Adjuvants have been used as critical components of vaccine.They were used to stabilize the antigens and potentiate the immune responses.Dose sparing is also of interests in recent years to be applied in potential pandemic situations or to lower the costs of vaccines.With the emerging nanotechnology in many aspects including the use in biomedical applications,the application of nanomaterials as vaccine adjuvants also attracted a lot of attentions in recent years.With favorable biological properties,such as well-defined and well-formed nanoparticles,biocompatibility,biodegradability and ability to induce humoral immunity and cellular immunity simultaneously,calcium phosphate nanoparticles (CaP) have the potentials to be developed as an immune potentiator to be used as vaccine adjuvants.Here,we reviewed the basic properties of CaP and their applications as vaccine adjuvants with antigens of different modalities such as inactivated virus,protein and naked DNAs.The mechanism of the adjuvanting effects is briefly described.Further the development of CaP as vaccine adjuvants with some designed surface modification could lead to next generation vaccine adjuvants with improved immune potentiating properties and safety profiles.

Objective To induce the the 17th condon antonymous mutagenesis of salivary histatin 5(HRP5) cDNA,express the mutant and hrp5 in Pichia pastoris,and to study the effects of mutation on expression.Methods According to the Pichia pastoris' codon bias,two pairs of primers(H1 and H2,H3 and H4) were designed.H1 and H2,H3 and H4 have complementary 3' end,and the EcoR I site was added to the 5' end of H1 and H3,Sal I site to H2,H4.The cDNA of hrp5 and hrp5' was generated with PCR by H1 and H2,H3 and H4,respectively.The secrete vector pPICZ?-A,hrp5 and hrp5' were digested with EcoR I +Sal I,linkede by T4 DNA ligase and transformed to E.coli TOP10 comptetent cell,positive colonies were screened on LB plates with Zeocin.The recombinant plasmids pPICZ?-A-hrp5 and pPICZ?-A-hrp5' identified by digestion and DNA sequencing were amplified largely,linearized by Sac I and transformed to GS115 comptetent cell by electroporation,positive colonies were screened on YEPD plates with Zeocin,the recombinant GS115 were confirmed by PCR,cultured and induced expression by methanol.The amount and anticandidal activity of the expressed products was compared with synthetic HRP5.Results Both hrp5 and hrp5' were integrated into the genome of GS115 and expressed successfully,the anticandidal activity of the recombinant HRP5 and HRP5' was identical with synthetic HRP5,the amount of expressed HRP5 and HRP5' was 4?mol/L and 5?mol/L,respectively.Conclusion Both the recombinant HRP5 and HRP5' showed better anticandidal activity.The amount of expressed prodcts increased 25% by substituting Asn for Lys17 without changing anticandidal activity.

BACKGROUND: The existing electrocardiogram(ECG)measurement strongly depends on medical professionals and inefficient high-intensity,or relies on automatic identification method which is not accurately enough.Thus,this is difficult to meet high-speed testing,accurate results and ease application for common people.OBJECTIVE: To develop a new method that was simple and efficient to apply and very easy to learn.METHODS: Algorithms were programmed and test software was developed by delphi7.0.ECG was drawn on screen.The apex,the starting point and the ending point as well as the J-point of each ECG wave were clicked by mouse or stylus.Then the wave parameters and an initial diagnosis could be quickly obtained by test software.RESULTS AND CONCLUSION: The parameters of ECG waveform such as wave height,wave time,PR interval,ST segment,QT segment,PP/RR time,cardiac electrical axis and so on could be accurately measured,and heart rate,heart rhythm and the deflection of cardiac electrical axis could be diagnosed correctly.The method was simple to learn and easy to imply,and it was also efficient,quick and accurate.Thus,it could greatly improve the efficiency of measurement and analysis for specialists,and could meet application requirements of general medicals and ordinary people.

