Alteration in brain functional connectivity in patients with post-stroke cognitive impairment during memory task: A fNIRS study.

OBJECTIVE: This study attempted to evaluate the functional connectivity (FC) in relevant cortex areas during three memory tasks using the functional near-infrared spectroscopy (fNIRS) method to expound the neural mechanisms in individuals with post-stroke cognitive impairment (PSCI). METHODS: Short-term memory and visuospatial abilities were assessed using the clock drawing test, digit span test, and Corsi Block-tapping tests with simultaneous fNIRS. The oxygenated hemoglobin concentration signals were recorded from the bilateral motor sense cortex (LMS/RMS) and prefrontal lobe (LPFT/PFT/RPFT) of 19 subjects with cognitive impairment (PSCI group), 27 stroke subjects (STR group) and 26 healthy subjects (HC group). RESULTS: MMSE scores were positively correlated with the clock drawing test and digit span test scores but not with Corsi Block-tapping scores. During each test, functional connectivity between the bilateral MS (LMS/RMS) was highest within each group, but the functional connectivity between motor sense cortex and frontal lobe was lowest. PSCI group showed decreased FC between bilateral motor sense cortex (P < 0.05) and between motor sense cortex and frontal lobe (P > 0.05) during clock drawing test and Corsi Block-tapping test while decreased FC between each region of interest during digit span test with no significant difference. Functional connectivity levels were closely related to MMSE scores. CONCLUSIONS: Decreased functional connectivity level may be a marker of impaired cognitive function in post-stroke cognitive impairment. The fNIRS-based functional connectivity provides a non-invasive method to recognize cognitive impairment post-stroke. Functional connectivity changes may help to further understand the neural mechanisms of cognitive impairment post stroke.

Ultrashort wave alleviates oxygen-glucose deprivation/reoxygenation injury via up-regulation of SPCA1 expression in N2a cells. / è¶ çŸ­æ³¢é€šè¿‡ä¸Šè°ƒSPCA1的表达减轻N2a细胞氧糖剥夺复氧后的损伤.

OBJECTIVES: Application of ultrashort wave (USW) to rats with cerebral ischemia and reperfusion injury could inhibit the decrease of expression of secretory pathway Ca2+-ATPase 1 (SPCA1), an important participant in Golgi stress, reduce the damage of Golgi apparatus and the apoptosis of neuronal cells, thereby alleviating cerebral ischemia-reperfusion injury. This study aims to investigate the effect of USW on oxygen-glucose deprivation/reperfusion (OGD/R) injury and the expression of SPCA1 at the cellular level. METHODS: N2a cells were randomly divided into a control (Con) group, an OGD/R group, and an USW group. The cells in the Con group were cultured without exposure to OGD. The cells in the OGD/R group were treated with OGD/R. The cells in the USW group were treated with USW after OGD/R. Cell morphology was observed under the inverted phase-contrast optical microscope, cell activity was detected by cell counting kit-8 (CCK-8), apoptosis was detected by flow cytometry, and SPCA1 expression was detected by Western blotting. RESULTS: Most of the cells in the Con group showed spindle shape with a clear outline and good adhesion. In the OGD/R group, cells were wrinkled, with blurred outline, poor adhesion, and lots of suspended dead cells appeared; compared with the OGD/R group, the cell morphology and adherence were improved, with clearer outlines and fewer dead cells in the USW group. Compared with the Con group, the OGD/R group showed decreased cell activity, increased apoptotic rate, and down-regulating SPCA1 expression with significant differences (all P<0.001); compared with the OGD/R group, the USW group showed increased cell activity, decreased apoptotic rate, and up-regulating SPCA1 expression with significant differences (P<0.01 or P<0.001). CONCLUSIONS: USW alleviates the injury of cellular OGD/R, and its protective effect may be related to its up-regulation of SPCA1 expression.

Functional near-infrared spectroscopy during motor imagery and motor execution in healthy adults. / 健康成人运动想象与运动执行期间的近红外脑功能成像.

