Aim To investigate whether the effect of Aβ25-35 on Bcl-2 and Bax gene transcription through DNA methylation in SH-SY5Y cell.Methods Differ-ent concentrations of Aβ25-35 (0, 25, 50 μmol? L-1 ) were treated with SH-SY5Y cells for 48 h or 72 h in vitro.The optimal concentration and time of Aβ25-35 in-duced SH-SY5 Y apoptosis were determined by MTT method.Protein expression levels of Bcl-2 and Bax of Aβ25-35-treated groups were determined by Western blot.Real time PCR was used to detect the mRNA lev-els of DNA methyltransferase including DNMT 1 , DN-MT3a, DMT3b, MeCP2. Methylation specific PCR ( MSP) was used to analyze the effect of Aβ25-35 media-ted Bcl-2 and Bax gene promoter methylation .Results 25 μmol? L-1 Aβ25-35 was exposed to SH-SY5Y cells for 72 h, MTT assay showed that cell viability was (68.49 ±9.83 )%, which was significantly reduced compared with the control group ( P <0.05 ) , indica-ting AD cell apoptosis model was successfully estab-lished.Bcl-2 expression of Aβ25-35-treated group was significantly reduced compared with the control group , on the contrary , the expression of Bax was significantly increased .Real-time PCR results showed that com-pared with the control group , DNMT1, DNMT3a, DMT3b, MeCP2 mRNA levels of the Aβ25-35-treated groups had no significant difference ( P>0.05 ); MSP results showed that Bcl-2 and Bax unmethylated ampli-fication was positive , methylated amplification was neg-ative in control group , Bcl-2 and Bax unmethylated amplification was positive and methylated amplification was negative in Aβ25-35-treated group.Conclusion DNA methylation of Bcl-2 and Bax gene promoter are not affected during Aβ25-35 induced SH-SY5Y cell ap-optosis .

BACKGROUND:There are several routes for stem cel transplantation;however, it is stil unable to determine which one is the best. As for the different individuals with brain injury, the type of transplanted cel s, transplantation route and time wil affect the therapeutic effects.
OBJECTIVE:To investigate the effect of bone marrow mononuclear cel s transplanted via different approaches on neurological function of rats with traumatic brain injury.
METHODS:Bone marrow mononuclear cel s of rats were administered gradient centrifugation with Ficol lymphocyte separation medium, and were labeled with CFDA-SE in vitro as standby. Rat models of traumatic brain injury were established by the method of freefal . After successful establishment of rat models, bone marrow mononuclear cel s labeled with CFDA-SE were immediately transplanted into rats via injured area, lateral ventricle and internal carotid artery. One control group was designated for each transplantation route (bone marrow mononuclear cel s were replaced with the same volume of DMEM). The degree of neurological deficits was evaluated using mNSS scores at different time points after treatment. The brain tissue was harvested after the last neurobehavioral evaluation. The survival and migration of bone marrow mononuclear cel s in the injured area were observed under an inverted fluorescent microscope.
RESULTS AND CONCLUSION:At 7, 10, and 14 days after treatment, the mNSS scores of rats in al groups were al lower than those at 1 and 3 days (P<0.05). At 7 and 10 days, the mNSS scores of rats in the internal carotid artery transplantation group were significantly lower than those in the control group (P<0.05). At 14 days after treatment, the number of fluorescence-labeled cel s of rats in the internal carotid artery transplantation group was greater than that in the other groups (P<0.05) and these labeled cel s were widely distributed. The results demonstrate that the neurological function of rats can be improved by transplanting bone marrow mononuclear cel s via the internal carotid artery, and a large number of transplanted cel s can survive and migrate in the injured area.

