Cloning and expression of a novel partial cDNA related to hypertension.
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Animals↗
Biomedical subjects
G Dong
Publications and source records attributed to G Dong.
[Characters of semen of Curculigo plants under electron microscopic scanning].
The semens of five species of Curculigo were detected by electron microscopic scanning. The result shows that the surface characters of semens are useful in the identification of these species.
Magnoliopsida↗
[Stenosis of the common left iliac vein and acute deep vein thrombosis of the lower extremity].
OBJECTIVE: To study the relation ship between (DVT) acute deep vein thrombosis (DVT) of the lower extremity (DVT) and common left iliac vein compression by the common right iliac artery. METHOD: We reviewed 63 cases of DVT from September 1991 to March 1997. RESULTS: 43 cases operated on the left leg showed when the stenosis reached nearly or beyond half of the lumen of iliac vein, the number of DVT increased greatly. CONCLUSION: DVT will occur if the stricture of the left iliac vein reaches nearly or beyond half of the lumen. But there is no DVT because of collaterals. If the stricture of iliac vein exists, especially reaches nearly or beyond half of the iliac vein, preventive measures will be taken.
Adult↗
[The effects of fibronectin on the attachment and proliferation of periodontal ligament cells].
The purpose of this in vitro study was to observe and evaluate the effects of fibronectin(FN) on the growth of periodontal ligament cells(PDLC) on different root surfaces. Cell culture and scanning electronic microscopic techniques were used. The results showed that the growth pattern of PDLC on normal root surfaces were better than that of periodontitis. FN can significantly enhanced the attachment and proliferation of PDL cells on normal root surfaces. On the other hand, only the enhancement of attachment by FN was observed on the periodontitis root surfaces. It may be concluded that FN can promote the growth of PDLC and FN may play different effects on the attachment and proliferation of PDLC.
Cell Division↗
[A multiexponential model in cavity ring-down absorption spectroscopy].
A multiexponential model of cavity ring-down absorption spectroscopy is established for taking into account the laser linewidth effects. It is demonstrated that by fitting the logarithmic ring-down function to a truncated polynomial, absorption coefficent can be extracted from coefficients of the first as well as higher order terms of the fitted polynomial. The new fit model results in higher accuracy and broader dynamic range than the previous single exponential model.
English Abstract↗
Established immunity precludes adenovirus-mediated gene transfer in rat carotid arteries. Potential for immunosuppression and vector engineering to overcome barriers of immunity.
Preclinical arterial gene transfer studies with adenoviral vectors are typically performed in laboratory animals that lack immunity to adenovirus. However, human patients are likely to have prior exposures to adenovirus that might affect: (a) the success of arterial gene transfer; (b) the duration of recombinant gene expression; and (c) the likelihood of a destructive immune response to transduced cells. We confirmed a high prevalence (57%) in adult humans of neutralizing antibodies to adenovirus type 5. We then used a rat model to establish a central role for the immune system in determining the success as well as the duration of recombinant gene expression after adenovirus-mediated gene transfer into isolated arterial segments. Vector-mediated recombinant gene expression, which was successful in naive rats and prolonged by immunosuppression, was unsuccessful in the presence of established immunity to adenovirus. 4 d of immunosuppressive therapy permitted arterial gene transfer and expression in immune rats, but at decreased levels. Ultraviolet-irradiated adenoviral vectors, which mimic advanced-generation vectors (reduced viral gene expression and relatively preserved capsid function), were less immunogenic than were nonirradiated vectors. A primary exposure to ultraviolet-irradiated (but not nonirradiated) vectors permitted expression of a recombinant gene after redelivery of the same vector. In conclusion, arterial gene transfer with current type 5 adenoviral vectors is unlikely to result in significant levels of gene expression in the majority of humans. Both immunosuppression and further engineering of the vector genome to decrease expression of viral genes show promise in circumventing barriers to adenovirus-mediated arterial gene transfer.
Adenoviridae Infections↗
Purification and characterization of alkaline phosphatase from Thermotoga neapolitana.
