SUMOylation of Rho-associated protein kinase 2 induces goblet cell metaplasia in allergic airways.

Allergic asthma is characterized by goblet cell metaplasia and subsequent mucus hypersecretion that contribute to the morbidity and mortality of this disease. Here, we explore the potential role and underlying mechanism of protein SUMOylation-mediated goblet cell metaplasia. The components of SUMOylaion machinery are specifically expressed in healthy human bronchial epithelia and robustly upregulated in bronchial epithelia of patients or mouse models with allergic asthma. Intratracheal suppression of SUMOylation by 2-D08 robustly attenuates not only allergen-induced airway inflammation, goblet cell metaplasia, and hyperreactivity, but IL-13-induced goblet cell metaplasia. Phosphoproteomics and biochemical analyses reveal SUMOylation on K1007 activates ROCK2, a master regulator of goblet cell metaplasia, by facilitating its binding to and activation by RhoA, and an E3 ligase PIAS1 is responsible for SUMOylation on K1007. As a result, knockdown of PIAS1 in bronchial epithelia inactivates ROCK2 to attenuate IL-13-induced goblet cell metaplasia, and bronchial epithelial knock-in of ROCK2(K1007R) consistently inactivates ROCK2 to alleviate not only allergen-induced airway inflammation, goblet cell metaplasia, and hyperreactivity, but IL-13-induced goblet cell metaplasia. Together, SUMOylation-mediated ROCK2 activation is an integral component of Rho/ROCK signaling in regulating the pathological conditions of asthma and thus SUMOylation is an additional target for the therapeutic intervention of this disease.

Orthogonal Design and Microstructure Mechanism Analysis of Novel Bentonite Polymer Slurry in Pipe Jacking.

The selection of an appropriate slurry ingredient and its percentage ratio is a vital and necessary task for engineers in slurry pipe jacking operations. However, traditional bentonite grouting materials are difficult to degrade because of their single and non-biodegradable composition. Nowadays crosslinked polymers have been widely considered due to their excellent performance and application in engineering practices, which enlighten novel polymer slurry in pipe jacking. This study innovatively proposed using boric acid crosslinked polymers added into polyacrylamide bentonite slurry, which not only solves the shortcomings of traditional grouting materials but also meets the general working performance requirements. The new slurry's funnel viscosity, filter loss, water dissociation ratio and dynamic shear were tested according to an orthogonal experiment. Single factor range analysis was conducted to identify the optimal mix proportion based on an orthogonal design, and the formation behavior of mineral crystals and microstructure characteristics were evaluated by X-ray diffraction and scanning electron microscopy respectively. According to the results, guar gum and borax form a dense boric acid crosslinked polymer through cross-linking reaction. The internal structure grew tighter and more continuous as the crosslinked polymer concentration grew. It improved the anti-permeability plugging action and viscosity of slurries by 36.1~94.3%. The optimal proportions of sodium bentonite, guar gum, polyacrylamide, borax, and water were 10%, 0.2%, 0.25%, 0.1%, and 89.45% respectively. All these works indicated that the improvement of slurry composition by using boric acid crosslinked polymers was feasible.

Promising therapeutic aspects in human genetic imprinting disorders.

Genomic imprinting is an epigenetic phenomenon of monoallelic gene expression pattern depending on parental origin. In humans, congenital imprinting disruptions resulting from genetic or epigenetic mechanisms can cause a group of diseases known as genetic imprinting disorders (IDs). Genetic IDs involve several distinct syndromes sharing homologies in terms of genetic etiologies and phenotypic features. However, the molecular pathogenesis of genetic IDs is complex and remains largely uncharacterized, resulting in a lack of effective therapeutic approaches for patients. In this review, we begin with an overview of the genomic and epigenomic molecular basis of human genetic IDs. Notably, we address ethical aspects as a priority of employing emerging techniques for therapeutic applications in human IDs. With a particular focus, we delineate the current field of emerging therapeutics for genetic IDs. We briefly summarize novel symptomatic drugs and highlight the key milestones of new techniques and therapeutic programs as they stand today which can offer highly promising disease-modifying interventions for genetic IDs accompanied by various challenges.

