TGM6 might not be a specific causative gene for spinocerebellar ataxia resulting from genetic analysis and functional study.

OBJECTIVE: To investigate whether TGM6 is a specific causative gene for spinocerebellar ataxia type 35 (SCA35). MATERIALS AND METHODS: The next-generation sequencing (NGS) data consisted of 47 SCA, 762 non-SCA patients and 2827 normal controls were analyzed. The allele frequencies of low frequent and deleterious TGM6 variants were compared. Functional studies were performed in five widely distributed variants (V314M, R342Q, P347L, V391M, L517W). RESULTS: Two TGM6 detrimental variants were identified in one SCA patient, 14 in non-SCA patients and 43 in normal controls, the allele frequencies of TGM6 variants did not differ among the SCA and other controls. Seven reported pathogenic variants (c.7 + 1G > T, c.331C > T, c.1171G > A, c.1478C > T, c.1528G > C, c.1550 T > G and c.1722_1724delAGA) were identified in patients with various neurologic diseases or normal controls. All the 5 widely distributed variants led to destabilization and significantly reduction of enzymatic activity of TG6 as the reported pathogenic mutations. CONCLUSIONS: TGM6 might not be a specific causative gene for SCA35, the relevant clinical consult or diagnostic should be pay more attention.

Predictive factors for early hypothyroidism following the radioactive iodine therapy in Graves' disease patients.

BACKGROUND: Radioactive iodine (RAI) therapy is an important treatment option for Graves' disease (GD), the main side effect of RAI treatment is hypothyroidism, and the factors resulting in hypothyroidism are still controversial. The purpose of this retrospective study was to clarify the possible risk factors of early hypothyroidism after RAI therapy in Graves' disease. METHODS: We reviewed 312 GD patients treated with RAI between January 2017 to December 2018, collected the potential risk factors, and analyzed the relationship between these variables and early hypothyroidism. RESULTS: After 6 months' follow-up, 218 (69.87%) patients were evaluated as early hypothyroid. Male gender, shorter duration of disease, smaller thyroid weight, lower 2-h radioactive iodine uptake (RAIU), 6-h RAIU, 24-h RAIU and 6/24-h uptake ratio, lower administered dosages were significantly associated with early hypothyroidism. Logistics regression analysis showed that male gender, smaller thyroid weight and lower 6-h RAIU were associated with early hypothyroidism. Multi-factors combined ROC curve analysis suggested that the predictive power of male gender, smaller thyroid weight and lower 6-h RAIU for early hypothyroidism was 0.711. CONCLUSIONS: Our results show that RAI is an effective therapy for GD and most of the cured patients became to hypothyroid within 6 months. Male gender, smaller thyroid weight and lower 6-h RAIU are the main risk factors for early hypothyroidism.

Allicin alleviates inflammation of diabetic macroangiopathy via the Nrf2 and NF-kB pathway.

As diabetic macroangiopathy is becoming increasingly prevalent, it is urgent to explore preventive and therapeutic drugs and study the mechanism. Diabetic mice were induced by intraperitoneal injection of streptozotocin (STZ)for five consecutive days. Diabetic mice were divided into diabetic and allicin groups. After sacrifice, frozen aortic root sections were immunohistochemically stained for nuclear factor erythroid 2-related factor 2 (Nrf2) and inflammation cytokine-tumor necrosis factor α (TNF-α), and the remaining aortic tissues were analyzed by Western blot for the expression of proinflammation genes. In vitro, Nrf2 and inflammatory relative protein expression levels in Human Umbilical Vein Endothelial Cells (HUVECs) were examined. HUVECs proliferation and apoptosis were measured. TNF-α expression was increased in diabetic group compared to that in control group; this effect was alleviated in allicin-treated mice. Inflammation relative protein expression of Vascular Cell Adhesion Molecule 1(VCAM-1), Matrix metalloproteinase 2 (MMP-2), Inducible Nitric Oxide Synthase (iNOS), and monocyte chemotactic protein 1 (MCP-1) was higher in the diabetic group than in the control group; however, allicin treatment inhibited these diabetes-induced increase. In vitro, allicin treatment reversed the hyperglycemia-induced reduction in proliferation, and decreased the apoptosis induced by high glucose. Inflammation relative protein expression was consistent with that in vivo. Additionally, the expression of nuclear factor kappa-B (NF-κB)and Nrf2 was increased in both DM mice and HUVECs; allicin treatment induced a significant reduction in NF-κB level and improvement in Nrf2 level. Allicin alleviates inflammation caused by diabetic macroangiopathy, and the mechanism may occur via increasing Nrf2 and decreasing NF-κB.

