Objective:This study aims to analyze the differences in the alternative splicing pattern of ADAR2 among glioma cell lines U87, U251, A172, and normal human astrocyte HA1800. Methods:A-to-I editing level at the Q/R-Site of GluR-2 was analyzed by RT-PCR and sequencing. Real-time PCR was performed to detect the expression level of each alternatively splicing variant using a specific primer that was confirmed to amplify only the targeted template and not other alternatively spliced variant fragments. Results:We verified that the Q/R-Site of GluR-2 is under-edited in glioma cell lines. Real-time PCR revealed that the ADAR2 pre-mRNA splic-ing pattern has no significant difference at exons 1a and 2 between glioma cell lines and normal human astrocyte. We also detected that the amount of alternative splicing variants, including exon 5a, was higher than that of alternative splicing variants not including exon 5a in human glioma cell lines. However, the expression of alternative splicing variants, including exon 5a, was lower than that of alterna-tive splicing variants not including exon 5a in human astrocyte. Conclusion:Evident differences in splicing were observed at the site of exon 5a between glioma cell lines and normal human astrocytes. The difference in the alternatively splicing pattern at exon 5a may be attributed to the decreased activity of ADAR2.

Objective To observe the effects of hydrogen sulfide (H2S) on structure and function of mitochondria of lung in rats with acute lung injury (ALI) induced by lipopolysaccharide (LPS).Methods Forty healthy male Sprague-Dawley (SD) rats were randomly divided into control group, LPS injury group, and low-, middle-, and high-dose NaHS groups, with 8 rats in each group. The rats in LPS injury group were given LPS 5 mg/kg via sublingual vein, and those in low-, middle-, and high-dose NaHS groups were challenged by LPS for 3 hours followed by intraperitoneally injection of 0.78, 1.56 and 3.12 mg/kg NaHS respectively in a volume of 2 mL/kg. The rats in control group were given 2 mL/kg normal saline via sublingual vein. The rats were sacrificed at 6 hours after model reproduction, and the lung tissues were harvested on time. The mitochondria in lung tissues were isolated with differential centrifugation. The lung mitochondria ultra structures were observed with electron microscope. The content ofmalondialdehyde (MDA) in mitochondria was determined with thiobarbituric acid method, and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and adenosine triphosphatase (ATPase) were determined with xanthine oxidase method. The mitochondrial activity and swelling were determined by multiskan spectrum.Results It was shown by transmission electron microscope that the mitochondrial structure in the control group was normal. The mitochondria in rat lung cells was swollen with disrupted or disintegrated cristae, the osmiophilic lamellar bodies had fused or disappeared, and rough endoplasmic reticulum degranulation phenomenon was obvious in LPS injury group. The mitochondrial damage was slightly mitigated in the low-dose NaHS group, and it was significantly mitigated in the middle-dose and high-dose NaHS groups. Compared with control group, the MDA content in lung mitochondria in LPS injury group was significantly increased (nmol/mg: 26.30±1.45 vs. 11.16±1.20), andSOD, GSH-Px, and ATPase activities were significantly decreased [SOD (U/mg): 18.78±1.13 vs. 27.44±1.97, GSH-Px (U/mg): 63.91±1.99 vs. 128.15±3.47, ATPase (U/mg): 4.83±0.25 vs. 9.92±0.65]; as well as the activity of the mitochondria was significantly decreased (A value: 0.164±0.025 vs. 0.319±0.045), and the swelling of the mitochondria was significantly increased (A value: 0.182±0.012 vs. 0.273±0.023), all with significantly statistical differences (allP < 0.01). Compared with LPS injury group, the MDA contents in low-, middle-, and high-dose NaHS groups were significantly decreased (nmol/mg: 21.89±1.23, 17.63±1.56, 12.19±1.30 vs. 26.30±1.45), and the SOD, GSH-PX, and ATPase activities were significantly increased [SOD (U/mg): 20.13±0.85, 21.38±1.22, 24.05±1.56 vs. 18.78±1.13; GSH-Px (U/mg): 82.06±1.65, 101.45±2.14, 117.80±2.12 vs. 63.91±1.99; ATPase (U/mg): 5.34±0.23, 7.06±0.37, 8.78±0.44 vs. 4.83±0.25]; as well as the activity of the mitochondria was markedly increased (A value: 0.194±0.018, 0.230±0.032, 0.297±0.038 vs. 0.164±0.025), and the swelling of mitochondria was markedly decreased (A value: 0.195±0.008, 0.219±0.017, 0.249±0.018 vs. 0.182±0.012), all with significantly statistical differences (allP < 0.05). Moreover, the protective effect of NaHS showed a dose-dependent manner.Conclusion It could be concluded that LPS induce mitochondrial structural damage and functional impairment in rats with ALI induced by LPS, and H2S have a beneficial effect against ALI induced by LPS with decreasing the mitochondrial lipid peroxidation level and protecting the cell structure and function, and the effect is correlated with the dosage.

