RNA sequencing steps toward the first line

• DNA is the sequence that codes for proteins.
• Messenger RNA is transcribed from the DNA sequence of genes and translated into protein.
• It can be difficult to predict how a change in the DNA sequence will affect messenger RNA and protein quantity and quality.
• DNA translocation changes can cause the joining of sequences from two different genes or different parts of the same gene.
• DNA sequencing is often used clinically to predict how DNA changes might affect proteins.
• Alternatively, RNA sequencing can be used as a more direct measure of the effect of DNA changes on the protein products.
• This sequencing is important for identifying changes in cancer that may indicate response to targeted therapy, prognosis, or diagnosis.

     

Fluoxetine-induced hepatic lipid accumulation is mediated by prostaglandin endoperoxide synthase 1 and is linked to elevated 15-deoxy-Δ12,14PGJ2

Major depressive disorder and other neuropsychiatric disorders are often managed with long-term use of antidepressant medication. Fluoxetine, an SSRI antidepressant, is widely used as a first-line treatment for neuropsychiatric disorders. However, fluoxetine has also been shown to increase the risk of metabolic diseases such as non-alcoholic fatty liver disease. Fluoxetine has been shown to increase hepatic lipid accumulation in vivo and in vitro. In addition, fluoxetine has been shown to alter the production of prostaglandins which have also been implicated in the development of non-alcoholic fatty liver disease. The goal of this study was to assess the effect of fluoxetine exposure on the prostaglandin biosynthetic pathway and lipid accumulation in a hepatic cell line (H4-II-E-C3 cells). Fluoxetine treatment increased mRNA expression of prostaglandin biosynthetic enzymes (Ptgs1, Ptgs2, and Ptgds), PPAR gamma (Pparg), and PPAR gamma downstream targets involved in fatty acid uptake (Cd36, Fatp2, and Fatp5) as well as production of 15-deoxy-Δ^12,14PGJ₂ a PPAR gamma ligand. The effects of fluoxetine to induce lipid accumulation were attenuated with a PTGS1 specific inhibitor (SC-560), whereas inhibition of PTGS2 had no effect. Moreover, SC-560 attenuated 15-deoxy-Δ^12,14PGJ₂ production and expression of PPAR gamma downstream target genes. Taken together these results suggest that fluoxetine-induced lipid abnormalities appear to be mediated via PTGS1 and its downstream product 15d-PGJ₂ and suggest a novel therapeutic target to prevent some of the adverse effects of fluoxetine treatment.     

Biomarkers of in vivo limbal stem cell function

PurposeTo evaluate in vivo parameters as biomarkers of limbal stem cell function and to establish an objective system that detects and stage limbal stem cell deficiency (LSCD).MethodsA total of 126 patients (172 eyes) with LSCD and 67 normal subjects (99 eyes) were included in this observational cross-sectional comparative study. Slit-lamp biomicroscopy, in vivo laser scanning confocal microscopy (IVCM), and anterior segment optical coherence tomography (AS-OCT) were performed to obtain the following: clinical score, cell morphology score, basal cell density (BCD), central corneal epithelial thickness (CET), limbal epithelial thickness (LET), total corneal nerve fiber length (CNFL), corneal nerve fiber density (CNFD), corneal nerve branch density (CNBD), and tortuosity coefficient. Their potential correlations with the severity of LSCD were investigated, and cutoff values were determined.ResultsAn increase clinical score correlated with a decrease in central cornea BCD, limbal BCD, CET, mean LET, maximum LET, CNFL, CNFD, CNBD, and tortuosity coefficient. Regression analyses showed that central cornea BCD, CET and CNFL were the best parameters to differentiate LSCD from normal eyes (Coef = 3.123, 3.379, and 2.223; all p < 0.05). The rank correlation analysis showed a similar outcome between the clinical scores and the central cornea BCD (ρ = 0.79), CET (ρ = 0.82), and CNFL (ρ = 0.71). A comprehensive LSCD grading formula based on a combination of these parameters was established.ConclusionsA comprehensive staging system combining clinical presentation, central cornea BCD, CET, and CNFL is established to accurately and objectively diagnose LSCD and stage its severity.     

红外光谱数据融合对栽培滇重楼产地鉴别*

目的 多样的环境因素使得不同产地栽培滇重楼的化学成分也丰富多样，不同居群栽培滇重楼的甾体皂苷类成分具有很大的差异，多源数据融合分析能更全面的表征药材化学信息，建立一个高效而准确的产地鉴别模型，为其资源合理开发利用提供依据。方法 以来自云南和四川的8个产地（保山、楚雄、大理、红河、丽江、成都、文山、玉溪）共366份栽培滇重楼根茎为实验材料，采集其傅里叶变换近红外光谱（FT-NIR）和衰减全反射-傅里叶变换中红外光谱（ATR-FTMIR）数据。采用Kennard-Stone算法将不同产地的样品分为2/3的训练集和1/3的预测集，基于4种特征变量提取方法（CARS、VIP、SPA、SO-Covsel）结合2种数据融合策略（低级数据融合和中级数据融合），建立偏最小二乘产地判别分析模型。根据模型参数交叉验证均方根误差（RMSECV）和预测均方根误差（RMSEP）评估模型的稳定性，模型训练集和预测集准确率（ACC）评估模型分类性能。结果 近红外光谱和中红外光谱均能反应不同产地栽培滇重楼的化学成分差异，在中级数据融合中，基于VIP和SPA提取的特征变量建立的模型正确率均大于94%。相较于中级数据融合，低级数据融合模型得到了最为满意的结果，其预测集分类正确率达到100%。结论 根据近红外和中红外数据建立的低级数据融合PLS-DA模型，能够用于栽培滇重楼的产地鉴别分析。     

