OBJECTIVE: To optimize the macroporous absorbing resin which were of best action in adsorption and desorption on the active components in Naomaitong granules. METHODS: UV spectrophotometry and HPLC was employed to determine the adsorbability and desorption capacity of different macroporous absorbing resins on total anthraquinones, total ginsenosides, total alkaloids and Puerarin. RESULTS: There were differences in adsorption and desorption capacity on active components in Naomaitong granules among different macroporous absorbing resins. Considering the general adsorbability and desorption capacity of different macroporous absorbing resins, AB-8 turned out to be of the best purification effect on Naomaitong granules. CONCLUSION: The results serve as a theoretical basis for the production of Naomaitong granules.

The envelope gene gp85 of ev/J,a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian ieukosis virus. This report described expression of ev/J envelope gene gp85 derived from commercial meat-type chicken using the Invitrogen Bac-to-Bac baculovirus expression system. The antigenicity and immunoreactivity of the recombinant endogenous gp85 gene product (SU) were analyzed by indirect immunofluorescence, Western blot, indirect and blocking Enzyme-Linked ImmunoSorbent Assay (ELISA) using JE9 monoclonal antibody (MAb) against the envelope protein of ALV-J (ADOL-4817), positive mouse antiserum against the ev/J gp85 SU and sera from chicken naturally infected with ALV-J. The results showed that the ev/J gp85 SU can bind specifically to JE9 MAb and antiserum from chicken naturally infected with ALV-J, and the binding reactivity between exogenous ALV-J gp85 SU and natural positive chicken serum against exogenous ALV-J can be blocked by positive mouse serum against the ev/J gp85 SU. It is concluded that recombinant endogenous gp85 gene product (SU) has close immunological relatedness to the envelope protein of exogenous ALV-J (ADOL-4817 and IMC<,10200> strain).

Objective To investigate the effect of different subtypes of fractionated doses on multidrug resistance in ovarian cancer cells.Methods The human ovarian cancer cell lines SKOV3 and its drug-resistant subtype SKVCR were divided into four groups i.e., sham-irradiated, single dose (10 Gy),fractionated dose (2 Gy × 5 ) and multi-fractionated dose (1 Gy × 2 × 5).Cell sensitivity to vincristine(VCR),etoposide ( VP-16),pirarubicin (THP) and cisplatin (DDP) was measured by MTT assay.Western blot was applied to detect the expression of P-gp after irradiation.Results The doubling time of SKVCR was about 1.8-fold of that of SKOV3 cells.P-gp was expressed in SKVCR but not in SKOV3.IC50 values of SKVCR were higher than those of SKOV3.To SKOV3 cells,single dose irradiation decreased cell sensitivity to THP and DDP and fractionated irradiation decreased cell sensitivity to VCR,THP and VP-16.Multi-fractionated irradiation decreased cell sensitivity to VP-16.In SKVCR cells,all these irradiation treatments increased cell sensitivity to VCR and VP-16 but not to DDP.In addition,single and fractionated irradiation decreased P-gp expression in SKVCR cells.Conclusions Single,fractionated and multi-fractionated radiation induced chemotherapy resistance in SKOV3 cells,while reversed drug resistance to VCR and VP-16 in SKVCR cells.

Objective To investigate the effects and mechanisms of sulindac, a non-selective cyclooxygenase(COX)inhibitor, on the proliferation and apoptosis of colorectal cancer cell line HT-29.Methods HT-29 cells were treated with sulindac. MTT assay and flow cytometry were used to measure the proliferation and apoptosis respectively. The laser scanning microscope(LSM)and the fluorescence microscope were used to observe apoptosis of the cells, and the flow cytometry (FCM)analysis was used to observe the cell apoptosis and cell cycle. Results Sulindac inhibited the cells proliferation and induced apoptosis in a dose-and time-dependent manner. With the TUNEL staining and fluorescence microscope, we found that the apoptosis cell became brown. After the Annexin V/PI staining, we observed that the membrane of apoptosis cells became green with LSM; the nucleosidase became red or crocus. FCM showed that sulindac promoted apoptosis of the cells, made the stage of G0/G1 ceils significantly reduced. Conclusions Our results showed that sulindac may inhibit the proliferation and induced the apoptosis of colon cancer cell HT-29,and the mechanism may probably be related to cell cycle arrest.

