Evaluation of circulating miRNAs and mRNAs expression patterns in autism spectrum disorder

Background Autism spectrum disorder is a condition related to brain development that affects a person’s perception and socialization, resulting in problems in social interaction and communication. It has no single known cause, yet several different genes appear to be involved in autism. As a genetically complex disease, dysregulation of miRNA expression and miRNA–mRNA interactions might be a feature of autism spectrum disorder. The aim of the current study was to investigate the expression profile of circulating miRNA-128, miRNA-7 and SHANK gene family in ASD patients and to assess the possible influence of miRNA-128 and miRNA-7 on SHANK genes, which might provide an insight into the pathogenic mechanisms of ASD and introduce noninvasive molecular biomarkers for the disease diagnosis and prognosis. Quantitative real-time PCR technique was employed to determine expression levels of miRNA-128, miRNA-7 and SHANK gene family in blood samples of 40 autistic cases along with 30 age- and sex-matched normal volunteer subjects. Results Our study revealed a statistical significant upregulation of miRNA-128 expression levels in ASD cases compared to controls (p value < 0.001). A statistical significant difference in SHANK-3 expression was encountered on comparing cases to controls (p value < 0.001). However, miRNA-7 expression showed no significant difference between the studied groups. Conclusions MiRNA-128 and SHANK-3 gene are emerging players in the field of ASD. They are promising candidates as noninvasive biomarkers in autism. Future studies are needed to emphasize their pivotal role.

Elevated Levels of PD-L1 on MDSCs in Patients with Ph(-) Myeloproliferative Neoplasm

Introduction. We previously reported that PD-1 and PD-L1 were increased in patients with Ph(-) myeloproliferative neoplasm (MPN) ( Wang et al., Leuk Res 2019). The PD-1 inhibitor therapy or immune checkpoint inhibitor therapy (ICI) trial in MPN by Hobbs et al. reported a negative result (Blood, 2020 (supplement )). Resistance to ICI therapy has been reported to be related to myeloid-derived suppressor cells (MDSCs) in melanoma and breast carcinoma. We also previously reported that MDSCs were increased in patients with Ph(-) MPN (Wang et al., Leu Res 2016). Regulation of immune suppression by MDSCs has been reported to be related to PD-1and PD-L1 expression on the MDSC. Therefore, the current study measured PD-1 and PD-L1 expression in MDSCs in patients with Ph(-) MPN. Materials and Methods. Twenty-six patients, including 11 essential thrombocythemias (ETs), six polycythemia vera (PV), and nine myelofibrosis (six primary MF, one post-ET-MF, two post-PVMF) were compared with ten normal volunteer controls. MDSCs were measured as follows: the pelleted PBMNCs were used for Magnetic-assisted cell separation (MACS, Miltenyi Biotech, Inc.). PBMNCs are sequentially selected for HLA-DR - cells, from which we further selected for CD14 + and CD14 - cells using Miltenyic magnetic microbeads kits, respectively, according to the manufacturer's protocol. The resulting HLA-DR -CD14 + and HLA-DR -CD14 - cells were stored at -80°C until use. Flow cytometric assay:HLA-DR -CD14 + and HLA-DR -CD14 - cells were stained with both anti-human CD274 (PD-L1) FITC and human CD279 (PD-1) PE (BD Biosceinces, Inc.), together with anti-human CD14 APC or anti-human CD33 APC (BD Biosceinces, Inc.), respectively. Flow cytometric assessment was done using the BD Accuri C6 flow cytometer. While HLA-DR -CD14 +Monocytic MDCS cells were gated and assayed for surface expression of PD-1 (CD274) and PDL-1 (CD279), G-MDSC (granulocytic MDSC), and M-NDSC (Monocytic MDSC) were assayed as HLA-DR -CD14 - , CD33 + and HLA-DR -CD14 +, respectively. These MDSCs were then assayed for surface expression of PD-1 (CD274) and PDL-1 (CD279). The flow data were analyzed using FlowJo V8 (Flowjo, LLC), and the mean fluorescent intensity (MFI) was calculated. Results. PD-L1 on the m-MDSCs was significantly elevated in patients with MPN (grouping patients with ET, PV, and MF) and MF than controls. PD-1 on the M-MDSCs was not different between ET, PV, MF, or MPN compared with normal controls. Compared to controls, PD-L1 on the G-MDSC was significantly elevated in patients when ET, PV, and MF were grouped as MPN. PD-1 on the G-MDSC were not different between ET, PV, MF, or MPN and controls. Conclusion. Ph(-) MPN was found to have significantly elevated PD-L1 on the M-MDSCs and G-MDSCs, but PD-1 levels were not significantly elevated. Further studies to employ drugs, such as a combination of IMiDs (e.g., lenalidomide) and anti-MDSC drugs in combination with ICI in vitro and clinical trials, may be warranted. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

