Molecular imprinting technique (MIT) involves the synthesis of polymer in the presence of a template to produce complementary binding sites in terms of its size, shape and functional group orientation. Such kind of polymer possesses specific recognition ability towards its template molecule. Despite the rapid development of MIT over the years, the majority of the template molecules that have been studied are small molecules, while molecular imprinting of proteins remains a significant yet challenging task due to their large size, structural flexibility and complex conformation. This review, we summarized the research findings over the past years, and discussed the nano-reinforcing materials used to prepare molecular imprinting of proteins and the perspective of these nano-reinforcing materials.
Binding Sites ; Molecular Conformation ; Molecular Imprinting ; Nanostructures ; chemistry ; Polymers ; chemistry ; Proteins ; chemistry

OBJECTIVETo study the killing effects of the liposome-mediated thymidine kinase (TK)/ganciclovir (GCV) system on MG-63 osteosarcoma (OS) cells and its bystander effects.

METHODSLiposome-mediated TK gene transfected into MG-63 OS cells, the efficiency of transfection was analyzed by flow cytometry and observed under inverted fluorescence microscope. Non-transfected osteosarcoma MG-63 cells were divided into three groups,in the experimental group 1 transfected TK/GCV cells cultured in solutiona liquid mixture by supernatant by 1/10,1/7,1/5,1/2 ratio to original broth; in the experimental group 2 transfected cells cultured in solutiona liquid mixture of supernatant filtered through 0.22 microm filter by 1/10,1/7, 1/5, 1/2 ratio to original broth, in control group the transfection cells cultured in original culture solution. Cell growth inhibition rate and osteosarcoma cell sensitivity to TK/GCV system were measured by MTT assay in each group.

RESULTSThe TK gene was transfected into MG-63 OS cells successfully by liposome-mediated, flow cytometry instrument detection TK gene transfection cell transfection efficiency can reach 75.5%. Six days later the MTT assay showed that in the experimental group 1 inhibition rate of all concentration ratio of the mixed culture fluid were statistically significant as compared with the control group (P < 0.05), and in the experimental group 2 that of the 1/10 and 1/7 of concentration ratio of mixed culture medium was not statistically significant as compared with the control group (P > 0.05). TK gene transfected MG-63 cells increased with the the GCV concentration,the cell apoptosis rate increased.

CONCLUSIONThe experiment demonstrated that the MG-63 OS cells are sensitive to the liposome-mediated TK/GCV system and bystander effects are significant.

Apoptosis ; drug effects ; Bone Neoplasms ; enzymology ; genetics ; physiopathology ; Bystander Effect ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Ganciclovir ; toxicity ; Humans ; Osteosarcoma ; enzymology ; genetics ; physiopathology ; Thymidine Kinase ; genetics ; metabolism ; toxicity

Objective To explore the effect of nuclear factor-kappaB(NF-?B) on cardiac myocyte apoptosis and understand the molecular mechanism of decrease of heart function in septic shock rats.Methods Twenty Wistar rats were randomly divided into the septic shock group and the normal control group.The septic shock group(n=10) were fixed with anaesthesia and made into 1.5 cm slices just along the abdomen midline.Then the roots of appendices were returned to the belly cavity after being ligated and punctured 4 times with the No.14 pinhead.After that,the slices were sewn up layer by layer.After 12 hours,the rats were all sacrificed,the blood taken and the serum separated.In the end,the heart specimens of the septic shock group were set aside.Meanwhile,the normal control group were dealt with in the same way except that they were not subjected to cecal ligation and puncture.Then NF-?B protein levels in cardiac tissue and the index of cardiac myocyte apoptosis were measured.Results NF-?B protein levels in the septic shock group(9 cases were strong masculine gender,1 case was middle masculine gender)elevated significantly compared with the normal control group(8 cases were negative gender,2 cases were weak masculine gender) in cardiac tissue(P

