Objective To study the effect of adenoviral-delivered short hairpin RNA (shRNA) target against permeability glycoprotein (Pgp) as a new drug in anti-epileptic drug resistance epilepsy treatment and to evaluate its efficiency. Methods MDR Sprague-Dawley (SD) rat estrocyte model was induced by Coriaria Lactone (CL), mainly over-expressing mdrlb. To reverse the drug resistance, astrecytes were treated with constructed replication deficient adencvirus AdS-EGFP-shRNAI-U6 delivering short hairpin (shRNA) target agianst mdrlb gene. Total RNA and protein were extracted from the infected cells, mdr1 b level was detected by Quantitative Real-time PCR whereas Pgp by Western blot, Rhodamine123 (Rho123) efflux ratio by Flow Cytometry. Results AdS-EGFP-shRNA1-U6 was succesfully constucted with high virus titer of 6×1010 pfu/ml. The interference efficency of AdS-EGFP-shRNA1-U6 agianst mdrlb in rat astrecyte model was about 94%. The Rho123 efllux ratio was about 15. 8%, significiently lower than control group which was 56. 2% (F = 127.5, P < 0. 05). Conclusions Pgp over-expression has been successfully suppressed and MDR has been reversed, which may provide a premising approach for refractory epilepsy remedy.

Objective To quantitatively study right ventricular function in the normal second and third trimester fetuses by M-mode tricuspid annular displacement(TAD).Methods TAD was measured using conventional M-mode echocardiography on 161 normal second and third trimester fetuses with gestation age (GA) between 19-38 weeks,meanwhile multiple parameters for evaluating right ventricular function were obtained using pulsed Doppler echocardiography (PW) and myocardial Dopple tissue imaging (DTI).The correlation between TAD and other parameters were analyzed using SPSS 17.0.Results In normal second and third trimester fetuses,the TAD was (9.38 ± 1.71)mm with a range of 5.79-13.90 mm,and was increased with the growth of GA.TAD was correlated with GA,E,A,Em,Am and Sm significantly (r =0.759,0.547,0.320,0.497,0.483 and 0.598 respectively,all P ＜0.001).TAD was not correlated with HR(P ＞0.05).TAD showed differences between the second trimester fetuses and the third trimester fetuses (P ＜ 0.05).Conclusions In normal second and third trimester fetuses,the TAD is increased with the growth of GA,and has good correlation with GA,E,A,Em,Am and Sm respectively,and may become a new promising modality to evaluate function of RV simply and accurately.The technique will be propitious to use in hospitals (without DTI) because of simplicity of operator and lower requirement on the technology and equipment precision.

AIM:To observe the effect of rapamycin (Rapa) on human neuroblastoma SH-SY5Y cell injury induced by oxygen-glucose deprivation (OGD), and to explore the role of autophagy in this process .METHODS: The SH-SY5Y cells were randomly divided into 4 groups:normal control group:the cells were cultured without OGD treatment;Rapa group:the cells were pretreated with Rapa for 1 h;OGD group:the culture medium was replaced by glucose-free me-dium and the cells were transferred to a humidified incubation chamber flushed by a gas mixture of 1%O2 , 94%N2 and 5% CO2 for 12 h; Rapa+OGD group: the cultured cells were treated with Rapa for 1 h, and then were given the same treatments as those in OGD group .The cell viability was assessed by MTT assay .The degree of the cell damage was evalu-ated by determining the leakage of lactate dehydrogenase ( LDH) .The enzyme activity of caspase-3 was detected .TUNEL staining were used to detect the variation of cell apoptosis .The protein levels of apoptosis-related proteins Bax and Bcl-2, autophagy-related protein beclin-1 and autophagy marker protein LC 3B were determined by Western blot .RESULTS:Compared with OGD group, the viability of the SH-SY5Y cells was significantly increased, and the activity of caspase-3 was significantly reduced in Rapa +OGD group (P<0.05).The SH-SY5Y cell injury was apparent after OGD with a great in-crease in the apoptotic rate (P<0.05).Compared with OGD group, the apoptotic rate significantly decreased in Rapa +OGD group (P<0.05).Compared with control group, the protein level of Bcl-2 was significantly decreased (P<0.05) and the protein level of Bax was significantly increased in OGD group .Compared with OGD group , the levels of Bcl-2, be-clin-1 and LC3B-Ⅱwere significantly increased and the protein level of Bax was significantly increased in Rapa +OGD group (P<0.05).CONCLUSION: Rapamycin has a protective effect on in vitro cultured SH-SY5Y cells injured by OGD.The mechanism may be related to the promotion of autophagy .