Clinical severity of serum S100A12 levels in patients with osteoarthritis and S100A12 open inflammation of osteoarthritis model by NLRP3

Osteoarthritis is a common chronic bone and joint disease, which is characterized by degenerative changes and destruction of articular cartilage, secondary hyperostosis. This study aimed to investigate the clinical severity and mechanism of S100A12 in patients with osteoarthritis. Serum samples were obtained from patients with osteoarthritis or normal volunteer in Minhang Branch of Yueyang Hospital of Integrated Traditional Chinese and Western Medicine affiliated to Shanghai University of Traditional Chinese Medicine (Shanghai, China). C57BL/6J mice performed Resection of the medial collateral ligament and medial meniscus as mice model. MC3T3-E1 cells were induced with 100 ng of LPS as vitro model. The serum level of S100A12 was increased in patients with osteoarthritis. Similarly, S100A12 levels of serum and bone tissue from mice model of osteoarthritis were also higher than those of sham group. Over-expression of S100A12 promoted inflammation levels while down-regulation of S100A12 decreased inflammation levels in in vitro model of osteoarthritis. NLRP3 is an important target of S100A12 in pro-inflammation effects of osteoarthritis. NLRP3 was involved in the effects of S100A12 on inflammation in in vitro model of osteoarthritis. S100A12 also accelerated inflammation by NLRP3 in mice model of osteoarthritis. We conclude that serum S100A12 levels was a possible clinical severity, open inflammation of osteoarthritis model by NLRP3 and its receptors may be effective in preventing the development of osteoarthritis.

PERBEDAAN OPTIC NERVE SHEATH DIAMETER (ONSD) PADA RESPONDEN NORMAL DAN NYERI KEPALA

Nyeri kepala merupakan penanda adanya gangguan intrakranial. Nervus optikus merupakan saraf kranialis yang mudah dipengaruhi langsung adanya gangguan intrakranial. Kalimatnya Jumping. Kelainan nervus optikus dapat diketahui dengan adanya pelebaran optic nerve sheath diameter (ONSD). ONSD dapat diukur dengan transorbital sonografi yang bersifat nonimvasif. Penelitian ini bertujuan membedakan diameter ONSD pada orang yang nyeri kepala dan orang normal. Penelitian ini menggunakan pendekatan observasional analitik dengan metode cross sectional. Responden diambil dari pasien rawat inap dan responden normal. Hasil penelitian didapatkan rata – rata nilai ONSD pada pasien nyeri kepala adalah 0,52 ± 0,86 dan pada responden normal adalah 0,40 ± 0,57 (p < 0,05). Terdapat perbedaan signifikan nilai ONSD pada pasien normal dan nyeri kepala. Penelitian ini membuktikan bahwa terdapat gangguan intrakranial pada orang dengan nyeri kepala dan dapat digunakan sebagai deteksi dini.Kata Kunci: Optic Nerve Sheath Diameter (ONSD), Responden Normal, Nyeri KepalaHeadache is a sign of intracranial disorders. The optic nerve is a cranial nerve that is easily affected directly by intracranial disorders. The sentence is Jumping. Optic nerve abnormalities can be identified by the widening of the optic nerve sheath diameter (ONSD). ONSD can be measured by transorbital sonography which is nonimvasive. This study aims to distinguish the ONSD diameter in people with headaches and normal people. This research uses an analytic observational approach with cross sectional method. Respondents were taken from inpatients and normal respondents. The results showed that the average value of ONSD in headache patients was 0.52 ± 0.86 and in normal respondents was 0.40 ± 0.57 (p <0.05). There is a significant difference in the value of ONSD in normal patients and headaches. This research proves that there are intracranial disorders in people with headaches and can be used as early detection.Keyword: Optic nerve sheath diameter , headache , normal volunteer

