The contents of amino acids in dorsal hippocampus were measured at themoment of 10-min transient forebrain ischemia by using amino acid autoanalyser, delayedneuronal death (DND) model was established by bilateral carotid arteries occlusion for10-min followed by 7-day reperfusion. Effect of monosodium glutamate, ketamine on DNDin the hippocampus was examined by counting neuronal density in CA_1 sector. The Ca~(2+)content in dorsal hippocampus was measured, and the effect of ketamine on Ca~(2+) concen- tration was examined. The results showed that the contents of glutamate and aspartate indorsal hippocampus were increased significantly at the moment of 10-min transient fore-brain ischemia (P0.05). These results suggested that excitatory amino acids (EAA)and Ca~(2+) played an important role directly or indirectly in the development of DND inhippocampus following transient forebrain ischemia in gerbils; ca~(2+) over-load may be ancommon final pathway of neuronal death.

Objective To analyze the clinical manifestations,diagnosis,treatment and prognosis of patients with splenic abscess.Method The clinical data,including baseline clinical data,clinical features,past history,pathogen culture result,treatment and the prognosis were retrospectively analyzed in the patients with the discharge diagnosis splenic abscess from January 1991 to March 2012 in Peking Union Medical College Hospital.Results The media time from onset to Peking Union Medical College Hospital of the 19 patients were 29 days.Among them,9 patients were cured,8 were improved and 2 died.Risk factors,such as tumor burden,diabetes,and using immunosuppressive agents etc,can be found in most patients with splenic abscess.All the 19 patients had splenic image changes and non-specific clinical features.The most common three clinical symptoms were fever(18 cases),chills (12 cases) and shivering (11 cases).The most common three signs were abdominal tenderness (9 cases),left upper quadrant sensitive to percussion (7 cases) and splenomegaly (4 cases).The most common etiological culture results were gram negative bacilli (9 cases),gram positive coccus (8 cases),and fungi (4 cases).Conclusions Clinical features are non-specific in splenic abscess patients.Related exam such as ultrasound should be performed on patients with splenic abscess risk factors to avoid misdiagnosis.Empiric antibiotic administration should begin right after the diagnosis based on the image.Pathogen culture should be timely conducted after pus collection.Individual therapeutical protocol should be chosen according to patient's condition.

AIM To study the chemical constituents from Polygonum hydropiper L..METHODS The petroleum ether,ethyl acetate and n-butanol fractions of 95％ ethanol extract from P.hydropiper were isolated and purified by silica,Sephadex LH-20 and recrystallization,then the structures of obtained compounds were identified by spectral data.RESULTS Thirteen compounds were isolated and identified as β-sitosterol (1),aniba dimer A (2),succinic acid (3),quercetin (4),gallic acid (5),daucosterol (6),quercetin-3-O-β-D-glucoside (7),kaempferol-3-O-β-D-glucoside (8),quercetin-3-O-β-galactoside (9),kaempferol-3-O-β-galactoside (10),stigmast-4-ene-3β,6a-diol (11),fumaric acid (12),ellagic acid (13).CONCLUSION Compounds 11 and 12 are isolated from genus Polygonum for the first time,compounds 2,8,10 and 13 are first obtained from this plant.

Objective To investigate the expression of Toll like receptor 2 (TLR2) in glioma and its relationship with the malignant biological behavior of glioma, so as to provide a new therapeutic target for glioma immunotherapy. Methods The tumor tissues of 90 glioma patients undergoing surgical excision were collected, of which WHO gradingⅠ-Ⅱgrade 39 cases,Ⅲgrade 24 cases,Ⅳgrade 24 cases. In addition, the normal brain tissues of 12 patients undergoing routine intracranial decompression were selected. The human glioma cell lines U87-MG, U251-MG and human astrocyte cell line HA were cultured. The expression levels of TLR2 mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western Blot test. The correlation between TLR2 protein and different clinical pathological parameters was analyzed. Results The expression levels of TLR2 mRNA and protein in the glioma tissuesⅠ-Ⅱgrade,Ⅲgrade andⅣgrade were significantly higher than those in normal brain tissues (0.27 ± 0.09, 0.57 ± 0.12 and 0.96 ± 0.18 vs. 0.11 ± 0.05; 0.31 ± 0.05, 0.44 ± 0.05 and 0.71 ± 0.09 vs. 0.02 ± 0.01), there were statistical differences (P<0.05). In different grades of glioma tissues, there were statistical differences in the expression levels of TLR2 mRNA and protein (F = 205.9 and 194.9, P<0.05). The expression levels of TLR2 mRNA and protein in human malignant glioma cell lines U87-MG and U251-MG were significantly higher than those in human astrocyte cell line HA (1.000 ± 0.100 and 0.356 ± 0.060 vs. 0.245 ± 0.030, 0.720 ± 0.100 and 1.800 ± 0.150 vs. 0.004 ± 0.000), there were statistical differences (P<0.05). The expression of TLR2 protein was independent of age, sex and location(P>0.05), but was related to tumor diameter and WHO grading (P < 0.01). Conclusions TLR2 in different grade glioma tissues and glioma cell lines are expressed, and its expression level is associated with the malignant degree of glioma; TLR2 protein not only can be used as a biomarker of gliomas and prognosis, but also provide a new target for the treatment of glioma.

