Chemiluminescence behavior of CdTe-hydrogen peroxide enhanced by sodium hypochlorite and sensitized sensing of estrogens.

It has been found that sodium hypochlorite enhanced the chemiluminescence (CL) of the CdTe nanocrystal (NC)-hydrogen peroxide system and that estrogens inhibited these CL signals in alkaline solution. CL spectra were used to investigate the mechanism of the CL enhancement. On the basis of the inhibition, a flow-injection CL method has been established for determination of three natural estrogens.

A monoclonal antibody against a novel Sialomucin CD300LG.

CD300LG is a novel O-glycosylated member of the CD300 antigen-like family. Besides a classical mucin-like domain, it contains a V-type Ig domain. CD300LG binds lymphocyte L-selectin via its Ig domain and supports lymphocyte rolling via its mucin-like domain. The unique structure and function of CD300LG suggest it may play an important role in inflammation. For preparation of a monoclonal antibody (MAb) against human CD300LG, prokaryotic and eukaryotic expressing human CD300LG proteins were used as immunogen and detection antigen, respectively. One stable strain of hybridomas (3C7C5A6) was successfully established using the hybridoma technique. The Western blot and immunohistochemistry analyses demonstrated that the MAb was directed against human CD300LG with high specificity. This antibody could possibly facilitate studies on the pathomechanism of inflammation and may have the potential to be a means of effective anti-inflammation.

A novel monoclonal antibody against cannabinoid receptor 1.

The cannabinoid receptor 1 (CBR1) is being widely investigated because of its specific structure and functions compared with other cannabinoid receptors. In this study, we immunized BALB/c mice with synthesized human CBR1 polypeptide and obtained a novel monoclonal antibody (MAb) against human CBR1. Analysis through enzyme-linked immunosorbent assay (ELISA), spot-ELISA, Western blot, and immunohistochemistry revealed that the MAb was specifically against recombinant human CBR1 protein, and its subtype and affinity constant (Kaff) were IgG2b/k and 7.85 × 10(8) M/L, respectively. Using this MAb we found that CBR1 is expressed on HL-7702 cells and lipid tissue, raising the possibility that the CBR1 may take a role in glucose and lipid metabolism. Thus, this antibody might facilitate studies for pathophysiology of diseases associated with glucose and lipid metabolism abnormality.

Self-assembly of copper and cobalt complexes with hierarchical size and catalytic properties for hydroxylation of phenol.

A feasible and effective self-assembly method to synthesize different scale coordination polymers in highly dilute solution (from nanocrystals to microcrystals and to bulk crystals) without any blocking agent has been described. The growth of crystalline particles was controlled by removing the particles at different reaction times to interrupt the growth at the desired size. The nano and microscale particles show better catalytic conversions and selectivities in the hydroxylation of phenols than the bulk crystals.

catena-Poly[[diaqua-zinc(II)]-µ-piper-azine-1,4-diacetato-κN,O:N,O].

The asymmetric unit of the title compound, [Zn(C(8)H(12)N(2)O(4))(H(2)O)(2)](n), contains a Zn(II) ion residing on an inversion center, half of a centrosymmetric piperazine-1,4-diacetate ligand (L) and a water mol-ecule. The Zn(II) ion is trans-coordinated by two N,O-bidentate L ligands and by two water mol-ecules in a distorted octa-hedral geometry. In the crystal structure, inter-molecular O-H⋯O hydrogen bonds link polymeric chains into a three-dimensional supra-molecular structure.

[Oxalylbis(aza-nedi-yl)]bis-{[amino-(2-pyrid-yl)methyl-ene]ammonium}.

The title compound, C(14)H(16)N(8)O(2) (2+)·2ClO(4) (-), was prepared by reaction of bis-[amino-(2-pyrid-yl)methyl-ene]oxalohydrazide with perchloric acid. The mol-ecular symmetry is C(i) and thus the asymmetric unit comprises one half-mol-ecule. The dihedral angle between the aromatic ring and the plane of the oxamide group is 70.8 (3)°. The perchlorate anions and the cations are connected by inter-molecular N-H⋯O hydrogen bonds.

trans-Diaqua-bis[(E)-3-(dimethyl-amino)-1-(2-pyrid-yl)prop-2-en-1-one-κN,O]cobalt(II) dinitrate dihydrate.

