Objective To investigate the reversal effect of the monomer of traditional Chinese medicine on muhidrug resistance(MDR) and its possible mechanism in K562/ADM cell line in vitro. Methods With different concentrations of baicalin, geniposide administered to K562/ADM cells, the proliferation of K562/ ADM cells was detected by the MTY assay. Expression of mdr-1 mRNA, Topo Ⅱ mRNA was measured by semi-quantitive RT-PCR. Results Thatbaicalin and geniposide could increase the sensitivity of K562/ADM cells to adriamycin, multiples of reversion were 1.95 times and 1.46 times. The proliferation of K562/ADM cells was in-hibited obviously by baicalin and geniposide, the level of mdr-1 mRNA expression was down-regulated and the Topo Ⅱ mRNA was up-regulated(P<0.01 ). Conclusion Baicalin and geniposide may reverse the multi-drag-resistance of K562/ADM cells, which was related to the down-regulation of mdr-1 expression and up-reg-ulation of Topo Ⅱ beta expression.

Objective To explore molecular mechanism of expression of vascular endothelial growth factor (VEGF) mRNA and secretion of VEGF protein in HL-60 cells induced by all-trans refinoic acid (ATRA). Methods MTr method was used to detect the proliferation of HL-60 cells induced by ATRA,cell cycle and CD11b expression in HL-60 cells were detected by flow cytometry. Expression of VEGF, c-myc, by-poxia-inducible factor(HIF)-lα, matrix metalloproteinase (MMP)-9 and MMP-2 mRNA were detected by semi-quantitative RT-PCR. VEGF protein in HL-60 cells culture supernatant was measured by ELISA before and after being induced by ATRA. Results After treatment with ATRA,the proliferation of HL-60 cells was obviously inhibited, CD11b expression increased, trend of granulocyte directional differentiation emerged, and differentiation degree was increasd(P <0. 05) ;expression level of c-my and VEGF mRNA was down-regulated (P < 0. 05), but expression level of HIF-1α mRNA was up-regulated (P < 0. 05). VEGF protein level in HL-60 cells culture supernatant was decreased by blocking the expression of MMP-9 or MMP-2(P <0. 05).Conclusion VEGF expression has positive correlation with c-myc expression,but has negative correlation with HIF-1α expression. MMP-9 and MMP-2 may be the main factors regulating VEGF secretion in HL-60 cells.

Objective To study the molecular mechanism of different sensitivities to apoptosis induced by low concentration of As2O3 in PML-RARα negative HL-60 cells and PML-RARα positive NB4 cells. Meth-ods NB4 and HL-60 cells were cultured with As2O3 for 1 to 4 days; cell proliferation were detected by MTT method; the apoptosis was detected by flow cytometry,Bcl-2,Bax and Fas mRNA were determined by RT-PCR. Results The proliferation of NB4 cells was inhibited obviously by As2O3(1.0 μmol/L)with the induction of apoptosis( P <0.05) ,which was accompanied by the down-regulation of Bcl-2 mRNA expression( P <0.05)and the ratio of Bcl-2/Bax(P <0.05), but there was no obvious variation of Bax and Fas expression( P >0.05). Inhibition of proliferation and apoptosis were not obvious in PML-RARα negative HL-60 cells induced by low concentration As2O3 ( P >0.05), and there was no obvious variation of Bcl-2, Bax, Fas mRNA expres-sion or Bcl/Bax ratio( P >0.05). Conclusion The ratio of Bcl-2/Bax is contributed to the different sensitiv-ities of PML-RARα negative HL-60 cells and positive NB4 cells induced by low concentration of As2O3.

OBJECTIVETo evaluate the preliminary curative effect of interspinous injections for the diagnosis and treatment of back pain caused by lumbar kissing spine (Baastrup's disease) under fluoroscopically guiding.

METHODSFrom November 2011 to March 2013,17 patients with back pain caused by Baastrup's disease were treated with fluoroscopically-guided interspinous injections, including 7 males and 10 females with an average age of 49.6 years old ranging from 40 to 71 years old; the duration of the disease ranged from 2 to 5 years with a mean of 3.7 years. The visual analogue scale (VAS) and the lumbar segments range of motion (ROM) was analyzed at pre-operation, 2 days, 3 months and final followed-up after operation, the effects were evaluated with modified Macnab standard.

RESULTSAll patients were follow-up from 6 to 10 months with an average of 7.6 months. The pre-operative VAS was 6.41 +/- 0.94, the postoperative VAS at different time points improved significantly comparing with pre-operation,and the differences were statistically significant (P < 0.01). There was no significant difference in VAS at different time points after operation (P > 0.05). The ROM of operated segment and adjacent segment was (4.88 +/- 0.86) degrees and (6.82 +/- 0.73) degrees respectively at pre-operation. The postoperative operated segment ROM at different time points improved significantly comparing with pre-operation, and the differences were statistically significant (P < 0.05). Compared with pre-operation, there was no significant difference in adjacent segment ROM at different time points after operation (P > 0.05). According to modified Macnab, the result was excellent in 6 cases, good in 7 cases, fair in 3 cases and poor in 1 case.