Salvianolic acid B improves myocardial function in diabetic cardiomyopathy by suppressing IGFBP3.

BACKGROUND: Salvianolic acid B (Sal B) is the representative component of phenolic acids derived from the dried root and rhizome of Salvia miltiorrhiza Bge. (Labiatae), which has been widely used for the treatment of cardiovascular and cerebrovascular diseases. However, the effect of Sal B on diabetic cardiomyopathy (DCM) is still unclear. METHODS: Type 1 diabetes mellitus was induced in C57BL/6 J mice by streptozotocin (STZ) treatment, whereas meanwhile Salvianolic Acid B (Sal B (15 or 30 mg/kg/d) was intraperitoneally injected for 16 weeks. At the end of this period, cardiac function was assessed by echocardiography, and total collagen deposition was evaluated by Masson's trichrome and Picrosirius Red staining. Human umbilical vein endothelial cells exposed to hypoxia were used to investigate the effect of different doses of Sal B on angiogenesis and tube formation in vitro. Transcriptome sequencing was performed to identify potential targets of Sal B. RESULTS: Sal B ameliorated left ventricular dysfunction and remodeling, and decreased collagen deposition in the heart of diabetic mice. Administration of Sal B increased the expression of vascular endothelial growth factor (VEGF) receptor 2 (VEGFR2) and VEGFA in a dose-dependent manner and promoted angiogenesis both in vivo and in vitro. Furthermore, Sal B reduced HG-induced insulin-like growth factor-binding protein 3 (IGFBP3) expression, induced the phosphorylation of extracellular signal-regulated protein kinase and protein kinase B (AKT) activities, enhanced cell proliferation, and activated VEGFR2/VEGFA signaling in endothelial cells. The underlying mechanisms involve SalB that enhances IGFBP3 promoter DNA methylation and induce nuclear translocation of IGFBP3 in HUVECs under hypoxia. CONCLUSIONS: Sal B promoted angiogenesis and alleviated cardiac fibrosis and cardiac remodeling in DCM by suppressing IGFBP3.

Mutation screening in Chinese patients with familial Alzheimer's disease by whole-exome sequencing.

Familial Alzheimer's disease (FAD) is characterized by a positive family history of dementia and typically occurs at an early age with an autosomal dominant pattern of inheritance. Amyloid precursor protein (APP), presenilin1 (PSEN1), and presenilin2 (PSEN2) are the major causative genes of FAD. The spectrum of mutations in patients with FAD has been investigated extensively in the Caucasian population but rarely in the Chinese population. Here, we performed whole-exome sequencing in a total of 15 unrelated Chinese patients with FAD. Among them, 12 were found to carry missense variants in APP, PSEN1, and PSEN2. Two novel variants (APP: p.D244G, p.K687Q), 3 variants not previously associated with FAD (APP: p.T297M, p.D332G; PSEN1: p.R157S), and 7 previously reported pathogenic variants (APP: p.V717I; PSEN1: p.M139I, p.T147I, p.L173W, p.F177S, p.R269H; PSEN2: p.V139M) were identified. The novel variant APP p.K687Q was classified as likely pathogenic, and the other 4 variants (APP: p.D244G, p.T297M, p.D332G; PSEN1: p.R157S) were classified as uncertain significance. Therefore, APP, PSEN1, and PSEN2 mutations account for 2 (25.0%), 5 (62.5%), and 1 (12.5%) of the genotyped cases positive for mutations, respectively. Furthermore, the genotype-phenotype correlations were described. Our findings broaden the genetic spectrum of FAD with APP, PSEN1, and PSEN2 variants.