Isokinetic quadriceps symmetry helps in the decision to return to running after anterior cruciate ligament reconstruction

BackgroundAfter anterior cruciate ligament reconstruction (ACLR), the decision to allow a return to running is empirical, and the post-operative delay is the most-used criterion. The Quadriceps isokinetic-strength Limb Symmetry Index (Quadriceps LSI), with a cutoff of 60%, could be a useful criterion.ObjectiveTo determine the association between a Quadriceps LSI ≥ 60% and return to running after ACLR.MethodsOver a 10-year period, we retrospectively included 470 patients who underwent ACLR. Four months after ACLR, participants performed an isokinetic test; quadriceps concentric peak torque was used to calculate the Quadriceps LSI at 60?/s. With a Quadriceps LSI ≥ 60%, a return to running was suggested. At 6 months after ACLR, participants were clinically evaluated for a return to sport and post-operative middle-term complications. A multivariable predictive model was built to assess the efficiency diagnosis of this cutoff in order to consider cofounding factors. Quadriceps LSI cutoff ≥ 60% was assessed with sensitivity, specificity and the area under the receiver operating characteristic curve (AUC).ResultsAccording to our decision-making process with the 60% Quadriceps LSI cutoff at 60?/s, 285 patients were authorized to return to running at 4 months after ACLR and 185 were not, but 21% (n = 59) and 24% (n = 45), respectively, were not compliant with the recommendation. No iterative autograft rupture or meniscus pathology occurred at 6 months of follow-up. On multivariable logistic regression analysis, a return to running by using the 60% Quadriceps LSI cutoff was associated with undergoing the hamstring strand procedure (odds ratio 2.60, 95% confidence interval [CI] 1.75–3.84; P < 0.0001) and the absence of knee complications (1.18, 1.07–1.29; P = 0.001) at 4 months. The sensitivity and specificity of the 60% Quadriceps LSI cutoff were 83% and 70%, respectively. The AUC was 0.840 (95% CI 0.803–0.877).ConclusionsUsing the 60% cutoff of the isokinetic Quadriceps LSI at 4 months after ACLR could help in the decision to allow a return to running.     

Low percentage of patients passed the ‘Back in Action’ test battery 9 months after bone-patellar tendon-bone anterior cruciate ligament reconstruction

BackgroundAnterior cruciate ligament (ACL) reconstruction is recommended in patients who intend to return to high-level sports. However, there is only a 55–80% return to pre-injury level of sports after ACL reconstruction, with a re-injury rate up to 20%. The aim of this study was to determine the percentage of patients passing the Back in Action (BIA) test 9 months after primary bone-patellar-tendon-bone (BPTB) ACL reconstruction, and evaluate the association between passing the BIA test and patient reported outcome measurements (PROMs).MethodsPatients underwent the BIA test 9 months after BPTB ACL reconstruction. In total 103 patients were included. Passing the BIA test (PASSED-group) was defined as a normal or higher score at all sub-tests with limb symmetry index (LSI) ≥90% for the dominant leg and LSI >80% for the non-dominant leg. Patients who did not meet these criteria were defined as the FAILED-group. PROMs included the International Knee Documentation Committee, Knee injury Osteoarthritis Outcome Score and Anterior Cruciate Ligament-Return to Sport after Injury.ResultsEighteen patients (17.5%) passed the BIA test 9 months after BPTB ACL reconstruction. PROMs were not statistically significant different between the PASSED- and FAILED-group.ConclusionLow percentage of patients passed the BIA test 9 months after BPTB ACL reconstruction. Although current PROMs cut-off values were met, the BIA test results show persistent functional deficits. Therefore, the BIA test could be of additional value in the decision-making process regarding return to sport (RTS). This study highlights the need for additional rehabilitation as RTS in a condition of incomplete recovery may increase the risk of re-injury.Level of evidenceII.     

Bilateral changes in knee joint laxity during the first year after non-surgically treated anterior cruciate ligament injury

ObjectivesAnalyse changes in knee laxity between 3, 6, 12 and 24 months after non-surgically treated ACL injury and to analyse associations between knee laxity and knee function, self-reported knee stability, ACL-Return to Sport after Injury (ACL-RSI), fear and confidence at different timepoints during recovery.DesignProspective cohort study.Participants125 patients (67 males, mean age 25.0 ± 7.0 years) with acute ACL injury.Main outcomeLaxity was measured using KT-1000 arthrometer. Self-reported knee function was assessed using the International Knee Documentation Committee Subjective Knee Form (IKDC-SKF). Confidence and fear were assessed with questions from the ACL-RSI scale. Subjectively knee stability was assessed using SANE.ResultsKnee laxity increased bilaterally from 3 to 12 months, and in the non-involved knee from 3 to 24 months (p˂0.05), although mean change was below 1 mm. Side-to-side difference in knee laxity was correlated with IKDC-SKF (r = −0.283) and knee stability in rehabilitation/sport activities (r = −0.315) at 6 months, but not with confidence/fear.ConclusionKnee laxity increased bilaterally during the first year after non-surgically treated ACL injury, though, the mean change in knee laxity was below 1 mm and the clinical significance is unknown. Knee laxity was weakly associated with knee function and perceived knee stability.Level of evidenceLevel IITrial registrationNCT02931084     

水痘-带状疱疹病毒性角膜葡萄膜炎继发青光眼1例报告及文献复习

分析水痘-带状疱疹病毒（VZV）性角膜葡萄膜炎继发青光眼患者的临床表现，探讨其治疗方案。