Biomedical subjects

G Feng

Publications and source records attributed to G Feng.

At least 109 records · Page 6Linked to original sources

PubMed1998

[The establishment, freezing-state preservation and resuscitation of retinoblastoma SO-Rb70 cell line].

PURPOSE: To establish a new cell line of human retinoblastoma (Rb), frozen and resuscitated to provide a Rb cell reservoir for clinical and experimental research. METHODS: The screening cells were washed and blown directly without centrifugation and then inoculated. Based on the cell-frozen principle, the Rb cell line was collected and established, then frozen in liquid nitrogen and resuscitated. RESULTS: SO-Rb70 cell line was established successfully. 8 groups of cells were frozen. The resuscitation experiment was carried out for 6 times, 4 times being successful. CONCLUSION: The preserved human Rb cells in liquid nitrogen grew normally after resuscitation with the maintenance of previous bioactivities and characteristics. This research helped to provide abundant material for further study of Rb.

Cell Line↗

PubMed1997

Development of a new kind of dual modulated QCM biosensor.

To distinguish the mass loading effect from the total frequency change is a problem in the application of a quartz crystal microbalance (QCM) biosensor in the liquid phase. Based on the characteristic damping theory, this paper proposes a new method of dual modulation to solve this problem. Using polyethyleneimine to immobilize anti-SE (staphylococcin enterotoxin) antibody (C2 type), a dual modulated QCM SE biosensor was developed and the experiment proved that it has little cross-reaction with B-type SE. The measuring curve of the sensor was also determined through experiment.

Biosensing Techniques↗

PubMed1997

epsilon-Sarcoglycan, a broadly expressed homologue of the gene mutated in limb-girdle muscular dystrophy 2D.

The sarcoglycans are transmembrane components of the dystrophin-glycoprotein complex, which links the cytoskeleton to the extracellular matrix in adult muscle fibers. Mutations in all four known sarcoglycan genes (alpha, beta, gamma, and delta) have been found in humans with limb-girdle muscular dystrophy. We have identified a novel protein, epsilon-sarcoglycan, that shares 44% amino acid identity with alpha-sarcoglycan (adhalin). We show that epsilon-sarcoglycan is a membrane-associated glycoprotein and document its expression by Northern blotting, immunoblotting, and immunofluorescence. In contrast to alpha-delta sarcoglycans, which are expressed predominantly or exclusively in striated muscle, epsilon-sarcoglycan is broadly distributed in muscle and nonmuscle cells of both embryos and adults. These results raise the possibility that sarcoglycan-containing complexes mediate membrane-matrix interactions in many cell types.

Amino Acid Sequence↗

PubMed1997

Mismatch repair protein MutL becomes limiting during stationary-phase mutation.

Postsynthesis mismatch repair is an important contributor to mutation avoidance and genomic stability in bacteria, yeast, and humans. Regulation of its activity would allow organisms to regulate their ability to evolve. That mismatch repair might be down-regulated in stationary-phase Escherichia coli was suggested by the sequence spectrum of some stationary-phase ("adaptive") mutations and by the observations that MutS and MutH levels decline during stationary phase. We report that overproduction of MutL inhibits mutation in stationary phase but not during growth. MutS overproduction has no such effect, and MutL overproduction does not prevent stationary-phase decline of either MutS or MutH. These results imply that MutS and MutH decline to levels appropriate for the decreased DNA synthesis in stationary phase, whereas functional MutL is limiting for mismatch repair specifically during stationary phase. Modulation of mutation rate and genetic stability in response to environmental or developmental cues, such as stationary phase and stress, could be important in evolution, development, microbial pathogenicity, and the origins of cancer.

Adenosine Triphosphatases↗

PubMed1997

Negative regulation of mutS and mutH repair gene expression by the Hfq and RpoS global regulators of Escherichia coli K-12.

