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A growing number of studies have identified sex differences in response to general anesthesia; however, the underlying neural mechanisms are unclear. The medial preoptic area (MPA), an important sexually dimorphic structure and a critical hub for regulating consciousness transition, is enriched with estrogen receptor alpha (ERα), particularly in neuronal clusters that participate in regulating sleep. We found that male mice were more sensitive to sevoflurane. Pharmacological inhibition of ERα in the MPA abolished the sex differences in sevoflurane anesthesia, in particular by extending the induction time and facilitating emergence in males but not in females. Suppression of ERα in vitro inhibited GABAergic and glutamatergic neurons of the MPA in males but not in females. Furthermore, ERα knockdown in GABAergic neurons of the male MPA was sufficient to eliminate sex differences during sevoflurane anesthesia. Collectively, MPA ERα positively regulates the activity of MPA GABAergic neurons in males but not in females, which contributes to the sex difference of mice in sevoflurane anesthesia.
Subject(s)
Animals , Female , Male , Mice , Anesthesia , Estrogen Receptor alpha/metabolism , Preoptic Area , Sevoflurane/pharmacology , Sex CharacteristicsABSTRACT
The lateral hypothalamic area (LHA) plays a pivotal role in regulating consciousness transition, in which orexinergic neurons, GABAergic neurons, and melanin-concentrating hormone neurons are involved. Glutamatergic neurons have a large population in the LHA, but their anesthesia-related effect has not been explored. Here, we found that genetic ablation of LHA glutamatergic neurons shortened the induction time and prolonged the recovery time of isoflurane anesthesia in mice. In contrast, chemogenetic activation of LHA glutamatergic neurons increased the time to anesthesia and decreased the time to recovery. Optogenetic activation of LHA glutamatergic neurons during the maintenance of anesthesia reduced the burst suppression pattern of the electroencephalogram (EEG) and shifted EEG features to an arousal pattern. Photostimulation of LHA glutamatergic projections to the lateral habenula (LHb) also facilitated the emergence from anesthesia and the transition of anesthesia depth to a lighter level. Collectively, LHA glutamatergic neurons and their projections to the LHb regulate anesthetic potency and EEG features.
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OBJECTIVE@#To develop a fast, sensitive and cost-effective method based on resonance light scattering (RLS) for characterization of protein solubility to facilitate detection of changes in solubility of mutant proteins.@*METHODS@#We examined the response curve of RLS intensities to the protein concentrations in synchronous scanning mode. The curve intersection points were searched to predict the maximal concentrations of the protein in dispersion state, which defined the solubility of the protein in this given state. Bovine serum albumin (BSA, 0-50 g/L) was used as the model to investigate the influences of pH values (6.5, 7.0, and 7.4) and salt concentrations (0.05, 0.10, 0.15, and 0.20 mol/L) on the determined solubility. The solubility of glutathione S-transferase isoenzymes alpha (GSTA, 0-27.0 g/L) and Mμ (GSTM, 0-20.0 g/L) were estimated for comparison. The RLS-based method was used to determine the solubility of uricase (MGU, 0-0.4 g/L) to provide assistance in improving the solubility of its mutants.@*RESULTS@#We identified two intersection points in the RLS response curves of the tested proteins, among which the lower one represented an approximation of the maximal concentration (or the solubility of the protein) in single molecular dispersion, and the higher one the saturated concentration of the protein in multiple molecular aggregation. In HEPES buffer, the two intersection points of BSA (isoelectric point 4.6) both increased with the increase of pH (6.5-7.4), and their values were ~1.2 g/L and ~33 g/L at pH 7.4, respectively; the latter concentration approached the solubility of commercial BSA in the same buffer at the same pH. The addition of NaCl reduced the values of the two intersection points, and increasing salt ion concentration decreased the values of the lower intersection points. Further characterizations of GSTA and GSTM showed that the low concentration intersection points of the two proteins were ~0.7 g/L and ~0.8 g/L, and their high concentration intersection points were ~10 g/L and ~11 g/L, respectively, both lower than those of BSA, indicating the feasibility of the direct characterization of protein solubility by RLS. The two concentration intersection points of MGU were 0.24 g/L and 0.30 g/L, respectively, and the low concentration intersection point of its selected mutant was increased by 2 times.@*CONCLUSIONS@#RLS allows direct characterization of the solubility of macromolecular proteins. This method, which is simple and sensitive and needs only a small amount of proteins, has a unique advantage for rapid comparison of solubility of low-abundance protein mutants.
