DataSheet_1_Comparative study of gait parameters of patients undergoing distal femoral resections with non-operated and healthy limbs: a meta-analysis study.docx

IntroductionGait analysis is one of the most important components of functional outcome evaluation in patients with lower-extremity tumors. Disparities between operated limbs when compared with non-operated limbs and healthy populations based on gait parameters have rarely been studied. In the present study, we attempted to analyze the gait difference and its impacts on daily life.MethodsThe gait parameters of distal femoral tumor-resected patients were collected from PubMed, CNKI, MEDLINE, Embase, Cochrane, and Google Scholar till September 30, 2022, by strictly following the inclusion and exclusion criteria. Differences between gait parameters in the operated and non-operated limbs or healthy limbs of distal femoral tumor patients were analyzed based on stance phase, swing phase, cadence, and velocity. The fixed-effects and random-effects models were used to conduct a meta-analysis.ResultsSix studies were included according to the selection criteria. There were 224 patients in total in these studies. Standard mean differences were calculated for all of our outcomes. Our results showed that there was a minimal difference in the standard mean difference of gait parameters between operated and non-operated limbs and healthy limbs.ConclusionDistal femoral tumor resections have been associated with deficient muscle function and strength and impaired gait parameters. Minimal differences in the gait parameters highlighted the advantage of distal femoral resection when replaced with a prosthesis.</p

Circadian rhythm defects upon overexpression of miR-279 family miRNAs.

<p>Power of rhythmicity was determined by subtracting the significance line from the chi-squared power. Flies were defined as rhythmic for those with power of rhythmicity > = 10. The average period only considered rhythmic flies. For average power of rhythmicity, all living flies were included with arrhythmic flies having a value of 0.</p><p>Circadian rhythm defects upon overexpression of miR-279 family miRNAs.</p

The <i>mir-279/996</i> locus and phenotypes of single and double deletion alleles.

<p>(A) Annotation of the <i>mir-279/996</i> region in the <i>Drosophila</i> R5/dm3 version. RNA-Seq data (in blue) reveals characteristic "dropoffs" in read density at the Drosha cleavage sites for <i>mir-279</i> and <i>mir-996</i>. CAGE data (in black) reveals a single peak in the <i>mir-279/996</i> region located downstream of a TATA box, likely representing the start of a shared primary <i>mir-279/996</i> transcript (i.e., <i>CR31044</i>). The genomic regions deleted in five <i>mir-279/996</i> alleles are shown in red. Zoom-in of the 5' breakpoint of the <i>mir-279[ex117]</i> deletion shows it removes sequence just downstream of the <i>CR31044</i> TSS, leaving the putative TATA box intact. The mature sequences of three members of the miR-279 seed family are shown; note that miR-286 is encoded elsewhere in the genome but is more related to miR-279 than is miR-996. (B) Alignments of <i>pri-mir-279</i> and <i>pri-mir-996</i> demonstrate their distinct mature miRNAs are perfectly conserved across across 12 Drosophilids. (C) Distinct lifespan of various <i>mir-279/996</i> deletion alleles. n = 100 for <i>mir-996[ex310]</i> mutants, n>150 were assayed for the different <i>mir-279</i> mutant alleles; equal numbers of males and females were included. Error bars represent SEM. (D-H) MARCM analysis of GR21+ neurons CO₂-sensing neurons. The tester stock genotype is shown, where "X" refers to the mutations as labeled. (D, D') The normal projection pattern of GR21+ neurons to ventral glomeruli in control clones. (E, E') Deletion of <i>mir-996</i> does not perturb CO₂-sensing neurons. (F, F') <i>mir-279[ex117]</i> clones exhibit substantial ectopic projections to medial glomeruli, whereas <i>mir-279[ex36]</i> clones (G, G') and <i>mir-279/996[ex15C]</i> clones (H, H') exhibit more extensive misprojections.</p

Severe loss of mature miR-996 expression in <i>mir-279</i> deletion alleles.

