•The conventional methods and Next-generation sequencing to diagnose thalassemia have limitations.•Long-read SMRT sequencing has been demonstrated to be more effective and accurate than conventional ...methods, especially for rare and complicated thalassemia variants.•A 762 bp deletion and a 342 bp insertion in α-globin gene cluster were identified by SMRT sequencing and reported for the first time.•Subjects with other rare mutations including α Fusion mutation, α-triplicates, α-quadruplicates and conversion of HBA2 to HBA1 was precisely identified by SMRT sequencing.
Gap- polymerase chain reaction (PCR), reverse dot-blot assay (RDB), real-time PCR based multicolor melting curve analysis (MMCA assay), multiplex ligation-dependent probe amplification (MLPA) and Sanger sequencing are conventional methods to diagnose thalassemia but all of them have limitations. In this study, we applied single-molecule real-time (SMRT) sequencing following multiplex long-range PCR to uncover rare mutations in nine patients and their family members. The patients with different results between Gap-PCR and MMCA assay or with phenotype not matching genotype were included. Using SMRT sequencing, we first identified the carriers with αααanti3.7/HKαα, -α762bpα/αα (chr16:172,648–173,409), ααfusion/αQSα (in a trans configuration), two cases with novel gene rearrangements and another case with a novel 341 bp insertion in α-globin gene cluster, respectively. One carrier with --SEA/αααanti4.2, and two carriers with the coexistence of globin variant and an α-globin gene duplication were also found. Most importantly, we could determine two defects in α-globin gene cluster being a cis or trans configuration in a single test. Our results showed that SMRT has great advantages in detection of α-globin gene triplications, rare deletions and determination of a cis or trans configuration. SMRT is a comprehensive and one-step method for thalassemia screening and diagnosis, especially for detection of rare thalassemia mutations.
Background Abelmoschus manihot , a single medicament of traditional Chinese medicine, has been widely used to treat kidney disease. This is the first randomized controlled clinical trial to assess ...its efficacy and safety in patients with primary glomerular disease. Study Design Prospective, open-label, multicenter, randomized, controlled, clinical trial. Setting & Participants From May 2010 to October 2011, a total of 417 patients with biopsy-proven primary glomerular disease from 26 hospitals participated in the study. Interventions A manihot in the form of a huangkui capsule, 2.5 g, 3 times per day; losartan potassium, 50 mg/d; or combined treatment, a huangkui capsule at 2.5 g 3 times per day, was combined with losartan potassium, 50 mg/d. The duration of intervention was 24 weeks. Outcomes & Measurements The primary outcome was change in 24-hour proteinuria from baseline after treatment. Change in estimated glomerular filtration rate (eGFR) from baseline after treatment was a secondary outcome. The 24-hour proteinuria was measured every 4 weeks and eGFR was measured at 0, 4, 12, and 24 weeks. Results Mean baseline urine protein excretion was 1,045, 1,084, and 1,073 mg/d in the A manihot , losartan, and combined groups, respectively, and mean eGFR was 108, 106, and 106 mL/min/1.73 m2 , respectively. After 24 weeks of treatment, mean changes in proteinuria were protein excretion of −508, −376, and −545 mg/d, respectively ( P = 0.003 for A manihot vs losartan and P < 0.001 for the combined treatment vs losartan). Mean eGFR did not change significantly. The incidence of adverse reactions was not different among the 3 groups ( P > 0.05), and there were no severe adverse events in any group. Limitations Results cannot be generalized to those with nephrotic syndrome or reduced eGFR. Conclusions A manihot is a promising therapy for patients with primary kidney disease (chronic kidney disease stages 1-2) with moderate proteinuria.
Rice false smut has become an increasingly serious disease in rice （Oryza sativa L.） production worldwide. The typical feature of this disease is that the fungal pathogen Ustilaginoidea virens （Uv） ...specifical y infects rice flower and forms false smut bal , the ustiloxin-containing bal-like fungal colony, of which the size is usual y several times larger than that of a mature rice seed. However, the underlying mechanisms of Uv-rice interac-tion are poorly understood. Here, we applied time-course microscopic and transcriptional approaches to investigate rice responses to Uv infection. The results demonstrated that the flower-opening process and expression of associated transcription factors, including ARF6 and ARF8, were inhibited in Uv-infected spikelets. The ovaries in infected spikelets were interrupted in fertilization and thus were unable to set seeds. However, a number of grain-fil ing-related genes, including seed storage protein genes, starch anabolism genes and endosperm-specific transcription factors （RISBZ1 and RPBF）, were highly transcribed as if the ovaries were fertilized. In addition, critical defense-related genes like NPR1 and PR1 were downregulated by;Uv infection. Our data imply that Uv may hijack host nutrient reservoir by activation of the grain-fil ing network because of growth and formation of false smut bal s.
