Objective To analyze the genomic characteristics of hepatitis A virus(HAV)in environmental sewage in Zhuanghe from 2020 to 2021 and compare them with those in case samples.Methods Eighteen sewage samples(environmental sewage)were collected from the entry point of the sewage treatment plant in Zhuanghe from July to December 2021,and the viral RNA was extracted. After reverse transcription,VP1-2A region was amplified by nested PCR,sequenced and spliced.The five HAV nucleotide sequences(321 bp)obtained were analyzed for phylogenetic tree by using bioinformatics software.Results The HAV of environmental sewage samples in 2021 were all 1A subtype,and the nucleotide sequence homology of VP1-2A region was 98. 7%-100%. The nucleotide sequence homology was 97. 8%-100% with that of the hepatitis A case samples from Zhuanghe in 2020,which could be evolutionarily divided into two branches.Conclusion Subtype 1A was the dominant genotype of HAV in environmental sewage in Zhuanghe,which was different from the genome of the case sample,and also had the same genome of some strains. By comparing the similarities between HAV nucleic acid in environmental sewage and that in case samples,we can better understand the circulation of HAV,so as to make an early warning for the epidemiological monitoring of hepatitis A.

ObjectiveTo find the cognitive function impairment in children with benign epilepsy with centro-temporal spikes and to find a sensitive index which can reveal the cognitive function impairment.Methods21 patients with benign epilepsy with centro-temporal spikes participated in this study. 21 normal subjects, matched for age, gender, years of education and family socioeconomic status, served as controls. Subjects were asked to judged weather the two sequentially presented numbers (S1 and S2) in a digit pair were identical or not, while event-related potential (ERPs) were recorded from electrodes on their scalp.ResultsN270 was evoked by S2 of the conflict condition in either controls or patients. Compared with control group, N270 latency was significantly prolonged and the amplitude decreased in the patient group. However, the peak latency and amplitude of P300 were not changed significantly in patient group. ConclusionChildren with benign epilepsy with centro-temporal spikes as a group show cognitive function impairment. Conflict negative N270 can be employed as a method for evaluating cognitive disturbance, which seems subtler than P300 in detecting mild cognitive function impairment.

Objective:To observe the expression of BTLA on T cells during activation and further analyze its inhibitory effects on T cell activation in different phases.Methods: T cells from PBMC were enriched by negative selection using magnetic beads.Expression of BTLA,CTLA-4 and PD-1 on freshly isolated human T cells and kinetics expression of BTLA,CILA-4 and PD-1 on CD3 mAb stimulated T cells were examined by flow cytometry.T cells were stimulated by anti-CD3 mAb combined with anti-CD28 mAb in the presence of anti-BTLA mAb 8H9,then T cell proliferation was tested by MTT assay in the different culture time.Immature DCs were generated from monocytes cultured in the medium containing GM-CSF and IL-4, and further driven to maturation by anti-CD40 mAb.Expression of HVEM on DCs was measured by flow cytometry.T cells were co-cultured with DCs in the presence of soluble 8H9 or anti-HVEM antibody to block HVEM-BTLA interaction,T cell proliferation was measured by MTT assay.Results:Freshly isolated T cells exhibited high levels of BTLA expression, but not CTLA-4 and PD-1.After T cell activation, BTLA expression decreased on first 2 days, with rapidly increasing to high levels.Unlike BTLA, expression of CTLA-4 and PD-1 was gradually increased during T cell activation.8H9 significantly inhibited the proliferation of T cell stimulated by CD3 mAb and CD28 mAb.8H9 could still exhibit inhibitory effect on T cell proliferation after 24 h or 48 h of preactivation by CD3 mAb plus CD28 mAb stimulation.HVEM was highly expressed on immature DCs, and down-regulated on mature DCs.Blockade of BTLA by soluble 8H9 or anti-HVEM antibody enhanced DC-mediated T cell proliferation within 48 h.Conclusion: BTLA signal enhances the threshold of T cell activation and plays importantly negative regulatory role in the initiation and early phase of T cell activation.

