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Biomedical subjects

Q Sun

Publications and source records attributed to Q Sun.

At least 181 records · Page 10Linked to original sources

99Tcm-MIBI mammoscintigraphy of breast masses: early and delayed imaging.

Forty patients with breast mass and 10 female controls underwent double-phase (15 min and 3 h) 99Tcm-sestamibi (MIBI) imaging. Another four post-mastectomy patients with suspected metastases underwent total body imaging at 15 min. The 99Tcm-MIBI images of all 10 female controls showed symmetrical homogeneous radioactivity in both breasts. A focal area of increased 99Tcm-MIBI uptake in the breast tumour is considered as positive. Eleven breasts of 10 patients underwent total mastectomies with lymph node dissection, with one patient having bilateral mastectomy. These 11 breast tumours were confirmed to be infiltrating ductal carcinoma. All 11 breast carcinomas had positive early and delayed phase results with the exception of one patient who had only a focal increased uptake in early phase which happened to be the smallest size of carcinoma. Focal areas of positive MIBI uptake in the axillae in 4 of 10 patients were confirmed to be metastatic carcinoma of the breast. One patient with breast carcinoma underwent mastectomy whose 99Tcm-MIBI imaging was negative. Twelve patients undergoing excisional biopsy or lumpectomy of breast resulted in 7 fibroadenomas, 2 papillomas, 1 abscess, 1 granuloma and 1 chronic inflammation. Four of 7 fibroadenomas having hypercellularity histopathologically exhibited a focal MIBI uptake on double-phase imaging. One granulomatous disease had positive early phase but faded on the delayed image. The patients with absence of MIBI uptake included 3 fibroadenomas, 2 papillomas, 1 abscess, 1 chronic inflammation and 17 fibrous hyperplasia.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

In vivo association of Grb2 with pp116, a substrate of the T cell antigen receptor-activated protein tyrosine kinase.

Numerous recent studies have implicated the src homology 2 and 3 domain-containing protein, Grb2, in coupling protein tyrosine kinase signaling pathways with the Ras signaling pathway. Ligation of the T cell antigen receptor results in the activation of both a PTK, and Ras; therefore, we investigated whether Grb2 may serve a similar function in T cells. Here we report that a GST/Grb2 fusion protein associates with several tyrosine phosphoproteins from lysates of T cell antigen receptor-stimulated Jurkat T cells. Two of these proteins, pp36 and pp116, bind to the Grb2 fusion protein with high affinity. Through the use of mutated Grb2 fusion proteins, we demonstrate that pp116 binds the amino-terminal src homology 3 domain of Grb2, the same domain of Grb2 thought to be primarily responsible for its interaction with SOS. We demonstrate further that pp116 associates with Grb2 in vivo, and we provide evidence that in the Jurkat T cell line Grb2 may exist complexed with either pp116 or with SOS.

Adaptor Proteins, Signal Transducing↗

Characterization of the release of cholecystokinin from a murine neuroendocrine tumor cell line, STC-1.

The murine neuroendocrine cell line, STC-1, was found to contain 296.8 +/- 1.8 fmol of cholecystokinin-like immunoreactivity (CCK-LI) per mg cell protein. Immunocytochemical stain of STC-1 cells maintained in monolayer culture indicated that CCK-LI activity was present in 93% of the cells. Analysis by reverse-phase high-performance liquid chromatography indicated that STC-1 cells contained CCK-8 and an unidentified form as the predominant storage form. form. However, only CCK-8 was released into the culture medium upon stimulation by various secretagogues. The release of CCK-LI from STC-1 cells was stimulated by dibutyryl cAMP, forskolin, KCl, A23187, 4 beta-phorbol 12-myristate 13-acetate and luminal stimulants, e.g., sodium oleate, L-tryptophan, camostat and plaunotol. The release of CCK-LI from STC-1 cells was also stimulated by a neuropeptide, bombesin. The stimulatory effects of most of these agents were dose dependent. The stimulatory effects of dibutyryl cAMP, forskolin, and plaunotol were potentiated by 3-isobutyl-1-methyl xanthine, while that of camostat was not. The results obtained in this study indicate that the release of CCK from STC-1 cells shares the same characteristics of CCK release as from the CCK-secreting cells of the intestinal mucosa observed both in the dog and the rat in vitro and in vivo. Thus, the cellular mechanism of CCK release which appears to be cAMP- and Ca(2+)-dependent may be modulated by cellular protein kinase C activity. The STC-1 cell appears to be a suitable model for studying the mechanism of CCK release.

