Development of a novel insulin receptor (IR) antagonist that exhibits anti-breast tumor activity.

Many reports have indicated that the insulin receptor (IR) causes tumorigenesis and the development of breast cancer. It has been considered a potential target for treating IR-related tumors. Traditionally, there are two categories of insulin receptor (IR) antagonists, they are small molecule antagonists and anti-IR antibodies. Here, we describe a new method (anti-idiotypic antibody strategy) for the development of IR antagonist. Hybridoma technology was employed to design and identify a series of anti-idiotypic antibodies against insulin. After repeated screening and identification, an anti-idiotypic antibody against IR (AK98) was obtained. Analysis through competitive ELISA and competitive receptor binding indicated that AK98 mimicked the receptor binding epitope of insulin. The interaction between AK98 and IR was determined using indirect immunofluorescence, immunoelectron microscopy, and Immunoprecipitation-Western (IP-WB). Further research using a tumor cell model revealed that AK98 inhibited insulin-IR binding and IR-mediated intracellular signaling pathways. Conclusively, the main purpose of this paper is that we proposed a new method (anti-idiotypic antibody strategy) to develop the insulin receptor (IR) antagonist (AK98), and a series of experiments showed that the anti-idiotypic antibody (AK98) exhibited good antagonistic activity against IR. This work suggests that the anti-idiotypic antibody may be a potential strategy to develop IR antagonists that can be used in treating breast cancer.

Efficacy and safety research of multimodal analgesia with different doses of nalbuphine combined with flurbiprofen in intravenous patient -controlled analgesia after thoracotomy / 中国基层医药

Objective To assess the efficacy and safety of multimodal analgesia with different doses of nalbuphine combined with flurbiprofen on patients who received intravenous patient -controlled analgesia(PCIA) after thoracotomy.Methods Sixty patients underwent thoracotomy,ASA Ⅰ -Ⅱ,18 -65 years old,who underwent postoperative PCIA,were randomly divideded into three groups according to the digital table,nalbuphine 60 mg group (N60 group),nalbuphine 80 mg group(N80 group)and nalbuphine 100 mg group (N100 group),20 cases in each group.All patients were given 150mg flurbiprofen,a total of 100mL.PCIA solution:the background dose was 2mL/h, PCIA dose of 0.5mL,locking time of 15min.10min before surgery,each patient was intravenously given flurbiprofen 50mg,given a loading dose of 0.1mL/kg when closed chest.All patients were followed up for 48h.The incidence of adverse reactions such as vital signs,number of times,visual analog scale(VAS)score,sedation score,nausea and vomiting were recorded.Results There were no significant differences in the age,gender,body mass index and surgery duration among the three groups(all P >0.05).The vital signs were stable within 48h after operation.The VAS scores of N60 group were higher than the other two groups(N80 group:t =7.94,6.35,6.49,5.21,5.63,all P =0.00;N100 group:t =8.41,9.10,5.80,8.07,8.18,all P =0.00)at 4,6,8,24 and 48h after operation(all P 0.05).The effective /actual compression ratio of PCIA of N80 group and N100 group were significantly higher than that of N60 group (t =7.30,8.35,all P 0.05;group N100:χ2 =3.14,0.23,1.03,all P >0.05).Conclusion Postoperative PCIA with nalbu-phine (80 mg)combined with flurbiprofen(150 mg)has significant analgesic effect and lower costs.

Research Review of Functional Mechanism of Treatment by Way of Pasting TCM on Acupoints / 中国中医药信息杂志

Treatment by way of pasting TCM on acupoints has a long history and the clinical efficacy is determined. In this article, based on the concept of TCM wholism and meridian theory, the mechanism of acupoint pasting therapy was expounded from the aspects of medicine effects on local stimulation of the body, meridian conduction, medicine transdermal absorption. Combining with the research hot spots of modern medicine transdermal delivery system, metabolomics, and efficacy material, this article proposed a new study mode of systems biology of TCM.

