Objective To assess cerebrovascular reserve ( CVR) function in patients with internal border zone infarction(IBZI) induced by severe middle cerebral artery (MCA) stenosis, and investigate the impact on progression and outcome of the disease .Methods A total of 84 patients with unilateral severe MCA stenosis were selected . Hypercapnia was induced by holding breath .The change of blood flow velocity in MCA was measured by transcranial Doppler ( TCD ) to calculate CVR .According to CVR , patients were divided into impaired regional CVR group ( CVR <10%) and normal CVR group ( CVR ≥10%) .The NIHSS was used to evaluate neurological function in both groups within 14 d, and mRS was used to evaluate acute stage outcome of the patients at discharge .All the patients were followed-up for 6 months, the incidences of recurrence , complications and mortality in the two groups were analyzed .Results The incidence of progressive cerebral infarction in the impaired regional CVR group (67.39%) was significantly higher than that in the normal CVR group (42.11%) ( P<0.05).The impaired regional CVR group showed higher proportion of patients with poor clinical outcome at discharge ( mRS≥3 ) (63.04%)compared to the normal CVR group (36.84%) (P<0.05).In the followed-up 6 months, the incidences of recurrence and complications were 34.78% and 45.65% respectively in the impaired regional CVR group , they were significantly higher than that in normal CVR group (15.79%,23.68%)(P<0.05).The overall mortality rates did not differ significantly between the two groups ( P>0.05 ) .Conclusion Impaired regional CVR may be predictive of subsequent progressive cerebral infarction and poor clinical outcomes in patients with IBZI induced by severe MCA stenosis .

Objective To promote the development of precision medicine and gene sequencing technologies in China by analyzing the innovation situation in gene sequencing technologies.Methods An innovation situation research analysis frame was established with scientific literature and patent information as its basic data.The innovation situation in gene sequencing technologies was analyzed in aspects of scientific research and technical R & D.Results The research on gene sequencing technologies and technical R & D were focused on tumor diagnosis and treatment,plant gene sequencing,identification of HIV and gastrointestinal flora.Single cell genome amplification technology has greatly promoted the development of gene sequencing technologies.Conclusion The level of gene sequencing technologies and technical R & D in China is approaching to that in the world.

In this paper, we designed and evaluated a duplex detection strategy for microRNAs (miRNAs) using universal probe-based target-triggered double hybridization and fluorescent microsphere-based assay system (xMAP array). In the absence of target miRNA, reporter DNA cannot hybridize stably with the immobilized capture DNA due to its low melting temperature. Only after adding target miRNA, can reporter probe hybridize with capture probe to form a stable three-component complex. This targettriggered stable hybridization makes this method possible for highly selective and sensitive detection of multiple miRNAs. We exemplified a quantitative detection of duplex miRNAs with a limit of detection of 40 pM. The xMAP array platform holds the potential of extending this approach to simultaneous detection of up to 100 miRNA targets. Considering the simplicity, rapidity and multiplexing, this work promised a potential detection of multiple miRNA biomarkers for early disease diagnosis and prognosis.

Background and purpose:Inhibitor of apoptosis-stimulating protein of p53 (iASPP) is one of the ASPP family. It binds to p53 to inhibit the transcriptional activity of p53-target genes and cell apoptosis, which is asso-ciated with tumor formation. Previously, we found a new subtype of iASPP, iASPP splice variant (iASPP-SV), which is a nuclear protein containing 407 amino acid residues and can bind to p53, inhibiting p53 transcriptional activity. However, the relationship of iASPP-SV and breast cancer is still obscure. Therefore, the purpose of this research was to study the role of iASPP-SV on breast cancer tumorigenesis and progression.Methods:5’-rapid ampliifcation of cDNA ends (RACE) was used to identify the 5’-end of iASPP-SV mRNA in MCF-7 cells. HEK 293 cells were transfected with pFLAG-iASPP-SV and pFLAG-iASPP (828). Then Western blot was used to identify whether endogenous iASPP-SV was expressed in HEK 293 cells and 8 types of human tumor cell lines. This study established the stable clones of NIH 3T3 expressing FLAG-iASPP-SV and FLAG-iASPP (828). Cell proliferation assay, colony formation and soft agar colony formation assay were used to identify whether iASPP-SV and iASPP (828) can promote cell proliferation and iASPP-SV is an oncogene. Real-time lfuorescent quantitative polymerase chain reactive (RTFQ-PCR) was used to de-tect the levels of iASPP-SV and iASPP (828) mRNA in primary breast cancers. Luciferase assays were used to identify the relationships between iASPP-SV, iASPP (828), p53 and NF-κB p65.Results:The study identiifed that iASPP-SV was encoded by previously reported NF-κB p65 subunit (RelA)-associated inhibitor (RAI), and endogenously expressed in many human cancer cell lines. Analysis of cell proliferation, colony formation assay and soft agar assay for colony formation identiifed that similarly to iASPP (828), iASPP-SV promoted tumor cell proliferation and acted as an onco-gene. RTFQ-PCR result showed that the median values of iASPP-SV and iASPP (828) in breast cancers with wild-type p53 were more signiifcantly over-expressed than those of mutant p53. Luciferase assays showed that iASPP-SV and iASPP (828) could suppress NF-κB p65 transcriptional activity. Thus iASPP family may participate in the regulation of p53 and NF-κB activity, which imply that iASPP perhaps shows pro- or anti-survival activities when it interacts with different proteins.Conclusion:These ifndings indicate that iASPP-SV may be a potential target for breast cancer thera-py.