Dysregulated adipose tissue expansion and impaired adipogenesis in Prader-Willi syndrome children before obesity-onset.

OBJECTIVE: Prader-Willi syndrome (PWS) is a rare genetic imprinting disorder resulting from the expression loss of genes on the paternally inherited chromosome 15q11-13. Early-onset life-thriving obesity and hyperphagia represent the clinical hallmarks of PWS. The noncoding RNA gene SNORD116 within the minimal PWS genetic lesion plays a critical role in the pathogenesis of the syndrome. Despite advancements in understanding the genetic basis for PWS, the pathophysiology of obesity development in PWS remains largely uncharacterized. Here, we aimed to investigate the signatures of adipose tissue development and expansion pathways and associated adipose biology in PWS children without obesity-onset at an early stage, mainly from the perspective of the adipogenesis process, and further elucidate the underlying molecular mechanisms. METHODS: We collected inguinal (subcutaneous) white adipose tissues (ingWATs) from phase 1 PWS and healthy children with normal weight aged from 6 M to 2 Y. Adipose morphology and histological characteristics were assessed. Primary adipose stromal vascular fractions (SVFs) were isolated, cultured in vitro, and used to determine the capacity and function of white and beige adipogenic differentiation. High-throughput RNA-sequencing (RNA-seq) was performed in adipose-derived mesenchymal stem cells (AdMSCs) to analyze transcriptome signatures in PWS subjects. Transient repression of SNORD116 was conducted to evaluate its functional relevance in adipogenesis. The changes in alternative pre-mRNA splicing were investigated in PWS and SNORD116 deficient cells. RESULTS: In phase 1 PWS children, impaired white adipose tissue (WAT) development and unusual fat expansion occurred long before obesity onset, which was characterized by the massive enlargement of adipocytes accompanied by increased apoptosis. White and beige adipogenesis programs were impaired and differentiated adipocyte functions were disturbed in PWS-derived SVFs, despite increased proliferation capacity, which were consistent with the results of RNA-seq analysis of PWS AdMSCs. We also experimentally validated disrupted beige adipogenesis in adipocytes with transient SNORD116 downregulation. The transcript and protein levels of PPARγ, the adipogenesis master regulator, were significantly lower in PWS than in control AdMSCs as well as in SNORD116 deficient AdMSCs/adipocytes than in scramble (Scr) cells, resulting in the inhibited adipogenic program. Additionally, through RNA-seq, we observed aberrant transcriptome-wide alterations in alternative RNA splicing patterns in PWS cells mediated by SNORD116 loss and specifically identified a changed PRDM16 gene splicing profile in vitro. CONCLUSIONS: Imbalance in the WAT expansion pathway and developmental disruption are primary defects in PWS displaying aberrant adipocyte hypertrophy and impaired adipogenesis process, in which SNORD116 deficiency plays a part. Our findings suggest that dysregulated adiposity specificity existing at an early phase is a potential pathological mechanism exacerbating hyperphagic obesity onset in PWS. This mechanistic evidence on adipose biology in young PWS patients expands knowledge regarding the pathogenesis of PWS obesity and may aid in developing a new therapeutic strategy targeting disturbed adipogenesis and driving AT plasticity to combat abnormal adiposity and associated metabolic disorders for PWS patients.

A self-efficacy-enhancing physical activity intervention in women with high-risk factors for gestational diabetes mellitus: study protocol for a randomized clinical trial.