Objective To investigate the health-related quality of life and its relative factors in children with leukemia during maintenance treatment period. Therefore, the basis for effective individualization intervention can be provided. Methods A total of 224 leukemia children in maintenance treatment were collected in Affiliated Children′s Hospital of Chongqing Medical University from October 2015 to January 2016 by convenience sampling methods, questionnaire was used to assess the physical status, emotional status, social status, school status of the children. Results Single factor and generalized linear regression analysis were used to conclude that patient′s age (F=5.841-36.343, all P<0.01), parenting pattern (F=4.006-4.288, all P<0.05), family economic status (F=3.277-15.865, P<0.05 or 0.01) and the caregivers′information mastery level household location (F=2.044-2.661, P<0.05 or 0.01) had significant influence on the quality of life of children with leukemia in maintenance treatment. Conclusions The health-related quality of life and its relative factors in children with leukemia during maintenance treatment period should be analysis comprehensively. At the same time, take intervention should be taken from physiology, psychology, society, and school, to improve the social adaptability of children with leukemia and help them return to society as soon as possible.

BACKGROUND:Transpedicular transdiscal lumbar screw is a new type of spinal minimally invasive internal fixation technology.Compared with traditional bilateral pedicle screws,only one screw is needed to fix one segment on one side.It has the characteristics of being more economical,less trauma and easy to operate.However,studies on the application of transpedicular transdiscal lumbar screws combined with transforaminal lumbar interbody fusion(TLIF)and fixation are still rare. OBJECTIVE:To evaluate the effect of TLIF combined with various surgery methods on stress distribution of cage,fixation,disc lower and endplate and range of motion of lumbar vertebrae by constructing three kinds of finite element models including modified TLIF(cage alone)model,modified TLIF combined with bilateral pedicle screw(cage+BPS)model and modified TLIF combined with bilateral transpedicular transdiscal lumbar screw(cage+BTPTDS)model. METHODS:The CT images of the adult lumbar spine were used to establish the three kinds of TLIF finite element models:cage alone,cage+BPS and cage+BTPTDS using software Mimics,Geomagic and SolidWorks.ANSYS Workbench was used to simulate the application of six different motion loads of human body flexion and extension,left and right bending,and left and right rotation to calculate stress distribution and the changes in the range of motion of the lumbar spine of the cage,fixation,endplate and disc of the three lumbar spine surgery models and to compare the effects of three surgical options on the biomechanical effects of the lumbar spine. RESULTS AND CONCLUSION:(1)The cage alone model,cage+BPS model and cage+BTPTDS model were constructed successfully.(2)In flexion and lateral bending conditions,the maximum stress of the cage of cage+BTPTDS model was smaller than that of the cage alone model and a little greater than that of the cage+BPS model.In the extension condition,the maximum stress of the cage of the cage+BPS model was obviously smaller than that of the other two models.When it came to rotating condition,the maximum stress of the cage in the cage+BPS model and the cage+BTPTDS model presented no obvious difference,which was both smaller than the cage alone model.(3)The maximum stress of fixation of the cage+BTPTDS model was obviously bigger than the cage+BPS model in flexion and extension conditions,close to the cage+BPS model in lateral bending conditions,and smaller than the cage+BPS model in rotation conditions.(4)The maximum stress of the lower endplate of the fusion segment of the cage+BPS model was between the two other models.(5)In terms of the range of motion,the cage+BTPTDS model presented no obvious difference with that of the cage+BPS model at flexion and extension,left and right bending,and left and right rotation.(6)It is concluded that modified TLIF combined with transpedicular transdiscal lumbar screw provides stable support for the vertebral body of the fusion segment,ensures the motion range of the lumbar spine and has a good biomechanical effect.

OBJECTIVETo study the morphological variations of Paris polyphylla var. yunnansensis in different population for genetic diversity and breeding.

METHODThe characters of roots, stalks, leave and flowers were observed. The results were analyzed by DPS software.

RESULT AND CONCLUSIONP. polyphylla var. yunnansensis showed plenty genetic diversity, there existed obvious differences in morphological characters of different population. Principal components analysis showed that the number of calyces, petal, carpels, stamens is main factor,which causes the morphological variations in different population. Cluster analysis shows that 26 populations are incorporates in two types as 45.08 Euclidean distance. Leaf area index is distinct different in this two types.

Identification of the function of all genes in the mammalian genome is critical in understanding basic mechanisms of biology. However, the diploidy of mammalian somatic cells has greatly hindered efforts to elucidate the gene function in numerous biological processes by mutagenesis-based genetic approaches. Recently, mouse haploid embryonic stem (haES) cells have been successfully isolated from parthenogenetic and androgenetic embryos, providing an ideal tool for genetic analyses. In these studies, mouse haES cells have already shown that they could be used in cell-based forward or reverse genetic screenings and in generating gene-targeting via homologous recombination. In particular, haES cells from androgenetic embryos can be employed as novel, renewable form of fertilization agent for yielding live-born mice via injection into oocytes, thus showing the possibility that genetic analysis can be extended from cellular level to organism level.
Animals ; Embryonic Stem Cells ; cytology ; metabolism ; Genetic Techniques ; Genome ; Haploidy ; Models, Animal ; Mutagenesis ; Parthenogenesis