CONCLUSIONFluoroscopically-guided interspinous injections is an effective method for the diagnosis and treatment of Baastrup's disease. The method has advantages of simple operation, minimal invasion and safety, satisfactory short-term and medium-term therapeutic effect; it can also effectively lessen the pain of lumbar and back.

Adult ; Aged ; Anesthetics, Local ; administration & dosage ; Female ; Humans ; Low Back Pain ; diagnosis ; diagnostic imaging ; drug therapy ; Lumbar Vertebrae ; diagnostic imaging ; drug effects ; Male ; Middle Aged ; Nerve Block ; Spinal Diseases ; diagnosis ; diagnostic imaging ; drug therapy ; Tomography, X-Ray Computed

Background: In China, most patients with nasopharyngeal carcinoma (NPC) are diagnosed at a late stage and con-sequently have a poor prognosis. This study aimed to investigate potential factors associated with the clinical stage of NPC at diagnosis. Methods: Data were obtained from 118 patients with early-stage NPC and 274 with late-stage NPC who were treated at Sun Yat-sen University Cancer Center between August 2014 and July 2015. Patients were individually matched by age, sex, and residence, and a conditional logistic regression model was applied to assess the associa-tions of clinical stage at diagnosis with socioeconomic status indicators, knowledge of NPC, physical examinations, patient interval, and risk factors for NPC. Results: Although knowledge of early NPC symptoms, smoking cessation, and patient interval were important fac-tors, the number of cigarettes smoked per day, motorbike ownership, and physical examination exhibited the strong-est associations with the clinical stage of NPC at diagnosis. Compared with smoking fewer than ten cigarettes a day, smoking 10–30 cigarettes [odds ratio (OR) 4.03; 95% confidence interval (CI) 1.11–14.68] or more than 30 cigarettes (OR 11.46; 95% CI 1.26–103.91) was associated with an increased risk of late diagnosis. Compared with not owning a motorbike, owning a motorbike (OR 0.38; 95% CI 0.23–0.64) was associated with early diagnosis. Subjects who under-went physical examinations were less likely to receive a late diagnosis than those who did not undergo examinations (OR 0.50; 95% CI 0.28–0.89). However, indicators of wealth were not significant factors. Conclusions: Initiatives to improve NPC patient prognosis should aim to promote knowledge about early symptoms and detection, health awareness, and accessibility to health facilities among all patients, regardless of socioeconomic status.

OBJECTIVETo investigate the effects and regulatory mechanism of ILK on angiogenesis in hypertrophic scar.

METHODSThe human scar microvascular endothelial cells (HSMECs) were isolated from 6 patients' hypertrophic scar in vitro. The HSMECs with good condition in 2nd to 4th generation were selected as experimental objectives. (1) HSMECs were divided into the blank control group (treated with routine culture), negative control group (treated with only Lipofectamine 2000), LY294002 group (incubated with 50 nmol/L LY294002), ILK siRNA group (incubated with 20 nmol/L ILK siRNA). RT-PCR and Western Blot were used to detect the expression of ILK mRNA and its protein after transfecion for 48 h. (2) The digested HSMECs of four groups were resuspended with DMEM without serum and then seeded onto the upper compartment of transwell insert which contained complete medium in its lower compartment. The cell migration experiment was stopped in 10 h and then the migrated cells were counted to analyze the effects of different interventions on the migration ability of HSMECs. (3) The thawed ECMatrix was put into each well of pre-colled 48-well tissue culture plate, and then the plate was put into the incubator at 37 degrees C to make it to become gel. The HSMECs of four groups were seeded onto the surface of the ECMatrix gel and were put into incubator. Eight random view-fields per well should be valued by the sheet of pattern recognition about angiogenesis after 8 hours to evaluate the ability of angiogenesis in vitro between four groups.

RESULTS(1) The expression of ILK mRNA (ILK mRNA = 0.829 +/- 0.109, t = 13.151, P = 0.006) and protein (ILK protein = 0.096 +/- 0.049, t = 36.656, P = 0.000) were both inhibited obviously in ILK siRNA group compared with the blank control group (ILK mRNA = 0.829 +/- 0.109, ILK protein = 1). And, the expression of ILK in LY294002 group was slightly lower than that of black control group, but there was no statistical difference. (2) The number of migrated cells in ILK siRNA group (88.111 +/- 3.079) and LY294002 group (138. 667 +/- 2.404) were respectively lower than that in blank control group (322.333 +/- 3.712, P < 0. 05) in 10th hour. (3) Compared to blank control group (4.333 +/- 0.191), the ability of angiogenesis in vitro decreased significantly ILK siRNA group (2.625 +/- 0.125) and LY294002 group (3.125 +/- 0.250), in which, the vascular network structures were not formed perfectly in 8th hour (P < 0.05).

