Child, Preschool ; Heart Septal Defects, Ventricular ; complications ; Humans ; Leriche Syndrome ; complications ; Male

Objective:To investigate the arginine (Arg) sites in splicing factor 2/alternative splicing factor (SF2/ASF) methylated by protein arginine methyltransferase 1 (PRMT1). Methods:Wild-type and Arg93,Arg97,Arg109 mutant SF2/ASF plasmids were constructed,and GST-PRMT1,GST-SF2/ASF and arginine mutant GST-SF2/ASF fusion proteins were induced and purified. Methylation activity of PRMT1 on wild-type or mutant SF2/ASF protein and methylated sites of SF2/ASF were examined by methylation assay. The effect of SF2/ASF methylation on its subcellular localization was analyzed by immunofluorescence assay.Results:PRMT1 induced methylation of SF2/ASF at arginine,and PRMT1 did not methylate SF2/ASF when SF2/ASF was mutant at Arg93,Arg97 or Arg109,with Arg97 mutation showing the most profound inhibitory effect. Methylation of SF2/ASF did not affect its subcellular localization.Conclusion:SF2/ASF is a newly identified substrate of PRMT1; Arg93,Arg97 and Arg 109 are the three methylation sites in SF2/ASF,and Arg97 is the main methylation site. Methylation of SF2/ASF does not affect its subcellular localization.

OBJECTIVETo observe the effect of alcohol extracts from Pharbitidis Semen on the proliferation and metastasis of Lewis lung cancer, and study its anti-tumor mechanism.

METHODIn vitro, MTT assay and scratch assay were adopted to detect the effect of alcohol extracts from Pharbitidis Semen on the proliferation and metastasis of Lewis lung cancer cells. The cell autophagy was detected by the acridine orange staining. The gap-junction intercellular communication (GJIC) was investigated by the fluorescent yellow transfer. The expression of aquaporin 1 (AQP1) was analyzed by the Western blotting. In vivo, the subcutaneous implant model and the experimental pulmonary metastasis model of Lewis lung cancer in mice were established to evaluate the anti-tumor and anti-metastasis effects of alcohol extract from Pharbitidis Semen. The serum carcinoembryonic antigen (CEA) and beta2 microglobulin (beta2-MG) of mice bearing Lewis lung cancer were detected by the electrochemiluminesence immunoassay. The expressions of lung AQP1 and Connexin 43 (Cx43) were examined by the immunohistochemical method.

RESULTIn vitro, alcohol extracts from Pharbitidis Semen inhibited the cell proliferation in a dose-dependent matter, significantly prevented the cell migration, down-regulated AQP1 proteins of cells, promoted GJIC, and decreased the serum-free autophagy of tumor cells. In vivo, compared with untreated model mice, alcohol extracts from Pharbitidis Semen inhibited the tumor growth in a dose-dependent matter, prevented the tumor metastasis and prolonged the life span of mice bearing Lewis lung cancer, while decreasing serum CEA and beta2-MG of mice bearing Lewis lung cancer, enhancing the immumohistochemical staining intensity of Cx43 and weakening aquaporins AQP1 positive intensity.

CONCLUSIONAlcohol extracts from Pharbitidis Semen could prevent the proliferation and metastasis in Lewis lung cancer cells. Its mechanism may be related to the promotion of GJIC and the down-regulation of AQP1.

Animals ; Antineoplastic Agents ; administration & dosage ; Aquaporin 1 ; genetics ; metabolism ; Carcinoma, Lewis Lung ; drug therapy ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Connexin 43 ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Ipomoea ; chemistry ; Lung Neoplasms ; drug therapy ; genetics ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Metastasis ; Seeds ; chemistry