Objective To observe the effects of Okinawa Habu apoxin protein-1 (OHAP-1) on the proliferation inhibition of rat C6 glioma cells and its mechanisms. Methods MTT colorimetric analysis was used to measure the inhibitory effect of OHAP-1 with different doses(2.5,5.0,and 10.0 mg?L-1) on C6 glioma cells .RT-PCR was used to evaluate the mRNA expressions of bcl-2 and bax genes.The activity of superoxide dismutase (SOD) and the level of maleicdialdehyde (MDA) in the C6 glioma cells were also examined. Results The proliferation of C6 glioma cells was significantly inhibited by different doses of OHAP-1(2.5,5.0,and 10.0 mg?L-1).The inhibitory rate were 49.77%,67.65%,and 76.42%,respectively.The inhibitory rate in 2.5,5.0, and 10.0 mg?L-1 groups were higher than that in control group(P

ObjectiveTo explore the changes of astrocytes in vitro on the expression of Cyclin D1 under normal and hypoxia/reoxygenation conditions.MethodsThe primary astrocytes were isolated from the cortex of SD fetal rats(less than 24 hours) and identified by immunosytochemical method with anti-GFAP antibody after 3 passages.Astrocytes of passage 3 were divided into normal group ( control group),hypoxia/reoxygenation group ( H/R 37℃ ),and mild hypothermiaintervention group( H/R 32℃ )separately.Astrocytes from the later tow groups were reoxygenated with 4,12,and 20 hours separately after exposed to hypoxia conditions for 8 hours.Trypan blue staining was employed to detect the survival rates and immunofluorescence,western-blot were used to analyse the expressin of Cyclin D1 of of astrocytes of different groups and time points.Results 1.The GFAP positive astrocytes from passage 3 exceeded 95 ％.2.With regard to morphology and survival rates,there is no difference between astrocytes of normal and hypothermia groups after 8 hours exposure to hypoxia conditions.Reoxygenation could obviously rise astrocytes mortality with time went by ( H/R 37 ℃ group:12.87 ± 2.76 ( R4 ),31.55 ± 3.00 ( R12 ),46.40 ±8.50(R20) ;H/R 32℃ group:6.77 ± 1.53( R4),15.97 ±4.00(R12),28.33 ±5.69(R20) ;all P＜0.05).3.Immunofluorescence and western-blot revealed that reoxygenation increased Cyclin D1expression markedly,which was proportional to the duration of reoxygenation.Mild hypothermia could reduce Cyclin D1 expression of astrocytes severely under reoxygenation condition.ConclusionCyclin D1 expression can be regarded as a sensitive index of damage to astrocytes caused by hypoxia/reoxygenation conditions.

BACKGROUND:In recent years, chitosan-based thermosensitive hydrogel, as scaffold materials, have received more and more attentions in the field of tissue repair because of good biocompatibility, biodegradability and drug-sustained release.
OBJECTIVE:To explore the directed differentiation and growth of rat bone marrow mesenchymal stem cells on the quaternary chitosan thermosensitive hydrogel scaffold and to look for more ideal tissue engineering materials for the treatment of nervous system damage.
METHODS:The thermosensitive hygrogel scaffold was prepared using hydroxypropyltrimethyl ammonium chloride chitosan (HACC) andβ-glycerophosphate (β-GP). The spatial structure of scaffold was observed by scanning electronic microscope. Effect of leaching liquor from the HACC/β-GP scaffold on the viability of bone marrow mesenchymal stem cells was detected by (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The albumin from bovine serum was combined with the scaffold, and the slow-release effect of the scaffold was detected by ultraviolet absorption spectrometry. Bone marrow mesenchymal stem cells were incubated onto the compound scaffold at 3 passages. The adhesion, growth and differentiation of bone marrow mesenchymal stem cells on the compound scaffold were observed by the scanning electron microscope. Neuron-specific enolase was detected by immunofluorescence.
RESULTS AND CONCLUSION:The porosity and thermal sensitivity of HACC/β-GP scaffold and slow-release effect of glial cellline-derived neurotrophic factor were apparent. The results of MTT showed that the compound scaffold cannot take apparent negative effects to the proliferation of bone marrow mesenchymal stem cells. After inoculation, bone marrow mesenchymal stem cells permeated the porous structure of the scaffold and adhered to the scaffold. Under the role of glial cellline-derived neurotrophic factor, bone marrow mesenchymal stem cells showed neuron-like cellmorphology and cells co-cultured with the compound scaffold expressed the marker of neurons, neuron-specific enolase. Under the role of slow-release glial cellline-derived neurotrophic factor, bone marrow mesenchymal stem cells can grow wel in vitro and differentiate into neuron-like cells on the HACC/β-GP scaffold.

