Objective To investigate the effects and the gene expression of mitochondria on arsenic trioxide-induced apoptosis of acute promyelocytic leukemia (APL) NB4 cells. Methods NB4 cells were treated with As2O3. Fluorescent microscopy and flow cytometry were used and mitochondria membrane potential detection and RT-PCR were performed to observe the NB4 cell apoptosis, growth inhibitory effect and mitochondrial transmembrane potentials. Mitochondria genome primers were designed, and the expression of mitochondria genome at gene and protein levels was studied. Results Induced by As2O3, the NB4 cells showed typical morphological changes of apoptosis with a significant growth inhibitory effect. The ratios of NB4 cells apoptosis were 5.02%, 6.40%, 28.40% and 33.34%, respectively, when treated with As2O3 in concentrations of 0.5?mol/L, 1?mol/L, 2?mol/L and 3?mol/L. When treated with As2O3 in 0.5?mol/L, 1?mol/L, 2?mol/L, 4?mol/L and 8?mol/L at 48h, the mitochondria potential of NB4 cells was decreased by 12.8%, 21.6%, 66.9%, 83.7% and 83.8%, respectively. After the NB4 cell apoptosis was induced by As2O3, RT-PCR assay was used to detect the expression of 13 genes of mitochondria genome. The expression of COX2 gene was down-regulated in this process, while no change was found in the expression of other 12 genes. Conclusions As2O3 has shown to exert a significant effect on promoting apoptosis and growth inhibition on APL cells. The apoptotic effect induced by As2O3 on NB4 cells is closely related to a decrease of mitochondrial membrane potential. The expression change in the mitochondria gene COX2 is involved in the As2O3-induced apoptosis of NB4 cells.

Objective To observe the efficacy of arsenic trioxide(ATO) combined all trans retinoic acid (ATRA) versus cytara‐bine (Ara‐C) combined ATRA in the treatment of acute promyelocytic leukemia(APL) .Methods We enrolled 65 patients in our department during the period between January 2002 and August 2008 ,and they were randomly assigned to receive ATRA combined ATO (treatment group ,n= 27) or ATRA combined DA ,HA ,NA which were major of Ara‐C (control group ,n= 38) .Then observe the differences of between the two groups ,such as complete remission(CR) ,the time to complete remission ,overall survival(OS) ,e‐vent free survival(EFS) ,the 5 years disease free survival (DFS) and adverse reactions .Results The CR rate of treatment group (ATRA + ATO) and control group (chemotherapy + ATRA) was 81 .48% and 68 .42% ,respectively ,and the time to complete re‐mission was (28 .50 ± 3 .97)d and (30 .56 ± 2 .39)d ,respectively ,showed that there was no statistical difference between the two groups ( P > 0 .05 ) .The 5 years DFS of the CR patients in the two groups was 51 .9% (ATRA + ATO ) and 50 .0%(Chemotherapy + ATRA) ,respectively ,showed that there was no statistical difference between the two groups(P > 0 .05) .The 5 years EFS of the CR patients in the two groups was 48 .1% and 39 .5% ,respectively ,showed that there was no statistical difference between the two groups(P> 0 .05) .The 5 years DFS of the patients in the two groups was 55 .6% and 67 .6% ,respectively ,showed that there was no statistical difference between the two groups(P > 0 .05) .Bone marrow suppression in the treatment group was significantly lower than in the control group(P< 0 .05) .Conclusion ATRA + ATO can prolong the CR rate ,OS ,EFS and 5 years EFS of newly diagnosed APL patients .ATRA combined with chemotherapy has similar efficacy ,ATRA + ATO has lower bone marrow suppression than the ATRA combined with chemotherapy ,thus may reduce the risk of early death .

