Objective To investigate the effect of rapid eye movement (REM) sleep deprivation on spatial memory and hippocampal cellular prion protein (PrPC) expression and to explore the underlying mechanism of cognitive impairment induced by sleep deprivation.Methods Adult Sprague-Dawley rats were sorted by weight,randomly divided into three groups:the cage control (CC) group,the tank control (TC) group,and the sleep deprivation (SD) group.Rats were deprived of REM sleep for 72 h using the modified multiple platform method.The Morris water maze task was used to assess hippocampal-dependent spatial memory.After sleep deprivation,the rats were sacrificed and their brain tissue was analyzed for PrPC protein expression via Western blotting.Hippocampal neuron axon elongation was examined as well after lentivector-mediated RNA interference (RNAi) of PrPC in primary cultured rat hippocampal neurons.Results REM sleep deprivation for 72 h resulted in spatial memory impairment.The number of times of rats passing through the platform was decreased significantly in the SD group (3.17 ±0.95) compared with the CC (7.17 ±0.95) and TC (6.50 ±0.62) groups (Z =2.026 6,Z =2.026 6,P ＜0.05),the mean value of proximity to the platform (mm) was greater for rats of the SD group (711.74 ± 33.99) compared to those of theCC (592.32±31.31) andTC (580.86±11.36) groups (Z=-2.001 6,Z=-2.4820,P ＜ 0.05).REM sleep deprivation for 72 h resulted in reduced PrPC level in the hippocampus (0.33 ± 0.10) compared with the CC (1.01 ±0.33) and TC (0.96 ±0.27) groups (Z=2.152 9,Z=2.152 9,P ＜ 0.05).In primary cultured hippocampal neurons,axon elongation(μm) was inhibited 7 days in infected neurons (326.28 ± 12.53) compared with normal (555.00 ±30.43) or negative control (558.70 ±23.10) cells (Z =4.768 4,Z =4.877 0,P ＜ 0.05).Conclusion These findings suggest that PrPC-mediated hippocampal neuron axon elongation inhibition is probably involved in spatial memory impairment induced by sleep deprivation in rats.

This study is to investigate the effects of concentration, intestinal section, pH, paracellular route, substrate/inhibitor of enzyme (CYP3A) and proteins (P-gp, MRP2, SGL1) on the absorption of forsythoside A. The absorption of three concentrations (2.6, 5.2, and 10.4 microg x mL(-1)) of forsythoside A in different intestinal segments was studied with phenol red as the marker by rat circulation in situ. The results showed that the residue of forsythoside A with different concentrations had little significant difference from that obtained after perfusing via duodenum, jejunum, ileum and colon, which indicated that the absorption of forsythoside A was passive diffusion and had no difference in different segments of rat intestine. The residue of forsythoside A increased to 466.160 and 463.429 microg respectively when cyclosporine (4 microg x mL(-1)) or midazolam (50 micromol x L(-1)) was added to the circulation fluid, which showed significant difference compared to the control group (P < 0.05). Moreover, the residue of forsythoside A showed a tendency of increase with the increase of cyclosporine or midazolam. When digoxin (50 micromol x L(-1)) or EDTA (10 microg x mL(-1)) was added to the circulation fluid, the residue of forsythoside A decreased to 325.110 and 369.888 microg respectively, which showed significant difference as compared to the control group (P < 0.05). Besides, the residue of forsythoside A showed a tendency of reduction with the increase of digoxin or EDTA. However, there is no significant change in the absorption of forsythoside A when the different concentrations of mannitol were added to the circulation fluid. The results above indicated that the absorption of forsythoside A was mainly passive diffusion and involved paracellular route at the same time. In addition, the substrates of P-gp or CYP3A had dose-dependent effect on the absorption of forsythoside A.

Objective To introduce the application of multiplex ligation-dependent probe amplification (MLPA) assays in the diagnosis of patients with hereditary neuropathy with liability to pressure palsies (HNPP).Methods Copy numbers of the exons in peripheral myelin prolein 22 (PMP22) gene,tektin 3 (TEKT3) gene and cytochrome c oxidase assembly protein 10 (COX10) gene were analyzed by MLPA in 8 patients diagnosed with HNPP clinically and 5 normal controls.Results Among the 8 patients,7 patients were identified to have deletion mutations according to their reduced peak area of PMP22 gene,TEKT3 gene and COX10 gene compared with that of normal controls.One patient with normal peak area of PMP22 gene,TEKT3 gene and COX10 gene showed no deletion of these genes.Conclusions MLPA assays can detect the copy numbers of genes in HNPP region through semi-quantitative analysis in a rapid,accurate way,which may be utilized widely in the genetic diagnosis among HNPP patients.