The MutS, MutL, and MutH proteins play major roles in several DNA repair pathways. We previously reported that the cellular amounts of MutS and MutH decreased by as much as 10-fold in stationary-phase cultures. Consequently, we tested whether the amounts of MutS, MutL, and MutH were regulated by two global regulators, RpoS (sigma38) and Hfq (HF-I [putative RNA chaperone]), which are involved in stationary-phase transition. We report here that mutations in hfq and rpoS reversed the stationary-phase down-regulation of the amounts of MutS and MutH. hfq regulation of the amount of MutS in stationary-phase cultures was mediated by RpoS-dependent and -independent mechanisms, whereas hfq regulation of the amount of MutH was mediated only through RpoS. Consistent with this interpretation, the amount of MutS but not MutH was regulated by Hfq, but not RpoS, in exponentially growing cells. The amount of MutL remained unchanged in rpoS, hfq-1, and rpoS+, hfq+ strains in exponentially growing and stationary-phase cultures and served as a control. The beta-galactosidase activities of single-copy mutS-lacZ operon and gene fusions suggested that hfq regulates mutS posttranscriptionally in exponentially growing cultures. RNase T2 protection assays revealed increased amounts of mutS transcript that are attributed to increased mutS transcript stability in hfq-1 mutants. Lack of Hfq also increased the amounts and stabilities of transcripts initiated from P(miaA) and P1hfqHS, two of the promoters for hfq, suggesting autoregulation, but did not change the half-life of bulk mRNA. These results suggest that the amounts of MutS and MutH may be adjusted in cells subjected to different stress conditions by an RpoS-dependent mechanism. In addition, Hfq directly or indirectly regulates several genes, including mutS, hfq, and miaA, by an RpoS-independent mechanism that destabilizes transcripts.

Adenosine Triphosphatases↗

PubMed1997

[Serial modifications of bacon's pull through resection for low rectal cancer].

Four modifications of Bacon's pull-thru resection were undertaken to improve postoperative defecatory control and to avoid the 2nd stage resection of the pull-thru colon stump. 54 cases of modified Bacon's rectal resection (9.64% of sphincter saving resections) were performed between 1954-1989. The 1st modification (1954) preserves the levator ani and the intact anorectal ring. The 2nd modification (1964) preserves the dentate margin and anal transitional zone (acute anal sensation), thus greatly improving defecatory control. The 3rd modification (1980) simplified intraanal resective procedure. The 4th modification (1991) was to encircle and ligate the distal end of colon over a sterilized corrugated intraluminal splinting tube (The other end of which is already connected to a long latex tube prior to operation). The latex tube was pulled out thru anus, until the colonic ligature reaches the Dentate margin, to substitute for colonic pull-thru and to divert feces during & after operation, 4-8 fine stitches approximate the colon wall 1 cm proximal to the ligature, to the cut edge of anal mucosa. The colon wall distal to the ligature sloughed in 7-10 days, the proximal colon has partially healed to the raw areas of anal canal without infection. The 2nd stage colon stump resection is thus obviated and hospitalization shortened. The postoperative anal function was good in 86.66%. Modified Bacon's operation is indicated for very low rectal cancer when the rectal remnant above levator ani after adequate resection is less than 1 cm which is difficult for intraabdominal anastomosis. It extends the scope of sphincter-saving operation. It is a good substitute for Park's coloanal anastomosis.

Defecation↗

PubMed1997

[Clinical and histological findings of retrocorneal membrane after keratoplasty].

PURPOSE: To observe the clinicopathologic changes of retrocorneal membrane after keratoplasty. METHODS: Light microscope and histochemical technique were used. RESULTS: The clinicopathologic review of 108 unsuccessful keratoplasties revealed the presence of 45 retrocorneal membrane, which seems to depend on the wound abnormality, rejection of corneal transplantion, epithelium in growth, hyphema, and iris synechias. CONCLUSION: The retrocorneal membrane derives from the host corneal stromal keratocytes and the metaplasia of corneal endothelial cells and monocytes.

Corneal Diseases↗

PubMed1996

Cloning and functional characterization of a novel dopamine receptor from Drosophila melanogaster.

