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 compare the mental health between the urban mothers and rural mothers of children with cleft lip and pal-ate.Methods:1 68 mothers(including 1 24 rural and 44 urban)of the children with cleft lip and palate were included in the trial group and 82 mothers of normal children were included in the control group.The subjects were tested by the self-assessment symptom lists of SCL-90 for the evaluation and comparison of mental health.Results:The scores of nine dimensions of trial group in somatization, forcing symptoms,depression,anxiety,etc were significantly higher than those of the control group(P <0.05).The scores of nine di-mensions of rural group in somatization,forcing symptoms,depression,anxiety,etc were higher than those of the urban group,the ri-val,psychotic and total scores showed significant differences(P <0.05);Conclusion:The mental health status of mothers of children with cleft lip and palate is poorer than that of those of normal children.That of the rural mothers is poorer than that of the urban ones. Special attention should be paid to the mental health status of the parents of children with cleft lip and palate.

Objective:To investigate the clinical effect of Niaodukang-enema liquid on chronic renal failure. Methods: 112 cases were divided into treatment group and control group randomly. The patients of treatment group were given oral Medicinal Charcoal Tablets and Niaodukang-enema liquid(120ml) for enema at high position,56 cases. The patients of contral group were given oral Medicinal Charcoal Tablets only, 56cases. The renal function was observed before and after treatment in both groups.Results :The distinct effective rates and total effective rates in treatment group were obviously better than that in contral group (P

It was widely believed that the interaction between rice and rice blast fungus can be interpreted by the gene-for-gene hypothesis. Two interaction models between rice blast fungus and anti-disease genes had been briefed. They were receptor-ligand model and guard model. The progress of research about molecular marker and position on avirulence genes of the pathogens of rice blast (Magnaporthe oryzae) was reviewed, and the methods to clone and the cloned avirulence genes of rice blast fungus were also summarized.

OBJECTIVETo observe application value of multispectral animal living imaging technology in rats model of osteoarthritis.

METHODSFifteen male SD rats weighed (180 +/- 20) g (3 months old) were received intra-articular injection of iodoacetic acid for establishing osteoarthritis. Articular cavity of left knee of rats were injected into 50 microl iodoacetic acid. The same volume of sterile saline was injected into right knee articular cavity as control. X-ray living imaging and bone mineral density were observed at 2 and 4 weeks after establishment of model. After 4 weeks,rats were sacrificed and their bilateral joints were collected and determined histologically based on Collins classification and Kellgren-Lawrence classification.

RESULTSOsteoarthritis model was successfully established, compared with control group, model group showed typical manifestation of osteoarthritis, including irregular cartilage surface,osteophyte formation,joint deformity and cartilage defect,and combined with significant decrease of bone density (P < 0.01), while the decrease was not obvious in proximal tibia (P < 0.05). After 2 weeks, knee joints in model group was classified as Collins grade 1 and Kellgren-Lawrence grade 2,then classified as Collins grade 4 and Kellgren-Lawrence grade 3 after 4 weeks,control group showed smooth articular surface,normal joint space and intact cartilage surface, knee joints was classified as Collins and Kellgren-Lawrence grade 0, and bone density of distal femur and proximal tibia were normal.

CONCLUSIONMultispectral animal living imaging technology could be used in dynamic observation of living imaging and detection of bone density in the animal model of osteoarthritis, and it is significant for evaluation of osteoarthritis model, and its realted tesearch.

