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

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.

10.3969/j.issn.1000-3606.2013.06.016

Objective To compare the effect of three types of lipid emulsions on the IL-1βand IL-6 expressions in acute lung injury induced by lipopolysaccharide (LPS) in rats. Methods One hundred young SD rats were divided into control,LPS,ω-6,ω-9 andω-3 group. Age 29 days, the control group were intravenously injected with saline,ω-6 group injected lipid emu-sion (C14-24),ω-9 group injected long chain fat emusion,ω-3 group injected fish oil fat emusion. Then the control group fol-lowed by intratracheal instillation of saline, and the reamining four groups were sprayed with LPS. Pathologic changes in lung tissue section were observed. The expression levels of IL-1βmRNA and IL-6 mRNA were determined by RT-PCR,and the con-centrations of IL-1βand IL-6 in bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immune-specific assay. Results Infiltration and bleeding were observed in lung tissue under light microscopy in ALI rats. The lung indexes and patho-logical scores of ALI model groups were significantly higher than those of control group (P<0.005). The expressions of IL-1βand IL-6 mRNA in groupsω-9 andω-3 were significantly higher than those in groups LPS andω-6 (P<0.005). The levels of IL-1βand IL-6 in BALF in groupsω-9 andω-3 were significantly higher than those in groups LPS andω-6 (P<0.005). There was no significant difference (P>0.005) of the expressions of IL-1βand IL-6 mRNA betweenω-9 andω-3 group. Conclusions ω-6 PUFAs can accentuate inflammation by up-regulating the levels of IL-1βand IL-6 whileω-9 PUFAs andω-3 PUFAs can relieve inflammation by down-regulating the levels of IL-1βand IL-6.

Objective To investigate the diagnostic value of carotid artery ultrasonography,CT angiography (CTA)and digital subtraction angiography (DSA)for carotid artery dissection. Methods The image data of carotid artery ultrasonography,CTA,and DSA of 24 patients with carotid artery dissection were analyzed retrospectively. Results Twenty-four,16,and 21 patients were examined with DSA,CTA,and carotid artery ultrasonography respectively. The detection rates of carotid artery dissection with DSA,CTA, and carotid artery ultrasonography were 95. 8%,75.0%,and 71. 4% respectively. The DSA mostly showed the line-like sign (n=12,50 %). CTA and carotid artery ultrasonography mostly showed the double lumen sign;they were 37. 5%(n=6)and 52. 4%(n=11)respectively. Compared with DSA,the concordance rates of carotid artery ultrasonography and CTA were 66. 7% and 81. 3% respectively. There was no significant difference (Kappa=0. 39,P=0. 08 and Kappa=0. 43,P =0. 22 respectively). The concordance rate of ultrasonography in combination with CTA and DSA reached 87. 5%(n=15,Kappa=0. 67,P =0.047). There was significant difference. Conclusion DSA is a gold standard for the diagnosis of carotid artery dissection,and it is irreplaceable. Carotid artery ultrasonography in combination with CTA can improve the diagnostic rate. Carotid artery ultrasonography can be used as a screening method for carotid artery dissection.

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 compare the efficacy and safety between Chinese generic imatinib (Xinwei(R),Jiansu Hansoh Pharmaceutical Group Co.,Ltd.) versus branded imatinib (Glivec(R),Novartis) in patients with newly-diagnosed chronic myeloid leukemia in chronic phase (CML-CP).Methods Patients with newly diagnosed CML-CP were enrolled and assigned to receive either Xinwei or Glivec at an initial dose of 400 mg/d according to patients' financial capability.The efficacy and adverse effects were evaluated.Results From January 2014 to September 2015,145 eligible patients were assigned to Xinwei (n =89) or Glivec (n =56) group.All patients were treated and followed up at least 3 months.At 3 months,the complete response rates were 95.5％ (85/89) and 100％ (56/56),major cytogenetic response rates were 74.2％ (66/89) and 80.4％ (45/56),and the proportions of achieving BCR-ALBIS ≤ 10％ were 76.1％ (67/88) and 82.1％ (46/56) in Xinwei and Glivec groups respectively (all P ＞0.05).With a median follow-up of 12 months,2 patients in each group progressed to accelerate or blast phase.Hematologic and non-hematologic side effects were similar between the 2 groups.Conclusions Early hematological,cytogenetic and molecular responses between Xinwei and Glivec are comparable in newly-diagnosed CML-CP patients.The progression rate and side effects are also similar between the 2 groups.

