Objective To investigate the possibility of affecting transgenic rescue for Wilson disease using the human ATP7B transgenic LEC rat.Methods The 7.1kb transgene constructed with human ATP7B cDNA and chicken ?-actin promoter was introduced into the fertilized oocytes of LEC rats, an animal model of Wilson disease, by microinjection. The expressions of human ATP7B protein in the transgenic rats were detected by Western blot. The plasma AST and ALT activities, and the total bilirubin levels in transgenic rats were measured continuously from 6 to 16 weeks using non-transgenic rats and LEA rat as controls. The pathological and histochemistry changes in the liver of the transgenic rats at 13 weeks were analyzed. Results The intact and correct product derived from human ATP7B was confirmed in the liver of transgenic rats. At the age around 12 weeks, the plasma AST and ALT activities, and the total bilirubin levels in transgenic rats were significantly decreased, while the inflammatory reation in the liver of transgenic rats was much mild as compared with that of non-transgenic rats, and the granules of stained copper were less in the hepatocytes of transgenic rats. By the age of 16 weeks, the transgenic rats were phenotypically normal, and the survival rate was 100%. These data showed that the LEC rats were successfully rescued from fulminant hepatitis after introducing of human ATP7B gene. Conclusion The hepatitis in Wilson disease is directly related to the toxicity of excessive accumulated copper, which attributed to the functional deficiency of the ATP7B. Gene transfer probably is the effective method for the therapy of Wilson disease.

Objective To investigate the possibility of affecting transgenic therapy for the hepatic cirrhosis and hepatoma in Wilson disease by human ATP7B cDNA. Methods The 7.1*!kb transgene consisting of human ATP7B cDNA and chiken ?-actin promoter was introduced into the LEC rats which is an animal model of Wilson disease by microinjection.The plasma AST and ALT activities in transgenic rats were measured continuously from weeks 17 to 30 using non-transgenic and LEA rats as controls.The histological and histochemistry changes of liver in the transgenic rats at 30 and 60 weeks of age were examined. Results The plasma AST and ALT activities in transgenic rats were kept at the relatvie lower levels from 17 to 60 weeks of age, as compared to the age-matched non-transgenic rats.By the age of 60 weeks,none of the transgenic males developed cholangiofibrosis or hepatoma,whereas all of the non-transgenic rasts had severe cholangiofibrosis at the age of 30 weeks and one male rat had hepatoma at 60 weeks.The transgenic rats were phenotypically normal,and the survival rate was 95.7%.In addition,the distribution and the numbers of the granules of stained copper in the hepatocytes of the transgenic rats did not show any significant difference between 30 and 60 weeks.Conclusion The human ATP7B successfully delayed the onset of hepatic cirrhosis,and suppressed the development of hepatoma in the LEC rats by gene transfer.The hepatic cirrhosis and hepatoma in Wilson disease may be not directly related to the copper accumulation.

Objective To explore the correlation of pathological changes and imaging through a control study on pathology and imaging of 4 cases of senile dementia of the Alzheimer type (SDAT).Methods By HE and Gallyas silver dyeing,four immunohistochemical methods were carried out The loss of neurons cells of all lobes of cerebral cortex,cingulate gyrus,amygdaloid nucleus,hippocampal gyrus,parahippocampal gyrus,meynert basal nucleus,substantia nigra,locus ceruleus,dorsal nucleus of vagus nerve,senile plaques (SP) and the changes of neurofibrillary tangles (NFT) were observed in detail The control study of pathology and imaging was carried out by comparison of CT in different phases before and after death of the patient.Results The degeneration of temporal lobe was marked: (1) The degeneration of middle frontal gyrus of cortex of frontal lobe was the most evident; (2) The degeneration of long gyrus was more evident than that of the short gyrus as the cortex of insular lobe was concerned; (3) The degeneration posterior part of cingulated gyrus was more evident than that of the anterior part.Conclusion The pathological findings of SDAT mentioned above are basically in conformity with the changes of imaging.