BACKGROUND: Gestational diabetes mellitus (GDM) is one of the most common medical disorders in pregnancy. Evidence has demonstrated that moderate-intensity physical activity may reduce the risk of gestational diabetes. However, women at risk of GDM spend most of their time performing sedentary behaviors. Although researchers identified self-efficacy as a mediator to overcome physical activity barriers, exercise intervention during pregnancy based on self-efficacy theory has not been discussed so far. Furthermore, there is conflicting evidence regarding the effects of a physical exercise intervention on the incidence of GDM and other maternal or neonatal outcomes in women at higher risk for GDM. METHODS/DESIGN: A single-center, parallel, randomized controlled trial will be conducted in a maternal-child health care center. A total of 244 pregnant women at high risk for GDM will be randomized into a study group receiving a self-efficacy-enhancing physical activity intervention or a control group receiving the usual care. The intervention will consist of four group sessions and everyday reminders by WeChat (Tencent, Shenzhen, China). The program will begin at approximately 13-14+6 gestational weeks and end at 36+6 gestational weeks. The primary outcomes will include the incidence of GDM, blood sugar values, and physical activity. The secondary outcomes will include physical activity self-efficacy, gestational weight gain, maternal outcomes, and neonatal outcomes. DISCUSSION: The findings of this research will contribute toward understanding the effects of a self-efficacy theory-oriented physical activity program on the incidence of GDM, blood sugar values, physical activity level, gestational weight gain, physical activity self-efficacy, maternal outcomes, and neonatal outcomes. TRIAL REGISTRATION: Chinese Clinical Trial Registry (CHiCTR) ChiCTR2200056355 . Registered on February 4, 2022.

Experimental investigations into the performance of die-sinking mixed-gas atomization discharge ablation process on titanium alloy.

As a variant of highly efficient electrical discharge machining (EDM), the die-sinking mixed-gas atomization discharge ablation process (DMA-DAP) uses an atomized dielectric formed by a mixed gas, which mainly composed of oxygen and supplemented by nitrogen, and water medium as the discharge medium. In this technology, the oxygen in the medium is used for exothermic oxidation, and the vaporization and explosion of the water generates a chip removal force for highly efficient erosion. The present work uses single-factor tests to compare the characteristics of processing the difficult-to-machine material titanium-alloy special-shaped cavities using either DMA-DAP or EDM. The current, pulse width, pulse interval, and dielectric pressure are selected as the single-factor processing parameters, and how they influence the material removal rate (MRR), electrode relative wear rate (ERWR) and the surface morphology of the processed square cavities is analyzed. The results show that with DMA-DAP, the MRR is more than 12 times that of EDM, the ERWR is reduced by more than 98%, and the surface morphology is relatively good. Finally, taking an aero-engine radial diffuser as the profiling object, DMA-DAP realizes a profiling sample in the form of a variable-cross-section cavity that EDM cannot process, and the efficient die-sinking processing ability of DMA-DAP is verified.

CircZNF652 promotes the goblet cell metaplasia by targeting the miR-452-5p/JAK2 signaling pathway in allergic airway epithelia.

BACKGROUND: Circular RNAs (circRNAs) play potentially important roles in various human diseases; however, their roles in the goblet cell metaplasia of asthma remain unknown. OBJECTIVE: We sought to investigate the potential role and underlying mechanism of circZNF652 in the regulation of allergic airway epithelial remodeling. METHODS: The differential expression profiles of circRNAs were analyzed by transcriptome microarray, and the effects and mechanisms underlying circZNF652-mediated goblet cell metaplasia were investigated by quantitative real-time PCR, RNA fluorescence in situ hybridization, Western blot, RNA pull-down, and RNA immunoprecipitation analyses. The roles of circZNF652 and miR-452-5p in allergic airway epithelial remodeling were explored in both the mouse model with allergic airway inflammation and children with asthma. RESULTS: One hundred sixty circRNAs were differentially expressed in bronchoalveolar lavage fluid of children with asthma versus children with foreign body aspiration, and 52 and 108 of them were significantly upregulated and downregulated, respectively. Among them, circZNF652 was predominantly expressed and robustly upregulated in airway epithelia of both the children with asthma and the mouse model with allergic airway inflammation. circZNF652 promoted the goblet cell metaplasia by functioning as a sponge of miR-452-5p, which released the Janus kinase 2 (JAK2) expression and subsequently activated JAK2/signal transducer and activator of transcription 6 (STAT6) signaling in the allergic airway epithelia. In addition, epithelial splicing regulatory protein 1, a splicing factor, accelerated the biogenesis of circZNF652 by binding to its flanking intron to promote the goblet cell metaplasia in allergic airway epithelial remodeling. CONCLUSIONS: Upregulation of circZNF652 expression in allergic bronchial epithelia contributed to the goblet cell metaplasia by activating the miR-452-5p/JAK2/STAT6 signaling pathway; thus, blockage of circZNF652 or agonism of miR-452-5p provided an alternative approach for the therapeutic intervention of epithelial remodeling in allergic airway inflammation.