Subject(s)
Hydrogen-Ion Concentration , Light , Scattering, Radiation , Solubility , Spectrum AnalysisABSTRACT
As a receptor tyrosine kinase, mesenchymal to epithelial transition factor (MET) is the membrane receptor for hepatocyte growth factor (HGF), which is related with a series of biological functions, such as cell proliferation, progression, apoptosis, metastasis and morphological changes. As research continues, MET is amplified or overexpressed in a wide range of human cancers and closely related with worse prognosis. Therefore, various MET inhibitors are currently being developed as potential treatments for a variety of cancers. Based on our current study we summarize the existing knowledge on structure, biological function and its inhibitors of MET and provide a data phase for future researchers.
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Objective To investigate the association of human papilloma virus (HPV) typing and p53 expression with radiosensitivity in patients with cervical cancer.Methods A total of 80 patients with cervical cancer from 2014 to 2016 were enrolled,and among these patients,40 had stage Ⅰ B+ Ⅱ A disease and 40 had stage Ⅱ B +ⅢA disease.HPV genotype was identified and p53 expression was measured.All the patients underwent external and internal pelvic irradiation alone,and the correlation between short-term therapeutic effect and HPV typing/p53 expression was analyzed.The chi-square test or the Fisher's exact test was used for statistical analysis,and Spearman rank correlation analysis was also performed.Results The radiotherapy-insensitive (stable disease+progressive disease) group had higher p53 positive rates than the radiotherapy-sensitive (complete response+partial response) group (stage Ⅰ B+ Ⅱ:100％ vs.80.0％,P=0.044;stage Ⅱ B+ⅢA:100％ vs.90.0％,P=0.013).The expression of p53 was negatively correlated with radiosensitivity (r =-0.427,P =0.000).In the radiotherapy-insensitive group of patients with stage I B + Ⅱ and Ⅱ B +Ⅲ A,the rate of HPV multiple infections was higher than that of single subtype infection (65.0％/95.0％ vs.35.0％/5.0％,P=0.004 and 0.003),while in the radiotherapy-sensitive group,the rate of single subtype infection was higher than that of multiple infections (85.0％/60.0％ vs.15.0％/40.0％,P=0.004 and 0.003).The highest detection rate of HPV16 was 66.3％ in all patients,and the highest detection rate of HPV18 was 60.0％ in the radiotherapy-insensitive group.Conclusions High expression of p53 is associated with radioresistance in patients with cervical cancer.Patients with HPV muhiple infections have poor radiosensitivity,and HPV16 is the most common subtype in dual infection.Among patients who do not achieve remission after radiotherapy,HPV multiple infections with HPV18 as the main pathogen has the highest detection rate.
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Objective To investigate the mutations of BRCA genes in sporadic high grade serous ovarian cancer (HGSOC) and study its clinical significance. Methods Sixty-eight patients between January 2015 and January 2016 from the Affiliated Cancer Hospital of Zhengzhou University were collected who were based on pathological diagnosis of ovarian cancer and had no reported family history, and all patients firstly hospitalized were untreated in other hospitals before. (1)The BRCA genes were detected by next-generation sequencing (NGS) method. (2)The serum tumor markers included carcinoembryonic antigen (CEA), CA125, CA199, and human epididymis protein 4 (HE4) were detected by the chemiluminescence methods, and their correlation was analyzed by Pearson linear correlation. Descriptive statistics and comparisons were performed using two-tailed t-tests, Pearson′s chi square test, Fisher′s exact tests or logistic regression analysis as appropriate to research the clinicopathologic features associated with BRCA mutations, including age, International Federation of Gynecology and Obstetrics(FIGO)stage, platinum-based chemotherapy sensitivity, distant metastases, serum tumor markers (STM). Results (1) Fifteen cases (22%, 15/68) BRCA mutations were identified (BRCA1: 11 cases; BRCA2: 4 cases), and four novel mutations were observed. (2) The levels of CEA, CA199, and HE4 were lower in BRCA mutations compared to that in control group, while no significant differences were found (P>0.05), but the level of CA125 was much higher in BRCA mutation group than that in controls (t=-3.536,P=0.003). Further linear regression analysis found that there was a significant linear correlation between CA125 and HE4 group (r=0.494,P0.05), while significant differences were found in CA125 and sensitivity to platinum-based chemotherapy between the patients with BRCA mutation and wild type (P<0.05). The multiple factors analysis showed that the high level of CA125 was a independent risk factor of BRCA mutations in sporadic HGSOC (P=0.007). Conclusion The combination of CA125 with BRCA have great clinical significance, the mutation of BRCA gene could guild the clinical chemotherapy regiments.