OBJECTIVEThe aim of this study is to establish a transgenic mouse model for cleft palate relevant gene thyroid transcription factor-2 (TTF-2), which can be used to study palatal shelf development when the expression pattern and regular activation of TTF-2 is altered.

METHODSThe C57BL/6J mouse TTF-2 gene was cloned through polymerase chain reaction (PCR) from the mouse genomic DNA. The TTF-2 gene was inserted into the expression vector pBROAD3-mcs to construct the recombinant expression vector pBROAD3-TTF-2. This expression vector was then microinjected into the male pronuclei of the fertilized mouse ovum. Thus, the TTF-2 transgenic mice model was established. The genotype of the transgenic mice was identified by PCR and Southern blot analysis. Immunohistochemistry identified the consistent expression of TTF-2 gene during its palatal shelf development.

RESULTSTTF-2 genes were microinjected into 982 fertilized ova. A total of 580 two-cell-stage embryos cultured and transplanted into the oviducts of 48 pseudopregnant female mice. Overall, 68 embryos were obtained for analysis. The genotype of the mice was determined through PCR and Southern blot analysis using genomic DNA extracted from tail biopsies of the transgenic fetus. A total of 13 TTF-2 transgenic mice were detected. The expression of TTF-2 gene during the palatal shelf development of the transgenic mice was consistently detected by immunohistochemistry.

CONCLUSIONThe recombinant expression vector pBROAD3-TTF-2 was integrated into mouse genome through microinjection. The transgenic mouse in the palatal shelf that consistently expressed TTF-2 was successfully established and displayed a cleft palate phenotype.

Animals ; Cleft Palate ; Disease Models, Animal ; Female ; Forkhead Transcription Factors ; Genotype ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Thyroid Gland

Seventy-one patients with variant angina (VA) admitted in the Cardiology Department from January 2003 to March 2011,were divided into non-stenosis group (stenosis ＜ 50％,n =43) and stenosis group (stenosis ≥50％,n =28) according to the degree of stenosis.The differences of the risk factors,clinical manifestations,electrocardiogram,echocardiogram and laboratory examinations between these two groups were compared.The average age of patients in stenosis group 58 ± 8 y was higher than that in non-stenosis group (52 ± 9 y,t =2.43,P =0.02).Other risk factors,including male gender,smoking,hypertension,diabetes mellitus and lipid disorder did not show any differences between the two groups.Percentage of patients with angina pectoris lasting less than 5 min was higher in stenosis group (x2 =5.98,P =0.02),while percentage of effort angina,seeking medical consultation ≤ 6 months of onset and hemodynamic disorders showed no difference.Laboratory examinations had no differences.It is difficult to determine whether the VA patient has fixed coronary stenosis by analyzing the risk factors,clinical manifestations and laboratory examinations; to determine the fixed coronary stenosis coronary angiography is necessary.

Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.

Objective:To observe influence of preoperative sufentanil delayed preconditioning (SPC) on perioperative cogni‐tive function and myocardial protection effect in patients undergoing on -pump coronary artery bypass grafting (OP‐CABG) .Methods:A total of 60 patients undergoing selective OPCABG were randomly and equally divided into sufentanil group (SPC group) and routine treatment control group (control group) ,blood sample was collected before operation ,1h , 6h ,12h ,24h and 48h after aorta declamping to measure level of cardiac troponin I (cTnT );and all patients received neu‐ropsychological assessment before and 7d after operation .Results:Compared with control group ,there was significant re‐ductions in serum cTnI levels [1h:(23.66 ± 3.15) ng/ml vs .(14.96 ± 2.13) ng/ml] in sufentanil group from 1h after aor‐ta declamping to 48h after CABG ,P<0.05 all .The inotropic agent usage score on 12h ,24h and 48h after entering ICU of sufentanil group was significantly lower than that of control group [12h:(6.4 ± 0.6) scores vs .(8.4 ± 0.6) scores] , P<0.05 all .Incidence rate of postoperative cognitive dysfunction (POCD) in sufentanil group was significantly lower than that of control group (13.3% vs .30.0% ,P=0.041) .Conclusion:Preoperative sufentanil delayed preconditioning posses‐ses myocardial protection effect , and reduce incidence rate of cognitive dysfunction so also possesses certain brain protection in patients undergoing on-pump coronary artery bypass grafting .