Genetic quality analysis of 24 domestic inbred mouse strains by microsatellite DNA / 中国比较医学杂志

Objective To analyze the genetic quality of 24 domestic inbred strains mice using microsatellite loci panel.Methods Previously selected 30 microsatellite loci of mouse with high polymorphism and more allele numbers were used to synthesize corresponding fluorescently-labeled primers.Then the genomic DNA samples of each mouse were amplified by PCR and the products were analyzed by STR scanning to genotype the inbred strains of mice.Results Out of the 24 inbred strains, 15 inbred strains showed the same genotype within one strain at 30 loci.Among different strains, microsatellite loci indicated polymorphism which could be used to distinguish different strains.However, the rest 9 strains demonstrated polymorphism within strains.Conclusions Our stuoly provides a useful microsatellite panel to detect genetic quality of inbred mice and distinguish different strains with the optimized microsatellite loci.

Research Progress in Animal Models of IBS-D Disease Combined with Liver Stagnation and Spleen Deficiency / 中国中医药信息杂志

At present, TCM treatment of diarrhea-predominant irritable bowel syndrome (IBS-D) is based on the combination of disease differentiation and syndrome differentiation. Therefore, the establishment of IBS-D of liver stagnation and spleen deficiency syndrome combined with animal model as a combination of traditional Chinese and Western medicine innovation theory has become increasingly concerned about, and gradually become a new direction for the development of TCM experimental animal model. This article reviewed the research progress in IBS-D liver and spleen deficiency syndrome in recent years, discussed the establishment of IBS-D liver stagnation and spleen deficiency animal model and research ideas for the treatment of IBS-D, and provided references for mechanism research of TCM treatment for IBS-D and research and development of new medicine.

Cloning,Expression and Immunization of The Hypoxanthine-guanine Phosphoribosyltransferase for Schistosoma japonicum Chinese Strain / 生物化学与生物物理进展

A 1 270 bp full-length cDNA fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the 3′ and 5′ ends of the incomplete expression sequence tag (EST) of hypoxanthine-guanine phosphoribosyltransferase of Schistosoma japonicum (SjHGPRT) were amplified by the anchored PCR with 2 pairs of primer that were designed according to the published incomplete SjHGPRT EST and the sequence of multiclone sites of library ?gt11 vector. Sequence analysis indicated that this fragment, with an identity of 82% to hypoxanthine-guanine phosphoribosyltransferase of Schistosoma mansoni (SmHGPRT), contained a complete open reading frame(ORF). The deduced amino acid sequence showed 83% identity to that of SmHGPRT. This fragment was cloned into the prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE revealed that M of the recombinant protein was about 28 ku. Western-blot analysis showed that the recombinant protein was recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Mice vaccinated with recombinant protein revealed significant worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs of the female worms reduction percentage, compared with the controls. Taken together, the SjHGPRT full-length cDNA can be cloned and expressed in E.coli as a recombinant protein that elicited immunity against the challenge infection with Schistosoma japonicum, indicating its potential as a partial protection vaccine candidate.

Cloning, Expression and Immunization of The Hypoxanthine-guanine Phosphoribosyltransferase for Schistosoma japonicum Chinese Strain / 生物化学与生物物理进展

A 1 270 bp full-length cDNA fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the '3' and 5' ends of the incomplete expression sequence tag (EST) of hypoxanthine-guanine phosphoribosyltransferase of Schistosoma japonicum (SjHGPRT) were amplified by the anchored PCR with 2 pairs of primer that were designed according to the published incomplete SjHGPRT EST and the sequence of multiclone sites of library λgt1 1 vector. Sequence analysis indicated that this fragment, with an identity of 82% to hypoxanthine-guanine phosphoribosyltransferase ofSchistosoma mansoni (SmHGPRT), contained a complete open reading frame(ORF). The deduced amino acid sequence showed 83% identity to that of SmHGPRT. This fragment was cloned into the prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE revealed that M of the recombinant protein was about 28 ku. Western-blot analysis showed that the recombinant protein was recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Mice vaccinated with recombinant protein revealed significant worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs of the female worms reduction percentage, compared with the controls. Taken together, the SjHGPRT full-length cDNA can be cloned and expressed in E. coli as a recombinant protein that elicited immunity against the challenge infection with Schistosoma japonicum, indicating its potential as a partia1 protection vaccine candidate.