Adalimumab Dose Reduction and Withdrawal in Stable Non-Infectious Pediatric Uveitis: An Open-Label, Prospective, Pilot Study.

PURPOSES: This study investigated the feasibility of adalimumab (ADA) dose reduction and withdrawal strategy in children with stable pediatric non-infectious uveitis (PNIU). METHODS: This open-label prospective pilot trial recruited 18 stable PNIU patients (33 eyes) between two and eighteen years old who were treated with standard doses of ADA (20/40 mg every 2 weeks) plus oral methotrexate. The interval of ADA injection was extended to 4 weeks and followed up for 24 weeks. If the uveitis remained stable, ADA was discontinued and followed up for another 24 weeks. ADA was considered successfully stopped if no relapse occurred during this period. The relapse-free survival rate, best corrected visual acuity (BVCA), anterior chamber cell (ACC), vitritis, macular thickness (MT), and serum ADA levels were evaluated. Approval Number: 2021KYPJ201. ClinicalTrials.gov identifier: NCT05155592. RESULTS: The relapse-free survival rate was 22.2% (4/18) at 48 weeks. 33.3% (6/18) of patients relapsed when ADA was given every 4 weeks, while 44.5% of patients (8/18) relapsed after ADA was stopped. The four patients successfully withdrawn from ADA were all diagnosed with BD. No statistically significant differences (p > 0.05) were observed in BCVA and MT between baseline and final follow-up. The proportion of ACC and vitritis exhibited an upward trend (p < 0.05) during follow-up. Serum ADA gradually decreased to zero during follow-up in both non-recurrence and recurrence groups. CONCLUSIONS: In PNIU children who reached remission for 6 months, ADA dose reduction and withdrawal were associated with a high risk of inflammation recurrence. Timely adjustment of ADA to the last effective dosage frequency can regain control of the inflammation. Detection of ADA serum levels in patients with recurrence may help find the appropriate interval of ADA use.

Connecting cellular mechanisms and extracellular vesicle cargo in traumatic brain injury.

Traumatic brain injury is followed by a cascade of dynamic and complex events occurring at the cellular level. These events include: diffuse axonal injury, neuronal cell death, blood-brain barrier break down, glial activation and neuroinflammation, edema, ischemia, vascular injury, energy failure, and peripheral immune cell infiltration. The timing of these events post injury has been linked to injury severity and functional outcome. Extracellular vesicles are membrane bound secretory vesicles that contain markers and cargo pertaining to their cell of origin and can cross the blood-brain barrier. These qualities make extracellular vesicles intriguing candidates for a liquid biopsy into the pathophysiologic changes occurring at the cellular level post traumatic brain injury. Herein, we review the most commonly reported cargo changes in extracellular vesicles from clinical traumatic brain injury samples. We then use knowledge from animal and in vitro models to help infer what these changes may indicate regrading cellular responses post traumatic brain injury. Future research should prioritize labeling extracellular vesicles with markers for distinct cell types across a range of timepoints post traumatic brain injury.

Comparison of clinical outcomes between posterolateral approach and modified Hardinge approach in salvage total hip arthroplasty after medial buttress plate failure for femoral neck fractures.

OBJECTIVE: The purpose of this study was to investigate the clinical outcomes of salvage total hip arthroplasty (THA) after medial buttress plate surgery for femoral neck fractures via the modified Hardinge approach (MHA) and posterolateral approach (PLA) through a retrospective analysis. PATIENTS AND METHODS: From October 2016 to October 2020, a total of 41 patients with failed femoral neck fractures treated with cannulated screws and medial buttress plates underwent unilateral salvage THA, and a retrospective study was conducted. According to the surgical approach, patients were divided into PLA group and MHA group. Clinical and radiological data were evaluated. The primary outcome indicators were the Pain Visual Analog Scale (VAS) and Hip Harris Score (HHS). Secondary outcome indicators include hemoglobin (HGB), hematocrit (HCT), creatine kinase (CK), creatine kinase-MB (CK-MB), etc. The occurrence of postoperative complications was also recorded. RESULTS: There were no differences in demographic or clinical characteristics before surgery. There were no differences in postoperative HGB, HCT, CK-MB and radiological parameters. The surgical approach had no effect on the hospitalization period. The PLA group had earlier ambulation time, and the serum level of CK was also low. Analysis of the HHS and VAS showed that on postoperative day 3, the PLA group had superior scores. The incidence of complications did not significantly differ between groups. CONCLUSIONS: The posterolateral approach for salvage THA provides better functional recovery with less muscle damage in the early postoperative period.

