The relationship of insulin dosage during transition from continuous subcutaneous insulin infusion(CSII)to subcutaneous injections of premixed human insulin in 197 type 2 diabetic patients was analyzed. Positive correlation(r=0.60,P

Re-evaluation of bioequivalence of generic drugs is one of the key research focus currently. As a means to ensure consistency of the therapeutic effectiveness of drug products, clinical bioequivalence has been widely accepted as a gold standard test. In vitro dissolution testing based on the theory of the BCS is the best alternative to in vivo bioequivalence study. In this article, the conventional dissolution method and flow-through cell method were used to investigate the dissolution profiles of domestic amoxicillin capsules in different dissolution media, and the absorption behavior of the drugs with different release rates (t85% = 15-180 min) in the gastrointestinal tract was predicted by Gastro Plus. The flow-through cell method was thought better to reflect the release characteristics in vivo, and amoxicillin capsules with regard to the release rates up to 45 min (t85% = 45 min) were having a satisfied bioequivalence with the oral solution according to the C(max) and AUC. Although two different dissolution profiles of domestic amoxicillin capsules were found by flow-through cell methods, prediction results revealed that domestic capsules were probably bioequivalent to each other.
Amoxicillin ; pharmacokinetics ; Capsules ; Computer Simulation ; Gastrointestinal Tract ; Humans ; Software ; Solubility ; Therapeutic Equivalency

Objective: To screen candidate antigen for therapeutic HBV vaccine with a novel adjuvant DC-Chol. Methods: BALB/c mice were injected with DC-Chol liposome and HBsAg prepared from CHO and Yeast respectively. One week later, IL-4, IL-2, IFN-?were measured by ELISA or ELISPOT. Results: The levels of IL-2, IFN-?of HBsAg from Yeast with DC-Chol liposome were 20 and 119 times higher respectively than those of HBsAg from CHO with DC-Chol liposome. ELISPOT assay showed that the counts of spot-forming cells of IL-4 and IFN-?of HBsAg from Yeast with DC-Chol liposome were 2.8 and 46.3 times higher respectively than those of HBsAg from Yeast with Al(OH)3. Conclusion: HBsAg prepared from Yeast together with DC-Chol liposome may be an appropriate candidate for therapeutic HBV vaccine .

Objective:Preparation and immune characteristic analysis of polyclonal antibody against hypervariable region protein of Taura syndrome virus major capsid protein VP 1 as a reference for studies on immunological diagnosis reagent.Methods:The recombinant vector pET-VP1 was transformed into E.coli BL21 for protein expression.Immunizing a New Zealand rabbit with purified VP1 protein,the titer of anti-VP1 serum was determined by Agar diffusion test and ELISA.Monoclonal phage specific binding to the purified VP1 protein was used for competitive inhibition test.Results: The VP1 protein was soluble and high expression in E.coli BL21.The biological activity titer of anti-VP1 serum reached 1∶26 ,1∶217 determined by Agar diffusion test and ELISA respectively.A litter binding activity of antiserum and VP 1 protein could be blocked by monoclonal phage , but would not affect the final positive result.Conclusion:High titer antibody Preparation of the VP 1 hypervariable region protein.The binding activity of the polyclonal antibody with VP1 protein was not affected by the mutations of VP 1 protein in minority areas ,so the antiserum could be used as immu-nological detection diagnosis agent.

