Background Identifying and testing of pain is very necessary for the care and decrease of the suffering of experimental animal in medical experiment.Effective method for testing the pain and distress status of experimental animal with eye disease is still absent in China.Objective This pilot study was to establish an evaluating system for assessing the pain and distress status of bullous keratopathy rabbit model.Methods This study was approved by the Ethic Committee of Beijing University First Hospital.Twelve healthy New Zealand rabbits were selected in this experiment.Bullous keratopathy model was established in the left eyes of 9 rabbits by scraping corneal endothelium as the experimental team,and other 3 rabbits were served as the control team.The cornea lesion was examined by manipulate slit lamp,and the central cornea thickness (CCT) was measured by ultrasound biomicroscopy (UBM).Weight+20 Indexes For Pain and Distress Status Referring Guidelines for Pain and Distress in Laboratory Animals made by International Animal Care and Use Committee (IACUC) were assessed and measured as well.Results Corneal edema and opacity were obvious 1 day after surgery.Corneal bullous appeared from the third day after surgery,and cornea erosion was seen at the location of bullous breakage.The corneal lesions remained until 14 days after surgery.CCT value was (1468±100),(2313±588),(2391±271) and (2362±151) μm,respectively in day 1,3,7 and 14 after the establishment of models,which showed significant increase in comparison with the preoperative CCT (390±6)μm (all P=0.000).However,no significant difference was seen in the CCT between day 3,7 and 14 (P＞0.05).Body weight of the rabbits was (3.29±0.20),(3.20-0.17),(2.77±0.25) and (3.10±0.30)kg respectively in day 1,3,7 and 14 after operation,with significant decrease in comparison with the pre-operative weight (3.52-0.18)kg in the experimental team (P=0.008,0.007,0.003,0.004).The scores for pain and distress status of all rabbits in pre-operation of the experimental team and in the control team were zero,and the score was 7 (7,7),11 (10,12),9 (8,10),9 (9,9)in day 1,3,7 and 14 in the experimental team after surgery,with the highest score in day 3,which was bullous and bullous breakage duration.Seven of twenty indexes,including the reduce of diet and drinking,self-imposed isolation/hiding,grinding teeth,aggression,deceased movement,abnormal posture,vocalization occurred in the model animals after surgery.Conclusions Weight+20 Indexes For Pain and Distress Status is an effective,impersonal and quantitative method for observing and evaluating the pain and stress status in bullous keratopathy rabbit.

The article explored the feasibility,implementation procedure,effects evaluation,existing problems and future improvements on introducing conditional-opening-experiment(COE) in experiment teaching of hygiene analysis.The results indicated that COE could culture the practical operation ability,cooperation ability,overall situation consciousness and responsibility of students in many ways,although it still needed improvements.It promoted students'study enthusiasm,initiative consciousness and their participation and strengthened the harmony relationships between students and teachers.

Objective To study the prenatal genetic diagnostic methods for hemophilia A fetus.Methods From 2002 to 2006,19 hemophilia A families were diagnosed either by long distance-polymerase chain reaction(LD-PCR)for factor Ⅷ intron 22 inversion or by the DNA polymorphism genetic linkage analysis of factorⅧin the Beijing Chaoyang Hospital.Results (1)Totally 19 women,with 22pregnancies received the prenatal diagnosis of fetal hemophilia A.The average week at diagnosis was 23(17-34)weeks.All the direct fetal blood sampling(DFBS)were successful.There was no fetal-loss caused by the procedures.(2)Of the 19 hemophilia A families,14 appeared to be factorⅧintron 22inversion,in which 16 prenatal diagnoses were done,10 fetuses were diagnosed as genetical hemophilia A patients,and 6 fetuses were normal.(3)Using combined polymorphism genetic linkage analysis 6 prenatal diagnoses were done,including one woman's two pregnancies,in which both her fetuses were diagnosed as genetical hemophilia A patients.(4)Factor Ⅷ levels of 16 fetuses were measured,and 6 fetuses were unmeasured either because the pregnancy weeks were lower than 20 weeks or the parents refused.Factor Ⅷlevel ranged from 0 to 198%.There were 11 fetuses whose factor Ⅷ levels were lower than 10%.Ten of them were diagnosed to be genetical hemophilia A patients,and in only one boy the factorⅧlevel was 2%,but the genetic diagnosis was normal and for one year's follow up he was doing normal.Conclusion LDPCR combined with polymorphism genetic linkage analysis enables a quick and correct detection ofhemophilia A carrier.For a carrier pregnancy.prenatal diagnosis could be done for the male fetus.Factor Ⅷ deficiency of the fetus could help make the diagnosis but the final diagnosis should be based on genetic evidence.

