There is increasing evidence that microcirculation hypoxia plays an important role in pathogenesis of inflammatory bowel disease (IBD).Hypoxia-inducible factors (HIFs)are transcriptional factors that serve as master regulators in ischemic and hypoxia injuries.Aims:To investigate the effect of HIF-2αon dextran sulfate sodium (DSS)-induced colitis in mice and its possible mechanism.Methods:Mx-Cre/LoxP recombination system was utilized to establish a conditional HIF-2αgene knockout (HIF-2α-/-)mouse model.C57BL/6,HIF-2α+/+and HIF-2α-/-mice were randomly allocated into DSS colitis group and water drinking group,respectively.Experimental colitis was induced by treatment with 4% DSS in drinking water for 7 days,and the disease activity index (DAI)was assessed daily.Mice in each group were sacrificed on day 1,3,5 and 7 in batch;the histopathological changes of colonic tissue were observed, and mRNA expressions of HIF-2αand tumor necrosis factor-α(TNF-α)were measured by real-time PCR.Results:During model establishment,expression of HIF-2αmRNA in colonic tissue was elevated in C57BL/6 and HIF-2α+/+DSS colitis groups,and the DAI and colonic inflammatory score were significantly higher than those in C57BL/6 water drinking group (P<0.05 on day 5 and day 7).Compared with HIF-2α+/+DSS colitis group,HIF-2α-/-DSS colitis group had more severe colonic inflammatory injury and the DAI and inflammatory score were further increased (P all<0.05,except the inflammatory score on day 7);expression of TNF-αmRNA in colonic tissue was also increased significantly in HIF-2α-/-DSS colitis group (P<0.05 on day 5 and day 7).Conclusions:HIF-2αmay ameliorate colonic inflammatory injury in mice with DSS colitis via inhibition of TNF-αexpression.

Image-guided neurosurgery is playing a more and more important role in modern neurosurgical therapy. The current trend is to construct an open platform for image-guided neurosurgery, to make applied researches of various new therapeutic techniques, and finally to integrate them into the clinical image-guided neurosurgery system.
Image Processing, Computer-Assisted ; Neurosurgery ; methods ; trends ; Surgery, Computer-Assisted ; trends

OBJECTIVETo detect angiotensin II (ANG II) in the maternal blood, umbilical cord blood, and at the maternal-fetal interface in women with pregnancy-induced hypertension (PIH) and explore the etiology of PIH and pathophysiologic mechanism of intrauterine growth restriction (IUGR).

METHODSEnzyme-linked immunosorbent assay (ELISA) was used to detect the concentration of ANG II in the maternal blood, cord blood, and maternal and fetal placental tissues in 30 women with PIH and 30 with normal pregnancy, and the results were analyzed with independent-sample t test and Pearson correlation analysis.

RESULTSANG II level in the maternal placental tissue homogenate showed no significant difference between women with PIH and normal pregnant women (8.51+/- 4.01 vs 7.76+/-3.47 pg/ml, P>0.05), but women with PIH had significantly higher ANG II in the fetal placental tissue (11.82+/-3.92 vs 9.64+/-2.63 pg/ml, P<0.05). ANG II level was significantly higher in the maternal blood of women with PIH than in normal pregnant women (46.44+/-8.48 vs 32.43+/-5.87 pg/ml, P<0.001), but similar in the cord blood (68.83+/-8.68 vs 72.47+/-8.51 pg/ml; P>0.05). A positive correlation was indicated between the cord blood and maternal peripheral blood ANG II levels in women with PIH (r=0.7379, P<0.05).

CONCLUSIONANG II in the fetal placental tissue is elevated, and the cord blood and maternal peripheral blood ANG II levels are positively correlated in women with PIH.

