Epilepsy is one of the nervous system diseases,which is correlated with multiple pathogenic factors and caused by repeated discharge of neurons.Currently,there are more than 50 million people worldwide suffering from epilepsy with an average annual increase of 100 000 cases.The prevalence rate of sleep disorders in epilepsy patients is high,by up to two times than that of healthy subjects.Common sleep disorders in epilepsy patients include insomnia,sleep apnea,restless legs syndrome and parasomnias.The characteristics of sleep abnormalities have differences with the epilepsy syndromes.The relationship between epilepsy and sleep is complex and interactive.The mechanism of combined sleep disorders in epilepsy patients is still unclear.In this paper,the relationship between epilepsy and sleep disorders in epilepsy patients was summarized,which involves multiple aspects such as the possible mechanism of combined sleep disorders,the common features of sleep disorders,the possible mechanism of sleep abnormalities in different epilepsy syndromes and the characteristics of sleep structure,the effect of antiepileptic drugs on sleep architecture abnormalities and its role in combined sleep disorders.

Objective To investigate the characteristic of sleep disorders in patients with frontal lobe epilepsy (FLE) through polysomnography and subjective questionnaires. Methods Eighteen FLE patients (FLE group) and 15 healthy subjects (control group) were recruited to our study. Two groups were evaluated by whole-night polysomnography, included total time in bed (TIB), total sleep time (TST), sleep efficiency (SE), sleep latency (SL), rapid eye movement (REM) latency (RL), wake after sleep onset (WASO), the percentage of non-REM (NREM) 1, 2, 3 stages and the percentage of REM occupied TST, the apnea-hypopnea index (AHI), hypopnea index, mean oxygen saturation (SpO2) and nadir oxygen saturation, periodic leg movements (PLMs) index and PLMs index of REM sleep, sleep stage shifts (SSS) and sleep stage shifts per hour (SSS/h), wake, NREM1, NREM2, NREM3 and REM sleep stage shifts (abbreviated as N1, N2, N3, REM, W) and their proportions of SSS (abbreviated as N1/SSS, N2/SSS, N3/SSS, REM/SSS, W/SSS). Data of depress and fatigue degrees, sleep quality, excessive daytime sleepiness were assessed by subjective questionnaires. Results Compared with control group, TIB [(503.22±62.33) min], WASO[(54.11±18.97) min], SSS (161.17±56.83), SSS/h (22.60±6.25), N1(42.56± 15.26) and N2 (57.28 ± 26.57), number of wake stage (25.33 ± 10.97) were significantly increased in FLE group. Meanwhile, scores of depression (9.33±5.01) and fatigue (5.72±3.36) were increased in FLE group compared with those of control group. Conclusion The sleep disorders can be found in FLE, which are related with depression and FLE seizure frequency.

The gene encoding urease subunit A (ureA) of Helicobacter pylori (H. pylori) was cloned from H. pylori isolate by polymerase chain reaction (PCR). Sterile distilled water instead of DNA served as negative control. The nucleotide sequence of the amplified product was determined. Homologous analysis of the ureA against that reported by Clayton CL and the GenBank and SwissProt databases were performed with the BLAST program at the Genome Net through the Internet. 0.8 kb PCR product was amplified from all H. pylori clinical isolators. The nucleotide sequence of the ureA was determined. The nucleotide sequence of the ureA began with ATG as the initiation codon and terminated in TAA as stop codon. The coding regions had a 44% G + C content. The DNA sequence was 98% homologous to that reported by Clayton CL (688 out of 702 residues were identical). The derived amino-acid sequences of the ureA were 99% homologous to that reported by Clayton CL (232 out of 234 residues were identical). The nucleotide sequence and the predicted protein showed significant homology to ureA of H. pylori in the NCBI Entrez database.
Base Sequence ; Cloning, Molecular ; DNA, Bacterial/chemistry ; DNA, Bacterial/genetics ; *Genes, Bacterial ; Genetic Code ; Helicobacter Infections/microbiology ; Helicobacter pylori/enzymology ; Helicobacter pylori/*genetics ; Helicobacter pylori/isolation & purification ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Transcription, Genetic ; Urease/*genetics ; Urease/metabolism

