Rasal1 regulates calcium dependent neuronal maturation by modifying microtubule dynamics.

BACKGROUND: Rasal1 is a Ras GTPase-activating protein which contains C2 domains necessary for dynamic membrane association following intracellular calcium elevation. Membrane-bound Rasal1 inactivates Ras signaling through its RasGAP activity, and through such mechanisms has been implicated in regulating various cellular functions in the context of tumors. Although highly expressed in the brain, the contribution of Rasal1 to neuronal development and function has yet to be explored. RESULTS: We examined the contributions of Rasal1 to neuronal development in primary culture of hippocampal neurons through modulation of Rasal1 expression using molecular tools. Fixed and live cell imaging demonstrate diffuse expression of Rasal1 throughout the cell soma, dendrites and axon which localizes to the neuronal plasma membrane in response to intracellular calcium fluctuation. Pull-down and co-immunoprecipitation demonstrate direct interaction of Rasal1 with PKC, tubulin, and CaMKII. Consequently, Rasal1 is found to stabilize microtubules, through post-translational modification of tubulin, and accordingly inhibit dendritic outgrowth and branching. Through imaging, molecular, and electrophysiological techniques Rasal1 is shown to promote NMDA-mediated synaptic activity and CaMKII phosphorylation. CONCLUSIONS: Rasal1 functions in two separate roles in neuronal development; calcium regulated neurite outgrowth and the promotion of NMDA receptor-mediated postsynaptic events which may be mediated both by interaction with direct binding partners or calcium-dependent regulation of down-stream pathways. Importantly, the outlined molecular mechanisms of Rasal1 may contribute notably to normal neuronal development and synapse formation.

Smart Work Injury Management (SWIM) System: A Machine Learning Approach for the Prediction of Sick Leave and Rehabilitation Plan.

As occupational rehabilitation services are part of the public medical and health services in Hong Kong, work-injured workers are treated along with other patients and are not considered a high priority for occupational rehabilitation services. The idea of a work trial arrangement in the private market occurred to meet the need for a more coordinated occupational rehabilitation practice. However, there is no clear service standard in private occupational rehabilitation services nor concrete suggestions on how to offer rehabilitation plans to injured workers. Electronic Health Records (EHRs) data can provide a foundation for developing a model to improve this situation. This project aims at using a machine-learning-based approach to enhance the traditional prediction of disability duration and rehabilitation plans for work-related injury and illness. To help patients and therapists to understand the machine learning result, we also developed an interactive dashboard to visualize machine learning results. The outcome is promising. Using the variational autoencoder, our system performed better in predicting disability duration. We have around 30% improvement compared with the human prediction error. We also proposed further development to construct a better system to manage the work injury case.

[Recommendations for the diagnosis and treatment of rheumatic diseases-related hemophagocytic syndrome in China].

Hemophagocytic syndrome (HPS), which is currently named as hemophagocytic lymphohistiocytosis (HLH), is a hyperinflammatory syndrome characterized by persistent fever, hepatosplenomegaly, pancytopenia and hemophagocytosis found in bone marrow, liver, spleen and lymph nodes due to excessive activation of macrophages and cytotoxic T cells. Macrophage activation syndrome (MAS) is a specific form of HLH induced by autoinflammatory/autoimmune disorders which can be life-threatening and requires multiple disciplines. In order to improve clinicians' understanding of MAS and standardize the clinical diagnosis and treatment practice of MAS, the rheumatology branch of Chinese Rheumatology Association organized domestic experts to formulate the diagnosis and treatment standard, in order to improve the diagnosis and treatment level of MAS and improve the prognosis of patients.

[Comparison on short-term safety outcomes between off-pump and on-pump coronary artery bypass grafting by experienced surgeons: a single center study with 31 075 cases].