Animals ; Bone Density ; Disease Models, Animal ; Humans ; Knee Joint ; diagnostic imaging ; Male ; Osteoarthritis ; diagnosis ; diagnostic imaging ; physiopathology ; Radiography ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the effect of dexmedetomidine as a supplementary to combined lumbar plexus-sciatic nerve block for arthroscopic knee surgery.Methods Fifty patients who were scheduled for arthroscopic knee surgery and ASA class Ⅰ or Ⅱ grade,were divided into control group and dexmedetomidine group by random digits table method with 25 cases each.The patients of dexmedetomidine group were given loading dose dexmedetomidine 0.8 μ g/kg(injection time 10 min) by intravenous infusion,then were given maintenance dose 0.4 μμ g/(kg· h) to the end of operation.The patients of control group were given 0.9％ sodium chloride,methods with the dexmedetomidine group.After 10 min,combined lumbar plexus-sciatic nerve block was performed by a nerve stimulator in both groups.Fentanyl 50 μμ g was infused intravenously to the patient whose effect of block was inadequately.The mean arterial pressure (MAP),heart rate (HR),pulse oxygen saturation (SpO2),respiratory frequency at preanesthesia (T0),beginning of dexmedetomidine infusion (T1),beginning of surgery (T2),10 minutes of surgery (T3) and the end of surgery (T4) were recorded,and intraoperative untoward reaction,using of fentanyl and doctor's satisfaction rate were recorded.Results There were no statistical differences in MAP and HR at T0 between the 2 groups (P＞ 0.05).MAP and HR at T2-4 were significantly higher than those at T0 in control group,furthermore MAP and HR at T2-4 in control group were significantly higher than those in dexmedetomidine group [MAP:(137 ± 18) mm Hg(1 mm Hg =0.133 kPa) vs.(107 ± 14) mm Hg,(132 ± 11) mm Hgvs.(107 ± 11) mm Hg,(131±13) mm Hg vs.(95 ± 12) mm Hg;HR:(99 ± 17) times/rmin vs.(88 ± 14) times/min,(99 ± 14) times/min vs.(81 ± 15) times/min,(97 ± 14) times/rmin vs.(76 ± 11) times/min],there were statistical differences (P ＜ 0.05).There were no statistical differences in respiratory frequency and SpO2 between the 2 groups (P ＞0.05),but 2 cases in dexmedetomidine group,5 cases in control group appeared SpO2 ＜0.90,they quickly returned to normal (＞ 0.97) by holding up the submaxilla.The using rate of fentanyl and doctor's satisfaction rate in dexmedetomidine group were significantly better than those in control group [28％(7/25)vs.68％(17/25),100％(25/25) vs.80％(20/25)],there were statistical differences (P＜ 0.05).There were no statistical differences in the using rate of atropine and efedrina,the rate of respiratory depression between the 2 groups (P ＞ 0.05).Conclusion Dexmedetomidine as a supplementary to combined lumbar plexus-sciatic nerve block for arthroscopic knee surgery has effect confirmed,low untoward reaction,high doctor' s satisfaction rate.

Objective To investigate the effect of folic acid combined with Helicobacter pylori (Hp) eradication therapy on chronic atrophic gastritis.Methods From December 2009 to March 2011 at the First Affiliated Hospital of Nanjing Medical University,184 patients with endoscopic and pathological diagnosis of chronic atrophic gastritis (90 Hp positive and 94 Hp negative) were selected.Hp positive patients were divided into group A and group B.Forty-three patients in group A were treated with standard triple Hp eradication therapy and follow by folic acid therapy for three months.Forty-seven patients in group B and Hp negative patients received three months of folic acid therapy.The clinical symptoms of each group were scored before treatment,one month after folie acid therapy and three months after folic acid therapy and analyzed by t test. Patients of each group received gastroscopy before treatment and three months after medicine withdrawal. Endoscopic scores,pathological scores and t test were recorded.The serum levels of pepsinogen ( Ⅰ,Ⅱ ) and gastrin 17 in venous blood of 55 Hp negative patients were detected by enzyme-linked immunosorbent assay (ELISA) method before treatment and three months after medicine withdrawal.Results Compared with three months therapy (1.15 ± 0.03),after one month folic acid therapy (1.55 ± 0.04) was statistically significant in clinical symptoms score of all patients (t =8.18,P＜0.01).By the end of therapy,clinical symptom score of group A (1.06 ± 0.04) was lower than that of group B (1.56 ±0.08),and the difference was significant (t=6.00,P＜0.01).There was significant difference in endoscopic scores of all patients between before treatment (1.57±0.95) and after treatment (1.00±0.76,t=11.12,P＜0.01).The differences in each pathological score of all patients (inflammatory scoring,active scoring,atrophy scoring,intestinal metaplasia scoring,atypical hyperplasia degree scoring) were significant between before treatment and after treatment (t=5.51,6.90,7.53,6.34,2.90,respectively,all P＜0.01).The serum level of pepsinogen Ⅰ before treatment of 55 Hp negative patients [(1.03±0.19) nmol/L] was lower than that after treatment [(2.24±0.33) nmol/L],and the difference was statistically significant (t=3.19,P＜0.01).After treatment the level of gastrin 17 [(0.86±0.05) nmol/L] was higher than that before treatment [(0.47±0.05) nmol/L],and the difference was statistically significant ( t =5.33,P＜ 0.01 ).Conclusion Folic acid in combination with Hp eradication therapy can be favorable to atrophic gastritis,which may promote the secretion of pepsinogen and gastrin.

OBJECTIVETo observe the intervention effects of electroacupuncture at "Yishu" (EX-B 3) on rats with type-2 diabetes mellitus (T2DM), so as to provide experiment references for acupuncture to treat T2DM.