A hyperthermophilic alkaline phosphatase was purified from Thermotoga neapolitana by heat treatment at 100 degrees C in the presence of Co2+ followed by ion-exchange and affinity chromatographies. The enzyme was purified 2,880-fold with 44% yield. The purified enzyme showed a single protein band of M(r) 45,000 on SDS-PAGE and an apparent molecular weight of 87,000 estimated by gel filtration chromatography. This suggested a homogenous dimer structure. The optimal pH and temperature for enzyme activity were 9.9 and 85 degrees C, respectively. Under optimal conditions, T. neapolitana alkaline phosphatase displayed 30% higher activity than calf intestine alkaline phosphatase did with p-nitrophenyl-phosphate as substrate. The hyperthermostable enzyme had a half-life of 238 min at 90 degrees C and K(m) and Vmax values of 183 microM and 1,352 U mg-1, respectively. Co2+ enhanced the enzyme activity, thermostability, and ligand affinity during column chromatography. The alkaline phosphatase was twice as active with Co2+ than with either Zn2+ or Mn2+ as the metal cofactor.
Alkaline Phosphatase↗
Cloning, sequencing, and expression of the gene encoding extracellular alpha-amylase from Pyrococcus furiosus and biochemical characterization of the recombinant enzyme.
The gene encoding the hyperthermophilic extracellular alpha-amylase from Pyrococcus furiosus was cloned by activity screening in Escherichia coli. The gene encoded a single 460-residue polypeptide chain. The polypeptide contained a 26-residue signal peptide, indicating that this Pyrococcus alpha-amylase was an extracellular enzyme. Unlike the P. furiosus intracellular alpha-amylase, this extracellular enzyme showed 45 to 56% similarity and 20 to 35% identity to other amylolytic enzymes of the alpha-amylase family and contained the four consensus regions characteristic of that enzyme family. The recombinant protein was a homodimer with a molecular weight of 100,000, as estimated by gel filtration. Both the dimer and monomer retained starch-degrading activity after extensive denaturation and migration on sodium dodecyl sulfate-polyacrylamide gels. The P. furiosus alpha-amylase was a liquefying enzyme with a specific activity of 3,900 U mg-1 at 98 degrees C. It was optimally active at 100 degrees C and pH 5.5 to 6.0 and did not require Ca2+ for activity or thermostability. With a half-life of 13 h at 98 degrees C, the P. furiosus enzyme was significantly more thermostable than the commercially available Bacillus licheniformis alpha-amylase (Taka-therm).
Amino Acid Sequence↗
Cloning, sequencing, and expression of the gene encoding amylopullulanase from Pyrococcus furiosus and biochemical characterization of the recombinant enzyme.
The gene encoding the Pyrococcus furiosus hyperthermophilic amylopullulanase (APU) was cloned, sequenced, and expressed in Escherichia coli. The gene encoded a single 827-residue polypeptide with a 26-residue signal peptide. The protein sequence had very low homology (17 to 21% identity) with other APUs and enzymes of the alpha-amylase family. In particular, none of the consensus regions present in the alpha-amylase family could be identified. P. furiosus APU showed similarity to three proteins, including the P. furiosus intracellular alpha-amylase and Dictyoglomus thermophilum alpha-amylase A. The mature protein had a molecular weight of 89,000. The recombinant P. furiosus APU remained folded after denaturation at temperatures of < or = 70 degrees C and showed an apparent molecular weight of 50,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Denaturating temperatures of above 100 degrees C were required for complete unfolding. The enzyme was extremely thermostable, with an optimal activity at 105 degrees C and pH 5.5. Ca2+ increased the enzyme activity, thermostability, and substrate affinity. The enzyme was highly resistant to chemical denaturing reagents, and its activity increased up to twofold in the presence of surfactants.
Amino Acid Sequence↗
Decreased expression of Wilms' tumor gene WT-1 and elevated expression of insulin growth factor-II (IGF-II) and type 1 IGF receptor genes in prostatic stromal cells from patients with benign prostatic hyperplasia.