OBJECTIVES: Studies on the influence of motor imagery (MI) on brain structure and function are limited to traditional imaging techniques and the mechanism for MI therapy is not clear. By observing the brain activation mode during MI and motor execution (ME) in healthy adults, this study aims to use near-infrared brain imaging technology to provide theoretical basis for the treatment of MI. METHODS: A total of 30 healthy adults recruited to the public from June 2021 to August 2021. The MI and ME of the right knee movement served as the task mode. Block design was repeated 5 times alternately in a 20 s task period and a 30 s resting period. The activation patterns of brain regions were compared between the 2 tasks, and the regression coefficient was calculated to reflect the activation intensity of each brain region by Nirspark and SPSS 23.0 softwares. RESULTS: Lane 2, 3, 4, 5, 7, 9, 19, 20, 21, 24, 25, 26, 27, 32, 33, and 34 were significantly activated during the ME task (P<0.05, corrected by FDR) and lane 2, 5, 9, 16, 27, 29, 33, 34, and 35 were significantly activated during the MI task (P<0.05, corrected by FDR). According to the channel brain region registration information, the brain region activation pattern was similar during both MI and ME tasks in healthy adults, including left primary motor cortex (LM1), left primary sensory cortex (LS1), prefrontal pole, Broca area, and right supramarginal gyrus. Both LM1 and left pre-motor cortex (LPMC) were activated during MI in healthy adults, whereas dorsolateral prefrontal cortex (DLPFC) and only LM1 of the motor region were activated during ME. Compared to MI, the activation intensity of left sensory and left motor cortex was significantly enhanced in ME, and that of left and right prefrontal cortex especially left and right pars triangularis Broca's area (P<0.001, corrected by FDR) were significantly enhanced. CONCLUSIONS: The rationality of MI therapy is proved by functional near-infrared spectroscopy. The involvement of DLPFC in motor decision-making may regulate the two-way feedback of premoter cortex-M1 during ME; and Broca area, closely related to the motor program understanding, participates in MI and ME.

Dock1 promotes the mesenchymal transition of glioma and is modulated by MiR-31.

AIMS: This research aimed to examine the relationship between Dock1 and miR-31 and to determine the effect of miR-31 on the mesenchymal transition and invasiveness of glioma. METHODS: Real-time PCR was used to measure the expression of miR-31 and other RNAs. The transfection was used to manipulate the expression levels of Dock1 and miR-31 in cancer cells. Western blot was used to detect the expression of Dock1 and other related proteins. Wound healing, Matrigel invasion and chemotaxis assays were performed to detect the invasion and migration of glioma cell lines. The actual binding site of miR-31 to the 3'-untranslated region of Dock1 was confirmed through luciferase assay and RNA immunoprecipitation. Methylation-specific PCR was performed to detect the methylation level of miR-31 in both glioma cell lines and tissues. RESULTS: Dock1 can promote the IL8-induced chemotaxis and mesenchymal transition of glioma cells through the NF-κB/Snail signalling pathway. The protein levels of Dock1 in glioma cell lines and clinical specimens were negatively correlated with miR-31 expression, and Dock1 was directly targeted by miR-31. Animal experiments showed that Dock1 downregulation and miR-31 overexpression reduced glioma cell invasion. Investigation of the underlying molecular mechanism revealed that miR-31 downregulation was attributable to the hypermethylation of the promoter region of miR-31 in glioma cells. CONCLUSION: Dock1 modulation by miR-31 plays an important function in glioma invasion both in vitro and in vivo. This study provides new insights into the invasion of glioma cells and might therefore contribute to the development of new antiglioma strategies.

Application of microfluidic devices in studies of thrombosis and hemostasis.

Due to the importance of fluid flow during thrombotic episodes, it is quite appropriate to study clotting and bleeding processes in devices that have well-defined fluid shear environments. Two common devices for applying these defined shear stresses include the cone-and-plate viscometer and parallel-plate flow chamber. While such tools have many salient features, they require large amounts of blood or other protein components. With growth in the area of microfluidics over the last two decades, it has become feasible to miniaturize such flow devices. Such miniaturization not only enables saving of precious samples but also increases the throughput of fluid shear devices, thus enabling the design of combinatorial experiments and making the technique more accessible to the larger scientific community. In addition to simple flows that are common in traditional flow apparatus, more complex geometries that mimic stenosed arteries and the human microvasculature can also be generated. The composition of the microfluidics cell substrate can also be varied for diverse basic science investigations, and clinical investigations that aim to assay either individual patient coagulopathy or response to anti-coagulation treatment. This review summarizes the current state of the art for such microfluidic devices and their applications in the field of thrombosis and hemostasis.