Association between serum asprosin and diabetic peripheral neuropathy in patients with type 2 diabetes mellitus in the community.

OBJECTIVE: This study aimed to investigate the relationship between serum asprosin level and diabetic peripheral neuropathy (DPN) in community patients with type 2 diabetes mellitus (T2DM). PATIENTS AND METHODS: A total of 498 patients with T2DM were recruited from Zhuoma Community Health Service Station and Chengbei West Street Community Health Service Center in Changzhi City of Shanxi Province between November 2019 and July 2021. Their height, weight, and body mass index (BMI), as well as fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c), triglyceride (TG), and serum asprosin levels, were analyzed. Patients were divided into the DPN group (n = 329) and the non-DPN group (n = 169) according to the presence or absence of DPN. The t-test, Mann-Whitney U test, and χ² test were used to compare the indicators between the two groups. Pearson or Spearman correlation analysis was used to evaluate the correlation between serum asprosin and other clinical data. Multivariate logistic regression analysis was used to analyze the influencing factors of DPN. RESULTS: Compared with the non-DPN group, the DPN group had higher serum asprosin (p < 0.05). The prevalence of DPN gradually increased according to the tertiles of asprosin (56%, 67%, and 75%; p < 0.05). Multivariate logistic regression analysis showed that after adjustment for covariates, patients with asprosin concentrations between 295.4-367.0 pg/ml and concentrations > 367.0 pg/ml had a higher risk of diabetic neuropathy compared than those with asprosin levels < 295.4 pg/ml (p < 0.05). CONCLUSIONS: Serum asprosin was found to be positively correlated with DPN, and it resulted as an influencing factor for DPN in patients with T2DM in the community. With the increase of asprosin, the risk of DPN also increased.

Predicting Drug Treatment Outcomes in Childrens with Tuberous Sclerosis Complex-Related Epilepsy: A Clinical Radiomics Study.

BACKGROUND AND PURPOSE: Highly predictive markers of drug treatment outcomes of tuberous sclerosis complex-related epilepsy are a key unmet clinical need. The objective of this study was to identify meaningful clinical and radiomic predictors of outcomes of epilepsy drug treatment in patients with tuberous sclerosis complex. MATERIALS AND METHODS: A total of 105 children with tuberous sclerosis complex-related epilepsy were enrolled in this retrospective study. The pretreatment baseline predictors that were used to predict drug treatment outcomes included patient demographic and clinical information, gene data, electroencephalogram data, and radiomic features that were extracted from pretreatment MR imaging scans. The Spearman correlation coefficient and least absolute shrinkage and selection operator were calculated to select the most relevant features for the drug treatment outcome to build a comprehensive model with radiomic and clinical features for clinical application. RESULTS: Four MR imaging-based radiomic features and 5 key clinical features were selected to predict the drug treatment outcome. Good discriminative performances were achieved in testing cohorts (area under the curve = 0.85, accuracy = 80.0%, sensitivity = 0.75, and specificity = 0.83) for the epilepsy drug treatment outcome. The model of radiomic and clinical features resulted in favorable calibration curves in all cohorts. CONCLUSIONS: Our results suggested that the radiomic and clinical features model may predict the epilepsy drug treatment outcome. Age of onset, infantile spasms, antiseizure medication numbers, epileptiform discharge in left parieto-occipital area of electroencephalography, and gene mutation type are the key clinical factors to predict the epilepsy drug treatment outcome. The texture and first-order statistic features are the most valuable radiomic features for predicting drug treatment outcomes.