AIM: To investigate the changes of expression of SR-BI in phorbol 12-myristate 13-acetate(PMA) differentiated U937 cells.METHODS: U937 cells were cultured with 100 nmol/L PMA in order to differentiate the cells to macrophages.Immunocytochemical method,Western blotting analysis and reverse transcription polymerase chain reaction(RT-PCR) were used to detect SR-BI protein and mRNA during differentiation.RESULTS: Immunocytochemistry showed that after exposure of U937 cells to PMA for 24,48,72 hours,the values of SR-BI protein expression in U937 cells were 15.94?3.56,27.86?4.39 and 9.08?2.37,with the first two higher than that in undifferentiated cells(7.76?1.74,P0.05) increment in the expression of SR-BI protein compared with U937 monocytes.RT-PCR showed that relative SR-BI mRNA expression in different group was 0.112?0.006,0.235?0.014,0.344?0.140 and 0.138?0.010,respectively.CONCLUSION: SR-BI protein and mRNA were increased after differentiation,reached a peak at 48 hours,and decreased at 72 hours.High expression levels of SR-BI in U937 macrophages following PMA differentiation may be correlated with foam cell formation.

Objective To study histone acetylation patterns in peripheral boiood mononuclear cells (PBMC) from type 2 diabetic patients PBMC form 12type 2 diabetic patients and 12 healthy contrcl subiects were collecyed The global histone H3/H4 acetylation was determined by assay kit The differetial expression of the histone acetyltransferases and histone deacetylases were measured by real time PCR.Results increased H3/H4 acetylation was observed in PBMC of type 2 diabetic patients compared with control subjects (H3:0.134±0.035vs0.181±0.032,p<0.05;H4:0.266±0.070VS0.324±0.062,P<0.01).the mRNA levels of p300,CREBBP,HDAC2,HDAC7,and STRTL in patients PBMC compared with healthy control subieects were 3.11±0.38,2.78±0.45,3.55±0.30,0.77±0.37 and 0.40±0.25 folds respectively conclusion histone acetylation appears abnormal in PBMC of type 2 diabetic patients.

The gene encoding urease subunit A (ureA) of Helicobacter pylori (H. pylori) was cloned from H. pylori isolate by polymerase chain reaction (PCR). Sterile distilled water instead of DNA served as negative control. The nucleotide sequence of the amplified product was determined. Homologous analysis of the ureA against that reported by Clayton CL and the GenBank and SwissProt databases were performed with the BLAST program at the Genome Net through the Internet. 0.8 kb PCR product was amplified from all H. pylori clinical isolators. The nucleotide sequence of the ureA was determined. The nucleotide sequence of the ureA began with ATG as the initiation codon and terminated in TAA as stop codon. The coding regions had a 44% G + C content. The DNA sequence was 98% homologous to that reported by Clayton CL (688 out of 702 residues were identical). The derived amino-acid sequences of the ureA were 99% homologous to that reported by Clayton CL (232 out of 234 residues were identical). The nucleotide sequence and the predicted protein showed significant homology to ureA of H. pylori in the NCBI Entrez database.
Base Sequence ; Cloning, Molecular ; DNA, Bacterial ; chemistry ; genetics ; Genes, Bacterial ; Genetic Code ; Helicobacter Infections ; microbiology ; Helicobacter pylori ; enzymology ; genetics ; isolation & purification ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Transcription, Genetic ; Urease ; genetics ; metabolism