In the title compound, [Co(C(10)H(12)N(2)O)(2)(H(2)O)(2)](NO(3))(2)·2H(2)O, the Co(II) ion, located on an inversion center, is trans-coordinated by two N,O-bidentate chelating (E)-3-(dimethyl-amino)-1-(2-pyrid-yl)prop-2-en-1-one ligands and by two water mol-ecules in a slightly distorted octa-hedral geometry. Inter-molecular O-H⋯O hydrogen bonds link the cations, anions and water mol-ecules into layers parallel to the ac plane. The crystal packing also exhibits weak inter-molecular C-H⋯O hydrogen bonds.

(5,7,7,12,14,14-Hexamethyl-1,4,8,11-tetra-azacyclo-tetra-deca-4,11-diene-κN,N,N,N)(thio-cyanato-κS)nickel(II) perchlorate monohydrate.

In the title compound, [Ni(SCN)(C(16)H(32)N(4))]ClO(4)·H(2)O, the Ni(II) ion is coordinated by the four N atoms of the tetra-azacyclo-tetra-deca-4,11-diene macrocyclic ligand and by the S atom of a thio-cyanate anion. The perchlorate anion is rotationally disordered around one Cl-O bond between two orientations; the occupancies refined to 0.61 (4) and 0.39 (4). Inter-molecular O-H⋯N, N-H⋯O and N-H⋯N hydrogen bonds link two cations, two anions and two solvent water mol-ecules into a centrosymmetric cluster. The crystal packing is further stabilized by weak inter-molecular C-H⋯O hydrogen bonds.

(E)-3-Dimethyl-amino-1-(2-thien-yl)prop-2-en-1-one.

The mol-ecular skeleton of the title mol-ecule, C(9)H(11)NOS, is essentially planar: the thio-phene ring is inclined to the mean plane of the rest non-H atoms by 2.92 (3)°. The crystal packing exhibits no significantly short inter-molecular contacts.

Self-encapsulation of [MII(phen)2(H2O)2]2+ (M=Co, Zn) in one-dimensional nanochannels of [MII(H2O)6(BTC)2]4- (M=Co, Cu, Mn): a high HQ/CAT ratio catalyst for hydroxylation of phenols.

Four novel three-dimensional (3D) microporous supramolecular compounds containing nanosized channels, namely, [Co(phen)2(H2O)2]2[Co(H2O)6].2BTC.21.5H2O (1), [Co(phen)2(H2O)2]2[Cu(H2O)6].2BTC.21.5H2O (2), [Co(phen)2(H2O)2]2[Mn(H2O)6].2BTC.18H2O (3), and [Zn(phen)2(H2O)2]2[Mn(H2O)6].2BTC.22.5H2O (4), were synthesized from 1,3,5-benzenetricarboxylate (BTC), 1,10-phenanthroline (phen), and the transition-metal salt(s) by self-assembly. Single-crystal X-ray structural analysis showed that the resulting 3D microporous supramolecular frameworks consist of a two-dimensional (2D) hydrogen-bonded host framework of [MII(H2O)6(BTC)2]4- (M=Co for 1, Cu for 2, Mn for 3, 4) with rectangular-shaped cavities containing [MII(phen)2(H2O)2]2+ (M=Co for 1-3, Zn for 4) guests. The guest complex is encapsulated in the 2D hydrogen-bonded host framework by hydrogen bonding and aromatic pi-pi stacking interactions, forming the 3D hydrogen-bonded framework. The catalytic activities of 1, 2, 3, and 4 were studied using hydroxylation of phenols with 30% aqueous H2O2 as a test reaction. The compounds displayed a good phenol conversion ratio and excellent channel selectivity in the hydroxylation reaction, with a maximum hydroquinone (HQ)/catechol (CAT) ratio of 3.9.

[Infiltration of DCs and CD4+ T cells in muscle after DNA immunization].

OBJECTIVE: In order to get a better understanding of the mechanism of DNA initiating specific cytotoxicity T lymphocyte (CTL) immune response, we observed the local infiltration in injection site at different time after DNA immunization and further studied the characters of infiltration cells. METHODS: The local infiltration in injection sites was observed by HE staining at 6 h, 12 h and 1-6 days after DNA immunization. Infiltration cells were further studied with the mice dendritic cells marker CD205, T lymphocyte marker CD4 and CD8 by immunohistological methods. RESULTS: phAFP injection into mouse muscle led to a local infiltration. Infiltrates reached peak at 3-6 days after injection and appeared in several discrete clusters within the muscles. The predominant cell types found in infiltrates were macrophage-like cells and lymphocytes. Immunohistological staining revealed CD205+ and CD4+ cells. CD4+ cells could be detected in any of the injected muscles. CD205+ cells were only seen at 24 h to day 3, appearing later and disappearing earlier than CD4+ cells. However, CD8+ immunostaining positivity could not be found. CONCLUSION: Our findings indicate that the character of local infiltration after DNA immunization is different from the inflammation of injection. The recruitment of CD205+ to the injection site suggests that DCs and Th cells play an important role in DNA immunization.