A cDNA clone is described that encodes a novel G-protein-coupled dopamine receptor (DopR99B) expressed in Drosophila heads. The DopR99B receptor maps to 99B3-5, close to the position of the octopamine/tyramine receptor gene at 99A10-B1, suggesting that the two may be related through a gene duplication. Agonist stimulation of DopR99B receptors expressed in Xenopus oocytes increased intracellular Ca2+ levels monitored as changes in an endogenous inward Ca2+-dependent chloride current. In addition to initiating this intracellular Ca2+ signal, stimulation of DopR99B increased cAMP levels. The rank order of potency of agonists in stimulating the chloride current is: dopamine > norepinephrine > epinephrine > tyramine. Octopamine and 5-hydroxytryptamine are not active (< 100 microM). This pharmacological profile plus the second-messenger coupling pattern suggest that the DopR99B receptor is a D1-like dopamine receptor. However, the hydrophobic core region of the DopR99B receptor shows almost equal amino acid sequence identity (40-48%) with vertebrate serotonergic, alpha 1- and beta-adrenergic, and D1-like and D2-like dopaminergic receptors. Thus, this Drosophila receptor defines a novel structural class of dopamine receptors. Because DopR99B is the second dopamine receptor cloned from Drosophila, this work establishes dopamine receptor diversity in a system amenable to genetic dissection.

Amino Acid Sequence↗

PubMed1996

Amino acid substitutions in the two largest subunits of Escherichia coli RNA polymerase that suppress a defective Rho termination factor affect different parts of the transcription complex.

Among the earliest rpoBC mutations identified are three suppressors of the conditional lethal rho allele, rho201. These three mutations are of particular interest because, unlike rpoB8, they do not increase termination at all rho-dependent and rho-independent terminators. rpoB211 and rpoB212 both change Asn-1072 to His in conserved region H of rpoB (betaN1072H), whereas rpoC214 changes Arg-352 to Cys in conserved region C of rpoC (beta'R352C). Both substitutions significantly reduce the overall rate of transcript elongation in vitro relative to wild-type RNA polymerase; however, they probably slow elongation for different reasons. The nucleotide triphosphate concentrations required at the T7 A1 promoter for both abortive trinucleotide synthesis and for promoter escape are much greater for betaN1072H. In contrast, beta'R352C and two adjacent substitutions (beta'G351S and beta'S350F), but not betaN1072H, formed open complexes of greatly reduced stability. The sequence in this region of beta' modestly resembles a region of Escherichia coli DNA polymerase I that contacts the phosphate backbone of DNA in co-crystals. Core determinants affecting open complex formation do not reside exclusively in beta', however, since the Rifr mutation rpoB2 in beta also dramatically destabilized open complexes. We suggest that the principal defects of the two Rho-suppressing substitutions may differ, perhaps reflecting a greater role of beta region H in nucleoside triphosphate-binding and nucleotide addition and of beta' region C in contacts to the DNA strands that could be important for translocation. Although both probably suppress rho201 by slowing RNA chain elongation, these differences may lead to terminator specificity that depends on the rate-limiting step at different sites.

Amino Acid Sequence↗

PubMed1996

[Comparative study on the long-term effect of permanent embolization of hepatic artery with Bletilla striata in patients with primary liver cancer].

From May, 1990 to Sept., 1993 106 cases of primary liver cancer were treated by temporary or permanent hepatic arterial embolization with Bletilla striata powders (permanent, 56 cases) or gelfoam powders (temporary, 50 cases) under controlled technical conditions, in term of degree of tumor necrosis, revascularization of tumor, and long-term effect, the results of Bletilla striata group were better as compared with those of gelfoam group. The 1, 2 and 3 year survival rates were 44.9%, 33.6% and 33.6% in Beltilla striata group while the rates were 48.9%, 31.1% and 16.0% in gelfoam group, suggesting that Bletilla striata is superior to gelfoam as an embolizing agent for hepatic carcinoma.

Antineoplastic Agents, Phytogenic↗

PubMed1996

Renal angiomyolipoma: diagnosis with B-ultrasonography, CT scanning, DSA and its interventional treatment.