Objective To investigate the effect of the synthetic rehabilitation treatment using the assistive devices for children with cerebral palsy.Methods 60 outpatients (male 31,female 29) diagnosed as cerebral palsy in our hospital from June 2009 to June 2010 were divided into trial group (n =30) and control group (n =30).Both of the two groups received our rehabilitation treatment three times a week.Compared with the daily family rehabilitation training of the control group,trial group still received a daily rehabilitation treatment by their parents using the assistive devices under the guidance of the therapist.They were assessed with Gross Motor Function Measure 66 items version (GMFM-66)and 88 items version (GMFM-88) before and after 6 months treatment.Results The total score of GMFM-66 and each dimension score of GMFM-88 improved in both group after 6 months treatment,except the centrol group Lying & Rolling dimension.Trial group 51 ± 13 vs.44 ± 15,control group 42 ± 17 vs.38 ± 16 (t =-3.580,P =0.001).There was significant difference on the total score of GMFM-66 and the three dimensions score (Lying & Rolling,Sitting and Walking,Running & Jumping) of GMFM-88 between two group.Lying & Rolling:F =4.59,P =0.036; Sitting:F =6.40,P =0.014 ; Walking,Running & Jumping:F =4.35,P =0.041.Conclusion The synthetic rehabilitation treatment in Medical institutions associate with family rehabilitation treatment using the assistive devices can effectively improve the recovery of gross motor function in children with cerebral palsy.

OBJECTIVETo investigate the anti-hyperlipidemic effects of apple polyphenols extract (APE) in Triton WR-1339-induced endogenous hyperlipidemic model.

METHODSFirstly, APE was isolated and purified from the pomace of Red Fuji Apple and contents of individual polyphenols in APE were determined using high-performance liquid chromatography-mass spectrometry (HPLC-MS). Secondly, forty male National Institude of Health (NIH) mice were randomly divided into 5 groups with 8 animals in each group. The Fenofibrate Capsules (FC) group and APE groups received oral administration of respective drugs for 7 consecutive days. All mice except those in the normal group were intravenously injected through tail vein with Triton WR-1339 on the 6th day. Serum and livers from all the mice were obtained 18 h after the injection. The changes in serum total cholesterol (TC), triglyceride (TG), lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) were measured by respective kits. Finally, expression of hepatic peroxisome proliferator-activated receptor alpha (PPARα) mRNA was measured by real-time reverse transcription-polymerase chain reaction (RT-PCR) method. RESULTS SERUM TC AND TG LEVELS SIGNIFICANTLY INCREASED IN TRITON WR-1339-INDUCED MODEL GROUP COMPARED WITH THE NORMAL GROUP (P<0.01). ORAL ADMINISTRATION OF APE [200 AND 400 MG/(KG DAY)] DOSE-DEPENDENTLY REDUCED THE SERUM LEVEL OF TG IN HYPERLIPIDEMIC MICE (P<0.01). SERUM LPL AND HTGL ACTIVITIES SIGNIFICANTLY DECREASED IN TRITON WR-1339-INDUCED MODEL GROUP COMPARED WITH THE NORMAL GROUP (P<0.05). ORAL ADMINISTRATION OF APE [200 AND 400 MG/(KG DAY)] DOSE-DEPENDENTLY ELEVATED THE SERUM ACTIVITY OF LPL IN HYPERLIPIDEMIC MICE (P<0.05 OR P<0.01). FURTHERMORE, COMPARED WITH THE NORMAL GROUP, HEPATIC MRNA LEVEL OF PPARα IN THE MODEL GROUP SIGNIFICANTLY DECREASED (P<0.01). ORAL ADMINISTRATION OF APE [200 AND 400 MG/(KG DAY)] DOSE-DEPENDENTLY ELEVATED THE EXPRESSION OF PPARα IN HYPERLIPIDEMIC MICE (P<0.05 OR P<0.01):

CONCLUSIONAPE could reduce TG level via up-regulation of LPL activity, which provides new evidence to elucidate the anti-hyperlipidemic effects of APE.

