Adeno-associated virus-mediated BMP-7 and SOX9 in vitro co-transfection of human degenerative intervertebral disc cells.

Bone morphogenetic protein-7 (BMP-7) and SOX9 are important transcription factors in chondrogenesis. In this study, we examined the biological function of the adeno-associated virus (AAV)-mediated BMP-7 and SOX9 double gene in vitro co-transfection of nucleus pulposus cells of human degenerative intervertebral disc. Human intervertebral disc nucleus pulposus cells were cultured in vitro and subcultured for 5 generations. Using rAAV-BMP-7 and rAAV-SOX9 AAV2-type AAV viruses, the cells were divided into 4 groups: blank normal, BMP-7 transfection, SOX9 transfection, and BMP-7 and SOX9 co-transfection. After 48 h, expression of type II collagen and its mRNA in nucleus pulposus cells was determined. The expression of type II collagen in BMP-7 transfection, SOX9 transfection, and co-transfection groups was up-regulated to varying degrees compared to the blank control group. The type II collagen mRNA level expression in the co-transfection group was significantly higher than in other groups (P < 0.05). AAV-mediated BMP-7 and SOX9 in vitro co-transfection can promote the synthesis of type II collagen in nucleus pulposus cells in the human degenerative intervertebral disc. Double-gene therapy has a synergistic effect in the treatment of intervertebral disc degeneration.

Lipopolysaccharide and ß-1,3-glucan binding protein in the hard clam (Meretrix meretrix): molecular characterization and expression analysis.

Pattern recognition molecules play an important role in innate immunity by recognizing conserved molecular patterns that are present on the surface of invading microorganisms. In this study, a lipopolysaccharide and ß-1,3-glucan binding protein (LGBP) gene was cloned from the hard clam Meretrix meretrix (designated as Mm-LGBP) by the expressed sequence tags and rapid amplification of cDNA ends method. The cDNA was 1827 bp in length, consisting of a 71-bp 5'-terminal untranslated region, a 62-bp 3'UTR, and a 1734-bp open reading frame encoding a 577-amino acid polypeptide with an estimated molecular mass of 60.7 kDa and a theoretical isoelectric point of 5.56. Characteristic potential polysaccharide binding, cell adhesion, and glucanase motifs were identified in the Mm-LGBP, indicating that Mm-LGBP should be a new member of the LGBP family. Quantitative real-time polymerase chain reaction was developed to detect the mRNA expression level of Mm-LGBP in 6 different tissues. Higher-level mRNA expression of Mm-LGBP was detected in the gill and digestive gland tissues. The upregulation of Mm-LGBP mRNA after Vibrio anguillarum challenge showed that Mm-LGBP play a pivotal role in antibacterial immunity.

Determination of aspartame by ion chromatography with electrochemical integrated amperometric detection.

In this paper, the separation and determination of the sweetener aspartame by ion chromatography coupled with electrochemical amperometric detection is reported. Sodium saccharin, acesulfame-K and aspartame were separated using 27.5 mmol/l NaOH isocratic elution on a Dionex IonPac AS4A-SC separation column. Aspartame can be determined by integrated amperometric detection without interference from the other two sweeteners. The method can be applied to the determination of aspartame in powered tabletop, fruit juice and carbonated beverage samples, and the results obtained by integrated amperometry were in agreement with those obtained using a UV detection method. A method for determining analytes with an NH2 group by ion chromatography with integrated amperometry was developed.

Neural representation of sound duration in the inferior colliculus of the mouse.

In this study we examined the neuronal responses of single units to different sound durations in the inferior colliculus (IC) of the mouse. One hundred and one recorded units were classified into onset (58%), sustained (9%) on-sustained (22%), pauser (9%) and chopper (2%) response patterns. Thirty-four percent of the recorded units showing stronger responses to long stimulus durations were defined as long-duration-selective neurons. Twenty-five percent of the units preferred a narrow range of sound durations and were classified as band-pass neurons. Ten percent of the units responded preferentially to short stimulus durations and thus displayed short-duration selectivity. Twelve percent of the units that responded with nearly constant spike counts to stimuli of varying duration were classified as all-pass neurons. In contrast to the result of no short-duration-selective neurons found in chinchilla IC, we observed that some of the onset units in the IC of the mouse displayed a short duration preference. The best duration range of the duration-selective neurons in the present study corresponds to the duration range of mouse calls. We suggest that an inhibitory mechanism contributes to the duration selectivity observed in the present study.

[The mechanism of renal hyperperfusion induced by hyperglycemia].

Using whole body clearance technique as well as isolated perfused rat kidney (IPRK). We studied the effect of hyperglycemia on renal hemodynamics and the possible mechanism involved. Hyperglycemia increased renal plasma flow (RPF) and glomerular filtration rate (GFR). In nonfiltering isolated perfused rat kidney or after adding furosemide to the perfusate, both techniques were believed to abolish the tubuloglomerular feedback, the renal hemodynamic changes induced by hyperglycemia disappeared, but adding sodium nitroprusside increased RPF and decreased renal vascular resistance (RVR). D-alpha-methyl-glucoside, a glucose derivative, increased RPF and GFR in IPRK, but had no influence on renal hemodynamics in whole body clearance study. Hemoglobin, which could block the action of endothelium--derived relaxing factor (EDRF), did not change the effects of hyperglycemia on renal hemodynamics in IPRK. These results indicated that hyperglycemia could directly induce hyperperfusion and hyperfiltration, which is mainly mediated by suppression of tubuloglomerular feedback. EDRF do not play the major role in the changes of hemodynamics induced by hyperglycemia.