Myoelectric Activity of Individual Lumbar Erector Spinae Muscles Variation by Differing Seat Pan Depth

Background: The chair influences the position of the user in relation to his or her devices. Prolonged static sitting is a frequently mentioned risk factor for low back pain. Seat design, thus, plays an important role in the study of human sitting. Quantitative information is needed on what happens to body when one sits in chairs with different seat depth. Objective: To determine the myoelectric activity (EMG) of individual lumbar erector spinae muscles after sitting in chairs with different seat pan depth. Methods: EMG recordings were taken using surface electrodes placed on the lumbar erector spine muscles of 25 normal, volunteer subjects. EMG recordings for muscle activity were made while the study participants were in a comfortable position and performed the required tasks. The experiments investigated with 3 seat depths according to the 5th, 50th and 95th percentiles of the buttock popliteal length. The recorded EMG data were normalized to the maximal voluntary contraction. The mean EMG recording was calculated for each of the 3 chairs tested. A mixed model was used to assess the differences among the situations. Results: A significant (p<0.05) difference was observed between the mean EMG recordings for the 3 tested seat pan depths. EMG activity was higher in seats with the 5th and 95th percentiles compared with that for the seat with 50th percentile of buttock popliteal length depth. Conclusion: The seat pan depth used during a comfortable position has a significant effect on the level of myoelectric activity in the lumbar erector spinal muscles. The finding of this study may contribute to our understanding of the biomechanics of sitting.

VALIDATION OF THE FINNISH VERSION OF THE MONTREAL COGNITIVE ASSESSMENT TEST1

The aim of this study was to determine the clinical utility of the Finnish version of the MoCA test for screening Alzheimer’s disease and MCI. The purpose was to examine the ability (sensitivity and specificity) of the MoCA to distinguish patients with AD and MCI from cognitively normal controls. The study population consists of three participant groups: patients with AD (n=25), patients meeting the criteria for MCI (n=18), and cognitively normal controls (NC) (n=39). The AD group consists of subjects with very mild (CDR= 0.5, n=12), mild (CDR=1, n=12), and moderate (CDR=2, n=1) dementia, and they were given a diagnosis of dementia by using the revised NINCDS-ARDRA criteria. The normal control group (NC) consists of 39 cognitively normal volunteer participants. The three study groups differed from each other in terms of sex, age, and level of education. The NCs were younger than the subjects with AD (t [37,374] = 3.265, p = 0.002) and MCI (t [30,800] = 4.306, p = < 0.001). The NCs were also better-educated than the patients with AD (t [54,975] = -3.419, p = 0.001) and MCI (t [40,782] = -3.008, p = 0.004). The sensitivity and specificity of the MoCA in detecting AD and MCI was done according to various cutoff points. With a cutoff score of 26, the MoCA had a sensitivity of 100% to detect subjects with AD and a sensitivity of 100% to detect subjects with MCI. The specificity was 79.5%. With a cutoff score of 24, which was the best threshold in the present study, the MoCA not only had a high sensitivity to detect subjects with AD (96%) and MCI (89%) but also delivered a high specificity (97%). The MoCA has a high sensitivity and specificity to detect subjects with AD and MCI with a cutoff score of 24/30. The Finnish version of the MoCA is a feasible screening instrument for assessing cognitive decline. According to our study, the optimal cutoff score of the MoCA is 24/30.