[Effect of high frequency electrotherapy on caspase-3 and ultra microstructure of hippocampus in rats following cerebral ischemia/reperfusion].

OBJECTIVE: To investigate the effect of high frequency electrotherapy (HFE) on rat hippocampus after cerebral ischemia/reperfusion (I/R).
 Methods: A rat model of cerebral I/R injury was established. The rats were randomly divided into a sham group, an I/R group and an HFE group. The HFE group received thearapy daily for different sessions for 1, 3, 7 d. Neuronal deficit score,neuron ultra microstructure in the hippocampus and caspase-3 protein expression were measured on 1 st, 3 th and 7th d.
 Results: Compared with the I/R group, the HFE group showed the decreased neurological deficit scores, with significant differences between the 2 groups (P<0.05). The injury in HFE group was reduced compared with that in the I/R group based on the electron microscope test, with significant difference. Caspase-3 protein in brain tissue in the HFE group also downregulated compared with that in the I/R group (P<0.05).
 Conclusion: High frequency electrotherapy can improve neural function, suppress caspase-3 expression and apoptosis in nerve cells and improve the ultra microstructure of neurons, displaying a protective effect on cerebral I/R injury in rats.

Explore the variation of MMP3, JNK, p38 MAPKs, and autophagy at the early stage of osteoarthritis.

Osteoarthritis is a chronic disease characterized by cartilage degeneration and chondrocyte apoptosis. Mitogen-activated protein kinase (MAPK) signaling pathway plays a key role in regulating OA process. Autophagy has an important effect on the OA process, and it is believed to be regulated by MAPKs. To reveal the mechanism and the effect of JNK and p38 MAPKs on matrix metalloproteinase 3 (MMP3) and autophagy in OA, the study established OA model in rabbits, used the measurement of the Osteoarthritis Research Society International scoring system to evaluate OA model, and conducted general observation, histological observation, and Western blotting of JNK, phosphorylate-JNK (P-JNK), p38, phosphorylate-p38 (P-p38), MMP3, and light-chain 3 (LC3)-II/LC3-I to explore the variation of JNK, p38 MAPKs, and autophagy at the early stage of OA. With OA progressing at the early stage, MMP3, P-p38, and P-JNK were gradually upregulated from the baseline to the peak in study groups when compared with the control group; JNK and p38 variated of turbulence without statistical difference; and LC3-II/LC3-I had a decreasing tendency from the 0- to 15-day group. This study identifies that compromised autophagy may be related to the OA progress and that JNK and p38 MAPKs have positive regulation on MMP3 and negative regulation on autophagy. It also implicates a new therapeutic strategy for OA and other degenerate diseases based on selective MAPK inhibitors, reduction of MMP3, and autophagy.

A New Approach of Short Wave Protection against Middle Cerebral Artery Occlusion/Reperfusion Injury via Attenuation of Golgi Apparatus Stress by Inhibition of Downregulation of Secretory Pathway Ca(2+)-ATPase Isoform 1 in Rats.

BACKGROUND: Short wave (SW), a pattern of electromagnetic therapy, achieves an oscillating electromagnetic field. It has been reported that it may have a potential effect on cerebral injury. The present study was designed to investigate the potential role and possible mechanism of SW in focal cerebral ischemia/reperfusion (I/R) injury in rats. Secretory pathway Ca(2+)/Mn(2+) ATPase isoform 1 is a major component of Golgi apparatus stress. It has been reported as representative of Golgi apparatus stress. METHODS: Up to 120 minutes of middle cerebral artery occlusion (MCAO) and reperfusion injury was induced in male Sprague-Dawley rats. Different sessions of SW daily were administered over head after reperfusion from day 1 to day 7. Functional recovery scores, survival rates, infarct volume analysis, electron microscope test, and western blotting studies were used to analyze the therapy. RESULTS: SW protected against neuronal death and apoptosis in cornu ammon 1 region of hippocampus by reducing neuronal deficit, infarct volume, and ultrastructure. SW partly inhibited upregulation of caspase3. In addition, the expression of secretory pathway Ca(2+)-ATPase isoform 1 (SPCA1) was upregulated by SW. CONCLUSIONS: Our data indicate that SW can be protected against focal cerebral I/R injury, and the influence on Golgi apparatus stress might provide us a new perspective in further study. To the authors' knowledge, this is the first report using SW to increase expression of SPCA1 indicating modulate Golgi apparatus stress in MCAO and reperfusion model.

