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Biomedical subjects

Q Fang

Publications and source records attributed to Q Fang.

At least 91 records · Page 5Linked to original sources
PubMed1994

V alpha gene usage in rheumatoid compared with osteoarthritic synovial tissue T cells.

While many investigators have examined V gene usage by the clonotypic T-cell receptor (TCR) in rheumatoid arthritis (RA) joints, few have reported on arthritic controls. We compared TCR alpha-chain V gene usage in knee synovial tissue specimens from 9 RA and 5 osteoarthritis (OA) patients. There was no significant difference in the number of V gene families used in RA compared with OA synovium. However, there was an increased prevalence of V alpha 28, V alpha 10, V alpha 17, and V alpha 18 and under representation of V alpha 15 in RA compared with OA synovium. Of these, V alpha 28 was also recently described by us as being present in RA synovial tissue early in the course of disease. V alpha 28 associated J region usage, and N-regional diversity was surveyed in T-cell receptors from additional rheumatoid synovial tissue T-cell populations and normal peripheral blood. Oligoclonality was observed in 6/10 rheumatoid specimens either by direct sequencing or where three or more molecular clones were sequenced, compared with 0/5 normal PBMCs. The oligoclonal populations included 2/3 cell lines stimulated with interleukin-2 (IL-2) alone. Several novel J regions were observed, with some recurrent residues observed at N-region positions. These data indicate an increased prevalence of certain TCR V region families in RA versus OA synovium, and suggest an antigen-driven expansion of V alpha 28-expressing T cells in RA synovium.

Amino Acid Sequence↗
PubMed1993

Human antibodies reactive with beta-amyloid protein in Alzheimer's disease.

Four human B cell lines established by Epstein-Barr viral transformation of B cells from a patient with a clinical diagnosis of Alzheimer's disease (AD) were found to secrete antibodies that react with plaques and cerebrovascular blood vessels in AD brain in a staining profile characteristic of beta-amyloid protein (beta-AP) in AD brain. Two of these antibodies were shown to be reactive with a rare plaque in a normal brain. In these studies, immunofluorescence and avidin-biotin complex immunoperoxidase methodology were used to determine antibody reaction, and thioflavine S was used to double label amyloid and neurofibrillary tangles. The four antibodies also reacted with neurons in normal and AD brain. Absorption studies, dot immunoblots, and enzyme-linked immunosorbent assays with beta-amyloid peptides 1-28 (beta-A1-28) and 1-40 (beta-A1-40) indicate the major determinant of the reactive epitope is located in the region of amino acids 1-28 of beta-AP. However, inhibition studies demonstrate a significant contribution to the antigenic determinant by the 29-40 region of the beta-A1-40. These antibodies represent the first human autoantibodies against beta-AP. The pathological significance of these autoantibodies is discussed.

Aged↗
PubMed1993

A phylogeny of New World Deltocephalus-like leafhopper genera based on mitochondrial 16S ribosomal DNA sequences.

The Deltocephalus-like leafhoppers are a group of grass-feeding insects that share common forms in the male genitalia, chiefly a fused aedeagal-connective structure. The group contains 27 New World genera. The phylogenetic relationships among these genera and other genera in the tribe are unclear because fusion of the aedeagus and connective, although clearly apomorphic, has occurred in both related and unrelated groups. An independent set of characters is required to test the hypothesis that the Deltocephalus-like genera are monophyletic and to derive relationships among these genera. We examined 562 sites in the 3'-end of the mitochondrial 16S ribosomal DNA from 21 species representing 19 New World genera. Eight species in closely related groups or tribes were also analyzed. The region was sequenced directly from PCR-amplified DNA of field-collected and museum-preserved dried specimens. The transversion rate was twice as high as the transition rate; however, 74% of these were adenine-thymine transversions. Analysis of secondary structure indicated that substitution rates were equal in stem and loop forming regions of the processed rRNA. Using either parsimony or distance analysis, derived phylogenies suggested that, with the exception of the genus Cabrulus, the Deltocephalus-like leafhoppers are genera within a monophyletic group.

Animals↗
PubMed1993

Conserved motifs in rheumatoid arthritis synovial tissue T-cell receptor beta chains.