From 1989, 15 cases of renal angiomyolipoma (AML) have been diagnosed by ultrasonography. CT scanning and digital subtraction angiography (DSA) at our hospital. In 8 patients with uneven hyperechoes on B-mode ultrasonography (B-US) (8/15) and 7 with low density of fat on CT scanning (7/12) accurate diagnosis was established preoperatively. DSA revealed the "berry-like" pseudoaneurysms in the arterial phase (14 cases), the defined lucent area in the nephrogram phase (10 cases) and the "onion-peel appearances" during venous phases (8 cases), correct diagnosis was achieved in all patients. 8 cases were surgically treated and 7 treated by subselective embolization of renal artery. Effects in all cases were good. The diagnostic value of B-US, CT scanning, DSA and interventional treatment of AML was discussed. It was believed that the diagnosis with DSA was a technique with high specificity, and embolization therapy was simple and effective for AML.

Adult↗

PubMed1996

Some acoustic features of nasal and nasalized vowels: a target for vowel nasalization.

In order to characterize acoustic properties of nasal and nasalized vowels, these sounds will be considered as a dynamic trend from an oral configuration toward an [n]-like configuration. The latter can be viewed as a target for vowel nasalization. This target corresponds to the pharyngonasal tract and it can be modeled, with some simplifications, by a single tract without any parallel paths. Thus the first two resonance frequencies (at about 300 and 1000 Hz) characterize this target well. A series of measurements has been carried out in order to describe the acoustic characteristics of the target. Measured transfer functions confirm the resonator nature of the low-frequency peak. The introduction of such a target allows the conception of the nasal vowels as a trend beginning with a simple configuration, which is terminated in the same manner, so allowing the complex nasal phenomena to be bounded. A complete study of pole-zero evolutions for the nasalization of the 11 French vowels is presented. It allows the proposition of a common strategy for the nasalization of all vowels, so a true nasal vowel can be placed in this nasalization frame. The measured transfer functions for several French nasal vowels are also given.

France↗

PubMed1996

Transcription of the mutL repair, miaA tRNA modification, hfq pleiotropic regulator, and hflA region protease genes of Escherichia coli K-12 from clustered Esigma32-specific promoters during heat shock.

The amiB-mutL-miaA-hfq-hflX-hflK-hflC superoperon of Escherichia coli contains genes that are important for diverse cellular functions, including DNA mismatch repair (mutL), tRNA modification (miaA), pleiotropic regulation (hfq), and proteolysis (hflX-hflK-hflC). We show that this superoperon contains three E simga(32)-dependent heat shock promoters, P(mutL)HS,P(miaA)HS, and P1(hfq)HS, in addition to four E sigma(70)-dependent promoters, P(mutL), P(miaA), P2(hfq), and P3(hfq). Transcripts from P(mutL)HS and P(miaA)HS were most prominent in vivo during extreme heat shock (50 degrees C), whereas P1(hfq)HS transcripts were detectable under nonshock conditions and increased significantly after heat shock at 50 degrees C. The P(mutL)HS, P(miaA)HS, and P1(hfq)HS transcripts were not detected in an rpoH null mutant. All three promoters were transcribed by E sigma (32) in vitro at 37 degrees C and contain -35 and -10 regions that resemble the E sigma(32) consensus. In experiments to assess the possible physiological relevance of the P(mutL)HS and P(miaA)HS promoters, we found that E. coli prototrophic strain MG 1655 increased in cell mass and remained nearly 100% viable for several hours at 50 degrees C in enriched media. In these cells, a significant fraction of mutL and hfq-hflA region transcripts were from P(mutL)HS and P1(hfq)HS, respectively, and the amounts of the miaA, hfq, hflX, hflK, and hflC transcripts increased in comparison with those in nonstressed cells. The cellular amounts of MutL and the hfq gene product (HF-I protein) were maintained during heat shock at 44 or 50 degrees C. Consistent with their expression patterns, miaA and hfq were essential for growth and viability, respectively, at temperatures of 45 degrees C and above. Together, these results suggest that there is a class of E sigma(32) promoters that functions mainly at high temperatures to ensure E. coli function and survival.

