The exam under open and close condition is a new mode which is different from traditional close-exam and entire open-exam.It indicates that new exam mode not only conquers limitation of simplex close-exam,but also boosts forwardly study of students and promotes the didactical reform completely.

Targeting induced local lesions in genomes (TILLING) is a reverse genetics method for functional genomics research.It is possible to screen for point mutations in the populations of EMS mutagenesis with highthroughput and lowcost. EcoTILLING a method based on TILLING ,was developed for detecting multiple types of polymorphisms in germplasm collections,such as single nucleotide polymorphism,small deletion and insertion etc.Rice is a very important food crop and a model plant for genome research also. There are complete genome sequence and a lot of other bioinformatics resources about it.So the markerassisted breeding is becoming more and more important in rice breeding. Some issues based on TILLING about identifying germplasm based on gene sequence,EMS mutagenesis breeding,developing functional marker in rice breeding in future were discussed.

Objective To investigate the therapeutic effect and adverse effects of the teniposide-based regimen in patients with primary central nervous system lymphoma (PCNSL). Methods Between March 2011 and July 2013, 16 patients with PCNSL were diagnosed and treated. The clinical characteristics, diagnosis,therapy, results and adverse effects were analyzed. Results Totally 16 patients were enrolled and diagnosed as primary central nervous system diffuse large B-cell lymphoma. All patients received teniposide-based regimen chemotherapy and 9 patients received teniposide plus rituximab. The overall response rate was 87.5 % (14/16), including 10 cases of CR and 4 cases of PR. With a median follow-up of 13.5 months, the progression-free survival (PFS) and overall survival (OS) rates of 2 years were 29.9 % and 66.7 %, respectively. The mainly hematological adverse events were neutropenia, including grade 3 in 4 cases (25 %) and grade 4 just in one case. There was one case of treatment related death. Conclusions The response rate of teniposide-based regimen for PCNSL is promising. The 2 year PFS and OS rates are even higher than results of traditional high-dose methotrexate regimen. The teniposide-based regimen is well tolerated, and the adverse events are acceptable.

Objective To analyze the clinical characteristics , diagnosis, therapy and prognosis of new diagnosed pri-mary gastric diffuse large B cell lymphoma ( PGDLBCL) and to discuss the efficacy of rituximab .Methods Between Jan 2005 and May 2012 , twenty-six new-diagnosed PGDLBCL patients were reviewed retrospectively .The clinical characteris-tics, diagnosis, therapy, results and prognostic factors were analyzed .Results There were 14 males and 12 females.Their age ranged from 25 to 82 (median, 50.1) years old.The most common symptom was stomachache .Treatment strategies were chemotherapy alone ( n=9) [ scheduled as cyclophosphamide , doxorubicin , vincristine and prednisone ( CHOP) and CHOP-like] and chemotherapy combined with rituximab (n=17), followed by radiotherapy of the stomach with or without regional nodes .All clinical and pathological features were similar between the two groups .The median follow-up time was 40 months.The overall response rate was 100%(9/9)in CHOP group, including 55.56%(5/9) CR, and 93.75%(15/16) in RCHOP group including 50%(8/16) CR (P>0.05).The total PFS and OS of 5 years were 60.3%and 74.4%respectively.The PFS in CHOP group and RCHOP group was 66.7% and 58.9%, respectively,and the OS was 66.7%and 84.6%, respectively.Although the OS of RCHOP group was much better than that of CHOP group , there was no sta-tistically significant difference.Univariate analysis showed that IPI (P<0.05) and Lugano staging (P<0.05) were inde-pendent factors of survival in patients with PGDLBCL .Conclusion Chemotherapy could be the first-line therapy of PGDL-BCL.The overall survival rate might be increased by adding rituximab to chemotherapy .The Lugano stage and IPI are im-portant prognostic factors .

