Aortic Valve ; surgery ; Cardiac Catheterization ; Heart Valve Prosthesis Implantation ; methods ; Humans

Cardiac Catheterization ; standards ; Heart Defects, Congenital ; therapy ; Humans

Objective To explore the activated brain region of acute epilepsy in cat model induced by pentylenetetrazol(FFZ)with manganese enhanced-functional MR imaging(ME-fMRI),and evaluate the application of ME-fMRI on localization of the activated brain.Methods Forty cats were divided into 4 groups by random number table method as epileptic A and B groups as well as control A and B groups. Cats of epileptic groups were injected with PTZ(55 mg/kg)intramuscularly,and those of control groups were injected with the saline at same dose.The behavior change in the epileptic and control group A was observed and electroencephalogram(EEG)was also undertaken.Cats of epileptic and control group B were performed ME-fMRI,and the percentage of the enhanced signal intensity was then calculated.Results After injection with PTZ(55 mg/kg)intramuscularly,epileptic seizure was all evoked,and then EEG recording showed spike-wave and polyspike-wave complexes.The neocortex of cats of epileptic group B was diffusely phanero-enhanced on ME-fMRI.The percent enhancement of signal intensity in cortex of frontal lobe,parietal lobe and occipital lobe was(34.6?5.7)% and that in cortex of temporal lobe with(22.9? 6.5)%,whereas those of control group B with(14.9?4.5)% and(11.6?3.2)% respectively.And there was significant difference between the above different localization of the brain in the two groups (t=-10.43,-5.46 respectively,P

Objective To observe the expression of cardiotrophin-1(CT-1) in ischemia-reinfusion cardiac muscle of rats and the effect of neuregulin-1(NRG-1).Methods The model of ischemia-reinfusion cardiac muscle of rats were prepared,35 rats were randomly divided into 4 groups:model group(n=8),NRG-1 pretreatment group(n=9),pseudo-surgery group(n=8) and normal control group(n=10).The CT-1 mRNA in the observed cardiac muscle of all groups was measured by RT-PCR and the relative amount of CT-1 mRNA were calculated,and for statistical treatment.Results The CT-1 mRNA of model group was(63.96?9.34),and it was higher than that of pseudo-surgery group(36.16?5.43)and normal control group(36.84?4.64).The significant differences were found in 3 groups(F=47.37 P

OBJECTIVETo study the prevalence of chondromalacia patella among college students and the correlation with sports injury.

METHODS354 students from gymnastic department and 429 from nongymnastic department with knee joint pain were selected. 184 students from gymnastic department and 342 from nongymnastic department were checked randomly by a surgeon. 77 patients (37 males, 40 females) from gymnastic department and 119 patients (62 males, 57 females) from nongymnastic department were diagnosed as chondromalacia patellae. The amount of exercise and the occurrence of sports injury were investigated in each student. All data were analyzed with SPSS 10.0 statistical software.

RESULTSThe prevalence of chondromalacia patella was 20.1% in female students and 11.6% in male students from gymnastic department, and 5.61% in female students and 4.92% in male students from nongymnastic department. The amount of exercise and the occurrence of sports injury to the knee joint in students from gymnastic department were greater than those from nongymnastic department.

CONCLUSIONSIn both female and male students, the prevalence of chondromalacia patella is higher in gymnastic department than nongymnastic department. Sports injury is an important cause of chondromalacia patella.

Adolescent ; Adult ; Age Distribution ; Athletic Injuries ; epidemiology ; pathology ; Cartilage Diseases ; diagnosis ; epidemiology ; Case-Control Studies ; China ; epidemiology ; Comorbidity ; Cross-Sectional Studies ; Female ; Humans ; Injury Severity Score ; Knee Injuries ; epidemiology ; pathology ; Male ; Patella ; pathology ; Prevalence ; Probability ; Prognosis ; Reference Values ; Risk Factors ; Sex Distribution

