Objective To determine whether chronic stress could potentiate learning and memory impairment in old mice, and, if so, what the underlying mechanism is. Methods Sixty male mice were divided randomly into control group and chronic stress group. Mice in stress group were stressed everyday by one of the stressors including cold exposure, restraint, level shake and so on. The ability of learning and memory was determined by Morris water maze test, and the histopathologic changes in CA3 field of the hippocampus were examined under a light micro-scope. Serum corticosterone level was determined by enzyme-linked immunosorbent assay. Western blot was per-formed to determine the expression of β-site amyloid precursor protein-cleaving enzyme 1 and Aβ1-42 in hippocam-pus of the brain. Results Compared with the control group, the results showed that chronic stress could increase the escape latency and swimming distance of old mice during training session in the Morris water maze test. The neuropathological changes were characterized by the decreased neuron number,soma shrinkage and condensation,or nuclear pyknosis in the CA3 field of hippocampus in the stress group. On the other hand, the expression of Aβ1-42 and BACE1 protein in hippocampus were increased, as well as the serum corticosterone concentration in the stress group. Conclusion Chronic stress can potentiate learning and memory impairment and pathological damage in CA3 field of the hippocampus in old mice, which may be related to chronic stress up-regulated the levels of BACE1 and Aβ1-42 mediated by corticosterone.

Objective:To analyze the characteristics and prognosis of visual field of Leber hereditary optic neuropathy (LHON) with G11778A mutation.Methods:A retrospective clinical study. Twenty-two (44 eyes) of LHON patients diagnosed with G11778A site mutation by mt-DNA examination from May 2008 to February 2018 in Ophthalmology Department of Dongfang Hospital of Beijing University of Chinese Medicine, were enrolled in this study. All patients underwent best corrected visual acuity (BCVA), visual field and optical coherence tomography (OCT). The BCVA examination was performed using the international standard visual acuity chart, which was converted into logarithm of the minimum angle of resolution (logMAR) BCVA for record. The thickness of the retinal nerve fiber layer (RNFL) in the 200μm×200μm annular region 1.73 mm outside the optic disc was measured by OCT. At least 7 visual field examinations were performed within one month before and after 2, 4, 8, 12, 18, 24 and 30 months of the course of disease by using Octopus 101 perimetry. Among 44 eyes, 27 eyes were detected with G2 procedure (G2 group) and 17 eyes were detected with LVC procedure (LVC group). The mean field defect (MD) and mean optical sensitivity (MS) were used as the main outcome indexes. According to the onset age, the patients were further divided into the ≤14 years old group and>14 years old group. There was a significant difference in initial logMAR BCVA between the G2 group and LVC group ( t=4.994, P=0.000), but there was no significant difference in gender ( χ2=1.896, P=0.169) and age ( t=0.337, P=0.708) between the two groups. Independent sample t test was used for comparison between groups, paired t test was used for comparison within groups, and one-way analysis of variance was used for comparison between groups. The statistical data were compared by χ2 test. Results:In the G2 group, the MD value of the subgroup of children (≤14 years old) decreased gradually during the follow-up period, and the MD value since 18 months after onset was significantly lower than the value of 2 months after onset ( t=3.813, 4.590, 5.033; P=0.002, 0.001, 0.000). No obvious visual field index changes were seen in other subgroups ( P>0.05). The central scotoma was the most common type of visual field defect in the early stage, and the diffuse defect was the most common type of visual field defect in the late stage. There was a significant difference in the types of visual field distribution between the early and late stage in G2 group ( χ2=17.414, P=0.015). There was no significant difference in the type of visual field distribution between the early and late stage in LVC group ( χ2=4.541, P=0.474). The MD value in the G2 group remained stable within 8 months after onset, but significantly improved after 18 months after onset ( t=2.100, 3.217, 3.566; P=0.046, 0.003, 0.001). The MS in the LVC group did not significantly improve during follow-up ( P>0.05). The average visual acuity of the G2 group was significantly improved from 12 months ( t=3.039, 3.678, 4.264, 5.078; P=0.008, 0.002, 0.001, 0.000). The visual acuity of the eyes in the G2 group was better than that of the LVC group during all follow-up periods ( P≤0.05). The RNFL thickness of all patients continued to decrease after onset, but the RNFL thickness was significantly higher at 4, 8, 18, 24, 30 months in the G2 group than those in the LVC group ( t=2.471, 2.269, 2.474, 2.509, 2.782; P=0.018, 0.028, 0.017, 0.016, 0.008). Conclusions:The main types of visual field defect of LHON with G11778A mutation are the central scotoma in the early stage, while the diffuse defect and central scotoma are both very common in the later stage. The visual field of LHON patients examined by G2 procedure is significantly improved during the follow-up, as well as the visual acuity improved significantly, and the visual field improvement in younger cases (≤14 years old) is better than that of older cases (>14 years old), but the visual field of the LVC procedure cases did not improve during follow-up.

