Objective To study the effect of transcutaneous electrical acupoint stimulation (TEAS) on emergence agitation in children undergoing cochlear implantation.Methods Sixty patients (age range,1-4 years) scheduled to undergo pediatric cochlear implantation were enrolled and randomized into the TEAS group (group T,n =30) that received continuous TEAS and the control group (group C,n =30) that did not receive TEAS.Anesthesia time,operation time,change in blood flow dynamics during the perioperative period,extubation time,and PACU stay time were recorded.The incidence of emergence agitation and postoperative nausea and vomiting (PONV) was monitored.Results No significant intergroup differences were observed in age,sex,weight,operation time,and PACU stay time (P ＞ 0.05).The MAP and HR in group T were lower than those in group C at different time points such as 30 min after the start of surgery,the end of the surgery,the extubation and 5 min after extubation,and the difference was statistically significant (P ＜ 0.05).The FPS-R,FLACC,and PAED scores in group T were lower than those in group C at different time points within 30 min after exmbation (including the time of extubation),and the difference was statistically significant (P ＜ 0.05).Extubation time,anesthesia time,and incidence of PONV were lower in group T than in group C,and the difference was statistically significant (P ＜ 0.05).Conclusion TEAS combined with general anesthesia significantly reduces the incidence of emergence agitation in children undergoing surgery for congenital hearing loss;it also helps control hypotension,reheve dramatic hemodynamic changes during the waking period,and reduce the incidence of PONV.

OBJECTIVETo investigate the mechanisms of lysophosphatidic acid (LPA) in stimulating invasion and metastatic colonization of ovarian cancer cells.

METHODSThe metastatic ability in vivo of ovarian cancer SK-OV3, HEY, OVCAR3, and IGROV1 cells was determined in tumor-bearing nude mouse models. Matrigel assay was used to detect the changes of response in vitro of ovarian cancer cells to LPA after Rac(-) or Rac(+) adenovirus treatment. LPA-induced Rho GTPase activation was detected by GST-fusion protein binding assay.

RESULTSThe peritoneal metastatic colonization assay showed overt metastatic colonization in mice receiving SK-OV3 and HEY cell inoculation, indicating that they are invasive cells. Metastatic colonization was not detected in animals receiving OVCAR3 and IGROV1 cells, indicating that these cells are non-invasive cells. In the matrigel invasion assay, exposure to LPA led to a notably greater migratory response in metastatic SK-OV3 and HEY cells (Optical density: SK-OV3 cells: 0.594±0.023 vs. 1.697±0.049, P<0.01; HEY cells: 0.804±0.070 vs. 1.851±0.095, P<0.01). But LPA did little in the non-metastatic OVCAR3 and IGROV1 cells (Optical density A: OVCAR3 cells: 0.336±0.017 vs. 0.374±0.007, P>0.05; IGROV1 cells: 0.491±0.036 vs. 0.479±0.061, P>0.05). LPA migratory responses of ovarian cancer cells were closely related to their metastatic colonization capabilities (r = 0.983, P<0.05). Rac(-) blocked the LPA response of invasive SK-OV3 and HEY cells (LPA-induced fold increase of cell migration: SK-OV3 cells: 2.988±0.095 vs. 0.997±0.100,P=0.01; HEY cells: 2.404±0.059 vs. 0.901±0.072, P=0.01). But Rac(+) confered the non-invasive cells with LPA response and invasion capability (LPA-induced fold increase of cell migration: OVCAR3 cells: 1.072±0.080 vs. 1.898±0.078, P<0.01; IGROV1 cells: 1.002±0.044 vs. 2.141±0.057, P<0.05). Among Rho GTPases, only Rac activation was different between ovarian cancer cell lines with different metastatic capability after LPA stimulation: Cdc42 could not be activated in both the invasive and non-invasive cell lines. RhoA could be activated in both the invasive and non-invasive cell lines. Rac could be activated by LPA in the invasive ovarian cancer cell lines. However, Rac could not be activated in the non-invasive cell lines.

CONCLUSIONLysophosphatidic acid stimulates invasion and metastasis of ovarian cancer cells through Rac activation.