Objective: To compare the short-term outcomes between off-pump and on-pump coronary artery bypass graft (CABG) by experienced surgeons with similar surgical team in a single large-volume cardiac surgery center. Methods: A total of 31 075 patients with multivessel coronary disease who underwent isolated off-pump or on-pump CABG between January 1, 2009 and December 31, 2019 by experienced surgeons in Fuwai hospital were enrolled in this retrospective study. Patients was divided into on-pump CABG group and on-pump CABG group on an intention-to treat basis. Short term safety endpoints, including 30 days mortality, composite endpoint of major morbidity or mortality, prolonged postoperative length of stay (PLOS), and prolonged ICU length of stay (PICULOS), and distal anastomosis were compared between the two groups. Mortality was evaluated on 30 days post operation, other endpoints were collected before discharge. After 1∶1 propensity-score matching of baseline characteristics for on-pump and off-pump CABG, postoperative endpoints were compared with use of McNemar's test and further adjusted with the use of a logistic regression model. Results: After propensity-score matching, 10 243 matched pairs of patients were included in the final analysis, there were 4 605(22.5%) females and mean age was (60.7±8.6) years. The standardized differences were less than 5% for all baseline variables in matched cohort. Univariate analysis indicated lower risk of 30 days mortality (0.2% vs. 0.7%, P<0.001), major morbidity or mortality (5.7% vs. 8.8%, P<0.001), PLOS (3.2% vs. 4.9%, P<0.001), PICULOS (9.4% vs. 12.2, P<0.001), and lower number of distal anastomosis ((3.3±0.8) vs. (3.6±0.8), P<0.001) in off-pump CABG group than in on-pump CABG group. After adjustment of cofounders, multivariate analysis showed that off-pump CABG was still associated with a lower risk of 30 days mortality (OR=0.29, 95%CI: 0.09-0.87, P=0.027), composite endpoint of major morbidity or mortality (OR=0.60, 95%CI: 0.53-0.68, P<0.001), PLOS (OR=0.64, 95%CI 0.54-0.75, P<0.001), PICULOS (OR=0.76, 95%CI: 0.69-0.84, P<0.001). Conclusions: Off-pump CABG is related with superior short-term safety outcomes than on-pump CABG by experienced surgeons in our center.

Smart Work Injury Management (SWIM) System: Artificial Intelligence in Work Disability Management.

PURPOSE: This paper aims to illustrate an example of how to set up a work injury database: the Smart Work Injury Management (SWIM) system. It is a secure and centralized cloud platform containing a set of management tools for data storage, data analytics, and machine learning. It employs artificial intelligence to perform in-depth analysis via text-mining techniques in order to extract both dynamic and static data from work injury case files. When it is fully developed, this system can provide a more accurate prediction model for cost of work injuries. It can also predict return-to-work (RTW) trajectory and provide advice on medical care and RTW interventions to all RTW stakeholders. The project will comprise three stages. Stage one: to identify human factors in terms of both facilitators and barriers RTW through face-to-face interviews and focus group discussions with different RTW stakeholders in order to collect opinions related to facilitators, barriers, and essential interventions for RTW of injured workers; Stage two: to develop a machine learning model which employs artificial intelligence to perform in-depth analysis. The technologies used will include: 1. Text-mining techniques including English and Chinese work segmentation as well as N-Gram to extract both dynamic and static data from free-style text as well as sociodemographic information from work injury case files; 2. Principle component/independent component analysis to identify features of significant relationships with RTW outcomes or combine raw features into new features; 3. A machine learning model that combines Variational Autoencoder, Long and Short Term Memory, and Neural Turning Machines. Stage two will also include the development of an interactive dashboard and website to query the trained machine learning model. Stage three: to field test the SWIM system. CONCLUSION: SWIM ia secure and centralized cloud platform containing a set of management tools for data storage, data analytics, and machine learning. When it is fully developed, SWIM can provide a more accurate prediction model for the cost of work injuries and advice on medical care and RTW interventions to all RTW stakeholders. ETHICS: The project has been approved by the Ethics Committee for Human Subjects at the Hong Kong Polytechnic University and is funded by the Innovation and Technology Commission (Grant # ITS/249/18FX).

Secondary acute angle closure attack as an initial presentation of a novel type of systemic lupus erythematosus.

