-214-5p.

OBJECTIVE: Diabetic nephropathy (DN) manifests a critical aspect in the form of renal tubular injury. The current research aimed to determine the function and mechanism of long non-coding ribonucleic acid (LncRNA) differentiation antagonising non-protein coding RNA (DANCR), with a focus on its impact on renal tubular injury. METHODS: Quantitative reverse transcription polymerase chain reaction was employed to analyze the RNA levels of DANCR in the serum of patients with DN or human proximal tubular epithelial cells (human kidney 2 [HK2]). The diagnostic significance of DANCR was assessed using a receiver operating characteristic curve. A DN model was established by inducing HK-2 cells with high glucose (HG). Cell proliferation, apoptosis, and the levels of inflammatory factors, reactive oxygen species (ROS), and malondialdehyde (MDA) were detected using the Cell Counting Kit - 8, flow cytometry, and enzyme-linked immunosorbent assay. The interaction between microRNA (miR)-214-5p and DANCR or Krüppel-like factor 5 (KLF5) was investigated using RNA immunoprecipitation and dual-luciferase reporter assays. RESULTS: Elevated levels of DANCR were observed in the serum of patients with DN and HG-inducted HK-2 cells (P < 0.05). DANCR levels effectively identified patients with DN from patients with type 2 diabetes mellitus. Silencing of DANCR protected against HG-induced tubular injury by restoring cell proliferation, inhibiting apoptosis, and reducing the secretion of inflammatory factors and oxidative stress production (P < 0.05). DANCR functions as a sponge for miR-214-5p, and the mitigation of DANCR silencing on HG-induced renal tubular injury was partially attenuated with reduced miR-214-5p (P < 0.05). Additionally, KLF5 was identified as the target of miR-214-5p. CONCLUSION: DANCR was identified as diagnostic potential for DN and the alleviation of renal tubular injury via the miR-214-5p/KLF5 axis, following DANCR silencing, introduces a novel perspective and approach to mitigating DN.

TaMPK2B, a member of the MAPK family in T. aestivum, enhances plant low-Pi stress tolerance through modulating physiological processes associated with phosphorus starvation defensiveness.

The mitogen-activated protein kinase (MAPK) cascades are present in plant species and modulate plant growth and stress responses. This study characterizes TaMPK2B, a MAPK family gene in T. aestivum that regulates plant adaptation to low-Pi stress. TaMPK2B harbors the conserved domains involving protein phosphorylation and protein-protein interaction. A yeast two-hybrid assay reveals an interaction between TaMPK2B and TaMPKK2 and between the latter and TaMPKKK;A, suggesting that all comprise a MAPK signaling cascade TaMPKKK;A-TaMPKK2-TaMPK2B. TaMPK2B expression levels were elevated in roots and leaves under a Pi starvation (PS) condition. Additionally, the induced TaMPK2B transcripts under PS in tissues were gradually restored following the Pi normal recovery condition. TaMPK2B overexpression conferred on plants improved PS adaptation; the tobacco lines with TaMPK2B overexpression enhanced the plant's dry mass production, Pi uptake capacity, root system architecture (RSA) establishment, and ROS homeostasis relative to wild type under PS treatment. Moreover, the transcripts of genes in phosphate transporter (PT), PIN-FORMED, and antioxidant enzyme (AE) families, including NtPT3 and NtPT4, NtPIN9, and NtMnSOD1 and NtPOD1;7, were elevated in Pi-deprived lines overexpressing TaMPK2B. Transgene analyses validated their functions in regulating Pi uptake, RSA establishment, and AE activities of plants treated by PS. These results suggest that TaMPK2B-mediated plant PS adaptation is correlated with the modified transcription of distinct PT, PIN, and AE genes. Our investigation suggests that TaMPK2B is one of the crucial regulators in plant low-Pi adaptation by improving Pi uptake, RSA formation, and ROS homeostasis via transcriptionally regulating genes associated with the above physiological processes.

Characterization on TaMPK14, an MAPK family gene of wheat, in modulating N-starvation response through regulating N uptake and ROS homeostasis.

