Humans ; Adolescent ; DNA/genetics* ; Genotype

The present study is to elucidate the mechanisms underlying Gleevec-induced apoptosis of chronic myeloid leukemia (CML) K562 cells in vitro. The apoptotic cell death and cell cycle distribution after Gleevec treatment and the effect of PDCD4 siRNA on Gleevec-induced apoptosis of K562 cells were analyzed by flow cytometry. The effect of Gleevec on p-Crkl, caspase-3, PARP and PDCD4 protein levels, and the knockdown efficacy of PDCD4 siRNA were detected by Western blotting. The results showed that Gleevec dramatically suppressed the phosphorylation level of Crkl in a dose-dependent manner and induced significant apoptosis and G0/G1 cell cycle arrest of K562 cells in time- and dose-dependent manners. In addition, Gleevec activated caspase-3 and its downstream substrates PARP, and the caspase pan inhibitor Z-VAD-FMK (50 micromol x L(-1)) markedly reduced Gleevec-induced apoptosis from 47.97% +/- 10.56% to 31.05% +/- 9.206% (P < 0.05). Moreover, Gleevec significantly increased the protein expression of programmed cell death 4 (PDCD4). PDCD4 knockdown by siRNA reduced Gleevec-induced apoptosis from 46.97% +/- 14.32% to 42.8% +/- 11.43%. In summary, Gleevec induced apoptosis in K562 cells via caspase-3 activation.
Amino Acid Chloromethyl Ketones ; Apoptosis ; drug effects ; Benzamides ; pharmacology ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Humans ; Imatinib Mesylate ; K562 Cells ; Phosphorylation ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology

Objective To establish an animal model of dermatophytosis and to evaluate antifungal efficacy on dermatophytosis with this model. Methods Animal models of dermatophytosis were established by inoculating dermatophyte suspension onto abraded skin on the back of guinea pigs. Thirty- eight healthy guinea pigs were randomly and equally divided into 2 groups, namely, Trichophyton mentagrophytes group (infected with T. mentagrophytes), and Microsporum canis group (infected with M. canis), and each group was classified into three subgroups, i.e., itraconazole group treated with oral itraconazole of 4 mg per kilogram body weight per day from day 0 to day 14 after infection, terbinafine group treated with oral terbinafine of 5 mg per kilogram body weight per day from day 0 to day 14 after infection, and untreated group receiving no therapy. The therapeutic effect was evaluated according to skin lesion score and fungal examination results on day 8, 11 and 14 after infection. Results Obvious lesions were observed and fungal examination was positive in untreated, infected pigs on day 8 after infection. In T. mentagrophytes-infecyted pigs, the skin lesion score on day 8, 11, 14 was 9, 1 and 0 in itraconazole group, 8, 5, and 1 in terbinafine group, 48, 52, 40 in untreated group, respectively, and there was significant difference between treated and untreated groups on the three time points (all P<0.01); the mycological cure rates on the above time points were 66.7%, 83.3%, 83.3%, in itraconazole-treated pigs, 83.3%, 83.3%, 83.3%, in terbinafine-treated pigs, 0, 0, 0 in untreated pigs, respectively, with no significant difference between itraconazole and terbinafine group (all P>0.05) but statistical difference between untreated and treated groups (all P<0.01) on all time points. Meanwhile, in M. canis-infected pigs, the skin lesion score on day 8, 11, 14 reached 3, 0, 0 in itraconazole group, 9, 2, 0 in terbinafine group, 46, 47, 39 in untreated group, respectively, and mycological cure rates 83.3%, 83.3%, 83.3% in itraconazole group, 83.3%, 83.3%, 83.3% in terbinafine group, 0, 0, 0 in untreated group, respectively; significant difference was noticed in the two parameters between the treated and untreated groups (all P<0.01) but not between the two treated groups (all P>0.05). Conclusion Itraconazol and terbinafine exhibit similar excellent antifungal activity in routine model of T. mentagrophytes-and M. canis-dermatophytosis.