METHODSAmong seventy male Wistar clean-grade rats, 8 rats were randomly selected into a control group; the rest rats were made T2DM model. Fifty-two rats which were successfully made T2DM model, according to randomized block method, were divided into a model group (10 rats), a medication group (10 rats), an electroacupuncture at "Shenshu" (BL 23) group (11 rats), an electroacupuncture at "Pishu" (BL 20) group (10 rats) and an electroacupuncture at "Yishu" (EX-B 3) group (11 rats). Seven days after successful establishment of model, the rats in the model group were fixed in the self-made rat bag without receiving any treatment; the rats in the medication group, according to body mass (10 mL/kg), were treated with intragastric administration of glimepiride; the rats in all the electroacupuncture groups were treated with electroacupuncture at "Shenshu" (BL 23), "Pishu" (BL 20) and "Yishu" (EX-B 3), respectively. The continuous wave was selected with a frequency of 15 Hz and a current intensity of 4 to 6 mA. The treatment was given 20 min per treatment, once a day, 5 treatments per week for continuous 4 weeks. Before the establishment of model and continuous 4 weeks after the intervention, blood samples were collected from rats' caudal vein, and fasting blood glucose (FBG) was measured with FBG device each week. After the last intervention, the rats were killed and hypothalamus, pituitary and adrenal gland were collected. The colorimetric method was applied to measure the contents of triglyceride (TG), cholesterol (TC), high-density lipoprotein (HDL-C) and low-density lipoprotein (LDL-C); radioimmunoassay was used to test the contents of glycated serum protein (GSP), fasting insulin (FINS), corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortin (CORT).

RESULTSFour weeks after the intervention, except that the rat's body mass in the normal group continued to increase, body mass in the model group, medication group and each electroacupuncture group were significantly reduced (P<0.01, P<0.05). Compared with the model group, the FBG in the electroacupuncture at "Pishu" (BL 20) group and electroacupuncture at "Yishu" (EX-B 3) group were obviously reduced (P<0.05, P<0.01); FBG in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (both P<0.05). The contents of TG, HDL-C and LDL-C in the electroacupuncture at "Yishu" (EX-B 3) group were reduced (P<0.05, P<0.01), the content of TG was significantly lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (both P<0.05), the content of LDL-C was significantly lower than that in electroacupuncture at "Shenshu" (BL 23) group (P<0.05). Insulin sensitivity index (ISI) in the medication group, electroacupuncture at "Pishu" (BL 20) group and electroacupuncture at "Yishu (EX-B 3)" group were evidently increased (P<0.05, P<0.01); ISI in the medication group was lower than that in the electroacupuncture at "Yishu" (EX-B 3) group (P<0.05). The content of CRH in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (P<0.05, P<0.01); the content of CORT in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Pishu" (BL 20) group (both P<0.05).

CONCLUSIONElectroacupuncture at "Yishu" (EX-B 3) could reduce the level of CORT to improve the insulin resistance in rats with T2DM, improve insulin sensitivity index, regulate blood lipid metabolism and relieve the hyperactivity of the HPA axis.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; metabolism ; therapy ; Electroacupuncture ; Hormones ; metabolism ; Humans ; Hypothalamus ; metabolism ; Male ; Pituitary-Adrenal System ; metabolism ; Rats ; Rats, Wistar

OBJECTIVE To discuss the clinical traits,pathogenesis and TCM stepwise treatments of fungal urinary tract infection.METHODS According to the risk factors and clinical character of fungal urinary tract infection,we clarified the mechanism of the disease.The principal aspect was spleen-kidney vacuity detriment and the secondary incidental was accumulated damp-heat and static blood in the lower burner,viz weaken healthy qi and excessive pathogenic factor.Hence during the clinical treatment we should regulate faculty condition.In the acute infection period we should give priority to dispel evils supplemented by the recovery of right qi.In the convalescence we should pay more attention to support right supplemented by dispelling.RESULTS The most common pathogen of fungal urinary tract infection was Candida albicans.The TCM stepwise treatments of fungal urinary tract infection together with regulatiy entire faculty condition had the characteristics of high efficacy and few side effects.CONCLUSIONS The TCM stepwise treatments of fungal urinary tract infection has more potentiality which deserves further study.

Objective To discuss the value of pregnantal ultrasonography(US) on fetus' complicate congenital heart diseases(CHD). Methods The uhrasonographic fetures of fetus complicate CHD were analyzed and compared with the pathological diagnosis. Resells Twenty fetus' complicate CHD were diagnosed by pregnantal US. Four cases with single ventricle, two eases with transposition of great arteries,two eases with double outlet right ventricle,two cases with truncus arteriosus,three cases with hypoplastic left heart syndrome, three cases with endocardial cushion defect, two cases with hypoplastic right heart syndrome,one case with tetralogy of Fallot and one case with pulmonary atresia were included. The diagnosing according rate was 100%. Conclusions Pregnantal US can fully evaluate the fetus' complicate CHD.