[Reliability and validity of the Chinese Eating Assessment Tool (EAT-10) in evaluation of acute stroke patients with dysphagia].

OBJECTIVE: To study the reliability and validity of the Chinese Eating Assessment Tool (EAT-10) in evaluation of acute stroke patients with dysphagia.
 METHODS: The inpatients of stroke were assessed with Chinese EAT-10. As a golden standard for evaluation of dysphagia, videofluoroscopic swallow study (VFSS) test was used to judge the reliability and validity of EAT-10.
 RESULTS: A total of 130 qualified questionnaires were collected. The Cronbach's alpha coefficient for Chinese EAT-10 scale was 0.845. The total score of each item was related. The lowest or highest correlation coefficient for the item 2 or 3 was 0.271 or 0.772. The retest reliability was greater than 0.7, which met the requirements. According to the investigator consistency reliability test, the value collected from the investigator in the item 2 kept constant. The consistent correlation coefficient of the remaining nine items was more than 0.7. The consistency between each item and the mean score was high. The EAT-10 with the cut-off point at 1 was an optimal cut-off point. With the cut-off value of 1 (EAT-10 score ≥ 1), the sensitivity and specificity for EAT-10 was 77.9% and 66.1%, respectively. The positive predictive value (PPV) and negative value (NPV) was 71.6% and 73.2%, respectively, with 2.30 LHR+ and 0.33 LHR- for dysphagia. 
 CONCLUSION: The Chinese EAT-10 has a good reliability and validity in evaluation of the acute stroke patients with dysphagia. The sensitivity and negative value are the best with the cut-off value of 1 (EAT-10 score ≥ 1). It offers a good way to discriminate dysphagia, impaired efficacy, penetrations, and aspirations in acute stroke patients.

Muscle activation patterns and gait changes in unilateral knee osteoarthritis patients: a comparative study with healthy controls.

The objective of this study was to investigate the differences in muscle activation and kinematic parameters between patients with unilateral knee osteoarthritis (OA) and healthy individuals. Additionally, the study aimed to determine the correlation between muscle activation and kinematic parameters with knee OA symptoms. Participants with unilateral knee OA (n = 32) and healthy individuals (n = 32) completed the gait test. Electromyography (EMG) and motion capture were employed to collect muscle activation data and kinematic parameters. Spearman's correlation coefficient was used to analysis the correlation between BMI, symptomatic side EMG parameters, kinematic parameters, and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores. Furthermore, a multiple linear regression analysis of WOMAC pain was also conducted. The peak root mean square, integrated electromyography, and co-activation index (CCI) were increased bilaterally in the unilateral knee OA group compared to the healthy group. Furthermore, these values were higher on the symptomatic side than on the asymptomatic side. Compared with the healthy group, the knee OA group had lower gait speed, decreased stride length and cadence on bilateral sides, longer total stance time and double-stance time, and shorter single stance time and swing time. The maximum knee flexion angle of the swing phase on the symptomatic side of the knee OA group was smaller than that on the asymptomatic side and healthy group. Changes in EMG and gait parameters on the symptomatic side correlated with WOMAC scores. The main factors influencing WOMAC pain were the CCI values of the lateral femoral and biceps femoris muscles and gait speed. Muscle activation and kinematic parameters in the lower limbs of patients with unilateral knee OA were altered bilaterally during walking. These alterations on the symptomatic side were associated with knee OA-related pain. ChiCTR2200064958. Date of registration: 2022-10-24. Key Points • Unilateral symptomatic knee OA leads to bilateral alterations in muscle activation and gait parameters. • Symptomatic muscle activation and gait parameter changes in knee OA patients are associated with knee OA symptoms. • Correcting abnormal muscle activation conditions and gait training may reduce knee OA-related pain.

Spatiotemporal evolution of deformation and LSTM prediction model over the slope of the deep excavation section at the head of the South-North Water Transfer Middle Route Canal.

