Objective: To study the effects of capsaicin applied locally to infraorbital nerve on primary afferent nerve inadult rats. Methods:Capsaicin solution (2% ) was directly applied onto one lateral intraorbital nerve, and the contralateralnerve was used as self-control. Both experimental and control infraorbita1 nerves were observed under electron tnicroscope on3, 7, 14, 21, 28, 35 or 42 d after local administration. Substance P(SP)--like immunoreactive positive granules in both lateralnucleus caudalis spinalis nervi trigemini (CNV) were examined by immunohistochemical method. Results: Degenerated un-myelinated fibers in capsaicin-treated nerves were found under electron microscope, and no myelinated fibers changes werefound. The gray scale value of SP-like immunoreactive positive granules in the experimental side was markedly lower thanthat in the control side (P＜0. 01), while no significant difference was observed between different time groups. Conclusion:Local administration of capsaicin can produce selective destruction of unmyelinated fibers in primary afferent nerves, and canreduce the levels of SP in regions of CNV. The results suggest that local administration of capsaicin has potential therapeuticvalue for trigeminal neuralgia.

Objective To explore the differences and similarities of the cervical lesions and mechanism between Asian variant E6 T178G and European variant E6 T350G, A442C and other variants. Methods We selected 300 clinic or hospitalized patients in our hospital during the period of May 2011 to October 2012. Cervical exfoliated cells were harvested by Thinprep cytologic test (TCT). A PCR sequencing assay was performed to detect HPV16 E2, E6 and E7 gene variants. One year later, the test was repeated. The patients with persistent infection underwent cervical biopsy by colposcopy for pathological examination. SP immunohistochemical method was applied to detect E7 protein expression level in all the patients. Results After one year, of 292 patients who were successfully sequenced, 259 were chronic cervicitis, 32 were cervical intraepithelial neoplasia grade I (CINI), and one was cervical intraepithelial neoplasia grade II (CINII). E7 protein expressed in each variant. But the expression of E7 protein in patients with different variant infection had no significant difference from each other. Conclusions E7 protein may be play a role in the early stages of HPV16?induced cervical lesions. But E7 protein may not be a reference index of the different carcinogenic mechanism between different HPV16 variants.

Human acute premyeloid leukemia cell cDNA expression library was constructed to screen acute premyeloid leukemia tumor antigen. Total RNA and purified mRNA were extracted from human premyeloid cell line NB4. First and second strands of cDNA were synthesized by reverse transcription. After blunting, the cDNA fragments were ligated with EcoR I adapters. Then the cDNAs were digested with Xho I, and less than 400 bp cDNA fragment was removed by Sephacryl-S400 spin column, the remaining were ligated with lambdaZAP vector. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli XL1-Blue-MRF' for titration. The recombinants were examined by color selection. In order to evaluate the size of cDNA inserts and the diversity of library, the pBK-CMV phagemid was excised from the ZAP express vector by using ExAssist helper phage with XLOLR strain, and then the pBK-CMV phagemid was digested by Xho I and EcoR I. The results showed that the NB4 cell line cDNA library consisting of 1.65 x 10(6) recombinant bacteriophages was constructed with the recombinant ratio of 99.6%. The average length of the recombinant exogenous inserts was about 1.7 kb. It was concluded that the constructed cDNA library are deserved to screen target clones.
Bacteriophages/genetics ; DNA, Complementary/*biosynthesis ; *DNA, Neoplasm/biosynthesis ; DNA, Recombinant/biosynthesis ; *Gene Library ; Genetic Vectors ; Leukemia, Promyelocytic, Acute/*genetics ; Leukemia, Promyelocytic, Acute/metabolism ; Leukemia, Promyelocytic, Acute/pathology ; RNA-Directed DNA Polymerase/metabolism ; Transcription, Genetic/genetics

OBJECTIVETo study the conjugation reaction characteristics of caffeic acid micromolecule cistanoside F and bovine serum albumin.

METHODThe interaction between bovine serum albumin (BSA) and cistanoside F that was separated from Callicarpa plant for the first time and abbreviated CF was detected by fluorescence (FS), UV-vis absorbance and circular dichroism (CD) under simulative physiological conditions.

RESULTCF-BSA's static apparent binding constant (K(a)), number of binding sites (n), efficiency of energy transfer (E), spatial distance (r), thermodynamic parameters deltaG, deltaH and deltaS and changes in alpha-helical structure content in BSA before and after CF's effect were calculated to define the binding site of CF in BSA and analyze the impact of several common metal ions on the interaction of CF and BSA.

CONCLUSIONGround state compounds formed by CF and BSA could cause intrinsic fluorescence quenching. Their binding constant K(a) of cistanoside F with BSA was 4.36 x 10(4) L x mol at 25 degrees C, the number of binding site n was 1, and the spatial distance r was 3.09 nm. The results indicated that the hydrogen bond played a major role in cistanoside F-BSA association. The displacement experiments confirmed that cistanoside F can bind to site I of BSA. In addition, the binding constant of cistanoside F with BSA was enhanced after the addition of some common metal ions Mg2+, Fe3+, Cu2+ and Zn2+. The intrinsic fluorescence of BSA was quenched by cistanoside F via forming cistanoside F-BSA complex and non-radiation energy transfer. CD spectra showed that the binding of cistanoside F with BSA induced conformational changes in BSA.

Animals ; Caffeic Acids ; chemistry ; Catechols ; chemistry ; Cattle ; Circular Dichroism ; Glycosides ; chemistry ; Serum Albumin, Bovine ; chemistry ; Spectrometry, Fluorescence ; Spectrophotometry, Ultraviolet ; Thermodynamics

Human acute premyeloid leukemia cell cDNA expression library was constructed to screen acute premyeloid leukemia tumor antigen. Total RNA and purified mRNA were extracted from human premyeloid cell line NB4. First and second strands of cDNA were synthesized by reverse transcription. After blunting, the cDNA fragments were ligated with EcoR I adapters. Then the cDNAs were digested with Xho I, and less than 400 bp cDNA fragment was removed by Sephacryl-S400 spin column, the remaining were ligated with lambdaZAP vector. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli XL1-Blue-MRF' for titration. The recombinants were examined by color selection. In order to evaluate the size of cDNA inserts and the diversity of library, the pBK-CMV phagemid was excised from the ZAP express vector by using ExAssist helper phage with XLOLR strain, and then the pBK-CMV phagemid was digested by Xho I and EcoR I. The results showed that the NB4 cell line cDNA library consisting of 1.65 x 10(6) recombinant bacteriophages was constructed with the recombinant ratio of 99.6%. The average length of the recombinant exogenous inserts was about 1.7 kb. It was concluded that the constructed cDNA library are deserved to screen target clones.
Bacteriophages ; genetics ; DNA, Complementary ; biosynthesis ; DNA, Neoplasm ; biosynthesis ; DNA, Recombinant ; biosynthesis ; Gene Library ; Genetic Vectors ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; metabolism ; pathology ; RNA-Directed DNA Polymerase ; metabolism ; Transcription, Genetic ; genetics