Objective To test the status of Keap-1/Nrf-2 pathway in cisplatin-resistant Skov3/DDP cells,and the contribution of these molecular in cisplatin (cis-pt) resistance.Methods Skov3/DDP was divided into four groups:blank,cis-pt,Keap-1 up-regulated and Keap-1 plus cis-pt.Keap-1 was up-regulated in Skov3/DDP cells by trans-faction with pFlag-CMV-Keap-1 plasmid which was validated by expression of flag and increase of Keap-1 level using Western blot.The effect of Keap-1 up-regulation on Nrf-2 re-distribution was analyzed by Western blot.FCM was employed to detect apoptosis.ROS level was measured by DHE staining.Results The expression of Keap-1 was decreased obviously in Skov3/DDP cells compared with Skov3 cells (P＜0.05),Nrf-2 in Skov3/DDP cells detected mainly in nucleus.Then,the Keap-1 was successfully up-regulated (3.5 fold) by exogenous expression of pFlag-CMV-Keap-1 plasmid,as a result,nucleus portion of Nrf-2 was significantly reduced (0.2 fold).Next,the FCM results showed significant increase of apoptosis in Keap-1 up-regulated Skov3/DDP cells after cis-pt administration (P＜0.05).Finally,ROS level was significantly raised in Skov3/DDP cells treated with cis-pt plus Keap-1 up-regulation.Conclusions Up-regulation of Keap-1 reduced the nucleus portion of Nrf-2 and reverses cisplatin resistance in Skov3/DDP cells.

Objectve To study director-leading teaching method,raise the students' main body consciousness and innovation spirit and improve the teaching effect of microbiology test.Method 158 students majored in laboratory medicine of Grade 2013 were divided in two groups.80 from class 1 were in test group,and the rest from class 2 were served as control group.The clinical microbiological specimens were collected and identified;the experimental class was designed with the experimental teaching mode,while the control group used the traditional experimental teaching mode.Through the experimental evaluation and questionnaire survey,the difference between new teaching model and the traditional teaching mode were evaluated.SPSS 21.0 software was used in the collected data for processing.Result The theoretical results of the experimental class were (88.65 ± 7.64) and the practical skills were (89.72 ± 7.69) and the control group were (79.88 ± 10.64) and (80.55 ± 9.06).The scores of the experimental class were significantly higher than those of the control class,and there and statistically significant difference between the scores of two classes.T values were 6.851 and 5.938 and P values were 0.000 and 0.000 respectively.Survey results show that:90％ of the students agree with the new teaching methods.Conclusion The teaching result of the new teaching mode is superior to the traditional teaching mode.

A mathematical model of electro-chemical cell was induced according to its electric model. Based on the mathematical model, Fourier transform (FT) was applied to separate the charging current from Faradaic current in fast scanning voltammetric signal. The proposed method was investigated with both simulated voltammogram and experimental voltammogram. The results indicate that the proposed methods can successfully separate the charging current from Fradaic current, and the result is consistent with theoretical model.

OBJECTIVE:To prepare chloramphenicol and ephedrine nasal gel and establish its quality control method.METHODS:Gel was prepared with chloramphenicol and ephedrine hydrochloride as principal agents and with carbomer-940 as base.The contents of the principal agents were determined by HPLC and the stability of the gel was investigated as well.RESULTS:The preparation was transparent colloid.The linear ranges of chloramphenicol and ephedrine hydrochloride were 62.5～1 000 and 31.25～500 ?g?L-1(r=0.999 9),respectively,with average recovery rates at 99.64%(RSD=0.5%)and 99.57%(RSD=0.7%),respectively.Storing under room temperature,no obvious change was noted for the property of the sample at 12 months.CONCLUSION:The preparation technique of the gel is simple and feasible,and its quality is stable and controllable.

