Objective To investigate the effects of neferine on the proliferation and the P-glycoprotein(P-gp) expression of refractory/relapsed acute leukemic cells and to provide experimental evidence for further clinical use. Methods MTT and immunocytochemistry SABC methods were used respectively to observe the alteration of the proliferation and the expression of P-gp in refractory/relapsed acute leukemic cells after treating with neferine. Results The inhibition ratio on acute leukemic cells of neferine adding adriamycin(ADM) group was significantly higher than that of ADM group (P0.05). Conclusion [WTBZ]Neferine can inhibit the proliferation of refractory/relapsed acute leukemic cells, and reduce the P-gp expression of refractory/relapsed acute leukemic cells and consequently reverse multidrug resistance(MDR).

Objective To study the effects of neferine on the expression of glutathione S-transferase-pi in vitro and to explore the multi-drug resistance reversing mechanism of neferine.Methods 50% inhibition concentration(IC_(50)) of ADM on K562/A02 was determined by MTT method.The transcription of GST-? gene was detected by semi-quantitative RT-PCR and the expression level of GST-? was determined by Western blot after neferine treatment.Results Neferine remarkably enhanced chemosensitivity to ADM of K562/A02 cells.After neferine treatment in day 1,day 3 and day 5,the relative efficiency of K562/A02 to ADM was 9.6%,41.4% and 10.7%,respectively.Semi-quantitative RT-PCR showed that mRNA transcription of GST-? gene was significantly reduced(P

Background and Purpose:Recent studies have shown that survivin is an anti-apoptosis gene,which is involved in carcinogenesis and drug resistance of leukemia.Antisense oligodeoxynucleotide(ASODN) can be used to inhibit the expression of survivin,inducing apoptosis and enhancing the chemosensitivity of leukemic cells.This study was designed to explore the effect of survivin ASODN on the growth,apoptosis,and caspase-3 activity of leukemia cell line K562/A02 with the phenotype of drug resistance.Methods:Survivin ASOND was transfected to K562/A02 cells by liposomal reagent,The rate of inhibition,expression of survivin mRNA,apoptosis,and activity of Caspase-3 were detected by colormetric MTT,RT-PCR,flow cytometry and fluorometer,respectively.Results:Survivin ASODN could inhibite the cell proliferation in a dose and time dependent manner.Compared with controls,expression of survivin mRNA decreased by 36.2%(P

Objective It has been known that NSAIDs can inhibit the growth of tumors through cyclooxygenase, but it is unknown that whether there are other mechanisms involved. It is essential to detect the effect of indomethacin for proliferation and apoptosis in the HCT116 and explore its anti tumor mechanism. Methods Human colon adenocarcinoma cell line HCT116 was treated with different concentration of indomethacin for 24 h and CDK 2,CDK 4, Bcl 2, Bax and p21 WAF1/CIP1 protein were detected by Western blot. Results Indomethacin down regulated the expression of CDK 2, CDK 4, Bcl 2 and up regulated the expression of p21 WAF1/CIP1 , however, the expression of Bax remained unchanged. Conclusions Inhibiting proliferation and inducing apoptosis contribute to the anti tumor activity of indomethacin by down regulating the expression of CDK 2, CDK 4, Bcl 2 and up regulating the expression of p21 WAF1/CIP1 .

Objective To evaluate the efficacy and safety of itraconazole injection in treatment of invasive fungal infection in the patients with multiple organ dysfunction syndrome.Methods Invasive fungal infection in 15 patients with multiple organ dysfunction syndrome was treated with 200 mg itraconazole injection,twice a day on the first two days,then once a day,for 8 to 14 days.During the treatment,the symptoms and signs,the results of chemical detection were recorded.Everyday fungus,sputum,stool smear or culture were performed.Results The recovery rate,effective rate,and fungal clearance in this trial with itraconazole injection were 80%(12/15),86.7%(13/15),and 86.7%(13/15) respectively.The side effect was not found.Conclusion Itraconazole injection is an effective and safe drug against invasive fungal infection in patients with multiple organ dysfunction syndrome.

Multidrug resistance (MDR) plays a major obstacle to successful gastric cancer chemotherapy. The purpose of this study was to investigate the MDR reversal effect and mechanisms of hyperthermia in combination with neferine (Nef) in adriamycin (ADM) resistant human SGC7901/ADM gastric cancer cells. The MDR cells were heated at 42°C and 45°C for 30 min alone or combined with 10 μg/mL Nef. The cytotoxic effect of ADM was evaluated by MTT assay. Cellular plasma membrane lipid fluidity was detected by fluorescence polarization technique. Intracellular accumulation of ADM was monitored with high performance liquid chromatography. Mdr-1 mRNA, P-glycoprotein (P-gp), γH2AX expression and γH2AX foci formation were determined by real-time PCR, Western blot and immunocytochemical staining respectively. It was found that different heating methods induced different cytotoxic effects. Water submerged hyperthermia had the strongest cytotoxicity of ADM and Nef combined with hyperthermia had a synergistic cytotoxicity of ADM in the MDR cells. The water submerged hyperthermia increased the cell membrane fluidity. Both water submerged hyperthermia and Nef increased the intracellular accumulation of ADM. The water submerged hyperthermia and Nef down-regulated the expression of mdr-1 mRNA and P-gp. The water submerged hyperthermia could damage DNA and increase the γH2AX expression of SGC7901/ADM cells. The higher temperature was, the worse effect was. Our results show that combined treatment of hyperthermia with Nef can synergistically reverse MDR in human SGC7901/ADM gastric cancer cells.