Expression of long non­coding RNA and mRNA in the hippocampus of mice with type 2 diabetes.

Long non­coding RNAs (lncRNAs) serve key roles in cell growth, development and various diseases associated with the central nervous system. However, differential expression profiles of lncRNAs in type 2 diabetes have not been reported. The present study aimed to analyze the expression pattern of lncRNA­mRNA in a type 2 diabetic mouse model using microarray analysis. The mouse model of type 2 diabetes was established and the total RNAs were extracted from the hippocampus of the mice used in the present study. The total RNAs were then examined by the GeeDom human lncRNA + mRNA V4.0 expression profile and analyzed through comparing Gene Ontology (GO) enrichment analysis and signal pathway analysis with the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. There were statistically significant differences between the expression of IncRNAs and mRNA in the healthy mice and that of the diabetic mice. In the diabetic mice, 130 different lncRNAs were expressed with 126  significantly upregulated and 4 significantly downregulated and 49 different mRNAs were detected with 45 significantly upregulated and 4 downregulated. GO analysis indicated that the mRNAs that are affected are involved in transport, cell adhesion, ion transport and metabolic processes. KEGG and Reactome enrichment analysis indicated that mRNAs impact on cholinergic synapses, nuclear factor­kB pathway, Toll like receptor 4 cascade and zinc transporter are correlated with cognitive dysfunction in type 2 diabetes. A dynamic lncRNA­mRNA network was constructed containing 123 lncRNAs and 48 mRNAs, which can elucidate the interaction between lncRNA and mRNA. Overall, this is the first study to indicate that lncRNAs are differentially expressed in the type 2 diabetic mice.

Association between serum uric acid level and multiple system atrophy: A meta-analysis.

OBJECTIVES: Lower serum uric acid (UA) levels are considered to be related to the risk to develop many neurodegenerative disorders. However, the association between serum UA level and multiple system atrophy (MSA) remains controversial. The aim of this meta-analysis was to evaluate the relationship between serum UA level and MSA. PATIENTS AND METHODS: PubMed, Web of Science, Embase, Cochrane Library and China National Knowledge Infrastructure (CNKI) were searched for eligible studies. Standardized mean difference (SMD) and 95% confidence intervals (95% CI) were calculated in a fixed-effects model or a random-effects model when appropriate. Subgroup analyses were carried out based on gender. A total of 6 eligible studies involving 547 MSA patients and 637 healthy individuals were identified. RESULTS: Meta-analysis results revealed that individuals with MSA had lower sera levels of UA as compared with healthy controls (pooled SMD is -0.51, 95%CI: -0.88 to -0.14; p = 0.006). The subgroup analysis to detect sex differences showed that the pooled SMD was -0.61 (95% CI: -0.82 to -0.40; p < 0.0001) for males and -0.22 (95% CI: -0.55 to 0.10; p = 0.18) for females compared with healthy controls. CONCLUSION: Our meta-analysis revealed that lower serum level of UA is associated with an increased risk of MSA and the relationship is significant in men but not in women.

Paeonol exerts potential activities to inhibit the growth, migration and invasion of human gastric cancer BGC823 cells via downregulating MMP­2 and MMP­9.

Paeonol (Pae) is an herbal extract that has attracted extensive attention for its anti­cancer effects demonstrated by a number of studies, which have predominantly demonstrated inhibition of cell proliferation and induction of apoptosis. The influence of Pae on cancer cell metastasis has been less widely reported. The present study aimed to investigate the under­reported effects of Pae on the growth, invasion and migration of poorly differentiated BGC823 gastric cancer cells with strong invasive and metastatic abilities. The anti­proliferative and pro­apoptotic effects of Pae on BGC823 cells were verified by Cell Counting kit­8 and Annexin V­fluorescein isothiocyanate/propidium iodide assays. Cell scratch­wound healing and Transwell methods were applied, and it was demonstrated that Pae could exert inhibitory activities on the invasion and migration of BGC823 cells. Furthermore, it was indicated by western blot analysis that Pae could downregulate the protein expression levels of matrix metalloproteinase (MMP)­2 and ­9 in a concentration­dependent manner, which may support a novel potential mechanism accounting for its anti­cancer effects on gastric cancer.