Animals ; Cell Movement ; Female ; Humans ; Lysophospholipids ; metabolism ; Mice ; Ovarian Neoplasms ; metabolism ; Tumor Cells, Cultured ; rho GTP-Binding Proteins ; rhoA GTP-Binding Protein

Objective:To investigate clinical efficacy of narrow-band ultraviolet B (NB-UVB) irradiation around vitiliginous lesions in the treatment of refractory vitiligo.Methods:A total of 126 patients with refractory vitiligo were retrospectively collected from Department of Dermatology, the First Affiliated Hospital of Nanjing Medical University from June 2019 to November 2020. The patients were treated with NB-UVB irradiation around vitiliginous lesions after partial covering (perilesional irradiation group) , or conventional NB-UVB irradiation (conventional irradiation group) , twice a week for 3 consecutive months. After the treatment, the efficacy was evaluated. By using the propensity score method, the lesions in the 2 groups were matched at a ratio of 1∶1. Univariate and multivariate logistic regression analyses and stratified analysis were used to analyze the clinical efficacy of NB-UVB irradiation around vitiliginous lesions in the treatment of refractory vitiligo.Results:Totally, there were 420 skin lesions in the perilesional irradiation group and 257 in the conventional irradiation group, and 190 lesions were enrolled into each group by propensity-score matching. Before and after the matching, the response rates were both significantly higher in the perilesional irradiation group (71.9%, 67.9%, respectively) than in the conventional irradiation group (31.9%, 30.0%, respectively, both P < 0.05) . After the propensity-score matching, both univariate and multivariate logistic regression analyses showed significant differences in the efficacy between the perilesional irradiation group and conventional irradiation group ( OR = 4.9, 95% CI: 3.2, 7.6, P < 0.001; OR = 12.0, 95% CI: 6.5, 22.3, P < 0.001, respectively) . Vitiliginous lesions were classified according to hair types and irradiation methods: before the matching, there were 187 vitiliginous lesions with white hairs treated with the conventional irradiation and 246 treated with the perilesional irradiation, and there were 70 vitiliginous lesions with black hairs treated with the conventional irradiation and 174 treated with the perilesional irradiation; after the matching, 140 vitiliginous lesions with white hairs and 50 with black hairs were enrolled into each radiation group. Stratified analysis showed that the response rates of vitiliginous lesions with white hairs were significantly higher in the perilesional irradiation group (77.6%, 72.8%, respectively) than in the conventional irradiation group before and after the matching (19.3%, 20.7%, respectively, both P < 0.01) ; for the vitiliginous lesions with black hairs, there was no significant difference in the response rate between the 2 groups ( P = 0.908) . Conclusion:The efficacy of NB-UVB irradiation around vitiliginous lesions is superior to the conventional irradiation in the treatment of refractory vitiligo, especially vitiliginous lesions with white hairs.

The multiple-step cleavage of amyloid precursor protein (APP) generates amyloid-β peptides (Aβ), highly toxic molecules causing Alzheimer's disease (AD). The nonspecific cleavage between the transmembrane region of APP (APPTM) and γ-secretase is the key step of Aβ generation. Reconstituting APPTM under physiologically-relevant conditions is crucial to investigate how it interacts with γ-secretase and for future AD drug discovery. Although producing recombinant APPTM was reported before, the large scale purification was hindered by the use of biological protease in the presence of membrane protein. Here, we expressed recombinant APPTM in Escherichia coli using the pMM-LR6 vector and recovered the fusion protein from inclusion bodies. By combining Ni-NTA chromatography, cyanogen bromide cleavage, and reverse phase high performance liquid chromatography (RP-HPLC), isotopically-labeled APPTM was obtained in high yield and high purity. The reconstitution of APPTM into dodecylphosphocholine (DPC) micelle generated mono dispersed 2D ¹⁵N-¹H HSQC spectra in high quality. We successfully established an efficient and reliable method for the expression, purification and reconstruction of APPTM, which may facilitate future investigation of APPTM and its complex in more native like membrane mimetics such as bicelle and nanodiscs.
Humans ; Amyloid beta-Protein Precursor/chemistry* ; Micelles ; Amyloid Precursor Protein Secretases/metabolism* ; Magnetic Resonance Spectroscopy ; Recombinant Proteins