BACKGROUND: We report a rare case of secondary acute angle closure attack because of lupus choroidopathy and accompanying polyserositis, as an initial presentation of a novel type of systemic lupus erythematosus in a 44-year-old woman. CASE PRESENTATION: The patient complained of eyelid oedema and chemosis with bilateral severe loss of visual acuity. Systemic lupus erythematosus was diagnosed based on malar rash, polyserositis, proteinuria and positive antibody titers for antinuclear antibodies, anti-DNA, antinucleosome antibodies and ribosomal RNP. Subsequently, secondary bilateral acute angle closure caused by choroidal effusions with lupus choroidopathy was diagnosed. A month after steroid and immunosuppressive drug therapy, the patient's intraocular pressure and visual acuity returned to normal. During the subsequent year, the secondary acute angle closure did not recur and polyserositis remained under control. CONCLUSIONS: Bilateral, secondary acute angle closure attack due to SLE choroidopathy can be an initial presentation of SLE, which is often accompanied by polyserositis. Prompt and aggressive high doses of steroids and immunosuppressive therapy are strongly recommended.

MiR-378 promotes the cell proliferation of non-small cell lung cancer by inhibiting FOXG1.

OBJECTIVE: To identify the functioning mode of miR-378 on non-small cell lung cancer (NSCLC) and provide therapeutic targets for NSCLC. PATIENTS AND METHODS: Expression levels of miR-378 in human NSCLC tissue samples and NSCLC-derived cell lines were measured by using quantitative Real-time polymerase chain reaction (PCR). Cell proliferation capacity was assessed by methyl thiazolyl tetrazolium (MTT) assay and colony formation assay. Cell apoptosis and cell cycle distribution were identified by flow cytometry. Downstream target gene was confirmed by using luciferase and Western blotting assays. RESULTS: MiR-378 was significantly elevated in NSCLC tissues when compared with para-carcinoma tissues (n=42). Decreased-miR-378 could attenuate cell proliferation capacity, as well as promoted cell apoptosis and induced cell cycle arrest at G0/G1 phase. FOXG1 was chosen as the target gene of miR-378 by bioinformatics analysis and luciferase reporter assay. Moreover, restoration of miR-378 could impair the tumor suppression role of downregulated-miR-378 on NSCLC growth. CONCLUSIONS: Decreased-miR-378 exerted tumor-suppressive effects on NSCLC growth via targeting FOXG1 in vitro, which provided an innovative and candidate target for diagnosis and treatment of NSCLC.

Carvacrol promotes neuroprotection in the mouse hemiparkinsonian model.

Carvacrol is a monoterpene that has been linked to neuroprotection in several animal models of neurodegeneration, including ischemia, epilepsy and traumatic neuronal injury. In this study, we investigated the effects of carvacrol (i.p.) upon the neurodegeneration induced by 6-hydroxy-dopamine unilateral intrastriatal injections in mice. We have also used the cylinder test to assess the behavioral effects of carvacrol in that model of Parkinson's disease, and immunoblots to evaluate the levels of caspase-3 and TRPM7, one of major targets of carvacrol. Behavioral testing revealed that carvacrol largely reduced the asymmetrical use of the forelimbs induced by unilateral 6-hydroxy-dopamine. Carvacrol dramatically reduced the loss of tyrosine hydroxylase immunostaining both in the substantia nigra and in the striatum that are typical of the model. Immunoblots for tyrosine hydroxylase confirmed this effect. Caspase-3 levels were very high after toxin injections, but carvacrol appeared to reduce them to control levels. Finally, TRPM7, observed by immunoblots, increased after 6-hydroxy-dopamine, suggesting the involvement of this cation channel in the ensuing neurodegenerative process. The present data suggest that carvacrol promotes a marked neuroprotection in the 6-hydroxy-dopamine model of Parkinson's disease, possibly by its non-specific blocking effect upon TRPM7 channels.

[Novel biomarkers for latent tuberculosis infection by plasma proteomic profiling].