Objective:To evaluate the clinical application of vancomycin in a cancer hospital in order to promote the rational use of vancomycin. Methods:The records of 66 patients treated with vancomycin in 2015 were reviewed and analyzed in the respects of in-dications of medication, pathogenic examination, usage and dosage, course of treatment, drug combination, drug monitoring and drug utilization index(DUI). Results:Among the 66 cases, most drug use was rational, which accounted for 69. 70% with DUI of 0. 82. There were still several types of irrational drug use including no medication indications, irrational usage and dosage, too long or too short course of treatment, irrational combination and prophylactic use time. Conclusion:The application of vancomycin is basically ra-tional, while irrational phenomenon still exists. The management and supervision should be further strengthened in order to promote clinical rational drug use and ensure medication safety of patients.

Objective To study the therapeutic and toxic effects of 32 P-chromic phosphate-poly (L-lactic) acid (32p-CP-PLLA) microparticle intratumoral administration into BALB/c nude mice bearing BxPc-3 human pancreatic carcinoma.Methods Twenty four nude mice bearing tumors were injected with 0,9.3,18.5 and 37.0 M Bq 32p-CP-PLLA microparticle,respectively.The relative tumor growth rates were observed every day,and white blood cells,platelets and body weight were measured.At 14 d after administration,the tumors were removed,histological examination and immunohistochemical analysis were performed.Results The relative tumor growth rates of each treatment group was lower than 40％.Histological examination showed the degenerative necrosis at the site nearby the mircoparticle.Immunohistochemical analysis showed that the Microvessel density (MVD) and the expression of Bcl-2 in treated group were lower than those in control group.In contrast,the expression of bax in treated group were higher than those in control group.The ratio of Bcl-2/Bax protein significantly decreased in the treatment group,which were 3.83 ± 0.43,0.47 ± 0.13,1.10 ± 0.32,2.19 ± 0.57 for 0,9.3,18.5 and 37.0 MBq 32 P-CP-PLLA microparticle,respectively ( t =2.36 - 2.77,P ＜ 0.05).MVD were 31.2 ± 2.3,23.8 ± 1.5,14.8 ±0.8,11.0 ± 1.2,respectively.Dose dependence was observed in both HE and IHC staining after 14 d treatment ( t =2.30 - 2.57,P ＜ 0.05 ).Conclusions Intratumoral injection of 32p-CP-PLLA microparticle might be a safe,easy and effective radionuclide interventional therapy for pancreatic carcinoma.

Objective To study the clinical significance of EGFR mutation in patients with advanced non -small cell lung cancer combined with malignant pleural effusion,and to provide reliable theoretical basis for clinical treatment .Methods 3 0 patients of advanced non -small -cell lung cancer complicated with malignant pleural effusion in Zhoushan island area were selected.DNA was extracted in the pleural effusion and EGFR 19,21 two loci of gene mutation was detected by sequencing PCR.EGFR and clinical characteristics of the patients (gender,age,smoking history,disease types of cases and in the level of CEA level)was compared.Results Among the 30 cases,4 cases of gene mutation,1 case of male patient,3 cases of female patients,4 cases of adenocarcinoma,4 cases of non smokers, 2 cases of EGFR19 deletion,2 cases of EGFR21 mutation.Among them,3 patients were treated by biological target therapy,the survival time was more than 1 year,and there were no obvious adverse reactions,and the effective rate was 75.00%.Conclusion The gene mutations of EGFR were detected in the patients with advanced non -small cell lung cancer combined with malignant pleural effusion,and the mutation rate 13.30%,which was high in female,non smoker and adenocarcinoma,and it could be used to treat the tumor.

