Objective:To observe the effect of dissection and reconstruction on palatal muscles morphology in cats. Methods: Nine cats were randomly divided into three groups according to the extension of muscle dissection: small area group(<1/3 group),medium area group(1/3-2/3 group)and large-scale area group(>2/3 group).Control group was the normal palatal muscle in any normal side of the palate.Palatal muscles of all experimental groups were dissected from the posterior border of the hard palate and were reconstructed later. The morphology features and ultrastructure of palatal muscles were observed respectively by HE staining and transmission electron microcopy 1-3 month after operation. Results: There were no obvious differences in morphologic features and ultrastructure of palatal muscles between experiment and normal control groups when the range of palatal muscle dissection was less than 2/3. At early stage (1 month) after operation in large-scale area group, the arrangement of muscles fiber were in structural disorder with inflammatory cell infiltration and muscle fibril formation. The Z line was not distinct, muscles fiber became narrow and myocomma arrangement was not fully dense under transmission electron microcopy. At later stage (2 or 3 month after operation) in large-scale area group, the arrangement of muscles fiber were comparatively regular, neonatal muscles fibers became gradually mature and the amount of inflammatory cell had obviously decreased. Ultrastructure study showed that myocomma arrangement was more orderliness, and Z line became more distinct. Conclusion: It suggests that the extensive dissection of palatal muscles carried no fibrosis, and injury musculature could be repaired and regenerated.

Objective　To study the histologic source, clinical features and treatment methods of leiomyoma of the vagina. Methods　From January of 1988 to January of 1999, 25 patients with leiomyoma of the vagina were retrospectively analyzed. Results　The clinical features of leiomyoma of the vagina were slow in growth and solitary in number. Leiomyoma of the vagina can be recurrence and sarcomatous change. The symptoms of leiomyoma of the vagina depended on the size and location of the leiomyoma. Treatment consisted of surgical excision by vagina. Conclusions　Leiomyoma of the vagina is a rare condition. Whenever such a tumor is detected, it has to be removed immediately to prevent further growing and sarcomatous change in the future.

Objective To investigate the possible existence of HBoV in children with acute respiratory infections in Nanjing area and explore its relationship with clinical characteristics.Methods A total of 397 nasopharyngeal secretion samples were collected from children with acute respiratory infection,admitted from July 2009 to June 2010 in Nanjing Children'S Hospital affiliated to Nanjing Medical University,and 50 cases of children without symptoms of respiratory infection were recruited as control group,whose nasopharyngeal secretion samples were also collected.HBoV was determined by real-time fluorescence quantitative PCR.MP and CT were detected by real-time fluorescence quantitative PCR in those HBoV-positive samples.RSV,ADV,IVA,IVB,PIV-1,PIV-2,PIV-3 and hMPV were detected by direct antigen-specific immunofluorescence assays.HBoV NP-1 fragments were amplified and sequenced in 5 HBoV positive samples randomly selected.The results were compared with the known GenBank sequence,and thereby the phylogenetic tree was established.The epidemiological characteristics,clinical presentation and the final clinical diagnosis of HBoV were analyzed according to the clinical data of the HBoV-positive patients.Results Thirty-three HBoV-positive cases were detected by real-time fluorescence quantitative PCR method with a positivity rate of 8. 3% ( 33/397 ). Among the 33 HBoV-positive cases, 19 cases (57.6%) were multiple infections with HBoV and other pathogens, the top three of which were MP (27.3% ,9/33 ),RSV (24.2% , 8/33 ) and PIV-3 ( 12. 1% ,4/33 ). Affected children aged from 7 to 36 months old accounted for 75.8% of the total ( 25/33 ). The measured HBoV NP-1 gene sequences of 5 specimens were consistent,indicating a high homology (99% to 100% ) with the stl, st2 and WHL-1. Conclusions HBoV is one of the pathogens of children's acute respiratory infections in Nanjing. HBoV NP-1 gene is highly conserved,with little variation in different seasons and in different regions and therefore can be used as a marker for real-time fluorescence quantitative PCR and other methods.

Objective To observe the roh of cytoimmunologic factors,careinoembryonic antigen (CEA),serum ferritin(SF)and prognostic factors in patients with Iung adenous cancer.Methods Eightyone patients with lung adenous cancer were analyzed retrospectively.The clinical features as well as the alternation of T-lymphocyte subsets and its relationship with CEA.SF,disease stage and metastasis of lymph node were studied.Results(1)The percentage of CD4+ T cell decreased and that of CD8+T cell increased,the ratio of CD4+/CD8+ went down accompanied by the increase of disease stage.CEA and SF increased as the dmease stage increased.(2)Thedecrease of percentage of CD4+ T cell and the ratio of CD4+/CD8+ was obvious in patients with metastasis of lymph nodes[LN(+)](p＜0.01).The increase of percentage of CD8+Tcell and CEA,SF level was obvious in LN(+)(P＜0.01 or＜0.05).(3)The decrease of percentage of CD4+T cell and the ratio of CD4+/CD4+ was obvious in CEA (+)(P＜0.01).The increase of percentage of CD8+ T cell and SF level was obvious in CEA (+)(P＜0.01).(4)The decrease of percentage of CD4+ T cell and the ratio of CD4+/CD8+ was obvious in SF(+)(P＜0.01).The increase of percentage of CD3+ CD8+ T cell and the CEA level was obvious in SF (+)(P＜0.05 or＜0.01).Conclusion Disorder of cytoimmunologic situation exists in patients with lung adenous cancer,and it is closely correlatod with the CEA,SF level disease stage and metastasis of lymph node.