Animals ; Chlorogenic Acid ; pharmacology ; therapeutic use ; Cholesterol ; blood ; Flavonoids ; pharmacology ; therapeutic use ; Hyperlipidemias ; blood ; drug therapy ; enzymology ; pathology ; Hypolipidemic Agents ; pharmacology ; Lipoprotein Lipase ; blood ; genetics ; Male ; Mice ; PPAR alpha ; genetics ; metabolism ; Phytotherapy ; Polyethylene Glycols ; RNA, Messenger ; genetics ; metabolism ; Tannins ; pharmacology ; therapeutic use ; Triglycerides ; blood ; Up-Regulation ; drug effects

OBJECTIVE To improve uptake and cytotoxicity of the drug on MCF-7 cells by developing a poly (N-isopropylacrylamide) (PNIPAm) nanogel for Quercetin (Que) METHODS The PNIPAm nanogel was optimized by an orthogonal design and its structure was confirmed by FT-IR.A single factor experiment was used to optimize the formulation of quercetinloaded nanogel (Que-PNIPAm).The particle size,surface morphology and drug loading were characterized and the in vitro release behavior was investigated.Cytotoxicity of MCF-7 cells induced by Que-PNIPAm was investigated by CCK-8 method.The qualitative and quantitative cellular uptake studies were investigated by fluorescence microscope and flow cytometry,respectively.The mechanism of cellular uptake was investigated by the inhibitor method.RESULTS The particle size and drug loading of Que-PNIPAm were measured as (166.1±2.87)nm and 3.18％,respectively.Nanogel exhibited spherical morphology and uniform size distribution observed by electron microscopy.Compared to free Que,Que-PNIPAm significantly increased inhibition rate of MCF-7 cells.Que-PNIPAm also showed higher cell uptake efficiency and more effective antitumor activity at 42 ℃.Colchicine and 2-deoxyglucose have an inhibitory effect on MCF-7 cells uptake.CONCLUSION The prepared nanogel shows small particle size,thermosensitive property,which could significantly enhance the capacity of cellular uptake and tumor cytotoxicity.The mechanism of cellular uptake demonstrates tubulin is involved in the internalization of the nanogel into MCF-7 cells.

OBJECTIVETo explore the intrinsic connection between activation of classical nuclear factor-κB (NF-κB) pathway and gefitinib resistance in human lung adenocarcinoma H1650 cells.

METHODSHuman lung adenocarcinoma H1650 cells were exposed to gefitinib continuously for 60 days to obtain resistant H1650 cells. The expressions of P-IκBα, P-p50 and P-p65 in the cytoplasm or nuclei were detected using Western blotting in human lung adenocarcinoma HCC827 cells, parental H1650 cells and gefitinib-resistant H1650 cells. The effects of gefitinib alone or in combination with PDTC on the survival rate and expressions of NF-κB P-p50 and P-p65 were compared among the 3 cell lines.

RESULTSGefitinib-resistant H1650 cells showed increased cytoplasmic and nuclear P-IκBα expressions. The expressions of P-p50 and P-p65 differed significantly among the 3 cell line, decreasing in the order of resistant H1650 cells, parental H1650 cells, and gefitinib sensitive HCC827 cell lines (P<0.05 or 0.01). Treatment with gefitinib alone resulted in a significantly lower cell inhibition rate in resistant H1650 cells than in the parental H1650 cells (P<0.05) and HCC827 cells (P<0.01). The resistant H1650 cells had a significantly higher expression of P-p50 and P-p65 than other two cell lines (P<0.05). In both the resistant and parental H1650 cells, gefitinib significantly lowered P-p50 and P-p65 expressions (P<0.05 or 0.01), and the combined treatment with gefitinib and PDTC significantly decreased the cell survival rate and further lowered the cytoplasmic and nuclear expressions of P-p50 and P-p65 (P<0.01 or 0.01).

