EVALUATION OF PRE-ABLATION NLR AND LMR AS PREDICTORS OF DISTANT METASTASES IN PATIENTS WITH DIFFERENTIATED THYROID CANCER.

Objective: This research aim was to evaluates the role of the pre-ablation neutrophil-to-lymphocyte ratio (NLR) and lymphocyte-to-monocyte ratio (LMR) as predictors of distant metastases in patients with differentiated thyroid cancer (DTC). Methods: A retrospective analysis was given to 140 patients with DTC who received 131I remnant ablation after surgery. The patients were divided into two groups based on the existence of distant metastasis. Results: The two groups showed no significant difference in age, gender, WBCs, neutrophils, monocytes, eosinophils, basophils and whether the tumor was multifocal. In the univariate analysis, significant differences were found in tumor size (p=0.021), lymphocyte (p=0.012), NLR (p=0.027), and LMR (p=0.007). According to the ROC curves, NLR had an AUC of 0.612 ± 0.097 with a cut-off value of 1.845, sensitivity of 60.0%, and specificity of 66.2% (p=0.027). LMR had an AUC of 0.638 ± 0.095 with a cut-off value of 4.630, sensitivity of 84.6%, and specificity of 35.4% (p=0.007). In the multivariate analysis, larger tumor size (OR=5.246, 95% CI 1.269-10.907, p=0.009) and higher NLR (OR=2.087, 95% CI 0.977-4.459, p=0.034) were statistically significant for distant metastases. Conclusion: This research reveals that pre-ablation NLR and tumor size are significantly statistically correlated with distant metastases in patients with DTC.

ScRNA-Seq Reveals a Distinct Osteogenic Progenitor of Alveolar Bone.

The metabolism and remodeling of alveolar bone are the most active among the whole skeletal system, which is related to the biological characteristics and heterogeneity of the bone mesenchymal stromal cells (MSCs). However, there is a lack of systematic description of the heterogeneity of MSC-derived osteoblastic lineage cells as well as their distinct osteogenic differentiation trajectory of alveolar bone. In this study, we constructed a single-cell atlas of the mouse alveolar bone cells through single-cell RNA sequencing (scRNA-seq). Remarkably, by comparing the cell compositions between the alveolar bone and long bone, we uncovered a previously undescribed cell population that exhibits a high expression of protocadherin Fat4 (Fat4+ cells) and is specifically enriched around alveolar bone marrow cavities. ScRNA-seq analysis indicated that Fat4+ cells may initiate a distinct osteogenic differentiation trajectory in the alveolar bone. By isolating and cultivating Fat4+ cells in vitro, we demonstrated that they possess colony-forming, osteogenic, and adipogenic capabilities. Moreover, FAT4 knockdown could significantly inhibit the osteogenic differentiation of alveolar bone MSCs. Furthermore, we revealed that the Fat4+ cells exhibit a core transcriptional signature consisting of several key transcription factors, such as SOX6, which are involved in osteogenesis, and further demonstrated that SOX6 is required for the efficient osteogenic differentiation of the Fat4+ cells. Collectively, our high-resolution single-cell atlas of the alveolar bone reveals a distinct osteogenic progenitor that may contribute to the unique physiological characteristics of alveolar bone.

Osteoblastic RAR Inhibition Causes VAD-Like Craniofacial Skeletal Deformity.

Retinoid signaling disorders cause craniofacial deformity, among which infants with maternal vitamin A deficiency (VAD) exhibited malformation of the eye, nose, palate, and parietal and jaw bone. Previous research uncovered the pathogenesis of eye defect and cleft palate of VAD in mice, but the studies on craniofacial skeletal deformity met obstacles, and the cell/lineage and underlying mechanism remain unclear. The retinoic acid receptor (RAR) is the key transcription factor in retinoid signaling, but individual knockout cannot simulate pathway inhibition. Here, we conditionally expressed dominant-negative RARα mutation (dnRARα) in osteoblasts to specifically inhibit the transcription activity of RAR in mice, which mimics the craniofacial deformities caused by VAD in clinical cases: hypomineralization of cranial bones, mandibular deformity, and clavicular hypoplasia. Furthermore, we performed 3-dimensional reconstruction based on micro-computed tomography and confirmed the abnormalities in the shape, size, and ossification of craniofacial bones due to osteoblastic RAR inhibition. Histological analysis indicated that inhibition of RAR in osteoblasts impaired both bone formation and bone resorption, which was confirmed by transcriptome sequencing of the calvaria. Furthermore, mechanism investigation showed that inhibition of RAR in osteoblasts directly decreased osteoblast differentiation in a cell-autonomous manner by impairing osteogenic gene transcription and also inhibited osteoclast differentiation via osteoblast-osteoclast crosstalk by impairing Rankl transcription. In summary, osteoblastic RAR activity is critical to craniofacial skeletal development, and its dysfunction leads to skeletal deformities mimicking VAD craniofacial defects, providing a new insight for VAD pathogenesis.