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.

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

To obtain a most appropriate method for the evaluation of the three basic skills of clinical doctors,we designed an objective structured clinical examination(OSCE) model with standardized patients and applied it to the "three-basic" examinations among 50 doctors from various clinical departments.The results showed that this model provided a new way for the effective,objective and reliable evaluation of doctors' clinical knowledge and skills.

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 understand the immune regulatory function of monocyte-derived dendritic cells (MoDC) in patients with chronic severe hepatitis B (CSHB) and its roles in the severe illness progression of chronic hepatitis B (CHB) by detecting surface phenotype of MoDC and expression level of cytokines in MoDC after polyl : C treatment. Methods The peripheral blood mononuclear cells (PBMC) were isolated by Ficoll density gradient separation from 37 patients with CSHB, 20 patients with CHB, and 20 healthy controls (NC). Purified PBMC were acquired using immunomagnetic anti-CD14-beads. Then PBMC were induced to immature dendritic cell (iDC) in vitro. PolyI : C was added to induce DC maturation. The mean fluorescence intensity (MFI) of the phenotype marker molecules including HLA-DR, CD83, CD86 and CD80 on surface of iDC and mature DC (mDC) were detected by flow cytometry. The supernatants of MoDC culture were collected at 12,24 and 48 h after polyI : C treatment, respectively and the release levels of interleukin (IL)-12, IL-6and tumor necrosis factor (TNF)-α were determined by enzyme linked immunosorbent assay (ELISA). Comparisons among groups were done by single factor analysis of variance and homogeneity of variance was tested. Results There were no significant differences of phenotype marker molecules on cell surface of iDC, including HLA-DR, CD83, CD86 and CD80 in CSHB, CHB and NC groups.However, the expressions of HLA-DR, CD83, CD86 and CD80 on cell surface of mDC in CSHB group were lower than those in CHB and NC groups (F=59.73, 13.95, 34.80 and 73.02, respectively; all P＜0. 05). The secretions of IL-12 at three time points of 12 h, 24 h and 48 h after polyI : C treatment in group NC were higher than those in CHB and CSHB groups (F= 151.34, 126.65 and 72.76, respectively; P＜0.05), and peaked at 24 h which were (48.2±7.6), (56.7±11.8) and (97.8±16.2) ng/L, respectively. The secretions of IL-6 at the above three time points were CSHB＞CHB＞NC (F=92.50, 86.89 and 64.57, respectively; all P＜0. 05) and peaked at 12 h which were (1698.3±340.4), (965.8±231.7), (697.8±213.6) ng/L, respectively. The secretions of TNF-αat the above three time points were CSHB＞CHB＞NC (F=58.66, 122.36 and 44.73, respectively;all P＜0. 05) and were (19 672. 7±4214. 7), (9946. 1 ± 2586 5), (6659. 2±955. 8) ng/L,respectively at 24 h after treatment. Conclusions MoDCs of CSHB patients show mature defection and abnormal cytokine secretion. The expression level of IL-12 which mediates cellular immune is low.Meanwhile, the productions of IL-6 and TNF-α which mediate inflammatory response are up-regulated. This may be one of the major factors which lead to exacerbation of liver inflammation and ultimately development of severe hepatitis.

Objective To observe the mobility of levator veli palatini muscle during speech in patients with repaired cleft palate and cleft lip. MethodsMRI of the levator veli palatini muscle was taken during speech and breathing in three groups of patients: control group (cleft lip only, 8 cases ),velopharyngeal incompetence (VPI)group (7 cases), velopharyngeal competence (VPC) group (10 cases ).The length and the angle of the levator veli palatini muscle were compared. ResultsThe changes of the length and the angle during speech were not significantly different among the three groups ( P ＞ 0. 05 ). The ratio of length changes when speaking "a"," I", and "m" are ( 13.5 ± 11.7) %, ( 11. 1 ±10. 8) %, (8.2 ±14. 3) %. Mean angle of pronouncing "a"," I", and "m" are [ (43. 18 ± 4. 984) °, (43.08 ± 4. 879) °,(39. 48 ±5. 046)°]. The levator veli palatini muscle contracted progressively form "m" ," I", to "a".ConclusionsThe mobility of the levator veli palatini muscle in patients with repaired cleft palate and cleft lip only is basically the same.