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.

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.

[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.

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

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

The aim of this study is to improve liposome encapsulation efficiency of water soluble drug ATP-2Na with hydrophobic ion pairing method, and evaluate its effect on tissues energy state in myocardial ischemia mice. Ion pair complex of ATP-2Na with HTAB was prepared first; then the liposomes were prepared by ethanol injection method. The size and zeta potential of ATP-2Na liposome were investigated. Its effect on tissues energy state in myocardium ischemia mice was evaluated by detecting ATP-2Na concentration in tissues and blood after injection in comparison to ATP-2Na solution. The diameters and zeta potential of ATP-2Na liposomes were (144.0 +/- 2.7) nm and (+16.2 +/- 1.6) mV, respectively. The encapsulation efficiency was (85.02 +/- 2.31) %. The in vitro drug release pattern from liposomes matches with Weibull equation. Compared with ATP-2Na solution, ATP-2Na liposome increased the ATP concentration of blood in myocardial ischemic mice very significantly; compared with blank, ATP-2Na liposome increased ATP content of myocardium and liver in myocaridal ischemic mice significantly, but ATP-2Na solution didn't show this effect. ATP-2Na liposome might have an advantage in improving tissue energy state on myocaridal ischemic animals.
Adenosine Triphosphate ; administration & dosage ; blood ; metabolism ; Animals ; Cetrimonium Compounds ; chemistry ; Drug Carriers ; Liposomes ; chemistry ; Liver ; metabolism ; Male ; Mice ; Myocardial Ischemia ; blood ; metabolism ; Myocardium ; metabolism ; Particle Size ; Random Allocation ; Surface-Active Agents ; chemistry

BACKGROUND:There is a great difference of grade size of macrobead in various joint diseases; therefore, it can be used to determine state of joint diseases initially.OBJECTTVE : To explore the physical properties of synovial fluid nano-particles and their correlations with the occurrence of knee osteoarthritis (KOA).DESIGN: Controlled experimental study based on synovial fluid samples.PARTICIPANTS: A total of 99 synovial fluid samples were collected from normal subjects and KOA patients with various KOA severities. Among them, 41 were normal synovial fluids, 58 were KOA.METHODS: Synovial fluid samples from individuals with and without KOA were obtained. Using the technology of quasi-elastic laser scattering, nano-particle size and its distribution were estimated, and the dynamic/static light scattering spectrometric analyzer allowed the measurement of particles Zeta potentials. A correlation analysis between the particle size, Zeta potentials and the onset of KOA was attempted.MAIN OUTCOME MEASURES:① Grade size and distribution of microsome in synovial fluid;② Zeta potentials and distribution of microsome in synovial fluid; ③ grade size and clinical correlation of microsome in synovial fluid.RESULTS: ① The mean nano-particle diameter in the synovial fluid of KOA patients were significantly greater than those of normal joints [(297±84), (63±23) nm, P ＜ 0.001]. The distribution curve of KOA synovial fluid nano-particle size was normal knee and (-15.84 ±3.34) mV of KOA patients, and there was a significant difference (P ＜ 0.001). This suggestedthat the Zeta potentials in the synovial fluid of KOA patients were significantly greater than those of normal joints. ③ The average particle size and Zeta potential of synovial fluid strongly correlated with the integrity of the joint of KOA (rp =0.797 2,0.631 9, P＜ 0.01).CONCLUSION: The nano-particle size and Zeta potential of synovial fluid are significantly correlated with the development of KOA, and this can reflect the severity of KOA.

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.