KEY MESSAGE: Wheat MAPK gene TaMPK14 is N starvation response and is crucial in modulating plant low-N stress tolerance. Improving plant N use efficiency (NUE) contributes largely to the sustainable crop production worldwide. In this study, TaMPK14, a mitogen-activated protein kinase (MAPK) family gene in T. aestivum, was characterized for the role in mediating N starvation response. TaMPK14 harbors conserved domain/motifs specified by the plant MAPK proteins. In vitro assay for kinase activity of TaMPK14 validated its phosphorylation nature. TaMPK14 transcripts were upregulated in both roots and leaves under low-N treatment; moreover, the expression levels induced by N starvation were gradually restored following the N recovery progression. These results suggested transcriptional response of TaMPK14 upon the low-N stress. Compared with wild type (WT), the TaMPK14 overexpressing lines in N. tabacum displayed improved growth and N accumulation traits under deficient-N treatment, which indicated the crucial roles of the MAPK gene in mediating N starvation response. Additionally, the lines treated by N starvation were shown to be improved on cellular ROS homeostasis, displaying higher antioxidant enzymes (AE) activities and less ROS accumulative amount than WT. The transcripts of nitrate transporter gene NtNRT2.1 and those of AE genes NtSOD1, NtCAT1;2, and NtPOD4 were significantly upregulated in N-deprived TaMPK14 lines; overexpression of them conferred plants enhanced N uptake capacity and AE activities, respectively. Moreover, RNA-seq datasets generated from N-deprived transgenic lines contained numerous differential genes involving modulating various biological process, cellular component, and molecular function. Together, our investigation suggested that TaMPK14 improves plant N starvation response through transcriptional regulation of distinct NRT and AE genes as well as modulation of associated biological processes.

Use of DNA microarray chips for the rapid detection of Mycobacterium tuberculosis resistance to rifampicin and isoniazid.

The objective of the present study was to evaluate the potential development of DNA microarray chips to detect rifampicin (RFP) and isoniazid (INH) resistance in Mycobacterium tuberculosis (MTB), using samples from clinical tuberculosis (TB) patients in Soochow City, China. The sputum samples of 42 patients with TB in the Affiliated Hospital of Infectious Diseases of Soochow University (Soochow, China) were collected. The conventional Lowenstein-Jensen culture medium (Gold Standard) was used to assess drug sensitivity using the absolute concentration method. GeeDom MTB drug detection kits were also used to create a DNA microarray chip and examine the RFP-resistance associated gene mutation points rpoB-RRDR 511, 513, 516, 526, 531 and 533, and the INH-resistance associated gene mutation points katG315 and inhA-15 of the sputum samples. Compared with the results from the absolute concentration method, the susceptibility and specificity of RFP sensitivity detected by the DNA microarray chip were 92.8 and 93.8%, respectively. The susceptibility and specificity of INH sensitivity detected were 66.7 and 81%, respectively. The rpoB-RRDR 526, 531 mutations were the primary causes of MTB RFP resistance and the katG315 mutation was the primary cause of INH resistance. The detection of rpoB and katG gene mutation points by a DNA microarray chip may be used as a rapid, accurate and bulk clinical detection method for RFP and INH resistance in MTB. This is very valuable for the control of TB epidemics.

A prospective evaluation of the contrast, radiation dose and image quality of contrast-enhanced CT scans of paediatric abdomens using a low-concentration iodinated contrast agent and low tube voltage combined with 70% ASIR algorithm.