Objective To investigate the association of genetic polymorphisms of ATIC and GSTP1 with plasma concentrations and adverse reactions of high-dose methotrexate( HD-MTX)in children with acute lymphoblastic leukemia (ALL). Methods A total of 70 peripheral blood samples were obtained from ALL children for extraction of genome DNA.The gene polymorphisms of ATIC T26293C and GSTP1 A313G locus were examined by using PCR and direct sequencing.Enzyme multiplied immunoassay technique(EMIT)was employed to determine the plasma concentration of MTX in 48 h.Clinical data of patients were collected during HD-MTX chemotherapy,and the adverse reactions were statistically analyzed.The associations of ATIC and GSTP1 genotypes with MTX plasma concentration and adverse reactions were investigated. Results There were genetic polymorphisms at the SNP of ATIC T26293C and GSTP1 A313G.At the SNP of ATIC T26293C,the percentages of TT, CT and CC genotypes in ALL children were 4.35%,39.13% and 56.52%,respectively,and the frequencies of T and C alleles were 23.91% and 76.09%.At the SNP of GSTP1 A313G,the percentages of AA,GA and GG genotype were 68.57%,28.57%and 2.86%,respectively,in ALL children. The frequencies of A and G alleles were 82. 86% and 17. 14%,respectively. No statistically significant difference was found in the ratio of blood MTX concentration to MTX dose at 48 h between children with different genotypes(P>0.05).In the GSTP1 A313G site,genotypes that induced the gastrointestinal reactions in the order from low to high were AA,GA,GG,and there was a significant association between gene polymorphism and gastrointestinal side effects(P<0.05).In the GSTP1 A313G site,genotypes that induced myelosuppression in the order of low to high were GG,AA, GA,and a significant association was noted between gene polymorphism and myelosuppression(P<0.05). Conclusion There are significant associations between GSTP1 A313G polymorphism and gastrointestinal side effects or myelosuppression after HD-MTX chemotherapy in ALL children.

Objective:To investigate epidemiological characteristics of outpatients and disease spectrum in the dermatology department during the COVID-19 epidemicMethods:A retrospective comparison of outpatient visits, gender, age and disease types in the dermatology department of Wuhan No.1 Hospital was performed between COVID-19 epidemic period (from 23th January 2020 to 15th April 2020) and the same period in 2019. Enumeration data were analyzed by Pearson′s chi-square test.Results:During the COVID-19 epidemic, the number of outpatient visits to the dermatology department of the hospital decreased markedly, and the average daily number of outpatient visits (236 visits/day) was only 8.81% of that during the same period in 2019 (2 678 visits/day) ; the ratio of male to female patients was reversed from 1∶1.37 in 2019 to 1.16∶1 in 2020; the proportions of patients aged 0-6, 7-12, 13-17 and 18-45 years significantly decreased compared with those in 2019 (all P < 0.001) , and the proportions of patients aged 46-69 and > 69 years significantly increased (both P < 0.001) . During the COVID-19 epidemic, there were 171 types of skin diseases in the dermatology outpatient department, and the number of disease categories decreased compared with that during the same period in 2019 (442 types) ; the number of patient visits for allergic skin diseases, erythematous papulosquamous skin diseases, viral infectious skin diseases and bacterial infectious skin diseases significantly increased compared with that during the same period in 2019 (all P < 0.001) , while the number of patient visits for sebaceous and sweat gland disorders, pigmented skin diseases and physical skin diseases significantly decreased (all P < 0.001) . Conclusion:Compared with the same period in 2019, the number of outpatient visits, patient sex ratio, age distribution and disease types in the dermatology department have undergone marked changes during the COVID-19 epidemic, and this study provides a reference for healthcare workers in dermatology department to respond to various epidemics and natural disasters in the future.

OBJECTIVETo research the influence of glycyrrhiza extract on the pharmacokinetics characteristic parameters of daphnetin, which was aimed to explore the rationality of concert application of drugs.

