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.

Object To investigate the effect of PPARαactivation on PPARγ-induced fatty liver in the mouse. Methods Wild type mice ( C57BL/6) aged 4 to 5 weeks were used as animal models.All mice were divided into four groups.The mice in the first group were fed with chow diet.The mice in the second group were fed with a diet containing 0.125%Wy-14,643, an agonist of PPARa, for 8 days.The mice in the third group were injected with Ad/PPARγvia tail vein for 5 day.The mice in the fourth group were firstly fed with Wy-14,643 diet for 3 days and then injected with Ad/PPARγvia tail vein for another 5 day.Mouse livers were collected and photographed.The effect of PPARαactivation on PPARγ-induced fatty liver was observed by H&E and Oil red O staining.Results Compared with the controls, wild-type mice treated with Wy-14,643 for 8 days exhibited marked hypertrophy of hepatocytes with increased cytoplasmic eosinophil-ia and proliferation of peroxisomes.The liver size was significantly increased in the wild-type mice treated with Ad/PPARγfor 5 days, and over-expression of PPARγstrongly induced hepatic steatosis.Importantly, the wild-type mice pretreated with Wy-14,643 for 3 days and then given Ad/PPARγinjection exhibited dramatically the increase of liver size, which might be due to the dual function of PPARa and PPARγ.Compared with the Ad/PPARγgroup, the Wy-14,643 pretreat-ment group showed a reduced hepatic steatosis.Conclusions Activation of PPARαby Wy-14,643 effectively improves PPARγ-stimulated hepatic steatosis, which provides a novel target for prevention and therapy of fatty liver.

Background and purpose:Most breast cancer cell lines were derived from malignant pleural fluid or other tissues that metastasized from breast carcinoma.Breast carcinogenesis is under represented due to the significant distinction of biological characteristics between cancer cell lines and primary breast cancers.Few primary breast tumors spontaneously developed into immortal cell lines.Overexpression of human telomerase reverse transcriptase(hTERT) in normal epithelial cells led to immortalization.We aimed to establish immortalized cultures derived from primary breast cancer by introducing hTERT and determine the gene profile of hTERT-transfected cultures.Methods:hTERT was introduced into breast tumor cultures with a puromycin-resistant retroviral construct,pBABE-hTERT-puro.Real-Time RT-PCR was used to analyze the expression level of hTERT after transfection.The upregulated gene profile was determined with microarray techniques.Results:hTERT was stably over expressed,at least 1 400-fold in hTERT-transfected cells compared to pBABE-puro transfected cultures.These ectopic over-expressions led to immortalization of transfected cultures,which could undergo at least 40-50 passages.Microarray analysis further confirmed the over-expression of hTERT in transfected cultures and demonstrated that 22 genes were up-regulated due to over-expression of hTERT.No down-regulated genes were observed cross all cultures.Conclusions:Over-expression of hTERT led to immortalization of breast tumor primary cultures and up-regulated certain genes.

Objective To investigate the effects of Tanshinone Ⅱ A (extracted from Chinese herb medicine Salvia miltiorrhiza Bge.) on ventricular arrhythmias of rabbit hearts induced by ischemia in order to illuminate its mechanism of anti-arrhythmia.Methods Thirty rabbits were randomly (random number)divided into normal group,ischemic group and Tanshinone Ⅱ A group.Model of wedge shaped mass of rabbit left ventricular myocardium with coronary perfusion was prepared,and then by using floating glassy microelectrode,the trans-mural ECG,QT interval,the trans-mural dispersion of re-polarization (TDR) and trans-membrane action potentials from both endocardium and epicardium were simultaneously and wholly recorded.The incidence of ventricular arrhythmia in myocardium was observed after ischemia for thirty min.Results Under the condition of acute ischemia,compared with normal group,the incidence of ventricular arrhythmia and TDR were significantly increased in ischemia group (P ＜ 0.01),while incidence of ventricular arrhythmia and TDR were significantly reduced in tanshinone ⅡA group compared with ischemia group (P ＜ 0.05).The incidences of ventricular arrhythmia in normal,ischemia and Tanshinone Ⅱ A groups were 0/10,9/10 and 2/10 respectively.Conclusions Tanshinone Ⅱ A prevents ventricular arrhythmia and reduces TDR significantly in ischemic rabbit hearts.