Adenosine Triphosphatases↗

PubMed1996

Depletion of the cellular amounts of the MutS and MutH methyl-directed mismatch repair proteins in stationary-phase Escherichia coli K-12 cells.

The MutL, MutS, and MutH proteins mediate methyl-directed mismatch (MDM) repair and help to maintain chromosome stability in Escherichia coli. We determined the amounts of the MDM repair proteins in exponentially growing, stationary-phase, and nutrient-starved bacteria by quantitative Western immunoblotting. Extracts of null mutants containing various amounts of purified MDM repair proteins were used as quantitation standards. In bacteria growing exponentially in enriched minimal salts-glucose medium, about 113 MutL dimers, 186 MutS dimers, and 135 MutH monomers were present per cell. Calculations with the in vitro dissociation constants of MutS binding to different mismatches suggested that MutS is not present in excess, and may be nearly limiting in some cases, for MDM repair in exponentially growing cells. Remarkably, when bacteria entered late stationary phase or were deprived of a utilizable carbon source for several days, the cellular amount of MutS dropped at least 10-fold and became barely detectable by the methods used. In contrast, the amount of MutH dropped only about threefold and the amount of MutL remained essentially constant in late-stationary-phase and carbon-starved cells compared with those in exponentially growing bacteria. RNase T2 protection assays showed that the amounts of mutS, mutH, and mutL, but not miaA, transcripts decreased to undetectable levels in late-stationary-phase cells. These results suggested that depletion of MutS in nutritionally stressed cells was possibly caused by the relative instability of MutS compared with MutL and MutH. Our findings suggest that the MDM repair capacity is repressed in nutritionally stressed bacteria and correlate with conclusions from recent studies of adaptive mutagenesis. On the other hand, we did not detect induction of MutS or MutL in cells containing stable mismatches in multicopy single-stranded DNA encoded by bacterial retrons.

Adenosine Triphosphatases↗

PubMed1996

A 60-kilodalton protein in rat hepatoma cells overexpressing insulin receptor was tyrosine phosphorylated and associated with Syp, phophatidylinositol 3-kinase, and Grb2 in an insulin-dependent manner.

Tyrosine phosphorylation of cellular proteins is an early and key step after activation of the insulin receptor kinase (IRK). The study of the properties of these proteins should contribute to our understanding of insulin action. In rat hepatoma cells overexpressing human insulin receptors (HTC-IR), insulin treatment resulted in rapid tyrosine phosphorylation of proteins of 180, 94, 68, and 60 kDa. When lysates from insulin-treated cells were immunoprecipitated with anti-Syp antibody, subsequent immunoblotting identified p65 and p68, which reacted with anti-Syp, and p6O and p68, which reacted with antiphosphotyrosine antibody. Thus, insulin treatment yielded tyrosine phosphorylation of both Syp and a Syp-associated p6O molecule. When lysates from insulin-treated cells were adsorbed with a glutathione S-transferase (GST)-Syp-Src homology-2 (SH2) fusion protein, tyrosine- phosphorylated p6O was sequestered. After subjecting lysates to SDS-PAGE, the GST-SypSH2 fusion protein was found to bind to p18O, p94, and p6O. Thus, Syp associates directly with a 60-kDa IRK substrate via its SH2 domains. Syp-associated p6O differed from the 60- to 62-kDa proteins, associating with ras guanosine triphosphatase-activating protein, which also underwent modest tyrosine phosphorylation in response to insulin. Preadsorption of cell lystates with antibody against the 85-kDa subunit (p85) of phosphatidylinositol 3-kinase substantially reduced the amount of p60 subsequently immunoprecipitated by anti-Syp. Thus, p60 associates with both Syp and p85. The amount of tyrosine-phosphorylated p60 exceeded that of p180 in anti-Syp immunoprecipitates, whereas their proportion was comparable in anti-p85 immunoprecipitates. Grb2 was also observed in the anti-Syp immunoprecipitates. When lysates from insulin-treated cells were adsorbed with GST-p85SH2 domains or GST-Grb2, the subsequent eluates contained tyrosine-phosphorylated p60, as determined by immunoblotting with antiphosphotyrosine. Membrane binding assays using GST fusion proteins showed that these associations were direct. Studies in rat liver, muscle, and adipose tissue identified insulin-dependent association of Syp, Grb2, and p85 with tyrosine-phosphorylated p60 in adipose tissue only. We conclude that insulin treatment of HTC-IR cells and rat adipose tissue results in the tyrosine phosphorylation of p60, which might participate in the recruitment of downstream effectors involved in insulin signal transduction.