CONCLUSIONThe activation of classical NF-κB pathway is a key factor contributing to transformation of the parental H1650 cells into gefitinib-resistant cells. Gefitinib combined with PDTC can inhibit P-IκBα production and NF-κB P-p50 and P-p65 activation to suppress the survival of residual H1650 cells and the generation of gefitinib-resistant cells.

Objective :To explore rehabilitative effect of low—frequency (1Hz ,LF) repetitive transcranial magnetic stimulation (rTMS) combined functional electrical stimulation (FES) on lower limb spasm and motor function in pa—tients with subacute ischemic stroke (SAIS).Methods : A total of 92 SAIS patients with lower limb spasm and motor dysfunction hospitalized in our department were randomly divided into sham stimulation + FES group (sham stimu—lation group ,n＝45) and low—frequency rTMS + FES group (LF—rTMS group ,n＝47) ,both groups were continu—ously treated for three weeks .Lower limb motor function and spasm severity were assessed by Fugl—Meyer motor as—sessment (FMA) and modified Ashworth scale (MAS) respectively before and after treatment ,and motor evoked potential (MEP) was detected .Results : Compared with before treatment ,there was significant rise in FMA score , and significant reductions in MAS score and MEP in two groups after three—week treatment , P＝ 0.001 all ;com—pared with sham stimulation group ,there was significant rise in FMA score [16. 0 (13.0 ,23. 5 ) scores vs. 19. 0 (16.0 ,27.0) scores] ,and significant reductions in MAS score [1. 5 (1. 3 ,1.5) scores vs. 1. 0 (0. 5 ,1. 5) scores] and MEP [ (24.64 ± 0. 39) ms vs. (24.43 ± 0.31) ms] in LF—rTMS group after treatment , P＜0.05 or ＜ 0. 01 . Conclusion : Compared with sham stimulation + FES ,LF—rTMS + FES can more significantly improve lower limb spasm and motor function in patients with subacute ischemic stroke .

Background: Porcine circovirus type 2 (PCV2) infection is ubiquitous around the world.Diagnosis of the porcine circovirus-associated disease requires clinic-pathological elements together with the quantification of viral loads. Furthermore, given pig farms in regions lacking access to sufficient laboratory equipment, developing diagnostic devices with high accuracy, accessibility, and affordability is a necessity. Objectives: This study aims to investigate two newly developed diagnostic tools that may satisfy these criteria. Methods: We collected 250 specimens, including 170 PCV2-positive and 80 PCV2-negative samples. The standard diagnosis and cycle threshold (Ct) values were determined by quantitative polymerase chain reaction (qPCR). Then, two point-of-care (POC) diagnostic platforms, convective polymerase chain reaction (cPCR, qualitative assay: positive or negative results are shown) and EZtargex (quantitative assay: Ct values are shown), were examined and analyzed. Results: The sensitivity and specificity of cPCR were 88.23% and 100%, respectively; the sensitivity and specificity of EZtargex were 87.65% and 100%, respectively. These assays also showed excellent concordance compared with the qPCR assay (κ = 0.828 for cPCR and κ = 0.820 for EZtargex). The statistical analysis showed a great diagnostic power of the EZtargex assay to discriminate between samples with different levels of positivity. Conclusions The two point-of-care diagnostic platforms are accurate, rapid, convenient and require little training for PCV2 diagnosis. These POC platforms can discriminate viral loads to predict the clinical status of the animals. The current study provided evidence that these diagnostics were applicable with high sensitivity and specificity in the diagnosis of PCV2 infection in the field.

This article reported a case with primary cystic echinococcosis in the left femur.

This paper reports a case with alveolar echinococcosis metastatic to the brain. This case underwent 10-year antiechinococcosis treatment and operations of the liver, right kidney and brain. Following multiple operations and oral administration of albendazole, the case still had recurrence and distant metastasis of hepatic echinococcosis. It is suggested that early prevention, early diagnosis, early regular treatment and surgical radical treatment are critical to the treatment of alveolar echinococcosis.