Objective To observe the effect of atropine in the treatment of bromiderosis with anhydrous alcohol injection. Methods Patients were randomly divided into two groups (A and B): the patients in Group A was injected with both anhydrous alcohol and atropine, and that in Group B was only injected with anhydrous alcohol. The effect of the operation was evaluated at 1, 3 and 6 months after the treatment. Results From August 2004 to January 2008, 72 patients were involved in this study. 37 cases were included in Group A, and 35 patients were included in Group B. The effective rate in the Group A was 83.78 %, and that in the Group B was 82.86 %. There was no statistical difference between these two groups. Conclusion Atropine has no effect on the treatment of axillary bromidrosis with anhydrous alcohol injection and it is, therefore, not necessarily included in the treatment.

Objective Explore the change of IL-1β and IL-18 expression in pulmonary hypertension induced by monocrotaline.Methods Divide the mouses into two groups, control group and experimental group (n=10).Establish rats pulmonary hypertension model induced by monocrotaline.Detect the model by ultrasound, myocardial cells HE dyeing and tunnel test;ELISA was used to detect the serum biological markers NF-κB, COX2, IL-6, IL-1β, TNF-α and NO;Immunohistochemical was used to detect the expression level of IL-1β and IL-18 in the lung tissue;the protein change of NLRP3 in the lung tissue was detected by Western blot.Results Serum biological markers of NF-κB, COX2, IL-6, IL-1β, TNF-α and NO are significantly increased in PAH rats(P<0.05);The expression of IL-1β, IL-18 in the lung tissue increased obviously(P<0.05);The NLRP3 protein expression was significantly higher in experimental group.Conclusions Changes of NLRP3 effect increase expression of IL-1β and IL-18and which may play an important role in pulmonary hypertension induced by monocrotaline.

With redox-sensitive fluorescene probes DCFH-DA and DHR123, the formation of cytosolic and intramitochondrial reactive oxygen species (ROS) inside immature rat cerebellar granule cells during the apoptosis induced by nitric oxide donor S-nitroso-N-acetyl-pennicillamine (SNAP) was monitored by laser confocal scanning microscopy. The cytosolic and intramitochondrial ROS increase significantly after 0.5 mmol/L SNAP treatment for 1 h. Pre-treatment with the nitric oxide scavenger hemoglobin can effectively inhibit the formation of cytosolic and intrarnitochondrial ROS and protect neurons from apoptosis. Adding glutathione can also protect neurons from apoptosis, and the cytotoxity of nitric oxide increases significantly while the synthesis of glutathione is inhibited. The results indicated that ROS might be involved in NO-induced apoptosis in neural cells and glutathione might be the endogenesis antioxidant to protect neurons from oxidative injury.

