[Summary] The genomic DNA was extracted from peripheral blood leukocyte of the patient with thyroid hormone resistance syndrome and 14 members of his family.The exons 1-10 of thyroid hormone receptor β (TRβ) gene were amplified by PCR.The products of PCR were sequenced directly to detect the gene mutation.The results showed that 3 members of this family were confirmed to have the C→T transition mutation at nucleotide 1 303 site within exon 10 of TRβ gene,and the missense mutation results in the substitution of histidine to tyrosine (H435Y).The heterozygous mutation may lead to the occurrence of thyroid hormone resistance syndrome.

Objective To compile "care operation and integration of nurse-patient communication program"and discuss its clinical application effect in older to improve the nursing care operations, communication and ability to communicate with patients and building a harmonious relationship between nurses and patients to improve quality of care. Methods We investigated and studied the problems existed in communicating with patients during care operations and put forward the concept of operation and integration of communication, compiled "nursing operation and integration of nurse-patient communication procedures" and put it into clinical application. Results Satisfaction degree surveys after application showed that patients' satisfaction degree increased and communication skills of nursing students increased. Conclusions Application of "nursing and the nurse-patient communication integration process" improved patients' satisfaction degree and make the nurse-patient relationship more harmonious.

Objective To investigate the expression and the effects of X-chromosome linked inhibi- tor of apoptosis (XIAP) associated factor 1 (XAF1) on apoptosis and cell proliferation in SMMC7721 hepatocellur carcinoma (HCC) cell line.Methods The expression of XAF1 mRNA and protein in hu- man SMMC7721 cell line were detected by semi-quantitative,RT-PCR and Western blot.Plasmid con- structs expressing sense and antisense XAF1 were generated and transfected into SMMC7721 cell line to establish stable transfectants.Cell proliferation and apoptosis were detected by MTT,flow cytometry and TUNEL.Results XAF1 mRNA and protein were detectable in SMMC7721 cell line but lower than that in normal liver cell.Stable expression of XAF1 significantly inhibited cell proliferation and increased spontaneous apoptosis in SMMC7721 cell (P＜0.05).Over-expression of XAF1 in stable transfactants increased the sensitivity of SMMC7721 cell to chemotherapeutic drugs such as 5-fluorouracial and hydroxycamptothecin.Conclusions Over-expression of XAF1 induces apoptosis and inhibits SMMC7721 cell growth.XAF1 may be a promising candidate for HCC gene therapy.

OBJECTIVETo explore the ultrastructural changes of hepatocyte fibrogenesis in cholelithiasis in biliary tract.

METHODSl0 liver biopsies were taken from the patients suffered from gallstone and choledocholithiasis during surgical treatment and the ultrastructural changes were observed under electromicroscope.

RESULTSThere were plentiful collagenous microfibrils (CMFs) grown within some hepatocytes. These CMFs distributed locally or diffusely in cytoplasm even extended into nucleus. In 7 cases numerous megamitochondrias appeared in several hepatocytes, the inclusions mimicking fibrils could be frequently seen and grew beyond the envelope. Furthermore, typical CMFs could be seen in the large microbodies, and several vesicular or cystic structures similar as fibroblast were presented in marginal areas of the hepatocytes.

CONCLUSIONSWe deduce that the fibrosed hepatocytes may be remained and take part in the hyperplasia of hepatic fibrous tissue.

Adult ; Cholelithiasis ; pathology ; ultrastructure ; Female ; Hepatocytes ; pathology ; ultrastructure ; Humans ; Liver Cirrhosis ; pathology ; Male ; Middle Aged