Objective To preparation a new cationic liposome of which the positive component is a new cytofectin as a gene delivery vehicle for laboratory research of gene transfection and gene therapy in lung diseases. Methods The new cationic liposome was prepared by film. Electron microscopy and gel electrophoresis were employed to define the condition for transfection. The prepared cationic liposome was used in the transfection of a fluorescence plasmid to lung cancer cell line SPC and the rate of the transfection was evaluated. Results The prepared new cationic liposome was global microcapsule in size of 100 nm-1 ?m. The highest transfection rate of 60% was attained when the liposome was in a ratio of (6-8)∶1 with plasmid. Conclusion The new cationic liposome is prepared with cytofectin N 4-spermine cholesteryl carbamate as its positive component, and sound transfection activity is observed under a certain circumstance. It provides a basis for gene transfection and gene therapy.

Objective:To explore the biological activities of interferon-? on the fibroblasts from rat palatal scar.Methods:Fibroblasts were cultured from rat palatal scar.The cells of pasage 4-6 were suspended into culture medium at (2.5)?105 cells/ml.Then the cells were cultured as fibroblasts-populated collagen latice(FPCL) with the final cell density of 5?104/ml.The cultures were exposed to IFN-?(U/ml) at 0,40, 400 and 4 000 for 24 h respectively.The cell proliferation was studied by MTT assay and FPCL contraction was studied by diameter measuring.Results:The absorbance of the cells treated with IFN-?(U/ml) at 0,40,400 and 4 000 was 0.247?0.014,0.235?0.014,0.190?(0.024) and 0.184?0.021 respectively,the contraction idex(%) of FPCL treated with above concentrations of IFN-? was 88.53,64.47,46.00 and 23.63 respectively.Conclusion:IFN-? may inhibit the proliferation of fibroblasts and the contraction of FPCL.

Objective To document retrospectively long-term quality of life (QOF) and safety of living kidney donors.Methods A total of 219 living-related kidney donors which can be followed up had donated their kidneys between May 2004 and Sap.2011.The renal function,complications and QOF were estimaged.Results Donors included 104 men and 115 women with age from 19 to 66 years.Follow-up period was from 12 to 103 months.No cases died.The mean serum creatinine (Scr) was (84.0± 18.7) μmol/L and creatinine clearance (Ccr) was (1.23 ± 0.37) ml/s over 12 months postoperation.The average Ccr was lower in donors age over 50 years than in younger donors.The kidney function was still abnormal in 3 elder donors at end of the study.Thirty donors had hypertension including 5 newly cases.Microscopic hematuria was found in 4 cases.Hyperlipidemia developed in 3 cases.Mild anemia occurred in 2 cases.Femoral head necrosis occured in 1 case.Majority of 18.26％ donors (40 cases) reported weak healthy feeling (mild impact in 31 cases,moderate impact in 7 cases and severe impact in 2 cases).Thirty-five donors reported mild pain of incision (31 cases occasionally,and 4 cases frequently).Conclusion Living kidney donors have good long-term QOF and safety though there still exist risks of renal impact.Close follow-up is required especially in elderly donors.Compliance of donors needs to be further improved.

Objective To explore the property of brain functional networks and cognitive function changes in patients with frontal lobe low-grade gliomas (LGG).Methods 8 cases of suspected frontal lobe LGG patients were undergone with resting-fMRI scanning to analyze the small-world property of the LGG,meanwhile the LGG groups had Montreal (MoCA) cognitive score exam compared with the control group.Results The value of MoCA was 22.5 ± 1.5,21.8 ± 2.0,and 27.9 ± 2.1 respectively with statistical significance (P ＜ 0.05) in the LGG groups and the control groups.The LGG group cognitive score was significantly lower than that in the control group with statistical significance (P＜ 0.05).As to threshold,the two groups were consistent with the small world property.The LGG local efficiency was smaller than that of the controls,the postoperative small world properties (σ=2.49) were lower than that the pre-operative (σ =2.68),the largest brain function areas of preoperative information transmission were respectively the supramarginal gyrus,posterior cingulate,insula,and the postoperative being the precuneus,calcarine sulcus and superior frontal gyrus.The maximum cluster coefficient of the preoperative functional network were respectively the entorhinal cortex,transverse temporal gyrus and the calcarine sulcus,and postoperative were Wilson,transverse temporal gyri and occipital gyrus.Preoperative information transmission path was less than the postoperative,and the small world properties were positively correlated with MoCA.Conclusion LGG accompany by the changes of cognitive function,and with the small world network property preand post-operation.