Allicin improves carotid artery intima-media thickness in coronary artery disease patients with hyperhomocysteinemia.

Homocysteine (Hcy) is an important and independent risk factor for atherosclerotic diseases, such as coronary artery disease and ischemic cerebrovascular disease. Increased carotid artery intima-media thickness (IMT) is a non-invasive marker of systemic atherosclerosis. Allicin treatment may decrease serum Hcy levels and improve impaired endothelial function in rats with hyperhomocysteinemia (HHcy). The present study hypothesized that allicin has an anti-atherosclerotic effect in coronary heart disease and tested the effects of allicin treatment on carotid artery IMT and plasma Hcy levels in coronary heart disease patients with HHcy. Sixty-two coronary heart disease patients with HHcy were randomly divided into an allicin group and a control group. All patients underwent diagnostic assessment, plasma Hcy assay, blood lipid measurement and B-mode ultrasound of the carotid artery prior to and after treatment. Plasma Hcy levels were determined by high-performance liquid chromatography and fluorescence detection. Carotid artery IMT was calculated using an automated algorithm based on a validated edge-detection technique. After 12 weeks, significant decreases in carotid artery IMT, plasma Hcy levels, total cholesterol and triglycerides were observed in the allicin group (all P<0.05), and the decreases in the allicin group were significantly greater than those in the control group (all P<0.01). These findings suggested that reducing plasma Hcy levels may be useful for preventing the generation and development of atherosclerosis in patients with coronary heart disease. Allicin was able to decrease Hcy levels, total cholesterol and triglycerides as well as carotid artery IMT.

Decreased gene expression of CD2AP in Chinese patients with sporadic Alzheimer's disease.

Many sporadic Alzheimer's disease (SAD) risk genes have been identified in the last decades, but most of them have not been consistently accepted. Here, we sought to identify SAD-associated genes and their potential mechanisms involved in SAD pathogenesis. A 2-stage design was employed. In stage 1, 95 variants in 75 genes that were previously reported as SAD-risk genes in Caucasian populations were evaluated in 1857 subjects (422 SAD patients and 1435 controls). In stage 2, a subset of promising variants found in stage 1 were further evaluated in an independent cohort of 1001 subjects (254 SAD and 747 controls). Variants in CD2AP were significantly associated with SAD risk in our subjects. Furthermore, CD2AP gene expression in peripheral blood lymphocytes (PBL) from 209 SAD patients and 213 controls was determined. CD2AP gene expression in PBL was significantly decreased in patients with SAD as compared with controls. Our study suggests that CD2AP is an SAD-risk gene in Chinese Han population and CD2AP gene expression is decreased in the PBL of patients with SAD, indicating its possible systemic involvement in SAD.

Protective effect of allicin on high glucose/hypoxia-induced aortic endothelial cells via reduction of oxidative stress.

This study was designed to explore the protective effect of allicin on aortic endothelial cell injury induced by high glucose/hypoxia and to investigate the corresponding mechanisms. The primary-cultured murine aortic endothelial cells were subcultured. The third passage of cells was adopted and randomly divided into five groups: The normal group (NG), the mannitol group (MG), the high-glucose/hypoxia group (HG), the allicin group (AG) and the protein kinase C (PKC) inhibitor group (GG). The general morphology was observed under an inverted phase-contrast microscope and cell viability was assessed using the MTT assay. Intracellular reactive oxygen species (ROS) levels in the endothelial cells were quantified using dihydroethidium staining. The levels of 8-hydroxydeoxyguanosine (8-OHdG), nuclear factor-κB (NF-κB), NADPH oxidase 4 (Nox4) and hypoxia-inducible factor-1α (HIF-1α) and the activity of PKC were measured using ELISA. A quantitative polymerase chain reaction (qPCR) was adopted to evaluate the mRNA expression of Nox4, HIF-1α and NF-κB. The altered cell morphology observed in HG was notably ameliorated in the AG and GG. The protein levels of 8-OHdG, NF-κB, Nox4, HIF-1α and PKC in the HG were higher than those in the other groups. Furthermore, the cell viability in the AG was significantly increased and the protein levels of 8-OHdG, NF-κB, Nox4, HIF-1α and PKC were significantly decreased compared with those in the HG. The ROS production was found to be increased in the HG cells, while there was a significant decrease in the AG cells. These data indicate that allicin exerts a protective effect against high glucose/hypoxia-induced injury in aortic endothelial cells through its antioxidative action, which may involve the inhibition of the PKC pathway and regulation of HIF-1α.