OBJECTIVE: To screen specific biomarkers for latent tuberculosis infection by comparing the plasma proteomic profiling between latent tuberculosis infection and healthy controls. METHODS: The plasma proteins from 15 cases with latent tuberculosis infection and 15 healthy controls were detected by the label-free quantitative proteomic technology. Differential expressed proteins were analyzed by GO, KEGG, and BiNGO analysis. Student's t test was used to analyze the differential expression between 2 groups. RESULTS: Twenty-three candidate proteins were identified, among which 15 proteins were downregulated (<0.5-fold at P<0.05) and 8 proteins were upregulated (>2.0-fold at P<0.05) in the latent tuberculosis infection group. Bioinformatic analysis revealed 3 proteins AAT, C3 and C4A to be the most significant. CONCLUSION: There were differential plasma protein profiles between latent tuberculosis infection and healthy controls. Candidate proteins AAT, C3 and C4A were promising biomarkers for discriminating cases with latent tuberculosis infection from healthy persons.

Targeting TYRO3 inhibits epithelial-mesenchymal transition and increases drug sensitivity in colon cancer.

Colon cancer is the third leading cause of death from cancer worldwide with less than 10% survival rate at the late stage. Although mutations of certain genes have been implicated in familial colon cancer development, the etiology of the majority of colon cancer remains unknown. Herein, we identified TYRO3 as a potential oncogene. Immunohistochemical staining results demonstrated that levels of TYRO3 were markedly elevated in polyps and colon cancer cells and were negatively correlated with prognosis. Overexpression of TYRO3 enhanced cell motility, invasion, anchorage-independent growth and metastatic ability, while knockdown of TYRO3 impaired all these processes. Results from meta-analysis showed that TYRO3 was associated with epithelial-mesenchymal transition (EMT) signatures. Gain-of-function and loss-of-function experiments demonstrated that expression of SNAI1, the master regulator of EMT, was regulated by TYRO3 and played a major role in mediating TYRO3-induced EMT processes. The murine model also demonstrated that Tyro3 and Snai1 were upregulated in the early stage of colon cancer development. To provide a proof-of-concept that TYRO3 is a druggable target in colon cancer therapy, we raised anti-TYRO3 human antibodies and showed that treatment with the human antibody abolished TYRO3-induced EMT process. More importantly, administration of this anti-TYRO3 antibody increased drug sensitivity in primary cultured colon cancer cells and xenografted mouse tumors. These findings demonstrate that TYRO3 is a novel oncogene and a druggable target in colon cancer.

Competition of infectious hypodermal and haematopoietic necrosis virus (IHHNV) with white spot syndrome virus (WSSV) for binding to shrimp cellular membrane.

Infectious hypodermal and haematopoietic necrosis virus (IHHNV) and white spot syndrome virus (WSSV) are two widespread shrimp viruses. The interference of IHHNV on WSSV was the first reported case of viral interference that involved crustacean viruses and has been subsequently confirmed. However, the mechanisms underlying the induction of WSSV resistance through IHHNV infection are practically unknown. In this study, the interference mechanisms between IHHNV and WSSV were studied using a competitive ELISA. The binding of WSSV and IHHNV to cellular membrane of Litopenaeus vannamei was examined. The results suggested that there existed a mutual competition between IHHNV and WSSV for binding to receptors present on cellular membrane of L. vannamei and that the inhibitory effects of WSSV towards IHHNV were more distinct than those of IHHNV towards WSSV.

Modified cytospin slide microscopy method for rapid diagnosis of smear-negative pulmonary tuberculosis.

OBJECTIVE: To evaluate a modified method of the Ziehl-Neelsen stain and determine whether it improves the detection rate of acid-fast bacilli (AFB) in bronchoalveolar lavage fluid (BALF) specimens. DESIGN: Bronchoscopy of patients with suspected smear-negative pulmonary tuberculosis (PTB) patients was conducted to collect BALF to assess the efficacy and accuracy of the modified method for PTB diagnosis. RESULTS: A total of 106 BALF specimens was collected from 74 PTB patients on the basis of BALF samples that were culture-positive for Mycobacterium tuberculosis. When analysed by patient, the sensitivity and specificity of our modified method were respectively 87.8% and 99.6%, while the positive predictive (PPV) and negative predictive values (NPV) were respectively 98.5% and 96.8%. Conversely, the sensitivity of direct smears and concentrated smears was respectively 16.2% and 37.8%, with 100% specificity. On analysing 106 samples, the culture positivity rate of the direct smear and the concentrated smear methods was respectively 76.4%, 13.2% and 34%, while it was 91.5% for the modified method. CONCLUSION: The sensitivity of our modified method was significantly higher than that of direct or concentrated smears. Overall, the modified method improved the detection rate of AFB in BALF specimens, and provided an efficient and accurate diagnosis of PTB in patients with suspected smear-negative PTB.

