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Biomedical subjects

G Jiang

Publications and source records attributed to G Jiang.

At least 127 records · Page 7Linked to original sources

[Diagnosis and treatment of pulmonary sequestration].

OBJECTIVE: To explore the cause, classification, incidence, diagnosis and treatment of pulmonary sequestration. METHOD: 42 cases analysed and literatures reviewed. RESULT: 42 cases of sequestration were presents which were resected and confirmed by pathology in our department from 1964 to 1997. 29 cases (69%) were confirmed before operation. The incidence rate was 0.32% comparing with the sametime operations. There were 37 cases (88%) of intralobar type, and 5 cases (12%) of extralobar type. 22 cases accompanied with infection, 9 cases with hemoptysis in intralobar type. 1 case with infection and 1 case with hemoptysis in extralobar type, in the latter case, there was a fistula between the affected lung and the esophagus. CONCLUSION: Pulmonary sequestration is not a rare illness. The diagnosis mainly depends on X-ray. Sometimes, bronchography, retrograde arteriography or CT can also make the diagnosis correctly. Special attention should be paid to avoiding injure to the vessels which supply the isolated lung during operation.

Adolescent↗

[Reliability design and analysis of man-machine performance of display and illumination system in cockpit].

This paper analysed and discussed the basic reliability of display and illumination system in which Chinese characters displayed on LCD were read and the colours were identified, and instruments in different visual views and the instruments of differential display models were read. The colour rendering of the light source was also evaluated. Basic reliability may be chosen as 0.9995.

Aircraft↗

[Lung autotransplantation technique in the treatment of stage III bronchogenic carcinoma].

OBJECTIVE: To perform a double sleeve right upper and middle bilobectomy with transposing the inferior pulmonary vein to the superior one stump in a lung cancer patient. METHOD: The "cut-down" lobe was immersed in heparin solution in vitro for 15 minutes. It took 15, 14 and 10 minutes to finish the anastomosis of pulmonary vein, bronchus, and artery. The pulmonary artery was blocked for 3 hours. Postoperatively, the patient underwent ventilation for 6 hours. RESULT: In the 5th day after operation, re-implanted lung inflated well, the chest tube was retreated, and the patient can walk around. The reconstruction of pulmonary vessels and airway was done by spiral CT, which demonstrated no stenoses of bronchi and the pulmonary blood perfused and returned very well 3 weeks after operation. Follow-up more than 6 months showed the patient had a good quality of life and resumed his work. CONCLUSION: It is indicated to apply the technique of lung autotransplantation for operation on the central bronchogenic carcinoma when the main bronchus or pulmonary artery involved by tumor is too long to perform the anastomosis in the double sleeve bilobectomy.

Bronchi↗

Betidamino acid scan of the GnRH antagonist acyline.

