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Objective:To analysis the Mimotopes of the peptide mimics to PGN using online softwares.Methods: Mimotopes of PGN were screened from 12-mers linear phage display peptide library by using anti-PGN McAb and the antigenicity of selected clones was identified by ELISA.The B cell epitopes,T cell epitopes of ′GRWxHxVxWAGL′ were estimated by DNAstar and online softwares.Results: 16 phage clones that bound with anti-PGN McAb were screened from 12-mers linear phage display peptide library.Among these positive clones,phage clones No.39 shared the conserved sequence:WxHx……AGL found in previous clone No.31(ATWxHxLxSAGL),which provoked an effective protective immunity against infection with S.aureus.To enhance the stability of the conformation as well as adding biotin on the N-ter-minal as a tag,the sequences ′39′ were redesigned and synthesized by adding S(serine)A(alanine) and GG(glycine)on the C-terminal of origin sequence(named SP39).Next,we estimated or predicted antigenic epitopes,T cell epitopes and scores binding to MHC of these peptides by using DNASTAR and online softwares(http://bio.dfci.harvard.edu/Tools/antigenic.pl,www.syfpeithi.de,http://www.darrenflower.info/mhcpred),indicating that SP39 contains sites bound both mice and human MHC.The sequences ′WxHxVxW-′ may be antigenic epitope as SP39,which contains a T cell epitope.Our results showed that both SP39 could bind to both anti-PGN McAb and a polyclonal antibody against S.aureus.Moreover PGN could inhibit the binding of SP39 to the anti-PGN McAb.These data indicated that SP39 mimic to epitopes on PGN.Conclusion: SP39(GRWxHxVxWAGLAGGS) probably display the mimotopes of PGN.
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Tinnitus is recognized as a refractory disease, which is common in clinic, and always treated from the liver and kidney. We treat tinnitus mainly by heart based on syndrome differentiation. It often works well when the leftpulse is abnormal. Referring to ancient literature, we find that the heart is related to ears by meridians, and the kidney and heart govern ears. The normal function of heart and kidney means normal hearing. The heart secretes atrial natriuretic polypeptins (ANP), whose receptor is widely distributed in ears and can increase the cochlear blood flow. In this paper we explore the relationship between the heart and the ears so as to provide theories of treatment by heart for tinnitus.
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Objective:To evaluate immune response of murine peritoneal macrophage challenging by methicillin-resistant S.aureus(MRSA)after pretreatment with Pam3Csk4(TLR2 agonist).Methods: Murine peritoneal macrophage was pretreated with Pam3Csk4(1 μg/ml).Following pretreatment 12 h later,heat-killed MRSA( HK-MRSA) was added and incubated for another 2 or 6 hours.The protein and mRNA level of TNF-α, IL-6 and IL-1 were determined by ELISA and Q-PCR, respectively.To estimate phagocytosis of macrophage,HK-MRSA/MSSA labeled with FITC( FITC-HK-MRSA/MSSA) were added to well and incubated for 30 min.After washing 5 times with PBS,intracellular FITC-HK-MRSA was detected by flow cytometry.To estimate antimicrobal activity of macrophage,live MRSA and MSSA were added to well and incubated at indication time,the CFU of s.aureus was estimated via a 10-fold serial dilution on agar media.cDNA was further quantitative assessed using primers for mouse FCR-Ⅰ,FCR-Ⅲ,CR-1,CR-3,iNOS and LL37 by Q-PCR .Results: Compared with saline-pretreated cell, the protein and mRNA level of TNF-α, IL-6 and IL-1 were markely reduced, respectively.However, both the phagocytosis and antimicrobal activity to S.aureus were significantly increased in macrophages pretreated with Pam3Csk4.Further study found that the macrophages had higher FCR-Ⅰ,FCR-Ⅲ,CR-1,CR-3,iNOS and LL37 expression at 6 h and 12 h post-stimulation Pam3Csk4.Conclusion: The results suggest that Pam3Csk4 could activate murine antimicrobal activity of peritoneal macrophage challenging by methicillin-resistant Saureus via increasing opsonophagocytosis in depended antibodies, complements manners.The results suggest Pam3Csk4 probably be a novel immunotherapy candidate against MRSA.