Angiotensin II ; blood ; isolation & purification ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Female ; Fetal Blood ; chemistry ; Fetal Growth Retardation ; blood ; Humans ; Hypertension, Pregnancy-Induced ; blood ; Placenta ; chemistry ; Pregnancy

The effects of beta-cypermethrin (consisting of alpha-cypermethrin and theta-cypermethrin) on the transient outward potassium current (I(A)) and delayed rectifier potassium current (I(K)) in freshly dissociated hippocampal CA3 neurons of rats were studied using whole-cell patch-clamp technique. The results indicated that alpha-cypermethrin increased the value of I(A) and theta-cypermethrin decreased the value of I(A), though both of them shifted steady activation curve of I(A) towards negative potential. theta-cypermethrin contributed to the inactivation of I(A). The results also showed that alpha-cypermethrin and theta-cypermethrin decreased the value of I(K), and shifted the steady state activation curve of I(K) towards negative potential. Both alpha-cypermethrin and theta-cypermethrin had no obvious effects on the inactivation of I(K). theta-cypermethrin prolonged recovery process of I(K). These results imply that both transient outward potassium channels and delayed rectified potassium channels are the targets of beta-cypermethrin, which may explain the mechanism of toxical effects of beta-cypermethrin on mammalian neurons.
Animals ; CA3 Region, Hippocampal ; cytology ; physiology ; Cells, Cultured ; Female ; Insecticides ; toxicity ; Male ; Neurons ; cytology ; drug effects ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Voltage-Gated ; drug effects ; physiology ; Pyrethrins ; toxicity ; Rats ; Rats, Wistar

The present study was designed to observe the influence of cerebral ischemia/reperfusion injury on learning and memory in hyperlipidemic rats and estimate the changes of activity of autonomic nervous system. Twenty-three male Wistar rats were randomly divided into three groups, named control group (C group, n=10), hyperlipidemia group (H group, n=6) and hyperlipidemia-ischemia group (HI group, n=7), respectively. The rats in H and HI group were fed a high-fat diet for 2 weeks and the rats in all groups were examined through Morris water maze (MWM) task. The rats in HI group underwent ischemia/reperfusion by 2-vessel occlusion (2-VO) method, and had electrocardiogram (ECG) recording simultaneously. The MWM task and ECG recording were taken again after 7 d of recuperation. The following results were obtained: (1) In the second place navigation performance and probe trial performance, the frequency of memory in quadrant of hidden-platform and memory score decreased significantly in HI group compared to that in C and H groups. (2) The heart rate in HI group decreased slowly after ischemia; the power at high frequency band (HF) reduced gradually, meanwhile the power at middle frequency band (MF) and the ratio of power at MF and HF decreased clearly compared to baseline value. (3) After 7 d of ischemia/reperfusion, the heart rate in HI group was significantly higher than that in H group (P<0.05). While there was no statistical change in the power at MF, the power at HF decreased and the ratio of MF/HF increased significantly (P<0.05). The data demonstrated that ischemia/reperfusion decreased the activity of autonomic nervous system, and the reduction of sympathetic nerve activity was much more than that of vagus nerve activity. The results suggest that the hippocampus neuron injury caused by ischemia induces cognitive disorder and imbalance of vago-sympathetic nerve activity accompanied by vagus nerve suppression.
Animals ; Autonomic Nervous System ; physiopathology ; Brain Ischemia ; etiology ; physiopathology ; Heart Rate ; physiology ; Hippocampus ; physiopathology ; Hyperlipidemias ; complications ; Learning Disorders ; etiology ; physiopathology ; Male ; Memory Disorders ; etiology ; physiopathology ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; pathology ; physiopathology