Strong clinical evidence suggests that GnRH antagonists will replace GnRH agonists in a number of indications because of their ability to inhibit gonadotropin secretion as long as an adequate concentration of the analogue is present in the circulation whereas superagonists will take approximately 2 weeks to desensitize the gonadotrophs. Until recently, antagonists were either too weak and/or would release histamine. Azaline B {[Ac-D2Nal1,D4Cpa2,D3Pal3, 4Aph5(atz),D4Aph6(atz),ILys8,DAla10] GnRH} and long-acting members of the azaline family {Ac-D2Nal-D4Cpa-D3Pal-Ser-4Aph(X)-D4Aph(Y) -Leu-ILys-Pro-DAla-NH2}, however, appear to be promising drug candidates. Because these antagonists tend to form gels (due to the formation of beta-sheet structures) and, as a result, are not readily amenable to formulation for long-term delivery, we have investigated ways of increasing hydrophilicity while retaining high potency and lack of histamine releasing activity. Betidamino acids (a contraction of "beta" position and "amide") are N'-monoacylated (optionally, N'-monoacylated and N-mono- or N,N'-dialkylated) aminoglycine derivatives in which each N'-acyl/alkyl group may mimic naturally occurring amino acid side chains or introduce novel functionalities. We have used unresolved N alpha-Boc,N'alpha-Fmoc-aminoglycine, and N alpha-Boc,N'alpha-(CH3)Fmoc-aminoglycine as templates for the introduction of betidamino acids in acyline (Ac-D2Nal-D4Cpa-D3Pal-Ser-4Aph(Ac)-D4Aph(A c)-Leu-Ilys-Pro-DAla-NH2), a long acting member of the azaline B family, to test biocompatibility of these betide derivatives. Diastereomeric peptides could be separated using RP-HPLC in most cases. Biological results obtained in vitro (binding affinity to rat pituitary gland membranes) and in vivo (rat antiovulatory assay, AOA) indicate in most cases small differences in relative potencies (< 5-fold) between the D- and L-nonalkylated betidamino acid-containing acylines. Importantly, most betide diastereomers have high affinity for the GnRH receptor and were equipotent with acyline in the AOA. Greater differences in affinity and potency between diastereomers were observed after introduction of a methyl group on the side chain nitrogen ("beta" position) of the same analogues, with one of the diastereomer having an affinity and a potency in the AOA equivalent to that of acyline. These results suggest that chirality at the alpha-carbon coupled to side chain orientation is important for receptor recognition. The duration of action of some of the most potent analogues was also determined in the castrated male rat in order to measure the extent (efficacy and duration of action) of inhibition of luteinizing hormone release. Data suggest that introduction of a betidamino acid results in reduction of duration of action. Also, introduction of betidamino acids results in peptides with increased hydrophilicity (as determined by elution times on C18 silicas at pH 7.3) compared to that of the parent compound. N'-Methyl substitution results in parallel increase in retention times on C18 silicas as expected.

Amides↗

Serine/threonine phosphorylation of orphan receptor hepatocyte nuclear factor 4.

We showed previously that hepatocyte nuclear factor 4 (HNF-4) defines a new subclass, Group IV, of nuclear receptors. In order to determine whether members of this subclass are phosphorylated, HNF-4 was overexpressed to high levels in insect cells using a baculovirus expression system. The baculovirus-expressed HNF-4 (HNF4.BV) was characterized and compared to HNF-4 overexpressed in transiently transfected mammalian (COS-7) cells (HNF4.COS). The results indicate that both HNF4.BV and HNF4.COS are phosphorylated although HNF4.BV was hypophosphorylated relative to HNF4.COS. Phosphoamino acid analysis showed that HNF-4 is phosphorylated mainly on serine and to a lesser extent on threonine residues. Phosphopeptide mapping revealed 13 phosphopeptides for HNF4.COS, only 9 of which were present in the HNF4.BV sample. DNA-binding studies also showed that HNF4.BV binds DNA with a lower specificity and affinity, as measured by the equilibrium dissociation constant (Kd), than does HNF4.COS. Partial proteolytic digestion experiments also revealed that HNF4.BV and HNF4.COS adopt somewhat different three-dimensional conformations. Since glycosylation of HNF4.BV was ruled out by a number of methods and since HNF-4 expressed in bacteria exhibited an even lower DNA-binding affinity than HNF4.BV, we propose that serine/theronine phosphorylation may play a role in the DNA-binding activity of HNF-4 and, therefore, possibly of other Group IV receptors as well.

Animals↗

The DNA binding domain of hepatocyte nuclear factor 4 mediates cooperative, specific binding to DNA and heterodimerization with the retinoid X receptor alpha.