Type 2 diabetes mellitus (T2DM) is a major contributor to peripheral artery disease (PAD), especially in cases that advance to critical limb ischemia (CLI). Accumulating evidence indicates that ...miRNAs play an important role in the development of PAD and T2DM. Due to the limited value of current diagnostic methods for CLI in T2DM patients, we compared the miRNA expression profiles of Chinese T2DM patients with or without CLI to find out whether distinctive miRNAs could serve as potential diagnostic biomarkers. We statistically identified 7 miRNAs (hsa-miR-200b-3p, hsa-miR-2115-3p, hsa-miR-431-5p, hsa-miR-486-5p, hsa-miR-210-3p, hsa-miR-1264, hsa-miR-323b-5p) which were up-regulated in the CLI group, whereas other 4 miRNAs (hsa-miR-5579-3p, hsa-miR-665, hsa-miR-4285, hsa-miR-500a-3p) were down-regulated. Our validation test suggested a relatively high diagnostic accuracy of serum hsa-miR-323b-5p levels for the detection of CLI in T2DM patients, with a sensitivity of 62.67% and a specificity of 80.65%. The area under the curve (AUC) for miR-323b-5p + confounding risk factors was 0.94 (95% CI: 0.884-0.994, P < 0.001), which was higher than that for miR-323b-5p. Taken together, our results indicate that circulating hsa-miR-323b-5p could be a promising serum biomarker for the diagnosis of critical limb ischemia in type 2 diabetic patients.
Friction stir welding was performed on AA2219-T8 plates with 6.31 wt pct Cu. The thermal cycles were measured in different regions of the joint during welding. Differential scanning calorimetry and ...transmission electron microscopy were utilized to analyze the evolution of precipitates in the joint. The relationships between welding peak temperature, precipitate evolution, and microhardness distribution are discussed. The temperature in the heat-affected zone (HAZ) ranged from 453 K to 653 K (180 °C to 380 °C). The
θ″
and some
θ′
phases redissolved into the HAZ matrix, while the rest of the
θ′
phases coarsened. In the thermomechanically affected zone (TMAZ), the temperature range was from 653 K to 673 K (380 °C to 400 °C), causing both
θ″
phase and
θ′
phase to redissolve. In the weld nugget zone (WNZ), all the
θ″
,
θ′
, and some of the
θ
phase (Al
2
Cu) redissolved. Abnormal
θ
particles were observed in the WNZ, including agglomerated
θ
with sizes around 100 to 1000
µ
m and a ring-shaped distribution of normal size
θ
particles. The formation of abnormal
θ
particles resulted from metal plastic flow during welding and the high content of Cu in AA2219. No abnormal
θ
particles were observed in joints of another AA2219 plate, which had a lower Cu content of 5.83 wt pct.
Laser welding of the new nickel-based alloy K439B applied for engine turbine was carried out. The effects of post-welding heat treatment (PWHT) on the microstructure and mechanical properties of the ...joints were investigated. Semi-quantitative statistical results show that after PWHT, γ’ phases in the whole joint grow up, and the radii of the primary and secondary γ’ phases are about 25 and 150 nm, respectively. The results of mechanical properties show that PWHT improves the microhardness in the weld by about HV 100, and the critical resolved shear stress provided by γ’ phases increases from 4.8 to 92 MPa. The tensile strength and yield strength of the heat-treated joint are comparable to those of the base metal. The presence of a large amount of carbides is the reason for the preferential fracture of the base metal during the tensile process.