Rheumatoid arthritis is genetically linked to major histocompatibility complex (MHC) molecules (HLA-DR4 and related molecules) and characterized pathologically by high levels of HLA-DR expression and infiltration of proliferative of synovial tissue with CD4+ T lymphocytes. T-lymphocyte activation is driven by specific signaling through polymorphic alpha/beta T-cell receptors (TCRs) that are reactive with antigen-MHC complexes present at the sites of inflammation. We are interested in characterizing rheumatoid TCRs molecularly to ascertain potential binding surfaces for antigen+MHC in synovial tissue. Accordingly, we have recently investigated the TCR alpha and beta chain heterogeneity in a series of 10 rheumatoid synovia obtained at the time of joint surgery. The most frequently detected V beta families were V beta 12, 14, and 17, each of which was found in 80% of specimens. We report here the molecular cloning and sequence analysis of 20 cloned V beta segments amplified with a V beta 14 family-specific TCR primer, and six cloned V beta segments amplified with a V beta 17 family-specific TCR primer from four rheumatoid synovia. Comparison with the data base revealed that these sequences belonged to the closely related V beta 3, V beta 14, and V beta 17 families. Dominant clones were apparent in two of the individuals by the presence of identical V-D-J regions, suggesting an antigen-driven process. Amino acid sequence analysis revealed a conserved motif in the putative fourth hypervariable region or CDR4. Molecular modeling of this epitope suggests that charged side chains are available for binding to ligand structures (e.g., antigen, MHC, or superantigen). We suggest this epitope may play a role in the molecular pathogenesis of rheumatoid arthritis.

Amino Acid Sequence↗
PubMed1993

[Hysterotomy and focal ectomy of malignant trophoblastic tumor].

Hysterotomy and focal ectomy were performed on 25 patients with malignant trophoblastic tumor during 1979-1991, owing to the patients and their husbands desiring strongly to have reproductive function preserved. Each of the foci of malignant trophoblastic tumor was found to be single nodule with 1-4 cm in diameter and located within the myometrium. The incision of uterus was sutured interuptly with "2-0" catgut. The pregnant rate was 80% among the patients who had received this surgical therapy. The results showed that hysterotomy and focal ectomy were an effective management for some patients without children.

Adult↗
PubMed1992

Specific DNA amplification utilizing the polymerase chain reaction and random oligonucleotide primers: application to the analysis of antigen receptor variable regions.

The polymerase chain reaction (PCR) allows rapid amplification of DNA of known sequence. In many situations, part of a genetic sequence is known, but adjacent sequences of interest are unknown. This is common in investigations of antigen receptor genes from B and T lymphocytes, which are composed of a constant region of known sequence and a variable region, for which the sequence may not be known. Herein is described a method to amplify DNA when sequence information is available for only one primer. This procedure utilizes a primer of known sequence in conjunction with a mixture of short random primers. Application of this method to the amplification of T-cell antigen receptor cDNA is described.

Arthritis, Rheumatoid↗
PubMed1992

Restricted heterogeneity of T cell receptor transcripts in rheumatoid synovium.

RA is characterized by massive proliferation of synovial tissue, accompanying infiltration of the tissue with CD4+ T lymphocytes, and a genetic linkage to the MHC antigen HLA-DR4. Since T cells are restricted by class II MHC molecules such as DR4, this suggests a direct role for these CD4+ cells in pathogenesis. To investigate T cell receptor (TCR) usage in RA, we used oligonucleotide primers specific for each of the major alpha and beta TCR subfamilies to amplify cDNA derived from whole synovium or synovial tissue T cell lines in a family-specific manner. Detection of amplified DNA was facilitated by utilizing oligonucleotide probes derived from the constant regions of the TCRs. The TCR repertoire present in the synovial T cell lines was quite heterogeneous, with an average of 15 alpha chains and 15.8 beta chains detected. When synovial tissue was analyzed, the predominant TCR subfamilies detected tended to be more restricted, with an average of 4.6 alpha chains and 8.6 beta chains detected. This compared with an average of six alpha chains and 12 beta chains in nonrheumatoid synovial samples. The average percentage of synovia positive per TCR beta family was significantly lower for RA versus non-RA specimens (46.1 vs 65.6%, P = 0.034). These findings indicate that while a polyclonal population of T cells is present in RA synovium, the predominant patterns of TCR transcript expression may be somewhat more restricted, suggesting that TCR-based therapy of RA is possible.

Arthritis, Rheumatoid↗
PubMed1992

Dystrophin cytochemistry in mdx mouse muscles injected with labeled normal myoblasts.

A new technique enables correlation of dystrophin expression with the location of donor versus host nuclei in the same sections of mdx mouse muscle injected with normal myoblasts. Myoblasts from C57BL/6J mice or from humans were labeled with 0.01% fluoro-gold (FG) in Dulbecco's Modified Eagles Medium (DMEM) for 16 h at 37 degrees C before myoblast transfer. About 3 x 10(4) myoblasts were injected into the quadriceps muscles of mdx mice immunosuppressed with cyclosporine A (CsA). At 11, 21, or 25 days after myoblast transfer, injected muscles were dissected out and sectioned. These mouse sections were processed for dystrophin and then labeled with a fluorescent nucleus counterstain, 5 micrograms% Hoechst 33342 in phosphate-buffered saline (PBS), for 10 min at room temperature. Fluoro-gold labeling corresponding with Hoechst 33342 staining indicated survival of normal nuclei in dystrophic muscle. Dystrophin was found in the sarcolemma of myofibers containing FG-labeled nuclei but not of myofibers containing only Hoechst 33342-labeled nuclei. Control muscle samples showed neither FG labeling nor dystrophin. This study demonstrates that the donor human and mouse myoblasts survived and developed in host mouse muscles for at least 25 days after myoblast transfer, and that the localization of their normal nuclei correlates with dystrophin expression in muscle fibers of immunosuppressed mdx host mice.