Objective:To discuss the screening results and clinical characteristics of children of Miao and Dong nationalities with mediterranean anemia in ethnic minority areas of Qiandongnan State of Guizhou Province,and to clarify the differences of the mediterranean anemia among different minorities.Methods:A total of 1 623 children of Miao and Dong nationalities with mediterranean anemia in minority areas of Qiandongnan State were selected by multistage stratified random sampling method;quantitative analysis of HbA2 and HbF was used to screen the selected children with mediterranean anemia initially;phenol chloroform extraction method was applied to extract the DNA from the children with mediterranean anemia;ASO/RDB-PCR reverse dot blot hybridization method was used to analyze the gene characteristics of the children with mediterranean anemia.Results:A total of 1 623 children of Miao and Dong nationalities were selected as the subjects.Among 938 children with Miao nationality,there were 18 children with positive α-mediterranean anemia and 36 children with positive β-mediterranean anemia,and the positive detection rate was 1.92%.Among 685 children with Tong nationality,there were 13 children with positive α-mediterranean anemia and 24 children with positive β-mediterranean anemia,and the positive detection rate was 3.50%.The detection rates of composite of α-and β-mediterranean anemia in the children of Miao nationality and Tong nationality were 1.49% and 4.61%.There was no significant difference in the detection rates of different kinds of mealiterranean anemia between two nationalities (P<0.05).The major gene mutations in α-mediterranean anemia were——SEA/-αα and-α3.7,and the major gene mutations in β-mediterranean anemia were CD17/N and CD14-15/N,while the major gene types of the composite of α-and β-mediterranean anemia were——SEA/β41-42 and——SEA/β17.There was no difference in the positive rates of major gene types of different kinds of mediter ranean anemia between two nationalities(P<0.05).Conclusion:There is no difference in the positive rate of children of Miao and Dong nationalities with mediterranean anemia in minority areas of Qiandongnan State.CD17/N,——SEA/-αα and ——SEA/β41-42 are the major gene types of α-,β-,and αβ-mediterranean anemia,respectively.

Objective To explore the expression and significance of hsa-miR-181a (miR-181a) in can-ceration progression of endometrial carcinoma. Methods A total of 75 formalin-fixed paraffin-embedded tissue specimens were studied in this study , of which , 13 were normal endometrium , 18 were endometrial hyperplasia , 44 were endometrial carcinoma. After total RNA had been extracted , real-time PCR was applied to detect the ex-pression level of miR-181a in endometrial tissue in each group. Results miR-181a expression in formalin-fixed paraffin-embedded tissue specimens can be detected. Expression of miR-181a in endometrial carcinoma was high-er than that in endometrial hyperplasia , its expression in endometrial hyperplasia was also higher than that in normal endometrium, and the difference was statistically significant (P < 0.05). The expression of miR-181a in endometrial carcinoma was associated with FIGO stages (P < 0.05). Conclusion The up-regulation of miR-181a expression in women with endometrial carcinoma may play the role of oncogenes. Abnormal expression of miR-181a is probably associated with the occurrence and development of endometrial carcinoma.

AIM: To study the purification of Naomaitong Granules(Radix et Rhizoma rhei,Rhizoma chuanxiong,Radix Puerariae lobatae.etc) by macroporous absorption resin. METHODS: Naomaitong Granule was purificated by macroporous absorption resin AB-8.UV spectrothotometry was used to determine the contents of total anthraquinones,total ginsenosides,total alkaloids,and the content of puerarin was determined by HPLC.The technic of purification was optimized according to the content above. RESULTS: The optimized technological conditions consisted of eoncentranon of original sample 120 mg/mL,the diameter and height was in proportion of 1∶10,the ratio of maximum adsorption to resin volume was 1∶6,water elution was 2B multiple of resin volume,8B multiple of resin volume 50% alcohol was the elution. CONCLUSION: AB-8 macroporous absorption resin can be used to purify Naomaitong Granules.