<p>(A) Northern blots of miR-279 and miR-996 in various <i>mir-279</i> and <i>mir-996</i> homozygous or trans-heterozygous allele combinations. In <i>mir-279</i> alleles <i>[ex117]</i> and <i>[ex36]</i> that retain the <i>mir-996</i> genomic DNA, the levels of mature miR-996 are strongly diminished (<i>[ex117]</i>) or nearly undetectable (<i>[ex36]</i>). <i>mir-996[ex310]</i> is a deletion of the <i>mir-996</i> region that does not affect <i>mir-279</i> and <i>mir-279/996[ex15C]</i> deletes both miRNAs. (B) Quantifications of mature miR-279 and miR-996 levels. Homozygous <i>[ex117]</i> mutants expressed ~10% of the wild type level of miR-996 and <i>[ex117/ex36]</i> transheterozygous mutants expressed ~5% of miR-996. Note that the expression level for both miRNAs is copy-number dependent, since heterozygous <i>mir-279/996[ex15C]</i> flies expressed roughly half of both miRNAs compared to wild type animals.</p

Both miR-279 and miR-996 contribute to circadian rhythm.

<p>Power of rhythmicity was determined by subtracting the significance line from the chi-squared power. Flies were defined as rhythmic for those with power of rhythmicity > = 10. The average period only considered rhythmic flies. For average power of rhythmicity, all living flies were included with arrhythmic flies having a value of 0.</p><p>Both miR-279 and miR-996 contribute to circadian rhythm.</p

Modified genomic transgenes to assess individual miR-279/996 functions.

<p>(A) In the 16.6kb <i>mir-279</i>/<i>996</i> rescue transgene, the 5' end extends to cover a portion of the upstream <i>CG14508</i> gene and 3' end extends into the downstream <i>Ef1gamma</i> gene. Green and blue triangles represent <i>mir-279</i> and <i>mir-996</i> hairpins, respectively. The wildtype genomic fragment was modified to replace the <i>mir-279</i> and <i>mir-996</i> hairpins with either a deletion or the non-cognate miRNA. (B) Northern blots verify the expression of miR-279 and miR-996 from different genomic transgenes that were introduced into <i>mir-279/996[ex15C]</i> double mutant homozygous animals. RNA samples were extracted from whole flies carrying one copy of individual transgenes. Intensities of miR-279 and miR-996 expression were quantified by normalization to homozygous wild type and marked below each lane. <i>mir-996-2x</i> expressed only comparable amount of miR-996 as <i>mir-279/996-wt</i> and <i>mir-996-1x</i> transgenes. (C) Rescue of the lifespan defect in <i>mir-279/996[ex15C]</i> double mutants by 16.6kb transgenes. The <i>mir-279/996[ex15C]</i> data shown here are the same as plotted in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005245#pgen.1005245.g001" target="_blank">Fig 1C</a>. For each genotype, 100 flies including equal number of males and females were assayed; error bars represent SEM.</p

Size-Dependent Immunochromatographic Assay with Quantum Dot Nanobeads for Sensitive and Quantitative Detection of Ochratoxin A in Corn

Fluorescent microspheres are a novel luminescent nanomaterial proposed as an alternative probe to improve the detection sensitivity of competitive immunochromatographic assay (ICA). Quantum dot nanobeads (QBs) possess strong luminescence and resistance to matrix interference. Theoretically, large-sized QBs exhibit stronger luminescent intensity than small-sized QBs and are beneficial to ICA sensitivity. However, oversized QBs may reduce the sensitivity of competitive ICA. Thus, the relationship between the size and luminescent intensity of QBs and the competitive ICA sensitivity must be elucidated. In this study, QBs of different sizes (58, 124, 255, 365, and 598 nm) were synthesized. Ochratoxin A (OTA) was selected as the model analyte for competitive ICA. The effects of QB size on the detection performance of competitive ICA were then evaluated. The cutoff limit of QB-ICA for naked eye detection was used for qualitative analysis, and the half-maximal inhibitory concentration (IC₅₀) and LOD were employed for quantitative analysis. Results indicated that 124 nm QBs used as labeling probes for competitive ICA showed the optimal detection performance and the lowest cutoff value of 5 ng/mL for qualitative detection and IC₅₀ (0.39 ng/mL) for quantitative detection. Similar to commercial ELISA, QB₁₂₄-ICA displayed good accuracy, specificity, reproducibility, and practicability. In summary, 124 nm QBs can be used as a new labeling probe for competitive ICA

Comparison of gain-of-function activities of miR-279 and miR-996.

