Objective: To observe the expression of CD133 in the clear cell renal cell carcinoma (RCC) and to investigate the related drug resistance. Methods: Flow cytometry and immunohistochemical method were used to examine the expression of CD133 in metastatic RCC cell line RCC05-TXJ, low metastatic RCC cell line RCC05-ZYJ and two clinical non-metastatic RCC primary cultures isolated from a male and a female patient. The four cell lines were treated with IFN-α and 5-FU and the viability of cells were examined by MTT assay before and after treatment. Results: Flow cytometry showed that RCC05-TXJ and RCC05-ZYJ expressed CD133. The in situ carcinomas of male and female patients hardly expressed CD133. Immunohistochemistry examination showed that the local membrane of RCC05-TXJ and RCC05-ZYJ cells expressed CD133. RCC cells of female and male patients hardly expressed CD133. RCC05-TXJ and RCC05-ZYJ cells exposed to IFN-α or 5-FU showed a rebound of survival 24 h after withdrawal of drugs. The survival rates of RCC cells of male and female patients kept at a low level after withdrawal of drugs. Conclusion: There are a small number of CD133+ RCC cells in RCC patients, with a property similar to tumor stem cells, which might be one of the important factors influencing the immunotherapy and chemotherapy of tumors.

Objective: To establish clear cell renal cell carcinoma (ccRCC) cell lines from clinical ccRCC specimens of Han nationality in China and to characterize the biological features. Methods: From 2005 to 2007, fresh surgical samples of ccRCC were obtained from 43 patients; the samples included primary tumor in situ, osseous metastasis, lymph node metastasis, and cancerous embolus. The samples were cultured in vitro using explant-culture method within 30-60 min after surgery. Analysis on cell growth and colony-forming efficiency was recorded for the lines which were passaged for over 50 generations. Chromosome examination, pathological examination and tumorigenesis in NOD-SCID mice were used to determine their malignancy. Flow cytometry was used to determine expression of CA9 and CD133. Results: Most of the primary cells could only be passaged for less than 5 generations; 5 lines could be serially passaged for over 5 passages, 3 lines for over 10 passages, and only 2 lines could be stably passaged. One line,named RCC05-TXJ, was from osseous metastatic ccRCC and had been serially passaged for 110 generations in 21 months; the average doubling time was 19.2 h,average chromosome number was 75,and colony forming efficiency was 41%. Another line,named RCC05-ZYJ,was from primary ccRCC specimen and had been serially passaged for 160 generations in 18 months; the average doubling time was 16.5 h,average chromosome number was 55,and the colony forming efficiency was 37%. Immunohistological analysis demonstrated that both lines expressed CA9 and CD133. Flow cytometry analysis found that expression levels of CA9 and CD133 increased with the passages. Both RCC05-ZYJ and RCC05-TXJ lines were able to form tumor and to metastasize in NOD-SCID mice; however, their metastatic ability was obviously different. Conclusion: We have established 2 ccRCC cell lines with different metastatic potentials from the clinical ccRCC specimens of Han nationality in China. The ratio of tumor stem cells increases with the passages.

Objective To investigate the role of extracellular signal-regulated kinase(ERK)signal pathway in the spinal cord in the development of chronic morphine tolerance.Methods Thirty healthy male SD rats weighing 300-350 g in which intrathecal(IT)catheters were successfully implanted without complication were randomly divided into 3 groups(n = 10 each): group Ⅰ control(group C);group Ⅱ morphine tolerance(group M)and group Ⅲ morphine tolerance + PD98059(ERK upstream kinase MEK inhibitor)(group P).Morphine tolerance was induced with IT morphine 10 μg twice a day for 7 consecutive days.In group P PD98059 10 μg was injected IT at 30 min before each morphine administration.Tail flick latency(TFL)(the time between the onset of heat stimulus and voluntary tail withdrawal)was measured once a day at 30 min after first IT morphine administration and at 1 day after last IT morphine.Maximum analgesic effect(MPE)was calculated.MPE =(TFL after IT morphine - baseline TFL)/(12 - baseline TFL)× 100%.The animals were sacrificed after last TFL measurement.The L3-5 segment of the spinal cord was isolated for determination of the expression of μ-receptor and phosphorylated ERK 1/2(p-ERK1/2)by Western blot analysis and fluoroimmunoassay.Results Morphine tolerance was induced in group M and M + P.MPE was higher in group P than in group M.The expression of μ-receptor in spinal dorsal horn was significantly lower while the p-ERK1/2 expression was higher in group M than in group C.IT PD98059 significantly up-regulated μ-receptor expression and down-regulated p-ERK expression in group P as compared with group M.Conclusion ERK signal pathway is involved in the development of chronic morphine tolerance in rats.

Objective To establish a HPLC method to determine the related substances of vitamin B6 for injection.Methods HPLC method was adopted with DiamonsilTM C18,5μm 250mm×4.6mm;the mobile phrase was length was 291nm.Results The calibration curve was linear in the concentration range of 600～1400μg/ml(r=0.9998).Conclusion The method is simple,rapid,accurate and reliable.As the supplement method of active pharmacopoeia,it can effectively control the quality of the product.