Identification of novel Mendelian disorders of the epigenetic machinery (MDEMs)-associated functional mutations and neurodevelopmental disorders.

BACKGROUND: Mendelian disorders of the epigenetic machinery (MDEMs) are a newly identified group of neurodevelopmental disorders (NDDs) and multiple congenital anomalies caused by mutations in genes encoding components of the epigenetic machinery. Many studies have shown that MDEM-associated mutations may disrupt the balance of chromatin states and trigger dysplasia. AIM: To help eight Chinese families with NDDs acquire a definitive diagnosis. METHODS: In this study, we used whole-exome sequencing to diagnose eight unrelated Chinese families with NDDs. We also verified the potential pathogenic variants by Sanger sequencing and analyzed the changes in gene expression along with histone methylation modifications. RESULTS: Eight variants of six epigenetic machinery genes were identified, six of which were novel. Six variants were pathogenic (P) or likely pathogenic (LP), while two novel missense variants (c.5113T>C in CHD1 and c.10444C>T in KMT2D) were classified to be variants of uncertain significance (VUS). Further functional studies verified that c.5113T>C in CHD1 results in decreased protein levels and increased chromatin modifications (H3K27me3). In addition, c.10444C>T in KMT2D led to a significant decrease in mRNA transcription and chromatin modifications (H3K4me1). Based on experimental evidence, these two VUS variants could be classified as LP. CONCLUSION: This study provided a definitive diagnosis of eight families with NDDs and expanded the mutation spectrum of MDEMs, enriching the pathogenesis study of variants in epigenetic machinery genes.

[Analysis of clinical and psychological characteristics in 250 patients with fibromyalgia].

To analyze the clinical and psychological characteristics of fibromyalgia (FM), so as to further understand and improve the capability of identifying FM. The clinical data of 250 FM patients diagnosed in the outpatient clinic of the Department of Rheumatology, the First Medical Center, Chinese PLA General Hospital, from December 2019 to September 2021, were collected and analyzed. The patients aged 40 (31.0, 52.3) years, including 188 female patients (75.2%) and 62 male patients (24.8%). There was a statistically significant difference in age comparison between female [42.5 (33.0,54.0) years] and male patients [32.5 (27.8,43.5) years] (P<0.001). The score of pain degree was 6 (4, 8), and [7 (5, 8)] of female patients was higher than [6 (4, 7)] of the male patients (P=0.040). The widespread pain index (WPI) was 13 (10,15). The regions with high pain incidence were left shoulder girdle (87.2%, 218/250), right shoulder girdle (86.8%, 217/250), upper back (86.4%, 216/250), neck (79.6%, 199/250) and lower back (77.6%, 194/250) and etc. The incidence of chest pain in female patients (55.3%, 104/188) was lower than that in male patients (75.8%, 47/62) (P=0.004). The symptom severity scale (SSS) score was 8 (7-10). 74.6% (185/248) suffered from anxiety and 77.5% (193/249) suffered from depression in 249 patients. Female patients were more common in FM patients than male patients, the median age of female patients was older than that of male patients, and the median score of pain severity of female patients was higher than male patients. Shoulders girdle, upper back, neck and lower back were the most frequently reported pain regions, and the incidence of chest pain in female patients was lower than that in male patients. The incidence of major non-painful symptoms was high and the proportion with anxiety or depression was high. The above clinical features are very helpful for early diagnosis of FM.

Mechanisms and Biomarker Potential of Extracellular Vesicles in Stroke.