<p>(A) Luciferase sensor assays in S2 cells indicated that 3' UTRs of multiple miR-279 targets are all additionally responsive to miR-996. The control <i>Hairless</i> 3' UTR has no miR-279/996 seed match and was not repressed by these miRNAs. Error bars represent standard deviation from quadruplicate assays. (B) Northern confirmation of ectopic miR-279 and miR-996 in S2 cell experiments. pre = pre-miRNA hairpin, mature = mature miRNA product. Overexpressed miRNAs were calculated relative to endogenous mature miRNAs, normalized to 2S loading control. (C-D) Averaged activity profiles for control and miRNA overexpressing flies for 7 days in constant darkness since the second day after transferring from LD to DD. Some of these experiments utilized the amplifier driver <i>tim-UAS-Gal4</i>, as schematized in (E). (C) Overexpression of miR-279 by <i>tim-Gal4</i> induced strong arrhythmia. (D) Ectopic expression of miR-996 by <i>tim-Gal4</i> had no significant affect on circadian behavior, but further induction by <i>tim-UAS-Gal4</i> led to a complete arrhythmia. n = ~32 for each genotype; the number of flies assayed for each genotype is indicated in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005245#pgen.1005245.t002" target="_blank">Table 2</a>. (F) Validation that higher levels of mature miR-996 are induced by <i>tim-UAS-Gal4</i>, compared to <i>tim-Gal4</i>. Overexpressed miRNAs were quantified as in (B) using Northern blotting, and normalized to 2S loading control. These tests also confirm that <i>tim-Gal4</i>><i>UAS-mir-996</i> flies effectively misexpressed miR-996, even though they lacked circadian defects.</p

Both miR-279 and miR-996 contribute to maintenance of circadian rhythm.

<p>(A-F) Typical activity profiles of individual flies of various <i>mir-279/996</i> genotypes. Animals were entrained in 12hr-light/12hr-dark (LD) cycles for four days, and then kept in constant darkness (DD) for seven more days. In LD cycles, white bars represent the light phase (day) and black bars represent the dark phase (night). During DD cycles, grey and black bars represent the subjective day and night time, respectively. (A) <i>mir-279[ex117]</i> heterozygotes behave normally in that they maintain circadian activity in the dark, although the strong morning and evening activity peaks and mid-day siesta are not as well maintained. (B-C) In <i>mir-279[ex117]</i> homozygotes, the majority of animals gradually lost behavioral rhythmicity after transferring to constant darkness (B), but about 1/3 of animals could maintain circadian activity in constant darkness (C). Note that all <i>mir-279[ex117/ex117]</i> animals exhibited generally less activity than heterozygotes. The activity and circadian defects in <i>mir-279[ex117/ex117]</i> animals were rescued by single copies of the wild-type 16.6kb <i>mir-279/996</i> transgene (D) or the <i>2x-mir-279</i>-only (E) or <i>2x-mir-996</i>-only (F) transgenes. (G-J) Averaged activity profiles of various <i>mir-279/996</i> genotypes. (G) <i>mir-279/996[ex15C]</i> heterozygotes exhibit robust behavioral rhythmicity after transferring to constant darkness, but <i>mir-279[ex117]</i> homozygotes do not. In the <i>mir-279/996[ex15C]</i> homozygous background (which is normally mostly lethal by ~4 days), expression of only a single <i>1x-mir-279</i> (I) or single <i>1x-mir-996</i> (J) transgene can restore normal rhythmic behavior in constant darkness. n = ~32 for each genotype; the number of flies assayed for each genotype are indicated in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005245#pgen.1005245.t001" target="_blank">Table 1</a>.</p

Identification of two recurrent mutations of COL1A1 gene in Chinese Van der Hoeve syndrome patients

<p><b>Conclusion</b>: The two discovered mutations in <i>COL1A1</i> gene, although first reported in China, are recurrent ones that have also been found elsewhere in type I osteogenesis imperfecta patients, suggesting their role in pathogenesis of Van der Hoeve syndrome. <b>Objectives</b>: The aim of this study is to find mutational patterns of <i>COL1A1</i> gene that may account for the putative Van der Hoeve syndrome in the patients carrying symptoms of osteogenesis imperfecta, blue sclera, and conductive deafness. <b>Method</b>: Genomic DNA was extracted from the blood of each patient and exons of <i>COL1A1</i> gene were amplified using PCR and sequenced. <b>Results</b>: Sequencing in some of the two family members revealed point mutations in exon 26 (c.1792C > T) and exon 43 (c.3076C > T) of <i>COL1A1</i> gene, respectively.</p