Analysis of psychological characteristics of obese children.

OBJECTIVE: We conducted this study to analyze the psychological characteristics of obese children. PATIENTS AND METHODS: We selected 60 cases of obese children as obesity group and according to 1:1 matching principle, we selected 60 normal weight children as the control group. We investigated and analyzed children's family behavior, mental health, temperament, self-consciousness and social adaptability. RESULTS: The proportion of children with adverse behavior in the obesity group was significantly higher than that in the control group (p < 0.05). In the psychological health assessment, we compared the emotional disorder, social adjustment disorder, bad habits and behavior disorder score of both groups, and the differences were statistically significant (p < 0.05). We compared the temperament dimension score of both groups in avoidance, emotional nature, distractibility and threshold of reaction; the differences were statistically significant (p < 0.05). The proportion of negative temperament types in the obesity group were significantly higher than that in the control group (p < 0.05). We compared the self-awareness levels of both groups with regards to body appearance and properties such as gregariousness, happiness, satisfaction and total scores of self-concept aspects; the differences were statistically significant (p < 0.05). The level of social adaptive ability of the obesity group was significantly lower than that of the control group (p < 0.05). CONCLUSIONS: Children that demonstrate bad family behavior and that have a temperament which makes them difficult to raise are important factors related to obesity. Obese children often have mental and behavioral disorders, aversion, high emotional nature, low distractibility and threshold of reaction, damaged self-awareness, low self-evaluation, are not gregarious, demonstrate unhappiness and satisfaction and have poor social adaptation ability. Obesity is a cause for social concern. We need to strengthen the mental health education of obesity and promote the healthy development of children both physically and mentally.

Characterization of Dicrocoelium dendriticum isolates from small ruminants in Shaanxi Province, north-western China, using internal transcribed spacers of nuclear ribosomal DNA.

The genetic variations in internal transcribed spacers (ITS) spanning ITS-1, 5.8S and ITS-2 rDNA of Dicrocoelium dendriticum, isolated from sheep and goats in four geographical regions in Shaanxi province, were examined. The lengths of ITS-1, 5.8S and ITS-2 rDNA sequences for D. dendriticum were 749 bp, 161 bp and 234 bp, respectively. Intra-specific sequence variations of D. dendriticum were 0-0.5% for ITS-1 and 0-1.3% for ITS-2 rDNA, while the inter-specific variations among species in genus Dicrocoelium in ITS-2 rDNA were 3.4-12.3%. Phylogenetic analysis based on sequences of ITS-2 rDNA showed that all D. dendriticum isolates in the present study were grouped with reference D. dendriticum isolates from sheep and goats, and D. dendriticum isolates from cattle and Japanese serow were clustered in a sister clade. However, the phylogenetic tree could not reveal geographically genetic relationships of D. dendriticum isolates in different origins and hosts. These findings provided basic information for further study of molecular epidemiology and control of D. dendriticum infection in Shaanxi province as well as in the world.

Molecular approaches to differentiate three species of Nematodirus in sheep and goats from China based on internal transcribed spacer rDNA sequences.

Internal transcribed spacer (ITS) rDNA sequences of three Nematodirus species from naturally infected goats or sheep in two endemic provinces of China were analysed to establish an effective molecular approach to differentiate Nematodirus species in small ruminants. The respective intra-specific genetic variations in ITS1 and ITS2 rDNA regions were 0.3-1.8% and 0-0.4% in N. spathiger, 0-6.5% and 0-5.4% in N. helvetianus, and 0-4.4% and 0-6.1% in N. oiratianus from China. The respective intra-specific variations of ITS1 and ITS2 were 1.8-4.4% and 1.6-6.1% between N. oiratianus isolates from China and Iran, 5.7-7.1% and 6.3-8.3% between N. helvetianus samples from China and America. For N. spathiger, compared with samples from China, sequence differences in ITS1 rDNA were 0.3-2.4% in isolates from America, 0.3-2.9% in New Zealand and 2.1-2.4% in Australia. Genetic variations in ITS2 rDNA of N. spathiger were 0-0.4% between samples from China and America, and 0-0.8% between samples from China and New Zealand. Using mutation sites, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and specific PCR techniques were developed to differentiate these three Nematodirus species. The specific PCR assay allowed the accurate identification of N. oiratianus from other common nematodes with a sensitivity of 0.69 pg and further examination of Nematodirus samples demonstrated the reliability of these two molecular methods.