Aim To study the protective effect of CDPS on acute aging mouse model induced by D-galactose (D-gal) and its mechanism.Methods (1) The acute aging mouse model was induced by D-gal.After CDPS (25、50、100 mg·kg-1) treatment, the improving effect on learning and memory in mice was examined in vivo.(2) We also established the aging model on PC12 cells in vitro.After CDPS treatment (150、200 mg·L-1), the level of p-CREB in the nucleus was detected by Western blot, and the content of cAMP, PKA and brain derived neurotrophic factor (BDNF) levels were examined by the Elisa kits.Moreover, cAMP, PKA and BDNF were detected in PC12 cells under the condition that H89, the inhibitor of PKA, co-cultured with PC12 cells after CDPS treatment.(3) The UPLC/Q Exactive MS method was developed for determining the concentration of glutamic acid, dopamine and norepinephrine, which secreted in PC12 cells after CDPS treatment.Results (1) In vivo, CDPS significantly improved the memory impairment in aging mice induced by D-gal in the Morris assay.(2) In vitro, CDPS could significantly increase the expression of p-CREB (P<0.05), PKA, cAMP and BDNF (P<0.05).The H-89 abolished the increase of p-CREB (P<0.05), PKA, cAMP and BDNF (P<0.05) in PC12 aging cells induced by D-gal after CDPS treatment.(3) CDPS increased the release of dopamine, norepinephrine, and glutamate secreted in PC12 cells.Conclusion CDPS could significantly improve the learning and memory ability on aging mouse model in vivo, and reversed the damage in PC12 cells induced by D-gal by activating cAMP/PKA/CREB signal cascade, increase the expression of BDNF, and increasing modestly the release of excitatory neurotransmitter.

OBJECTIVE:To study the improvement effect of lanthanum hydroxide on chronic renal failure (CRF) hyperphos-phatemia in rats. METHODS:CRF hyperphosphatemia rat model were induced and then randomly divided into model group,lan-thanum carbonate group [0.3 g/(kg·d)],calcium carbonate group [4.2 g/(kg·d)] and lanthanum hydroxide high-dose,medium-dose and low-dose groups [1.5,1,0.5 g/(kg·d)] with 10 rats in each group. They were given adenine 0.2 g/(kg·d)intragastrically in the morning,and then given relevant medicine intragastrically in the afternoon;a week later,they stopped taking adenine but con-tinued to take relevant medicine for 22 d. 10 normal rats were selected as normal control group. General examination was conduct-ed,and renal coefficient,serum contents of calcium,phosphorus,PTH,creatinine(Scr)and usea nitrogen(BUN)were detected after last medication as well as renal pathological change. RESULTS:Compared with normal control group,model group showed CRF sign,renal coefficient,the contents of phosphorus,PTH,Scr and BUN were increased,while the content of calcium was de-creased(P<0.01);renal section showed obvious pathological characteristics. Compared with model group,CRF sign of rats were improved in lanthanum carbonate group,calcium carbonate group and lanthanum hydroxide groups. The renal coefficient (except for lanthanum hydroxide high-dose group),serum contents of phosphorus(except for calcium carbonate group),PTH(except for lanthanum hydroxide low-dose group and calcium carbonate group),Scr(except for lanthanum hydroxide low-dose group and calci-um carbonate group)and BUN were all decreased,while serum content of calcium and calcium-phosphorucs product(only in calci-um carbonate group)was increased(P<0.05 or P<0.01). There was no statistical significance in other difference. The renal sec-tion pathological characteristics were improved. CONCLUSIONS:Lanthanum hydroxide can improve renal function and reduce the level of serum phosphorus in CRF hyperphosphatemia model rats.

Objective To investigate the expression and correlation of NKG2D and sMICA in lung cancer patients. Methods By collecting 30 lung cancer patients as the test group,and taking 30 healthy volunteers as the contrast group, the expression of NKG2D and sMICA in the two groups were examined separately by FACS and ELISA method. Results The expressions of NKG2D in the two groups were (81.56±8.78) %, (85.63±6.62) %. The lung cancer patients were high remarkable. There was a significant difference between the two groups (P <0.05). The expression of sMICA in the two groups were (354.13 ±80.575) pg/ml,(216.53±48.175) pg/ml. The lung cancer patients were low remarkable. There was a significant difference between the two groups (P <0.01). There was a significant relation between the two groups (r =-0.349, P =0.006). Conclusion The expression of NKG2D and sMICA may provid one of the immune targets for diagnosing that can forecast the immune state and malignant metastasis of the lung cancer patients. The significant relation between NKG2D and sMICA may take on main role in the immune escaping of tumor. It may provide the suitable target of the patients for tumor organisms and immune treatment.

