Objective Multidrug-resistance (MDR) is considered to be the major obstacle for cancer chemotherapy.In order to study tumor MDR in vitro, we designed this study to establish human multidrug-resistant Bladder cancer pumc-91/ADM cell line and investigate its biological characteristics. Methods MDR cell line (Pumc-91/ADM) was induced by wise selection on exposure to increasing dose of Adriamycin (ADM).Cell growth was measured and multidrug resistance to multi-anticancer agents was evaluated by MTT Assay.Flow cytometry was performed to determine cell cycle and the ADM concentration of cell line. The expression of MDR-related genes were determined with reverse polymerase chain reaction (RT-PCR). Results Compared to Pumc-91, the Pumc-91/ADM cell had a prolonged doubling time. The number of cells in S-phase was decreased in Pumc-91/ADM while those in G1 and G2 phase increased. The Pumc-91/ADM cell was 10 times more resistant to ADM than the Pumc-91 parent. The Pumc-91/ADM cell exhibited cross-resistance to methotrexate, vincristine, cisplatin, epirubicin. RT-PCR showed that mRNA expression of GST was significantly increased in Pumc-91/ADM. Conclusion Pumc-91/ADM is human multidrug-resistant, and it offers a model with MDR phenotype for the study of MDR in human bladder cancer.

Objective To find a indicator in urine to assist diagnosis of cerebral infarction,we investigated the changes of urine ferritin in patients with cerebral infarction.Methods Collected serum from 30 healthy volunteers and 53 patients with cerebral infarction(CI),with ratio of males to females 19∶11 vs.30∶ 23,and average age was (56 ± 11) vs.(63 ± 11) respectively.Collected urine from 39 heathy volunteers and 33 patients with CI,with ratio of males to females 25∶14 vs.20∶ 13,and average age was (57 ± 10) vs.(64 ± 11) respectively.Ferritin concentration in all samples was detected with the chemical luminescence method,and the urine ferritin/urine cretinin (uFer/uCr) ratio was calculated.Differences of serum ferritin and uFer/uCr between healthy volunteers and CI patients were analyzed with the statistical software under Wilcoxon test.Urinary proteins were extracted with absolute ethanol.The extracted ferritin was displayed with Western blot for light chains and heavy chains to investigate the difference between healthy volunteers and CI patients.Results uFer/uCr was higher than that from healthy volunteers (U =292.00,P ＜0.05),and the median was (14.86 vs.6.22) μg/g.Moreover,after extraction of urinary proteins with alcohol,both light chains and heavy chains of ferritin in urine from CI patients were much more obvious than that from healthy volunteers.Conclusion The uFer/uCr ratio could provide a new proof to assist the diagnosis of cerebral infarction.

Objective To construct a prokaryotic expression system of human ferritin L which could be overexpressed in E.coli, and then prepare quality control materials of rFL and evaluate the homogeneity, stability, precision and application value of the rFL.Methods RT-PCR was used to clone ferritin L gene with total RNA from peripheral blood.Ferritin L gene was inserted into plasmid pGM-T to construct subclone plasmid pGM-T/ferritin L, which was identified by sequencing.The recombinant plasmid pET-30a/ferritin L was transformed into E.coli BL21 (DE3) and efficiently expressed under IPTG induction.rFL was identified by SDS-PAGE, and its antigenicity was examined by WB.The concentration of rFL was measured by ACCESS immunoassay system.Fifteen control samples were randomly selected using random number table.The homogeneity and stability of rFL were evaluated by one-way analysis of variance (ANOVA) and a linear regression model, respectively.The uncertainties of homogeneity, stability and the precision were evaluated.Results A prokaryotic expression system of ferritin L was successfully constructed.Sequence analysis showed that the ferritin L gene inserted was identical with the one from GenBank(NM_000146.3).The PCR products were 527 bp long, in complete agreement with length of ferritin L gene.The molecular weight of rFL highly expressed in E.coli induced by IPTG was 19 000 Da.The WB analysis indicated that this rFL protein had good antigenicity.The concentration of rFL(1 000 times dilution) measured by ACCESS immunoassay system was (1115.84±38.38) ng/ml.Homogeneity evaluation of low-concentration QC sample and high-concentration QC sample of rFL showed that there were no statistical significances in the within-groups and between-groups (for high-concentration F=2.336, P>0.05 and for low-concentration F=0.730, P>0.05).A linear regression based on the stability test indicated that there was no statistically significant trend of instability in five and a half months (F=1.755, P>0.05). Precision analysis showed the within-run CV, the between-run CV, the between-day CV and the total CV of high-concentration QC sample were 2.06%, 2.12%, 0% and 2.96% respectively; the within-run CV, the between-run CV, the between-day CV and the total CV of low-concentration QC sample 2.03%, 2.08%, 0% and 2.90%, respectively.Conclusion This rFL for quality control materials with independent intellectual property rights could meet the clinical requirement of homogeneity, stability and precision, and could provide raw materials for preparation of mixed ferritin for quality control.