[Construction and expression of anti-human AFP ScFv gene in BL-21 (DE3) E. coli].

OBJECTIVE: To construct anti-human AFP single chain fragment variable (ScFv) gene, transform it into BL-21 (DE3) E. coli for expression, and identify its bioactivity. METHODS: VH and VL genes of anti-human AFP monoclonal antibody were cloned by RT-PCR from hybridoma. The ScFv gene was spliced by sequence overlap extending (SOE) PCR, and then it was ligated into pGEM-T vector to be identified by endonuclease digestion, PCR and sequencing. ScFv gene was cloned into pET32 (a+) vector and transformed into BL-21 (DE3) E. coli. The positive clones were screened out by IPTG induction, and the ScFv antibody was purified to be identified by SDS-PAGE and competitive inhibition ELISA test. RESULTS: The VH DNA consisted of 339 bases, coming from the mouse IgG gamma chain. The VL DNA consisted of 312 bases, coming from the mouse IgG kappa chain. The VH and VL genes were spliced by 45 bases coding a (G4S)3 flexible linker. The ScFv gene consisted of 696 bases. The ScFv antibody expressed by BL-21 (DE3) fused with TrxA tag protein and formed inclusions. The relative molecular mass of TrxA-ScFv fusion protein is about 40 x 10(3) and that of ScFv is about 24 x 10(3). The ScFv antibody has excellent activity tested by competitive inhibition ELISA, the TrxA-ScFv could inhibit about 41% of the McAb to bind antigen and ScFv could inhibit about 53%. CONCLUSION: We have successfully constructed an anti-human AFP ScFv gene with 696 bases; it can express in BL-21 with high activity.

[Cloning, ligation and expression of the variable region genes of the monoclonal antibody against human HnRNPA2/B1].

OBJECTIVE: To clone the variable region genes of the monoclonal antibody (McAb) against human heterogeneous nuclear ribonucleoprotein A2/B1 (HnRNPA2/B1), ligate them to assemble single chain Fv (ScFv) gene and express in Escherichia coli. METHODS: The specificity of the anti-HnRNPA2/B1 McAb 3E8 to synthetic HnRNPA2/B1 peptide, HnRNPA2/B1 protein in lung cancer cells were examined by dot-immunobinding assay, Western blot and immunohistochemistry. The variable region genes of heavy chain (VH) and light chain (VL) were amplified from hybridoma cell by reverse transcription-polymerase chain reaction(RT-PCR), and then were linked by a linker peptide using SOE-PCR (splicing by overlap extension-PCR) to construct recombination ScFv gene. The latter was cloned into the expression vector pET28 (a+) and expressed in E coli BL21. The expressed product was identified by SDS-PAGE and competitive ELISA inhibition test. RESULTS: It was shown that the McAb combined specifically with synthetic HnRNPA2/B1 peptide and HnRNPA2/B1 protein in three lung cancer cells. The cloned VH gene and VL gene were 345 bp and 309 bp respectively and were linked successfully to obtain ScFv gene. The ScFv protein was expressed in the form of inclusion body, with molecular weight of 28,000 and immunoreactivity to HnRNPA2/B1. CONCLUSION: VH gene, VL gene and ScFv gene of anti-HnRNPA2/B1 antibody were cloned, constructed and functionally expressed in E coli. These results provide the experimental basis for elucidating the role of HnRNPA2/B1 in lung cancer.

[Construction and expression of human alpha-fetoprotein recombinant plasmid].

OBJECTIVE: To construct AFP-expressing plasmid and study the expression in eukaryotic cells. METHODS: Total RNA was isolated from fetal liver tissue. Full-length human AFP cDNA was obtained by RT-PCR amplification and then recombined into eukaryotic expression plasmid pcDNA3.1. The AFP sequence of the recombinant plasmid phAFP was determined. The AFP expressions in CHO transfected with phAFP and in muscle after injection phAFP were investigated by immunohistological methods. RESULTS: The restriction endonuclease mapping of recombinant plasmid showed 1.8 kb fragment as well as full-length human AFP cDNA, and the sequence is consistent with that from GenBank. The phAFP was successfully expressed in CHO and in muscle tissues. CONCLUSION: These results suggested that AFP can be used as a target gene of DNA vaccine in heptocellular carcinoma therapy.