Benign prostatic hyperplasia (BPH) is a common proliferative disorder of unknown etiology. We have previously documented that the insulin-like growth factor (IGF) axis is critical for prostate cell growth and is abnormal in BPH. The type 1 IGF receptor (IGF-1R) is constitutively expressed by most body tissues and plays a significant role in regulating cell proliferation, consistent with the role of its ligands (IGF-I and IGF-II) as important mitogenic factors. The Wilms' tumor gene product (WT-1) is a tumor suppressor that has been shown to be altered in rare kidney tumors and is known to regulate IGF-II and IGF-1R. We investigated the possibility that the expression of prostatic WT-1, IGF-1R, and IGF-II genes is altered in patients with BPH. We utilized primary cultures of prostatic stromal cells grown from normal (n = 9) and hyperplastic (n = 9) surgical specimens and analyzed WT-1, IGF-1R, and IGF-II messenger RNA levels. In all of the BPH cell strains, WT-1 expression (measured by RT-PCR and RNase protection assays) was strikingly lower than that found in normal strains (0-20% of normal, mean 14% of normal, P < 0.01). The expression of both the IGF-1R (300% of normal, P < 0.05) and IGF-II (1000% of normal, P < 0.01) messenger RNAs was higher in BPH strains as compared with normal strains. No changes were seen in stromal cell strains derived from prostatic adenocarcinoma. Thus, in cultured human prostatic stromal cell strains from patients with BPH, decreased WT-1 gene expression is associated with increases in the expression of the IGF-1R and IGF-II genes that are known transcriptional targets of WT-1. These findings indicate that reduced expression of the WT-1 tumor suppressor gene and elevated IGF-1R and IGF-II gene expression may be involved in the pathophysiology of prostatic hyperplasia, implying a new role for the Wilms' tumor gene in nonmalignant states.
Aged↗
Regional variability in the time course of TGF-beta 1 expression, cellular proliferation and extracellular matrix expansion following arterial injury.
Transforming growth factor-beta 1 (TGF-beta 1) has been variably associated with the regulation of cellular proliferation and extracellular matrix expansion after arterial injury. We tested these associations in vivo in the rat carotid injury model. At 0, 3, 7, 14 and 28 days following arterial balloon injury, regional expression of TGF-beta 1 mRNA was assessed using in situ hybridization and the results compared to measures of cellular proliferation and extracellular matrix expansion. Both the TGF-beta 1 concentration measured in culture media of explanted carotid arteries and the quantitative in situ hybridization signal for TGF-beta 1 arterial media and neointima were maximal at 14 days after balloon injury. However, medial cellular proliferation was maximal at 3 days whereas neointimal proliferation was maximal at 14 days and significantly greater than medial proliferation. Neointimal cell density declined significantly between 7 and 14 days, indicating the expansion of extracellular matrix; however, medial cell density was unchanged between 3 and 28 days after balloon injury. Thus, differences in the regional arterial wall relationships between the time course of cellular proliferation, extracellular matrix expansion and the level of TGF-beta 1 expression demonstrate in vivo variability in the response to TGF-beta 1.
Angioplasty, Balloon, Coronary↗
[The effects of transforming growth factor-beta and insulin on proliferation of human periodontal ligament cells].
AIM: To evaluate biological effects in vitro of transforming growth factor-beta (TGF-beta) and/or insulin on proliferation of the human periodontal ligament (PDL) cells. METHODS: Cell cultures and MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] colorimetric assay. RESULT: TGF-beta at concentrations of 0.1 to 10 micrograms/L and insulin at 10 to 1,000 U/L had significant proliferative effects on human PDL cells (P < 0.01). From the 3rd day, the significant proliferation of PDL cells by TGF-beta or insulin were observed (P < 0.01) and lasted to the 5th day. The effect of combination of TGF-beta with insulin was the most significant one (P < 0.01). CONCLUSION: TGF-beta and/or insulin may be valuable in promoting the regeneration of periodontal connective tissue.
Cell Division↗
[A study on location of poles of terminal leads of ventricular fractionated potentials].
To eliminate the electromyogram-noise-signal from recording ventricular fractionated potentials (VFP) of the terminal leads(TLs), the multi-locations of leads were used to record ECG and VFP in 36 cases of controls and 68 cases of coronary heart disease. It was shown that the electromyogram-noise-signal could be minimized when TLs located separately at the point of infraclavicular cross lines axillaries media and the point of upperinguen cross lines axillaries media, and the essential formation of the ECG and VFP signals could not be changed except the QRS potentials becoming higher.
Arm↗
Mutations identified in exon 7 of phenylalanine hydroxylase gene in Chinese.
Exon 7 of the phenylalanine hydroxylase (PAH) gene was analyzed in 45 children affected with classic phenylketonuria (PKU) from northern China by using PCR-single strand conformation polymorphism (PCR-SSCP) technique and DNA direct sequencing. Six missense mutations(i.e. R243Q, R241H, G247V, L249H, F2541 and G257V) and one silent mutation (V245v) were identified. The latter three missense mutations were demonstrated as novel mutations in comparison with the PAH mutation database. One missense mutation (R241H) was first documented in Chinese. Our results showed population and region differences in the PAH mutation distribution, and suggest that there is more than one founding population for PKU in China. The finding of novel mutations will enhence the molecular diagnosis of PKU.