Objective To obtain shRNA sequences that can stably block the expression of Nuclear Factor kappa- B (p65) in the prostate cancer cell line LNCaP and construct the lentivirus vector.And validate the gene function of p65 in the cell line. Methods According to p65 genetic information, we design siRNA1, siRNA2, siRNA3 those three siRNA sequences targeting the ods area of p65 gene and then form the corresponding four pairs of complementary single strand DNA of shRNA, including the sense strand and the antisense strand. The synthetic shRNA sequence was inserted into the empty pSIH1-H1-copGFP shRNA Vector, and after transfecting the prostate cancer cells , the inhibitory effect of p65 mRNA by different sequences was detected through real-time PCR, and the inhibitory effect of p65 protein expression was detected by Western-blotting. Thus we can obtain highly effective shRNA sequences in the inhibition of p65 in prostate cancer cells. MTT, flow cytometry, transwell were chosen to test the cell growth, migration and invasive power in vitro to compare the difference of the experimental group, control group and negative group. Results The third shRNA sequence had the best inhibitory effect and the inhibitory effect of p65 mRNA in prostate cancer cell line was 59 % and the protein was 81%. It's position locates in p65 (NM_021975 ) 1096-1113 and it's stemloop sequence is 5'-GATCCGCCCTATCCCTTTACGTCATTCAAGAGATGACGTAAAGGGATAGGGCTTTTTG-3'. After transfecting, the prostate cancer cell line had the low expression of p65 stably. Through MTT, we got the growth curve, which showed that the growth ability of experimental group was significantly decreased compared with the control group and the Logarithmic growth didn't appear in the first 96 hours. Flow cytometry test displayed that the percentage of G0-G1-phase cells in experimental group was 61.49%, and the control group was 44.89%, idle group was 41.52%, which was increasing oberviously. The S-phase cells in the experimental group was 28.58%, compared with the 47.36% and 46. 10% diminished. The results of transwell showed that the experimental group had 16. 5000±6. 62076 cells and the other two groups had 45. 6333 13. 54159 and 36. 8333±5. 68412 cells, which showed the invasive power of experimental group was significantly declined(P＜0.05).Conclusions It's successful to obtain shRNA sequences that can stably block the expression of p65 in the prostate cancer cell line LNCaP and construct the lentivirus vector. p65 can positively regulates the biological behavior of prostate cancer LNCaP cell line in the cell growth, migration and invasive power.

OBJECTIVETo investigate the changes of gene expression file in transitional cell carcinoma of bladder after hepatocyte cell adhesion molecule(hepaCAM) overexpression.

METHODSAffymetrix Human Genome U133 Plus 2.0 Array was used to investigate the changes of gene expression profile between adenovirus-green fluorescent protein(GFP) -hepaCAM group and GFP group in transitional cell carcinoma of bladder EJ cells.Significant Analysis of Microarray(SAM) was used to screen the differentially expressed genes, DAVID software was used to conduct gene ontology analysis and wikiPathway analysis based on the differentially expressed genes. Reverse transcription-polymerase chain reaction and Western blot were applied to verify microarray data.

RESULTSCompared with the GFP group, a total of 2469 genes were up-regulated or down-regulated by more than 2 times in the GFP-hepaCAM group. Among these genes, 1602 genes were up-regulated and 867 were down-regulated.Most of the differentially expressed genes were involved in the function of cell proliferation and cell cycle regulation. The mRNA expressions of nibrin, liver kinase B1, and cyclin D1 detected by reverse transcription-polymerase chain reaction in three different bladder cancer cell lines were consistent with the microarray data.The protein expressions of nibrin and liver kinase B1 in these three cell lines measured by Western blot were consistent with the mRNA expression.

CONCLUSIONSHepaCAM can alter the gene expression profile of bladder cancer EJ cells. The well-known anti-tumor effect of hepaCAM may be mediated by regulating the gene expression via multiple pathways.