Objective To investigate the influence of illumination changes on the drosophila circadian rhythm and 24 h sleep time.Methods We collect the unmated fruit flies (a total of 128,single-sex) under CO2 anesthesia within 8 hours of eclosion.And put them into the group of normal light group,day and night reversed group,24 h light group and 24 h dark group with the simple random sampling method,male and female with 4 single-sex groups(n=32).Change the illumination conditions of the living environment of the fruit flies and use Drosophila Activity Monitoring System (DAMS) and Data Acquisition System (DAS),to observe and compare the changes of drosophila circadian rhythm and 24 h sleep time between the different illumination conditions.Results Compared with the normal light group,drosophila circadian rhythm of the day and night reversed group was significantly reversed,the number of fruit flies' activities of 24 h light group was increased,and circadian rhythm was decline,drosophila circadian rhythm of the 24 h dark group is still there,but frequent activity appears during the day,and the 19 : 00 activity peak disappears.Conclusion The drosophila circadian rhythm and 24 h sleep time changes with illumination conditions change.

Objective To assess the efficacy and safety of esomeprazole in long-term or intermittent treatment of gastroesophageal reflux disease (GERD). Methods Twenty-eight patients with GERD who accepted esomeprazole 20 mg bid for 2 weeks were further divided into long-term treatment group and intermittent treatment group according to the protocol of therapy. Patients in long-term treatment group were received minimum dose that was needed to relief the symptoms for more than 6 months, whereas those in intermittent treatment group were received esomeprazole 20 mg qd until the symptoms completely disappeared, if symptoms relapsed the patients were treated again.The dosage, recurrence of symptoms and the side effects were compared between two groups. ResultsThirteen patients in long-term treatment group were treated for 7-44 months (20 mg daily in 7,twice daily in 5 and every other day in 1). While 15 patients in intermittent treatment group had a good relief of the symptoms. No adverse reactions was found in two groups. The follow-up study of 10-57 months in intermittent treatment group revcaled that the longer the treatment maintained,the longer the symptoms relieved (r=0. 447, P= 0. 008). Conclusion It is safe for esomeprazole in relieving symptoms of patients with GERD by long time or intermittent use.

Objective To study the clinical law and imaging characteristics of intracranial primary malignant lymphoma.Methods The clinical law and imaging finding in 27 cases of intracranial primary malignant lymphoma confirmed by pathology were analysed CT scan was performed in all cases.MRI examination were carried out in 20 cases simultaneously.Results The first symptom in most cases was headache,and other symptoms were epileopsy,dizziness,hemiplegia,etc.This lesion was mostly seen in male around 40 years old.The lesions were mostly equal or high density on CT,equal or low signal intensity on T 1WI and closed to grey matter signal intensity on T 2WI.Tumors were enhanced obviously on contrast-enhanced scans,necrosis of center of focus was seen in 1/3 cases.The lesions were single or multiple.Conclusion The accuray diagnosis of intracranial primary malignant lymphoma can be done by combined clinical data with imaging features.