Animals↗

A purine-rich exon sequence enhances alternative splicing of bovine growth hormone pre-mRNA.

A previous study has demonstrated that deletion of a region within the last exon of bovine growth hormone (bGH) pre-mRNA results in almost complete retention of the upstream intron (Hampson, R. K., LaFollette, L., and Rottman, F. M. (1989) Mol. Cell. Biol. 9, 1604-1610). We now demonstrate that insertion of a simple purine-rich element (GGAAG), which is present within the deleted region, activates intron splicing upon expression in transfected cells. Moreover, several repeats of the GGAA(G) sequence restore splicing to near wild-type levels and direct the binding of a factor present in HeLa cell nuclear extracts. Mutation of the 5'-splice site toward U1 small nuclear RNA complementarity eliminates dependence on the downstream exon sequence for splicing. These results support a model for alternative intron retention in which purine-rich sequences function as part of an "exonic splicing enhancer" to complement a weak 5'-splice site and thereby facilitate intron removal. As a result, the majority of bGH mRNA is processed to remove intron D while still allowing a fraction of bGH mRNA containing the intact intron to reach the cytoplasm.

Alternative Splicing↗

Genetic study of indirect inguinal hernia.

We performed a genetic analysis of 280 families with congenital indirect inguinal hernia ascertained in Shandong province. The multifactorial threshold model and segregation analysis were applied to these families to investigate the mode of inheritance of congenital indirect inguinal hernia. Our results indicate that congenital indirect inguinal hernia is not compatible with a multifactorial threshold model, and the frequent vertical transmission and high segregation ratio suggest autosomal dominant inheritance with incomplete penetrance and sex influence. Through further pedigree analysis of the multiple case families with at least two closely related affected members, we noted preferential paternal transmission of the disease gene, which might suggest the role of genomic imprinting in the aetiology of this condition.

Chi-Square Distribution↗

Expression of HLA-class II genes of IDDM patients on the surface of the LTK- cells.

To investigate the function of HLA-class II genes in the autoimmune response of insulin dependent diabetes mellitus (IDDM), the HLA-class II gene of IDDM patients was introduced into Ltk- cells with pSV2-neo plasmid, using the calcium phosphate precipitation technique. We obtained a stable cell line expressing the HLA-class II gene from lymphocytes of IDDM patients. Expression was identified by direct ox erythrocyte-CrCl3-HLA DR monoclonal antibody rosetting.

Animals↗

[Familial aggregation and HLA typing of pedigrees in insulin-dependent diabetes mellitus].

In 280 patients with IDDM, the positive family history of diabetes was 26.8% of IDDM probands. The prevalence of diabetes in relatives was 68% in first degree relatives, 28% in second degree relatives, and 4% in third degree relatives. HLA typing of 87 members in 13 pedigrees with IDDM was performed. These data support the hypothesis that IDDM is a multigenic hereditary disorder.

Adult↗

In vitro analysis of bovine growth hormone pre-mRNA alternative splicing. Involvement of exon sequences and trans-acting factor(s).

Bovine growth hormone (bGH) pre-mRNA is alternatively spliced, resulting in retention of the last intron (intron D) in a fraction of the cytosolic bGH mRNA. To study the mechanism of this alternative splicing event, we examined the splicing of bGH pre-mRNA in vitro. The splicing of bGH intron D in vitro required a 115-base pair segment of exon 5, reflecting the positive influence of exon sequences observed in transfected cells. No detectable spliceosome complex formation was observed using bGH pre-mRNA containing the 115-base pair deletion in exon 5. The in vitro splicing of the wild type bGH pre-mRNA was inhibited by the addition of RNA containing the 115-nucleotide exon sequence, but not by nonspecific RNAs. UV irradiation of the in vitro splicing reaction resulted in specific cross-linking of a 35-kDa protein(s) to the 115-nucleotide bGH exon sequence. These results suggest that terminal exon sequences are required at an early step of spliceosome complex formation and are consistent with a mechanism in which saturable, trans-acting factor(s) bind to these exon sequences to activate spliceosome complex formation and splicing of bGH intron D.