The effects of melittin on the activities of Na+-K+-ATPase and glucose-6-phosphate dehydrogenase (G-6-PD) which are on the membrane of red blood cell (RBC) are chosen as the index of this study. The possible target sites of these effects through enzyme activity determination by spectrophotometry are investigated, and the hemolytic process and the activity change of these two types of enzymes on the RBC membrane are discussed. The results show that the main mode of melittin inhibition to the activity of enzymes on the RBC membrane is the coexistence of adhesion/insertion form and free-state form, and the effect of the former is more stronger than the latter. The K+ binding site of Na+-K+-ATPase is one of the target sites of melittin. The membrane-insertion process of melittin synchronizes with the action of melittin on this enzyme. Melittin slowly inhibits the catalysis of G-6-PD through the action on G-6-P and NADP, and the extent in which melittin forms tetramers isclosely related to the enzyme activity. EDTA inhibits the aggregation of melittin, and interferes with its action on G-6-P. The biochemical mechanisms of melittin effects on the substrate G-6-P and the coenzyme NADP are similar, and the inhibition of melittin is not related to the structure of G-6-PD.

Objective To achieve the fusion expression of the entire human beta-defensin-3(hBD-3) gene. Method We synthesized two oligonucleotide primers accor ding to the codon preference of Escherichia coli. The gene was cloned into p GEX -4T-2 to establish the pGEX-4T-2-hBD-3 as the fusion expression vector by PCR. Transformed into E.coli strain DH5α, the express vector was induced an d ex pressed by IPTG. The fusion protein GST-hBD-3 was obtained by repeated cycles of freezing and thawing, cut by thrombin to attain the recombinant hBD-3 protei n. Result The result of the antibacterial peptide agarose diffu sion assay shows the antibacterial activity of the rhBD-3 against the S.aureu s exists, and it reaches 0.843U. Conclusion The fusion expr ession of the hBD-3 gene is successful.

Background:As the empirical studies on human body are restricted extremely,the establishment and selection of suitable animal models are important for researches on ulcerative colitis( UC ). Aims:To compare the symptoms and colonic pathology of rat models with experimental colitis induced by dextran sulfate sodium( DSS ) and trinitrobenzene sulfonic acid( TNBS),so as to provide a reference for selecting animal models in UC-related studies. Methods:Drinking 4% DSS freely for 7 days or intrarectal administration of single dose 100 mg/kg TNBS-50% ethanol were used to establish experimental colitis model in Sprague-Dawley rats. The disease activity index( DAI)was assessed dynamically during the course of experiment. The whole colon was removed in batches for measurements of colonic damage score and activity of myeloperoxidase(MPO)at different time points. Results:The DAI score reached the peak at the 7th day and the 2nd day in DSS group and TNBS group,respectively,and decreased gradually afterwards. Six and one deaths occurred during the experimental course in DSS and TNBS groups,respectively. In DSS group,the duration of inflammation was short,the colonic injury was moderate and recovered after drug withdrawal. At the 18th day,the colonic damage score and MPO activity was 0. 25 ± 0. 50 and(0. 80 ± 0. 33)U/g,respectively,and no significant differences were seen between DSS group and normal control group. In TNBS group,the duration of inflammation was longer and the colonic injury was more severe. At the 21st day,the colonic damage score and MPO activity was 3. 60 ± 0. 55 and( 1. 60 ± 0. 39 ) U/g, respectively,and chronic inflammation was observed histologically. Conclusions:Both DSS and TNBS can induce experimental colitis model in rats. The course of TNBS-induced colitis model presents a transformation of acute to chronic inflammation,and may be more suitable for treatment-related studies of UC.

OBJECTIVETo prepare the polyclonal antibody against methyl-accepting chemotaxis signal transduction protein (MCP) of Helicobacter hepaticus (H.hepaticus).

METHODSThe recombinant plasmid pET22b+/MCP was transformed into E.coli BL2l(DE3) to express the fusion protein His-rhMCP under the induction of IPTG. The fusion protein was purified and the antibody was obtained by immunizing rabbits. The titer of the polyclonal antibody was tested by indirect ELISA, and the specificity of the antibody was identified based on Western blotting using the prepared cell surface proteins (CSPs) of the bacteria.