APOE4 enhances age-dependent decline in cognitive function by down-regulating an NMDA receptor pathway in EFAD-Tg mice.

BACKGROUND: Alzheimer's disease (AD) causes progressive loss of memory and cognition, exacerbated by APOE4, the greatest genetic risk factor for AD. One proposed mechanism for apolipoprotein E (apoE) effects on cognition is via NMDAR-dependent signaling. APOE genotype-specific effects on this pathway were dissected using EFAD-transgenic (Tg) mice (5xFAD mice, that over-express human amyloid-beta (Aß) via 5 familial-AD (FAD) mutations, and express human apoE), and 5xFAD/APOE-knockout (KO) mice. Previous data from EFAD-Tg mice demonstrate age-dependent (2-6 months), apoE-specific effects on the development of Aß pathology. This study tests the hypothesis that apoE4 impairs cognition via modulation of NMDAR-dependent signaling, specifically via a loss of function by comparison of E4FAD mice with 5xFAD/APOE-KO mice, E3FAD and E2FAD mice. RESULTS: Using female E2FAD, E3FAD, E4FAD and 5xFAD/APOE-KO mice aged 2-, 4-, and 6-months, the Y-maze and Morris water maze behavioral tests were combined with synaptic protein levels as markers of synaptic viability. The results demonstrate a greater age-induced deficit in cognition and reduction in PSD95, drebrin and NMDAR subunits in the E4FAD and 5xFAD/APOE-KO mice compared with E2FAD and E3FAD mice, consistent with an apoE4 loss of function. Interestingly, for NMDAR-mediated signaling, the levels of p-CaMK-II followed this same apoE-specific pattern as cognition, while the levels of p-CREB and BDNF demonstrate an apoE4 toxic gain of function: E2FAD > E3FAD > 5xFAD/APOE-KO > E4FAD. CONCLUSION: These findings suggest that compared with E2FAD and E3FAD, E4FAD and 5xFAD/APOE-KO mice exhibit enhanced age-induced reductions in cognition and key synaptic proteins via down-regulation of an NMDAR signaling pathway, consistent with an apoE4 loss of function. However, levels of p-CREB and BDNF, signaling factors common to multiple pathways, suggest a gain of toxic function. Publications in this field present contradictory results as to whether APOE4 imparts a loss or gain of function. As with the results reported herein, the overall effect of APOE4 on a given CNS-specific measure will be the product of multiple overlapping mechanisms. Thus, caution remains critical in determining whether APOE gene inactivation or therapies that correct the loss of positive function related to apoE4, are the appropriate therapeutic response.

Experiences of patients with abnormal extubation of PICC tubes: a qualitative study.

OBJECTIVE: To explore the experiences of patients with abnormal extubation of PICC tubes. METHODS: Using phenomenological research methods, 15 cases of patients with abnormal extubation of PICC tubes were enrolled in semi-structured interviews. Data were analyzed by Nancy's phenomenological procedure. RESULTS: After abnormal extubation, patients exhibited conflicting complicated mood which combined negative experience and positive experience. Negative experience was mainly for complaint, helpless, worry and fear. Positive experience was mainly for relief and peace of mind. CONCLUSIONS: Patients with abnormal extubation often possessed negative experience. So nursing staff should be suggested to communicate with patients before extubation in order to reduce the dispute between nurses and patients. At the same time, we should summarize and analyze the reasons and factors for abnormal extubation, and take targeted intervention measures in clinical to ensure the safety and effectiveness of PICC extubation.