AIM: To analyze the chemical composition and their relative contents of Fuyinjie Compound Huangsong Lotion. METHODS: A capillary column HP-5 MS was used. The column temperature was controlled by a program and the MS analysis was performed with EI and quadrupole mass analyzer. The chemical composition was identified by NIST98 searching and mass spectra comparing, and their relative contents were determined by using normalization method of chromatographic peak areas. RESULTS: There were 39 components found and 31 composed of more than 90% separated components were identified. CONCLUSION: The GC-MS is a simple, rapid and sensitive method for the chemical composition analysis of Fuyinjie Compound Huangsong Lotion.

Objective Gene microarray technology was used to investigate the differential gene expression of apoptosis related genes in NB4 cells induced by arsenic trioxide. Methods The databases of Evntrez and Human IPI were searched with "apoptosis or apoptotic" as the key words, and 1384 apoptosis related genes were found after the redundant genes were eliminated by chromosomal localization. The probes of these genes were designed using OligoArray 2.0, and then analyzed by BLAST. All the probes were immobilized on the glass slide, which were used as oligonucleotide microarray. After NB4 cells were treated with 2umol/L As2O3 for 48h, the total RNA were extracted. cDNAs of control group and test group were fluorescently labeled with Cy3 and Cy5, respectively, by RT-PCR. The fluorescent samples were hybridized with an oligonucleotide microarray containing 1384 apoptosis related genes to search for the differentially expressed genes in the cells with or without As2O3 treatment. The hybridization signals were scanned by oligonucleotide microarray, and then the fluorescent intensity of Cy3 and Cy5 and the ratio of two fluoresceins were analyzed using certain software. Then the differential expressed genes were analyzed after As2O3 treatment, in which the most distinctly differential expressed genes were chosen as targets, and through PCR amplification and gel electrophoresis the above genes were verified. Results There are 4 genes up-regulated and 12 genes down-regulated in expression in NB4 cells after 48h treatment with 2umol/L As2O3, which were in accordance with the results of RT-PCR and oligonucleotide microarray. Conclusion Differential gene expression in NB4 cells was induced by As2O3 treatment. These differentially expressed genes, with relation to signal transduction, transcription regulation, cell cycles, oxidation response, protein translation and cell differentiation, may play an important role in NB4 cell apoptosis.

Object To develop a method for the determination of flavonoids from the tender stem and leaf of Ampelosis grossedentata (Hand. Mazz.) W. T. Wang, and for its quality control. Methods Differential spectrophotometry was used at the wavelength of (318?1) nm, while taking dihydromyricetin as index. Results The regression equation, within the range of the control concentration in 4 576～22 88 ?g/mL, was A=29 56 C+0 005 5, with a good linearity (r=0 999 8) between the absorbance and concentration. The average recovery rate was 100 8% and RSD was 1 9% (n=5). Conclusion The method is simple, reliable and easy to operate and suitable for the quality control of Chinese medicinal materials with a better reproducibility.

Physiology experiment can effectively stimulate the enthusiasm of the students to explore the scientific truth by providing the opportunity to observe real physiological phenomena,analyze,discuss and solve practical problems.Lab-based learning (LBL) and theory teaching have complementary advantages,promoting each other and can maximize the use of teaching resources.During the process of physiological LBL,training of students' thinking ability by creating problem situation,inspiration and guidance,carrying out exploring experiments can develop students' intelligence,promote innovation,improve the teaching effect.

