Aim To investigate subcellular distribution and accumulation of ADR in the sensitive and multidrug-resistant HL-60 cells and its relation to multidrug resistance.Methods The subcellular distribution and accumulation of ADR were studied by confocal scanning laser microscope and flow cytometry.The effects of verapamil,BSO,brefeldin A and chloroquine on ADR distribution and accumulation in HL-60/ADR cells were also examined.Rhodamine123,NBD-ceramide and neutral red were used as fluorescent probes to stain the mitochondria,Golgi apparatus and lysosomes respectively were used to identify the subcellular compartments where ADR was sequestered.Results In drug-sensitive cell line HL-60,ADR fluorescence distributed evenly in the nucleus and cytoplasm,while in multidrug-resistant cell line HL-60/ADR,ADR fluorescence distributed in a punctated pattern in the cytoplasm and was reduced in the nucleus.The mode of ADR distribution in HL-60/ADR cells is highly similar to that of NBD-ceramide.BSO and brefeldin A,instead of verapamil and chloroquine could reverse the abnormal distribution and accumulation of ADR in HL-60/ADR cells.Conclusions The change of ADR distribution and reduction of ADR accumulation in multidrug-resistant cell line was involved in the mechanism of multidrug resistance.

The authors designed to separate, purify and determine the monosaccharide composition of the polysaccharide from Cordyceps militaris, and study its effect on reverse cholesterol transport in vivo by isotope tracing assay. Polysaccharides were separate and purify by ion exchange column Q-sepharose Fast Flow and size exclusion column Sephacryl S200HR; the molecular weight and monosaccharide composition of the polysaccharides were determined by high performance gel permeation chromatography and high performance liquid chromatography coming with pre-column derivation, respectively. Finally, three purified polysaccharides CMBW1, CMBW2 and CMYW1 were obtained, their total carbohydrate contents were 87%, 89%, 95%, respectively; their protein contents were 6.5%, 1.3%, 2.8%, respectively; their molecular weights were 772.1, 20.9, 13.2 kDa, respectively; CMBW1 was composed of mannose, glucosamine, rhamnose, glucuronic acid, glucose, galactose and arabinose with a molar ratio of 7.25: 0.17: 1.29: 0.23: 6.30: 11.08: 0.79; CMBW2 was composed of mannose, glucosamine, galactose and arabinose with a molar ratio of 2.40: 0.16: 2.92: 0.24; CMYW1 was composed of mannose, glucosamine, glucuronic acid and glucose with a molar ratio of 0.59: 0.57: 0.45: 25.61. Polysaccharide at 50 mg x kg(-1) could significantly improve the transport of 3H- cholesterol to blood and excretion from feces. All of the three purified polysaccharides CMBW1, CMBW2 and CMYW1 were heteropolysaccharide; and they could improve reverse cholesterol transport in vivo, the underlying mechanisms are being studied.
Animals ; Biological Transport ; drug effects ; Cholesterol ; metabolism ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Cordyceps ; chemistry ; Mice ; Monosaccharides ; analysis ; isolation & purification ; Polysaccharides ; chemistry ; isolation & purification ; pharmacology ; Tritium

Background and purpose: Primary signet ring cell carcinoma(SRCC) of the bladder is rarely diagnosed in the clinic. Few cases have been reported in the literature, so there was lack of understanding of the primary bladder SRCC in terms of diagnosis and treatment. Our study was to investigate the clinical features and treatment strategy for primary SRCC of the bladder and review the status of the disease along with the literature. Methods: 3 cases of primary bladder SRCC were studied, including clinical features, treatment, follow-up and their prognosis.The literature was reviewed. Results: All cases received ultrasound, computerized tomography, cystoscopy, biopsy and other related lab tests for diagnosis and differential diagnosis. Laparoscopic radical cystectomy and orthotopic ileal neobladders were performed in 2 cases, while the other case received laparoscopic radical cystectomy and ileal conduit diversion, Chemotherapy (cisplatin and 5-fluorouracil) was delivered in one case after surgery. One patient died at 6 months postoperatively because of multiple metastasis. The other 2 cases have been followed-up only for 8 and 12 months postoperatively, and no recurrence or metastasis have been observed. Conclusion: Primary SRCC of the bladder lacks distinctive clinical and imaging manifestations. The tumor grows very invasively. Radical cystcctomy is one of the optimal approaches for treatment of SRCC of bladder.