Objective To Explored the relative factors which caused the extubation failure in neurological intensive care unit (NICU).Methods It was a retrospective study.40 cases of patients who met the criteria,were brought into statistical analysis.They were admitted in NICU in Nan Fang Hospital from December 2008 to February 2011.The name,sex,age,diagnosis,respiratory parameters,24 hours discrepancy quantity,sputum,and Glasgow Coma Scale,Full Outline of UnResponsiveness Scale were recorded.SPSS 13.0 was used as statistic software.P ＜ 0.05 was considered statistically significant.Results Both in extubation successful and failure groups,GCS and Four were significantly different (all P ＜ 0.05).Howerer,there were no statistically significant in the other factors.There were significantly differences between GCS and Four in predicting extubation results (P =0.012).Logistic multiple regression showed that Four and GCS grade were predictive factor of extubation failure (P =0.041).Conclusions The result suggests that it is statistically significant to use GCS and Four as factors to predict extubation results.It can be widely used to help medical personnels monitoring the changes of patients'clinical conditions,judging prognosis,and making treatment plan in NICU.Wether other factors would effect the extubation results,more prospective,randomized controlled studies were needed.

BACKGROUNDThe abnormalities of coronary arteries, though rare and sometimes benign, may first present clinically as myocardial infarction or sudden death. Multi-detector computed tomography (MDCT) is a non-invasive test that is highly suitable for detecting these anomalies. The study aimed to review the 64-MDCT appearance of the coronary artery anomalies in 66 patients and to discuss the clinical importance of these anomalies.

METHODSIn 6014 consecutive patients examined over 12 months by 64-MDCT for the study of coronary artery disease, 66 were diagnosed for coronary artery anomalies. All patients were symptomatic for one or more of the following diseases: chest pain, dyspnoea, palpitations, arrhythmia and myocardial infarction. Nine patients had undergone a coronary angiography. All the CT images were evaluated by two radiologists and one cardiologist. The right coronary artery (RCA) and the conus branch arising separately, myocardial bridging and duplication of arteries were not analysed in our study.

RESULTSThe incidence of coronary artery anomalies found in our study group was 1.097%. In the selected patients, seven different types of coronary anomalies were found by 64-MDCT examination. The high takeoff, origin of the coronary artery from the opposite or noncoronary sinus with an anomalous course, and coronary artery fistula were the three common forms of anomalies (n = 16, 18 and 16, respectively). Compared with the results of the coronary angiography, the number of the drainage sites of two coronary artery fistula was less in MDCT images (3 small sites in total). In all cases, coronary artery computed tomography angiography (CTA) technique was able to recognize the origin of the coronary artery, its three-dimensional course and its spatial relationship with the adjacent structures. Conventional coronary angiography in two cases, however, was unable to provide sufficient information for correct and complete diagnosis.

CONCLUSIONSIn conclusion, the study showed that 64-MDCT, especially the volume rendering technique (VRT), may be useful for the assessment of complex variations, even if the conventional angiography may not be sufficient. It may be considered as the first-choice imaging modality when an anomalous coronary artery is suspected.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Coronary Vessel Anomalies ; diagnostic imaging ; Female ; Humans ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods ; Young Adult

OBJECTIVETo determine the feasibility and assess the validity of noncontact endocardial mapping to guide ablation of hemodynamically unstable or nonsustained ventricular tachycardia (VT).

METHODSNoncontact mapping permitted individual-beat analysis of ventricular arrhythmias. Three-dimensional electroanatomical mapping allowed detailed reconstruction of a chamber geometry and activation sequence. Eighteen hemodynamically unstable or nonsustained VTs were induced (cycle length: 336 ms +/- 58 ms) in 17 patients and mapped by noncontact mapping using an EnSite 3000 system performed for the guidance of catheter ablation.