To establish single- and double-transfected transgenic cells stably expressing hMATE1, hMATE1 cDNA was cloned by RT-PCR from human cryopreserved kidney tissue, and subcloned into pcDNA3.1(+) plasmid by virtue of both HindIII and Kpn I restriction enzyme sites. Subsequently, the recombined pcDNA3.1(+)- hMATE1 plasmid was transfected into MDCK, MDCK-hOCT1 or MDCK-hOCT2 cells using Lipofectamine 2000 Reagent. After a 14-day-cultivation with hygromycin B at the concentration of 400 µg · mL(-1), all clones were screened with DAPI and MPP+ as substrates to identify the best candidate. The mRNA content of hMATE1, the cellular accumulation of metformin with or without cimetidine as inhibitor, or transportation of cimetidine was further valuated. The results showed that all of the three cell models over expressed hMATE1 mRNA. The cellular accumulation of metformin in MDCK-hMATE1 was 17.6 folds of the control cell, which was significantly inhibited by 100 µmol · L(-1) cimetidine. The transcellular transport parameter net efflux ratios of cimetidine across MDCK-hOCT1/hMATE1 and MDCK-hOCT2/hMATE1 monolayer were 17.5 and 3.65, respectively. In conclusion, cell models with good hMATE1 function have been established successfully, which can be applied to study the drug transport or drug-drug interaction involving hMATE1 alone or together with hOCT1/2 in vitro.
Animals ; Biological Transport ; Cimetidine ; pharmacology ; DNA, Complementary ; Dogs ; Drug Interactions ; Humans ; Madin Darby Canine Kidney Cells ; Metformin ; pharmacology ; Organic Cation Transport Proteins ; genetics ; metabolism ; Transfection

The biochemical mechanism of degrading keratins by S.fradiae var S-221 was primarily studied.The compounds (Na_ 2 SO_ 4 , Na_ 2 SO_ 3 and sulfdryl acohol), which respecitively enhance specific activity of keratinase, activate keratinase intensively and mainly act on the disulfide bonds reductase in the keratinase, Na_ 2 SO_ 3 activates intensively both disulfide bonds reductase and polypeptide hydrolytase at 0.01 mol/L, whereas Na_ 2 S_ 2 O_ 3 , which acts on the disulfide bonds reductase, inhibits keratinase.On the condition that substrate, keratins exists, S.fradiae var S-221 is induced to produce exo-keratinase, which is a multiproteinase, containing disulfide bonds reductase, which is a key enzyme degrading keratins, then, with polypeptidic, hydrolytase, graduately hydrolyzates denatured keratins into polypeptides, oligopeptides and free amino acids, so that keratins have been decomposed completely.Sulfur in the keratins was transferred into sulfhydryl compounds, H_ 2 S and sulfates in the course of keratinolysine.

Objective To develop a nylon membrane chip for rapid and systematic detection of the diabetes-associated 45 mutant loci in mitochondrial DNA(mtDNA).Methods The mutant-and wild-type probes were designed for detection of 45 mutant loci in mtDNA with Primer Premier 5.0 and NCBI BLAST softwares and the 90 probes with 8 poly T were immobilized on the Hybond N~+ nylon membranes which were treated with 5?SSC Buffer by UV-crosslinking;Then asymmetric PCR was employed to obtain the target single strand DNA(ssDNA).The PCR products were labeled with biotin after purification.NBT/BCIP was used as substrate that yields a very intense purple signal followed by AP-avidin,and the signals were observed in 24 samples with known sequences to evaluate the chips,each sample was repeatedly measured three times.Results The specific target fragments of 45 loci can be amplified under the same condition with nine sets of primers.The annealing temperatures of the wild-type [(59.01?1.42)℃] and mutant-type [(59.34?1.29)℃ ] probes are so close(t=1.046,P =0.301)that hybridization can be performed at the same temperature.The spots on the membrane chip are distinct,regular and well-distributed.The results of positive-and negative-control are perfect.The signals of negative probes and the background are similar.The results of chip were nearly concordant with that of DNA sequences(?~2=113.132,Kappa value =0.888,P = 0.000)except for T16189C mutant.Conclusions We have successfully developed a nylon membrane chip for rapid and systematic detection of the diabetes-associated 44 mutant loci in mtDNA.It could be used for screening for diabetic patients and high-risk people.