Critical limb ischemia (CLI) is the most severe manifestation of peripheral artery disease, which is common but rarely diagnosed. Noninvasive biomarkers are urgently required to assist in the ...diagnosis of CLI. Accumulating evidence indicates that miRNAs play an important role in the development of various diseases. In this study, microarray profiling revealed 11 miRNAs with significantly altered expression in four T2DM patients with CLI compared with that in four sex- and age-matched T2DM patients without CLI. In independent cohorts, qRT-PCR validation confirmed the increased miRNA-4739 level in patients with CLI versus patients without CLI. miRNA-4739 levels increased with FPG and HbA1c (all P < 0.05). After adjusting for the risk factors, miRNA-4739 levels were found to be associated with an increased odds ratio (OR) of T2DM with CLI (OR =12.818, 95% confidence intervals (CI) 1.148 to 143.143, P = 0.038). ROC curve analysis revealed that the area under the curve (AUC) of miR-4739+confounding risk factors was 0.94 (95% CI 0.891 to 0.998, P < 0.001), which was higher than that of confounding risk factors (AUC 0.94 vs. 0.91, 95% CI -0.122 to 0.060, P > 0.05) and of miR-4739 (AUC 0.94 vs. 0.69, 95% CI -0.399 to -0.101, P < 0.001), respectively. We conclude that elevated plasma miRNA-4739 levels are independently associated with CLI in T2DM patients. miRNA-4739 is implicated as a novel diagnostic marker and a potential therapeutic target for CLI in diabetes.
Salt stress has become a major threat to plant growth. Arbuscular mycorrhizal fungi (AMF) are considered as bioameliorators and can modulate plant growth in various ways. However, few studies have ...addressed whether AMF could mediate the detrimental effects of salinity on Ligustrum vicaryi plants. In this study, inoculated (colonization with two AMF, Rhizophagus intraradices (Ri) and Funneliformis mosseae (Fm), alone and in combination) and non-inoculated (NM) L. vicaryi plants were subjected to three levels of NaCl salinity (0, 50 and 200 mM NaCl) to examine the interactive effects on the growth, chlorophyll fluorescence parameters and biochemical parameters of plants in a greenhouse experiment. The results showed that salinity stress had significanlty decreased the total dry weight of L. vicaryi plants. However, with Ri inoculation, the total dry weight, Fv/Fm, Yield, qP, the nutrient contents of P, K+, Mg2+, Zn2+, the K+:Na+ ratio and soluble protein content were considerably increased, while tissue Na+ content and shoot:root Na+ ratio were decreaed under saline conditions. It is suggested that mycorrhizal fungi can alleviate the detrimental effects of salt stress on L. vicaryi plants. Furthermore, the results also provide evidence that the mediation of N, Ca2+, Zn2+, Mg2+ and soluble protein content would be probably the main mechanism underlying salt tolerance in mycorrhizal plants.
•Arbuscular mycorrhizal fungi can mediate the detrimental effects of salinity on L. vicaryi plants.•The enhancement of N, Ca2+, Zn2+, Mg2+ and soluble protein is the mechanisms in salt tolerance for mycorrhizal plants.
Highlights • Catheter-related blood stream infection is associated with morbidity and mortality. • Silver-impregnated catheter may prevent catheter-related blood stream infection. • This ...meta-analysis included 11 randomized controlled trials and 1 cohort study. • Silver-impregnated catheter did not reduce bacterial colonization rates. • Silver-impregnated catheter did not reduce catheter-related blood stream infection.
Critical Limb Ischemia (CLI) is common but uncommonly diagnosed. Improved recognition and early diagnostic markers for CLI are needed. Therefore, the aim of our study was to identify plasma ...biomarkers of CLI in patients with type 2 diabetes mellitus (T2DM). In this study, antibody-coated glass slide arrays were used to determine the plasma levels of 274 human cytokines in four matched cases of diabetes with and without CLI. Potential biomarkers were confirmed in an independent cohort by ELISA. After adjusting for confounding risk factors, only plasma level of Siglec-5 remained significantly associated with an increased odds ratio (OR) for diabetes with CLI by binary logistic regression analysis. Receiver operating characteristic (ROC) curve analysis revealed the optimal cut-off points for Siglec-5 was 153.1 ng/ml. After entering Siglec-5, the AUC was 0.99, which was higher than that of confounding risk factors only (AUC = 0.97, P < 0.05). Siglec-5 was expressed in plaques, but not in healthy artery wall in T2DM patients. Elevated plasma Siglec-5 was independently associated with CLI in T2DM. Plasma Siglec-5 levels are implicated as an early diagnostic marker of CLI in T2DM patients and it may become a target for the prevention or treatment of CLI in diabetes.
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