Sp1-mediated ectopic expression of T-cell lymphoma invasion and metastasis 2 in hepatocellular carcinoma.

T-cell lymphoma invasion and metastasis 2 (TIAM2) is a neuron-specific protein that has been found ectopically expressed in hepatocellular carcinoma (HCC). Results from clinical specimens and cellular and animal models have shown that the short form of TIAM2 (TIAM2S) functions as an oncogene in the tumorigenesis of liver cancer. However, the regulation of TIAM2S ectopic expression in HCC cells remains largely unknown. This study aimed to identify the mechanism underlying the ectopic expression of TIAM2S in liver cancer cells. In this report, we provide evidence illustrating that Sp1 binds directly to the GC box located in the TIAM2S core promoter. We further demonstrated that overexpression of Sp1 in HepaRG cells promotes endogenous TIAM2S mRNA and protein expressions, and knockdown of Sp1 in 2 HCC cell lines, HepG2 and PLC/PRF/5, led to a substantial reduction in TIAM2S mRNA and protein in these cells. Of 60 paired HCC samples, 70% showed a significant increase (from 1.1- to 3.6-fold) in Sp1 protein expression in the tumor cells. The elevated Sp1 expression was highly correlated with both TIAM2S mRNA and protein expressions in these samples. Together, these results illustrate that Sp1 positively controls TIAM2S transcription and that Sp1-mediated transcriptional activation is essential for TIAM2S ectopic expression in liver cancer cells.

Applicability of risk scores for postoperative nausea and vomiting in a Taiwanese population undergoing general anaesthesia.

Five popular scoring systems for postoperative nausea and vomiting (PONV) were validated and compared with two new predictive models in a Taiwanese population. Nine hundred and ninety-two patients receiving general anaesthesia in a tertiary hospital were investigated in a prospective observational cohort study. Patient demographic data and the incidence of nausea or vomiting in the first 24 hours after surgery were recorded. The overall incidence of PONV was 42%. The area under the curve (AUC) of the five published PONV risk scoring systems was 0.62 to 0.67. Logistic regression analysis in this study cohort showed that female sex and a history of PONV/car sickness were the only statistically significant independent risk factors for PONV (likelihood ratio test P <0.001).The AUCs of our two-predictor and gender-only models were 0.668 and 0.643, respectively (Nagelkerke R² = 0.122 and 0.109). Goodness-of-fit showed that a two-predictor model predicted outcome that was in agreement with the observed outcome (P=0.973). Both the two-predictor model and the Apfel score had a similar AUC that was significantly different from the AUCs of the other models. The AUC for the gender-only model in our population was similar to that of the simplified Koivuranta and the Palazzo and Evans scores (AUC=0.659 and 0.632; P=0.137 and 0.513 respectively). All AUCs had only moderate discrimination power but our female gender-only model was much simpler. Using female gender as the only predictor of PONV had predictive power with 75% sensitivity and 54% specificity.

Clinical and genetic analysis of Taiwanese patients with hereditary spastic paraplegia type 5.

BACKGROUND AND PURPOSE: Spastic paraplegia type 5 (SPG5) is an autosomal recessive (AR) hereditary spastic paraplegia (HSP) associated with pure or complicated phenotypes. This study aimed to screen SPG5 in Taiwanese HSP patients. METHODS: Sequencing of the SPG5 gene, CYP7B1, was performed in a cohort of 25 ethnic Han Taiwanese patients with AR or sporadic HSP. Clinical information and magnetic resonance imaging (MRI) were analyzed in confirmed SPG5 patients. RESULTS: One (33%) AR kindred and four (18%) sporadic cases had CYP7B1 mutations. All of the SPG5 cases carried the mutation c.334 C>T (R112X). Haplotype analysis suggested a 'founder effect' in ethnic Hans for this mutation. The phenotype was either pure or complicated by cerebellar ataxia. For the primary HSP phenotype, there were profound dorsal column sensory deficits in all patients. Spine MRI showed thoraco-lumbar cord atrophy in some patients. CONCLUSIONS: Spastic paraplegia type 5 is a common cause of AR and sporadic HSPs that has a higher frequency in Taiwanese than in other ethnic groups. It is associated with a CYP7B1 founder mutation and its phenotype is characterized by pronounced dorsal column sensory loss, with cerebellar ataxia in some patients.