AIM: To compare the evaluations for the structure and function of the hypertrophic hearts induced by volume overload or pressure overload in rats. METHODS: Volume overload-induced cardiac hypertrophy was established by abdominal aortacaval fistula (ACF) and pressure overload-induced cardiac hypertrophy was developed by constriction of aorta (CA). The cardiac structure and function were analyzed by echocardiography, hemodynamic determination, heart weight measurement and histological examination. RESULTS: Heart weight of rats in all the operated groups was increased compared to the sham-operated groups. In 1-week ACF group, the internal diameter (0.67?0.03)cm vs (0.60?0.02)cm, P

Adult ; Female ; Food Industry ; manpower ; Humans ; Male ; Middle Aged ; Smoking ; epidemiology ; Thiocyanates ; urine

Rhubarb is a traditional Chinese medicine (TCM), wildly used in treating the disease of hyperlipidemia. However, its components are complicated, so that it is still difficult to clear the specific roles of its various components in blood lipids regulation in. So we decide to systematically study the anti- hyperlipidemia mechanism of rhubarb. We integrated multiple databases, based on entity grammar systems model, constructed molecular interaction network between the chemical constituents of rhubarb and hyperlipidemia. The network includes 231 nodes and 638 edges. Thus we infer the interactions of active targets and disease targets to clarify the anti-hyperlipidemia mechanism. And find that rhubarb can promote excretion of cholesterol; inhibit clotting factors and improve blood circulation; inhibit the release of inflammatory cytokines and maintain fat metabolism balance; inhibit cholesterol and triglyceride synthesis; and other ways to achieve lipid-lowering effect. Thus this study provides reference for novel drug development and component compatibility, and also gives a new way for the systematically study of acting mechanism of traditional Chinese medicine.
Animals ; Databases, Factual ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Gene Regulatory Networks ; drug effects ; Humans ; Hyperlipidemias ; drug therapy ; genetics ; metabolism ; Hypolipidemic Agents ; administration & dosage ; chemistry ; Lipid Metabolism ; Rheum ; chemistry ; Signal Transduction ; drug effects

Objective To study the polymorphisms of short tandem repeat (STR) loci in intron 1, 24, 13 and 22 (STR 1,24, 13, 22) of factor Ⅷ (FⅧ) gene in Chinese population, and establish single tube multiple fluorescent PCR method for rapid diagnosis of haemophilina A(HA). Methods Four STRs from genomie DNA of 220 females without blood relationship were amplified in a single tube using quadri-fluorescence PCR. Capillary electrophoresis was analyzed in ABI PRISM 310 Genetic Analyzer. DNA sequencing was used to assay the number of dinueleotide repeats. Gene diagnosis were performed in 96 HA families. Results It was observed that 7, 9, 10 and 7 different alleles were found in STR1, 24, 13 and 22, respectively. The PIC (polymorphism information contents) were 0. 3789, 0. 4055, 0. 5239 and 0. 4713 in STR1,24, 13 and 22, respectively, and the HR (heterozygesity rate) were 34. 55% (76/220), 38. 18% (84/220), 49. 55% (109/220) and 43.64% (96/220). In 96 HA families, the diagnosis rate of STR1, 24, 13 and 22 were 38. 54% (37/96), 38. 54% (37/96), 54. 17% (52/96), 42. 71% (41/96), respectively. Whereas it achieved 79. 17% (76/96) when combining the four STR markers. Conciusion The single tube multiple fluorescent PCR of four STR loci is an effective, simple, quick method for linkage analysis and gene diagnosis of haemophilia A.