Artemisia lactiflora is an important medicinal plant in China. The antitumor and antioxidant activities of the extracts of 54 endophytic fungi from the plant were screened via MTT assay and DPPH scavenging radical assay, respectively. The bioactive strains were identified based on similarity of 5.8S gene and internal transcribed spacer (ITS) sequences. The results showed that extracts from ten (18.5%) isolates exhibited antitumor activity, and which from two (3.7%) isolates exhibited antioxidant activity. The Alternaria sp. GYBH47 strain was simultaneously having antagonistic activity against HL-60 leukemia, MCF-7 breast and COLO205 colon cell lines, and Phomopsis sp. GYBH42 strain having cytotoxic and antioxidant activities. The results indicated that endophytic fungi from Artemisia lactiflora are potential resources to find valuable bioactive components.
Antineoplastic Agents ; chemistry ; pharmacology ; Artemisia ; microbiology ; Biphenyl Compounds ; metabolism ; Cell Line, Tumor ; Endophytes ; chemistry ; classification ; physiology ; Free Radical Scavengers ; chemistry ; pharmacology ; Fungi ; classification ; physiology ; Humans ; Picrates ; metabolism

Background The aspheric intraocular lenses(IOLs)can reduce ocular spherical aberration to some degree.However,the clinical effect depends more on the IOL proper alignment.It becomes more important to study the IOL position in eye,Objective This study was to analyze the position alteration of IOL after phacoemulsification combined with implantation of one-piece soft IOLs.Methods In this prospective control study,80 eyes of 40 patients with age-related cataract were enrolled.The phacoemulsification with IOL implantation was performed in all the eyes.Decentration and tilt of IOL in the nasal superior,superior temporal,inferior temporal and nasal inferior quadrants(the intersection point of the system optical axis and the IOL maximum cross plane were regarded as the ordinate origin)were measured by rotating Scheimpflug camera(Pentacam Oculus)in 3 months postoperatively under the mydriasis condition.Written informed consent was obtained from each subject prior to this trial.Results In the right eye group,the IOL decentered toward temporal in 26 eyes(65％)and infratemporal in 16 eyes(40％).IOLs tilted temporally in the horizontal plane in 37 eyes(92.5％)and tilted inferiorly in the vertical plane in 34 eyes(85.0％).In the left eye group,IOLs decentered temporally 33 eyes(82.5％)and 20 IOLs (50％)infratemporally,IOLs tilted temporally in the horizontal plane in 37 eye(92.5％)and 36 IOLs(90％)tilted inferiorly in the vertical plane.There was no statistical difference for the intercomparsion of horizontal/vertical decentration in various quadrant in the right eye(F =0.221,0.792,P＞0.05).The obvious elevated horizontal decentration was found in the supertemporal and infratemporal quadrants compared with supernasal quadrant in the left eyes but there was no significant difference in the vertical decentration among 3 quadrants(F=0.576,P＞0.05).Decentrations were positively correlated with the tilt in both horizontal and vertical plane(right eye horizontal plane:r=0.374,P=0.002;right eye vertical plane:r=0.402,P=0.001 ;left eye horizontal plane:r=0.377,P=0.002；left eye vertical plane:r=0.347,P=0.002).Conclusions The one-piece soft IOLs(Adapt AO)decenter toward temporal mostly in 3 months after surgery,especially infratemporally in the eye.And the optical axis of the IOL tilt toward infratemperol mostly in both right and left eyes.The decentration and tilt are consisted in the corresponding direction between the right and left eyes.The position of the IOLs showed mirror symmetry between right and left eyes.The IOLs decentration show the positively correlation to tilt whatever in horizontal and vertical plane.