Carcinoma, Transitional Cell ; genetics ; pathology ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; physiology ; Humans ; Nuclear Proteins ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Proteins ; genetics ; physiology ; Urinary Bladder Neoplasms ; genetics ; pathology

Adolescent ; Adult ; Aged ; Body Fat Distribution ; Body Mass Index ; Diabetes Mellitus, Type 2 ; complications ; Fatty Liver ; etiology ; pathology ; prevention & control ; Female ; Humans ; Intra-Abdominal Fat ; metabolism ; pathology ; Male ; Middle Aged ; Obesity ; complications ; Risk Factors ; Severity of Illness Index ; Young Adult

Recently many reports have described that recombinant baculovirus could serve as a new gene transfer vehicle for mammalian cells with many unique advantages. In this study, the constructed recombinant baculovirus BacV-CMV-EGFPA containing the enhanced green fluorescent protein (eGFP) gene driven by CMV promoter was used to explore the feasibility of improving the efficiencies of transduction experiment in CV-1 cells by centrifugal method. Refer to the centrifugal transduction protocol of recombinant lentivirus, CV-1 cells were incubated with the culture supematant of Sf-21 cells infected by BacV-CMV-ECFPA (moi = 30) and then centrifuged at 600g for 1 h at RT, reporter gene transfer and expression efficiencies were analyzed by flow cytometry (FCM) 48h post transduction. Results showed that centrifugal method can achieve higher gene delivery and expression efficiencies than transduction by simple virus-cell mixing for 4h at 27 t with least impairment to cell viability. The centrifugal transduction protocol was further optimized by testing different centrifugal times, post-centrifugation incubation times and surrounding solutions. We found that centrifugation at 600g for 1 h at RT is sufficient to achieve the highest transduction efficiencies in target cells and PBS is more suitable than other surrounding solutions. Compared with previous protocol in which tranduction occurs for 4 - 8h at 27 degrees C, centrifugal method developed in this study could achieve more higher transduction efficiencies in more shorter time. Nine different mammalian cell lines (CV-1, 293FT, HepG2, 293T, CHO, C127, MT4, H9, Molt-4) were used to investigate the feasibility of delivering exogenous genes into different mammalian cells with the BacV-CMV-EGFPA supernatant (moi = 30) at 600g for 1h in PBS surrounding solution at RT. Results showed that most mammalian cell lines used in this study could be effectively transduced with recombinant baculoviruses by centrifugal method, and more higher and satisfactory transduction efficiencies could be achieved in primate adherent culture cells than in suspended culture cells. These results show that the baculovirus centrifugal transduction protocol have notable advantages: more rapid, efficient and nontoxic, and could be easily used in daily common experiments.
Animals ; Baculoviridae ; genetics ; CHO Cells ; Cell Line ; Cell Line, Tumor ; Centrifugation ; methods ; Cricetinae ; Cricetulus ; Feasibility Studies ; Flow Cytometry ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Hep G2 Cells ; Humans ; Microscopy, Fluorescence ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reproducibility of Results ; Spodoptera ; Transduction, Genetic ; methods

OBJECTIVETo find a better therapy for extensive cerebral infarction.

METHODSSixty cases with extensive cerebral infarction were treated by surgical decompression. After operation, they were randomly divided into an acupuncture plus medication group and a medication group, 30 cases in each group. The patients in the medication group were treated by routine western medicine, and the patients in the acupuncture plus medication group by the routine western medicine plus acupuncture 2 days after operation with Danzhong (CV 17), Zhongwan (CV 12), Qihai (CV 6), Zusanli (ST 36), Xuehai (SP 10) and Shuigou (GV 26) selected. They were treated for 10 days. The nervous functions were assessed by Chinese Stroke Nerve-function Rating Scale and criteria for clinical therapeutic effect.

RESULTSAfter surgical decompression, the death rate was 20.0%; after operation, acupuncture could significantly improve the neurological functional deficit degree of the patient. The total effective rate of 40.0% in the acupuncture plus medication group was better than 13.3% in the medication group (P<0.05).

CONCLUSIONThe surgical decompression is able to decrease the death rate of extensive cerebral infarction; after the operation, acupuncture and western medicine have cooperative effect on extensive cerebral infarction with a good therapeutic effect.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Cerebral Infarction ; drug therapy ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Nervous System ; physiopathology