Objective:To compare the effects between electroacupuncture (EA) at Chize (LU 5, the He-Sea point of the Lung Meridian) and Shangjuxu (ST 37, the lower He-Sea point of the large intestine) in rats with ulcerative colitis (UC) on the variations of mesenteric microcirculation and vasoactive intestinal peptide (VIP) in the colon, lung, and hypothalamus. The relative specificity of acupoints was also explored.
Methods: A total of 28 male Wistar rats were randomized into a normal group, a model group, a Chize (LU 5) group and a Shangjuxu (ST 37) group, 7 rats in each group. The UC model was established by enema with acetic acid. Since the third day after modeling, rats in the Chize (LU 5) group and Shangjuxu (ST 37) group respectively received EA at Chize (LU 5) and Shangjuxu (ST 37), 15 min each time for successive 7 d. The variations of mesenteric microvascular calibers and blood flow status were observed by a microcirculation microscopic tester; VIP in the colon, lung and hypothalamus was measured by radioimmunoassay.
Results:Compared with the normal group, the mesenteric microvascular calibers were significantly expanded in the model group (P<0.05); there was no significant difference between the model group and Chize (LU 5) group (P>0.05); compared with the model group and Chize (LU 5) group, the calibers were obviously shrunk in Shangjuxu (ST 37) group (P<0.05). The four groups showed no significant inter-group differences in comparing blood flow status (P>0.05). The colonic VIP levels in the model group and Chize (LU 5) group were significantly higher than that in the normal group (P<0.01,P<0.05); the VIP level in Shangjuxu (ST 37) group was markedly lower than that in the model group (P<0.01). There were no significant differences among the four groups in comparing VIP level in lung and hypothalamus (P>0.05).
Conclusion:The effects of Chize (LU 5) and Shangjuxu (ST 37) were different in treating UC. Shangjuxu (ST 37) showed a more significant efficacy in down-regulating VIP in the colon and regulating mesenteric microcirculation, while the effects of Chize (LU 5) were not obvious.

AIM:To establish an HPLC-ELSD method for determing dulcitol and astragaloside in Compound Fufangteng Capsules(Radix et Rhizoma Ginseng rubra,Radix Astragali,Herba Euonymi,etc).METHOD:Chro-matographic conditions included Lichrospher 5-NH_2 column(250 mm?4.6 mm,5 ?m)and the mobile phase consisting of a mixture of acetonitrile-water(91 ∶9).The flow rate of mobile phase was 1.0 mL/min.The column tempe-rature was at 28 ℃.Detector:PL-ELS2100 ELSD(Eva:65 ℃,Ned:50 ℃,gas flow:0.9 L/min).RESULTS:The linear ranges of dulcitol and astragaloside were 2.91-29.1 ?g(r= 0.999 8)and 1.03-10.3 ?g(r= 0.999 1),respectively.The average recoveries of dulcitol and astragaloside were 99.24% and 103.17% with corresponding RSD of 2.6 % and 1.8%(n=6),respectively.CONCLUSION:The method is steady and with good repeatability,and can be used to determine the content of dulcitol and astragaloside in Compound Fufangteng Capsules.

AIM: To establish a RP-HPLC method to determine the fingerprint of Compound Fufangteng Mixture (Radix Ginseng Rubra, Radix Astragel. etc). METHODS: Chromatographic conditions included Shim-pack CLC ODS column and the mobile phase composed of a mixture of acetonitrile-water (gradient elution), detection wavelength at 203nm. The mobile phase flow rate was 1.0mL?min -1. RESULTS: 19 peaks existed on the HPLC fingerprint of Compound Fufangteng Mixture. CONCLUSION: The method can provide more information for the quality control of Compound Fufangteng Mixture.

OBJECTIVE:To establish a method for the simultaneous determination of syringinand and pedunculoside in Zhuang medicine Yuyang powder. METHODS:HPLC was performed on the column of Agilent ODS with mobile phase of acetonitrile-water (gradient elution)at a flow rate of 1 ml/min,the detection wavelength was 210 nm,the column temperature was 30℃,and the in-jection volume was 5μl. RESULTS:The linear range was 0.71-3.55μg for syringin(r=0.999 7)and 1.62-8.10μg for pedunculoside (r=0.999 8), respectively;RSDs of precision, stability and reproducibility tests were lower than 3%;recoveries were 100.8%-104.9%(RSD=1.7%,n=6) and 96.0%-100.8%(RSD=2.2%,n=6). CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous determination of syringinand and pedunculoside in Zhuang medicine Yuyang powder.