Slope deformation is one of the focal issues of concern during the normal operation and maintenance of the South-North Water Transfer Middle Route Project. To study the slope deformation evolution in the deep excavation section at the head of the canal, we applied 88 views of Sentinel-1A ascending image data from 2017 to 2019 and MT-InSAR(Multi-temporal InSAR) deformation monitoring technology to obtain long-time series deformation rates and cumulative deformation fields over the slope in the study area. Based on the analysis of the time-series monitoring data of the deformation field sample points, a LSTM (Long Short Term Memory Network) slope deformation predictive model was constructed to predict the slope deformation for the next 12 months at 12 sample points of the deep excavation slope. The impact of rainfall on slope deformation was investigated, and the reliability of the LSTM model was verified by using the measured data. The results show that the average annual deformation rate of the slope ranges from 10mm/a to 25mm/a, the maximum cumulative deformation is about 60 mm, and the slope of the excavated section is generally in an uplifted state. The rainfall-induced repeated uplift or subsidence of the canal slopes together with the peak deformation was closely related to the amount of rainfall during the wet season, and the longer the duration of the wet season, the more obvious the crest. Among the12 sample sites, the minimum and maximum deformation predicted using the LSTM model were 51.7 mm and 73.9 mm respectively, with the lowest correlation coefficient of 0.994 and the highest of 0.999. The maximum and minimum values of RMSE (Root Mean Square Error) were 4.4 mm and 3.6 mm respectively, indicating reliable prediction results. The results of the study can provide reference for the prevention and control of geological hazards in the South-North Water Transfer Canal.

Primary Cilia: A Cellular Regulator of Articular Cartilage Degeneration.

Osteoarthritis (OA) is the most common joint disease that can cause pain and disability in adults. The main pathological characteristic of OA is cartilage degeneration, which is caused by chondrocyte apoptosis, cartilage matrix degradation, and inflammatory factor destruction. The current treatment for patients with OA focuses on delaying its progression, such as oral anti-inflammatory analgesics or injection of sodium gluconate into the joint cavity. Primary cilia are an important structure involved in cellular signal transduction. Thus, they are very sensitive to mechanical and physicochemical stimuli. It is reported that the primary cilia may play an important role in the development of OA. Here, we review the correlation between the morphology (location, length, incidence, and orientation) of chondrocyte primary cilia and OA and summarize the relevant signaling pathways in chondrocytes that could regulate the OA process through primary cilia, including Hedgehog, Wnt, and inflammation-related signaling pathways. These data provide new ideas for OA treatment.

Exosomal miR-150 derived from BMSCs inhibits TNF-α-mediated osteoblast apoptosis in osteonecrosis of the femoral head by GREM1/NF-κB signaling.

Aim: The purpose of this study was to investigate the functions of exosomal miR-150 derived from bone marrow mesenchymal stem cells in osteonecrosis of the femoral head (ONFH). Materials & methods: Cell viability and apoptosis were detected using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry. Alizarin red staining was performed to detect calcium deposits. A rat model was established to assess the effects of exosomal miR-150 on ONFH in vivo. Results: Exosomes or exosomal miR-150 derived from bone marrow mesenchymal stem cells inhibited TNF-α-induced osteoblast apoptosis and promoted osteogenic differentiation and autophagy. Exosomal miR-150 suppressed apoptosis and induced autophagy in TNF-α-treated osteoblasts by regulating the GREM1/NF-κB axis. Exosomal miR-150 also improved the pathological features of ONFH in vivo. Conclusion: Exosomal miR-150 alleviates ONFH by mediating the GREM1/NF-κB axis. This study provides a potential therapeutic strategy for ONFH.

Osteonecrosis of the femoral head (ONFH) is an orthopedic disease that frequently occurs in young adults aged less than 50 years. At present, there is no widely accepted curative surgical procedure or drug therapy for this disease. Bone marrow mesenchymal stem cells (BMSCs) play a key role in the progression of ONFH. BMSC-derived exosomes refer to small membrane vesicles that can transfer proteins, miRNAs and mRNAs, which are closely related to the development of ONFH. This study showed that exosomal miRNA-150 derived from BMSCs inhibited TNF-α-induced osteoblast apoptosis and promoted osteogenic differentiation and autophagy by regulating the GREM1/NF-κB axis. In addition, exosomal miRNA-150 alleviated the symptoms of ONFH in rats.

MicroRNA-30a-5p inhibits the proliferation and invasion of gastric cancer cells by targeting insulin-like growth factor 1 receptor.

[This retracts the article DOI: 10.3892/etm.2017.4477.].

Face masks matter: Eurasian tree sparrows show reduced fear responses to people wearing face masks during the COVID-19 pandemic.