AIM:To establish the HPLC fingerprint of Compound Naomaitong effective parts,and to study the correlation analysis between fingerprint peaks and the effective fraction and its relevant herbs. METHODS:The chromatographic fingerprints of the effective fraction and the relevant fractions of its herbs were configured by HPLC/PDAD analysis. The relative deviation of retention time was utilized as indices to evaluate the correlation, the wavelength was set at 203 nm. RESULTS:The fingerprint of Compound Naomaitong effective parts was established and 36 copossessing fingerprint peaks were indicated. The assignment results of 14 peaks effective parts of fraction were indicated. CONCLUSION:The quality of Compound Naomaitong effective parts can be controlled by the HPLC fingerprint.

Objective Looking for the chemical components which have positive correlations with anti-inflammatiory activity of Flos Lonicerea to set up pharmacodynamic-fingerprint. Methods To gain the fingerprints of different parts by HPLC and to detect the pharmacological activities of anti-inflammation, then the correlationship between chemical components and pharmacological activities were detected by linear regression. Results Pharmacological activities of the methanol extracted part were best of all, so the fingerprint of the methanol part could represent the pharmacodynamic-fingerprint of Flos Lonicerea. Conclusion The fingerprint established by this way is more scientific and reasonable.

AIM:To establish the methods of determining 2,3,5,4′-tetrahydroxystilbene-2-O-?-D-glycoside,puerain;ginsengnoside Rg_1,ginsengnoside Rb_1,notoginsengnoside R_1,and salvianolic B in Jiangzhi Jianfei Tablets(Radix Polygoni multiflori,Radix Puerariae lobatae,Radix et Rhizoma Notoginseng,Radix et Rhizoma Salviae miltiorrhizae,etc) with HPLC. METHODS: All chromatogram columns used were ODS columns.The mobile phase of acetonitrile-wate(25(∶)75) was chosen for 2,3,5,4′-tetrahydroxystilbene-2-?-D-glycoside.The flow rate was 1.0 mL/min.The detection wavelength was at 320 nm.The mobile phase of methanol-water(25(∶)75) was chosen for puerain.The flow rate was 1.0 mL/min.The detection wavelength was at 250 nm.The mobile phase of acetonitrile-water was chosen for ginsengnoside Rg_1,ginsengnoside Rb_1 and notoginsengnoside R_1.The flow rate was 1.0 mL/min.The detection wavelength was at 203 nm.The mobile phase of methanol-acetonitrile-methane acid-water(30(∶)10(∶)1(∶)59) was chosen for salvianolic B.The flow rate was 1.0 mL/min.The detection wavelength was 286 nm. RESULTS: The linear rangers were 0.476 ?g-4.760 ?g(r=0.999 6) for 2,3,5,4′-tetrahydroxystilbene-2-?-D-glycoside,0.168 96 ?g-1.689 6 ?g(r=0.999 8) for puerain,0.724 ?g-7.240 ?g(r=0.999 8) for ginsengnoside Rg_1,0.728 ?g-7.280 ?g(r=0.999 9) for ginsengnoside Rb_1,0.23 ?g-2.30 ?g(r=0.999 8) for notoginsengnoside R_1,0.306 ?g-3.060 ?g(r=0.999 9) for salvianolic B.Their average recoveries were 99.43%(RSD=1.08%,n=6),99.43%(RSD=1.66%,n=6),101.17%(RSD=2.08%,n=6),and 102.39%(RSD=2.60%,n=6),respectively. CONCLUSION: These methods of determining components in four kinds of herbs are simple,accurate,reproducible and can be used for quality control of Jiangzhi Jianfei Tablets.