RESULTSThe fusion protein was successfully expressed, and the titer of the antibody reached 1:32 000. Western blotting indicated that the antibody could specifically bind to CSPs and His-rhMCP.

CONCLUSIONThe antibody with a high titer and specificity was prepared to facilitate further study of the pathogenicity and epidemiology of H.hepaticus in human.

Animals ; Antibodies, Bacterial ; biosynthesis ; genetics ; Antibody Specificity ; Bacterial Proteins ; immunology ; Helicobacter hepaticus ; immunology ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Signal Transduction

OBJECTIVETo observe the effect of gut protease activity on visceral hypersensitivity in rats with acute restraint stress.

METHODSSprague-Dawley rats were given 30, 100 or 300 mg/kg camostat mesilate (CM), a protease inhibitor, or saline intragastrically 30 min before acute restraint stress induced by wrapping the fore shoulders, upper forelimbs and thoracic trunk for 2 h. Visceral perception of the rats was quantified as the visceral motor response with an electromyography, and the rectal mucosa and feces protease activity and spinal c-fos expression were measured.

RESULTSCM dose-dependently reduced visceral sensitization elicited by rectal distension, but these doses did not completely inhibit stress-induced visceral sensitization. In normal rats, c-fos expression was found mainly in the superal spinal cord dorsal horn, and after the administration the CM, c-fos-positive cells decreased significantly in all dose groups (P<0.05). In 30 mg/kg CM group, fecal and rectal mucosal protease activity significantly decreased as compared with that in the stress group (P<0.05), and as CM dose increased to 100 and 300 mg/kg, the protease activity decreased even further (P<0.01).

CONCLUSIONThe gut protease is involved in acute stress-induced visceral hypersensitivity, and CM can lower the visceral sensitivity and spinal c-fos expression in rats.

Animals ; Gabexate ; analogs & derivatives ; pharmacology ; Protease Inhibitors ; pharmacology ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats ; Rats, Sprague-Dawley ; Restraint, Physical ; Spinal Cord ; metabolism ; Stress, Physiological

The fatality rate of traumatic cardiac arrest (TCA) is extremely high, and it is very different from that of non-traumatic cardiac arrest (NTCA) in resuscitation strategy. Only when the standard resuscitation process is combined with rapid treatment of various reversible causes can the mortality rate of patients be decreased. In this paper, the key factors leading to TCA are reviewed, such as hypovolemic shock, asphyxia, tension pneumothorax, pericardial tamponade, crush syndrome, craniocerebral injury, cerebral hernia, and the control measures are elaborated respectively, so as to provide references for clinical treatment of patients with severe trauma, and reduce TCA incidence and mortality.

Objective: To investigate the dosimetry differences of target and OARs of an integrated design of fields in IMRT and the mainstream IMRT technique for post-radical mastectomy. Methods: A total of 41 patients with post-radical mastectomy who received IMRT were eligible, the conventional fixing two-degrade collimator and the integrated IMRT fields were designed respectively. The dosimetry parameters of target and OARs, monitor units and delivery time of both plans were compared. Results: The dose distribution for targets and OARs of both plans met clinical requirements. The dosimetry parameters of target of both plans showed no statistically significant difference (P>0.05). Compared with the conventional technique, the integrated IMRT plans showed significant advantages, the ipsilateral lung V₅ decreased by 9.7%(t=2.407, P<0.05), V₁₀ 11.2%(t=2.160, P<0.05), V₂₀ 17.3%(t=2.465, P<0.05), V₃₀ 13.4%(t=2.119, P<0.05), D_mean 13.8%(t=2.258, P<0.05). And the heart V₃₀ decreased by 28.4%(t=2.589, P<0.05), D_mean 23.2%(t=2.409, P<0.05). The dosimetric differences of other OARS were not statistically significant(P>0.05). Conclusions The new method can effectively reduce exposed volume and exposed dose of ipsilateral lung and heart without affecting the target dose coverage. The method has universal applicability to patients with post-radical mastectomy who received IMRT, with important clinical significance.