Continuous exposure to human activity has led to considerable behavioural changes in some wildlife populations. Animals are more likely to survive in a changing environment by adjusting their behaviour to repeatedly occurring but harmless stimulations. During the COVID-19 pandemic, starting in late 2019, face masks were recommended to the public to prevent the spread of pathogens. In this context, we compared the flight initiation distance (FID) of the Eurasian tree sparrow (Passer montanus), a commonly seen bird across China, in Yibin and Dazhou, Sichuan, in response to people with or without face masks. After continuous exposure to people wearing face masks for nearly six months, sparrows evidently became adapted to people wearing face masks, and correspondingly showed shorter FIDs in response to people wearing masks. To our knowledge, this is the first study showing that birds show reduced fear responses to people wearing face masks during the COVID-19 pandemic. Our results suggest a novel aspect of short-term adaptation of wildlife to human behaviour, and that the learning ability of sparrows allows them to adjust their behaviours to adapt to such subtle changes in the environment.

The neural correlates of Chinese children's spontaneous trait inferences: Behavioral and electrophysiological evidence.

The results of multiple behavioral investigations indicate that children can develop spontaneous trait inferences (STIs) starting from when they are 8-9 years of age. However, little is known about the neural correlates of STIs when they first develop. In the present study, we measured event-related potentials (ERPs) and event-related spectral perturbation (ERSP) using an N400 paradigm. Seventeen 9-year-old Chinese children were first instructed to remember facial photos and paired trait-implying behaviors. Subsequently, they performed a lexical decision task in which faces were used to prime the inferred traits or their antonyms. Compared to the inferred traits, the antonyms exhibited a stronger amplitude on both N400 (370-500 ms) and negative slow wave (NSW; 550-800 ms). However, only N400 showed a right hemispheric dominance of STIs. In addition, ERSP results revealed stronger lower alpha band (7-10 Hz) activity for antonyms than inferred traits over the right parieto-occipital area from 420 ms to 800 ms. These findings, coupled with the electroencephalography (EEG) source localization (standardized low-resolution electromagnetic tomography [sLORETA]), provide evidence for a "monitoring-control" two-stage neural network.

Prevalence and risk factors of disability and anxiety in a retrospective cohort of 432 survivors of Coronavirus Disease-2019 (Covid-19) from China.

OBJECTIVE: To estimate the prevalence of disability and anxiety in Covid-19 survivors at discharge from hospital and analyze relative risk by exposures. DESIGN: Multi-center retrospective cohort study. SETTING: Twenty-eight hospitals located in eight provinces of China. METHODS: A total of 432 survivors with laboratory-confirmed SARS CoV-2 infection participated in this study. At discharge, we assessed instrumental activities of daily living (IADL) with Lawton's IADL scale, dependence in activities of daily living (ADL) with the Barthel Index, and anxiety with Zung's self-reported anxiety scale. Exposures included comorbidity, smoking, setting (Hubei vs. others), disease severity, symptoms, and length of hospital stay. Other risk factors considered were age, gender, and ethnicity (Han vs. Tibetan). RESULTS: Prevalence of at least one IADL problem was 36.81% (95% CI: 32.39-41.46). ADL dependence was present in 16.44% (95% CI: 13.23-20.23) and 28.70% (95% CI: 24.63-33.15) were screened positive for clinical anxiety. Adjusted risk ratio (RR) of IADL limitations (RR 2.48, 95% CI: 1.80-3.40), ADL dependence (RR 2.07, 95% CI 1.15-3.76), and probable clinical anxiety (RR 2.53, 95% CI 1.69-3.79) were consistently elevated in survivors with severe Covid-19. Age was an additional independent risk factor for IADL limitations and ADL dependence; and setting (Hubei) for IADL limitations and anxiety. Tibetan ethnicity was a protective factor for anxiety but a risk factor for IADL limitations. CONCLUSION: A significant proportion of Covid-19 survivors had disability and anxiety at discharge from hospital. Health systems need to be prepared for an additional burden resulting from rehabilitation needs of Covid-19 survivors.

von Willebrand factor self-association is regulated by the shear-dependent unfolding of the A2 domain.