Objective To report our experience of retroperitoneal laparoscopic extravascular stent placement for nutcracker syndrome.Methods The clinical data of 12 nutcracker syndrome patients (10 males and 2 females;mean age 26 years) who underwent retroperitoneal laparoscopic extravascular stent placement from March 2014 to Febuary 2016 were retrospectively reviewed.The main symptoms were gross hematuria in 8 patients(one with proteinuria)and flank pain was noted in 1 patient.Three male patients had left-sided secondary varicoceles.Ultrasonography and computed tomography showed the left renal vein clamped by the superior mesenteric artery and the aorta.The anteroposterior diameter of the left renal vein in the renal hilum was three-fold than the aortomesenteric area,and the peak velocity ratio of the aortomesenteric area was much faster than the renal hilum.Twelve patients underwent laparoscopic extravascular stent placement under general anesthesia.The preaortic fibrous tissue between the aorta and the superior mesenteric artery was released intraoperatively.Renal vein became fiat when the superior mesenteric artery was elevated.The 6-8 cm extravascular stent was set on the surface of the renal vein to prevent the compression.Results Stenting was successfully accomplished in all 12 patients.Mean operative time was 62 min (50-125 min),estimated blood loss was 35 ml(20-100 ml),and the hospital stay after operation was 8 days (6-12 days).Three patients had a transient orthostatic intolerance,and they were cured by conservative treatment.With a mean follow up of 14 months (5-30 months),symptoms of hematuria and flank pain resolved in 7/8 and 1/1,respectively.Varicoceles were cured in all three patients.One case got partial relief because of recurrent hematuria due to excessive exercise.Ultrasonography showed that extravascular stent was in the right place,and the angle between abdominal aorta and superior mesenteric artery became normal.The inner diameter of left renal vein was decreased,and the narrow segment was diminished in diameter meanwhile the blood outflow was smooth.Conclusions Retroperitoneal laparoscopic extravascular stent placement in the renal vein is a safe and effective approach for nutcracker syndrome.

OBJECTIVETo evaluate in vitro effect of abnormal savda munziq (ASMq) on the proliferation and apoptosis of human hypertrophic scar fibroblasts (HSFs).

METHODSHSFs were divided into six groups to receive different treatments as group A (blank control group), group B-E (ASMq in different concentration), and group F(5-Fu). Each group contains six specimens. The HSFs were cultured in vitro. After culture for 48 hours, the CCK8 test and flow cytometry methods were used to detect the proliferation, cell cycle and apoptosis.

RESULTSThe proliferation of HSFs in the B, C, D and E groups was inhibited at G2/M period, while it was inhibited at G0/S period in group F (P < 0.05). The inhibition effect of ASMq (0.1-1.0 mg/ml) on the fibroblasts enhanced in a concentration-dependent manner. Flow cytometry analysis with annexin V-FITC and PI staining confirmed the apoptotic. When HSFs were exposed to ASMq at 1.0 mg/ml (group E) for 48 h, the percentage of apoptotic cells increased to (43.7 +/- 2.58)%, which was significantly higher than that of blank control group (2.2 +/- 0.59)%. The induced apoptosis effect was also increased in a concentration-dependent manner.

CONCLUSIONASMq has a inhibitory effect on the proliferation and an enhancement effect on the apoptosis of fibroblast. ASMq could be used as an effective drug for treatment of hypertrophic scar.

Apoptosis ; Cell Cycle ; drug effects ; physiology ; Cell Division ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; pathology ; Fibroblasts ; cytology ; drug effects ; Flow Cytometry ; Humans ; In Vitro Techniques ; Medicine, East Asian Traditional

Objective To make up an exercise prescription based on traditional Chinese medical training (EP-TCMT) for patients with stable chronic obstructive pulmonary disease (COPD).Methods Eighty-five pa- tients with stable COPD were randomly divided into a control group (CG group),a traditional Chinese medicine group ( TC group) and an exercise prescription group ( EP group).The patients in the TC and EP groups were giv- en intensive training for 8 weeks.Their 6 rain walk distance (6MWD) and Borg scale scores were assessed before and after the treatment.Results The 6MWD in the TC group increased from 337.68?59.18 m to 386.14?76.71 m,while those in the EP group improved from 348.00?55.94 m to 425.17?53.22 m.The Borg scale scores in the TC group decreased from 3.14?1.94 to 2.32?1.25,while those in the EP group declined from 3.45?1.84 to 1.72?0.70.Conclusion Making up EP-TCMTs is feasible.Additional treatment was found to improve exercise tolerance and decrease dyspnea in COPD patients.Exercise therapy based on traditional Chinese methods is easy and safe.