Adaptor Proteins, Signal Transducing↗

PubMed1996

[Observation of the invasion of retinoblastoma cells to ocular tissues in vitro].

PURPOSE: To study the invasion of retinoblastoma cells to ocular tissues. METHODS: The SO-Rb50 cells of retinoblastoma and various ocular tissue cells were co-cultured. The morphological change of the SO-Rb50 cells adherent to ocular tissue cells was observed. RESULTS: SO-Rb50 cells can adhere to the various ocular tissues. The adherent ability of SO-Rb50 cell to different ocular tissue cells was different. Tumor cells could be adhesive to keratocyte, scleral fibrocyte, the fibrocyte and the melanocyte of iris and choroid, the epithelial cells of lens, and the astrocyte of optic nerve and grow, but can not be adhesive to the corneal epithelial cell and retinal pigment epithelial cell. CONCLUSIONS: We consider that the interactions of tumor cells with host cells, as with extracellular matrix, also play an important role in the selective growth of organ and tissue of tumor cell metastasis. Retinoblastoma cells can not adhere to retinal pigment epithelial cell, which may serve as a barrier to obstruct the invasion of tumor cell to the choroid. Being adherent to the glial cells of optic nerve, tumor cells grow in flat and the shape of tumor cell changes, which may be the cause of tumor cell migrate rapidly and grow in the brain while the optic nerve was invaded by tumor cells.

Adult↗

PubMed1996

[New use of Bletilla striata as embolizing agent in the intervention treatment of hepatic carcinoma].

A Chinese herb, Bletilla striata, was used as embolizing agent in order to improve the therapeutic results of intervention treatment of liver cancer. From October 1991 to January 1995, 56 cases of hepatic carcinoma were treated with Bletilla striata by hepatic artery embolization, with conventional gelform embolization in 50 cases as control. Patients were followed-up for 10-48 months. Embolization with Bletilla striata led to extensive and permanent vascular obstruction, accompanied with marked shrinkage of tumor size and significant decrease in serum AFP levels. Collaterals were few in number and collateral circulation was established late so that the treatment intervals could be prolonged, with an average of 7 months. The 1-, 2-, and 3-year survival rate was 81.9%, 44.9% and 33.6%, respectively, with a median survival time of 19.8 months. All the clinical parameters were better than those treated with conventional gelform embolization. The results indicate that Bletilla striata is an ideal vascular embolizing agent.

Adult↗

PubMed1996

[Observation of the invasion of retinoblastoma cells to the corneal epithelial cells and keratocytes in culture].

OBJECTIVE: To study the invasive ability of retinoblastoma cells into the corneal epithelial cells and keratocytes in culture. METHODS: SO-Rb50 cells of retinoblastoma, corneal epithelial cells and keratocytes were co-cultured. The morphological changes of SO-Rb50 cells adherent to the corneal epithelial cells and keratocytes were observed. The distribution of proteoglycans in the SO-Rb50 cells, corneal cells and keratocytes was analyzed by histochemical method and electron microscopy. RESULTS: SO-Rb50 cells could adhere to the keratocytes, but did not adhere to the corneal epithelial cells. There are common hyaluronidase-resistant and chondroitinase-resistant proteoglycans on the surface of SO-Rb50 cells and corneal epithelial cells. Chondroitin is present on the surface of keratocytes. CONCLUSION: It is considered that the cell surface proteoglycans may play an important role in mediating SO-Rb50 tumor cell adhesion and non-adhesion to normal cells.

Adult↗