Alternative Splicing↗

Carbohydrate-binding protein 35. I. Properties of the recombinant polypeptide and the individuality of the domains.

The cDNA clone for carbohydrate-binding protein 35 (CBP35) was engineered into the bacterial expression vector pIN III ompA2, which directs the secretion of the expressed protein into the periplasmic space. Recombinant CBP35 was purified from this system, at a level of approximately 50 mg/liter of bacterial culture. Digestion of recombinant CBP35 with collagenase D, followed by purification using saccharide-specific affinity chromatography yielded a M(r) approximately 16,000 polypeptide, corresponding to the COOH-terminal domain (residues 118-264) of the CBP35 polypeptide. This indicates that the COOH-terminal half of CBP35 contains the carbohydrate recognition domain, consistent with its sequence homology to other S-type lectins. The NH2-terminal domain (residues 1-137) was derived by site-directed mutagenesis of the cDNA, in which stop codons are inserted in place of Gly138 and Gly139, and expression of the mutant cDNA in the same pIN III ompA2 system. The purified NH2-terminal domain failed to bind to saccharide-specific affinity resins. Differential scanning calorimetry of rCBP35 and its individual domains yielded transition temperatures of approximately 39 and approximately 56 degrees C for the NH2- and COOH-terminal domains, respectively. Lactose binding by the COOH-terminal domain shifted the transition temperature to 65 degrees C, whereas sucrose failed to yield the same effect. These results suggest that the individual domains of the CBP35 polypeptide are folded independently.

Amino Acid Sequence↗

In vivo cytokeratin-expression pattern of stratified squamous epithelium from human papillomavirus-type-16-immortalized ectocervical and foreskin keratinocytes.

The association between human papillomavirus (HPV) type 16 and cervical squamous-cell carcinoma has been well documented, and HPV 16 DNA has been shown to immortalize human genital keratinocytes in vitro. Using a panel of cytokeratin(CK)-specific antibodies, we examined the CK expression pattern, an important characteristic of epithelia, of the stratified squamous epithelium reconstructed in vivo from monolayer cultures of 2 human ectocervical and 3 human foreskin keratinocyte cell lines immortalized by HPV 16 DNA. Whereas the abnormal epithelium formed from these grafts presented certain aspects of mature terminal differentiation, such as morphology and expression of CK10/11, the expression patterns for CK19, and especially CK13, were more obviously abnormal. CK18 expression was not detectable in any of the epithelia formed from the 5 cell lines. In contrast, for an HPV-16-immortalized endocervical cell line and the CaSki cervical-carcinoma cell line there was expression of CK18. Our results indicate that HPV-16-induced immortalization of keratinocytes is associated with disruption of the normal CK expression pattern in stratified squamous epithelium and that expression of particular CKs can be differentially disrupted.

Animals↗

Characterization of secretin release in secretin cell-enriched preparation isolated from canine duodenal mucosa.

The release of secretin was studied in secretin cell-enriched preparations isolated from canine duodenal mucosa. The crude enterocytes were isolated by treating the duodenal mucosa sequentially with collagenase and ethylenediaminetetraacetic acid. Secretin cell-enriched fraction was prepared by centrifugation of the crude enterocytes in a counterflow elutriation rotor to obtain a final preparation containing 3.2 +/- 0.3 pmol/10(6) cell of immunoreactive secretin, which was 13-fold greater than the crude cell preparation (N = 5). The cells were incubated in Hanks' balanced salt solution for 20 min at 37 degrees C under 95% O2/5% CO2 before adding various agents and further incubated for various periods of time. The amounts of secretin released into the medium and retained by the cells were then determined by a specific radioimmunoassay. The release of immunoreactive secretin was increased dose-dependently over the control by dibutyryl cyclic-3',5'-adenosine monophosphate, forskolin, 4 beta-12-O-tetradecanoylphorbol-13-acetate, the synthetic serine protease inhibitor, camostat, and the calcium ionophore, A23187. The effects of forskolin, the phorbol ester, and A23187 were time-dependent and not observed at 4 degrees C. The release of immunoreactive secretin was also stimulated by KCl in high concentration and by sodium oleate. The effect of A23187 was abolished in a Ca(2+)-free medium, while those of dibutyryl cyclic-3',5'-adenosine monophosphate and forskolin were potentiated by 3-isobutyl-1-methylxanthine, which did not have a significant effect when added alone. These results indicate that the release of secretin is regulated by both Ca(2+)- and cyclic-3',5'-adenosine monophosphate-dependent mechanisms.2+ release.

