AIM:To explore the role of microRNA-181b (miR-181b) in ischemic injury and autophagy pro-tein 5 (Atg5) levels of mice .METHODS:Oxygen-glucose depletion (OGD) model in N2A cells to mimic ischemic in-jury in vitro was established .A middle cerebral artery occlusion ( MCAO) model to mimic ischemic injury in vivo was also induced in mice.The N2A cell apoptosis after OGD was assessed by in situ cell death detection kit.The Atg5 and caspase-9 expressions were determined by Western blotting .Luciferase reporter assay was performed to identify the direct binding of miR-181b with 3’-UTR of Atg5 mRNA.RESULTS:The alteration of miR-181b expression level by transfection with pre-miR-181b or anti-miR-181b significantly affected N2A cell apoptosis (P<0.05).Accordingly, the changes of miR-181b levels significantly altered the protein level of Atg 5 ( P<0.05 ) .Co-transfection of the luciferase reporters with pre-miR-181b or anti-miR-181b resulted in the inhibition or enhancement of the luciferase activities of luciferase expressing plasmid containing 3’-UTR of Atg5 mRNA (P<0.05).In addition, the miR-181b antagonist significantly reduced the cleaved caspase-9 levels in cerebral ischemic cortex of the mice after MCAO ( P<0.05 ) .CONCLUSION: Down-regulation of miR-181b plays an important role in ischemic injury of mice through regulating Atg 5 protein level.

Objective To investigate the effect of silodosin,a selective alpha1 a-adrenoceptor antagonist on a rat model of testosterone-induced benign prostatic hyperplasia (BPH) and its mechanisms.Methods The rats were divided into three groups:control,testosterone-induced BPH,and silodosin +BPH groups.BPH was induced by subcutaneous injection of testosterone [20 mg/(kg · d)] for 4 weeks.Meanwhile silodosin + BPH groups rats were administered silodosin 4 weeks [100 μg/(kg · d)].After 4 weeks,all animals were sacrificed to examine the blood biochemical profiles,prostate volume,weight,histopathological changes,and epidermal growth factor receptor (EGFR) and B-cell chronic lymphocytic leukemia (CLL)/lymphoma 2 (BCL-2) protein expressions.Results Each group showed an increase compared to their initial body weight;however,differences in weight change between groups were not significant (P ＞ 0.05).The BPH group displayed lower glucose levels than the control group.The serum levels of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) were not significantly different among groups (P ＞ 0.05).The group treated with silodosin showed significantly lesser prostate size and weight than the testosterone-induced BPH group [volume:(0.93 ± 0.14) cm3 vs (1.75 ± 0.15)cm3,P ＜0.01;weight:(0.97 ±0.06)g vs (1.30±0.05)g,P ＜0.01].In addition,silodosin decreased the expressions of EGFR and BCL-2 in prostate tissues (P ＜ 0.05).Conclusions These results suggest that silodosin suppress the development of BPH by inhibiting the expressions of EGFR and BCL-2.

Objective To explore the change and function of CD 4 +CD 25 + and CD 8 +CD 28 -T regulatory cells in peripherel blood monocyte cell (PBMC), spleen and colon in rat experimental colitis induced by trinitrobenzenesulfonic acid (TNBS) . Methods The rat model of experimental colitis was established by enema with TNBS/ethanol. The proportion of CD 4 +CD 25 + and CD 8 +CD 28 - T regulatory cells in PBMC, spleen and colon was determined by flow cytometry at the first and third weeks after establishing model, respectively. Results The model of experimental colitis in rats was successfully established(n=15). Compared with control group, the proportion of CD 8 +CD 28 -T regulatory cells significantly increased in colon at the first week after establishing model [(12.7?5.4)% versus(3.87?3.7)%,P

Objective To clone the 5′ non-coding region (NCR) of human interferon-?-inducible protein 10(IP-10), and to identify the transcriptional activity of IP-10 promoter induced by lipopolysaccharide (LPS) in human umbilical vein endothelial cells (HUVEC). Methods Genomic DNA of lymphocytes was isolated from the human blood. With above DNA as the template, the 5'NCR of human IP-10 was amplified by nest polymerase chain reaction (PCR) method. Then, the IP-10 promoter was cloned into luciferase reporter vector, pGL3. The recombined vector was transfected into HUVEC, and then the activity of the luciferase was determined after the cells were stimulated by LPS. Results Human IP-10 promoter was obtained and the pGL3/IP-10 was successfully constructed. Moreover, the activity of luciferase driven by human IP-10 promoter was observed to obviously increase in the HUVEC stimulated by LPS. Conclusion We successfully cloned human IP-10 promoter, constructed luciferase reporter vector driven by the human IP-10 promoter, and confirmed that high transcriptional activity of human IP-10 promoter was induced by LPS in HUVEC. The results supplied an experimental base for the further study of the transcriptional regulation of human IP-10.