Background Researches showed that wild-type p53(wtp53)and Rb94 genes inhibit the growth of retinoblastoma(RB),and these genes are involved in signal pathway in the induction and maintenance of cellular senescence.Thus the combination of two genes to inhibit growth of RB is concerned.Objective This study was to observe the inhibitory effect of the co-transfection of Rb94 and wtp53 gcnc into subretina on RB with ultrasound microbubble.Methods HXO-Rb44 suspension was subretinally injected to establish the RB model in 40 SPF female Balb/c nude mice.The RB models were randomized into model control group,wtp53 group,Rb94 group and wtp53+Rb94 group,and 0.1 ml relevant gene microvesicle suspension was injected via caudal vein in the different groups,but no any gene was used in the model control group.Seven days after modeling,followed by 0.5 W/cm2ultrasonic wave irradiated the eyeballs immediately for 4 seconds ×2 times and interrupted for 24 seconds.Eyeballs were extracted 7 days after gene transfection,and the expressions of wtp53 mRNA and Rb94 mRNA in tumor tissuc were detected by RT-PCR,and wtp53 and Rb94 protein in tumor tissue were assayed using Western blot.Immunochemistry was used to exam the VEGF expression,and TUNEL was used to evaluate the apoptosis of the tumor cells.Results The model successful rate after HXO-Rb44 suspension was 80％ (32/40)and obvious malformation cells were seen under the light microscope.In 7 days after gene transfection,no response band for wtp53 mRNA and Rb94 mRNA were found.The relative expression valuc of wtp53 mRNA was 0.65±0.07 in the wtp53 group,and that in wtp53+Rb94 group was 0.32±0.02,showing a significant difference between them (t =11.743,P =0.000).Rb94mRNA relative value was 0.42 ±0.03 in Rb94 group,and that in the wtp53 + Rb94 group was 0.23 ± 0.03,with a significant difference(t=5.041,P=0.001).The response bands of wtp53 and Rb94 proteins were seen in wtp53group,Rb94 group and wtp53+Rb94 group.Immunochemistry showed that the positive reactive intensity for VEGF in tumor tissue was obviously weaker in wtp53+Rb94 group than that in the wtp53 group,Rb94 group and model control group.Apoptotic index(Al) was 37.35±2.14 in the wtp53+Rb94 group,showing a significant increase in comparison with model control group (0.46 ± 0.05),wtp53 group (5.05 ± 0.80) and Rb94 group (6.43 ± 1.02) (t =-34.395,-28.206,-26.006,P＜0.01).Conclusions Ultrasound microvesicle enable double gene transfecting into RB tumor tissue,and Rb94 gene cooperation with wtp53 gene enhance the inhibitory effect on RB by promoting the apoptosis of RB cells.

Objective:To stablish of chemiluminescent Southern blot detection system for examining HBV DNA replication intermediates in HepG2.2.15,and analyse the inhibition of HBV replication with three kind of drugs with different targets.Methods:The HBV DNA replication intermediates were extracted and analyzed by Southern blot with HBV probe,which(pTHBV1047) was labelling with digoxigenin.The results of the hybridization were detected by chemiluminescent,and the condition of hybridization was optimized.After treated with lamivudine,Bay41-4109,?-Galcer in different concentration,the HBV DNA from the HepG2215 cells were detected with the system.Results:the sensitivity of the system was 1pg of pTHBV1047,and HBV specific positive signals was detected with the DNA from HepG2.2.15.The three kinds of drugs can inhibit the HBV replication obviously with chemiluminescent Southern blot detection system,the IC50 were 1.53?mol/L,0.41?mol/L,0.01?mol/L.Conclusion:The HBV replication intermediates from the cell of HepG2.2.15 can reflect the antiviral effect accurately with different targets drugs and this mothod would be used in the study of Chinese-midicine.

OBJECTIVETo study the effects of Volatile Oil of Radix Angelicae Sinensis (VOA) on experimental asthma in rat model based on abnormal immune functions of Treg cells.

METHODSAfter grouping, the asthmatic rats were developed through injecting OVA and AI(OH)3 for sensitization and then administering OVA aerosol for challenge, and the respiratory functions, asthmatic behaviors, IL-10 levels in bronchoalveolar lavage fluid (BALF) (ELISA) and Foxp3 expression (immunohistochemistry) in lung of asthmatic rats were observed.

RESULTSVOA at the doses of 40-160 mg/kg could improve the respiratory functions and the asthmatic behaviors, and upgrade IL-10 levels in BALF and Foxp3 expression in lung of asthmatic rats.

CONCLUSIONVOA has some effects of anti-asthma and one of the mechanisms is to improving the lower immune functions of Treg cells.

Angelica sinensis ; chemistry ; Animals ; Asthma ; drug therapy ; Bronchoalveolar Lavage Fluid ; chemistry ; Disease Models, Animal ; Forkhead Transcription Factors ; metabolism ; Interleukin-10 ; chemistry ; Lung ; metabolism ; Oils, Volatile ; pharmacology ; Plant Oils ; pharmacology ; Rats ; T-Lymphocytes, Regulatory ; cytology