Population pharmacokinetics of vancomycin (VAN) in the Chinese patients was described by using nonlinear mixed-effects modeling (NONMEM). 619 VAN serum concentrations data from 260 patients including 177 males and 83 females were collected separately from two centers. A one-compartment model was used to describe this sparse data. No significant difference was observed between two center datasets by introducing SID covariate. The final model was as CL= (θ (base0+ θ(max) x(1 -e(-θ(Age)(Age/72) and V = θ x θ (Age)(Age/72). The creatinine clearance (CL(Cr)) and Age were identified as the most significant covariate in the final model. Typical values of clearance (CL) and volume of distribution (V) in the final model were 2.91 L x h(-1) and 54.76 L, respectively. Internal model validation by Bootstrap and NPDE were performed to evaluate the robustness and prediction of the final model. The median and 95% confidence intervals for the final model parameters were based on 1000 Bootstraps. External model evaluation was conducted using an independent dataset that consisted of 34 patients to predict model performance. Pharmacodynamic assessment for VAN by AUC (0-24 h) to MIC ratios of over 400 was considered to be the best to predict treatment outcomes for patients. AUC (0-24 h) was calculated by clearance based on the above population model. The results indicate that the conventional dosing regimen probably being suboptimal concentrations in aged patients. The approach via population pharmacokinetic of VAN combined with the relationship of MIC, Age, CL(Cr) and AUC(0-24 h)/MIC can predict the rational dose for attaining efficacy.

The insulin receptor (IR) is an important hub in insulin signaling and its activation is tightly regulated. Upon insulin stimulation, IR is activated through autophosphorylation, and consequently phosphorylates several insulin receptor substrate (IRS) proteins, including IRS1-6, Shc and Gab1. Certain adipokines have also been found to activate IR. On the contrary, PTP, Grb and SOCS proteins, which are responsible for the negative regulation of IR, are characterized as IR inhibitors. Additionally, many other proteins have been identified as IR substrates and participate in the insulin signaling pathway. To provide a more comprehensive understanding of the signals mediated through IR, we reviewed the upstream and downstream signal molecules of IR, summarized the positive and negative modulators of IR, and discussed the IR substrates and interacting adaptor proteins. We propose that the molecular events associated with IR should be integrated to obtain a better understanding of the insulin signaling pathway and diabetes.
Animals ; Humans ; Protein Binding ; Receptor, Insulin ; metabolism ; Signal Transduction ; Substrate Specificity

The activation and deactivation of Ca(2+)- and calmodulindependent neuronal nitric oxide synthase (nNOS) in the central nervous system must be tightly controlled to prevent excessive nitric oxide (NO) generation. Considering plasma membrane calcium ATPase (PMCA) is a key deactivator of nNOS, the present investigation aims to determine the key events involved in nNOS deactivation of by PMCA in living cells to maintain its cellular context. Using time-resolved Förster resonance energy transfer (FRET), we determined the occurrence of Ca(2+)-induced protein-protein interactions between plasma membrane calcium ATPase 4b (PMCA4b) and nNOS in living cells. PMCA activation significantly decreased the intracellular Ca(2+) concentrations ([Ca(2+)]i), which deactivates nNOS and slowdowns NO synthesis. Under the basal [Ca(2+)]i caused by PMCA activation, no protein-protein interactions were observed between PMCA4b and nNOS. Furthermore, both the PDZ domain of nNOS and the PDZ-binding motif of PMCA4b were essential for the protein-protein interaction. The involvement of lipid raft microdomains on the activity of PMCA4b and nNOS was also investigated. Unlike other PMCA isoforms, PMCA4 was relatively more concentrated in the raft fractions. Disruption of lipid rafts altered the intracellular localization of PMCA4b and affected the interaction between PMCA4b and nNOS, which suggest that the unique lipid raft distribution of PMCA4 may be responsible for its regulation of nNOS activity. In summary, lipid rafts may act as platforms for the PMCA4b regulation of nNOS activity and the transient tethering of nNOS to PMCA4b is responsible for rapid nNOS deactivation.
Animals ; Brain ; metabolism ; Calcium ; metabolism ; Cells, Cultured ; Cerebellum ; cytology ; Fluorescence Resonance Energy Transfer ; HEK293 Cells ; Humans ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type I ; metabolism ; PDZ Domains ; Plasma Membrane Calcium-Transporting ATPases ; metabolism ; Protein Interaction Maps ; Protein Isoforms ; metabolism ; Rats ; Rats, Sprague-Dawley