Programmed Cell Death Receptor (PD-1), PD-1 Ligand (PD-L1) Expression in Philadelphia Chromosome Negative Myeloid Neoplasms

Introduction : PD-1 and PD-L1 pathway inhibitor therapy has been extensively explored in the field of oncology treatment recently, however there is no information so far about its usefulness in myeloid neoplasms (MPN). We have previously reported on PD-1 and PD-L1 expression in Philadelphia chromosome negative myeloid neoplasm (MPN), but now that we have accumulated more cases , we wish to report the final results. Materials and Methods: 63 patients with Philadelphia chromosome negative MPN in total were studied which included 16 MF (14 PMF, 2 post-PV-MF, 2 post-ET-MF), 29 ET, 18 PV, and 25 normal volunteer controls. Flow Cytometry Studies: Leukocytes were obtained from whole blood after RBC lysis using RBC lysis buffer (Qiagen). To quantify surface bound PD-1 and PDL-1, the leukocytes were incubated with CD279-APC and CD274-PE, as well as CD4-FITC, CD8-FITC, CD14-PCP, or CD34-FITC (BD Biosciences) and were subjected to flow cytometric analysis. Results: PD-1 and PD-L1 positive cells were expressed as a percentage in gated CD4bright, CD8+, CD14+, or CD34+ cells. We found that 1) there was no difference in PD-1 or PD-L1 levels among different groups of MPN, but there was a significant difference when they were grouped of PV, ET and MF as MPN and compared with controls in all the immune cells including CD4+, CD8+, CD14 + and,CD34+ cells.(Representative of CD4+ shown inFig(1a). 2) High expression levels (more than 50% cell positivity) was found on the CD34+ cells with PD-1 and PD-L1 expression of 20% and 26% in MPN patients respectively ( Fig 1b,1c) . 3) We also found that there was no correlation between of the PD-1 or , PD-L1 expression levels with clinical features including such as WBC counts , platelet counts, and hemoglobin levels, , or presence or absence of the JAK2 , MPL, or CALR gene mutations or splenomegaly Conclusion: We have shown elevated increased expression levels of PD1 and PDL-1 on the immune cells and high expression ( more than 50%) in the CD34+ cells , suggest that PD-1 or , PD-L1 pathway inhibitor therapy maybe worthwhile in Philadelphia chromosome negative Ph(-) MPN .. Disclosures Wang: celgene: Other: clinical trial.

Dynamic assessment of the wrist after total wrist arthroplasty

Our aim was to assess the outcome in patients with total wrist arthroplasty performed for end stage wrist osteoarthritis. We analysed the ranges of motion of operated and un-operated wrists using a flexible electrogoniometer during the Sollerman hand function test. We assessed grip strength with a digital dynamometer and completed patient reported outcome scores more than one year post-operatively. We reviewed 12 patients with a mean age of 64 (range 48–82) years. The flexion-extension arc was 72% and radioulnar deviation arc was 53% of the un-operated side but the total range of motion (area of circumduction) was 43% of the un-operated side and only 20% of the circumduction in age and gender-matched normal volunteers. Peak grip strength was 68% of the un-operated side. The patients reported good outcome with mean Michigan Hand Questionnaire (MHQ) scores of 56 (range 25–84) and mean Patient Evaluation Measure (PEM) scores of 39 (range 20–68). Patients completed the activities of Sollerman hand function test in twice the time (6 min) as required for a normal volunteer (2.8 min). The circumduction ellipses were narrow and central with limited radio-ulnar deviation and small mean areas of motion during activities of daily living.