Objective To establish a high performance liquid chroma-tography-tandam mass spectrometry ( HPLC-MS/MS) method for de-termining the concentration of sunitinib and N -desethyl sunitinib (SU12662) in human plasma.Methods Sunitinib, SU12662 and in-ternal standard sunitinib -D10 were extracted from plasma with one -step protein precipitation and the stereoselective analysis of sunitinib and SU12662 was achieved on an Agilent ZORBAX Extend C18 (2.1 mm ×75 mm ×3.5 μm) with gradient elution by a mobile phase consisting of wa-ter containing 0.1%formic acid and 95%acetonitrile.Electrospray ioni-zation source was applied, and multiple reaction monitoring mode was operated in the positive mode with the monitor ions at m/z 399.3→326.1 for sunitinib, m/z 371.3→283.1 for SU12662 and 409.3→326.2 for sunitinib-D10.Results The calibration curve for plasma sunitinib was linear in the range of 0.5-200 ng? mL-1 ( r=0.998 7) , the lower limi-tation of quantification was 0.5 ng? mL-1 , while the accuracy was ranged from 0.57% to 2.87%, the average recovery rate was ranged from 92.75%to 98.62%,and intra-and inter-day RSD were ranged from 0.91%to 1.92%and 5.40%to 7.87%, respectively.The calibration curve for plasma SU12662 was linear in the range of 0.25 -100 ng? mL-1 ( r =0.999 9 ) , while the accuracy was ranged from -2.08% to 2.80%, the average recovery rate was ranged from 96.23%to 101.12%, and intra-and inter-day RSD were ranged from 0.46%to 2.46%and 5.84%to 9.75%, respectively, the lower limitation of quantification was 0.25 ng? mL-1 .Conclusion The HPLC-MS/MS method was accurate, sensitive, specific, and could be used in the determination of sunitinib in plasma and the clinical study of its pharmacokinetics.

OBJECTIVETo investigate the relationship between the helicobacter pylori (HP) infection and the genetic instability of mitochondrial DNA (mtDNA) in human gastric adenocarcinoma epithelial cells (AGS).

METHODSAfter treated with extracts of HP11638 (CagA+, VacA+) or Hp11638 mutant strain (CagA+, VacA-), AGS cells were collected, and mitochondrial DNA was extracted and Cox-I, Cox-II, Cox-III, ATPase6, ATPase8 and Cytb genes and the D-Loop region were amplified by PCR and then sequenced.

RESULTSThe mutation rates of the mtDNA in AGS cells were correlated with the extracts of the two HP strains in a concentration- and time-dependent manner. But the mtDNA mutation rate in AGS cells treated with the HP11638 extract was higher than that treated with the Hp11638 mutant extract. Total of 616 mutations in D-Loop region were detected, including 489 point mutations, 81 insertions and 46 deletions. Among them, 70.9% (437/616) belonged to GC to AT and AT to GC transition. Seventeen out of 20 (85%) AGS cells treated with extract of HP had mutations in 303PolyC, 16184PolyC and 514CA regions of mtDNA D-Loop. No mutation was detected in Cox-I, Cox-II, Cox-III, ATPase6 and ATPase8 genes, three point mutations were found in the Cytb gene.

CONCLUSIONHP can cause the accumulation of mutations in mtDNA, in particular, in the D-Loop region, and the VacA participated in the process.

Antigens, Bacterial ; pharmacology ; Base Sequence ; DNA, Mitochondrial ; drug effects ; genetics ; Endothelial Cells ; drug effects ; pathology ; Helicobacter Infections ; complications ; Helicobacter pylori ; chemistry ; Humans ; Mutation ; Stomach ; pathology

OBJECTIVETo assess the value of fluorescence in situ hybridization (FISH) combined with chromosomal analysis for the detection of Robertsonian translocation type trisomy 21 in amniotic fluid cells.

METHODSAmniotic fluid samples from pregnant women requesting prenatal diagnosis were cultivated. Metaphase cells were prepared for G-banding karyotype analysis. For the 5 Robertsonian translocation type trisomy 21, interphase nuclei from amniotic fluid and parental peripheral blood cells were prepared for FISH analysis.

RESULTSIn 2 cases, analysis of parental peripheral blood cells showed normal karyotypes. FISH analysis of amniotic fluid cells indicated that one sample had two copies of chromosome 21, which has a 46, XY, rob(21;21)(q10;q10) karyotype, whilst another had trisomy 21 by FISH, which has a 46, XY, rob(14;21)(q10;q10) karyotype. For the remaining three samples, analysis of parental peripheral blood cells indicated that their karyotypes were 45, XX, rob(14;21)(q10;q10), 45, XX, rob(15;21)(q10;q10) and 45, XX, rob(21;22)(q10;q10), whilst the karyotypes of amniotic fluid cells were 46, XX, rob(14;21)(q10;q10), 46, XY, rob(15;21)(q10;q10) and 46, XX, rob(21;22)(q10;q10), respectively.

CONCLUSIONCombined FISH and chromosomal analysis is an efficient method for detecting non-homologous Robertsonian translocation type trisomy 21. However, FISH has limited ability to detect homologous Robertsonian translocation type trisomy 21.

Adult ; Down Syndrome ; diagnosis ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Karyotyping ; Pregnancy ; Prenatal Diagnosis ; Translocation, Genetic