Asian People↗
[Determination of glutamine in intestinal mucosa by pre-column derivatization/reversed-phase high performance liquid chromatography with fluorescence detection].
A high performance liquid chromatographic pre-column derivatization method with fluorescence detection for the analysis of glutamine (Gln) in rat intestinal mucosa is presented. Gln was derivatized with o-phthalaldehyde and 3-mercaptopropionic acid under an alkaline condition and separated by reversed-phase liquid chromatography on a Lichrosorb RP18 column (150 mm x 4.6 mm i.d., 5 microm). The mobile phase was consisted of 50 mmol/L phosphate buffer (pH 7.0)-acetonitrile (94:6, V/V) with a flow rate of 2.0 mL/min. The excited and emitted wavelength were selected at 230 nm and 389 nm respectively. The volume ratio of samples and derivatization reagent solution was 4:1 (V/V). The detection limit of Gln was 25 micromol/L (S/N= 3.5) and the regression equation was A = 16.9405C + 179.9339, r = 0.9996 at the linear range of 50-3200 micromol/L. The day-to-day deviation was 6.97% (n = 3) and the retention time of Gln was 3.158 min. This method is rapid, simple and highly sensitive, and has been applied to the determination of Gln in intestinal mucosa.
Animals↗
[Development of a SIRI-TOFMS and the application research progress].
The only one set of Sputter Initiated Resonance Ionization-Time of Flight Mass Spectrometer (SIRI-TOFMS) in China was designed and developed. A liquid gallium ion source with micron level diameter of beam, frame-style piezoeletric ceramic sub-micron sample manipulator, a secondary electron imaging system and a computer controlled precise targeting system as well as a real colour big screen image displaying system were added so as to improve its microanalysis capability. By means of this apparatus, trace analysis of Au in minerals and Cu in standard steel samples were carried out. The detection limit of them is 40ng/ and microg/g level, respectively. Aside from the above detections, other preliminary researches ere also performed.
English Abstract↗
[Novel mutations identified in exon 7 of phenylalanine hydroxylase gene in Chinese].
Exon 7 of the phenylalanine hydroxylase (PAH) gene was analyzed in 45 children affected with classic phenylketonuria (PKU) from northern China by using PCR-single strand conformation polymorphism (PCR-SSCP) technique and DNA direct sequencing. Six missense mutations (i.e. R243Q, R241H, G247V, L249H, P254I and G257V) and one silent mutation (V245V) were identified. The latter three missense mutations were demonstrated as novel mutations in comparison with the PAH mutation Database. One missense mutation (R241H) was first documented in Chinese. Our results showed population and regional differences in the PAH mutation distribution and suggest that there is more than one founding population for PKU in China. The finding of novel mutations will enhance our capability in molecular diagnosis of PKU.
Child↗
Genes differentially expressed with malignant transformation and metastatic tumor progression of murine squamous cell carcinoma.
Molecular changes occurring with tumor formation and metastasis need to be identified in order to define novel markers and targets for chemoprevention and therapy. Cell lines from a multistage model of murine squamous cell carcinoma were analyzed for differences in gene expression using mRNA differential display. mRNA was isolated from primary keratinocytes, an in vitro transformed keratinocyte line (Pam 212), and three metastatic cell lines derived from Pam 212 following tumor progression in vivo. cDNA was synthesized by reverse transcription and amplified by PCR using 72 primer combinations to screen and compare approximately 3,600 sequences. Five cDNAs with a differential expression pattern confirmed by Northern blot analysis were cloned and sequenced, revealing homology with known genes. The gene encoding tropomyosin alpha was preferentially expressed in primary keratinocytes; genes for tyrosine kinase Yes-associated protein (YAP65) and ribosomal protein L18a were preferentially expressed in transformed and metastatic tumor cell lines; and genes for the Gro-alpha family cytokine KC and antigen Sp17 exhibited increased expression in the three metastatic cell lines. The structure and function of the genes identified suggest that they may possibly be linked to cell shape and motility, signal transduction, protein synthesis, growth, granulocyte chemotaxis, and angiogenesis. This study demonstrates the ability of mRNA differential display to detect altered gene expression in this tumor progression model of murine squamous cell carcinoma, and the potential usefulness of this approach for identification of candidate genes as chemoprevention markers and targets.
Amino Acid Sequence↗