Insights into interphase large-scale chromatin structure from analysis of engineered chromosome regions.

How chromatin folds into mitotic and interphase chromosomes has remained a difficult question for many years. We have used three generations of engineered chromosome regions as a means of visualizing specific chromosome regions in live cells and cells fixed under conditions that preserve large-scale chromatin structure. Our results confirm the existence of large-scale chromatin domains and fibers formed by the folding of 10-nm and 30-nm chromatin fibers into larger, spatially distinct domains. Transcription at levels within severalfold of the levels measured for endogenous loci occur within these large-scale chromatin structures on a condensed template linearly compacted several hundred fold to 1000-fold relative to B-form DNA. However, transcriptional induction is accompanied by a severalfold decondensation of this large-scale chromatin structure that propagates hundreds of kilobases beyond the induced gene. Examination of engineered chromosome regions in mouse embryonic stem cells (ESCs) and differentiated cells suggests a surprising degree of plasticity in this large-scale chromatin structure, allowing long-range DNA interactions within the context of large-scale chromatin fibers. Recapitulation of gene-specific differences in large-scale chromatin conformation and nuclear positioning using these engineered chromosome regions will facilitate identification of cis and trans determinants of interphase chromosome architecture.

Expression of K6W-ubiquitin in lens epithelial cells leads to upregulation of a broad spectrum of molecular chaperones.

PURPOSE: Accumulation and precipitation of abnormal proteins are associated with many age-related diseases. The ubiquitin-proteasome pathway (UPP) is one of the protein quality control mechanisms that selectively degrade damaged or obsolete proteins. The other arm of the protein quality control mechanism is molecular chaperones, which bind to and help refold unfolded or misfolded proteins. We previously showed that the molecular chaperones and the UPP work in a competitive manner in eliminating the denatured proteins. To further investigate the interaction between the two protein quality control mechanisms, we determined the effects of the impairment of the UPP on the expression of molecular chaperones in human lens epithelial cells (HLEC). METHODS: K6W-ubiquitin, a dominant negative inhibitor of the UPP, was expressed in confluent HLEC via an adenoviral vector. The mRNA levels of cytoplasmic and endoplasmic reticulum (ER) chaperones were determined by real-time reverse transcription polymerase chain reaction (RT-PCR). Protein levels for these chaperones were determined by western blotting. RESULTS: Expression of K6W-ubiquitin in HLEC increased the expression of a broad spectrum of molecular chaperones. Among the heat-shock proteins, mRNA for alphaB-crystallin, Hsp70, and Hsp90 increased 27 fold, 21 fold, and twofold, respectively, in response to K6W-ubiquitin expression. Among the ER chaperones and ER stress related factors, mRNA levels of protein disulfide isomerase, Grp75, Grp78, Grp94, and the CAAT/enhancer binding protein homologous protein (CHOP) increased from 1.7 fold to 3.7 fold. The mRNA for Hsp60 also increased 1.6 fold in response to the expression of K6W-ubiquitin. The expression pattern of these chaperones in response to the expression of K6W ubiquitin is similar to that obtained when cells were treated with proteasome inhibitors or heat-shock. CONCLUSIONS: It appears that the upregulation of these chaperones is related to the elevated levels of abnormal proteins in the cells. These findings support our hypothesis that the molecular chaperones and the UPP may back each other up in the process of protein quality control. The upregulation of molecular chaperones in response to the expression of a dominant negative ubiquitin may compensate for the impairment of the UPP in the degradation of abnormal proteins.

Study on the relation between occupational fenvalerate exposure and spermatozoa DNA damage of pesticide factory workers.

AIMS: To determine sperm nuclear DNA integrity and to investigate the relation between fenvalerate (FE) exposure and spermatozoa DNA damage. METHODS: Sperm DNA fragmentation was detected by a modified alkaline single cell gel electrophoresis (Comet) assay and a terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) assay. The olive tail moment (OTM) and percentage tail DNA were measured by the Comet assay, and cell positive percentage was measured by the TUNEL assay for DNA damage evaluation. RESULTS: The DNA integrity of spermatozoa of external and internal control groups were both significantly greater than that of the FE exposed group. The median value of tail DNA percentage in the exposure group was 11.30, which was significantly higher than 5.60 in the internal control group and 5.10 in the external control group. The median value of OTM was 3.80 in the exposure group, significantly higher than 1.50 in the internal control group and 2.00 in the external control group. Mean cell positive was 31.2% in the exposure group, significantly higher than 17.4% in the internal control and 19.6% in the external control groups. Cell positive (%) was significantly correlated with tail DNA percentage and with OTM of whole subjects (n = 63). CONCLUSIONS: Results showed that occupational FE exposure is associated with an increase in sperm DNA damage. A combination of the Comet and TUNEL assays would offer more comprehensive information for a better understanding of sperm DNA damage, and the biological significance of sperm DNA damage in sperm function and male infertility.