Stoke is a prevalent and devastating neurologic condition with limited options for therapeutic management. Since brain tissue is rarely accessible clinically, peripheral biomarkers for the central nervous system's (CNS's) cellular response to stroke may prove critical for increasing our understanding of stroke pathology and elucidating novel therapeutic targets. Extracellular vesicles (EVs) are cell-derived, membrane-enclosed vesicles secreted by all cell types within the CNS that can freely pass the blood-brain barrier (BBB) and contain unique markers and content linked to their cell of origin. These unique qualities make brain-derived EVs novel candidates for non-invasive blood-based biomarkers of both cell specificity and cell physiological state during the progression of stroke and recovery. While studies are continuously emerging that are assessing the therapeutic potential of EVs and profiling EV cargo, a vast minority of these studies link EV content to specific cell types. A better understanding of cell-specific EV release during the acute, subacute, and chronic stages of stroke is needed to further elucidate the cellular processes responsible for stroke pathophysiology. Herein, we outline what is known about EV release from distinct cell types of the CNS during stroke and the potential of these EVs as peripheral biomarkers for cellular function in the CNS during stroke.

Application of offset Dingo instruments in Anterior to Psoas (ATP)/Oblique Lumbar Interbody Fusion (OLIF) procedure: A retrospective study of 80 patients.

OBJECTIVE: Oblique lumbar interbody fusion (OLIF) is a minimally invasive lumbar fusion procedure that has flourished in recent years. However, complications related to the anterior-to-psoas (ATP) approach have been reported. The purpose of this study was to assess the OLIF technique using offset design "Dingo" instruments for the treatment of degenerative lumbar disorders. METHODS: A retrospective review was performed to identify patients between March 2018 and November 2020 who underwent OLIF with conventional or modified instruments with a minimum follow-up of 12months. Demographic data were recorded. Operation time, intraoperative blood loss, and length of hospital stay were compared between cohorts. Imaging parameters were measured and compared between cohorts before and after surgery. Clinical outcomes were assessed at each follow-up visit. All complications were reviewed. RESULTS: A total of 80 consecutive patients were included. The mean follow-up times for the conventional group and modified group were 14.5months and 16.3months, respectively. The incidence of hip flexion weakness was 27.5% and 7.5% for the conventional cohort and modified cohort, respectively. The modified group was associated with a shorter operation time, shorter length of hospital stay, higher postoperative disk height and better cage position. The symptoms of all patients improved after surgery. At 1-week follow-up, the modified group showed significantly better clinical scores. CONCLUSIONS: OLIF with modified instruments and techniques shows promising outcomes in reducing approach-related complications. Additionally, the modified technique is associated with a better cage position, shorter length of hospital stay and less postoperative pain in the early period.

[Epidemiological and etiological characteristics of Vibrio parahaemolyticus strains causing foodborne disease outbreaks in Guangdong Province from 2017 to 2020].

Objective: To study the epidemiological and pathogenic characteristics of Vibrio parahaemolyticus isolated from outbreaks cases in Guangdong Province, 2017-2020. Methods: Epidemiological characteristics of 87 outbreak events caused by Vibrio parahaemolyticus were analyzed. Strains were serotyped, and then analyzed by pulsed-field gel electrophoresis (PFGE). Results: The food-borne disease outbreak caused by Vibrio parahaemolyticus was found in 16 cities. 44.8% (39/87) and 37.9% (33/87) of the outbreaks occurred in hotels, restaurants and school canteens, respectively. Improper food processing and storage (40.2%, 35/87) and cross contamination caused by indiscriminate raw and cooked food (25.3%, 22/87) were the main causes of food-borne disease outbreaks of Vibrio parahaemolyticus. The main serotypes of patient derived strains were O3:K6 (87.5%) and O4:KUT (22.5%). The similarity value between O3:K6 type isolates was 65.5%-100.0%, and the PFGE pattern similarity value of O4:KUT type isolates was 66.5%-100.0%. Conclusion: Outbreaks caused by Vibrio parahaemolyticus are widely distributed in Guangdong province. It is necessary to strengthen the publicity and education on the correct handling of food in hotels, restaurants, schools, and unit canteens. O3:K6 and O4:KUT serotypes are the main serotypes of the outbreak. There is genetic diversity among the epidemic strains.