Objective To investigate the relationship between white matter lesions (WML) within the cholinergic pathway and vascular cognitive impairment.Method Middle-aged and elderly stroke patients underwent brain MRI examination and Montreal Cognitive Assessment (MoCA).Cholinergic Pathways Hyperintensities Scale (CHIPS) scores and the overall WML burden by Schelten on fluidattenuated inversion recovery MRI images were determined and compared with MoCA scores.Spearman partial rank correlation coefficients and standardized regression coefficients were calculated.Results Thirty four patients were included ( mean age ( 62.2 ± 8.8 ) years, 45-82 years).MoCA scores negatively correlated with WML burdens by Schelten scores ( β = - 0.357, P = 0.042) and CHIPS scores ( β =-0.382,P=0.026).CHIPS scores were negatively associated with visuospatial and executive function (r = - 0.290, P = 0.048 ), naming function ( r = - 0.486, P = 0.002 ), attention ( r = - 0.311, P =0.037) and abstraction ( r = - 0.344, P = 0.023).Schelten scores were negatively associated with naming function (r = - 0.492, P = 0.002), attention ( r = - 0.364, P = 0.017) and abstraction ( r = - 0.390,P=0.011).Conclusion WML lesions within the cholinergic pathyway play a possible role in vascular cognitive impairment especially in visuospatial and executive function.

We described an elderly female with type 2 diabetes referred to our hospital with fever,nausea and upper abdominal pain.The patient had got duodenal tumor and received the pancreaticoduodenectomy (PD) 12 years ago.The laboratory examinations revealed white blood cells (WBC) increasing and severe hypocalcemia.Abdominal computed tomography (CT) revealed a huge gas-forming pyogenic liver abscess (PLA) in left lobe of the liver.The patient got cured after correction of calcium metabolism disorders,treatment with antibiotic and receiving percutaneous tube drainage.We concluded that we should remain on high alert of those patients with DM and the history of cancer,when he or she gets fever of unknown origin and abdominal tenderness.PLA should be considered.

AIM:To observe the effect of B-HT933, a selective α2-adrenoceptor agonist, on lipopolysaccha-ride ( LPS )-induced TNF-αproduction in neonatal rat cardiomyocytes and to explore the underlying mechanisms . METHODS:The neonatal rat cardiomyocytes were cultured .The localization of α2A-adrenoceptor in the cardiomyocytes was examined by immunofluorescence staining .The cardiomyocytes were exposed to LPS or/and B-HT933 for different time.The level of TNF-αin the supernatants and the mRNA expression of TNF-αwere detected by ELISA and real-time PCR, respectively.In addition, LPS-associated signal molecules in the cardiomyocytes were also examined by Western blotting.RESULTS: Immunofluorescence staining showed that α2A-adrenoceptors were localized in the cardiomyocytes . LPS stimulated TNF-αproduction in the cardiomyocytes in a dose and time-dependent manner .B-HT933 pretreatment sig-nificantly inhibited the expression of TNF-αat mRNA and protein levels in LPS-treated cardiomyocytes .Furthermore, LPS exposure induced IκBαand p38 phosphorylation in cardiomyocytes and only IκBαphosphorylation was prevented by B-HT933 treatment.CONCLUSION:α2A-adrenoceptors are present in neonatal rat cardiomyocytes and its agonist B -HT933 inhibits LPS-induced TNF-αproduction in cardiomyocytes via suppressing IκBαphosphorylation .

OBJECTIVE: To study the changes in mRNA and long non-coding RNA (lncRNA) expression profiles in a mouse model of bleomycin-induced lung fibrosis and identify lung fibrosis-related mRNA for coding-noncoding coexpression (CNC) bioinformatics analysis of the differential lncRNAs. METHODS: Lung fibrosis was induced by intratracheal injection of bleomycin in 10 C57BL/6 mice and another 10 mice with intratracheal injection of saline served as the control group. Lung tissues were harvested from the mice at 14 days after the injections and lung fibrosis was assessed using Masson and HE staining. LncRNA chip technology was used to screen the differentially expressed mRNAs and lncRNAs in mice with lung fibrosis, and GO and KEGG pathway analyses of the differential mRNAs were performed using NCBI database and UCSC database to identify possible fibrosis-related mRNAs, which were validated by qRT-PCR to construct a coding and non-coding co- expression network with the differential lncRNAs. RESULTS: Compared with the control mice, the mice with intratracheal injection of bleomycin showed obvious lung fibrosis. The results of gene chip analysis showed that 127 mRNAs were upregulated and 184 mRNAs were down-regulated in the model group as compared with the control group. GO and pathway analysis suggested that the differentially expressed genes participated mainly in immune response, cell differentiation, and cytoskeletons; the involved signal pathways were associated mainly with cytokine and cytokine receptor interaction and chemokine signal transduction. Bioinformatics analysis identified a significant coexpression network between the fibrosisrelated mRNA and the differentially expressed lncRNA. CONCLUSIONS In mice with lung fibrosis, the differential expressions of fibrosis-related mRNAs in the lung tissues are closely correlated with the co- expressions of a large number of differential lncRNAs, which points to a new direction for investigation of the pathogenesis of pulmonary fibrosis.
Animals ; Bleomycin/toxicity* ; Gene Expression Profiling ; Gene Regulatory Networks ; Mice ; Mice, Inbred C57BL ; Pulmonary Fibrosis/genetics* ; RNA, Long Noncoding/genetics* ; RNA, Messenger/genetics*