Hernia, Inguinal ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Peritoneal Dialysis, Continuous Ambulatory ; Peritoneum ; diagnostic imaging ; Tomography, X-Ray Computed ; methods

0.05).Conclusion MRI is a highly accurate diagnostic tool for detecting tears of the medial and lateral menisci.At present,there is no evidence to ascertain that higher magnetic field strength improves discriminatory power for meniscal tears.

In order to effectively remove the invalid impurities in Tongan injection, optimize the optimal parameters of the impurity removal technology of liquid mixing process, in this paper, taking Tongan injection as the research object, with the contents of celandine alkali, and sinomenine, solids reduction efficiency, and related substances inspection as the evaluation indexes, the removal of impurities and related substances by the combined process of refrigeration, coction and activated carbon adsorption were investigated, the feasibility of the impurity removal method was definited and the process parameters were optimized. The optimized process parameters were as follows: refrigerated for 36 h, boiled for 15 min, activated carbon dosage of 0.3%, temperature 100 degrees C, adsorption time 10 min. It can effectively remove the tannin, and other impurities, thus ensure the quality and safety of products.
Adsorption ; Charcoal ; chemistry ; Drug Compounding ; instrumentation ; methods ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Quality Control

Objective To study the genotype distribution of insulin receptor substrate-2(IRS-2)gene 1057G/A polymorphism in Han population from Southwest China,and to explore its association with the metabolism of glucose and lipids,insulin resistance and islet?-cell function in type 2 diabetic patients and subjects with impaired glucose tolerance(IGT).Methods A total of 929 Hans[462 patients with type 2 diabetes(DM group) and 164 subjects with IGT(IGT group)and 303 normal controls(NC group)]from Chongqing and nearby regions were screened for 1057G/A polymorphism of IRS-2 gene by PCR-RFLP assay.Body mass index(BMI),plasma glucose,serum insulin and lipid profile,high-sensitive C-reactive protein(hsCRP)and non-esterified fatty acid were measured.Homeostasis model assessment of insulin resistance(HOMA-IR)and disposition index(DI)were used to estimate insulin resistance and?-cell function respectively.Results In DM group,A allele frequency was significantly lower than that in NC group(0.326 vs 0.388,X~2=6.19,P=0.01).Compared with NC group,AA genotype frequeney was lower and GG genotype frequeney was higher in DM group(0.104 vs 0.135 and 0.452 vs 0.360 respectively,X~2=6.80,P

Objective In order to explore the effects of rehabilitation training and acupuncture on the neural function deficit and motor function in patients with ischemic streke.Methods Eighty patients with ischemic stroke were randomly divided into rehabilitation and acupuncture groups.According to different recovery stages,the rehabilitation group received purposeful rehabilitation training for 28 days while the acupuncture group received scalp needling combined with body acuptmcture.The neural deficit scores(NDS)and motor fimction comprehensive assessment(FCA) were evaluated at the recruitment(M_0),the 28~(th)(M_1)and 56~(th)(M_2)days after treatment.Results No significant difierences were found in the NDS and motor FCA between the rehabilitation and the acurluncture groups at the recruitment.The significant differences appeared at the 28~(th) and the 56~(th) days comparing with baseline assessments in each group.There was no significant difference in the changes of NDS and motor FCA between the two groups at 28~(th) day,however,the NDS and motor FCA in rehabilitation group revealed better outcome than the acupuncture group at the 56~(th) day.Conclusion The study indicated that both rehabilitation training and acupuncture could improve the neural function and enhance the motor function in patients with ischemic stroke.

Full-length NS5A gene of the hepatitis C virus was amplified by PCR using plasmid pBAC25 containing HCV nonstructural gene as template. The amplified fragment (about 1.34 kb) was cloned into plasmid pQE32, and the recombinant plasmid pQENS5A was expressed in JM109 strain. The NS5A protein was purified by NiSO4 metal chelating resin, and characterized by Western-blot. Its antigenecity was determined by ELISA. The positive detection rate of anti-NS5A was 75% (69/92) in ninety-two clinic sera. The positive rate of anti-NS5A was 82.5% (33/40) in fourty positive standand sera, and the negative rate of anti-NS5A was 100% (40/40) in fourty negative standand sera. The results showed that the Full-length NS5A proteinn had the higher sensitivity and specificity in the detection of HCV antibody in sera, we suggested that NS5A protein was a useful antigen for blood screening.