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Objective:To study the moxibustion at Guanyuan(CV 4)and Tianshu(ST 25)on intestinal dvsbacteriosis in rats.Methods:Fifty Wistar rats,clearing grade,were randomly divided into 5 groups,a normal group,a model group,a medical group,and a Guanyuan(CV 4)group and a Tianshu(ST 25)group,with 10 rats in each group.The rats were treated with Lincomycin Hydrochloride via intragastric administration for developing models.The model rats were treated with medicine and moxibustion respectively.0.1 g fresh rat feces in each group were cultured on the selective culture medium of bifidobacterium species(BS),lactobacillus(LBS),enterobactefiaceae(EB)and enterococcus(EC).The growth and quantity of the bacterial colony were tested by biochemical identification tubes and turbidimetry.Results:Moxibustion at Guanyuan(CV 4)increased BS and LBS while moxibustion at Tianshu(ST 25)improved EB and EC.Conclusion:Moxibustion at Front-Mu points Of different body parts selectively regulated advantaged probiotics for treating intestinal dysbactenosis.
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Objective To observe the changes of renal protein expression after electroacupuncture (EA) of "Taixi" (KI 3) of the Kidney Meridian of Foot-Shaoyin in healthy rats so as to explore the relationship between KI 3 and the Kidney. Methods Twelve adult male Wistar rats were randomly divided into normal control group (n=6) and EA group (n=6). EA (2 Hz/100 Hz,2-4 mA) was applied to bilateral KI 3 for 20 min,once a day for 7 days. Under anesthesia,the rats were perfused transcardiacally with ice saline and their kidneys removed for extracting the total proteins and assaying the differentially expressed proteins with two-dimensional gel electrophoresis (2-DE),Image MasterTM 2 D Platinum Software and matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) MALDI-TOF-MS,respectively. Results Image MasterTM 2 D Platinum Software analysis showed that after EA,9 protein points of the kidney tissues expressed differentially above 3-fold in comparison with control group were upregulated,2 of which were identified to be NAD-dependent isocitrate dehydrogenase and Quinone reductase. No downregulated differentially expressed proteins were found in the kidney tissues. Conclusion EA of "Taixi" (KI 3) can increase the expression of NAD-dependent isocitrate dehydrogenase and Quinone reductase in the kidney tissue,suggesting an increase of energy metabolism after EA and a close correlation between the KI 3 (Source-point) and the Kidney.
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Objective To observe the effect on body distribution tendency of paclitaxel liposome in the lung cancer rats after acupuncture of Feishu,Taiyuan acupoints in order to develop the new path for the target study of anticancer drugs.Method The paclitaxel liposome was labeled by technetium(Tc).Four methods including tail intravenous injection of paclitaxel liposome merely and the combination of tail intravenous injection of paclitaxel liposome with acupuncture of Feishu,Taiyuan(concerned acupoint) and Shenmen(unconcerned acupoint) acupoints respectively were carried on the intervention to the lung cancer rat models(18 in each group).The rats were sentenced to death at 60,90,120 minutes(6 at each time point) respectively after drug injection,and the drug content of paclitaxel liposome in the lung,liver,kidney organs were measureed by the ?-radiate immunity counting implement respectively.Result Acupuncturing at Feishu and Taiyuan can lead to the drug content increment of paclitaxel liposome in the lung organization of the lung cancer rats in all or parts of time points.On the contrary,acupuncturing at Shenmen can not.Conclusion Acupuncturing Feishu and Taiyuan can make tropism effect on the disposition of paclitaxel liposome in vivo in lung cancer rats.Feishu is stronger than Taiyuan on the drug tropism of the lung.
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Objective Adopting acupuncture associating with TAX to treat mice with Lewis lung cancer,to investigate the anti-cancer effect and its mechanism,and affirm their cooperating effect. Methods Lewis lung cancer model in mice was established and treated with acupuncture,medication and the combination of acupuncture and medication separately. The growth state of the mice,change and growth curve of tumour,change of tumour-weight and tumour-curbed rate were observed. Results Acupuncture,TAX and the combination of acupuncture and TAX all can curb the tumour,and the combination was the best. Conclusion The combination of acupuncture and TAX has a marked anti-cancer effect and can improve growth state of the mice. The effect of combining method is better than single therapy.