METHODThe rats received intragastric administration of daphnetin and glycyrrhiza extract containing the same daphnetin respectively. The blood concentration of daphnetin was assayed by LC-MS. The data was processed by program DAS2.1.1.

RESULTGlycyrrhiza extract can reduce the t(1/2), tmax and Ke of daphnetin, while increased the Ka and AUC(0-infinity).

CONCLUSIONGlycyrrhiza extract promoted the oral absorption of daphnetin, slowed down the elimination and increased the biological availability.

Animals ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacology ; Glycyrrhiza ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tissue Distribution ; drug effects ; Umbelliferones ; pharmacokinetics

OBJECTIVETo study the intestinal absorption kinetics characteristic of the main four active ingredients in Shuanghuanglian oral liquid (SHL) and to investigate the influence of herbal compatibility in SHL on absorption of main effective ingredients.

METHODThe in situ rat circulation model was used to investigate the concentration change differences of the four active components in SHL during perfusion.

RESULTThe absorption quantity of different concentrations of baicalin, chlorogenic acid, phillyrin and forsythoside A ranging from 40-160, 6-24, 3-12, 2.6-10.4 mg x L(-1) respectively was linear with concentration and showed no saturation at high concentration. The absorption rate constant K(a) and the hourly absorption percentages A were essentially unchanged. The pH changing from 5.0-7.43 had little influence on the absorption of phillyrin except baicalin, chlorogenic acid and forsythoside A. The calculated K(a) and A of the four active ingredients had no significant differences from that obtained after perfusing via duodenum, jejunum, ileum and colon; The calculated K(a) and A of baicalin in Scutellariae Radix (SR), chlorogenic acid in Lonicerae Japonicae Flos (LJF) and phillyrin in Forsythiae Fructus (FF) had no significant differences compared with that in SHL, but the calculated K(a) and A of forsythoside A in FF were obviously superior to that in SHL.

CONCLUSIONThe intestinal absorption of the four active ingredients in SHL was mainly passive diffusion and had no difference in different segments of rat intestine; the compatibility of SHL compounds changed the absorption of forsythoside A in FF obviously.