Local structure evolution of FexNi77-xCu(1-)Nb2P14B6 soft magnetic materials by mechanical alloying.

Mechanically alloyed Fe(x)Ni77-xCu1Nb2P14B6 soft magnetic materials have been prepared with different atomic compositions. The alloy structures are investigated by X-ray absorption fine structure (XAFS). The results show that mechanical alloying (MA) can drive the Fe(x)Ni77-xCu1Nb2P14B6 powder mixture to produce amorphous alloy while the atomic concentration of Fe element is about and over 40%. On the contrary, the MA Fe(x)Ni77 xCu1Nb2P14B6 is a solid solution with a fcc-like structure in the region of lower Fe atomic concentration (<22%), preserving a medium-range order around Ni and Fe atoms. Moreover, we have found that the local structure geometry of Fe atom is similar to that of Ni atom for all the MA Fe(x)Ni77-xCu1NbP14B6 samples. It indicates that the local structures of Fe and Ni atoms in a Fe(x)Ni77-xCu1Nb2P14B6 sample only depend on the x value of element Ni after ball milling.

[Direct determination of rare earth elements in rare earth chloride and light rare earth oxide by ICP-AES].

ICP-AES was used for the direct determination of 15 rare earth elements in synthetic solutions and real sample. Spectral interferences between REEs in the mixtures of rare earth were investigated with a high-resolution echelle spectrometer and suitable analytical lines of 15 rare earth elements were selected. The Multicomponent Spectral Fitting(MSF) models were made. The method was used to remove spectral interferences and background. The factors influence the modes were discussed. The influences of acidity and ICP parameters were investigated. The compromise condition of simultaneous determination of 15 REEs was selected. Axially viewed ICP torch was used to determine 15 REEs, The detection limits are Y 0.21 microgram.L-1, La 9.1 micrograms.L-1, Ce 14.1 micrograms.L-1, Pr 1.9 micrograms.L-1, Nd 7.8 micrograms.L-1, Tm 0.37 microgram.L-1, Yb 0.12, Lu microgram.L-1, Ho 0.06 microgram.L-1, Er 0.06 microgram.L-1, Tb 0.53 microgram.L-1, Sm 1.14 micrograms.L-1, Eu 0.09 microgram.L-1, Dy 0.08 microgram.L-1, Gd 0.30 microgram.L-1. The recoveries of this procedure are between 98.4% and 101.7%. The RSD is within 2%. The method is rapid and accuracy.

Capability of serum to convert streptomycin to cytotoxin in patients with aminoglycoside-induced hearing loss.

Individual variations in sensitivity to the ototoxic effects of aminoglycoside antibiotics are well documented. Our research demonstrates that there is an apparent difference in serum from patients who are resistant or susceptible to aminoglycoside ototoxicity. In the first study, the cytotoxicity of sera from patients with and without hearing loss after various time periods following the discontinuation of aminoglycoside treatment was assayed using the isolated outer hair cell toxicity assay. The results indicate that sera from patients with hearing loss were significantly more toxic than sera from patients with normal hearing or minimal hearing loss. This toxicity may persist for up to 1 year after discontinuation of aminoglycoside therapy. In a second study, sera were obtained from patients who had received aminoglycoside therapy several years previously. None of these sera was toxic to isolated outer hair cells in vitro. Streptomycin was then incubated with the sera or a protein fraction isolated from sera, and the incubation mixtures were tested for toxicity. The percentage of damaged outer hair cells was significantly higher when streptomycin had been treated with sera or a serum protein fraction from patients with hearing loss (58+/-10% and 68+/-9%, respectively) than with sera or a serum protein fraction from a control group (10+/-5% and 17+/-4%, respectively). In addition, several incubation mixtures were analyzed using high performance liquid chromatography. A new chromatographic peak was only found in the incubations of streptomycin with serum protein from patients with hearing loss. The results suggest that sera from individuals sensitive to aminoglycoside antibiotics may metabolize these drugs to cytotoxins.