Objective To evaluate the value of differential diagnosis on congenital biliary atresia(BA) and infantile hepatitis syndrome(IHS) by technetium-99m-diethyl-iminodiacetic acid(99Tcm-EHIDA)hepatobiliary scintigraphy with phenobarbitol sodium.Methods Fifty-eight infants with persistent jaundice were taken phenobarbitol sodium[5 mg/(kg?d)] ,bid ?7 d).Those who had not bowel and gallbladders radioactivity within 24 hours were diagnosed as the diagnostic criterion of BA.Those with bowel and gallbladders radioactivity within 24 hours were diagnosed as the diagnostic criterion of IHS,who then received 99Tcm-EHIDA hepatobiliary scintigraphy with single photon emission computed tomography(SPECT) instrument.The results of all children were analyzed and compared with pathology and clinical follow up results.Results 99Tcm-EHIDA hepatobiliary scintigraphy correctly diagnosed 24 infants with last diagnosis BA and 29 infants with last diagnosis IHS,5 neonates false positive in all 34 IHS patients.The sensitivity in the diagnosis of BA was 100%,the specificity and accuracy were 85.3% and 91.4%,restectively.The sensitivity was 85.3% in the diagnosis of IHS;the specificity and accuracy were 100% and 91.4%,respectively.Conclusions 99Tcm-EHIDA hepatobiliary scintigraphy with phenobarbitol sodium can accurately differentiate BA and HIS at early stage.

Fractional Flow Reserve, Myocardial ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods

Novel solid lipid nanoparticle (SLN) system is prepared with Compritol ATO 888 and tricaprylic glyceride. DSC, XRD, SAXS and NMR are employed to study the novel carrier property and microstructure. When the peak melting point decreased from 70.8 degrees C to 61.4 degrees C, the enthalpy sharply decreased. It could be concluded that the regular crystal lattices in the novel carriers are broken out for the oil joined in them. Melting behavior is occurred at -17.7 degrees C while novel SLN is composed of oil and solid lipid mixture from the DSC measurement. Most alpha phase and least beta' phase are in the nano carrier system whether drug loading or not from the XRD investigation. There is only 0.1 nm change of long space among the novel SLN made of mixture and the lipid matrix and traditional SLN; therefore, it is impossible of the oil molecular insert into the solid glyceride structure. Since the different melting behavior (DSC measurements) and molecular move state (NMR investigations), two lipid matrix are still in two state of liquid and solid lipid in the novel SLN carrier. Presume the microstructure of the novel SLN prepared by our experiment would be that liquid oil has formed superfine nano accommodation encapsulated with solid lipid, but the whole particle is still in nano size range.
Calorimetry, Differential Scanning ; Caprylates ; chemistry ; Diterpenes ; administration & dosage ; chemistry ; Drug Carriers ; chemistry ; Drug Delivery Systems ; Epoxy Compounds ; administration & dosage ; chemistry ; Fatty Acids ; chemistry ; Magnetic Resonance Spectroscopy ; Nanoparticles ; Particle Size ; Phenanthrenes ; administration & dosage ; chemistry ; Triglycerides ; chemistry ; X-Ray Diffraction

OBJECTIVETo observe the immune effect of DNA vaccines encoding mutated HBV pre-c/c gene (VE2,VE4) in mice.

METHODSThree kinds of plasmid VEC(DNA vaccines encoding HBV pre-c/c gene), VE2 and VE4 were injected into the thigh muscles of different group of BALB/c mice.Blood and splenocytes from mice were isolated at 4 weeks after immunization. We also have mouse groups immunized with three of these plasmid combined with IFN-gamma gene plasmids. The anti-HBc and anti-HBe antibody in peripheral blood in mice were detected by enzyme linked immunosorbent assay (ELISA), antigen-specific cell immune responses were detected by CTL test and enzyme linked immunospot assay(ELISpot).

RESULTSWe found that anti-HBe titers of VE2 and VE4 immunizing groups are higher than VEC group (P < 0.05). We also observed that VE2 and VE4 could induce stronger antigen-specific immune responses than VEC and when combined with IFN-gamma plasmid,the antigen-specific immune responses are stronger than those without combination immunization in mice (P < 0.05).