Background and purpose: The prognostic factors on survival for the patients with prostate carcinoma are still underdeterrnined. This study was to analyze the survival of three common treatment methods for prostate carcinoma and the prognostic factors on survival. Methods: 494 male patients who were diagnosed as prostate cancer were enrolled into the retrospective study. All of the data like age, stage, grade, PSA level, ALP, Hb and treatments were collected. Overall survival and disease specific survival rates for patients were analyzed by Kaplan-Meier method. Prognostic factors on disease specific survival were also analyzed by Log-rank test and Cox proportional hazards model. Results: Disease specific survival rates at 1, 3 and 5 year were 96.0%, 89.0% and 80.0% for all 494 patients, respectively. Disease specific survival rate at 3-year was 92.4% for brachytherapy, 100.0% for radical prostatectomy and 80.6% for hormonal therapy (P=0.008). Multivariate analysis by Cox model showed that stage, PSA level and age significantly impacted on disease specific survival. Conclusion: Brachytherapy and radical prostatectomy provides longer survival time than hormonal therapy for patients with prostate cancer. Clinical stage and PSA level and age of prostate cancer are independent factors impacting on survival significantly.

Objective To investigate association between human leukocyte antigen DQB1 (HLADQB1 ) gene polymorphisms and bronchial asthma among Mongolian and Han nationalities. Methods Sequence-specific primer polymerase chain reaction (PCR-SSP) was used to detect frequencies of HLA DQB1 genotypes and alleles in 50 cases of Han and 68 Mongolian asthmatic patients, and 50 Han and 54 Mongolian healthy controls, respectively. Difference in gene frequencies between the two nationalities was estimated by odds ratio (OR) and chi-square test. Results Frequency of the HLA-DQB1 0602 allele was significantly higher in Han patients with bronchial asthma than that in healthy Han nationality (OR = 6.163,P ＜0.01 ). Frequency of the HLA-DQB1 0603/0608 allele decreased in Mongolian asthmatic patients, as compared to that in healthy Mongolians ( OR = 0.199, P ＜ 0.05 ). Frequency of the HLA-DQB1 0301/4 allele was significantly higher in Mongolian asthmatic patients as compared to that in healthy Mongolians ( OR =2.074,P ＜0.05). Frequency of the HLA-DQB1 0301/4 allele was significantly higher in Mongolian than that in Han asthmatic patients ( OR = 2.482 ,P =0.05). Frequency of the HLA- DQB1 0602 allele was significantly higher in healthy Mongolians than that in healthy Han nationality ( OR = 3.341, P ＜ 0.05 ), in contrast, frequency of the HLA-DQB1 0402 allele was significantly lower in healthy Mongolians than that in healthy Han nationality ( OR = 0.209, P ＜ 0.05 ). Conclusions The HLA-DQB1 0603/0608 allele is possibly a protective gene and the HLA-DQB1 0301/4 allele a susceptible gene for bronchial asthma in Mongolians, and the HLA-DQB1 0602 allele is possibly a susceptible gene for bronchial asthma in Han nationlity.

Terminal restriction fragment length polymorphism (T_RFLP) analysis is a culture- independent approach for analyzing microbial community in environment. It bases on PCR technology, and its process includes DNA extraction of environmental samples, amplification of genes encoding the 16S rRNA, 18S rRNA or enzymes with fluorescently labeled primers, the restriction enzyme digestion of PCR products, cap- illary electrophoresis and the analysis of T_RFLP profile. It has been proved to be powerful applied on mi- crobial community in environment since developed in 1997. Currently, T_RFLP rarely applied in China, and it has no applications on microbial community analysis of nitrifying bacteria. In this article, the fundamental principle of this technique and the recent applications of T_RFLP on microbial community are summarized; in addition, it illustrates the confinements of conventional culture-dependent of nitrifying bacteria and the foreground of T_RFLP applying on microbial community structure analysis of nitrifying bacteria.