Association between short-term heart rate variability and blood coagulation in patients with breast cancer.

The purpose of this study was to investigate the relationship between heart rate variability (HRV), a non-invasive tool for evaluating autonomic function, and routine coagulation indices (RCIs) in patients with breast cancer (BC). Forty-six BC patients were enrolled in this study. Blood biochemistry tests were performed to extract RCIs, including prothrombin time (PT), activated partial thromboplastin time (APTT), and thrombin time (TT). Five-minute electrocardiograms were collected for analysis of HRV parameters (SDNN, RMSSD, LF, HF, LF n.u., HF n.u., LF/HF). Multiple linear regression models examined the relationship of HRV parameters with RCIs. RMSSD, LF n.u., HF n.u., LF/HF were significantly associated with PT. Specifically, the value of PT increased by 0.192 ± 0.091 or 0.231 ± 0.088 s, respectively for each 1 standard deviation (SD) increase in RMSSD or HF n.u.; it increased by 0.230 ± 0.088 or 0.215 ± 0.088 s, respectively for each 1 - SD decrease in LF n.u. or ln (LF/HF) (all P < 0.05). RMSSD was significantly associated with APTT, i.e., the value of APTT increased by 1.032 ± 0.470 s for each 1 - SD increase in RMSSD (P < 0.05). HRV parameters were associated with RCIs in patients with BC. These observations suggest that the autonomic nervous system and coagulation indices in BC patients are linked, potentially explaining the reason that they are both associated with the prognosis.

The effects of temperature shock on the treatment of high-concentration organic wastewater by an Fe0/GO-anaerobic system.

In order to make up for the defects of traditional anaerobic fermentation systems, such as low energy utilization rates and the slow growth and reproduction of microorganisms, an Fe0/GO (zero-valent iron/graphene oxide) anaerobic biological treatment system was used as a treatment process in this paper, and the impact of temperature shock on the system during the treatment of high-concentration organic wastewater was studied. The experimental results showed that temperature shock reduced the CODCr removal rate and gas production level in each system, but the Fe0/GO group maintained a higher level and had the highest CODCr degradation rate after shocking. After temperature shock, the acetic acid content in each system was higher (above 90%), and the volatile fatty acid (VFA) content in the Fe0/GO group was the lowest. The mixed liquor suspended solids (MLSS) in all systems decreased after impact; the decrease was less in the Fe0/GO group and the increase was largest after temperature recovery. After shocking, the extracellular polymer substance (EPS) protein (PN) and polysaccharide (PS) levels in each system were both low. After temperature recovery, the PN/PS ratio of the Fe0/GO group was the highest, showing a strong impact resistance to temperature.

[Analysis of prenatal phenotype and pathogenetic variant in a fetus with Papillorenal syndrome].

OBJECTIVE: To determine the carrier rate of deafness-related genetic variants among 53 873 newborns from Zhengzhou. METHODS: Heel blood samples of the newborns were collected with informed consent from the parents, and 15 loci of 4 genes related to congenital deafness were detected by microarray. RESULTS: In total 2770 newborns were found to carry deafness-related variants, with a carrier rate of 5.142%. 1325 newborns (2.459%) were found to carry heterozygous variants of the GJB2 gene, 1071 (1.988%) were found with SLC26A4 gene variants, 205 were found with GJB3 gene variants (0.381%), and 120 were found with 12S rRNA variants (0.223%). Five newborns have carried homozygous GJB2 variants, two have carried homozygous SLC26A4 variants, five have carried compound heterozygous GJB2 variants, and four have carried compound heterozygous SLC26A4 variants. 33 neonates have carried heterozygous variants of two genes at the same time. CONCLUSION: The carrier rate of deafness-related variants in Zhengzhou, in a declining order, is for GJB2, SLC26A4, GJB3 and 12S rRNA. The common variants included GJB2 235delC and SLC26A4 IVS7-2A>G, which are similar to other regions in China. To carry out genetic screening of neonatal deafness can help to identify congenital, delayed and drug-induced deafness, and initiate treatment and follow-up as early as possible.

Acupuncture plus rehabilitation for post-stroke depression: A protocol for systematic review and meta-analysis.