Objective:To fully understand the maternal and neonatal outcomes in pregnant women with COVID-19 and explore the evidence of intrauterine vertical transmission of 2019-nCoV by analyzing clinical and laboratory information in peer-reviewed publications on COVID-19 in pregnant women.Methods:PubMed, Embase, China National Knowledge Infrastructure, China Academic Journals, and Wanfang Databases were searched to retrieve articles on COVID-19 in pregnancy published from December 1, 2019, to April 9, 2020. In addition, the World Health Organization COVID-19 Database and the reference lists in each included article were also searched. All included cases were positive for 2019-nCoV nucleic acid with maternal and neonatal outcomes regardless of delivery or not. Clinical manifestations, perinatal and neonatal outcomes were analyzed systematically.Results:This study reviewed 29 publications involving 146 pregnant women who tested positive for 2019-nCoV nucleic acid and their 116 newborns (including two twins). Five cases of severe COVID-19 and three cases of unidentified type that were admitted to ICU for treatment were severe symptoms, accounting for 5.5% (8/146) of all cases. Totally, 69.9% (102/146) of the women underwent cesarean section and 8.2% (12/146) gave birth vaginally. Thirty (20.5%) women continued their pregnancies. One case (0.7%, 1/146) terminated the pregnancy at 26 weeks of gestation due to bidirectional affective disorder and one (0.7%, 1/146) received artificial abortion at 6 weeks of gestation. Fever (58.2%, 85/146) and cough (32.9%, 48/146) were the most common symptoms. However, 15.8% (23/146) of the pregnant women were asymptomatic on admission and symptoms appeared or became worse after delivery in 20.5% (30/146). Lymphocytopenia (49.6%, 56/113) and elevated C-reactive protein (58.4%, 66/113) were the main laboratory findings. The most common computed tomography (CT) finding was bilateral multiple patchy ground-glass opacity in lungs (79.7%, 94/118). The outcomes of 92.2% (107/116) of the newborns were good, and the rest 7.8% (9/116) showed different abnormalities of varying degrees. Among the nine newborns, six showed different degrees of dyspnea, cyanosis and vomiting including one died of multiple organ failure and disseminated intravascular coagulation; one tested positive for viral nucleic acid 36 hours after birth; one was stillbirth due to unknown reason, but intrauterine vertical transmission was excluded; one neonatal death in a critically ill mother undergoing cesarean delivery.Conclusions:Pregnant women are less likely to progress to severe COVID-19 and mostly have a good outcome. Despite reports of adverse neonatal outcomes, evidence of intrauterine vertical transmission of 2019-nCoV remains insufficient.

OBJECTIVETo study the role of p16 gene mutation status as detected by fluorescence in-situ hybridization (FISH) and p16 protein expression as detected by immunohistochemistry in differential diagnosis of malignant mesothelioma and benign mesothelial hyperplasia.

METHODSp16 gene mutation status and protein expression were detected by FISH and immunohistochemistry respectively in 55 cases of pleural malignant mesothelioma and 30 cases of benign mesothelial hyperplasia.

RESULTSFISH study showed that the rate of p16 deletion in malignant mesothelioma (81.8%,45/55) was higher than that in benign mesothelial hyperplasia (3.3%,1/30). The difference was statistically significant (P<0.05). Immunohistochemical study showed that the rate of p16 protein expression in malignant mesothelioma (23.6%) was lower than that in benign mesothelial hyperplasia (76.7%). The difference was also statistically significant. The sensitivity and specificity of FISH in distinguishing between mesothelioma and reactive mesothelial hyperplasia were higher than those of immunohistochemistry.

CONCLUSIONSIn contrast to reactive mesothelial hyperplasia, p16 gene is deleted and p16 protein is not expressed in malignant mesothelioma. The sensitivity and specificity of FISH are higher than those of immunohistochemistry in the distinction.

Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Diagnosis, Differential ; Epithelium ; pathology ; Genes, p16 ; Humans ; Hyperplasia ; diagnosis ; genetics ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Mesothelioma ; diagnosis ; genetics ; metabolism ; Mutation ; Pleura ; pathology ; Pleural Neoplasms ; diagnosis ; genetics ; metabolism ; Sensitivity and Specificity

G protein-coupled receptors (GPCRs) are involved in all human physiological systems where they are responsible for transducing extracellular signals into cells. GPCRs signal in response to a diverse array of stimuli including light, hormones, and lipids, where these signals affect downstream cascades to impact both health and disease states. Yet, despite their importance as therapeutic targets, detailed molecular structures of only 30 GPCRs have been determined to date. A key challenge to their structure determination is adequate protein expression. Here we report the quantification of protein expression in an insect cell expression system for all 826 human GPCRs using two different fusion constructs. Expression characteristics are analyzed in aggregate and among each of the five distinct subfamilies. These data can be used to identify trends related to GPCR expression between different fusion constructs and between different GPCR families, and to prioritize lead candidates for future structure determination feasibility.
Animals ; Computational Biology ; Crystallography, X-Ray ; Gene Expression ; Humans ; Plasmids ; genetics ; metabolism ; Protein Domains ; Receptors, Adrenergic, beta-1 ; Receptors, G-Protein-Coupled ; classification ; genetics ; metabolism ; Receptors, Odorant ; metabolism ; Receptors, Purinergic P1 ; genetics ; metabolism ; Sf9 Cells ; Spodoptera