CONCLUSIONSThe ability of HSMECs' migration and angiogenesis in vitro are inhibited significantly when the expression of ILK is down-regulated. It reveals that ILK may play an role in the regulation of scar angiogenesis.

Cell Movement ; Cell Proliferation ; Chromones ; pharmacology ; Cicatrix, Hypertrophic ; enzymology ; pathology ; Endothelial Cells ; cytology ; drug effects ; Humans ; Lipids ; pharmacology ; Morpholines ; pharmacology ; Neovascularization, Pathologic ; etiology ; pathology ; Protein-Serine-Threonine Kinases ; genetics ; physiology ; RNA, Messenger ; analysis ; RNA, Small Interfering ; metabolism

This research aimed at studying the effects of irrigation and rhizome length on the survival of ratio, yield and quality of Glycyrrhiza uralensis in wild tending condition. Employed the split-block design to carry out the field experiment, sampled with the quadrat method to measured the relative growth indexes and to estimate the yield, used the HPLC (high performance liquid chromatog- raphy ) method to measure the glycyrrhizin in the rhizome and adventitious root of the G. uralensis in this study. The quantity of the adventitious roots and the survival ratio were increased significantly as the length of the rhizome increased (P < 0.01), but the length of the rhizome had no remarkable effect on the content of glycyrrhizin. The average content of the glycyrrhizin in the adventitious root and rhizome could reach 3.03% and 2.12% after 3-year wild tending, respectively, and this results indicated that the quality of the glycyrrhiza using this method was much better than that from cultured glycyrrhiza with the reproducing method of seeding. so using the rhizome as reproductive material to produce the glycyrrhiza under the wild tending condition could get the high quality glycyrrhiza quick- ly and steadily, this phenomenon could be explained by the Hypothesis of synthetic inertia of the medicinal components from the wild material of G. uralensis. But the maximum yield with this method was just more than 945 kg x hm(-2) in this study. So the further work of how to increase the yield in the practical application with the method found in this study need to be done in the next research.
Agricultural Irrigation ; Culture Techniques ; methods ; Glycyrrhiza uralensis ; growth & development ; metabolism ; Glycyrrhizic Acid ; metabolism ; Rhizome ; growth & development ; Survival Analysis

OBJECTIVETo establish a stable H460 cell line with Nrf2 down-regulation to study the role of Nrf2 in Oxaliplatin resistance.

METHODSNrf2 down-regulated H460 cell line was obtained by transfecting cells with pRS-hNrf2 and followed by screening for Nrf2 expression by Western blotting. The cell lines were further characterized by analysing cellular GSH levels and cytotoxicity to anti-cancer drugs with MTS.

RESULTNrf2 down-regulated H460 cell lines were established successfully. Cellular GSH level reduced significantly in the H460-C9 and H460-C13 compared to control cells (P(C9)= 0.00249, P(C13)= 0.03944). Down-regulation of Nrf2 in H460 sensitized the anti-cancer effect of Oxaliplatin and Doxorubicin.

CONCLUSIONNrf2 plays an important role in drug resistance. Down regulation of Nrf2 in H460 cell enhances cytotoxicity of Oxaliplatin.

Antineoplastic Agents ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Drug Resistance, Neoplasm ; genetics ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Organoplatinum Compounds ; pharmacology ; Transfection

OBJECTIVETo investigate the changes of biological characteristics of breast cancer cell line by cyclin E expression.

METHODSHuman breast cancer cell line MCF-7 was transfected with cyclin E siRNA vector pEGFP/CCNE2. siRNA-induced silencing of cyclin E was determined by RT-PCR at RNA level and Western blot at protein level. The proliferation of MCF-7 cells and their sensitivity to chemotherapy was measured by CCK-8 assay. The cells were examined by FCM. The cell line was injected into nude mice and the tumor size was measured.

RESULTSThe expression of cyclin E was inhibited in the MCF-7 cells. The relative expression level of cyclin E mRNA was 0.23 +/- 0.05, and that of cyclin E protein was 0.24 +/- 0.05. The cell growth was inhibited by 68.56% +/- 0.08%, and their sensitivity to chemotherapy was increased. Most cells were blocked at G(1) (77.38%), their tumorigenic ability in nude mice was reduced, and the size of tumor formed in mice of the experimental group was decreased than that of controls.

CONCLUSIONInhibition of cyclin E expression in breast cancer cells can block their cell cycle at G(1) phase, reduce their cell growth, differentiation and proliferation, and increase their sensitivity to chemotherapy.

Animals ; Antibiotics, Antineoplastic ; pharmacology ; Antimetabolites, Antineoplastic ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin E ; genetics ; metabolism ; Doxorubicin ; pharmacology ; Female ; Fluorouracil ; pharmacology ; Genetic Vectors ; Humans ; Male ; Mice ; Mice, Inbred ICR ; Mice, Nude ; Neoplasm Transplantation ; Paclitaxel ; pharmacology ; Plasmids ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Tumor Burden