Animals↗
PubMed1992

Feasibility, safety, and efficacy of myoblast transfer therapy on Duchenne muscular dystrophy boys.

Five billion normal myoblasts were injected into each of 21 Duchenne muscular dystrophy (DMD) boys aged 6-14 yr to assess the feasibility, safety, and efficacy of the Phase II myoblast transfer therapy (MTT). The Phase II study was designed to strengthen muscles of both lower limbs. Forty-eight intramuscular injections transferred the myoblasts into 22 major muscles at 55.6 x 10(6)/mL in 10 min under general anesthesia. Eleven boys had received 8 million myoblasts each 1 yr ago in the Phase I MTT. In the Phase II study, eight of them had their myoblasts subcultured from reserves frozen 1 yr ago. The donor myoblasts for each of the remaining boys were cultured from satellite cells derived from a 1-g muscle biopsy of a normal male who might or might not be histocompatible with the recipient. The immunosuppressant cyclosporine (Cy) is being administered to recipients for 6 mo after MTT to facilitate donor cell survival. There was no evidence of an adverse reaction to MTT or Cy as determined by serial laboratory evaluations including electrolytes, creatinine, and urea. Early objective functional tests using the KinCom Robotic Dynamometer were conducted on 13 subjects aged 6 to 13 before MTT and at 3 mo after MTT. Of the 69 muscle groups (knee extensors, knee flexors, plantar flexors) tested for isometric force generation in these subjects, 43% showed mean increase of 41.3% +/- 5.9 SEM, 38% showed no change, and 19% showed continuous force reduction of 23.4% +/- 3.1 SEM. The remaining subjects await the 3-mo post-MTT evaluation. The results indicate that 1) MTT is safe; 2) MTT increases muscle strength in DMD: 81% of the muscles tested showed either increase in strength or did not show continuous loss of strength; 3) more than 5 billion myoblasts can be cultured from 1 g normal muscle biopsy, providing unprecedented numbers of cells for MTT; 4) myoblasts, frozen over 1 yr, retain the ability to proliferate from 10 million to 5 billion, and to form normal myofibers; 5) injections of 5 billion myoblasts have not provoked any immunological rejection symptoms in the Phase II subjects, 11 of whom received 8 million myoblasts in the Phase I MTT a year ago; 6) it is safe to perform multiple injections of myoblasts into lower limb muscles without formation of emboli; and 7) donor cell rejection by the recipient can be prevented with Cy when properly managed.

Adolescent↗
PubMed1992

Construction of an Rb gene expression plasmid.

We obtained an 844 bp Bg1II fragment from an Rb cDNA clone and inserted it into the expression vector pWR-13 to construct an Rb gene expression plasmid. When the Rb Bg1II fragment was fused in-frame into pWR-13, it was operated by a Lac Z promoter and produced a fusion protein which consisted of expressed Rb protein and a small peptide from Lac Z. The recombinants were transformed into E. coli with the CaCl2 method, screened by in situ hybridization, and restriction mapped. Total cellular protein of transformed clones was analyzed by SDS-PAGE and Commassie blue staining. The sense clones showed a unique band at 28,000. On Western blot, this band specifically reacted with 125I-labelled antibody against synthetic Rb peptide. This protein comprised more than 5% of total bacterial protein.

Amino Acid Sequence↗
PubMed1992

Clinical manifestations and prevalence of different types of supraventricular tachycardia among Chinese.

Sixty-nine cases of paroxysmal supraventricular tachycardia (PSVT) were submitted to electrophysiological studies. The prevalence of different types of PSVT is as follows: 49% of the cases had atrioventricular reentry tachycardia (AV-RT), 14.5% atrioventricular nodal reentry tachycardia (AVN-RT), 28% AV + AVN - RT, and 8.5% other types. Altogether 77% in this group had atrioventricular accessory pathway, of which half are concealed pathway. Electrocardiographic characteristics during tachycardia give hints to diagnosis: (1) AV - RT has the fastest heart rate, with 62% 200 BPM or more, and 82% more than 188 BPM; (2) retrograde P waves can be detected in 64% of cases with AV bypass; (3) 50% of AV - RT shows QRS of bundle branch block pattern. Since accessory pathways are present in the majority of PSVT patients in China, the importance of treatment is stressed.

China↗