Symptomatology and medical history taking are the focuses of diagnostics study. With the improvement of teaching and learning, students can basically master the standard process of clinical inquiry of chief complaints to family history efficiently. However, due to the lack of systematic learning of clinical diseases, it's difficult for most students to form a targeted, logical and speculative inquiry thinking pattern, so they are unlikely to write a medical history of present illness that reflects their thinking on disease inclusion and exclusion. In this study, we come up with the characteristics and defects of symptomatology teaching at present, and put forward a new idea of symptomatology teaching by introducing clinical manifestations of diseases and examples of inquiry.

Objective To investigate the effect of clinical pathway on inpatients for rehabilitation. Methods The hospital expense, the ex-pense for medicine and length of stay in hospital were compared in patients with cerebral hemorrhage, cerebral infarction, spinal cord injury and spastic cerebral palsy before (January 1st, 2012 to April 30th, 2013) and after (May 1st, 2013 to December 31st, 2015) clinical pathway implementation, 50 inpatients for each disease, totally 200 patients. Results The length of stay in hospital reduced after of clinical pathway implementation, in all the diseases (t>5.226, P<0.001), with the decrease of hospital expense for cerebral infarction (t=3.327, P<0.001). There was no significant increase in any expense in the disease (t<1.777, P>0.05). Conclusion The implementation of rehabilitation clinical pathway can reduce the length of stay in hospital for the patients rehabilitation in hospital, without increasing their cost.

Objective This study was designed to establish and optimize the two-dimensional gel electrophoresis (2-DE) for the proteome analysis of human Transitional Cell Carcinomas of the Urinary Bladder ,and to established the two-dimensional gel electrophoresis (2-DE) profile of human Transitional Cell Carcinomas of the Urinary Bladder(TCC) of different stages in carcinogenic process. The basic aim is to perform differential analysis and provide a basis for identifying carcinogenesis-associated protein of TCC.Methods After obtaining samples of the normal,invasive tissue,noninvasive tissue,a series of methods Such as sample preparation,electrophoresis parameters,gel concentration,PDQuest 2DE analysis software,peptide mass fingerprint (PMF,MALDI-TOF-MS)were used to analyses these tissues.Results The good 2DE patterns of different tissues including resolution and reproducibility were obtained. After coomassie Brilliant Blue R-250 staining, the average matching ratio was 76%.There was a good reproducibility of spot position in 2-DE map, with average deviation in IEF direction of (0.98?0.26) mm, while in SDS-PAGE direction it was(1.23?0.22) mm. The 2-DE patterns with good quality had been obtained. Ten protein spots chosen randomly from different stages were identified by PMF, some of which were involved in the cell proliferation, differentiation, cycle regulation, and tumor occurrence, such as Heat shock protein-27,SFN etc.Conclusions The results provide a fundamental basis for further study of mechanism of human Transitional Cell Carcinomas of the Urinary Bladder and screening its specific marker.

AIM: To establish a method for simultaneously determining chlorogenic acid,puerarin and paeoniflorin in Kangganning Mixture(Flos Lonicerae japonicae,Radix Puerariae Lobatae,Radix Paeoniale rubra,Fructus Forsythiae,etc.). METHODS: A multiple wavelength HPLC method was devoloped.The analysis was performed on an Agilent Zorbax SB C_(18) column(250 mm?4.6 mm,5 ?m) with acetonitrile-0.1%H_3PO_4(11∶89) as the mobile phase.The detection wavelengthes were monitored at 324 nm,254 nm and 230 nm for chlorogenic acid,puerarin and paeoniflorin,respectively.The flow rate was 1.0 mL/min and the column temperature was at 30 ℃.(RESULTS:) The calibration curves of chlorogenic acid,puerarin and paeoniflorin showed good linearity at the ranges of 0.179-2.864 ?g,0.071 55-1.144 8 ?g and 0.372 5-5.96 ?g,respectively.The average recoveries were(100.7%,) 103.3%,102.6% with RSD of 2.2%,2.4%,1.9%,respectively. CONCLUSION: The method is simple,accurate and reproducible for determining chlorogenic acid,puerarin and paeoniflorin in Kangganning Mixture.