Cave-in decompression technique in percutaneous endoscopic transforaminal discectomy for ossification occupation in lumbar spinal canal: A retrospective analysis of 23 cases.

OBJECTIVE: Percutaneous endoscopic transforaminal discectomy (PETD) for the treatment of ossification occupation of the lumbar spinal canal is technically demanding. The purpose of this study was to describe an endoscopic cave-in decompression technique in PETD for ossification occupation of the lumbar spinal canal and to report the clinical results. METHODS: From May 2018 to June 2020, 23 consecutive cases, diagnosed in our institution as ventral ossification in the lumbar spinal canal and treated with PETD, were evaluated. The endoscopic cave-in decompression technique was performed. We analyzed the clinical outcomes on a visual analogue scale (VAS) and the Oswestry Disability Index (ODI). Perioperative data and complications rate were also recorded. RESULTS: Mean follow-up was 15.7±2.8months. Mean preoperative VAS score for low-back pain was 5.65±1.43, and decreased to 0.57±0.5 at final follow-up. Mean preoperative VAS score for leg pain was 5.48±1.38, and decreased to 0.56±0.5 at final follow-up. Mean preoperative ODI score decreased from 33.04±5.28 preoperatively to 8.7±2.54 at last follow-up. One patient experienced transient postoperative hypoesthesia, and one developed a mild transient decline in muscle strength; both recovered progressively. Postoperative CT and MRI showed that the ossification was effectively removed and the nerve root and dural mater was completely decompressed in all cases. CONCLUSION: The endoscopic cave-in decompression technique in PETD is a safe and effective treatment method for selected patients with ossification occupation of the lumbar spinal canal.

[Clinicopathological features of congenital hemangioma: a study of 40 cases].

Objective: To investigate the clinical and pathologic features, diagnosis and differential diagnosis of congenital hemangioma (CH). Methods: Forty cases of CH were diagnosed from January 2017 to December 2020 in Henan Provincial People's Hospital. The clinical and pathological and immunohistochemical data were analyzed, with review of literature. Results: There were 24 male and 16 female patients. The lesions were located in the head, neck (11 cases), limbs (14 cases), and trunk (15 cases). The clinical manifestations were congenital painless plaques or masses, the larger ones protruded on the skin surface, mostly dusky purple or bright red, with surrounding white halos. Under low magnification, the tumor was lobular and well demarcated, composed of neo-microvascular lumen of different sizes. The vascular endothelial cells were cuboidal or hobnail in appearance, forming stellar drainage vessels within the lobules. Extra-medullary hematopoiesis was seen in one case of rapidly involuting CH; there were different number of tortuous and dilated vascular lumen between the lobular structures, and some non-involuting CH cases were vascular malformations, which were devoid of lobulated structures. Immunohistochemistry showed that endothelial cells were strongly positive for CD31, CD34 and ERG, while D2-40 and GLUT-1 were negative. Conclusions: CH is a benign congenital vascular tumor with characteristic lobulated growth and abnormal blood vessels in the stroma. Pathological diagnosis often needs to be differentiated from infantile hemangioma, pyogenic granuloma, kaposiform hemangioendothelioma and vascular malformation.

Robust bulk micro-nano hierarchical copper structures possessing exceptional bactericidal efficacy.