Objective To evaluate the clinical efficacy and toxicity of rituximab combined with CHOP (R-CHOP) in the treatment of relapsed or refractory diffuse large B cell lymphoma (DLBCL).Methods 30 patients were enrolled.All patients,pathologically confirmed to be CD20 positive DLBCL (all in stages Ⅲ-Ⅳ ), were relapsed or refractory after received 2-6 cycles of chemotherapy. Then all of them received R-CHOP schedule for 4 to 6 cycles, each cycle was 21 days. Clinical data before and after R-CHOP were collected.A retrospective analysis of the R-CHOP therapy,either compared to the literature or self-control was performed to evaluate the efficacy and toxicity. Results All the 30 patients were evaluable, induding 15 cases were complete remission (CR), 10 cases were partial remission (PR), 3 cases were stable disease (SD),and 2 cases were progressed disease (PD).The CR rate was 50.0 ％ (15/30),the total response rate (RR) was 83.3 ％ (25/30).All patients were well tolerated to the therapy.Only 3 cases were Ⅱ degree neutropenia,1 case was Ⅰ degree thrombocytopenia, and 2 cases suffered nausea and other mild gastrointestinal discomfort. Conclusions R-CHOP regimen could also achieve good response for relapsed or refractory DLBCL significantly.The common adverse effects of rituximab were mild.All the patients were well tolerated.

Objective To analyze the expression profile of specific miRNA of gastric cancer tissues.Methods miRNA were isolated from five paired gastric cancer tissues and corresponding paracancerous normal tissues.To identify the profile of gastric cancer,miRNA hybridization and cluster analysis were processed by significance analysis of microarrays (SAM,version 2.1) and cluster software.The relative expression of hsa-miR-30c,hsa-miR-196a and hsa-miR-193b were validated by fluorescence Real-Time PCR using taqman probe.Further hsa-miR-30c taget gene was predictived by online software.Results Revealed by miRNA microarray detection,there were 16 miRNA with differential expression in gastric cancer tissues and para-cancerous normal tissues,among which HS_100,hsa-miR-27a,hsa-miR-214*,solexa-555-1991 and hsa-miR-196 were significantly up-regulated,and hsa-miR-502-3p,hsa-miR-29c*,hsa-miR-64,hsa-miR-193b,hsa-miR-551b,hsa-miR-30c,hsa-miR-582-5p,hsa-miR-625*,hsa-miR-660,hsa-miR-363 and hsa-miR-486-5p were down-regulation.The relative expression of hsa-miR-30c,hsa-miR-196a and hsa-miR-193b was in high concordance with microarray.Conclusion There is specific miRNA expression profile in gastric cancer tissues,among which differentially-expressed miRNAs may be involved in the regulation of gastric cancer progress and serve as new potential biomarkers.

Objective To study the clinicopathology characteristics of chronic lymphocytic thyroiditis (Hashimoto's thymiditis, HT) with occult papillary thyroid carcinoma (occult PTC) in order to improve its diagnostic and therapeutic accuracy. Methods A restrospective analysis was done on the clinical materials of 28 cases of HT with occult PTC from July 1999 to July 2005. All cases were confirmed by operation and pathologic biopsy. Clinical and gross findings were collected. All HE slides were reexamined and immunostains for CK19, galectin-3,and bcl-2 were performed (Envision method). Results In total 189 cases of chronic lymphocytic thyroiditis,28cases (14.8%,28/189)had coexistent occult papillary thyroid carcinoma. There were 19 females and 9 males with median age of 36.7 years old. Normal TSH presented in 11cases (39.3%), high in 8 cases(28.6%), and low in 9cases (32.1%). All tumor diameter was counted for <0.8 cm, 16 cases (57.1%) tumor diameter 0.2～0.5 cm, 12 cases (42.9%) >0.5 cm. Coarse calcification was seen in 6 cases(21.4%) in color ultrasonic exam and CT scans. Follow-up data showed that 28 patients were all alive with no evidence of recurrence or metastasis for 2 to 7 years by December 2007. Conclusion There are no special clinical characteristics in coexistent HT with occult PTC.Coarse calcification in HT in the group of middle-aged women increase the likelihood of the diagnosis. But the diagnosis depends on pathology. Because of the high incidence of occult PTC in HT population, it would be necessary to keep an eye on this particular type of thyroid carcinoma, and multiple sampling in suspected area of HT specimen is advised in the hope not to miss any small tumor in clinical practice.