Objective To investigate the relative factors of depression in elderly inpatients with multiple chronic diseases. Methods A total of 438 elderly inpatients with multiple chronic diseases were investigated and analyzed by the comprehensive geriatric assessment. Results Among 438 elderly inpatients with chronic diseases, 154 cases(35.15%)were accompanied by depression.The rates of female,non-marital status(single/widowed/divorced),lower income,frequent fall and insomnia were significantly higher in depression group than in non-depression group(P=0.02,0.00,0.00,0.00,0.00,respectively).The scores of cognitive function,nutritional status and activities of daily living were significantly lower in depression group than in non-depression group(P=0.03, 0.00,0.00,respectively),and the pain score was significantly higher in depression group than in non-depression group(P=0.00).The prevalence of diabetes and chronic obstructive pulmonary disease was significantly higher in depression group(P= 0.03,0.04;respectively).Multiple Logistic regression analysis revealed that non-marital status,low income,insomnia,cognitive impairment,malnutrition and diabetes could significantly increase the risk of depression in elderly patients with chronic diseases(OR=2.291,2.065,2.384,2.965,2.561,1.949,respectively,all P<0.05). Conclusions Female,non-marital status,falls,insomnia,cognitive dysfunction,malnutrition,decreased viability of daily life,diabetes,chronic pain and chronic obstructive pulmonary disease are positively associated with the late-life depression.Among them,non-marital status,low income,insomnia,cognitive dysfunction,malnutrition and diabetes could markedly increase the risk of depression in elderly patients with chronic diseases.

ObjectiveTo investigate the effects of aerobic exercise on sleep-deprived rats positioning cruise and space exploration activities, synaptic mechanism, involving hippocampal neuron morphology, dendritic spine density, synaptic related proteins and cyclic adenosine phosphate reactive element binding protein (CREB)/brain-derived neurotrophic factor (BDNF) signaling pathway. MethodsA total of 48 Sprague-Dawley rats were randomly divided into control group (n = 12), model group (n = 12), model-exercise group (n = 12) and exercise group (n = 12). The model-exercise group and the exercise group accepted aerobic exercise with treadmill for four weeks. After exercise intervention, the model group and the model-exercise group were continuously deprived sleep for 72 hours with a multi-platform water environment. They were assessed with Morris Water Maze of positioning cruise and space exploration. The morphology and density of dendritic spines in hippocampal neurons were observed with HE staining and Golgi staining. The expression of hippocampal proteins of postsynaptic density protein 95 (PSD95), synapsin (SYN), growth-associated protein 43 (GAP43), Rac1, BDNF, p-CREB and CREB were measured with Western blotting. ResultsCompared with the control group, the latency and the total swimming distance increased in the model group (P < 0.05); the number of crossing the platform, the target quadrant residence time and distance decreased (P < 0.05); the neurons in the hippocampal CA1 region were disordered, the total density of dendritic spines per unit length and the density of mature, immature and filamentous pseudopodia dendritic spines decreased (P < 0.05); and the expression of PSD95, SYN, GAP43, Rac1, BDNF, p-CREB and CREB decreased (P < 0.05). Compared with the model group, the latency and total swimming distance of the model-exercise group decreased (P < 0.05); the target quadrant residence time and distance increased (P < 0.05); the number of neurons in the hippocampal CA1 region increased and arranged neatly; the total density of dendritic spines per unit length and the density of mature, immature and filamentous pseudopod dendritic spines increased (P < 0.05); and the expression of PSD95, SYN, GAP43, Rac1, BDNF, p-CREB and CREB increased (P < 0.05). ConclusionAerobic exercise can alleviate the learning and memory impairment of sleep-deprived rats, which may relate to the regulation of CREB/BDNF signaling pathway in hippocampus for improvement of synaptic plasticity.