Conventional copper (Cu) metal surfaces are well recognized for their bactericidal properties. However, their slow bacteria-killing potency has historically excluded them as a rapid bactericidal material. We report the development of a robust bulk superhydrophilic micro-nano hierarchical Cu structure that possesses exceptional bactericidal efficacy. It resulted in a 4.41 log10 reduction (>99.99%) of the deadly Staphylococcus aureus (S. aureus) bacteria within 2 min vs. a 1.49 log10 reduction (96.75%) after 240 min on common Cu surfaces. The adhered cells exhibited extensive blebbing, loss of structural integrity and leakage of vital intracellular material, demonstrating the rapid efficacy of the micro-nano Cu structure in destructing bacteria membrane integrity. The mechanism was attributed to the synergistic degradation of the cell envelope through enhanced release and therefore uptake of the cytotoxic Cu ions and the adhesion-driven mechanical strain due to its rapid ultimate superhydrophilicity (contact angle drops to 0° in 0.18 s). The scalable fabrication of this micro-nano Cu structure was enabled by integrating bespoke precursor alloy design with microstructure preconditioning for dealloying and demonstrated on 2000 mm2 Cu surfaces. This development paves the way to the practical exploitation of Cu as a low-cost antibiotic-free fast bactericidal material.

[Clinicopathological features of verrucous hemangioma].

Objective: To investigate the clinicopathological features, and differential diagnosis of verrucous hemangioma (VH). Methods: Twenty-eight VH cases diagnosed from 2005 to 2020 in Henan Provincial People's Hospital, Zhengzhou, China were analyzed retrospectively. Immunohistochemical studies were used to detect diagnostic markers. The mutation status of PIK3CA (exons 9 and 20) was detected using fluorescence PCR. Results: There were 13 males and 15 females in 28 cases, with the male to female ratio of 1.0∶1.2. There were 25 patients under the age of 18 years. The age range was from 10 months to 56 years (mean, 9.7 years; median, 4.5 years). There were 17 cases occurred in the lower extremities, 7 in the upper extremities and 4 in the trunk. All 28 cases were irregular red patches on the skin, which grew slowly. Some of them were thickened with uneven surface, which was light pink or red-white. Skin lesions of the 7 cases ranged from dark red and reddish brown, with a rough and hard surface. Satellite foci were present. Microscopically, 28 cases had a wide range of pathological features. Dilated, malformed vessels were observed from dermal papilla to deep soft tissue. Among them, the dermal papillary layer was mainly composed of many proliferating and expanding thin-walled capillaries and cavernous blood vessels. Thin-walled small vessels were found in the dermal reticular layer and subcutaneous fascia layer, with no obvious endothelial cell proliferation, occasional papillary hyperplasia, and lobular distribution of the malformed vessels in the fascia layer mixed with the fibroadipose tissue. There was epidermal papillary hyperplasia with hyperkeratosis and parakeratosis, lengthening and mutual fusion of epithelial horns. Immunohistochemistry showed that CD31, CD34, ERG and WT-1 were diffusely and strongly positive. The expression of GLUT-1 was present in superficial dermal vascular endothelial cells, but undetectable in the deep layer. The PIK3CA tests of 13 cases showed that no somatic mutations were found in exons 9 and 20. Twenty-five patients were followed up for 5 months to 10 years. Seven patients underwent multiple surgical resections and plastic surgeries due to the large size, and 8 patients had recurrence. Conclusions: VH is a rare congenital vascular malformation and more commonly occurs in infants and children. It tends to appear in limbs, especially lower limbs and distal limbs. Its morphology and immunophenotype are characteristic and should be distinguished from other vascular malformations and the resolution phase of infant hemangiomas. In about one third of the cases, postoperative recurrence may occur and long-term follow-up is often required.

[Oxymatrine improves renal fibrosis and inflammation in diabetic rats by modulating CHK1/2 phosphorylation].