Progranulin-derived Atsttrin directly binds to TNFRSF25 (DR3) and inhibits TNF-like ligand 1A (TL1A) activity.

Atsttrin, a progranulin (PGRN)-derived molecule composed of three TNFR-binding domains of PGRN, binds to TNF receptors (TNFR) and is therapeutic against inflammatory arthritis. Here we screened the associations of Atsttrin and other members in TNFR subfamily, which led to the discovery of TNFRSF25 (DR3) as an additional Atsttrin-interacting member in TNFR family. Similar to TNFR1 and TNFR2, DR3 also directly bound to Atsttrin. The first three cysteine-rich domains (CRD) in the extracellular portion of DR3 were required for this interaction. Atsttrin inhibited the interaction between DR3 and its TNF-Like Ligand 1A (TL1A). In addition, Atsttrin inhibited TL1A-stimulated target gene expressions and neutralized TL1A-enhanced osteoclastogenesis in vitro. Furthermore, Atsttrin ameliorated the pathology in dextran sulfate sodium induced colitis. Taken together, these findings not only provide the new insights into Atsttrin's therapeutic action in inflammatory arthritis, but may also present Atsttrin as a novel biological agent for treating various types of diseases associated with TL1A/DR3 pathway.

Effects of allicin on hyperhomocysteinemia-induced experimental vascular endothelial dysfunction.

This study was designed to investigate the effect and mechanism of allicin on hyperhomocysteinemia-induced experimental vascular endothelial dysfunction in rats. Fifty male Wistar rats were randomly divided into five groups: the normal control rats (NC), the high-methionine-diet rats (Met), the high-methionine-diet rats treated with folic acid, vitaminB6 and vitaminB12 (Met+F), or with low-dose allicin (Met+L), or with high-dose allicin (Met+H). After 6 weeks, we collected blood samples of all groups to determine plasma endothelin (ET), serum homocysteine (Hcy), nitric oxide (NO), superoxide dismutase (SOD), malondialdehyde (MDA), and detected the expression of basic fibroblast growth factors (bFGF), transforming growth factor beta (TGF-ß), tumor necrosis factor-alpha (TNF-α), and intercellular adhesion molecule-1 (ICAM-1) in the aorta. The Hcy and the expression of TGF-ß in both the Met+L and Met+H groups were significantly lower than the Met and Met+F groups. The ET, ET/NO ratio and the MDA levels of the Met+L and Met+H groups were significantly lower than the Met group. The SOD and NO levels and the expression of bFGF, TNF-α and ICAM-1 of the Met+L and Met+H groups were significantly higher than the Met group. Our data indicate that allicin inhibits lipid peroxidation induced by hyperhomocysteinemia and regulates the excretion and equilibrium of ET and NO, and suggest that allicin might be useful in the prevention of endothelial dysfunction caused by hyperhomocysteinemia.

Neuroprotective effects of the Chinese Yi-Qi-Bu-Shen recipe extract on injury of rat hippocampal neurons induced by hypoxia/reoxygenation.