Objective To investigate the protective effect of 5-hydroxy-6,7-dimethoxyflavone(5-HDF),a compound extracted from Elsholtzia blanda Benth.,against lung injury induced by H1N1 influenza virus and explore its possible mechanism of action.Methods 5-HDF was extracted from Elsholtzia blanda Benth.using ethanol reflux extraction and silica gel chromatography and characterized using NMR and MS analyses.In an A549 cell model of H1N1 influenza virus infection(MOI=0.1),the cytotoxicity of 5-HDF was assessed using MTT assay,and its effect on TRAIL and IL-8 expressions was examined using flow cytometry;Western blotting was used to detect the expression levels of inflammatory,apoptosis,and ferroptosis-related proteins.In a mouse model of H1N1 influenza virus infection established by nasal instillation of 50 μL H1N1 virus at the median lethal dose,the effects of 30 and 60 mg/kg 5-HDF by gavage on body weight,lung index,gross lung anatomy and lung histopathology were observed.Results 5-HDF exhibited no significant cytotoxicity in A549 cells within the concentration range of 0-200 μg/mL.In H1N1-infected A549 cells,treatment with 5-HDF effectively inhibited the activation of phospho-p38 MAPK and phospho-NF-κB p65,lowered the expressions of IL-8,enhanced the expression of anti-ferroptosis proteins(SLC7A11 and GPX4),and inhibited the expressions of apoptosis markers PARP and caspase-3 and the apoptotic factor TRAIL.In H1N1-infected mice,treatment with 5-HDF for 7 days significantly suppressed body weight loss and increment of lung index and obviously alleviated lung tissue pathologies.Conclusion 5-HDF offers protection against H1N1 influenza virus infection in mice possibly by suppressing H1N1-induced ferroptosis,inflammatory responses,and apoptosis via upregulating SLC7A11 and GPX4,inhibiting the activation of phospho-NF-κB p65 and phospho-p38 MAPK,and decreasing the expression of cleaved caspase3 and cleaved PARP.

Objective This study aimed to explore the expression trends of innate immune cells and immune-checkpoint molecules validated by data calculation in the process of oral mucosal carcinogenesis,as well as to explore methods of suppressing oral mucosal carcinogenesis based on immunotherapy by predicting their interactions.Me-thods 1)The cancer genome atlas(TCGA)database comprehensively scores immune cells and immune-checkpoint molecules in the process of oral mucosal carcinogenesis and screens out intrinsic immune cells and immune-check-point molecules that interfere with tumor immune escape.2)Clinical patient blood routine data were collected for the statistical analysis of peripheral blood immune cells during the progression of oral mucosal carcinogenesis.Immune cells in peripheral blood that may affect the progression of oral mucosal carcinogenesis were screened.3)Immunohis-tochemical staining was performed on intrinsic immune cells and immune-checkpoint molecules validated based on da-ta calculation in various stages of oral mucosal carcinogenesis.4)Special staining was used to identify innate immune cells in various stages of oral mucosal carcinogenesis based on data-calculation verification.5)Survival analysis was conducted on intrinsic immune cells and immune-checkpoint molecules validated based on data calculation during the process of oral mucosal carcinogenesis.The association of intrinsic immune cells and immune-checkpoint molecules with the prognosis of oral squamous cell carcinoma was verified.Results The expression of monocytes and neutro-phils increased during the process of oral mucosal carcinogenesis.The expression of eosinophils showed a single peak trend of up and down.The expression of mast cells decreased.In the process of oral mucosal carcinogenesis,the ex-pression of the immune-checkpoint molecules cytotoxic T-lymphocyte-associated protein 4(CTLA4)and programmed cell death-ligand(PD-L1)increased.The expression trends of monocytes,neutrophils,and eosinophils were positively correlated with those of CTLA4 and PD-L1 immune-checkpoint molecules.The expression trend of mast cells was negatively correlated with the expression of CTLA4 and PD-L1.Monocytes,neutrophils,and eosinophils may pro-mote tumor immune escape mediated by CTLA4 and/or PD-L1,thereby accelerating the progression of oral mucosal carcinogenesis.Mast cells may inhibit tumor immune escape mediated by CTLA4 and/or PD-L1,delaying the progres-sion of oral mucosal carcinogenesis.Conclusion Therefore,interference with specific immune cells in innate immu-nity can regulate the expression of CTLA4 and/or PD-L1 to a certain extent,inhibit tumor immune escape,and delay the progression of oral mucosal carcinogenesis.