A simple rapid detection of antibody to hepatitis delta virus (anti-HDV) in human serum was developed by using double antigen sandwich ELISA. HDV gene fragment encoding HDAg was isolated from a Chinese patient infected with HDV by RT-PCR, and a high-efficient expression HD-PQE31 strain was constructed with the fragment. We obtained high titer and good quality hepatitis delta virus protein purified by Ni-NTA metal-affinity chromatography, which was identified by Western blot and ELISA, then we set up the double antigen sandwich ELISA for detection of anti-HDV in human serum, and the performance of the sandwich ELISA was evaluated in terms of specificity and sensitivity. Results were: 1) The purified HDAg protein's purity was 90%, and its ELISA titer was 1/100 000. 2) 42 anti-HDV positive sera were detected and showed that the sensitivity of sandwich ELISA was higher than that of competitive ELISA (t=2.44, p<0.01). 3) The inhibitory rates for 2 anti-HDV positive sera by the specific HDAg were 74% and 93% respectively. 4) For the assay of specificity, all 60 samples infected by other hepatitis viruses and 30 normal samples were negative for anti-HDV. These results suggested that the double antigen sandwich ELISA with purified recombinant HDAg showed higher specificity and sensitivity, It can be used in routine laboratories to diagnose the HDV infection.

Objective To analyze the dynamic evolution of brain MRI in patients with mitochondrial myopathy,encephalopathy,lactic acidosis,and stroke-like episodes (MELAS) syndrome.Methods A retrospective study was performed on 58 MELAS cases with pathologically and (or) molecularly confirmed diagnosis.MRI were repeated within 60 days after the onset of stroke-like episodes (SLE) and the evolution changes of cerebral lesions were accessed.Brain atrophy index (BAI) was calculated in the remission stage from 31 patients with MELAS,and the correlation between BAI,age and disease duration was analyzed.Results The proportion of lesions expansion,migration and shrink within 30 days after the onset of SLE was 64.1％ (25/39),10.2％ (4/39),17.9％ (7/39),respectively,and 13％ (3/23),21.7％ (5/23),56.5％ (13/23),between 30-60 days after the onset of SLE respectively.In the recovery stage of SLE,the BAI in 31 patients with MELAS was 15.2％ ±2.8％.The correlation coefficient between BAI and the age,total disease course and duration of encephalopathy was 0.329 (P =0.043),0.405 (P =0.012) and 0.649 (P =0.000).Conclusions Brain atrophy in the studied MELAS patients gradually develops and strokelike lesions shrink with progression of the disease.However,the migration of lesions is persistent.

OBJECTIVE: To determine the effect of neferine (Nef) combined with mdr-1shRNA on the expression of mdr/P-gp in K562/A02 cell line. METHODS: MTT assay was used to observe the cell proliferation. The expression level of P-gp was determined by Western blot and the transcription of mdr-1 gene was detected by semi-quantitative RT-PCR. RESULTS: After K562/A02 cells were treated by Nef or mdr-1shRNA alone or both for 24 h, the proliferation of K562/A02 cells was significantly higher in the Nef combined with mdr-1shRNA treatment group than that of Nef or mdr-1shRNA alone group (P<0.01).The expression of mdr-1/P-gp in the Nef with mdr-1 shRNA group was significantly lower than that of Nef or mdr-1shRNA alone group. CONCLUSION Nef enhances the inhibition of mdr-1shRNA expression vector on K562/A02 cell proliferation and on P-gp protein to effectively reverse multidrug resistance induced by mdr-1 gene encoding P-gp.
ATP Binding Cassette Transporter, Subfamily B ; ATP Binding Cassette Transporter, Subfamily B, Member 1 ; genetics ; metabolism ; Benzylisoquinolines ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Humans ; K562 Cells ; RNA, Small Interfering ; genetics ; pharmacology

OBJECTIVE: To investigate the efficacy and safety of oral fludarabine in patients with chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL). METHODS: The patients received oral fludarabine 40 mg/(m2.d) for 5 consecutive days, each treatment lasting 4 weeks. The efficacy was assessed with National Comprehensive Cancer Network (NCCN) criteria for response. RESULTS: Twenty-two patients received the treatment, a median of 4 cycles per patient. The rate of complete response (CR), partial response (PR), and overall response (OR) was 40.9% (9/22), 45.5% (10/22), and 86.4% (19/22), respectively. Among the 17 previously untreated patients, 7 (41.2%) achieved CR and 8 (47.0%) achieved PR. Two of the 5 pre-treated patients achieved CR and the other 2 achieved PR. During a median observation of 24 months, the overall survival rate was 81.8%. The main adverse reactions were myelosuppression and infection. Grade 1 to 3 granulocytopenia was found in 7 (31.8%) patients, and infection in 3 (13.6%) patients. Nonhematologic toxicity was mild. All the adverse reactions were reversible. CONCLUSION The oral fludarabine is effective, safe, and well-tolerated in the patients with CLL/ SLL.
Aged ; Antineoplastic Agents ; adverse effects ; therapeutic use ; Female ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; drug therapy ; Male ; Middle Aged ; Treatment Outcome ; Vidarabine ; adverse effects ; analogs & derivatives ; therapeutic use