OBJECTIVE: To explore the protective effects of the Chinese Yi-Qi-Bu-Shen recipe (YB) against neuronal injury induced by hypoxia-reoxygenation, which has shown beneficial effect in improving the brain function of type 2 diabetics likely through its antihyperglycemic, antioxidant activity, and investigate its mechanisms. METHODS: The bilateral hippocampus was collected from newborn rats to establish single cell suspension. On the 10th day, the primarily cultured hippocampal neurons were randomly divided into five groups: the normal group (NG), the hypoxia/reoxygenation group (HG), and groups protected with small, medium and large dosages of YB (SG, MG and LG, respectively). The YB-protected groups were treated with different concentrations of YB containing serum before reoxygenation. The metabolic rate of MTT, the malondialdehyde (MDA) content, and the activity of superoxide dismutase (SOD) and lactate dehydrogenase (LDH) were measured with assay kits. The apoptosis rate of hippocampal neurons were tested using flow cytometry analysis. RT-PCR was used to evaluate the mRNA expressions of bcl-2 and bax genes. RESULTS: The SOD activity, the cell survival rate, the bcl-2/bax ratio, and the bcl-2mRNA expression in the HG group were significantly lower (all P<0.01), but the levels of MDA and LDH, the apoptosis rate, and the bax mRNA expression were higher (all P<0.01) than those in the NG group. The SOD activity, the cell survival, the bcl-2mRNA expression, and the bcl-2/bax ratio were significantly higher in all of the YB-protected groups (all P<0.01), but the level of MDA and LDH, the apoptosis rate, and the bax mRNA expression were lower (P<0.01, P<0.05) than those in the HG group in a dose dependent manner. CONCLUSION: The YB extract has a protective effect on hippocampal neurons against injury induced by hypoxia/reoxygenation, through its antioxidant activity and the regulation of apoptosis.

[Outgrowth of neuronal axons on adipose-derived stem cell transplanting for treatment of cerebral infarction in rats].

AIM: To investigate the effects of adipose-derived stem cell (ADSC) transplanting on the outgrowth of neuronal axons and the expressions of GFAP, Neuritin, NF-200 in the brain post focal cerebral ischemia in rats. METHODS: 54 male adult Sprague-Dawley rats were randomly divided into 3 groups: sham-operated group, middle cerebral artery occlusion (MCAO) group and MCAO+ADSC-treated group (n=18 in each group). A permenant focal cerebal ischemia model was established using modified Longa's method ADSC was labeled by DAPI before the transplantation. One day after MCAO, 30 µL of cell suspension containing 1×10(6); cells were injected into the lateral ventricle of MCAO+ADSC-treated group. At 7 d, 14 d and 28 d after MCAO, the expressions of GFAP, Neuritin and NF-200 were detected in ischemic region by Western blot and Immunofluorescence analysis. RESULTS: DAPI staining positive cells were observed around the cerebral infarcted area in the ADSC group. The expressions of Neuritin, NF200 were higher, but GFAP was lower than that of the MCAO group at 7 d, 14 d and 28 d (P<0.05). CONCLUSION: The transplantation of ADSC can induce regeneration and repairment of impaired neuronal axons in rat brain after cerebral ischemia, partly by inhibiting the expression of GFAP and enhancing the expressions of Neuritin, NF-200 in the brain.

Effects of the Chinese Yi-Qi-Bu-Shen Recipe extract on brainstem auditory evoked potential in rats with diabetes.

This study was designed to investigate the effects of the Chinese Yi-Qi-Bu-Shen Recipe (YB) on brain stem auditory evoked potential (BAEP) in diabetic rats and on the protection of the diabetic rat brain's functional lesion. Thirty-three male rats were randomly divided into three groups: the normal control group (NC), the diabetic group (DM), and the diabetic rats treated with YB group (DM+YB). Blood glucose and body weight were measured every three weeks. After six weeks, the serum insulin, blood biochemical indices, superoxide dismutase, malondialdehyde, monoamine neurotransmitters, and BAEP were measured. Compared with the NC group, the waves III, V PLs, and the I-III, I-V IPLs of BAEP in the DM group were significantly delayed (all P<0.05). However, YB-treated diabetic rats maintained a normal brainstem function over the experimental period. Compared with the NC group, the waves I, III, V PL, and waves I-III, III-V and I-V IPLs of BAEP in the DM+YB group were very close (all P>0.05). On the other hand, compared with the DM group, the III, V PLs and the I-III, I-V IPLs of BAEP in the DM+YB group were significantly improved. It was discovered that the central conduction time of rats with diabetes had a close correlation with serum insulin, blood glucose, malondialdehyde, and insulin resistance index. Our results suggest that YB extract has a beneficial effect in preserving the brain's electrophysiological function in diabetic rats, likely through its antihyperglycemic activity, ability to reduce insulin resistance, and antioxidant activity.