Objective To investigate the clinical and endoscopic features of ischemic colitis (IC),to provide a reference for clinical diagnosis and treatment.Methods A retrospective analysis of 62 inpatients who were diag-nosed as IC was performed.The patients'clinical data,clinical manifestations,complications,laboratory tests,imaging tests,colonoscopy,pathology,diagnosis,treatment,follow -up and other information were analyzed.Results Of 62 patients,there were 1 7 males and 45 females,mean age was (66.1 ±1 0.7)years old,more than 60 years of age there were 47 cases (75.81 %),of which 38 cases (61 .29%)showed the presence of one or more underlying disea-ses.All patients had acute onset,57 cases (91 .94%)had abdominal pain,bloody stools occurred in 56 cases (90.32%).Colonoscopy revealed 60 cases as a transient type,2 cases of stenosis,no gangrene type,lesions were mainly left colon.All patients were given fasting,anti -infection,microcirculation improvement and intravenous nutri-tional support,with no serious complications or transfer surgery occurred,colonoscopy was reviewed 4 -8weeks, 62 cases were healing or had improvement,2 patients in the 3 -year observation period was readmitted attack,the recurrence rate was 3.23%.Conclusion Ischemic colitis is more common in older people,to strengthen the aware-ness and vigilance of the disease,early diagnosis based on colonoscopy as a primary diagnostic modality and prompt treatment has positive significance.

Objective To improve the knowledge of CT appearances of spondylolysis of the lumbar spine.Methods 32 patients with lumbar spinal spondylalysis were examed by Elscint 2400 CT.CT appearances were analyzed.Results The CT feature of spondylolysis was loop-broken sign.Spondylolisthesis was present in 25 cases,the CT mainfestations included the increased anterior pasterior diameter of spinal canal,wich sign,deformity of vertebral canal and pseudobulging disk.Other features included asymmetrical zygapophysial joints.Conclusion “loop-broken sign” is a relatively specific CT appearance of spondylolysis of the lumbar spine,which is significant in diagnosting spondylolysis without spondylolisthesis.

By comparing the differences of " Decision of the Standing Committee of the National People's Congress on the prohibition against narcotic drugs " and "Law of the People's Republic of China on Narcotics Control " and their implications for harm reduction,the article analyzed the ethical issues existing in applying harm reduction policy in HIV/AIDS prevention and control,and held that the new policy to promote the methadone use would play a major role for control of drug abuse and AIDS.

Objective The prestin,a motor protein responsible for outer hair cell(OHC)electromotility,is expressed on the OHC surface.Previous experiments revealed that OHC electromotility and its associated nonlinear capacitance was mainly located at the OHC lateral wall and was absent at the apical cuticular plate and the basal nucleus pole.Immunofluorescent staining for prestin failed to demonstrate the prestin expression at the OHC basal ends in whole-mount preparation of the organ of Corti.The aim of this study is to investigate the distribution of prestin at OHC.Methods In this experiment,the localization of prestin protein in single dissociated OHCs from cochlea of normal mouse,rat and guinea pig,were examined by immunofluorescent staining and confocal microscopy.Results We found that prestin was uniformly expressed on the OHC basolateral surface,including its basal pole.No staining was observed on the cuticular plate and stereocilia.The OHC lateral wall had a trilaminate organization and was composed of the plasma membrane,cortical lattice,and subsurface cisternae.By with co-staining with a membrane marker di-8-ANEPPS,prestin-labeling was found locating at the outer layer of the OHC lateral wall.Further separating the plasma membrane from the underlying subsurface cisternae,using a hypotonic extracellular solution,prestin-labeling was shown locating at the plasma membrane instead of the subsurface cisternae.Conclusion The data revealed that prestin is expressed in the plasma membrane on the whole OHC basolateral surface.

AIM: To establish an HPLC-MS for determining seven bioactive components,including vanillin ferulaic acid,ligustrazine,senkyunolid I,H,and A,and ligustilide in Ligusticum chuanxiong Hort. . METHODS: The bioactive components were extracted by ultrasonic method,and then were separated by reverse HPLC,and finally were determined by the multiple reaction mode of MS. RESULTS: The detected limits of the seven bioactive components such as vanillin were 40,20,80,20,10,60 and 50 ng/mL,respectively. Their linear range were 1450,4-490,1-625,2-360,2-700,1-244 and 21-210 ?g/mL,respectively. The average contents of the above components in Ligusticum chuanxiong Hort. were 0. 12,0. 25,0. 41,0. 99,0. 30,8. 96 and 11. 25 mg/g,respectively. CONCLUSION: The proposed method has the characteristic of simplicity,rapidty and high sensitivity. It could be used in analyzing the whole components of Ligusticum chuanxiong Hort. .

The diameter of the mitral orifice was measured in 122 cases of rheumatic mitral stenosis when commissurotomy was performed. In 99 cases(81.25%) with a history shorter than 10 years, the diameters ranged from 0.3 to 2 cm; it was larger than 1.1 cm. in 18 cases and smaller than 0.8 cm. in 68 cases.After analyzing the interrelationship between the diameter of the mitral orifice and the clinical manifestations, ECG, the postoperative complications, and the pathological findings, it was found that the size of the mitral orifice was not always in direct proportion with either the duration of the illness or the severity of cardiac enlargement. In those patients with a mitral orifice smaller than 0.8 cm., the following criteria were often present:1. Cardiac function above Ⅲ degree.2. Repeated attacks of hemoptysis.3. Presystolic contraction on ECG.4. Postoperative complication of auricular fibrillation.5. Thrombosis on the cardiac walls detected pathologically.