Objective To investigate the biological function of M122 in pathogenesis of MCMV in developmental brain disorders and brain damage, screening for mouse brain cDNA library interacting with M122 was performed by a yeast two-hybrid system. Methods The reconstructed bait plasmid pGBKT7-M122 was transformed into yeast cells AH109 and screened on the nutrient deficiency medium SD/-Trp. After express of the bait protein in AH109 yeast strains was detected by Western blot analysis, yeast-two hybrid screening was performed by mating AH109 with Y187 containing mouse brain cDNA library plasmid. The diploid yeast cells were plated on the nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade. The second screening was performed with SD/-Trp/-Leu/-His/-Ade containing X-α-gal. The plasmids in positive colonies were extracted and transformed into E. coli JM109 cells. After plasmid DNA in JM109 cells were extracted form positive colonies and sequenced, the results were analyzed by bioinformatic methods. The interactions between M122 protein and the protein obtained from positive colonies were further confirmed by repeating yeast-two hybrid. Then, autoactivations of the proteins obtained from positive colonies were detected.Results The reconstructed bait plasmid was transformed into yeast cells AH109 successfully. The bait protein expressed in the yeast cells AH109 stably. 24 proteins interacting with MCMV M122 were screened, including syntaxin 8 ( Stx8 ), phosphoglucomutase 2 ( Pgm2 ), potassium voltage-gated channel, shaker-related subfamily, beta member 1 ( Kcnab1 ), collagen, type ⅪⅩ, alpha 1 ( Col19a1 ), archain 1 ( Arcn1 ), cytidylate kinase( Cmpk), DnaJ(Hsp40) homolog, subfamily A, member 1 (Dnaja1), ATPase, Na+/K + transporting, beta 3 polypeptide( Atp1b3 ), SH3-domain GRB2-like ( endophilin ) interacting protein 1 ( Sgip1 ),ankyrin repeat domain 17 (Ankrd17), Smg-7 homolog, nonsense mediated mRNA decay factor(Smg7),sperm associated antigen 9 ( Spag9 ), FK506 binding protein 1a ( Fkbp1a), MYST histone acetyltransferase monocytic leukemia 4 ( Myst4), hyaluronan and proteoglycan link protein 1 ( Hapln1), autophagy-related 3 (Atg3), splicing factor, arginine/serine-rich 5 ( Sfrs5 ), zinc finger, C3HC-type containing 1 ( Zc3hc1 ),thioredoxin-related transmembrane protein 1 ( Txndc1 ), adaptor protein complex AP-1, gamma 1 subunit (Ap1g1), Cullin 1 ( Cul1 ), and so on. Three of them were formerly unknown proteins. M122 protein could interact with the proteins obtained from positive colonies in the yeast cells AH109. Ap1g1 and Cul1 were proved to have autoactivation. Conclusion A class of proteins in brain interacting with M122 has been obtained. It is presumed that these proteins are correlated with neuropathogenesis of the brain disorders caused by CMV, but the candidates still need further confirmation for the interaction.

Objective To analyze the characteristics of toxin, the PCR-ribotyping(RT) and the multilocus sequence typing(MLST) of Clostridium difficile strains isolated from China-Japan Friendship Hospital in order to provide a basis for monitoring the outbreak of nosocomial Clostridium difficile infection.Methods A total of 321 samples were collected from the patients with suspected Clostridium difficile infection(CDI) in China-Japan Friendship Hospital(CJFH) during 2012 to 2013.All Clostridium difficile strains were isolated and identified by the standard phenotypic culture method.Cytotoxicity test was performed to detect toxin B.Toxin genes (tcdA and tcdB) and binary toxin genes (cdtA and cdtB) harbored by those strains were analyzed.RT and MLST were used for homologous analysis.Clinical data of the patients were collected to analyze the isolation rate of Clostridium difficile in different populations.Results Forty-eight strains of Clostridium difficile were isolated from 46 patients with diarrhea and three of them were isolated from the same patient.The incidence of CDI among all patients, outpatients and inpatients were 14.3%(46/321), 12.8%(5/39) and 14.5%(41/282), respectively.Toxin B was detected in all of the strains as indicated by the cytotoxicity test.Strains of sequence type 1(ST1) showed the strongest cytotoxicity of all the isolated Clostridium difficile strains.Ten out of the 48 strains (20.8%) were tcdA(-)/tcdB(+) strains, which belonged to either ST37 or ST81.The results of RT and MLST were consistent in assigning the strains into nine types, in which the predominant type was ST1/RT027 accounting for 27.1% (13/48).All of the ST1/RT027 strains presented a toxin gene profile of tcdA(+)/tcdB(+) and cdtA(+)/cdtB(+).Most of the ST1/RT027 strains were isolated from the Traditional Chinese Medicine Department of Respiratory, where smallnosocomial outbreaks of ST1/RT027 strain infection might happen.Conclusion CDI diagnosed in CJFH mainly belongs to nosocomial infection.Most of the isolated strains harbor tcdA(+)/tcdB(+) genes.Surveillance for the outbreaks of CDI caused by ST1/RT027 strains over producing toxins A and B should be strengthened in hospitals.

Objective To investigate the efficacy of initial polytherapy in the patients with newly diagnosed infantile spasm by evaluating the ambulatory electronecephalogram (EEG) profiles and serum neuron specific enolase ( NSE ) level.Methods Data from 108 cases of newly diagnosed infantile spasm from Sep.2007 to Nov.2010 in the Department of Pediatrics,the First Hospital of Jilin University,were respectively analyzed for the EEG profiles and serum NSE level.The patients were treated with multiple drugs.Results ( 1 )After treatment,EEG showed normal shape in 56 cases ( 62.2％ ),sporadic sharp and slow waves without hypsarrhythmia in 32 cases( 35.6％ ),and hypsarrhythmia in 2 cases (2.2％).The overall effective rate was 97.8％.(2) Serum NSE in 35 seizure-free cases were examined at the time points of hospital admission,48 hours and one month after seizure cessation,respectively.NSE level was significantly decreased at 48 hours after seizure control than that at hospital admission ( t =6.54,P ＜ 0.05 ) ; NSE at one month after seizure free was further decreased compared with that at admission point ( t =9.29,P ＜ 0.05 ).Conclusion Polytherapy for infantile spasms can ameliorate the abnormal EEGs.NSE level is decreased after seizure cessation.Intelligence is improved 6 months after seizure free and polytherapy may improve the prognosis of the children with infantile spasms.