Objective To explore the expression of HCK gene during the cardiomyocyte differentiation of mouse embryonic stem cells and analyze the role of HCK gene in maintenance of pluripotency of embryonic stem cells.Methods Mouse embryonic stem cells were cultured,then induced to differentiate into cardiomyocytes.Total RNAs were isolated from mouse embryonic stem cells in the differentiation days:0 day(D0),the second day(D2),the fourth day(D4),the sixth day(D6),the eighth day(D8),respectively.The levels of HCK mRNAs were assessed by the method of semi-quantitive reverse transcriptase-polymerase chain reaction(RT-PCR).In the meanwhile,Total proteins were also isolated from mouse embryonic stem cells in the differentiation D0,D2,D4,D6,D8,and the levels of HCK proteins were evaluated by Western-blot.Results HCK mRNAs could be detected in the mouse embryonic stem cells in D0 and D2,however,they were undetectable from D4 to D8.The expression of HCK mRNAs was rapidly down-regulated during cardiomyocyte differentiation of mouse embryonic stem cells.Expression of HCK proteins,which coincided with HCK mRNAs,down-regulated during differentiation and couldn't be detected in D4.Conclusions With the cardiomyocyte differentiation of mouse embryonic stem cells,the expression of HCK in the levels of mRNA and proteins are sharply down-regulated;HCK may play an important role in maintaining the pluripotency of embryonic stem cell.

Objective To knockout interferon alpha/beta receptor subunit 1(IFNAR1) gene in human colorectal adenocarcinoma cells Caco-2 using clustered regularly interspaced short palinmic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system to construct IFNAR1 knockout Caco-2 cell line.Methods The single guide RNA(sgRNA)sequence was designed to specifically recognize the exon region of IFNAR1 gene using CRISPR/Cas9 technology,and the LentiCRISPRv2-IFNAR1-sgRNA recombinant plasmid was constructed.Caco-2 cells were infected with the plasmid packaged by lentivirus and screened by puromycin resistance.The obtained monoclonal cell lines were cultured by limited dilution method,which were verified for the effect of IFNAR1 gene knockout by target gene sequencing and Western blot,and detected for the mRNA levels of CXC chemokine ligand 10(CXCL10)and interferon-stimulatd gene 20(ISG20)in IFNAR1knockout cells by adding exogenous IFNβ.Results Sequencing results of plasmid LentiCRISPRv2-IFNAR1-sgRNA showed that the insertion sites were all located at the sticky end of BsmBⅠenzyme digestion.Two IFNAR1 knockout monoclonal cell lines were obtained.The sequencing results showed that Caco-2-IFNAR1-KO1 had 5 bp deletion in the sixth exon of IFNAR1,and Caco-2-IFNAR1-KO2 had 18 bp deletion and 1 bp insertion in the seventh exon.Compared with wild-type Caco-2 cells,Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells showed no expression of IFNAR1 protein.Compared with no IFNβ stimulation,the mRNA levels of CXCL10 gene(t = 0.566 and 1.268 respectively,P>0.05)and ISG20 gene(t =1.522 and 1.733 respectively,P>0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells stimulated by 50 ng/mL IFNβ showed no significant increase.While compared with those of wild-type Caco-2 cells,the mRNA levels of CXCL10gene(t = 6.763 and 6.777 respectively,P<0.05)and ISG20 gene(t = 5.664 and 5.65 respectively,P<0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells decreased significantly under the stimulation of 50 ng/mL exogenous IFNβ.Conclusion Caco-2 cell line with IFNAR1 knockout was successfully constructed by using CRISPR/Cas9 technology,and the downstream molecules activated by IFNAR(interferon alpha/beta receptor)in this cell line were obviously inhibited,which provided a powerful tool for further exploration of the innate immune response and replication packaging mechanism of Caco-2 cells after virus infection.

Seven compounds were isolated from fermentation extract of cave-derived Metarhizium anisopliae NHC-M3-2 by silica gel, semi-preparative HPLC and other chromatographic methods. Their structures were elucidated by UV, IR, MS and NMR methods as 2,3-dehydroindigotide G (1), (-)-regiolone (2), naphtho-γ-pyrone (3), indigotide G (4), indigotide B (5) destruxin A (6) and destruxin B (7). Compound 1 is a new glycoside naphthopyranone compound. The anti-hepatitis B virus (HBV) activity of these compounds was evaluated. The EC₅₀ and CC₅₀ of compound 3 against HBV were 4.5 μmol·L^-1 and 92.3 μmol·L^-1, respectively. This is the first report of the antiviral activity of compound 3.