To study the in situ intestinal absorption of five oligosaccharides contained in Morinda officinalis How. (sucrose, kestose, nystose, 1F-Fructofuranosyinystose and Bajijiasu). The absorption of the five oligosaccharides in small intestine (duodenum, jejunum and ileum) and colon of rats and their contents were investigated by using in situ single-pass perfusion model and HPLC-ELSD. The effects of drug concentration, pH in perfusate and P-glycoprotein inhibitor on the intestinal absorption were investigated to define the intestinal absorption mechanism of the five oligosaccharides in rats. According to the results, all of the five oligosaccharides were absorbed in the whole intestine, and their absorption rates were affected by the pH of the perfusion solution, drug concentration and intestinal segments. Verapamil Hydrochloride could significantly increase the absorptive amount of sucrose and Bajijiasu, suggesting sucrose and Bajijiasu are P-gp's substrate. The five oligosaccharides are absorbed mainly through passive diffusion in the intestinal segments, without saturated absorption. They are absorbed well in all intestines and mainly in duodenum and jejunum.
Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Intestinal Absorption ; Intestine, Small ; metabolism ; Male ; Morinda ; chemistry ; Oligosaccharides ; chemistry ; pharmacokinetics ; Perfusion ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo construct a cDNA library of osteosarcoma SAOS-2 cells and screen the proteins interacting with the bone morphogenetic protein-2 (BMP-2) to explore the role of BMP-2 in the development of prostate cancer.

METHODSThe total RNA was extracted from SAOS-2 cells, from which poly(A)(+)RNA was purified to generate the cDNA using RT-PCR with primer SMARTIII and primer CDSIIIoligo(dT). The library was constructed and screened by cotransformation of the double-stranded cDNA (dscDNA) and linearized pGADT7-Rec with the bait plasmid to the yeast AH109. The proteins interacting with BMP-2 were screened by AH109 mating with the bait strain Y187, and the interaction was confirmed using far-Western blot analysis.

RESULTSThe cDNA library of human osteosarcoma SAOS-2 cells was constructed, which was characterized by high diversity and sufficient capacity. The dscDNA size range was between 250-5000 bp, with a cotransformation efficiency of 4.3 x 10(5) and recombination efficiency of 1.9 x 10(6), and 4.3 x 10(5) clones were screened. The mating efficiency was 32% and 1.0 x 10(6) clones were screened by the bait of BMP-2, and 4 positive clones was obtained and sequenced.

CONCLUSIONThe diversity and capacity of the cDNA library meet the needs for screening genes related to prostate cancer.

Bone Morphogenetic Protein 2 ; genetics ; Cell Line, Tumor ; Gene Library ; Humans ; Male ; Osteosarcoma ; genetics ; Prostatic Neoplasms ; genetics ; RNA, Neoplasm ; genetics ; Two-Hybrid System Techniques

· AIM: To explore the dynamic expression and correlation among telomerase catalytic subunit (TERT), proliferating cell nuclear antigen ( PCNA) and antiapoptosis protein Bd-2 which relate to cell proliferation in epiretinal membrane of rat traumatic proliferative vitreoretinopathy(PVR).· METHODS: S-P technique was applied for immunohistochemical staining of epiretinal membrane of traumatic PVR with TERT, PCNA and Bcl-2 antibody. HE staining was also carried out. The staining results were analyzed with image analysis system.· RESULTS: The positive rate and average A of PCNA protein were upregulated at first and then down-regulated, with the peak value in 14d Group, which was significantly different from those in 7d Group and 28d Group.The positive rate and average A of TERT and Bcl-2 were also upregulated at first and then down-regulated, with the peak value in 14d Group and 21d Group, which were significantly different from those in 7d Group. There was significant correlation among PCNA, Bcl-2 and TERT protein expression (P≤0.01).· CONCLUSTON: TERT and Bcl-2 take part in the regulation of proliferative cells in epiretinal membrane of traumatic proliferative PVR, with high correlation with the dynamic changes of cell proliferation.