Objective To investigate the expression of ionic channels of the rat bone marrow mesenchymal stem cells(BMSCs) and provide the basis for the change of ionic channels in the process of differentiation of stem cells.Methods The full marrow from Wistar rats was isolated by Percoll centrifugation and passaged repeatedly.The membrane currents of BMSCs were recorded with the whole-cell patch-clamp technique.The current amplitude of pathway,peal value and time course of activation and deactivation,selectivity of ions pathway,I-V curve were recorded.The currents were identified and analyzed by using various channel blockers.Results The cells began to stick in 24 h and arranged in whirlpool shape after passenger culture.They were positive for CD71,CD44,and negative for CD34,CD45 by flow cytometry,which improved that the cultured cells were BMSCs.The functional ion channels were characterized in cultured rat BMSCs with whole-cell patch clamp and reverse transcription polymerase chain reaction(RT-PCR) techniques.Three types of outward currents were found in BMSCs,including a delayed rectifier K+ current(IKDR),in addition,tetrodotoxin-sensitive sodium current(INa.TTX) and nifedipine-sensitive L-type Ca2+ current(ICa.L) were detected in 24% and 25% BMSCs.Moreover,RT-PCR result revealed the molecular evidence of mRNA for the functional ionic currents,including SCN5A,Kv4.3 and CACNA1C.Conclusion The currents are consistent with the characteristics of excitable cell membrane.

Objective To investigate the therapeutic effect of total mesorectal excision(TME) for low rectal cancer.Methods One hundred nineteen patients with rectal cancer,located on an average within 8 cm of the anal verge,were included in the study.Fifty-four patients underwent traditional operation,and sixty-five patients underwent total mesorectal excision.Operation time,loss of blood at operation and local recurrence rates were compared between the two groups.Results The average operation time and blood loss were 118 minutes,and 100mL respectively in TME group,and they were 182 minutes,and 340 ml respectively in traditional operation group.There were significant differences between them(all P0.05).Conclusions In TME group,operation time was shorter,operative blood loss was less,and local recurrence rate was lower.TME should be applicated for patients with low rectal cancer.

OBJECTIVE： To develop a HPLC method for the determination of the contents of lidocaine and prilocaine in compound lidocaine cream as a quality control means.METHODS： Lidocaine and prilocaine in compound lidocaine cream were determined by high-performance liquid chromatography on C18 column with the detection wavelength at 254nm.The mobile phase was 0.5％ ammonium dihydrogen phosphate(pH=7)-methanol(20∶ 80).RESULTS： The calibration curves of both lidocaine and prilocaine were linear within the concentration range of 130～250μ g/ml(r=0.9 996).The recovery rates of lidocaine and prilocaine were 99.05％ and 99.27％ respectively， RSDs were 0.67％ and 1.15％ ， intra-day RSDs 0.81％ and 1.45％ ， inter-day RSDs 0.55％ and 0.63％ respectively.CONCLUSION： The method was sensitive， stable and accurate.It can be used to determine and control the quality of compound lidocaine cream.

To observe Antiarrhythmic effects of L-dicentrine．Methods using experimented model of arrhythmials．Results: L-dicentrine iv l0 mg/kg could convert BacRinduccd arrhythmia into sinus rhythm in rats,and could significantly increase the tolerant dose of aconine to produce Ventricular fibrillation (VF) and Cardiac ［rrest (ca) in rats． The drug could also decrease the incidence of VF and CP by cardiac rats and by chloroform in mice． L-dicelltrine had protective effects on ouabain ind uced　Arrhythmias in guined pigs and on adrenalice induced arrhythmias in rabbit s ． The drug might also inhibit the arrhymias induced by stimulation ofhypothaplu s． Conclusion: L-dicenirine have significant antiarrhythmic properties．