Administration, Oral ; Animals ; Chlorogenic Acid ; analysis ; pharmacokinetics ; Drugs, Chinese Herbal ; chemistry ; Flavonoids ; analysis ; pharmacokinetics ; Glucosides ; analysis ; pharmacokinetics ; Glycosides ; analysis ; pharmacokinetics ; Hydrogen-Ion Concentration ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Objective:To investigate the effect of gallic acid on the morphology, proliferation and cell cycle of keloid fibroblasts, as well as on collagen contraction and the transforming growth factor-β (TGF-β) /Sma- and Mad-related proteins (Smads) signaling pathway, and to explore the role and mechanisms of action of gallic acid in the treatment of keloids.Methods:From August to December 2022, 3 keloid tissue samples were collected from 3 patients with clinically and pathologically confirmed keloids after surgery in the Department of Dermatologic Surgery, Wuhan No.1 Hospital. Primary fibroblasts were isolated and cultured by using the tissue culture method, and 3- to 8-passage fibroblasts were used for subsequent experiments. Cultured keloid fibroblasts were divided into 4 groups: low-, medium- and high-dose gallic acid groups treated with 0.025, 0.05 and 0.1 mg/ml gallic acid respectively, and a control group cultured with Dulbecco′s modified Eagle′s medium (DMEM) containing 10% fetal calf serum. After 24-, 48-, and 72-hour treatment, cellular proliferative activity was evaluated by cell counting kit 8 (CCK8) assay, and collagen contraction by using a three-dimensional culture method. After 24-hour treatment in the above groups, pictures were taken using a differential interference inverted fluorescence microscope, and changes in the cell cycle were analyzed by flow cytometry. Some keloid fibroblasts were divided into 2 groups: an experimental group (high-dose gallic acid group) treated with 0.1 mg/ml gallic acid, and a control group cultured with DMEM containing 10% fetal calf serum. After 24-hour treatment, enzyme-linked immunosorbent assay (ELISA) was performed to determine the changes in supernatant concentrations of TGF-β1, TGF-β2, and TGF-β3 in the two groups, real-time fluorescence-based quantitative PCR to detect the relative mRNA expression levels of TGF-β1, TGF-β2, TGF-β3, Smad2, Smad3, Smad4, and α-smooth muscle actin (α-SMA). Statistical analysis was carried out using t test, one-way analysis of variance and two-way analysis of variance, and least significant difference (LSD) - t test was used for multiple comparisons. Results:Compared with the control group, the gallic acid groups showed gradual changes in the shape of keloid fibroblasts under the microscope as the dose of gallic acid increased, including gradually shrinking cell bodies, enlarged intercellular spaces, cell atrophy, increased number of apoptotic cells, etc. CCK8 assay showed that the cellular proliferative activity changed significantly as the dose of gallic acid increased and the treatment time was prolonged ( Fgroup = 78.31, P < 0.001; Ftime = 4.17, P = 0.037), and the proliferative activity of keloid fibroblasts was significantly lower in the high-dose gallic acid group than in the control group at 24, 48, and 72 hours (all P < 0.05). The three-dimensional culture showed that different degrees of collagen contraction occurred in all groups over time, marked collagen contraction was observed in the control group, and a lower degree of collagen contraction in the gallic acid groups; the collagen contraction indices were significantly lower in the medium- and high-dose gallic acid groups than in the control group at 24, 48, and 72 hours (all P < 0.05). Flow cytometry showed that the cell apoptosis rates were significantly higher in the low-, medium- and high-dose gallic acid groups (38.68% ± 3.05%, 41.82% ± 2.19%, 43.56% ± 3.58%, respectively) than in the control group (12.58% ± 1.56%, all P < 0.001) after 24-hour treatment; compared with the control group, the medium- and high-dose gallic acid groups showed significantly decreased proportions of cells in the G0/G1 phase (both P < 0.01), but significantly increased proportions of cells in the S phase and G2/M phase (all P < 0.05). ELISA revealed that the TGF-β1 concentration was significantly lower in the high-dose gallic acid group (758.58 ± 31.42 pg/ml) than in the control group (1 081.30 ± 44.72 pg/ml, t = 11.81, P<0.001), there was no significant difference in the TGF-β2 concentration between the high-dose gallic acid group (71.05 ± 7.40 pg/ml) and the control group (76.43 ± 6.51 pg/ml, t = 1.09, P = 0.317), while the TGF-β3 concentration was significantly higher in the high-dose gallic acid group (5.70 ± 3.87 pg/ml) than in the control group (0.00 ± 0.00 pg/ml, t = 2.94, P = 0.026). As real-time fluorescence-based quantitative PCR revealed, the high-dose gallic acid group showed significantly decreased mRNA expression levels of TGF-β1, Smad2, Smad3, Smad4, and α-SMA (all P < 0.05), but significantly increased mRNA expression level of TGF-β3 ( t = 6.78, P = 0.002) compared with the control group; however, there was no significant difference in the TGF-β2 mRNA expression level between the above two groups ( t = 0.05, P = 0.962) . Conclusion:Gallic acid could change the cell cycle, inhibit the proliferative activity, promote apoptosis and change the shape of keloid fibroblasts, and thus inhibit scar formation and contraction, which may be related to the inhibition of TGF-β/Smads signaling pathway.