PURPOSE: To quantitatively and subjectively assess the image quality of and radiation dose for an abdominal enhanced computed tomography (CT) scan with a low tube voltage and a low concentration of iodinated contrast agent in children. METHODS: Forty-eight patients were randomised to one of the two following protocols: Group A (n=24, mean age 46.96±44.65 months, mean weight 15.71±9.11 kg, BMI 16.48±2.40 kg/m(2) ) and Group B (n=24, mean age 41.33±44.59 months, mean weight 18.15±17.67 kg, BMI 17.50±3.73 kg/m(2) ). Group A: 80 kVp tube voltage, 270 mg iodine (I)/mL contrast agent (Visipaque, GE Healthcare) and images were reconstructed using 70% adaptive statistical iterative reconstruction (ASIR). Group B: 100 kVp tube voltage, 370 mg I/mL contrast agent (Iopamiro, Bracco) and images were reconstructed using 50% ASIR. The volume of the contrast agent was 1.30 mL/kg in both Groups A and B. The degree of enhancement and noise in the abdominal aorta (AO) in the arterial phase (AP) and the portal vein (PV) in the portal venous phase (PVP) was measured; while the signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) for the AO and PV were calculated. A 5-point scale was used to subjectively evaluate the image quality and image noise by two radiologists with more than 10 years of experience. Dose-length product (DLP) (mGy-cm) and CTDIvol (mGy) were calculated. Objective measurements and subjective quality scores for the two groups were compared using paired t-tests and Mann-Whitney U tests, respectively. RESULTS: There was no significant difference in age, weight or body mass index (BMI) between the two groups (all P>.5). The iodine load in Group A (5517.3±3197.2 mg I) was 37% lower than that in Group B (8772.1±8474.6 mg I), although there was no significant difference between them (P=.111). The DLP and the CT dose index (CTDIvol ) for Group A were also lower than for Group B, but were not statistically significantly different (DLP, 104 mGy-cm±45.81 vs 224.5 mGy-cm±45.83; CTDIvol, 1.44 mGy±0.50 vs 2.08 mGy±1.87, all P>.05). The mean arterial and portal venous enhancement (255.33 HU±83.42, 146.41 HU±23.45, respectively), noise (AP 14.96 HU±2.09, PVP 16.30 HU±3.21), CNRs (AO 14.54±7.12, PV 5.07±1.73) and SNRs (AO 20.76±6.76, PV 12.43±3.24) for Group A were similar to Group B (enhancement: 226.55 HU±77.71, 138.69 HU±33.22; noise: 14.92 HU±3.12, 15.36 HU±3.48; CNRs: 12.96±7.14, 5.16±2.28; SNRs: 19.13±7.30, 12.69±4.22; all P>.05). The mean scores of the quality of the AP and PVP images in Group B were 4.31±0.53 and 4.35±0.52, respectively, while the scores obtained in Group A were 4.29±0.51 and 4.25±0.51; there were no statistically significant differences between the two groups. CONCLUSION: The scanning protocol using a low tube voltage (80 kVp) together with 70% ASIR and a low-concentration iodinated contrast agent (270 mg I/mL) enables a 37% reduction in iodine load and a 30% reduction in radiation dose while maintaining compatible image quality.

A new method of kidney biopsy using low dose CT-guidance with coaxial trocar and bard biopsy gun.

BACKGROUND: To explore a new method of kidney biopsy with coaxial trocar and bard biopsy gun under low dose computed tomography (CT)-guidance and evaluate its accuracy, safety, and efficacy. METHODS: Sixty patients underwent renal biopsy under CT-guidance. They were randomly divided into two groups: group I, low dose CT-guided (120 kV and 25 or 50 mAs) and group II, standard dose CT-guided (120 kV and 250 mAs). For group I, the coaxial trocar was accurately placed adjacent to the renal capsule of the lower pole, the needle core was removed, and samples were obtained with a bard biopsy gun. For group II, the coaxial trocar was not used. Total number of passes, mean biopsy diameter, mean glomeruli per specimen, mean operation time, mean scanning time, and mean radiation dose were noted. Dose-length product (DLP) was used to calculate the radiation doses. After 24 hours of the biopsy, ultrasound was repeated to identify any subcapsular hematoma. RESULTS: Success rate of biopsy in group I was 100% while using low dose CT-guidance along with coaxial trocar renal. There was no statistic differences bewteen group I and II in the total number of passes, mean biopsy diameter, mean glomeruli per specimen and mean time of operation and CT scanning. The average DLP of group I was lower as compared to the value of group II (p <0.05). CONCLUSIONS: Kidney biopsy using coaxial trocar and bard biopsy gun under low dose CT was an accurate, simple and safe method for diagnosis and treatment of kidney diseases. It can be used for repeat and multiple biopsies, particularly suitable for obese and renal atrophy patients in whom the kidneys are difficult to image.