Diagnosis, Differential ; Humans ; Parathyroid Glands

Objective To develop a technique for the detection of genotypes of ?-thalassemia, which was rapid and automatical with high through-put.Methods The Real Time PCR and dissociation curve analysis (D.C) were carried out by using SYBR Green1 on ABI7000.Positive results of the 3 SYBR-Q-PCRs were defined by the -dF/t℃≥cut off value of the peak at the specific Tm for each right PCR product.Three PCR products were recombined with the T-vector.The correct positive clones were selected by sequencing.Standard curves of the CT vs. log values of copies were done from a serious dilutions of the recombinant plasmid DNA which served as the template in SYBR-Q-PCRs.Results The conditions of the PCR including concentrations of primers, thurmocycler program etc.were optimized.Specific Tm values for the 3 SYBR-Q-PCRs were 82.5?1℃, 83.0?℃ and 84.0?1℃ respectively. The standard curves for p800 bp and p206 bp DNA showed a good linear relationship between CT and log value of the copies of the template DNA ranging from 10~5 copies to single copy. Sensitivity of the technique were at least 10~1 to 10~2 times higher than that of the regular PCR plus gel electrophoresis method.The techniques and reagents showed very good reproducibility and stability besides the high sensitivity. The ?-thalassemia 1(??/--SEA), Bart′s Hydrops Syndrom (--SEA/--SEA), deletion type and non-deletion type of HbH diseases, and homozygote of ?-thalassemia-2 were diagnosed by the methods.Conclusion The genotypes of the ?-Thalassemia could be diagnosed by the Real-time PCR with SYBR Green1 combined with the melting curve analysis.rapidly, automatically, accurately with high throughput and without dual fluorescent labeled probe.

OBJECTIVETo investigate the variation of serum thyroid transcription factor-1 (TTF-1) in different patients and explore its significance in the diagnosis of lung carcinoma.

METHODSDot-enzyme linked immunosorbent assay (dot-ELISA) and Leica Q500 MC image analysis system were used to quantitatively detect TTF-1 protein in the serum samples from normal healthy adults and from patients with benign lung disease, lung cancer, thyroid carcinoma and non-thyroid carcinoma.

RESULTSThe sensitivity, specificity, standardized positive predicative value, standardized negative predicative value, standardized accuracy and standardized wrong diagnostic rate of the method were 90.91%, 82.22%, 83.64%, 90.04%, 86.57% and 13.43%, respectively. The cutoff value of serum TTF-1 in healthy normal adults was 36.39, with a ROC value of 0.95. Serum TTF-1 PU was significantly higher in patients with lung adenocarcinoma, squamous cell lung carcinoma and thyroid carcinoma than in healthy adults and patients with benign lung diseases and non-thyroid carcinoma (P=0.000). Serum TTF-1 PU was similar in lung adenocarcinoma, squamous cell lung carcinoma, small cell lung carcinoma, large cell lung carcinoma and thyroid carcinoma (P=0.744, 0.677, and 0.333, respectively). Serum TTF-1 PU was greater than the PU in the corresponding homogenate of lung adenocarcinoma, squamous cell lung carcinoma, small cell lung carcinoma, large cell lung carcinoma and thyroid carcinoma (P=0.000). Serum and homogenate TTF-1 PU was correlated to TNM stage of lung cancer patients (P=0.000) but not to gender, tumor types, differentiation or lymph node metastasis.

CONCLUSIONSLung adenocarcinoma, squamous cell lung carcinoma and thyroid carcinoma are suspected when serum TTF-1 PU is higher than 36.39. Serum TTF-1 is not helpful in differentiating the types of lung carcinomas and thyroid carcinoma. After exclusion of thyroid carcinoma, detection of serum TTF-1 can be helpful in the diagnosis of lung cancer. In different lung carcinomas and thyroid carcinomas, the serum TTF-1 is higher than the corresponding homogenate TTF-1 level. Serum TTF-1 increases with the progression of TNM stage of lung carcinoma.

Case-Control Studies ; Humans ; Lung Neoplasms ; blood ; classification ; diagnosis ; Nuclear Proteins ; blood ; Predictive Value of Tests ; Sensitivity and Specificity ; Thyroid Nuclear Factor 1 ; Transcription Factors ; blood