Adult ; Ammonia ; poisoning ; Female ; Humans ; Microscopy, Electron ; Poisoning ; complications ; Respiratory Function Tests ; Respiratory System ; drug effects ; pathology ; ultrastructure ; Time Factors

Humans ; Stomatitis, Aphthous ; diagnosis ; etiology ; therapy

AIM: To investigate the effects of Tangmoning (TMN) Granule on nerve conduction velocity, red blood cell sorbitol (RBC S) level in diabetic rats and probe the mechanism of prevention and treatment of diabetic peripheral neuropathy. METHODS: The model of diabetic rat was induced by strepotozotocin. The animals were randomly divided into six groups:TMN minimal dose group, TMN mild moderate dose group, TMN maximal dose group, methycobal group, model group and normal control group. All TMN groups were treated with TMN Granule through tube feeding (2.95 g/kg、5.90g/kg、11.80g/kg). Methycobal group was treated with methycobal tablets through tube feeding (0.14mg/kg). The duration of treatment was 3 weeks. The caudal nerve conduction velocity, RBC S content were investigated before and after treatment. RESULTS: It showed that TMN Granule could increase the caudal nerve conduction velocity significantly in the maximal dose group and the mild moderate dose gorup ( P

Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.

To clone, express and primarily use human autoantigen Sm D1 in methylotrophic yeast Pichia Pastoris. The gene Sm D1 was cloned by PCR.The PCR product was inserted into the vector pPIC9k. The recombinant plasmid pPIC9k- Sm D1 was transformed into yeast SMD1168 by electroporation. The positive clones were screened in MD plates. The high copy number transformants were rapidly selected by using G418 and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and im-munodot. The PCR product was showed about 360 bp in size which was in accordance with predicted. The pPIC9k-Sm D1 showed the same seqencing result with GenBank’s report and restriction enzyme analysis confirmed our prediction. The pPIC9k-Sm D1 positive clone produced an about 16 kD protein which had natural immunogenicity of human autoantigen Sm D1 by SDS-PAGE and immunodot. The sensitivity and specificity of immunodot were 96% and 100%, respectively. The agreement between immunodot and im-munoblot was 98%. Successfully cloning and high-level expression of human autoantigen Sm D1 in methy-lotrophic yeast Pichia pastoris laid a foundation for further research work.

Objective To clone and express HCMV UL123 gene exon 2,3(ie1-exon2,3) in bacterial two-hybrid bait plasmid,and identify its self-activation property.Methods HCMV ie1-exon2,3 carried on pTWIN1/ie1 recombinant was amplified by PCR and cloned into the pBT plasmid,which was transformed into E.coli XL1-Blue MRF' Kan host strain.The positive recombinant was identified by PCR,restriction enzyme digestions and sequencing analysis.The verified plasmid was transformed into bacterial two-hybrid system reporter strain.The soluble fusion protein was analyzed by SDS-PAGE and Western blot.The self activation effect of the recombinant was then tested.Results Bacterial two-hybrid pBT/ie1-exon2,3 bait plasmid was successfully constructed.The corresponding soluble fusion protein rIE1-N_(85)/?C1 was expressed in bacterial two-hybrid system reporter strain,and didn't show self-activation property.Conclusion Bacterial two-hybrid pBT/ie1-exon2,3 bait plasmid without self-activation property was successfully constructed,and it can be used to screen the library of fetal brains.

AIM: To explore the clinical effects of high frequency electrical capsulotomy in maturation period cataract surgery.
●METHODS: A total of 68 cases of maturation period cataract were selected and underwent the surgery of continuous circular capsulorhexis using the high frequency electrical capsulotomy.
●RESULTS: The success rate was 91% in 68 cases with the high frequency electrical capsulotomy.
● CONCLUSION: The high frequency electrical capsulotomy in maturation period cataract surgery has significant advantages and brilliant clinical values.

OBJECTIVETo investigate the protective effects of Jiedu Tongluo injection on cerebral edema induced by focal lesion of cerebral ischemia/reperfusion, the hydrous content of brain and the expressions of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin and MMP-9 in rats.

METHODThe model of brain middle cerebral artery ischemia/reperfusion was established by the thread approach. After 24 hours of reperfusion, cerebral edema formation was determined by the hydrous content of brain. The permeability of blood brain barrier was evaluated based on the leakage of Evans blue. Enzyme-linked immunoadsordent assay (ELISA)was used to examine the expression of ICAM-1, VCAM-1, E-selectin. The expression of MMP-9 was measured by immunohistochemistry.

RESULTJDTL, in the dose of 2 mL x kg(-1) and 4 mL x kg(-1), relieved cerebral edema (P < 0.05, P < 0.01), reduced the expressions of ICAM-1, VCAM-land E-selectin and decreased MMP-9 activity (P < 0. 05, P < 0.01) in model rats.

CONCLUSIONJiedu Tongluo injection has a protective effect on rat brain from cerebral edema induced by the injury of focal cerebral ischemia/reperfusion. The mechanism is related to that Jiedu Tongluo injection can reduce the expressions of ICAM-1, VCAM-1 and E-selectin and inhibit of MMP-9 activation in rat brain.

Animals ; Blood-Brain Barrier ; drug effects ; metabolism ; Brain Edema ; etiology ; metabolism ; prevention & control ; Brain Ischemia ; complications ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; E-Selectin ; metabolism ; Evans Blue ; metabolism ; Gene Expression Regulation, Enzymologic ; drug effects ; Injections ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Permeability ; drug effects ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; Vascular Cell Adhesion Molecule-1 ; metabolism

Objective To compare the relative efficacy and quality of extraction of human fecal DNA using four methods.Methods Real-time PCR were utilized for analysis both quantification and quality of the fecal targeted bacteria(including gut all eubaeterium,Bacteriodes-PrevoteUa group,Bifidobacterium spp Enterobacteriaceae and Enterococcus spp)by using 16s rRNA gene-targeted genus or group-specific primer sets.Results The negative rat of PCR product from method 3(phenol-chloroform plus bead-beating) was about 40%(4/10)by using universal primers,the PCR inhibition disappeared after fecal DNA purified with column.The total fecal 16s rRNA gene copy numbers(per gram of wet weight of feces)as well as the numbers of Bacteriodes-Prevotella group from method 1(QIAamp~DNA stool mini kit)and 4(QIAamp~ DNA stool mini kit combined with bead-beating)was higher significantly than that from method 2(FastDNA ~Kit,Biol01)and 3(P

Objective To investigate the effects of human S100A6 on β-catenin in human osteosarcoma cell lines MG63 and U2OS. Methods Cell lines MG63 and U2OS were infected by recombinant adenoviruses carrying human S100A6 and its siRNA gene, AdS100A6 and AdSiS100A6 respectively, to up-regulate and down-regulate the ex-pression of S100A6. Then RT-PCR, Western blot and immunocytochemistry were used to detect mRNA and protein (level and/or distribution) of β-catenin. Results In both cell lines, with up-regulated S100A6, expression of β-catenin mRNA and protein increased(P <0. 05) and β-catenin protein increase was more obvious in nuclear than in cytoplasma; while down-regulating S100A6, both the mRNA and protein level of β-catenin decreased (P<0. 05) ; β-catenin protein decrease was more obvious in nuclear than in cytoplasma, too. Conclusion In-creasing Wnt/β-catenin signaling activity may be a mechanism that S100A6 involves in tumor development.