[Effects of adipose-derived stem cells transplantation on the neuronal apoptosis and the expression of Bcl-2 and caspase-12 in the brain post focal cerebral ischemia in rats].

AIM: To investigate the effects of adipose-derived stem cells (ADSCs) transplantation on neuronal apoptosis in the brain after focal cerebral ischemia in rats. METHODS: 72 male adult Sprague-Dawley rats were randomly divided into 4 groups: Sham-operated group , Middle cerebral artery occlusion (MCAO) group, Vehicle group and ADSC-treated group (n=18). MCAO model was established with the modified Longa's method. One day after right MCAO, 30 µL of cell suspension containing 1×10(6); cells were injected into the lateral ventricle of ADSC-treated group and the same dose of PBS was given to the vehicle group. At 4 d, 7 d and 14 d after MCAO, the apoptosis of neuron was detected by terminal deoxynucleotidyl transferase-mediated DNA nick-end labeling (TUNEL) method and the expression of Bcl-2 and caspase-12 in ischemic region was detected by immunohistochemistry and RT-PCR. RESULTS: TUNEL-positive cells in ischemic region of ADSC-treated group were less than that in MCAO group and Vehicle group at 4 d, 7 d and 14 d post MCAO (P<0.05). Compared with MCAO group and Vehicle group, the expression of Bcl-2 significantly up-regulated while caspase-12 expression significantly decreased in ADSC-treated group at any time point post MCAO (P<0.05). CONCLUSION: The transplantation of ADSCs can reduce neuronal apoptosis of rats with cerebral ischemic injury partly by promoting the expression of Bcl-2 which participates in apoptotic signals after mitochondrial damage and inhibiting the expression of caspase-12 which mediates endoplasmic reticulum (ER) stress-induced apoptosis.

[Effects of the transplantation of adipose-derived stem cell on the expression of Notch1-Dll4 signaling pathway in brain of rats with focal cerebral ischemia].

OBJECTIVE: To investigate the effects of the transplantation of adipose-derived stem cells (ADSC) on the expression of Notch1-Dll4 signaling pathway in brains of rats with focal cerebral ischemia. METHODS: Sixty-five male adult Sprague-Dawley rats were randomly divided into 4 groups: sham-operated group, MCAO (occlusion of middle cerebral artery) group, ADSC-treated group and ADSC & DAPT-treated group. A permanent model of focal cerebral ischemia was established by modified Zea-Longa's method. At 24 hours post-MCAO, 1×10(6) DAPT-labeled ADSC were injected into the lateral ventricle of rats in the ADSC-treated group and the same dose of ADSC with DAPT (γ secretase inhibitor, N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester) to the rats in the ADSC & DAPT-treated group. Rats are sacrificed at 4, 7, 14 and 28 d post-MCAO. The amount of microvessels was quantified. And the levels of Notch1, Dll4 and Hes1 were detected by Western blot and immunohistochemistry. RESULTS: The density of microvessels significantly increased in the ADSC group (13.93 ± 0.50, 17.90 ± 0.62, 20.78 ± 0.80, 17.28 ± 1.65) versus the MCAO group (7.03 ± 0.22, 10.83 ± 0.63, 16.35 ± 0.54, 13.80 ± 2.38) (P < 0.05) and the ADSC + DAPT group (5.73 ± 0.30, 7.58 ± 0.52, 7.65 ± 0.45, 6.48 ± 1.47) (P < 0.05). And compared with the MCAO group (1.29 ± 0.07, 2.13 ± 0.21, 1.92 ± 0.03) and the ADSC + DAPT group (1.162 ± 0.099, 1.684 ± 0.180, 1.041 ± 0.040), the expressions of Notch1, Dll4 and Hes1 proteins were significantly up-regulated at 14d in the ADSC group (2.52 ± 0.22, 4.52 ± 0.36, 2.62 ± 0.05) (P < 0.05). CONCLUSION: The transplantation of ADSC can improve angiogenesis by up-regulating the post-MCAO expression of Notch1-Dll4 signaling pathway in rats.