[Study on the TIL and NK of IL-2 injected via pelvic retroperitoneal space in gynecological cancer patient].

OBJECTIVE: To verify the feasibility and effect of biotherapy instituted via pelvic retroperitoneal space on gynecological cancer. METHODS: Injecting IL-2 (and/or) 5-Fu through a tube installed in the pelvic retroperitoneal space. Counting the subpopulation of T cell and NK of lymph-nodes of pelvis after the drugs being by FCM. RESULTS: The numbers of CD3+, CD4+, CD8+, CD25+ and NK cells in treatment group were significantly higher than those in the control group. And the numbers of these cells in the IL-2 + 5-Fu group were significantly higher than those in the 5-Fu group. The CD25+ and NK cell numbers in the IL-2 group were significantly higher than those in the 5-Fu group (P < 0.05). CONCLUSION: The IL-2 injected via pelvic retroperitoneal space can promote the activity, development and infiltrating of T cell and NK cell in the tumor tissue.

[The effect of basic fibroblast growth factor in ovarian cancer growth and angiogenesis].

OBJECTIVE: To investigate whether basic fibroblast growth factor (bFGF) can induce the proliferation, invasion and angiogenesis of ovarian cancer or not. METHODS: Human ovarian cancer cell lines SKOV(3) 1 x 10(4)/ml were plated in 24-well dishes, bFGF at 5, 10,15 and 20 ng/ml was added and crystal violet staining was given daily for 8 days, cell numbers were counted by determining OD490. SKOV(3) cells were plated in the center of 50% extra cellular matrix gel, bFGF at 5 and 10 ng/ml was added and the migration distance of cells was measured daily. SKOV(3) 5 x 10(7)/ml were transplanted to BALB/c nude mice subcutaneous. One week later, bFGF, bFGF-MAb or 0.9% nature sodium was injected subcutaneously surrounding the tumor twice a week. Eight weeks later, the experiment ended and the volume of the tumors were measured. Intratumoral microvessel density (MVD) was measured by immunohistochemistry staining for factor VIII. RESULTS: bFGF at 0-10 ng/ml could stimulate the proliferation of SKOV(3) concentration dependently (P<0.05). On the fifth day, the cell proliferation in 10 ng/ml group was 121% above control. bFGF could stimulate the invasion of SKOV(3) concentration dependently (P<0.05). On the seventh day, the migration distance in 5 ng/ml group was 1.16 cm and 153% above control, and that in 10 ng/ml group was 1.86 cm and 245% above control. The average volume of transplanted tumors and MVD in bFGF group were 180% and 146% above control respectively those in bFGF-MAb group were 63.7% and 62.8% above control respectively. CONCLUSION: bFGF can stimulate proliferation, invasion and angiogenesis of ovarian cancer markedly; bFGF-MAb can inhibit the angiogenesis and growth of ovarian cancer.

[An experimental research in the inhibiting effect of bFGF-MAb on the growth of ovarian cancer cells and transplanted tumor].

OBJECTIVE: To investigate whether the monoclonal antibody of basic fibroblast growth factor (bFGF-MAb) inhibits the proliferation, invasion and angiogenesis of ovarian cancer. METHODS: 1. Human ovarian cancer cells SKOV3 were planted in 24-well dishes, to which was added bFGF-MAb at different concentrations, and crystal violet staining was performed daily for 8 days, then cell numbers were counted by OD490 determination. 2. SKOV3 were transplanted intraperitoneally to BALB/c nude mice. One week later, bFGF-MAb was injected intraperitoneally twice a week, and the survival time of nude mice, the number and weight of the metastatic tumors on the mensentery were observed or measured. 3. Intratumoral microvessel density(MVD) was measured by immunohistochemical staining for CD31. RESULTS: 1. bFGF-MAb inhibited the proliferation of SKOV3 in the concentration-dependent manner (P < 0.05). 2. The average survival time of nude mice in bFGF-MAb group increased by 10 days over that of control. 3. The average number and weight of metastatic tumors on mensentery in bFGF-MAb group constituted 70.6% and 69.2% of those in control group, respectively. 4. The MVD in bFGF-MAb group accounted for 62.8% of the MVD in control group. CONCLUSION: bFGF-MAb can inhibit the proliferation and angiogenesis of ovarian cancer markedly, so it promises to have application in the biological treatment of ovarian cancer.