Objective To evaluate the long term clinical effect of porcelain laminate veneer esthetic restoration for fluorosis teeth. Methods One hundred and thirty-two fluorosis teeth of 11 patients were restored with porcelain laminate veneer for esthetic purpose and the clinical effects were evaluated with modified Rygescriteria at 1.0 week, 0.5-year, 1.0-year, 1.5-year, and 2.0-year recalled time. Results After 2.0 years application, none of the veneers debonded, "A" degree rate of marginal adaptation was more than 85%, "A" degree rate of cavosurface marginal discoloration was more than 90%, "A" degree rate of secondary caries was more than 99%, "A" degree rate of post-operative sensitivity was more than 75%, and "A" degree rate of color match was more than 70%, "B" degree rate of color match was 100%. Conclusions The research indicates that the porcelain laminate veneer is a good esthetic restoration method for fluorosis teeth and the long term clinical effect is stable.

Objective To evaluate the diagnostic value of 64 multidetector-row CT angiography for internal carotid artery(ICA)stenosis and the application in the follow-up of carotid endarterectomy and percutaneous transluminal stenting.Methods Forty transient ischemie attack(TIA)patients with interpretable CTA and DSA of the cervical carotid arteries were selected from May 2005 to December 2005. This yielded a total of 80 vessels.The CTA curved planar reformations(CPR)and DSA images referenced to the distal cervical internal carotid were graded by two senior neuroradiologists blindly,according to the North American Symptomatic Carotid Endarterectomy Trial(NASCET)guidelines.The paired-t test was used to verify the statistical significant difference between pre-operating and post-operating of carotid endarterectomy or percutaneous transluminal stenting in measuring the vascular diameter and area of cross section using CTA.Results When the 70% stenosis was used as the cut-off value,the seasitivity,specificity,negative predictive value,and the positive predicting value were 97%,95%,95%,and 98%,respectively.There was statistically significant difference in measuring the vascular diameter(P

Objective To determine if stimulation of cholinergic anti-inflammatory pathway mediated by vagus can protect liver against sepsis.Methods Male SD rats weighing 250-300 g were anesthetized with intraperitoneal methane 1g?kg~(-1).Left common carotid artery,was cannulated for MAP monitoring and blood sampling.Sepsis was produced by ligation of cecum which was punctured twice at an interspace of 3 mm with a 9G needle(CLP).Bilateral vagus nerves were isolated,ligated with 4-0 silk and cut(VGX).The distal end of the vagus nerve was stimulated with direct current(5V,2 ms,1 Hz)continuously for 20 min(STM).Forty animals were randomly divided into 4 groups(n=10 each):group Ⅰ sham operation;group Ⅱ CLP;group Ⅲ CLP + VGX and group Ⅳ CLP+VGX+STM.Arterial blood samples were obtained at 0,1,2 and 4 h after operation for determination of plasma TNF-? concentration and serum ALT and AST activities.The animals were then killed and the livers removed for ultrastructure examination with electron microscope.Results Electrical stimulation of the distal end of vagus nerve significantly attenuated the significant decrease in MAP and increase in plasma TNF-? concentration and serum AST and ALT activities and the damage to the organelle in the liver cell induced by sepsis.Conclusion Our results show that electrical stimulation of vagus nerve can protect liver from sepsis to some extent through cholinergic anti-inflammatory pathway.

50?mol/L induced markedly oncotic cell death,but no apoptosis was observed.TEM examination indicated that a form of cell death accompanied by cellular swelling,organelle swelling and vacuolization,mitochondrial swelling and cristae membrane loss,and nucleus swelling, chromatin scattering or karyolysis,which characterized as oncosis.When treated with 50,200?mol/L of ART for 5 h, the intracellular ROS level of Panc-1 cells markedly increased to 1.60 and 4.49 fold compared with that of untreated cells,respectively.Pretreatment with TCEP effectively attenuated ART-induced intracellular ROS level and decrease the oncosis in Panc-1 cells.Conclusions:ART exerts profound cytotoxic effects on Panc-1 cells and induces an oncosis- like cell death,which is quite different from apoptosis.The cellular generation of ROS and its peroxidation damage may be one of the mechanisms for its anti-tumor effect on pancreatic cancer.