To construct a secretory-expression vector of antimicrobial peptide Bactenecin 7(Bac7),and identify the secretory-expression product in L.lactis MG1363 and its bioactivity.The splicing primers of regulation elements and Bac7 gene,which designed according to codon usage preferences of L.lactis MG1363,were chemically synthesized,and the overlap-extension PCR method was used to splice the full length of Bac7 gene.Then the Bac7 gene was linked to expression vector pMG36e to construct pMG36e/Bac7 vector,and pMG36e/Bac7 was transformed into L.lactis MG1363 by electrophoration.RT-PCR and Western blot assays were applied to investigate the expression of the Bac7 gene in L.lactis,and bioactivity of Bac7 in culture supernatant of L.lactis was tested with plate-diffusion method.The results showed that the Bac7 gene and its regulation elements was amplified and cloned in the vector pMG36e successfully,The secretory-expressed Bac7 in L.lactis MG1363 harboring pMG36e/Bac7 was identified by Western blot,and it had high bacteriostatic activity against E.coli.These results indicate that the recombinant L.lactis MG1363 could express bioactive Bac7,which lays a foundation for further study of oral administration of a Bac7-secreting L.lactis to treat intestinal bacteria infection.

ObjectiveTo evaluate the treatment efifcacy of the treatment promotion of standardized management for chil-dren with asthma.MethodsMedical records of 150 children with asthma were reviewed and divided into management group or control group according to whether standardized management was accepted. Comprehensive asthma education for asthma pa-tients and their parents including asthma associated basic knowledge education, health education as well as follow-ups at deifned intervals was conducted in 78 cases. In the meantime, standardized asthma therapies were performed. Control group involved 72 cases who did not receive asthma education managements and only accepted regular clinical therapies. After 1-year observational follow-up, , clinical efifcacy of children with asthma, changes of knowledge-attitude-practice of parents, and compliance of med-ication were compared between the two groups.ResultsAfter promotion of standardized managements treatment, asthma con-trol rates in the management group were signiifcantly higher than that of the control group(χ2=54.68,P<0.01); In addition, the rate of asthma attacks, emergency visits as well as hospitalizations were obviously reduced in the management group than control group (both withP<0.01). Knowledge associated with asthma, therapy and management executions as well as knowledge-atti-tude-practice of parents also demonstrated apparent elevations in the management group (P<0.01); At the same time, management group has illustrated superior medication compliance over the control group (χ2=66.27,P<0.01).ConclusionPromotion of standardized treatment management among children with asthma can help to achieve effective control by raising levels of knowl-edge-attitude-practice of the parents as well as the patient’s compliance to the treatment.

[ABSTRACT]AIM:Toexploretheeffectoftheelasticmodulusandsizesofliquidcrystal(LC)phasesonosteo-genic differentiation based on OPC/PU composite substrate by mimicking the microenvironment in rat bone mesenchymal stem cells (rBMSCs).METHODS: A series of composite substrates with different elastic modulus were constructed via modulation of LC content in the composites .The surface phase structure was observed by polarized microscopy , and the mechanical property was measured by a universal material testing machine .Furthermore, the laser confocal microscope was employed to observe the spreading , polarization and the cytoskeleton arrangement of the rBMSCs .The proliferation of rBM-SCs was evaluated by CCK-8 assay.The specific mRNA expression of osteogenic differentiation such as collagen Ⅰ, and osteopontin on the composite membranes was detected by real-time PCR.RESULTS:The size and number of LC phase in-creased and the elastic modulus of the composite substrates decreased with the increase of the LC content .The rBMSCs ex-hibited better characteristics of initial adhesion , spreading and proliferation on the OPC 10-PU and OPC30-PU in the early and medium culturing .The rBMSCs displayed higher expression of collagen Ⅰ and osteopontin on the OPC10-PU in the early and medium osteogenic induction , while the high expression of these osteogenic genes occured on the OPC 30-PU and OPC50-PU in later osteogenic induction .The emphasis of genetic expression was switched from collagen Ⅰin the early and medium osteogenic induction to osteopontin in the later stage .CONCLUSION:When the content of LC remained low in the composite substrates , rBMSCs mainly responded to the mechanical stimuli induced by substrate stiffness and exhibited distinguished cellular behaviors;with the increase in the LC content , rBMSCs had strong interactions with LC by sensing the viscoelasticity of LC , probably resulted from the contribution of both substrate stiffness and the viscoelasticity of LC phase .