Studies on the structure of HBV DNA.

The structure of HBV adr NC-1 DNA is analyzed and compared with another five strains of HBV DNAs. Some of the prokaryotic promoter-like sequences, palindrom sequences and ATAA are found. An enhancer core sequence and some other characteristics are also shown. In considering the frame and its regulatory sequence as a transcriptional unit some of the possible new frames are discussed.

Determination of the complete nucleotide sequence of HBV adr NC-1 DNA.

On the basis of sequencing the large DNA-fragments which have been inserted into M13mp8, we design a simple strategy to determine the complete nucleotide sequence of HBV adr NC-1 DNA with chain termination method. The whole genome is 3195 nucleotides long. Five reading frames are observed. The gene location and organization are shown.

[Experimental study on Jurkat cell apoptosis induced by Boswellia carterii Birdw extractive].

OBJECTIVE: To study the influence of Boswellia carterii Birdw(BCB) extractive of different concentrations on human Jurkat cell apoptosis at different time points. METHODS: Agarose gel electrophoresis, transmission electron microscope(TEM) and flow cytometry(FCM) were used for observing DNA ladders, morphology of Jurkat cells and cell cycle, respectively. RESULTS: BCB induced apoptosis of Jurkat cells and typical DNA ladders; TEM demonstrated the presence of apoptosis Jurkat cells, condensation of cytoplasm, numerous vaculoes in cytoplasm, compaction of the nuclear chromatin and formation of apoptosis dody. The sub-G1 peak was detected by cell cycle analysis. It revealed that G1 phage cells increased and S phage cells decreased. CONCLUSION: The data indicate that BCB extractive induces time- and concentration-dependent apoptosis in Jurkat cell line.

[Study on the differentiation and apoptosis of HL-60 cell line induced by Puerarin].

OBJECTIVE: To investigate the differentiation of HL-60 cells induced by different doses of Purerarin(PR) comparing with all-trans retinoic acid(ATRA) and to see if PR can induce apoptosis of HL-60 cells. METHODS: Cell differentiation was analyzed by NBT reduction, the ratio of NBT/MTT and CD11b, apoptosis by morphology, DNA electrophoresis, and flow cytometry(FCM). RESULTS: 80 micrograms.ml-1, 160 micrograms.ml-1, 320 micrograms.ml-1 PR could induce differentiation of HL-60 cells, no significant difference was observed between the cells treated with 1 mumol.L-1 ATRA and 320 micrograms.ml-1 PR. Treated with 320 micrograms.ml-1, 640 micrograms.ml-1 PR, HL-60 cells exhibited a morphological characteristic of apoptosis and typical DNA ladder on gel electrophoresis. FCM analysis showed that PR could interfere with cell cycle in HL-60 cells, with a increased ratio of sub-G1 in HL-60 cells. CONCLUSION: PR exerts effect on differentiation and induces apoptosis in HL-60 cells.

[The relationship between clinical prognosis and classification of the isoforms of fuse gene in acute promyelocytic leukemia].

To check the anti-apoptosis gene bcl-2 protein/mRNA expression in 28 acute promyelocytic leukemia (APL), by immunohistochemistry and in situ hybridization, and to analyse the relationship among the bcl-2 gene expressions, clinical prognosis and classification of the isoforms of fuse gene in APL. The result shows the two types of isoforms of PML-RAR alpha have no significance in bcl-2 protein/mRNA expression, indexes of clinical prognosis. This shows that the classification of PML-RAR alpha isoform has no influence on clinical prognosis.

[Relationship between the expression of bcl-2 apoptosis and acute myelogenous leukemia prognosis].

In order to check the anti-apoptosis gene bcl-2 protein expression in 58 AML patients we used the immunohistochemical method, and analyse the relationship between bcl-2 protein and prognosis of AML. The result showed that the high expression of bcl-2 protein was related to poor clinical prognosis of AML. This shows that the expression of bcl-2 gene can serve as an index for the prognosis of AML.

Ion chromatographic determination of taurine in medicine, nutrient capsule and human urine with electrochemical detection.

A very simple and sensitive method for the determination of taurine by ion chromatography with electrochemical integrated pulsed amperometry is firstly described. Taurine was determined using 160 mmol/l NaOH as eluent and a Dionex CarboPac PA1 separation column (250x4 mm I.D.) without the interference with ten kinds of common amino acids. The peak area response of taurine was linear in the range 0.1-20 microg/ml, the detection limit was 0.034 microg/ml. The method has been applied successfully in the determination of taurine in medicinal granule, nutrient capsule and human urine. The content determined in medicinal granule is consistent with that marked by the manufacturer.