Objective:To explore the effects of Rougan Huaxian Granules combined with nucleoside antiviral drugs on liver and kidney function, portal hemodynamics, vascular activity, antiviral indexes and aspartate transaminase-platelet ratio index in patients with hepatitis B decompensated cirrhosis.Methods:A case-control study was conducted on 150 patients with hepatitis B decompensated cirrhosis who were hospitalized in Tangshan Infectious Disease Institute and Affiliated Hospital of North China University of Science and Technology from June 2017 to December 2019 were enrolled. The patients were divided into control group and observation group by computer random random number method, with 75 cases in each group. The control group was given routine liver protection and antiviral treatment; the observation group was given Rougan Huaxian granules on the basis of the control group treatment. Observe the changes of liver and kidney function, portal vein system hemodynamics, vascular activity, antiviral index and aspartate transaminase-platelet ratio index in the two groups. Independent sample T test was used to compare the measurement data between the two groups, paired T test was used for comparison between the same groups before and after treatment, and χ2 test was used for counting data. Results:There were no significant differences in gender, age, course of cirrhosis, Child grade of liver function and baseline data of indexes before treatment between 2 groups (ALL P>0.05). After treatment, alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen, creatinine,diameter of portal vein (Dpv), diameter of splenic vein (Dsv), endothelin-1, nitric oxide, glucagon (GLA), APRI,were all lower than before treatment. Comparison between groups, observation group ALT (51.60±15.97) U/L, AST (62.65±26.28) U/L, urea nitrogen (10.25±1.65) mmol/L, creatinine (78.54±14.09) μmol/L, Dpv (10.20±1.10) mm, Dsv (8.08±0.68) mm, endothelin-1 (31.93±6.35) ng/L, nitric oxide (41.38±8.06) μg/L, GLA (69.54±12.14) mg/L, APRI (3.14±1.35), were significantly lower than those of control group ((97.49±30.87) U/L, (96.03±25.63) U/L, (17.49±2.55) mmol/L, (116.43±22.77) μmol/L, (13.42±1.26) mm, (10.44±0.83) mm, (44.34+11.88) ng/L, (63.47±15.50) μg/L, (107.11+25.29) mg/L, (5.91±1.93)), the differences were statistically significant ( t values were respectively 11.43, 7.87, 20.64, 12.26, 16.62, 18.99, 7.98, 10.96, 11.60, 10.23, all P<0.05). After treatment, albumin, portal vein velocity (Vpv), and velocity of splenic vein blood flow (Vsv) were all higher in the two groups than before treatment. However, there was no significant difference in Vsv of the control group before and after treatment ( t=0.51, P=0.613). Comparison between groups, albumin (39.42±7.35) g/L, Vpv ((25.72±4.06) cm/s), Vsv ((24.22±6.15) cm/s) in the observation group were significantly higher than those in the control group (34.66±7.95) g/L, (19.38±3.46) cm/s, (19.54±5.88) cm/s ( t values were 3.81, 10.28, 4.76, all P<0.05). After treatment, the total effective rate (96.00%(72/75) vs. 86.67%(65/75), χ2=4.13, P=0.042), HBV DNA negative conversion rate (76.00%(57/75) vs. 58.67%(44/75), χ2=5.12, P=0.024), HBeAg negative conversion rate (50.67%(38/75) vs. 30.67%(23/75), χ2=6.22, P=0.013) and serum HBeAg/HBeAb conversion (28.00%(21/75) vs. 13.33%(10/75), χ2=4.92, P=0.027) in observation group were higher than those in control group, and the differences were statistically significant ( P<0.05). HBsAg negative rate (8.00%(6/75) vs. 5.33%(4/75), χ2=0.43, P=0.513) was higher than that of control group, but the difference was not statistically significant ( P>0.05). Conclusion:Rougan Huaxian Granules combined with nucleoside antiviral drugs has significant effect on patients with decompensated liver cirrhosis of hepatitis B, improve liver and kidney function, liver fibrosis and hemodynamics of the portal vein system, increase vascular activity function, and reduce hepatitis B virus (HBV) DNA load, HBV replication, aspartate transaminase-platelet ratio index, APRI, Toll-like receptor (TLR-4) and transforming growth factor β1 (TGF-β1) levels and improves the body′s immune status.

Objective:To explore the effects of embodied emotion priming on attentional bias of individuals with depression tendency.Methods:From June to December 2018, a total of 91 college students with depression tendency were recruited to participate in the experiment.A 3(embodied emotion priming: positive priming, negative priming and no priming) × 2 (emotional face: happy and sad) mixed design was adopted to measure the attentional bias of individuals with depression tendency using the dot probe paradigm. SPSS 22.0 statistical software was used for repeated measurement analysis of variance.Results:In terms of attentional bias, the interaction effect between embodied emotion priming types and emotional faces was significant ( F(2, 88)=5.97, P=0.004, ηp2=0.119). Further simple effect analysis showed that, under the happy-face condition, participants' attentional bias reaction time(△RT) was significantly higher when primed with embodied positive emotion than those primed with embodied negative emotion((14.30±18.23)ms, (-6.53±38.17)ms, P<0.05). The participants' attentional bias △RT was significantly lower when primed with embodied negative emotion than participants with no priming ((-6.53±38.17)ms, (9.16±30.62)ms, P<0.05). Under the sad-face condition, the participants' attentional bias △RT was significantly higher when primed with embodied negative emotion((28.22±35.33)ms) than participants primed with embodied positive emotion((11.71±29.24)ms, P<0.05) and no priming ((7.63±30.60)ms, P<0.05). Conclusion:Embodied emotion priming can affect the attentional bias of individuals with depression tendency.