Dengue virus(DENV)is an enveloped single positive-stranded RNA virus that belongs to the Flaviviridae family. It is the cause of dengue fever,life-threatening dengue hemorrhagic fever and den-gue shock syndrome. Aedes aegypti and Aedes albopictus are responsible for the wide transmission of DENV in tropical and subtropical regions throughout the world. Currently,there have been several dengue vaccines entering clinical trials,including the Sanofi Pasteur chimeric yellow fever dengue tetravalent vaccine (CYD),which has been licensed for use in some countries. However,CYD does not provide adequate pro-tection against all four serotypes of DENV and induces severe dengue diseases in young and seronegative vac-cine recipients. Therefore,a more efficacious dengue vaccine is still needed. Here,we reviewed the oppor-tunities and challenges for the development of dengue vaccines.

To investigate the cytotoxicity of the homemade peptide cationic liposome CDO14 and its efficacy of RNA interference (RNAi). MTT method was used to determine the cytotoxicity of the liposome to a human lung cancer cell line Luc-A549 that can express luciferase stably. Luciferase siRNA (Luc-siRNA) was transfected into Luc-A549 cells by CDO14. Contents of luciferase in the transfected cells were detected by luminous instrument and contents of total protein in these cells were detected by BCA method. Nude mice were inoculated with Luc-A549 cells in axilla to establish xenograft tumor model. Complexes of Luc-siRNA and the cationic liposomes were injected into the modeling mice via tail vein. Contents of luciferase in the transfected mice were detected by the whole body imaging system. The cytotoxicity of the homemade cationic liposome was similar to that of commercial liposome DOTAP, and lower than that of Lipo2000. The siRNA transfection efficacy mediated by CDO14 was higher than that mediated by DOTAP. The homemade peptide cationic liposome CDO14 is expected to serve as delivery vector in gene therapy because of its low cytotoxicity and high transfection efficiency.
Animals ; Cations ; Cell Line, Tumor ; Fatty Acids, Monounsaturated ; Genetic Therapy ; Genetic Vectors ; Humans ; Liposomes ; Luciferases ; Lung Neoplasms ; Mice ; Mice, Nude ; Peptides ; Quaternary Ammonium Compounds ; RNA Interference ; RNA, Small Interfering ; Transfection

The present paper aims to investigate the relation between characteristic parameters of transesophageal photoelectric pulse wave in descending aorta and ambulatory artery blood pressure. The chests of ten adult experimental dogs were performed to take the photoelectric pulse wave of descending aorta transesophageally. The concurrent femoral artery invasive blood pressure was recorded simultaneously. Stepwise regression analysis method was used to study the correlation efficient between characteristic parameters of descending aorta pulse wave (H, h, h/H, g/H, At, s, H(1 + ts/td), k)and invasive artery blood pressure. The characteristic parameters, k and h/H (ratio: 90% and 80%) was proved that they had good correlation with systolic pressure; and k, H and s (ratio: 90%, 80% and 70%), had good correlation with diastolic pressure; while k and H (ratio: 90% for both) had good correlation with mean pressure. The mean values of multiple correlation coefficients of the selected characteristic parameters of descending aorta pulse wave with systolic pressure, diastolic pressure and mean pressure of femoral artery were 0.871, 0.900 and 0.856, respectively. The characteristic parameters of descending aorta pulse wave had specific correlation with systolic pressure, diastolic pressure and mean pressure.
Animals ; Aorta, Thoracic ; physiology ; Blood Pressure ; physiology ; Blood Pressure Monitoring, Ambulatory ; methods ; Dogs ; Electrophysiologic Techniques, Cardiac ; methods ; Female ; Femoral Artery ; physiology ; Male ; Pulse Wave Analysis ; methods ; Regression Analysis