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

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 identify the differentially expressed genes in Uigur and Kazak patients with and without type 2 diabetes mellitus.Methods The differentially expressed cDNA bands were isolated by fluorescent mRNA differential display from peripheral blood leucocyte of the Uigur and Kazak patients with type 2 diabetes mellitus and the normal controls. After being cloned, all cDNA fragments were sequenced, then underwent sequence analysis, homogenous comparison,and Northern blot analysis. Results Z5、Z8、Z15 differentially expressed cDNA fragments were found.They were over-expressed in the normal controls and were lower or scarced in the Uigur and Kazak patients with type 2 diabetes mellitus. They were selected for sequencing and hybridization. Z5、Z8 showed highly homologous to cellular repressor of E1A-stimulated genes,Z15 are unknown. Conclusion Three differentially expressed genes may have a potential relation with type 2 diabetes mellitus.

Objective To report two cases of mitochondrial disease presenting with marked gastrointestinal symptoms and mild myopathy without involvement of central nerve system as to investigating the diagnostic rules of the disease Methods Case 1 is a 13 year old boy who had suffered from recurrent vomiting with mild diarrhea and fatigue for 7 years Physical examination showed mild weakness of both legs without ophthalmoplegia and cognitive defects Radiological examination revealed gastroptosis with excessive peristalsis of small intestine Case 2 is of a 6 year old boy who showed repeatedly the recurrent vomiting, intestinal pseudo obstruction and persistent muscle weakness over 1 year and 7 months There were no ophthalmoplegia. Barium clysis showed marked dilatation of colon. Cranial MRI in case 1 and CT in case 2 were unremarkable Muscle biopsies were performed in both cases and specimens were studied using routine histological and histochemical staining as well as using ultrastructural examination mtDNA analysis were performed in both cases Results Histological and enzym histochemical staining showed RRF and SDH positive fibers in both cases. Electron microscopic studies revealed mitochondrial proliferation with paracystalline inclusions in the muscle fibers with mild accumulation of lipid and glycogen. Genetic study revealed point mutation in mtDNA 3243A→G in case 1 and mtDNA3271T→C in case 2 Conclusion Mitochondrial cytopathy might be diagnosed upon the myopathological and genetic findings Different from MNGIE, Leigh disease and MELAS, the main clinical features were gastrointestinal symptoms, mild muscle weakness and no involvement of central nerve system in our patients, thus the mitochondrial entero myopathy, a new clinicopathological type of mitochondrial disorders, might be considered Since the muscle weakness was usually overlooked in patients with diarrhea, the mitochondrial cytopathy occurring should be considered with more attention as to elucidating the causes of children who have chronic gastrointestinal symptoms

Objective To evaluate the prevention and treatment effect of butylphthalide injection in delayed encephalopathy after severe carbon monoxide poisoning(DESCMP).Methods One hundred and fiftyeight cases with severe carbon monoxide poisoning (SCMP) were randomly assigned to 2 groups.Observation group (n =83), butylphthalide treatment when SCMP was diagnosed;Control group (n =75), with no butylphthalide therapy.Incidence rate in 3 month, drug amount and the clinical effects of butylphthalide were observed.Results Observation group occurred 5 DEACMP, with the incidence rate of 6.58％, and control group occurred 14 cases,with the incidence rate of 18.67％ ,significant difference was observed in two groups(x2 =154.204,P＜0.05).There were also significant differences in the glasgow coma scale(14.00±1.89 vs.13.00 ±3.30,t =1.780, P＜ 0.05) and mini-mental state examination (26.20± 2.89 vs.19.57 ± 7.12, t =2.003, P＜0.05) between the two groups.Conclusion Early treatment with butylphthalide can decrease the incidence rate of DEACMP.

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.

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

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℃ and 45℃ 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.

Objective To discuss the values of three screening methods for the detection of early breast cancer,and to analyze the features of the screening cancer.Methods The first screening of breast cancer were performed in 5307 women who aged from 20 to 76 years with median age of 49 years.The three screening methods included physical examination with ultrasound and mammography,physical examination with mammography and mammography only.The rate of recall,biopsy,cancer detection of three methods were analyzed and the mammographic findings were reviewed Chi-square test or Fisher's exact test were used for the statistics.Results The recall rates were 4.90% (49/1001),6.90%(166/2407)and 4.48% (85/1899) in three methods respectively,the biopsy rates were 1.60% (16/1001),1.04% (25/2407) and 0.63%(12/1899),the cancer detection rates were 0.50% (5/1001),0.17% (4/2407) and 0 (0/1899).There were statistical differences among the three groups (X2=12.99,6.264,8.764,P < 0.05).Physical examination with ultrasound and mammnography had the highest cancer detection rate,ten breast cancers were detected and 8 were early stage breast cancer.Of seven cancers detected by mammography,only two were found by ultrasound.A cluster of calcifications were found in 2 cases,linear calcifications in 2 cases.One case presented as a asymmetric density,one as a asymmetric density with calcifications,one as multiple nodules with a duster of calcifications.Two breast cancers presented as asymmetric density were missed on mammography and diagnosed correctly after retrospective review.Conclusion Physical examination with ultrasound and mammography is the best method for breast cancer screening.The breast cancer can be detected by mammography earlier than other methods.