von Willebrand factor (VWF) self-association results in the homotypic binding of VWF upon exposure to fluid shear. The molecular mechanism of this process is not established. In this study, we demonstrate that the shear-dependent unfolding of the VWF A2 domain in the multimeric protein is a major regulator of protein self-association. This mechanism controls self-association on the platelet glycoprotein Ibα receptor, on collagen substrates, and during thrombus growth ex vivo. In support of this, A2-domain mutations that prevent domain unfolding due to disulfide bridging of N- and C-terminal residues ("Lock-VWF") reduce self-association and platelet activation under various experimental conditions. In contrast, reducing assay calcium concentrations, and 2 mutations that destabilize VWF-A2 conformation by preventing coordination with calcium (D1498A and R1597W VWD type 2A mutation), enhance self-association. Studies using a panel of recombinant proteins that lack the A1 domain ("ΔA1 proteins") suggest that besides pure homotypic A2 interactions, VWF-A2 may also engage other protein domains to control self-association. Addition of purified high-density lipoprotein and apolipoprotein-A1 partially blocked VWF self-association. Overall, similar conditions facilitate VWF self-association and ADAMTS13-mediated proteolysis, with low calcium and A2 disease mutations enhancing both processes, and locking-A2 blocking them simultaneously. Thus, VWF appears to have evolved 2 balancing molecular functions in a single A2 functional domain to dynamically regulate protein size in circulation: ADAMTS13-mediated proteolysis and VWF self-association. Modulating self-association rates by targeting VWF-A2 may provide novel methods to regulate the rates of thrombosis and hemostasis.

Genetic ablation of TAZ induces HepG2 liver cancer cell apoptosis through activating the CaMKII/MIEF1 signaling pathway.

BACKGROUND AND OBJECTIVE: Transcriptional coactivator with PDZ-binding motif (TAZ) has been found to be associated with tumor progression. Mitochondrial homeostasis regulates cancer cell viability and metastasis. However, the roles of TAZ and mitochondrial homeostasis in liver cancer viability have not been explored. The aim of our study was to investigate the influence of TAZ on HepG2 liver cancer cell apoptosis. MATERIALS AND METHODS: HepG2 liver cancer cell was used in the present study, and shRNA against TAZ was transfected into HepG2 cell to knockdown TAZ expression. Mitochondrial function was analyzed using Western blotting, immunofluorescence assay, and flow cytometry. Pathway blocker was used to confirm the role of CaMKII pathway in TAZ-mediated cancer cell death. RESULTS: Our results indicated that TAZ deletion induced death in HepG2 cell via apoptosis. Biological analysis demonstrated that mitochondrial stress, including mitochondrial bioenergetics disorder, mitochondrial oxidative stress, and mitochondrial apoptosis, were activated by TAZ deletion. Furthermore, we found that TAZ affected mitochondrial stress by triggering mitochondrial elongation factor 1 (MIEF1)-related mitochondrial dysfunction. The loss of MIEF1 sustained mitochondrial function and promoted cancer cell survival. Molecular investigation illustrated that TAZ regulated MIEF1 expression via the CaMKII signaling pathway. Blockade of the CaMKII pathway prevented TAZ-mediated MIEF1 upregulation and improved cancer cell survival. CONCLUSION: Taken together, our results highlight the key role of TAZ as a master regulator of HepG2 liver cancer cell viability via the modulation of MIEF1-related mitochondrial stress and the CaMKII signaling pathway. These findings define TAZ and MIEF1-related mitochondrial dysfunction as tumor suppressors that act by promoting cancer apoptosis via the CaMKII signaling pathway, with potential implications for new approaches to liver cancer therapy.

Microclot array elastometry for integrated measurement of thrombus formation and clot biomechanics under fluid shear.

Blood clotting at the vascular injury site is a complex process that involves platelet adhesion and clot stiffening/contraction in the milieu of fluid flow. An integrated understanding of the hemodynamics and tissue mechanics regulating this process is currently lacking due to the absence of an experimental system that can simultaneously model clot formation and measure clot mechanics under shear flow. Here we develop a microfluidic-integrated microclot-array-elastometry system (clotMAT) that recapitulates dynamic changes in clot mechanics under physiological shear. Treatments with procoagulants and platelet antagonists and studies with diseased patient plasma demonstrate the ability of the system to assay clot biomechanics associated with common antiplatelet treatments and bleeding disorders. The changes of clot mechanics under biochemical treatments and shear flow demonstrate independent yet equally strong effects of these two stimulants on clot stiffening. This microtissue force sensing system may have future research and diagnostic potential for various bleeding disorders.