RESULTSThree patients were mapped during premature ventricular complexes (PVCs) because sustained VT could not be induced. Analysis of the archived noncontact activation maps was performed to identify the exit site and/or the diastolic pathway of the VT reentry circuit. The endocardial exit sites 10 ms +/- 16 ms before QRS were defined in 9 right ventricular outflow tract (RVOT) and 5 ischemic VTs. The diastolic pathway was identified in 5 ischemic VTs. The earliest endocardial diastolic activity preceded the QRS onset by 60.1 ms +/- 42.6 ms. The earliest activation sites were identify in 3 patients with nonsustained VTs or PVCs. Radiofrequency current was applied around the exit site or to create a line of block across the diastolic pathway. Catheter ablation was performed in 17/18 (94%) VTs and 15/17 (88%) VTs was successfully ablated. Two (67%) of the three patients with non-sustained VTs were mapped and successfully ablated during PVCs. Catheter ablation was not performed in 1 patient (peri-Hisian VT) and was unsuccessful in 2 patients.

CONCLUSIONNoncontact endocardial mapping is able to be used to guide ablation of untolerated or nonsustained VTs.

Adult ; Catheter Ablation ; methods ; Electrophysiologic Techniques, Cardiac ; Feasibility Studies ; Female ; Humans ; Male ; Middle Aged ; Tachycardia, Ventricular ; physiopathology ; surgery

To generate transgenic mice in which both hygromycin (hyg) and neomycin (neo) resistance genes are expressed in murine fibroblast cells (MEFs), which are required for conditional gene knock-out and screening of drug resistant ES cell clones. To construct HygR-neoR expression vector, pTK-hygR-pA and PGK-neoR-pA were cloned into pBluescript vector. DNA fragments of tandem genes ( 4245bp ) were prepared by Kpn I and Xba I digestion and transgene was microinjected into pronucleus of zygotes to generate transgenic mice. Transgenic mice were identified by PCR and Southern blot; expression of hygR and neoR gene transcripts were detected by RT-PCR. 7 founder mice carrying hyg-neo resistant genes were obtained and 6 transgenic mouse lines were successfully established. The hygR and neoR gene transcripts were detected in the liver and/or ovary of transgenic mice from hn30, hn33, hn66 and hn67 mouse lines. In MEFs isolated from the mice of line hn66 and hn30, expression of hyg and neo resistant genes was also detectable. Transgenic mouse lines expressing two anti-drug genes have been established. The hyg and neo resistant gene transcripts were detected in the MEFs of two transgenic mouse lines.
Animals ; Cinnamates ; pharmacology ; Drug Resistance, Multiple ; genetics ; Fibroblasts ; metabolism ; Hygromycin B ; analogs & derivatives ; pharmacology ; Mice ; Mice, Transgenic ; Neomycin ; pharmacology ; Transgenes ; genetics

PURPOSE: Breast cancer (BC) is one of the most common malignant tumors, affecting a significant number of women worldwide. MicroRNAs (miRNAs) have been reported to play important roles in tumorigenesis. The aim of this study was to determine the roles of miR-182-5p in BC progression. MATERIALS AND METHODS: The expressions of miR-182-5p and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) were measured in BC tissues and cells by quantitative real-time polymerase chain reaction or Western blot. Cell proliferation and invasion were detected by cell counting kit-8 assay and trans-well assay, respectively. The interaction between miR-182-5p and PTEN was probed by bioinformatics analysis, luciferase activity, and RNA immunoprecipitation. A murine xenograft model was established to investigate the role of miR-182-5p in BC progression in vivo. RESULTS: An abundance of miR-182-5p was noted in BC tissues and cells. High expression of miR-182-5p was associated with poor survival. Abrogation of miR-182-5p inhibited cell proliferation and invasion in BC cells. Interestingly, PTEN was indicated as a target of miR-182-5p, and its restoration reversed miR-182-5p-mediated promotion of proliferation and invasion of BC cells. Moreover, depletion of miR-182-5p suppressed tumor growth via up-regulating PTEN expression in the murine xenograft model. CONCLUSION: MiR-182-5p exhaustion blocked cell proliferation and invasion by regulating PTEN expression, providing a novel therapeutic avenue for treatment of BC.
Blotting, Western ; Breast Neoplasms ; Breast ; Carcinogenesis ; Cell Count ; Cell Proliferation ; Chromosomes, Human, Pair 10 ; Computational Biology ; Female ; Heterografts ; Humans ; Immunoprecipitation ; Luciferases ; MicroRNAs ; Real-Time Polymerase Chain Reaction ; RNA