Multidrug resistance (MDR) is one of the main causes leading to the failure in cancer treatment. Differential proteins between esophageal squamous cell carcinoma (ESCC) cell line EC9706 and its cisdiamminedichloroplatinum (CDDP)-resistant subline EC9706/CDDP revealed by quantitative analysis may provide deeper insights into the molecular mechanisms of MDR implicated in ESCC. EC9706/CDDP was generated by exposure of its parental sensitive EC9706 to a step-wise increase of CDDP concentration during EC9706 cultivation. The stable isotope labeling with amino acids in cell culture (SILAC) was used to label EC9706 and EC9706/CDDP with heavy and light medium, separately. Mixed peptides derived from EC9706 and EC9706/CDDP were analyzed by high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS/MS) and subsequently subjected to bioinformatics analysis to identify differential proteins between EC9706 and EC9706/CDDP. Compared to parental EC9706, EC9706/CDDP manifested phenotypes of slow proliferation, cell pleomorphology, atypia and increased resistant-index 3.23. Seventy-four differential proteins identified in the present study belongs to various families with multiple functions, such as cytoskeleton (20%), energy metabolism (11%), transcription regulation and DNA repair (11%), redox homeostasis (9.5%), protein biosynthesis and mRNA processing (12%), ribosome constituent (8.1%), molecular chaperone (8.1%), immunity/inflammation (5.4%), intracellular transport (5.4%) and nucleosome assembly (2.7%), which indicated that development of MDR is a complicated process involving dysregulation of multiple molecules and pathways. The data is of great value for in-depth elucidation of molecular mechanisms of the MDR implicated in ESCC and may represent potential molecular targets for future therapeutic development.

BACKGROUND:Because of its anti-inflammatory and antioxidant activities,Apelin-13 plays an effective role in the treatment of common clinical diseases such as neuroinflammation,cardiovascular injury and pneumonia.However,there is no relevant basic research on whether Apelin-13 also has a good effect in the treatment of inflammatory bone loss. OBJECTIVE:To explore the therapeutic effect and mechanism of Apelin-13 on inflammatory bone loss,in order to find potential drugs for the treatment of inflammatory bone loss. METHODS:(1)In vitro experiment:RAW264.7 cells were divided into three groups:control group,lipopolysaccharide group and treatment group.The control group was only added with DMEM complete medium;lipopolysaccharide group was added with lipopolysaccharide(100 ng/mL)induced inflammation DMEM medium;and the treatment group was added with 10 nmol/L Apelin-13+lipopolysaccharide induced inflammation DMEM medium.Then,24 hours after lipopolysaccharide induced inflammation,western blot was used to detect the marker proteins inducible nitric oxide synthase and CD86 of M1 macrophages,and cell immunofluorescence was extracted to detect the expression of inducible nitric oxide synthase.Finally,the same amount of receptor activator of nuclear factor-κB ligand(RANKL;50 ng/ml)was added to the control group,lipopolysaccharide group and treatment group to induce osteoclasts.The results of osteoclast induction were evaluated by tartrate-resistant acid phosphatase staining and F-actin staining after 6 days of induction.(2)In vivo experiment:Eighteen male C57bl/6 mice were randomly divided into three groups:sham group,lipopolysaccharide group and treatment group.The sham group received intraperitoneal injection of 0.1 mL of PBS;the lipopolysaccharide group was injected with 0.1 mL of PBS diluent containing lipopolysaccharide(5 mg/kg);and the treatment group was injected with 0.1 mL of PBS diluent containing lipopolysaccharide(5 mg/kg)+Apelin-13(100 μg/kg).After 7 days of continuous intraperitoneal injection,the mice in each group were killed on the 8th day,and two femurs of each mouse were collected.Half of them were scanned by micro-CT and analyzed by bone mineral density,and the other half were stained by hematoxylin-eosin staining RESULTS AND CONCLUSION:(1)In vitro experiment:Western blot results showed that the expressions of inducible nitric oxide synthase and CD86 in the lipopolysaccharide group were significantly higher than those in the control group,and Apelin-13 could significantly inhibit the M1 polarization of macrophages induced by lipopolysaccharide.Cell immunofluorescence results also showed that the expression of inducible nitric oxide synthase in the treatment group was lower than that in the lipopolysaccharide group.Besides,tartrate-resistant acid phosphatase staining and F-actin staining results showed that Apelin-13 inhibited the abnormal activation and bone resorption of lipopolysaccharide induced osteoclasts.(2)In vivo experiment:The results of micro-CT showed that systemic inflammation led to significant bone loss in the distal femur,while Apelin-13 could significantly inhibit bone loss in vivo.Hematoxylin-eosin staining results also showed that Apelin-13 could effectively alleviate inflammation induced bone loss in the distal femur of mice.To conclude,Apelin-13 can alleviate bone loss induced by systemic inflammation by inhibiting M1 polarization of macrophages,inhibiting abnormal activation of osteoclasts and bone resorption.