BACKGROUND: Post-stroke depression (PSD) is a common stroke complication that is characterized by hopelessness, anxiety, disordered sleep, and lowered responsiveness. Rehabilitation and acupuncture treatments are often combined to treat PSD; however, there has been no meta-analysis on their synergistic effect. Therefore, we aim to perform a systematic review and meta-analysis to estimate the effectiveness of acupuncture and rehabilitation in PSD treatment. METHODS: We will search the following electronic databases: PubMed, the Cochrane Library, EMBASE, the China National Knowledge Infrastructure, the Chinese Biomedical Literature Database, China Science and Technology Journal Database, and Wan Fang databases. We will include studies published between the database initiation and May 2020. Two reviewers will separately conduct study selection, data extraction, and risk of bias assessment. Disputes will be settled by consulting a third reviewer. Review Manager Software 5.3 will be employed for this meta-analysis. RESULTS: This systematic review will assess whether acupuncture combined with rehabilitation treatment is more effective than rehabilitation alone in the management of PSD. CONCLUSION: This systematic review will provide evidence regarding the synergistic effect of acupuncture and rehabilitation treatment for PSD.

IL-4/IL-13 upregulates Sonic hedgehog expression to induce allergic airway epithelial remodeling.

We have previously demonstrated that upregulation of Sonic hedgehog (SHH) expression in allergic airway epithelia essentially contributes to the goblet cell metaplasia and mucous hypersecretion. However, the mechanism underlying the upregulation of SHH expression remains completely unknown. In cultured human airway epithelial cells, IL-4/IL-13 but not IL-5 robustly induces the mRNA and protein expression of SHH and in turn activates SHH signaling by promoting the JAK/STAT6-controlling transcription of SHH gene. Moreover, intratracheal instillation of IL-4 and/or IL-13 robustly activates STAT6 and concomitantly upregulates SHH expression in mouse airway epithelia, whereas, in Club cell 10-kDa protein (CC10)-positive airway epithelial cells of children with asthma, activated STAT6 closely correlates with the increased expression of SHH and high activity of SHH signaling. Finally, intratracheal inhibition of STAT6 by AS-1517499 significantly diminished the allergen-induced upregulation of SHH expression, goblet cell phenotypes, and airway hyperresponsiveness, in an ovalbumin- or house dust mite-induced mouse model with allergic airway inflammation,. Together, upregulation of SHH expression by IL-4/IL-13-induced JAK/STAT6 signaling contributes to allergic airway epithelial remodeling, and this study thus provides insight into how morphogen signaling is coordinated with Th2 cytokine pathways to regulate tissue remodeling in chronic airway diseases.

Sonic hedgehog (Shh) and CC chemokine ligand 2 signaling pathways in asthma.

Asthma is a chronic inflammatory disease of the airways in which many cells are involved, including mast cells, eosinophils, T lymphocytes, and so on. During the process, many chemokines and mediators are released to engage in recruiting and activating eosinophils and other inflammatory cells. Also, some signaling pathways are involved in the pathobiology of asthma. Sonic hedgehog (Shh) is one of the members of hedgehog gene families. Shh signaling plays a critical role in the embryonic development, including the lung. Previous findings from our team reveal that Shh is involved in the asthma pathogenesis. Recombinant Shh could induce the CC chemokine ligand 2 (CCL2) overexpressing and Smo inhibitor GDC-O449 could inhibit CCL2 expression in airway epithelial cells, monocytes, or macrophages. Hence, we reviewed the effects of Shh and CCL2 signaling pathways, and the interaction between signaling pathways in asthma.

Inhibition of p21-activated kinase 1 attenuates the cardinal features of asthma through suppressing the lymph node homing of dendritic cells.

