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

0.05).Seroquel showed high incidence rate of drowsiness,dizziness and weight gain(26.1%,23.9%,17.3%),chlorpromazine showed high incidence rate of extrapyramidal side effect,pyknocardia and erect collapse(60.9%,39.1%,32.6%).CONCLUSIONS:There was no significant difference of curative effect for the treat-ment of schizophrenia between seroquel-treated group and chlorpromazine-treated group,but seroque showed low side effect and high safety.

To investigate the epidemic and evolutionary trends of enterovirus (EV) in the external environment of Yunnan Province, China, molecular typing was performed on 4 EV strains that were isolated from environmental sewage in Yunnan. The VP1 region of isolates was amplified by RT-PCR using universal enterovirus primers, and the amplified VP1 region was sequenced for GenBank BLAST search and genotype analysis. The 4 EV strains were identified as ECHO7. Their nucleotide and amino acid homologies with the VP1 sequences of 68 ECHO7 strains retrieved from GenBank were measured by Mega software analysis. Our findings showed that ECHO7 strains from environmental sewage and population samples were in different evolutionary branches. These strains showed typical geographical and temporal differences; In addition, there were different transmission chains at the same time and in the same area. ECHO7 strains isolated from sewage water and patients with acute flaccid paralysis during the same period in Yunnan belonged to different clusters and evolved at different speeds. Special concerns are needed for this problem. Continuous molecular biological surveillance of human EV in the external environment of Yunnan will provide strong support for early warning of EV diseases.
China ; Databases, Genetic ; Enterovirus ; genetics ; isolation & purification ; Evolution, Molecular ; Humans ; Molecular Epidemiology ; Sequence Analysis ; Sewage ; virology

BACKGROUND:Half-ring sulcated external fixator is a bone external fixation device.Its structure is relatively simple,with multi-plane fixed,and small occupying space.So far,there are few experimental reports about construction of tibial defect model by it in large animals.OBJECTIVE:To validate the practicality and repeatability of half-ring sulcated external fixator in establishing tibial defect models in goats.DESIGN,TIME AND SETTING:The randomized controlled observation was performed at the Animal Laboratory of the Third Military Medical University from March 2005 to February 2007.MATERIALS:Nine healthy adult(Animal Center of Third Military Medical University);Half-ring sulcated external fixator,made by Professor Li of the Third Military Medical University;Osseous pin,ψ2.5 mm,Shanghai Medicai Instruments Co.,Ltd.METHODS:After animals were anesthetized,two needles were threaded in the same plane of cancellous bone about 1.5-2.0 cm above the superior articular surface of the tibia,named group 1.The angle between the two needles was 40°.One needle was threaded at the place of 3.0 to 5.0 cm under the group 1 and parallel with optional needle of the group 1,named group 2.Needles of group 4 were threaded at the cancellous bone that was 2.0 cm from the joint surface of inferior extremity of the tibia.One needle was threaded at the place of 3.0 to 5.0 cm from group 4 and parallel with optional needle of the group 4.The included angle between these two needles of group 2 and group 3 was 60°.There were totally 6 needles.Tibia was fixed by half-ring sulcated external fixator.According to the length of the tibia from X-ray,tibia and periosteum were amputated by wire saw between the two needles in the middle and inferior segment of the tibia to make segmental bone and periosteal defects of 20 percent of tibia.MAIN OUTCOME MEASURES:①Postoperative general state of the animals and effect of half-ring sulcated external fixator.②Self-repair of bone defects by X-ray and Lane-Sandhu scoring standards.RESULTS:Of the 9 selected goats,1 died of postoperative infection,and 1 developed pinhole infection and supplemented.All the others survived and were included in final analysis.The goats were awake within 2-6 hours after surgery and able to stand up to eat.The loosening nuts were screwed up.No external fixation failure or loosening was found.No skin was necrotic.Injured limb could touch the ground in 2 days,but could not bear load until 2 weeks.The goats could walk in three weeks with a slight limp,then moved freely without claudication in four weeks after the operation.At the 5th week after operation,the radiographs showed little periosteal reaction.At the 10th week after operation,sclerosis was found in the fracture ends,and medullary cavity started to close.Fifteen weeks after operation,the medullary cavity closed and periosteal reaction did not aggravate.Inaddition.Lane score was 0 at any time point.No bone formation was found in the histological examination at 5,10 and 15 weeks after operation,and Lane score was 0 at any time point.CONCLUSION:The goat model established in goat tibia with 20%defects by the half-ring sulcated external fixator shows no bone healing after 15 weeks by X-ray or histological exanimation,indicating this is a practical and repetitive method to establish animal models of large segmental defect for bone tissue engineering.