ObjectiveTo investigate the mechanism by which modified Shengjiangsan （MSJS） improves diabetic kidney disease （DKD） by activating mitochondrial autophagy. MethodsSixty SPF-grade male Sprague-Dawley rats aged 7-8 weeks were selected. A DKD model was established using a high-sugar， high-fat diet combined with intraperitoneal injection of streptozotocin （STZ）. After successful modeling， the rats were randomly divided into six groups： a normal control group， a model group， low-， medium-， and high-dose MSJS groups （7.7， 15.4， 30.8 g·kg^-1， respectively）， and an irbesartan group （0.384 g·kg^-1）. Each group received either normal saline or the corresponding drug by gavage once daily for 28 consecutive days. Blood glucose， body weight， and kidney weight were recorded. Serum creatinine （SCr） and blood urea nitrogen （BUN） levels were detected using an automatic blood analyzer. Enzyme-linked immunosorbent assay （ELISA） was used to determine urinary microalbumin （mALB）， and serum levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6）. Histopathological changes in renal tissues were observed using hematoxylin-eosin （HE） staining， periodic acid-Schiff （PAS） staining， and transmission electron microscopy （TEM）. The expression levels of mitochondrial autophagy-related proteins in renal tissues were analyzed by Western blot. Immunofluorescence co-localization was employed to detect the co-expression of microtubule-associated protein 1 light chain 3 beta （LC3B） and cytochrome c oxidase subunit Ⅳ （COX Ⅳ）. ResultsCompared with the normal control group， the model group exhibited significant increases in renal index， blood glucose， and 24-hour urinary microalbumin （24 h mALB） （P<0.05， P<0.01）. The levels of serum SCr and BUN were significantly elevated （P<0.01）， and the serum levels of TNF-α， IL-1β， and IL-6 were markedly upregulated （P<0.01）. Histopathological examination revealed glomerular hypertrophy， mesangial expansion and increased deposition， podocyte foot process flattening and fusion， a decreased number of autophagosomes accompanied by mitochondrial swelling， vacuolar degeneration of renal tubular epithelial cells， and inflammatory cell infiltration in the renal interstitium. The expression levels of autophagy-related proteins LC3B， PTEN-induced putative kinase 1 （PINK1）， and E3 ubiquitin-protein ligase （Parkin） were significantly decreased （P<0.05， P<0.01）， while expression of the selective autophagy adaptor protein p62 was significantly increased （P<0.01）. Immunofluorescence signal intensity and LC3B-COX Ⅳ co-expression were both diminished. Compared with the model group， the MSJS treatment groups and the irbesartan group showed significant reductions in renal index， blood glucose， and 24 h mALB （P<0.05， P<0.01）. The serum SCr and BUN levels decreased significantly （P<0.05） and TNF-α， IL-1β， and IL-6 levels were significantly downregulated （P<0.05， P<0.01）. Histopathological damage was alleviated， including reduced glomerular hypertrophy， decreased mesangial deposition， and attenuated podocyte foot process fusion. The number of autophagosomes increased， and mitochondrial swelling was improved. The expression levels of LC3B， PINK1， and Parkin in renal tissues were significantly upregulated， whereas p62 expression was significantly downregulated （P<0.05， P<0.01） in MSJS groups. Immunofluorescence signal intensity was enhanced， and LC3B-COX Ⅳ co-expression was increased. ConclusionMSJS alleviates the inflammatory response in DKD rats and exerts renal protective effects by regulating the PINK1/Parkin signaling pathway and activating mitochondrial autophagy.