Dendritic cell (DC) trafficking from lung to the draining mediastinal lymph nodes (MLNs) is a key step for initiation of T cell responses in allergic asthma. In the present study, we investigate the role of DC-mediated airway inflammation after inhibition of p21-activated kinase-1 (PAK1), an effector of Rac and Cdc42 small GTPases, in the allergen-induced mouse models of asthma. Systemic administration of PAK1 specific inhibitor IPA-3 significantly attenuates not only the airway inflammation but also the airway hyperresponsiveness in a mouse model of ovalbumin-induced asthma. Specifically, intratracheal administration of low dosage of IPA-3 consistently decreases not only the airway inflammation but also the DC trafficking from lung to the MLNs. Importantly, intratracheal instillation of IPA-3-treated and ovalbumin-pulsed DCs behaves largely the same as that of either Rac inhibitor-treated and ovalbumin-pulsed DCs or Cdc42 inhibitor-treated and ovalbumin-pulsed DCs in attenuation of the airway inflammation in ovalbumin-challenged mice. Mechanistically, PAK1 is not involved in the maturation, apoptosis, antigen uptake, and T cell activation of cultured DCs, but PAK1 dose lie on the downstream of Rac and Cdc42 to regulate the DC migration toward the chemokine C-C motif chemokine ligand 19. Taken together, this study demonstrates that inhibition of PAK1 attenuates the cardinal features of asthma through suppressing the DC trafficking from lung to the MLN, and that interfere with DC trafficking by a PAK1 inhibitor thus holds great promise for the therapeutic intervention of allergic diseases.

Relationship Between Immune Parameters and Non-alcoholic Fatty Liver Disease in Obese Children.

OBJECTIVE: To investigate the relationship between immune parameters and non-alcoholic fatty liver disease (NAFLD) in obese children. DESIGN: Cross-sectional study. SETTING: Hospital-based study in Zhejiang Province, China between July to September 2015. PARTICIPANTS: A total of 117 obese children and 209 healthy non-obese children were studied as the obese and control groups. Depending on the severity of NAFLD, the obese group was divided into subgroups 1 (without NAFLD), 2 (with simple fatty liver) and 3 (with steatohepatitis). OUTCOME MEASURES: Glucose metabolism, lipid metabolism and immune parameters. RESULTS: In the obese group, body mass index (BMI), waist-and hip-circumferences, fasting insulin, Homeostasis model of assessment for insulin resistance (HOMA-IR), triglyceride, total cholesterol, low density lipoprotein cholesterol (LDL-C), apolipoprotein (Apo)B/ApoA1, alanine aminotransferase, uric acid, white blood cells, neutrophils percentage, platelet and interleukin (IL)-6 were significantly higher than those in the controls (P<0.05), while lower high density lipoprotein cholesterol and lymphocyte percentage were noted (P<0.05). IL-10 in the subgroup 3 was higher than those in the control group, subgroup 1 and 2 (P<0.05). Logistic regression analysis showed that BMI, LDL-C, HOMA-IR and IL-10 were independent factors of NAFLD (P<0.05). CONCLUSION: These results support a low-grade chronic inflammation in obese children. Moreover, obesity, dyslipidaemia and IR are risk factors while IL-10 may be a protective factor for NAFLD.

[Perioperative treatment for senile lumbar disease accompanied by internal disease].

OBJECTIVE: To investigate the surgical method and perioperative treatment for senile lumbar disease accompanied by internal disease. METHODS: From June 2000 to December 2003, the complete neurological and physical examinations were performed on the patients before operation, as treatment of internal diseases could improve the patients' conditions. Lumbar operations were performed on 125 patients, among whom 23 had simple lumbar disc herniation, 13 had lumbar spine stenosis, 81 had lumbar disc herniation with lumbar spine stenosis, and 8 had spondylolisthesis. The JOA score was 11.6 +/- 2.5. There were 3 patients undergoing fenestration + discectomy, 16 undergoing semi-laminectomy + discectomy, 82 undergoing total-laminectomy + discectomy, 5 undergoing total-laminectomy+discectomy+pedicle fixation, 11 undergoing lamina decompression+nerve-root pathway decompression, and 8 undergoing pedicle screw fixation+bone graft and fusion in spondylolisthesis. RESULTS: With an effective medical treatment, the internal disease produced little effect on the operation. Improved functions and bone fusions were observed after operation. According to the JOA standards, the average alleviation rate was 87.9%. CONCLUSION: Early neurological examination and proper treatment of internal diseases are the keys to the successful operation on the senile patients with lumbar disease. Radiological data are important in avoidance of mistaken diagnosis.