BACKGROUNDInfections caused by gram-negative bacteria (GNB) often lead to high mortality in common clinical settings. The effect of traditional antibiotic therapy is hindered by drug-resistant bacteria and unneutralizable endotoxin. Few effective methods can protect high risk patients from bacterial infection. This study explored the protection of adeno-associated virus 2 (AAV2)-bacteriacidal permeability increasing protein 700 (BPI(700))-fragment crystallizable gamma one 700 (Fc gamma1(700)) chimeric gene transferred mice against the minimal lethal dose (MLD) of E. coli and application of gene therapy for bacterial infection.

METHODSAfter AAV2-BPI(700)-Fc gamma1(700) virus transfection, dot blotting and Western blotting were used to detect the target gene products in Chinese hamster ovary-K1 cells (CHO-K1cells). Reverse transcription-polymerase chain reaction and immunohistochemical assay were carried out to show the target gene expression in mice. Modified BPI-enzyme linked immunosorbent assay was used to identify the target gene products in murine serum. The protection of BPI(700)-Fc gamma1(700) gene transferred mice was examined by survival rate after MLD E. coli challenge. Colony forming unit (CFU) count, limulus amebocyte lysate kit and cytokine kit were used to quantify the bacteria, the level of endotoxin, and proinflammatory cytokine.

RESULTSBPI(1-199)-Fc gamma1 protein was identified in the CHO-K1 cell culture supernatant, injected muscles and serum of the gene transferred mice. After MLD E. coli challenge, the survival rate of AAV2-BPI(700)-Fc gamma1(700) gene transferred mice (36.7%) was significantly higher than that of AAV2-enhanced green fluorescent protein (AAV2-EGFP) gene transferred mice (3.3%) and PBS control mice (5.6%). The survival rate of AAV2-BPI(700)-Fc gamma1(700) gene transferred mice treated with cefuroxime sodium was 65.0%. The bacterium number in main viscera, the levels of endotoxin and proinflammatory cytokine (tumor necrosis factor-alpha and interleukin-1beta) in serum of the AAV2-BPI(700)-Fc gamma1(700) gene transferred mice were markedly lower than that of PBS control mice (P < 0.01).

CONCLUSIONSAAV2-BPI(700)-Fc gamma1(700) gene transferred mice can resist MLD E. coli infection through expressing BPI(1-199)-Fc gamma1 protein. Our findings suggested that AAV2 mediated BPI(700)-Fc gamma1(700) gene delivery could be used for protection and treatment of clinical GNB infection in high-risk individuals.

Animals ; Anti-Bacterial Agents ; therapeutic use ; Antimicrobial Cationic Peptides ; Blood Proteins ; CHO Cells ; Cricetinae ; Dependovirus ; genetics ; Disease Models, Animal ; Escherichia coli Infections ; therapy ; Gene Transfer, Horizontal ; Genetic Therapy ; Mice ; Mice, Inbred BALB C ; Proteins ; genetics ; Receptors, IgG ; genetics ; Recombinant Fusion Proteins ; genetics

Objective To explore the expression of cardiotrophin-1(CT-1) in myocardium and peripheral blood plasma of neonatal rat with asphyxia and the regulative effect of neuregulin-1(NRG-1).Methods Ninety seven-day-old neonatal rats were randomly divided into 3 groups:asphyxia group (n=40),normal control group (n=10)and NRG-1 pretreatment group (n=40).The model of neonatal rat with asphyxia was prepared by the way of ligation of carotid combined with low supply of oxygen.NRG-1(1 mg/kg) was given to NRG-1 pretreatment group by intraperitoneal injection 30 min before asphyxia.The separated plasma of peripheral blood and myocardium antetheca of aortic ventricle of heart were taken at the time point of 6,12,24 and 48 h.The expression of CT-1 in peripheral blood plasma was detected by enzyme linked immunoadsorbent assay,and that of myocardium was determined by Western blot.Results The expressions of CT-1 protein in peripheral blood plasma of asphyxia group were significantly higher than those of normal control group at each time point,and reached the peak at 24 h(Pa