OBJECTIVE: To explore the role of cell cycle checkpoint kinase 1/2 (CHK1/2) in mediating the inhibitory effect of oxymatrine (OMT) against renal inflammation and fibrosis in diabetic rats. METHODS: SD rats were randomly divided into normal control group, diabetes model group (DM) and OMT treatment group (n=6). HE and Masson staining were used to observe histopathological changes of the renal tissue, and the expressions of CHK1, CHK2, p-CHK1 and p-CHK2 were localized by immunohistochemical staining. The contents of interleukin-6 (IL-6) and IL-1ß in the renal tissue were detected using ELISA, and the expression levels of CHK1, CHK2, p-CHK1, p-CHK2, type Ⅲ collagen (Col-Ⅲ), type Ⅳ collagen (Col-Ⅳ), and fibronectin (FN) were determined using Western blotting. The changes in the expressions of CHK1, CHK2, p-CHK1, p-CHK2, Col-Ⅲ, Col-Ⅳ and FN proteins were also examined with Western blotting in NRK-52E cells in response to high glucose exposure, OMT treatment and siRNA-mediated CHK1/2 knockdown. RESULTS: In diabetic rats, OMT treatment significantly decreased the levels of blood glucose, serum creatinine and 24 h urinary protein (P < 0.05) and obviously improved inflammatory cell infiltration and fibrosis phenotype in the renal tissue (P < 0.05). CHK1 and CHK2 were mainly expressed in the cytoplasm and nuclei of renal tubule cells, and their phosphorylation levels were significantly higher in DM group than in the control group and OMT group. OMT treatment significantly decreased the protein expression levels of p-CHK1, p-CHK2, Col-Ⅲ, Col-Ⅳ and FN in the renal tissue of diabetic rats and in NRK-52E cells exposed to high glucose (P < 0.05). In NRK-52E cells, CHK1/2 knockdown resulted in significant reduction of the protein expressions of p-CHK1/2, Col-Ⅲ, Col-Ⅳ and FN (P < 0.05). CONCLUSION: The inhibitory effects of OMT against renal inflammation and fibrosis in diabetic rats are mediated probably by lowered phosphorylation levels of CHK1 and CHK2, which result in reduced release of the downstream inflammatory mediators and decreased secretion and deposition of extracellular matrix.

[MiR-671-5p negatively regulates SMAD3 to inhibit migration and invasion of osteosarcoma cells].

OBJECTIVE: To explore the role of miR-671-5p in regulating the migration and invasion of osteosarcoma and the underlying mechanisms. METHODS: The differentially expressed microRNAs (miRNAs) in osteosarcoma were screened in the NCBI online database, and the target proteins of these miRNAs were predicted and their functions were analyzed. Osteosarcoma cells were transfected with a plasmid overexpressing miR-671-5p, and the transfection efficiency was assessed using quantitative real-time PCR (qRT-PCR). The changes in the migration and invasion of the transfected cells were examined with Transwell assay, and the expressions of proteins related with epithelial-mesenchymal transition (EMT) were detected using Western blotting. Dual-luciferase reporter assay was performed to determine whether the 3'UTR of SMAD3 contained a targeted binding site of miR-671-5p. RESULTS: MiR-671-5p was significantly down-regulated in both osteosarcoma tissues and osteosarcoma cells (P < 0.05). The osteosarcoma cells overexpressing miR-671-5p showed significantly reduced migration and invasion abilities (P < 0.05) with obviously lowered expressions of EMT-related proteins (P < 0.05). SMAD3 was highly expressed in osteosarcoma cells (P < 0.05), and dual-luciferase reporter assay confirmed the presence of a targeted binding site between miR-671-5p and the 3'UTR of SMAD3 (P < 0.05). In osteosarcoma cells transfected with a SMAD3-overexpressing plasmid (P < 0.05), the high expression of SMAD3 significantly inhibited by miR-671-5p overexpression (P < 0.05). Transwell assay demonstrated that SMAD3 overexpression significantly promoted the migration and invasion of osteosarcoma cells (P < 0.05), and while miR-671-5p overexpression obviously reversed this effect (P < 0.05). CONCLUSION: MiR-671-5p can inhibit the invasion and migration of osteosarcoma cells by negatively regulating SMAD3.

Melatonin protects against excessive autophagy-induced mitochondrial and ovarian reserve function deficiency though ERK signaling pathway in Chinese hamster ovary (CHO) cells.