Objective:To study the correlation between HBV Pre-S1 antigen,HBeAg levels and HBV DNA copies,so as to assess the clinical value of Pre-S1 in detection of HBV replication.Methods:A total of 363 HBsAg-positive samples were col- lected.The levels of Pre-S1 antigen,HBeAg and HBV DNA copies were determined by ELISA,time-resolved immuno-fluores- cent method and real-time fluorescent quantitative PCR(FQ-PCR),respectively.The correlation between the determination re- sults was analyzed.Results:Pre-S1 antigen level was correlated with the level of HBeAg(X~2=94.4,P

Objective To explore the mechanism of percutaneous laser disc decompression (PLDD) for treating prolapse of intervertebral disc. Methods 15 rabbits were divided into 3 groups randomly: sham group, model group with PLDD treatment, model group without PLDD treatment. The nerve conduction velocity (NCV) of L6 nerve root and the activity of phospholipase A2 (PLA2) in the intervertebral disc of L5-6 were determined 2 weeks after the initial surgery. Results NCV in the group with PLDD was significantly faster than that in the group without PLDD (P<0.001); NCV in the group without PLDD was significantly lower than that in the sham group (P<0.001). The activity of PLA2 in the group with PLDD was significantly lower than that in the group without PLDD (P<0.001); The activity of PLA2 in the group without PLDD was significantly higher than that in the sham group (P<0.001). Conclusion The activity of PLA2 in the herniated discs is higher than that in normal discs, which result in NCV falls remarkably. The PLDD can reduces chemical factors such as PLA2.

Adult ; Aged ; Humans ; Male ; Middle Aged ; Mining ; Occupational Exposure ; Pneumoconiosis ; complications ; mortality ; Survival Analysis ; Tuberculosis ; complications ; mortality

Previous studies proposed that the synergistic effect of fibroblast growth factor-21 (FGF-21) and insulin may be due to the improvement of insulin sensitivity by FGF-21. However, there is no experimental evidence to support this. This study was designed to elucidate the mechanism of synergistic effect of FGF-21 and insulin in the regulation of glucose metabolism. The synergistic effect of FGF-21 and insulin on regulating glucose metabolism was demonstrated by investigating the glucose absorption rate by insulin resistance HepG2 cell model and the blood glucose chances in type 2 diabetic db/db mice after treatments with different concentrations of FGF-21 or/and insulin; The synergistic metabolism was revealed through detecting GLUT1 and GLUT4 transcription levels in the liver by real-time PCR method. The experimental results showed that FGF-21 and insulin have a synergistic effect on the regulation of glucose metabolism. The results of real-time PCR showed that the effective dose of FGF-21 could up-regulate the transcription level of GLUT1 in a dose-dependent manner, but had no effect on the transcription level of GLUT4. Insulin (4 u) alone could up-regulate the transcription level of GLUT4, yet had no effect on that of GLUT1. Ineffective dose 0.1 mg kg(-1) FGF-21 alone could not change the transcription level of GLUT1 or GLUT4. However, when the ineffective dose 0.1 mg x kg(-1) FGF-21 was used in combination with insulin (4 u) significantly increased the transcription levels of both GLUT1 and GLUT4, the transcription level of GLUT1 was similar to that treated with 5 time concentration of FGF-21 alone; the transcription level of GLUT4 is higher than that treated with insulin (4 u) alone. In summary, in the presence of FGF-21, insulin increases the sensitivity of FGF-21 through enhancing GLUT1 transcription. Vice versa, FGF-21 increases the sensitivity of insulin by stimulating GLUT4 transcription in the presence of insulin. FGF-21 and insulin exert a synergistic effect on glucose metabolism through mutual sensitization.
Animals ; Blood Glucose ; Diabetes Mellitus, Experimental ; metabolism ; Drug Synergism ; Fibroblast Growth Factors ; pharmacology ; Glucose ; metabolism ; Glucose Transporter Type 1 ; metabolism ; Glucose Transporter Type 4 ; metabolism ; Hep G2 Cells ; Humans ; Insulin ; pharmacology ; Insulin Resistance ; Liver ; metabolism ; Mice