A miniature lighted stylet for fast oral endotracheal intubation in rabbits.

Efficient oral endotracheal intubation of laboratory animals is a challenging technique in veterinary research. This study introduces a miniaturized lighted stylet for rabbit intubation. An experiment with repeated measures on two factors was used to assess the feasibility and efficacy of this method. The first factor compared stylet intubation vs. laryngoscopic intubation. The second compared three practitioners, one with prior experience and two without. Success rates on the initial attempt were not statistically different (χ(2)=2.46, P=0.12). The time difference between methods was significant (F=41.007, P<0.001), although the effect of practitioners was not (F=1.038, P=0.365). The mean±SD of the intubation time, combining results from the three practitioners, was 20.34±17.15s for the stylet method and 57.58±64.21s for the laryngoscopic method. The results of this study demonstrate that lighted stylet intubation is efficient, robust, and independent of practitioner experience.

Novel exon nucleotide substitution at the splice junction causes a neonatal Marfan syndrome.

The fibrillin-1 gene (FBN1) mutations are associated with a broad spectrum of disorders including Marfan syndrome (MFS) and show great clinical heterogeneity. An underrepresentation for mutations leading to premature termination codon (PTC) in FBN1 exons 24-32 was found in neonatal or severe MFS but the underlying cause was unclear. This study thoroughly examined two FBN1 mutations on exons 24-32 region to illustrate the molecular mechanisms underlying these FBN1 mutations on MFS etiology. Two nucleotide substitutions, c.3208G> C, the last nucleotide of exon 26, and c.3209A>G, the first nucleotide of exon 27, affecting the same amino acid, p.D1070H and p.D1070G, respectively, gave very different phenotypes. We demonstrate that c.3208G>C generates two alternatively spliced transcripts, while c.3209A>G does not affect the splicing. We further demonstrate that the aberrantly spliced transcripts do not go through nonsense-mediated decay, but rather produce unstable, premature protein peptides that are degraded by endoplasmic reticulum associated degradation. The molecular mechanism outlined here defines a model for the pathogenesis of PTC-containing mutation within the exons 24-32 of FBN1 in MFS. Furthermore, our data suggest that PTC mutation within this region may lead to early lethality in neonatal MFS.

The effects of a fat- and sugar-enriched diet and chronic stress on nonalcoholic fatty liver disease in male Wistar rats.

PURPOSE: The pathogenesis of nonalcoholic fatty liver disease (NAFLD) is still under debate. The aim of this study was to investigate the effects of a long-term fat- and sugar-enriched diet (FSED) and chronic stress (CS) on NAFLD. METHODS: Male Wistar rats were fed on either a standard diet or a FSED and given CS, a random electric foot shock (2 hr/morning and afternoon per day), or not for 12 weeks. After the experimental period, epididymal adipose tissue weight, sign of visceral obesity (VO), and hepatic index (HI) were measured. At sacrifice blood samples and liver were obtained. Histology of the liver was blindly determined by a pathologist. RESULTS: Histopathologically, moderate to severe steatosis, ballooning hepatocytes, and portal or lobules inflammation were observed in the FSED+CS group. However, mild to moderate steatosis with a few portal inflammation in the FSED group and mild steatosis or not with a few portal inflammation in the CS group were found correspondingly. In addition, more severe blood-fat disorder, high HI, fatty metabolic dysfunction, oxidative stress, high expressions of C-reactive protein mRNA and low expressions of peroxisome proliferator-activated receptor alpha mRNA in the liver were also revealed in the FSED+CS group. But, the degree of VO was not different between the FSED and FSED+CS groups. CONCLUSION: The observations strongly suggest that chronic stress can aggravate fat- and sugar-enriched diet-induced NAFLD from steatosis to steatohepatitis in male Wistar rats, although VO is not changed.