CONCLUSIONSThe DNA vaccine VE2 and VE4 could induces stronger antigen-specific immune responses than VEC, and when combined with IFN-gamma plasmid,the antigen-specific immune responses are improved in mice.

Animals ; Female ; Hepatitis B ; prevention & control ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B Vaccines ; administration & dosage ; Hepatitis B virus ; genetics ; Immunization ; Male ; Mice ; Mice, Inbred BALB C ; Mutation ; Vaccines, DNA ; administration & dosage ; genetics ; immunology

OBJECTIVETo investigate the effects of oxidized low-density lipoprotein receptor 1 (LOX-1) on secretion of adhesive molecules mediated by ox-LDL in human umbilical endothelial cells (HUVECs).

METHODSHUVECs with different concentration of ox-LDL (0, 10, 20, 50, 100 microg/ml) were incubated for 24 h, or HUVECs were pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for another 24 h. Expression of LOX-1 was determined by realtime RT-PCR and Western blot. mRNA and protein of ICAM-1, VCAM-1 and E-selectin were examined by RT-PCR and Western blot respectively.

RESULTSIncubation of HUVECs with ox-LDL (10-100 microg/ml) enhanced the expressions of LOX-1, ICAM-1 and E-selectin in a concentration-dependent manner (P < 0.01). On the contrary, ox-LDL did not affect the expression of VCAM-1 by HUVECs. The expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL were reduced in HUVECs pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for 24 h. This showed that both poly (I) and carrageenan obviously decreased the expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL.

CONCLUSIONox-LDL may upregulate the expression of LOX-1, ICAM-1 and E-selectin, and LOX-1 blocker may partly inhibit this upregulation. The results suggest that the expression of inflammatory molecules induced by ox-LDL in HUVECs is mediated by LOX-1.

Cell Adhesion ; Cell Adhesion Molecules ; Cells, Cultured ; E-Selectin ; metabolism ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipoproteins, LDL ; biosynthesis ; RNA, Messenger ; metabolism ; Receptors, Oxidized LDL ; metabolism ; Scavenger Receptors, Class E ; metabolism ; Umbilical Veins ; cytology ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo demonstrate molecular characterization of a newly isolated enterovirus.

METHODSVirus were isolated from patient with unknown-causing disease and tested by reverse transcription-polymerase chain reaction (RT-PCR) and 5'3'RACE (rapid amplification of cDNA ends, RACE), in an attempt to obtain the sequence of this newly isolated enterovirus.

RESULTSSequence analysis showed that this newly isolated enterovirus shared 83%-94% nucleotide identity and 91%-100% amino acid identity with enterovirus 89. Phylogenetic analysis indicated that it was probably a new subtype of enterovirus 89.

CONCLUSIONThis newly isolated enterovirus in the stool specimen from patient has the same serotype with enterovirus 89, but it was probably a new subtype of enterovirus 89.

Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Enterovirus ; classification ; genetics ; isolation & purification ; Feces ; virology ; Genome, Viral ; genetics ; Humans ; RNA, Viral ; genetics ; Sequence Analysis, DNA

BSA liposomes were prepared with approximately 100 nm mean particle size under rather gentle experiment conditions, and two-colorimetric coomassie brilliant blue protein was employed to measure the free drug in the entrapped efficiency (EE%) determination of BSA liposomes. Gel filtration was used to measure the EE%, and several Sephadex gels were examined by the separation of liposomes and free drug. To determine the free drug, three methods were compared on two-colorimetric UV spectrophotography, Bradford and two-colorimetric coomassie brilliant blue, separately. Two-colorimetric coomassie brilliant blue process increased the accuracy and improved the sensitivity of the assay about 20-fold comparing with the Bradford method. Two-colorimetric coomassie brilliant blue assay appeared to be more sensitive and showed broader dynamic range to measure the free BSA in the EE% determination of BSA liposome.
Colorimetry ; Drug Carriers ; Drug Compounding ; Electrophoresis, Gel, Two-Dimensional ; Liposomes ; Particle Size ; Rosaniline Dyes ; Serum Albumin, Bovine ; administration & dosage ; analysis ; Spectrophotometry, Ultraviolet