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

Classification of biological agents is a basic work in biodefense capability building that can help set priorities of biodefense .In this study , we compared and analyzed the Biological Agent Category Lists and the defining criteria used by the Center for Disease Control and Prevention ( CDC) of the United States, the European Union and Russia .We also compared with biological agent category lists for other purposes .

BACKGROUND:Simvastatin enhanced the expression of bone morphogenetic protein-2(BMP-2),which plays an anabolic role in bone metabolism and osteoblastic lineage differentiation.However,little is known about the molecular mechanism of simvastatin on regulation of bone marrow stromal cells differentiation.OBJECTIVE:To investigated the effect of simvastatin on osteoblastic differentiation of bone marrow stromal cells based on genetics level.METHODS:Bone marrow stromal cells derived from femur and tibia were cultured in different mediums with simvastatin or Vehicle for 7 days Following extraction and purification,mRNA was reverse-transcripted into cDNA.Fluorescence labelina was employed and the samples were then hybridized with oligonucleotide chip to screen the different genes,which were utillzed to analyze osteogenesis-related factors.Alkaline phosphatase and Von Kossa staining were performed at days 14 and 21,respectively.RESULTS AND CONCLUSIONS:At day 14,alkaline phosphatase-positive cells were more in the experimental group than control group.Von Kossa staining demonstrated that simvastatin could promote BMSCs osteoblastic differentiation and mineralization.Comparative analysis showed that 103 genes out of 22 575 rat genes had differential expression (≥2 fold or≤ 0.5 fold),and some genes were related to cell proliferation and ostoeblastic differentiation,including C/EBP δ,Cited,Ascl2,Ptpnl6,Wisp2,Tieg,etc.Simvastatin could induce osteoblastic differentiation of bone marrow stromal cells,involving in many osteogenetic-related genes.

Biosecurity is defined as effectively responding to biological damage associated with various destructive fac -tors and threats ,maintaining and protecting national interests ,security and public health in the era of globalization .This pa-per analyzes the overall situation of biosecurity capability building since the SARS outbreak in 2003 before concluding that remarkable progress has been made in China , but the gap remains great compared with the need of national security envi -ronment and the capacity building of developed countries .This paper outlines four considcrations regarding the development of biosecurity research strategic planning and offer eight tips on strengthening research on biosecurity technology in China .

BACKGROUND:Sepsis has become the greatest threat to in-patients, with a mortality of over 25％.The dysfunction of gut barrier, especially the immunological barrier, plays an important role in the development of sepsis. This dysfunction occurs after surgery, but the magnitude of change does not differentiate patients with sepsis from those without sepsis. Increased intestinal permeability before surgery is of no value in predicating sepsis. The present study aimed to observe the changes of intestinal mucosal immunologic barrier in rat models of sepsis induced by cecal ligation and puncture. METHODS:Sixty Sprague-Dawley rats were randomly divided into a sepsis group (n=45) and a control group (n=15). The rats in the sepsis group were subjected to cecal ligation and puncture (CLP), whereas the rats in the control group underwent a sham operation. The ileac mucosa and segments were harvested 3, 6 and 12 hours after CLP, and blood samples were collected. Pathological changes, protein levels of defensin-5 (RD-5) and trefoil factor-3 (TFF3) mRNA, and lymphocytes apoptosis in the intestinal mucosa were determined. In an additional experiment, the gut-origin bacterial DNA in blood was detected. RESULTS:The intestinal mucosa showed marked injury with loss of ileal villi, desquamation of epithelium, detachment of lamina propria, hemorrhage and ulceration in the sepsis group. The expression of TFF3 mRNA and level of RD-5 protein were decreased and the apoptosis of mucosal lymphocyte increased (P<0.05) in the sepsis group compared with the control group. Significant differences were observed in RD-5 and TFF3 mRNA 3 hours after CLP and they were progressively increased 6 and 12 hours after CLP in the sepsis group compared with the control group (P<0.05, RD-5 F=11.76, TFF3 F=16.86 and apoptosis F=122.52). In addition, the gut-origin bacterial DNA detected in plasma was positive in the sepsis group. CONCLUSION:The immunological function of the intestinal mucosa was impaired in septic rats and further deteriorated in the course of sepsis.

Whiplash injury is a relatively common injury of clinical acupuncture and moxibustion in the United States. The mechanism and clinical manifestation of whiplash injuries as well as its pathogenesis described in TCM were analyzed in this present article. The authors introduced the TCM syndrome differentiation of whiplash injuries and claimed that both the location and the stage of disease should be considered. For the different injury locations, the meridian musculature differentiation was applied to classify the whiplash injuries as Taiyang, Yangming, Shaoyang and Shaoyin Meridian syndromes. Considering the duration of the injury, qi stagnation and blood stasis types were classified in the acute stage and phlegm accumulation, insufficiency of the liver and kidney and qi and blood deficiencies types were classified during the chronic stage. An acupuncture protocol for whiplash injuries and typical cases were also introduced.
Acupuncture Points ; Acupuncture Therapy ; Adult ; Diagnosis, Differential ; Female ; Humans ; Middle Aged ; Whiplash Injuries ; diagnosis ; therapy

OBJECTIVETo observe the effect of dexamethasone on the proliferation and apoptosis of embryonic palatal mesenchymal (EPM) cells, and chose a proper concentration of dexamethasone which can effect the ordinary growth of embryonic palatal mesenchymal cells.