ObjectiveTo investigate the mechanism by which modified Shengjiangsan （MSJS） improves diabetic kidney disease （DKD） by activating mitochondrial autophagy. MethodsSixty SPF-grade male Sprague-Dawley rats aged 7-8 weeks were selected. A DKD model was established using a high-sugar， high-fat diet combined with intraperitoneal injection of streptozotocin （STZ）. After successful modeling， the rats were randomly divided into six groups： a normal control group， a model group， low-， medium-， and high-dose MSJS groups （7.7， 15.4， 30.8 g·kg^-1， respectively）， and an irbesartan group （0.384 g·kg^-1）. Each group received either normal saline or the corresponding drug by gavage once daily for 28 consecutive days. Blood glucose， body weight， and kidney weight were recorded. Serum creatinine （SCr） and blood urea nitrogen （BUN） levels were detected using an automatic blood analyzer. Enzyme-linked immunosorbent assay （ELISA） was used to determine urinary microalbumin （mALB）， and serum levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6）. Histopathological changes in renal tissues were observed using hematoxylin-eosin （HE） staining， periodic acid-Schiff （PAS） staining， and transmission electron microscopy （TEM）. The expression levels of mitochondrial autophagy-related proteins in renal tissues were analyzed by Western blot. Immunofluorescence co-localization was employed to detect the co-expression of microtubule-associated protein 1 light chain 3 beta （LC3B） and cytochrome c oxidase subunit Ⅳ （COX Ⅳ）. ResultsCompared with the normal control group， the model group exhibited significant increases in renal index， blood glucose， and 24-hour urinary microalbumin （24 h mALB） （P<0.05， P<0.01）. The levels of serum SCr and BUN were significantly elevated （P<0.01）， and the serum levels of TNF-α， IL-1β， and IL-6 were markedly upregulated （P<0.01）. Histopathological examination revealed glomerular hypertrophy， mesangial expansion and increased deposition， podocyte foot process flattening and fusion， a decreased number of autophagosomes accompanied by mitochondrial swelling， vacuolar degeneration of renal tubular epithelial cells， and inflammatory cell infiltration in the renal interstitium. The expression levels of autophagy-related proteins LC3B， PTEN-induced putative kinase 1 （PINK1）， and E3 ubiquitin-protein ligase （Parkin） were significantly decreased （P<0.05， P<0.01）， while expression of the selective autophagy adaptor protein p62 was significantly increased （P<0.01）. Immunofluorescence signal intensity and LC3B-COX Ⅳ co-expression were both diminished. Compared with the model group， the MSJS treatment groups and the irbesartan group showed significant reductions in renal index， blood glucose， and 24 h mALB （P<0.05， P<0.01）. The serum SCr and BUN levels decreased significantly （P<0.05） and TNF-α， IL-1β， and IL-6 levels were significantly downregulated （P<0.05， P<0.01）. Histopathological damage was alleviated， including reduced glomerular hypertrophy， decreased mesangial deposition， and attenuated podocyte foot process fusion. The number of autophagosomes increased， and mitochondrial swelling was improved. The expression levels of LC3B， PINK1， and Parkin in renal tissues were significantly upregulated， whereas p62 expression was significantly downregulated （P<0.05， P<0.01） in MSJS groups. Immunofluorescence signal intensity was enhanced， and LC3B-COX Ⅳ co-expression was increased. ConclusionMSJS alleviates the inflammatory response in DKD rats and exerts renal protective effects by regulating the PINK1/Parkin signaling pathway and activating mitochondrial autophagy.