Excessive autophagy-induced follicular atresia of ovarian granulosa cells might be one of the pathogenesis of Premature Ovarian Insufficiency (POI), and melatonin (MT) exerted many beneficial effects on mitochondria. However, there was little report regarding the beneficial effects of MT on excessive autophagy-induced mitochondrial and ovarian reserve function deficiency, and the mechanisms have not been clearly identified. Autophagy played a protective role in cells survival, however, high level of autophagy could lead to cell death. In this report, firstly, Chinese hamster ovary cell damage model stably expressing EGFP-LC3 was established. Next, we systematically investigated the protective effects of MT on mitochondrial and ovarian reserve function and molecular mechanisms using this cell damage model. Our results revealed that 10-9 M MT not only protected against the decline of anti-mullerian hormone (AMH) expression induced by excessive autophagy, but also rescued excessive autophagy-induced impairment of mitochondrial expression and mitochondrial membrane potential. Furthermore, MT protected against excessive autophagy-induced decrease of nucleus-encoded proteins including SDHA and mitofilin, and mitochondrial dynamic-related proteins including OPA1, MFN2, and DRP1. MT also decreased mitochondrial oxidative stress, increased antioxidant enzyme superoxide dismutase 2 (SOD2) expression and ameliorated the G2/M cell cycle arrest induced by excessive autophagy. Finally, MT inhibited excessive autophagy-induced activation of extracellular signal regulated kinase (ERK) signaling pathway. In conclusion, our study showed that MT rescued impairment of mitochondrial and ovarian reserve function, and production of mitochondrial ROS and cell cycle arrest induced by excessive autophagy through down-regulated ERK pathway, implying the potential therapeutic drug target for POI.

Micromagnetic simulation of microstructure effect for binary-main-phase Nd-Ce-Fe-B magnets.

We investigate the magnetic properties of a chemically heterogeneous binary-main-phase (BMP) Nd-Ce-Fe-B magnet with a core-shell structure via micromagnetic simulation. It is found that the coercivity strongly depends on the shell thickness. The BMP magnet's coercivity initially increases and then decreases with increasing Nd-rich shell thickness, and so there is the optimal shell thickness which shows the maximum coercivity for any given Ce concentration. The simulation shows the significant difference in coercivity and maximum energy product between the BMP and single-main-phase magnets. Notably, the magnetization reversal mechanism of the BMP magnet is revealed in the simulation. Local reversals in the BMP magnet first occur in the Ce-rich shells, followed by the Nd-rich cores. Then, the magnetization in Ce-rich core/Nd-rich shell typed grains is switched after reversed magnetization of all the Nd-rich core/Ce-rich shell typed grains. The BMP magnet represents a further increased coercivity for a larger GB thickness, which can be well explained by a maximum stray field.

InterCarb: A Community Effort to Improve Interlaboratory Standardization of the Carbonate Clumped Isotope Thermometer Using Carbonate Standards.

Increased use and improved methodology of carbonate clumped isotope thermometry has greatly enhanced our ability to interrogate a suite of Earth-system processes. However, interlaboratory discrepancies in quantifying carbonate clumped isotope (Δ47) measurements persist, and their specific sources remain unclear. To address interlaboratory differences, we first provide consensus values from the clumped isotope community for four carbonate standards relative to heated and equilibrated gases with 1,819 individual analyses from 10 laboratories. Then we analyzed the four carbonate standards along with three additional standards, spanning a broad range of δ47 and Δ47 values, for a total of 5,329 analyses on 25 individual mass spectrometers from 22 different laboratories. Treating three of the materials as known standards and the other four as unknowns, we find that the use of carbonate reference materials is a robust method for standardization that yields interlaboratory discrepancies entirely consistent with intralaboratory analytical uncertainties. Carbonate reference materials, along with measurement and data processing practices described herein, provide the carbonate clumped isotope community with a robust approach to achieve interlaboratory agreement as we continue to use and improve this powerful geochemical tool. We propose that carbonate clumped isotope data normalized to the carbonate reference materials described in this publication should be reported as Δ47 (I-CDES) values for Intercarb-Carbon Dioxide Equilibrium Scale.