Apoptosis ; Calcineurin ; metabolism ; Chymotrypsin ; metabolism ; Humans ; Lysosomes ; enzymology ; Mitochondria ; metabolism ; pathology ; Mitochondrial Dynamics ; Neuroblastoma ; metabolism ; pathology

Objective:To evaluate the status of T, B and NK lymphocytes in peripheral blood of patients with chronic hepatitis B virus(HBV) infection and low-level viremia after nucleos(t)ide analogue (NA) treatment and to provide ideas for solving low-level viremia.Methods:This retrospective study involved 344 patients with chronic HBV infection who had been treated with NAs. They were divided into two groups: low-level viremia group (LLV group) and complete virological response group (CVR group). Clinical data including basic information, biochemistry and coagulation test results, HBV DNA, peripheral blood lymphocyte counts, PD1 and CD28 expression by T lymphocytes, and perforin and granzyme B expression by NK lymphocytes were collected and compared between the two groups. Propensity matching analysis was performed to verify the accuracy of the results.Results:Among the 344 cases, 162 were in the LLV group and 182 in the CVR group. There were no significant differences in disease diagnosis, alanine aminotransferase (ALT), aspartate aminotransferase (AST) or albumin (ALB) level between the two groups ( P>0.05), but the differences in gender and age were statistically significant ( P<0.05). The differences in the counts and percentages of peripheral blood CD3 +, CD4 + and CD8 + T lymphocyte and CD4 + /CD8 + ratios between the two groups were not statistically significant ( P>0.05), but the expression of PD1 and CD28 by peripheral blood CD3 +, CD4 + and CD8 + T lymphocytes was higher in the LLV group than in the CVR group ( P<0.05). The count of peripheral blood CD19 + B lymphocytes in the LLV group was higher than that in the CVR group ( P>0.05), and the percentage of peripheral blood CD19 + B lymphocytes was also higher in the LLV group ( P<0.05). The count of peripheral blood CD16 + CD56 + NK lymphocytes and the expression of perforin in the LLV group were lower than those in the CVR group ( P>0.05). The percentage of peripheral blood CD16 + CD56 + NK lymphocytes and the expression of granzyme B in the LLV group were lower than those in the CVR group ( P<0.05). After propensity score matching, 108 cases in the LLV group and 108 cases in the CVR group showed no significant differences in basic information ( P>0.05); the percentage of CD4 + T lymphocytes and CD4 + /CD8 + ratio in peripheral blood T lymphocyte subsets were higher in the LLV group than in the CVR group, while the percentage of CD8 + lymphocytes was lower in the LLV group ( P<0.05); the expression of PD1 and CD28 by CD3 +, CD4 + and CD8 + T lymphocytes remained higher in the LLV group ( P<0.05); the differences in the counts and percentages of peripheral blood CD19 + B lymphocytes as well as CD16 + CD56 + NK lymphocytes between the two groups were not statistically significant ( P>0.05); no significant difference in the expression of perforin by CD16 + CD56 + NK lymphocytes was found between the two groups ( P>0.05), and the expression of granzyme B remained lower in the LLV group ( P<0.05). Conclusions:Abnormal number and function of T lymphocytes and decreased function of NK lymphocytes might be related to the development of LLV in patients with chronic HBV infection after treatment. Therefore, in addition to adjusting NAs, targeting of T and NK lymphocytes might also be a feasible measure for future LLV treatment.