Objective To explore the effect of home-based cardiac rehabilitation training on ejection fraction, blood lipid and exercise endurance in patients with coronary heart disease. Methods Randomized controlled trials (RCTs) on the effect of home-based cardiac rehabilitation training on ejection fraction, blood lipid and exercise endurance in patients with coronary heart disease were searched via computer in PubMed, Web of Science, Embase, Cochrane Library database, Wanfang database, CNKI database, VIP database and CBMdisc. After literature screening, data extraction and quality evaluation, Revman5.3 software was used for meta-analysis. Results A total of 11 articles were included. Meta analysis showed that, compared with the conventional treatment group, home-based cardiac rehabilitation training could effectively improve patients' ejection fraction[MD=1.81,95%CI(0.68,2.95),P=0.002],exercise endurance[MD=21.97,95% CI(1.80, 42.14), P=0.03], and reduce cholesterol level[MD=-0.15,95%CI(-0.23,-0.07), P＜ 0.01]. However, there was no significant improvement on high-density lipoprotein, low-density lipoprotein, triglyceride (P＞0.05). Sensitivity analysis showed that the overall results of the study were relatively stable, and individual study had no significant influence on the overall results. Conclusions Home-based cardiac rehabilitation training can significantly improve patients' ejection fraction and exercise endurance and reduce cholesterol level, but in terms of improving patients' high density lipoprotein, low density lipoprotein and triglyceride, more large sample, multi-center long-term studies are needed to verify.

OBJECTIVETo determine the clinical characteristics and outcomes of redo CABG.

METHODSThe outcomes of 42 consecutive patients who underwent redo CABG from January 2002 to December 2010 was analyzed. There were 29 males and 13 females, aging from 46 to 78 years old with a mean of (61.2 ± 2.1) years. Off-pump CABG was applied for 31 patients and on-pump CABG for 11 patients. There were 1 patient underwent concomitant aortic valve replacement and 1 patient underwent aortic root and right aortic arch replacement respectively.

RESULTSThree patients died of right ventricle rupture, heart failure and multiple system organ failure respectively and the perioperative mortality rate was 4.8%. The post-operatively mechanical ventilation time varied from 9 to 27 h with a mean of (17 ± 7) h. There was no residual angina and perioperative myocardial infarction in the remaining patients who were all discharged uneventfully. Intraoperative 6 patients had accepted intraaortic balloon counterpulsation. During the follow-up from 6 months to 4.5 years for 38 patients, which showed no evidence of recurrent angina and postoperative coronary CT angiography in 12 patients showed the patency of grafts is good.

CONCLUSIONSatisfactory outcome of redo coronary artery bypass grafting can be achieved if proper indication were choose and reasonable management were performed.

Aged ; Cardiopulmonary Bypass ; Coronary Artery Bypass ; Coronary Restenosis ; surgery ; Female ; Humans ; Male ; Middle Aged ; Reoperation ; Treatment Outcome

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 investigate the expression and clinical significance of mismatch repair genes hMLH1 and hMSH2 in sporadic colorectal carcinoma tissues.