In view of the fact that medical inspection equipment sold in the domestic market is mainly imported from abroad and very expensive, we developed a full-automatic fluorescence analyzer in our center, presented in this paper. The present paper introduces the hardware architecture design of FPGA/DSP motion controlling card+PC+ STM32 embedded micro processing unit, software system based on C# multi thread, design and implementation of double-unit communication in detail. By simplifying the hardware structure, selecting hardware legitimately and adopting control system software to object-oriented technology, we have improved the precision and velocity of the control system significantly. Finally, the performance test showed that the control system could meet the needs of automated fluorescence analyzer on the functionality, performance and cost.
Automation, Laboratory ; Equipment Design ; Fluorescence ; Software

Objective To explore the application and value of three-dimensional reconstruction technology in diagnosis and surgical therapy of ground-glass opacity.Methods We analyzed 188 cases with GGO during October 2012 and October 2014 in Shanghai First People’s Hospital affiliated to Shanghai Jiao tong University School of Medicine respectively.Several relative parameters based on three-dimensional reconstruction were selected to differentiate GGO pathology .By ROC curve anal-ysis, optimal cut-off points were determined.Volume-doubling time(VDT) was calculated among 104 cases which had been following up at least for three months and compared among AAH, MIA and IAC.Results Cases’ mean age was 56.66 ± 9.673years(ranged from 27 to 82 years) with 67 males and 121 females.The diameter(P<0.001), total volume(TV)(P<0.001), the maximum CT attenuation(MAX)(P<0.001) and standard deviation of distribution of CT attenuation within the whole GGO(STD)(P=0.015) of malignant group were significant larger than those of benign ones except for average CT at-tenuation(AVG)(P=0.094).The AUC were just 0.64-0.80 when differentiated benign group from AIS, benign group from MIA and AAH from MIA in comparison to 0.70-0.95 when it comes to differentiating benign group and AAH from IAC .Ad-ditionally, the optimal cut-off points were 13.5-15.5 mm in diameter, 400-600 mm3 in TV, -110 -20 Hu in MAX and 120-160 in STD.The mean VDT was 865.00 ±111.33, 464.67 ±44.40 and 238.36 ±76.71 for AAH, MIA and IAC re-spectively(AAH/MIA P<0.001, MIA/IAC P=0.003).Conclusion The diameter, TV, MAX and STD of GGO based on three-dimensional reconstruction could discriminate GGO availably .Combined utilization of HRCT and three-dimensional re-construction has a valuable significance in diagnosis and treatment of GGO .

Gallibacterium anatis,a member of the Pasteurellaceae family,constitutes a part of the normal micro-flora of the upper respiratory tract and the lower genital tract in chickens.However,increasing evidence indicate that G.anatis is also associated with a wide range of pathological changes,particularly in the reproductive organs,which leads to decreased egg production,lowered animal welfare and increased mortality.As a recently defined opportunistic pathogen limited focus has been placed on the pathogenesis and putative virulence factors permitting G.anatis to cause disease.One of the most studied virulence determinants is a large RTX-like toxin (GtxA),which has been demonstrated to induce a strong leukotoxic effect on avian macrophages.A number of fimbria of different sizes and shapes,particularly the F17-1ike fimbriaes,appear to be common in a diverse selection of G.anatis strains.The capsular material expressed possibly involved in serum resistance,metalloproteases capable of degrading immunoglobulins,hemagglutinins,and all factors which may promote biofilm formation are likely linked to the virulence of G.anatis.This review summarized the putative virulence factors described for this bacterium to date.

Objective To investigate any therapeutic effect of combining hyperbaric oxygen with mesenchymal stem cells (MSCs) in treating traumatic brain injury (TBI).Methods Eighty healthy adult rats were randomly divided into a control group,a hyperbaric oxygen group,a stem cell group and a combination group,each of 20.TBI was introduced into the rats of all 4 groups.Twenty-four hours after the modelling,the hyperbaric oxygen group received hyperbaric oxygen therapy,the stem cell group received MSCs transplantation,the combination group was given the hyperbaric oxygen therapy an hour after the MSCs transplantation,while the control group was not given any treatment.All of the rats were evaluated using neurological severity scores (NSSs) after the modeling and again after the treatment.They were then sacrificed for HE staining and the expression of nuclear factor kappa B (NF-kB) and brain-derived neurotrophic factor (BDNF) were observed.Results On the 3rd,5th,10th and 20th day after the modeling,the average NSS of the combination group was significantly lower than those of the other three groups.However,the average NSS of the combination group on the 20th day was significantly superior to that on the 3rd and the 5th days.Compared with the control group,the edema of brain cells was less severe in the other 3 groups.The average expression of NF-kB and BDNF in the combination group was significantly higher than in the other 3 groups on the 3rd,5th,10th and 20th day after the modeling.Conclusion Hyperbaric oxygen,especially long term treatment combined with stem cell transplantation,can significantly improve nerve function in the brain after trauma,relieve inflammation and edema in and around the damaged area,and promote the expression of the NF-kB and BDNF.

AIM: To construct prokaryotic expression vector of His-tagged human IP-10 for further study of its biological function in the inflammatory response. METHODS: The coding sequence of IP-10 lacking signal peptide was amplified from human lung cDNA library by polymerase chain reaction (PCR) and the fragment was cloned into pET-14b plasmid for the construction of His-tagged fusion protein expressing vector, pET-14b/IP-10. After being identified by enzyme digestion and sequencing, the recombinant vector was transformed into a strain of E. coli, BL21 (DE_3). The expression of His-tagged fusion protein was induced with IPTG and purified with Ni+-NTA affinity chromatography. Then the chemotactic activity of IP-10 was determined by transwell migration assay on THP-1 cells. RESULTS: The construction of pET-14b/IP-10 recombinant vector was proved by enzyme digestion and sequencing. The fusion protein IP-10, which was purified by a routine Ni+ affinity method, had an activity on the induction of cell migration of THP-1. CONCLUSION: We successfully construct IP-10 fusion protein expressing vector and get the fusion protein with high bioactivity, which provides essential materials for the future studies on IP-10.