Animals↗

General splicing factor SF2/ASF promotes alternative splicing by binding to an exonic splicing enhancer.

The general splicing factor SF2/ASF binds in a sequence-specific manner to a purine-rich exonic splicing enhancer (ESE) in the last exon of bovine growth hormone (bGH) pre-mRNA. More importantly, SF2/ASF stimulates in vitro splicing of bGH intron D through specific interaction with the ESE sequences. However, another general splicing factor, SC35, does not bind the ESE sequences and has no effect on bGH intron D splicing. Thus, one possible function of SF2/ASF in alternative and, perhaps, constitutive pre-mRNA splicing is to recognize ESE sequences. The stimulation of bGH intron D splicing by SF2/ASF is counteracted by the addition of hnRNP A1. The relative levels of SF2/ASF and hnRNP A1 influence the efficiency of bGH intron D splicing in vitro and may be the underlying mechanism of this alternative pre-mRNA processing event in vivo.

Alternative Splicing↗

In vitro and in vivo analysis of cellular origin of cervical squamous metaplasia.

We have previously shown that cultured normal human endocervical cells (HENs) form epithelium resembling squamous metaplasia in vivo. To analyze the cellular origin of squamous metaplasia, the cytokeratin and mucin expression and morphological features of HENs in monolayer cultures and in implants beneath the skin of nude mice were examined. Primary HENs had two distinct morphological phenotypes in vitro pleomorphic epithelial cells and keratinocytelike cells. Using a panel of monoclonal antibodies for various cytokeratins (CKs), we observed that the pleomorphic cells, which were the primary outgrowths, expressed CK7 and CK18 and produced mucin, suggesting their origin to be the mucosecretory columnar cells (CCs) of the endocervix. Keratinocytelike cells were observed in proximity of the CC-like cells after a few days of HEN culture. Interestingly, these cells were homogeneously negative for CK7 expression, as for native reserve cells (RCs), and homogeneously positive for CK13 expression with the antibody that is specific for RCs. During early passages, the culture consisted mostly of the RC-like keratinocytelike cells, and in the late passages, the CC-like cells were predominant. HEN implants in nude mice morphologically formed epithelia similar to immature squamous metaplasia and showed variable CK18 expression. Moreover, they showed homogeneous CK13 expression throughout all layers and expressed mucin and CK7 in the suprabasal cells. The possibility that the HEN culture was originally a mixed population of CCs and RCs, that we failed to detect, cannot be eliminated. Our results support the more likely view that the endocervical simple epithelia, which form squamous metaplasia, are bipotential cells and undergo differentiation readily and reversibly to give rise to CC-like and RC-like cells in culture.

Adult↗

Simple obesity and obesity hyperlipemia treated with otoacupoint pellet pressure and body acupuncture.

161 cases of simple obesity with or without hyperlipemia were treated by pellet pressure on auricular acupoints plus body acupuncture for 3 months in a single-blind experiment. Patients on Capsulae Olei Oenothera Erythrosepalae were taken as positive controls. Body weight, circumference of chest, abdomen, arms and legs, the appetite, sleep, bowel movement, blood TC, TG and HDL-C were recorded for comparison. The results indicated that treatment in the auricular and body acupoint group was superior to treatment in the control group. In the acupoint group body weight dropped by an average of 5.04 kg in 84.55% of the patients, who also showed decreased appetite, blood TC and TG.

Acupuncture Points↗