Objective:To investigate the in vitro inhibitory activity of a novel class Ⅰ and Ⅱb selective histone deacetylase (HDAC) inhibitor, purinostat mesylate (PM) , in diffuse large B-cell lymphoma and its mechanism.Methods:The 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide method was used to detect the effect of PM on cell proliferation. The effects of PM on cell cycle and apoptosis were detected by flow cytometry. The acetylation levels of HDAC substrate, cell cycle protein, apoptosis-related protein, and oncogene protein expression were detected by Western blot.Results:PM significantly inhibited the proliferation of lymphoma SUDHL-4 and SUDHL-6 cells and increased the acetylation levels of HDAC substrates H3, H4, and α-tubulin. In cell cycle experiments, PM induced G 0/G 1 phase arrest in SUDHL-4 and SUDHL-6 cells. Western blot experiment showed that PM could significantly downregulate the expression of cyclin-dependent kinases Cdk2, Cdk4, Cdk6, cyclin D1, and cyclin E and upregulate the expression of CDK inhibitor protein p21. In the apoptosis experiment, PM could induce the apoptosis of SUDHL-4 and SUDHL-6 cells. Western blot experiment demonstrated that PM promoted endogenous apoptosis by activating caspase-3 kinase and affecting antiapoptotic protein Bcl-2. In addition, PM could downregulate the expression of oncogene marker proteins MYC, IKZF1, and IKZF3. Conclusion:PM has an efficient biological activity in vitro for diffuse large B-cell lymphoma, including double-hit lymphoma, and provides valuable experimental evidence for PM in clinical treatment.

[Objective] To identify the ABO blood group of a patient with rare B antigen-specific autoantibody with cold agglutinin, and evaluate the effect of blood transfusion. [Methods] Red blood cells of patient were washed with 37℃ physiological saline and treated with sulfhydryl reagent. ABO blood group antigen was detected by tube method and microcolumn gel method. After the cold agglutinin was removed by EDTA anticoagulant plasma absorbed by type O red blood cells at 4℃, the related blood group antibodies were detected by type B red blood cells absorbing and releasing liquid at 4℃. The blood transfusion effect of patients was evaluated by the changes of hemoglobin before and after transfusion, and their ABO blood group was continuously monitored. [Results] B antigen was detected in the positive setting of serological experiment, cold agglutinin was detected by absorption and elution of type O red blood cells, and anti-B antibody was detected by absorption and elution of type B red blood cells. That is, there was specific autoantibody against B antigen, and the antibody property was IgM. No adverse reactions occurred during the infusion of 3 U type O washed red blood cells and the infusion was effective. The patient was continuously followed for two months, and the forward and reverse blood group identification were consistent, both of which were type B. [Conclusion] According to the previous blood group identification results, serological identification and follow-up comprehensive analysis, the ABO blood group of the patient is type B, but there are transient high titer cold agglutinin and B antigen-specific autoantibodies.

Objective To investigate the mechanism underlying the neuroprotective effect of linarin(LIN)against microglia activation-mediated inflammation and neuronal apoptosis following spinal cord injury(SCI).Methods Fifty C57BL/6J mice(8-10 weeks old)were randomized to receive sham operation,SCI and linarin treatment at 12.5,25,and 50 mg/kg following SCI(n=10).Locomotor function recovery of the SCI mice was assessed using the Basso Mouse Scale,inclined plane test,and footprint analysis,and spinal cord tissue damage and myelination were evaluated using HE and LFB staining.Nissl staining,immunofluorescence assay and Western blotting were used to observe surviving anterior horn motor neurons in injured spinal cord tissue.In cultured BV2 cells,the effects of linarin against lipopolysaccharide(LPS)-induced microglia activation,inflammatory factor release and signaling pathway changes were assessed with immunofluorescence staining,Western blotting,RT-qPCR,and ELISA.In a BV2 and HT22 cell co-culture system,Western blotting was performed to examine the effect of linarin against HT22 cell apoptosis mediated by LPS-induced microglia activation.Results Linarin treatment significantly improved locomotor function(P<0.05),reduced spinal cord damage area,increased spinal cord myelination,and increased the number of motor neurons in the anterior horn of the SCI mice(P<0.05).In both SCI mice and cultured BV2 cells,linarin effectively inhibited glial cell activation and suppressed the release of iNOS,COX-2,TNF-α,IL-6,and IL-1β,resulting also in reduced neuronal apoptosis in SCI mice(P<0.05).Western blotting suggested that linarin-induced microglial activation inhibition was mediated by inhibition of the TLR4/NF-κB signaling pathway.In the cell co-culture experiments,linarin treatment significantly decreased inflammation-mediated apoptosis of HT22 cells(P<0.05).Conclusion The neuroprotective effect of linarin is medicated by inhibition of microglia activation via suppressing the TLR4/NF-κB signaling pathway,which mitigates neural inflammation and reduce neuronal apoptosis to enhance motor function of the SCI mice.