METHODSThe primary EPM cells were isolated and cultured in vitro, then we did biological assay. EPM cells were treated with different concentration dexamethasone (1 x 10(-9), 1 x 10(-8), 1 x 10(-7) and 1 x 10(-6) mol x L(-1)) respectively. The proliferation of EPM cells was evaluated using MTT method. Apoptosis was examined quantitatively with fluorescein stain.

RESULTSIn the condition of blood serum's concentration at 10%, optical density step down following the raise of dexamethasone's concentration. The effect of dexamethasone got to a summit at 3 days. Inhibition rate of dexamethasone at 1 x 10(-6) mol x L(-1) was the highest.

CONCLUSIONDexamethasone at 1 x 10(-6) mol x L(-1) can not only inhibit the growth of the EPM cells, but also will not lead to a large number of cells death. Therefore, this concentration can be used as a reference standard in future research. The most significant drug action time of dexamethason appears at the third day after administration, then the effect became weaken following the drug metabolism.

Animals ; Apoptosis ; Cell Proliferation ; Cells, Cultured ; Dexamethasone ; Humans ; In Vitro Techniques ; Mesenchymal Stromal Cells ; Mice

Palpation is one of the major diagnostic methods in Chinese medicine. The article has summed up the current status, features and origins of various palpations in different acupuncture schools in the United States, including palpation on meridian, collaterals and acupoints in traditional Chinese acupuncture, abdominal palpation of Japanese acupuncture, pulse checking for couples in five-element acupuncture, palpation on tendon straps and muscle response testing in acupuncture physical medicine. Since great importance has always been attached on social psychological factors in all acupuncture schools of America, it is thus concluded that the interactive mechanism of diagnosis and treatment represented by palpation feedback examination is not only the manifestation of people oriented individualized treatment both physically and psychologically in America, but also a kind of inheritance and development of diagnosis and treatment in the modernization of TCM.
Acupuncture ; education ; Humans ; Medicine, Chinese Traditional ; methods ; Palpation ; methods ; United States

OBJECTIVETo investigate the relation between different extents of cleft malformation with the speech characteristics in patients with cleft palate.

METHODSThe formant frequency of vowel [i] of 46 incomplete cleft palate patients (ICCP group) and 56 complete cleft palate patients (CCP group) before and after cleft palate repair, as well as 30 normal people (C group), were measured and analyzed on spectrogram.

RESULTSThe comparison of F1 between C group and CCP, ICCP before surgery showed no difference. So did the comparison of F1 between C group and CCP, ICCP after surgery. The comparison of F2 between C group and CCP, ICCP before surgery showed significant difference. The value of the C group was the highest. The value of the ICCP was higher than that of CCP. So did the comparison of F2 between C group and CCP, ICCP after surgery. The comparison of F3 between C group and CCP, ICCP(including before and after surgery) was similar to the results of F2 between the three groups. The comparison of F1 between before and after surgery in ICCP group showed no difference. However, the same kind of comparison of F2 and F3 showed significant differences: Both the values after surgery were higher than those before surgery. The comparison of Fl, F2 and F3 between before and after surgery in CCP group was similar to that in ICCP group.

CONCLUSIONThe extent of the cleft malformation is closely related to the status of the speech in patients with cleft palate. With the malformation more severe, the tongue will move backward more obviously, the elevation of the soft palate after cleft palate repair will be less active. Two ways are recommended for those patients with CCP: (1) Early interceptive orthodontic treatment to reduce the extent of palate malformation; (2) The hard palate repair can be performed prior to the soft palate repair. Patients with severe cleft lip and palate can have hard palate repaired while accepting the early cleft lip repair.

Cleft Lip ; Cleft Palate ; Female ; Humans ; Male ; Palate, Hard ; Speech

OBJECTIVETo observe the alteration of fibroblast growth factor 10 (Fgf10) and fibroblast growth factor receptor 2 (Fgfr2b) signal in mouse embryonic palate after dexamethasone and vitamin B12 exposure.

METHODSDams were divided teratogenetic group, antagomistic group and control group and were respectively injected dexamethasone, dexamethasone and vitamin B12, and normal sodium. Dams were killed and fetus was collected at embryo 12.5 and 13.5 day. The expression of Fgf10 and Fgfr2b and mesenchymal cells proliferation of mouse embryonic by western blotting and BrdU assay were checked.

RESULTSFgf10 and Fgfr2b expression was down-regulated and mesenchymal cells proliferation was inhibited significantly after dexamethasone exposure. After vitamin B12 treatment, Fgf10 and Fgfr2b expression did not restore, but cells proliferation was recovered.

CONCLUSIONDexamethasone and vitamin B12 affected the expression of Fgf10 and Fgfr2b of mouse embryonic palate and mesenchyme cells proliferation, but the change was disaccord.

Animals ; Cell Proliferation ; Dexamethasone ; Fibroblast Growth Factor 10 ; Mice ; Receptor, Fibroblast Growth Factor, Type 2 ; Signal Transduction ; Vitamin B 12