ObjectiveTo investigate the effects of 4 weeks of aerobic exercise combined with traditional Chinese medicine （TCM） on the regulation of phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/glycogen synthase kinase-3β （GSK-3β） signaling pathway and related apoptotic proteins in AD rats， and to explore its mechanism of action in improving cognitive function of AD rats. MethodFifty male SD rats were randomly divided into Sham group， exercise （EX） group， Alzheimer's disease （AD） group （model）， TCM group （Dabuyuan Jian）， and EX+TCM group， 10 in each group. Aβ_25-35 oligomer solution was injected into the hippocampal region to establish the AD rat model， and the EX group underwent 6 days/week of running table training. The TCM group was given Dabuyuan Jian （5.36 g∙kg^-1） by gavage， and the EX+TCM group received running table training combined with intragastric administration of Dabuyuan Jian. An equal volume of purified water was given to the Sham， AD， and EX groups by gavage. The rats' learning memory was evaluated by Morris water maze. The morphological changes and ultrastructural changes of hippocampal neurons were observed by hematoxylin-eosin （HE） staining and transmission electron microscopy， respectively. Western blot was performed to detect the protein expression levels of Akt， p-Akt， GSK-3β， p-GSK-3β， β-catenin， Bcl-2， Bax， and cleaved Caspase-3 in the hippocampus of rats. ResultCompared with the Sham group， the AD group showed an increase in escape latency and total swimming distance， and a decrease in the residence time in the target quadrant （P<0.01）， with loose and disorganized arrangement of hippocampal pyramidal cells， a decrease in density， myelin-like changes in hippocampal ultrastructure， and a blurring of mitochondrial swelling. In addition， the expression of Bcl-2， p-Akt， p-GSK-3β and β-catenin was down-regulated （P<0.05， P<0.01） while the expression of cleaved Caspase-3 was up-regulated （P<0.05）. Compared with the conditions in the AD group， the escape latency and swimming distance were reduced in the EX， TCM， and EX+TCM groups， while the residence time in the target quadrant was increased （P<0.05，P<0.01）， with regularly arranged pyramidal cells， elevated number of cells， and reduced myelin-like changes and improved mitochondrial swelling as seen by transmission electron microscopy. Moreover， the expression of Bcl-2， p-Akt and p-GSK-3β was up-regulated， while the expression of cleaved Caspase-3 was down-regulated（P<0.05，P<0.01）. The EX+TCM group had increased level of β-catenin while the EX and TCM groups were not statistically significant compared with the AD group. Compared with the EX+TCM group， the EX and TCM groups had increased escape latency and swimming distance and decreased residence time in the target quadrant（P<0.05，P<0.01）， with loosely arranged pyramidal cells and reduced synaptic integrity under transmission electron microscopy（P<0.01）. Furthermore， the levels of Bcl-2， p-Akt， p-GSK-3β and β-catenin were lowered （P<0.05，P<0.01）while the level of cleaved Caspase-3 was elevated（P<0.05）. ConclusionAerobic exercise combined with Dabuyuan Jian could improve the cognitive dysfunction of AD rats， and the mechanism might be related to the elevated expression of key proteins in the PI3K/Akt/GSK-3β signaling pathway. The combined use was better than the use of exercise or drugs alone.

Resveratrol (RES), a natural polyphenolic phytochemical, has been suggested as a putative anti-aging molecule for the prevention and treatment of Alzheimer's disease (AD) by the activation of sirtuin 1 (Sirt1/Sir2). In this study, we tested the effects of RES and Sirt1/Sir2 on sleep and courtship memory in a Drosophila model by overexpression of amyloid precursor protein (APP), whose duplications and mutations cause familial AD. We found a mild but significant transcriptional increase of Drosophila Sir2 (dSir2) by RES supplementation for up to 17 days in APP flies, but not for 7 days. RES and dSir2 almost completely reversed the sleep and memory deficits in APP flies. We further demonstrated that dSir2 acts as a sleep promotor in Drosophila neurons. Interestingly, RES increased sleep in the absence of dSir2 in dSir2-null mutants, and RES further enhanced sleep when dSir2 was either overexpressed or knocked down in APP flies. Finally, we showed that Aβ aggregates in APP flies were reduced by RES and dSir2, probably via inhibiting Drosophila β-secretase (dBACE). Our data suggest that RES rescues the APP-induced behavioral deficits and Aβ burden largely, but not exclusively, via dSir2.
Animals ; Alzheimer Disease/metabolism* ; Amyloid beta-Peptides ; Amyloid beta-Protein Precursor/metabolism* ; Drosophila/physiology* ; Drosophila Proteins/metabolism* ; Resveratrol/pharmacology* ; Sirtuin 1 ; Sleep