METHODSThe expression of hMLH1 and hMSH2 proteins was detected in the 63 sporadic colorectal carcinoma samples by immunohistochemical staining, including tumor tissue, adjacent tissue at 3 cm from the carcinoma, and normal tissue at 10 cm away from the tumor.

RESULTSThe positive rate of hMLH1 protein expression in the 63 normal colorectal tissues, adjacent tissues and sporadic colorectal carcinoma tissues was 95.2%, 85.7% and 81.0%, respectively. The positive rate of hMLH1 protein expression was significantly lower in the tumor than in normal colorectal tissues (P < 0.05). The positive rate of hMSH2 protein in the 63 normal colorectal tissues, adjacent tissues and sporadic colorectal carcinoma tissues were 76.2%, 66.7% and 52.4%, respectively. The positive rate of hMSH2 protein expression was significantly lower in the tumor than in normal colorectal tissues (P < 0.01). The positive rate of hMLH1 protein expression was significantly higher in the tumor tissue of patients aged younger than 60 years (100%) than that in patients ≥ 60 years (75.0%, P < 0.05). The positive rate of hMLH1 protein expression in the tumor tissue accompanied by lymphatic metastasis was 50.0%, significantly lower than that (93.3%) in tumors without lymphatic metastasis (P < 0.05). The positive rate of hMSH2 protein expression in the tumor tissue of patients aged younger than 60 years was 80.0%, significantly higher than that (43.8%) in the cases ≥ 60 years (P < 0.05). The positive rate of hMSH2 protein expression in the tumor tissues with invasion reaching to the intestinal serosa (61.5%) was significantly higher than that (37.5%) in the tumors invading to submucosa or muscular layer (P < 0.05). There was a positive correlation between the expressions of hMLH1 and hMSH2 proteins in the sporadic colorectal carcinomas.

CONCLUSIONThere is a certain loss of expression of hMLH1 and hMSH2 proteins in sporadic colorectal carcinoma, and is correlated with the age of patients, lymphatic metastasis and different depth of cancer invasion. HMLH1 and hMSH2 may be used as a useful laboratory marker in clinical judgement of occurrence and development of sporadic colorectal carcinoma.

Adaptor Proteins, Signal Transducing ; metabolism ; Adenocarcinoma ; metabolism ; pathology ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; metabolism ; Colonic Neoplasms ; metabolism ; pathology ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; metabolism ; Neoplasm Invasiveness ; Nuclear Proteins ; metabolism ; Rectal Neoplasms ; metabolism ; pathology

OBJECTIVEThis report presented an overview on the epidemiology of enterovirus in Yunnan province, the People's Republic of China.

METHODSA total of 210 strains of non-polioviruses isolated under acute flaccid paralysis surveillance during a 5-year study period from 1997 to 2000 and 2004 were examined. Of the 210 non-polioviruses strains, a total of 12 strains of adenoviruses were serologically identified. The remaining 198 isolates were used for molecular typing, and the viral genomes of 195 nonpolio enteroviruses (NPEVs) were translated to corresponding amino acid sequences and compared with those of the prototype strains.

RESULTSBased on molecular typing, 5 isolates were classified into 5 serotypes of human enterovirus A species while 158 isolates into 34 serotypes of B and 32 isolates into 6 serotypes of C species. However, we did not isolate any viruses which belonged to human enterovirus D species. Thus, under acute flaccid paralysis surveillance, human enterovirus B species accounted for 75.2% of the 210 isolates and was considered as the predominant one, followed by human enterovirus C (12.2%), adenovirus (5.7%), and human enterovirus A (2.4%).

CONCLUSIONAlthough the epidemiological characteristics of NPEVs from Yunnan province remained "unknown", the molecular typing method had provided us a breakthrough to understand the epidemiology of these viruses.

China ; epidemiology ; Enterovirus ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; epidemiology ; Genes, Viral ; Humans ; Molecular Epidemiology ; Serotyping

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