Objective To investigate the expression of nestin, a type Ⅵ intermediate filament protein in the glomeruli with foot process effacement and the potential relationship between nestin expression in the kidney and the degree of proteinuria. Method Immunohistochemistry was used to determine the localization of nestin in the kidney samples obtained from needle biopsies of normal human and patients with minimal change disease (MCD). Puromycin aminonucleoside (PAN) nephrosis rat models were established by a single intraperitoneal injection of PAN. Both real time quatitative reverse PCR and Western blot methods were applied to evaluate the levels of nestin expression at day 1, 4, 10 and 20 after PAN injection. Results Immunohistochemistry showed that the expression of nestin in glomeruli of MCD patients was significantly reduced compared with normal samples (0.93±0.08 vs 1.65±0.12, P<0.05) . The mRNA and protein expressions of nestin in the rat kidney were transitorily increased by 1.23 folds and 1.48 folds of control group (NC) after 1 day of PAN injection (P<0.05), then decreased quickly in the following days. The mRNA levels of nestin in the kidney were 35.8% and 12.1% of NC after 4 days and 10 days of PAN injection, respectively, (P<0.01) as determined by real time PCR. After 20 days of PAN injury, nestin mRNA expression partly recovered to 65.8% of NC (P< 0.05 ). The protein levels of nestin detected by Western blot presented the similar trend, which were 77.0%, 58.0% and 83.4% of NC after 4 days, 10 days and 20 days of PAN injection, respectively (P<0.05). The degree of proteinuria in puromycin aminonucleoside nephrosis rats was negatively correlated with both mRNA and protein levels of nestin in the kidney(r=-0.667,P<0.05 and r=-0.621 ,P<0.05, respectively). Conclusions The expression of intermediate filament protein nestin is down-regnlated in the kidney characterized with foot process effacement and negatively correlated with the degree of proteinuria in puromycin aminonucleoside nephrosis rats. Nestin may play a potential role in modulating the structure and function of podocyte.

Objective To discuss the efficacy of application combination Of minimal immunosuppressive drugs in chronic allograft nephropathy after renal transplantation. Methods Data were drawn from the First Hospital of Tsinghua University.From September 1,2004 to July 1,2006,31 cadaver kidney transplantations were performed using triple immunosuppression with tacrolimus(n=31)and MMF plus steroids before using new strategy.The new strategy is Rapamycin+tacrolimus+MMF+Prednisone.The serum ereatinine,GFR(ml/min/1.73 m2)and 24-hours urine protein before and after 12 months of using lOW dose combination of calcineurin inhibitors,MMF,Rapamune,Predsone and Q80 were recorded.During this time,the concentration of tacrolimus,rapamune were monitored as well. Results After 12 months follow-up,the serum creatinine of 28 patients were decreased from(300±21)μmol/L to(215±38)μmol/L.GFR(ml/min/1.73m2)was elevated from 42.54±2.95 to 49.98±3.05.Three patients whose serum creatinine was 416-464μmol/L had to take hemodialysis.The 24-hours urine protein(g)of 31 patients below 0.8 g did not increase urine protein during follow-up.One patient's 24-hours urine protein(g)increased from 0.95 to 1.29.The patient and graft survival rate was 100%(31/31),90.3%(28/31)respectively.The rapamune main side effect was hyperlipidemia. Conclusions Rapamune and low dose Tacrolimus+Myeophenolate Mofetil+Corticosteroid could be a safe treatment.It may improve renal function in chronic allograft nephropathy.

Background The construction of tissue-engineered corneal endothelium needs the functional seeding cells,so how to culture a large amount of functional corneal endothelial cells (CECs) is an urgent problem to be solved.Objective The aim of this study was to evaluate the role of aqueous humor on bovine CECs in vitro.Methods Aqueous humor of 1.2 ml was collected from the anterior chamber of bovine and sterilized,and the liquid supernatant was obtained.The bovine CECs were isolated from bovine cornea and then cultured in low glucose Dulbecco Modified Eagle Medium with 10％ fetal bovine serum (FBS) in vitro.Aqueous humor was added into the medium with the final concentration of 2.5％,5.0％,l0.0％,15.0％ and 20.0％,respectively,and no aqueous humor was added in the control group.Cell counting kit-8 (CCK-8) assay was used to detect the absorbency value of CECs for the evaluation of cell proliferation.Progression of the cell cycle was analyzed by flow cytometry (FCM).After confluence of the cells was reached,1 ml plastic spear tip was used to scratch the cell single layer,and the cells were incubated consequently in medium with 10％ FBS and with or without aqueous humor for 24 hours.Healing area of the cell single layer was measured.The cells were incubated at a density of 6 × 105 cells/ml and cultured using medium with or without 10.0％ aqueous human for 5 days,and the number of the cells was analyzed by DAPI fluorescence technique.Results Under the phase-contrast microscopy,the confluent CECs showed a slabstone-like and hexagonal appearance.CCK-8 assay revealed that the absorbance values of CECs was significantly different among the various culture groups (F=4.051,P =0.007),and the absorbance value in different concentrations of aqueous human culture groups was significantly higher than that in the control group (P ＜ 0.01).FCM showed that the percentage of the cells in S-G2 phases was (34.80-±3.13)％ in the 10.0％ aqueous humors group and (23.06±1.13)％ in the control group,showing a significant difference (t =-5.729,P=0.005).The scratch test showed that the healing area of the cell signal layer was (0.116±0.019) mm2 in the 10.0％ aqueous humors group and (0.358 ±0.049) mm2 in the control group,showing a significant difference (t =13.842,P =0.000).The density of cells in the 10.0％ aqueous humor group was (1439± 1 10)/field,which was more than (1162±45)/field in the control group (t =-11.020,P=0.000).Conclusions Aqueous humor at the concentration of 10.0％ promote the growth and proliferation of bovine CECs.The result suggests that 10.0％ aqueous humor can be used as a promoting agent during the culture of CECs.