Objective To investigate the improvement of postpartum pelvic floor by rehabilitation training assessed with three-dimensional transperineal ultrasound . Methods One hundred cases of healthy postpartum women were randomly divided into two groups :control group and training group .The control group received the customary education ,and the training group received pelvic floor rehabilitation training . At 6 and 12 weeks postpartum ,levator hiatus area ,thickness of the levator ani muscle ,bladder neck mobility ,and bladder posterior horn were measured with three-dimensional transperineal ultrasound in all the subjects . Meanwhile ,the muscle strength situations were tested . Results At 12 weeks postpartum ,the anal levator hiatal area ,bladder neck mobility and bladder posterior horn in the training group were lower than those of the control group[ ( 21 .6 ± 3 .2) cm 2 vs ( 25 .6 ± 2 .4 ) cm 2 ,( 27 .9 ± 5 .3) mm vs ( 31 .5 ± 5 .9) mm ,( 126 .3 ± 21 .2)° vs (135 .3 ± 11 .6)°] ( P < 0 .05) . Compared with control group ,the thickness of the levator ani muscle increased in training group [ ( 13 .6 ± 2 .3) mm vs ( 15 .3 ± 2 .5) mm ] ( P < 0 .05) . The incidence of stress urinary incontinence in the training group ( 5% ) was significantly lower than the control group ( 12 .5% ) at 12 weeks postpartum ( χ2 = 5 .487 , P = 0 .025) . The muscle strength had no significant difference at 6 weeks postpartum . At 12 weeks postpartum ,the pass rate of class Ⅰ muscle fiber was 78 .5% ,and that of class Ⅱ muscle fiber was 83 .3% in the training group ;the pass rate of class Ⅰ muscle fiber was 28 .5% ,and class Ⅱ muscle fiber was 37 .3% in the control group , the improvement was significant at 12 weeks postpartum . Conclusions The result of the transperineal real-time ultrasonographic evaluation of post-natal pelvic floor rehabilitation training has high consistency with the measurement of muscle strength . The ultrasound examination is simple and accurate ,and has highly applicable value in evaluating the effect of post-pelvic rehabilitation training .

ObjectiveTo explore the effect of musicotherapy on agitation behaviors in patients with aged dementia (AD).Methods22 AD patients were treated with different musicotherapies, and examined with Mini-Mental State Examination (MMSE), Cohen-Mansfield Agitation Inventory (CMAI) and self made sociality psychological inventory before and after treatment.ResultsAfter musicotherapy, the incidence rate of agitation behaviors of AD patients decreased significantly ( P<0.001). The incidence rates of agitation behaviors among patients with low, middle and high MMSE scores were significantly different ( P<0.001). After musicotherapy, both depressions of patients' cognitive function and scores of agitation behaviors were significant different compared with that before treatment (P<0.01). Effectual rates of musicotherapy on sociality psychological emotion, communication, language and intercourse of this group patients were 86.36%, 90.91%, 72.73% and 77.27%, respectively.ConclusionMusicotherapy can effectively alleviate the symptoms of agitation behaviors in AD patients.

OBJECTIVETo study the better efficacy of clinic treatment for dermatitis medicamentosa like of trichloroethylene (DMLT) and observe the clinic efficacy of Xuebejing injection for DMLT patients with systemic inflammatory response syndrome (SIRS).

METHODS16 DMLT patients with SIRS were randomly divided into control group (conventional therapy) and xuebijing group (conventional therapy plus xuebijing). We evaluated all the patients with APACHE II before treatment and checked the TNF-alpha in blood at the different time (before treatment and the 7th day of treatment). The total usages, first dosage of medrol and the time of therapy for each group were counted.

RESULTSAll patients were cured, there was no significant difference according to APACHE II and TNF-alpha before treatment in two groups . The level of TNF-alpha of all the patients were decreased markedly, but more significantly in xuebijing group (P < 0.01). Moreover, the treatment group patients were given relatively less total usages, first dosage of medrol and time of therapy (P < 0.01).

CONCLUSIONXuebejing injection combined glucocorticoids can cure DMLT patients with SIRS effectively, and reduce the total usages, first dosage of medrol and time of therapy.

APACHE ; Adolescent ; Adult ; Drug Eruptions ; drug therapy ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Trichloroethylene ; toxicity ; Tumor Necrosis Factor-alpha ; blood ; Young Adult