AIM: To establish a method for simultaneously determining chlorogenic acid,puerarin and paeoniflorin in Kangganning Mixture(Flos Lonicerae japonicae,Radix Puerariae Lobatae,Radix Paeoniale rubra,Fructus Forsythiae,etc.). METHODS: A multiple wavelength HPLC method was devoloped.The analysis was performed on an Agilent Zorbax SB C_(18) column(250 mm?4.6 mm,5 ?m) with acetonitrile-0.1%H_3PO_4(11∶89) as the mobile phase.The detection wavelengthes were monitored at 324 nm,254 nm and 230 nm for chlorogenic acid,puerarin and paeoniflorin,respectively.The flow rate was 1.0 mL/min and the column temperature was at 30 ℃.(RESULTS:) The calibration curves of chlorogenic acid,puerarin and paeoniflorin showed good linearity at the ranges of 0.179-2.864 ?g,0.071 55-1.144 8 ?g and 0.372 5-5.96 ?g,respectively.The average recoveries were(100.7%,) 103.3%,102.6% with RSD of 2.2%,2.4%,1.9%,respectively. CONCLUSION: The method is simple,accurate and reproducible for determining chlorogenic acid,puerarin and paeoniflorin in Kangganning Mixture.

AIM: To investigate the time - effect relationship between the expression of rhodopsin and recoverin and photoreceptor damage induced by N - nethl - N -nitrosourea ( MNU) .
METHODS: Thirty-six 7-week old Sprague-Dawley ( SD ) rats were intraperitoneally injected with MNU ( 60mg/kg ) and were put to death by dislocation of cervical vertebra 6, 12, 24h; 3, 7d after injection ( 6 per group) , respectively. As a control, six rats were injected with phosphate buffer saline (PBS) 5mL/kg and sacrificed on d3 after injection. The degree of photoreceptor apoptosis was detected by HE staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling ( TUNEL ) and transmission electron microscope ( TEM ) in the right eyes. The mRNA expressions of rhodopsin and recoverin were detected different time after injection by Western blot and immunohistochemical method in the left eyes.
RESULTS:The dissolution of photoreceptor nucleus and apoptosis body were first perceived at 12h by TEM; most of cells at outer nuclear layer were presented positive reaction. The apoptotic index reached peak ( 29. 7% ±2.3%) at 24h which was coincided with the observation of TEM. The results of immunohistochemistry displayed that rhodopsin and recoverin were on a declining curve with time extension. Furthermore, the results of Western blot indicated that rhodopsin had dramatic decline at 6h after injection (P<0. 05), and extremely significant difference comparing to control group after 12h ( P<0. 01 ); while recoverin dramatic declined at 12h, and extremely significant difference after 24h (P<0. 01).
CONCLUSION:60mg/kg MNU intraperitoneally injection one - time may specifically induce photoreceptor apoptosis, The mechanism of down - regulation of rhodopsin and recoverin may be related to the selected apoptosis of photoreceptors.

Objective To investigate the influence of exogenous basic fibroblast growth factor(bFGF) on glial fibrillary acidic protein(GFAP) expression in neonatal rats with hypoxic-ischemic brain damage(HIBD).Methods Eighty HIBD models of neonatal Wistar rats were made by shearing right arteria carotis communis and then breathing 8% oxygen and 92% nitrogen for 2 hours.The models were divided into 2 groups randomly: trial group of bFGF and control group of normal sodium.The other 40 rats were taken into the sham(ope-)ration group.Expressions of GFAP were examined with immunohistochemical staining and image quantitative analysis at the 4~(th),7~(th),10~(th),17~(th) and 24~(th) days after operation.Results The expression of GFAP in hippocampal CA1 region of rats in sham operation group reached peak at the 7~(th) day after(ope-)ration.The expression of GFAP in control group increased and reached peak at the 10~(th) day after(ope-)ration,which GFAP-positive cells mainly appeared in hippocampal CA1 region and CA3 region.The expression of GFAP in hippocampal CA1 region of rats of trial group was higher than those of sham operation group and control group,which reached peak at the 10~(th) day after the operation,there was significant difference in 3 groups at the 4~(th),10~(th) and 17~(th) days after operation(all P

Objective To study the effect of ribozyme mediated inhibition on telomerase activity and cell(proliferation) of GBC-SD cells.Methods According to the hTR template two wings region sequence,the hammerhead ribozyme gene sequence was synthesized in vitro.Then,it was engineered into the eukaryon(expression) vector pTriEx-4.Lipofectamine was used to transfect the carcinoma of gallbladder.The inhibition of telomerase activity and cell proliferation in GBC-SD cell was observed,and compared with control group.(Results) Compared to control group,ribozyme can significantly inhibit telomerase activity and cell proliferation of GBC-SD cell.After ribozyme action,the cell ratio of G_0/G_1 markedly increased,and the cell ratio of S phase markedly decreased.Cellular cleavage and proliferation was markedly inhibited.The apoptosis rate was 17.10% and 31.01% on day10 and day13 after ribozyme action.Conclusions Ribozyme can inhibit(telomerase) activity of GBC-SD cells effectively and induce cell apoptosis.