Objective To synthesize trifluoromethyl-substituted mono-carbonyl curcumin analogs and investigate their effects on the proliferation of human lung cancer cells A549 and NCI-H460. Methods Six mono-carbonyl curcumin analogs 1a-1f were synthesized via Aldol condensation using commercially available 2-trifluoromethyl benzadehyde and different ketones (actone, cyclopentanone, cyclohexanone, piperid-4-one, and 1-methylpiperid-4-one). MTT method was used to test the effect of 1a-1f on the proliferation of A549 and NCI-H460 cells. Results Compared with curcumin, most of the mono-carbonyl curcumin analogs 1a-1f exhibited anti-proliferation activity on the A549 cells. The values of IC50 were lower than that of curcumin (P＜0.01), and 1a and 1f showed much better activities both on the A549 and NCI-H460 cells, with their IC50 values were much lower than that of curcumin (P＜0.01).Among them, 1f showed better medicinal chemical properties, with the relative molecular mass less than 500, cLogP 5.25, and the polar surface area (PSA) 37.38, which was more better accorded with the Lipinski′s five rules. Conclusion Six mono-carbonyl curcumin analogs 1a-1f were synthesized, and 1f showed much better anti-proliferation activity on A549 and NCI-H460 cells.

Objective To analyze the clinical characteristics and recent curative effect of mantle cell lymphoma (MCL) after conventional treatment.Methods Clinical data of 15 MCL patients admitted in the Affiliated Hospital of Academy of Military Medical Sciences between August 2004 and October 2013 were retrospectively analyzed.Results The median age of those patients was 59 and the male to female ratio was 1.5∶1.Fourteen(93%)cases were in Ann-Arbor stages Ⅲ -Ⅳ, 15 cases (100%)primarily with lymph node involvement,7 cases (47%)with bone marrow involvement,4 cases (27%)with gastrointestinal involvement,and 3 cases (20%)with orbit involvement.Less than 40% expression of Ki-67 was observed in 9 cases (60%),while 6 cases were with more than 40% (40%).One case was blastic variant.First-line therapy was CHOP-like regimens,which were combined with rituximab in 8 of the 15 cases.In this study,the median survival time was 12 months (3 -64),and the overal response rate was 80%after induction chemotherapy.The current survival of 7 /9 cases with less than 40% expression of Ki-67 was 8 -64 months,2 /6 cases with more than 40% expression of Ki-67 was 8 and 9 months,respectively.Conclusion MCL mostly occurs in older males.Extranodal invasion is common in MCL as an aggressive tumor.The efficacy of traditional chemotherapy is currently limited.Blastic variant or high expression of Ki-67 is an adverse prognostic indicator.

The 1H-NMR fingerprints of three different species tibetan medicine sea buckthorn were established by 1H-HMR metabolomics to find out different motablism which could provide a new method for the quality evaluation of sea buckthorn. The obtained free induction decay (FID) signal will be imported into MestReNova software and into divide segments. The data will be normalized and processed by principal component analysis and.partial least squares discriminant analysis to perform pattern recognition. The results showed that 25 metabolites belonging to different chemical types were detected from sea buckthorn,including flavonoids, triterpenoids, amino acids, carbohydrates, fatty acids, etc. PCA and PLS-DA analysis showed three different varietiest of sea buckthorn that can be clearly separated by the content of L-quebrachitol, malic acid and some unidentified sugars, which can be used as the differences metabolites of three species of sea buckthorn. 1H-NMR-based metabonomies method had a holistic characteristic with sample preparation and handling. The results of this study can offer an important reference for the species identification and quality control of sea buckthorn.
Hippophae ; metabolism ; Magnetic Resonance Spectroscopy ; methods ; Medicine, Tibetan Traditional ; Metabolomics