Objective To evaluate the predictive value of modified early warning score (MEWS) and SMART-COP score on mechanical ventilation in patients with severe influenza A H1N1. Methods Fifty cases diagnosed with severe influenza A H1N1 were retrospectively analyzed. The MEWS and SMART-COP score were calculated. The area under the receiver operating characteristic (ROC) curve (AUC) was evaluated using ROC curve. MEWS, SMART-COP score and AUC were analyzed by Z test. Results The AUCs of MEWS and SMART-COP score for predicting mechanical ventilation were 0. 923 and 0. 889, respectively, which were not significantly different (Z=0. 548, P =0. 584).Conclusion Both of MEWS and SMART-COP score are predictive factors of mechanical ventilation in the patients with severe influenza A H1N1.

Objective To investigate the nutritional status of vitamin D in preterm infants after birth and further explore its possible influencing factors, so as to guide clinical vitamin D therapy and to screen the preterm infants who are at high risk of vitamin D deficiency.Methods Retrospective analysis was conducted in the neonatal department of our hospital from April 21st, 2014 to February 5th, 2016.The serum 25(OH)D level in preterm infants were measured 2 weeks after birth.Data including gender, season of birth, time to initiation of breastfeeding were collected.According to the 25(OH)D levels[25(OH)D≤37.5 nmol/L, 37.5 nmol/L≤50.0 nmol/L, and 25(OH)D>50.0 nmol/L], all the preterm infants were divided into three groups: vitamin D deficiency, insufficiency, and sufficiency groups.The influencing factors of vitamin D in preterm infants were screened by using statistical method.Results The mean 25(OH)D level of 172 preterm infants was (43.1±16.7)nmol/L.In vitamin D deficient, insufficient, and sufficient groups, there were 68 (40％), 50 (29％) and 54(31％) cases of preterm babies, respectively.The mean values of 25(OH)D in these three groups were (27.8±16.7)nmol/L, (42.4±3.4)nmol/L, and (63.0±11.7)nmol/L, respectively.Only the season of birth had significant difference among three groups (P=0.013): 44.2％ of the preterm infants born in winter had vitamin D deficiency, which was higher than those in spring (41.7％), summer(33.3％), and autumn (38.1％);44.2％ of the preterm infants born in winter had vitamin D insufficiency, which was much higher than those in spring (30.6％), summer (25.1％), and autumn (19.0％);furthermore, only 11.6％ of the preterm infants born in the winter had vitamin D sufficiency, which was much lower than those in spring (27.8％), summer (41.2％), and autumn (42.9％) (OR=4.655, 95％ CI=1.716-12.627, P=0.003).Conclusion Vitamin D deficiency in preterm infants 2 weeks after birth is prevalent, and winter birth is a risk factor of vitamin D deficiency and insufficiency in preterm infants.

Objective To investigate the current situation of nursing staffs in second-class and tertiary hospitals of Nanchang City and influencing factors in order to provide a basis for the management and research of patients safety.Methods The convenience cluster sampling method was adopted to extract the whole nursing staffs from 7 hospitals(4 second-class hospitals and 3 tertiary hospitals)in Nanchang City as the research subjects.Then the multiple regression method was adopted to analyze the influencing factors of patients safety culture in the second-class and tertiary hospitals of Nanchang City.Results The advantage fields of cognition of the nursing staffs in the second-class hospitals on the patients safety culture were the intra-department teamwork,expectation actions of managers for promoting safety,organizational learning and continuous improvement,feedback and communication of mistakes,while the fields needing to be improved were the communication openness,personnel allocation,non-penalty reactions on mistakes;the advantage fields in the tertiary hospitals were same to those in the second-class hospitals,and the fields needing to be improved included the inter-department cooperation,personnel allocation,non-penalty reactions on mistakes.Conclusion The cognition of nursing staffs in 7 hospitals of Nanchang City on the patients safety culture is in a higher level,but insufficiency still exists,partial fields need to be continuously improved.