We recently showed that hepatocyte nuclear factor 4 (HNF-4) defines a unique subclass of nuclear receptors that exist in solution and bind DNA elements as homodimers (Jiang, G., Nepomuceno, L., Hopkins, K., and Sladek, F. M. (1995) Mol. Cell. Biol. 15, 5131-5143). In this study, we show that the dimerization domains of HNF-4 map to both the DNA binding and the ligand binding domain. Whereas the latter is critical for dimerization in solution, the DNA binding domain mediates cooperative, specific binding to direct repeats of AGGTCA separated by one or two nucleotides. Whereas amino acid residues 117-125 (the T-box/third helix region) are insufficient for cooperative homodimerization and high affinity DNA binding, residues 126-142 (encompassing the A-box region) are required. Finally, in contrast to the full-length receptor, the DNA binding domain of HNF-4 is capable of heterodimerizing with that of the retinoid X receptor alpha but not with that of other receptors. These results indicate that the HNF-4 DNA binding domain is distinct from that of other receptors and that the determinants that prevent HNF-4 from heterodimerizing with RXR lie outside the DNA binding domain, presumably in the ligand binding domain.

Amino Acid Sequence↗

PCR detection of human papillomavirus: comparison between MY09/MY11 and GP5+/GP6+ primer systems.

Human papillomavirus (HPV) is an etiologic agent of cervical cancer and is the most common sexually transmitted disease in women. PCR amplification of HPV genomes is the most sensitive method for the detection of cervicovaginal HPV. We have compared the two most commonly used PCR primer sets, MY09/MY11 (MY-PCR) and GP5+/GP6+ (GP+-PCR), for the detection of HPV DNA in cervicovaginal lavage samples from 208 women. Oligonucleotide probes for 39 different HPV types were used. Both primer sets amplified a wide spectrum of HPV genotypes and detected similar overall prevalences of 45% (94 of 208) and 43% (89 of 208), respectively. The MY-PCR system detected 27 of 30 (90%) samples with multiple HPV types, whereas the GP+-PCR system detected 14 of 30 (47%) samples with multiple HPV types. Differences in the detection of HPV types 35, 53, and 61 were noted between the two primer systems. Serial dilution of plasmid templates indicated a 3-log decrease in the amplification of HPV type 35 by MY-PCR and HPV types 53 and 61 by GP+-PCR. These results indicate that although the MY-PCR and GP+-PCR identified nearly equivalent prevalences of HPV in a set of clinical samples, differences in the detection of specific types and infections with multiple types were found. Differences in the sensitivities and characteristics of the PCR systems for the detection of HPV within clinical samples should be considered when comparing data between studies and/or in designing new studies or clinical trials.

DNA Primers↗

Proposed mechanism for the stabilization of nuclear receptor DNA binding via protein dimerization.

Hepatocyte nuclear factor 4 (HNF-4) defines a new subgroup of nuclear receptors that exist in solution and bind DNA exclusively as homodimers. We recently showed that the putative ligand binding domain (LBD) of HNF-4 is responsible for dimerization in solution and prevents heterodimerization with other receptors. In this report, the role of the LBD in DNA binding by HNF-4 is further investigated by using electrophoretic mobility shift analysis. A comparison of constructs containing either the DNA binding domain (DBD) alone or the DBD plus the LBD of HNF-4 showed that dimerization via the DBD was sufficient to provide nearly the full DNA binding affinity of the full-length HNF-4. In contrast, dimerization via the DBD was not sufficient to produce a stable protein-DNA complex, whereas dimerization via the LBD increased the half-life of the complex by at least 100-fold. Circular permutation analysis showed that full-length HNF-4 bent DNA by approximately 80 degrees while the DBD bent DNA by only 24 degrees. Nonetheless, analysis of other constructs indicated that the increase in stability afforded by the LBD could be explained only partially by an increased ability to bend DNA. Coimmunoprecipitation studies, on the other hand, showed that dimerization via the LBD produced a protein-protein complex that was much more stable than the corresponding protein-DNA complex. These results led us to propose a model in which dimerization via the LBD stabilizes the receptor on DNA by converting an energetically favorable two-step dissociation event into an energetically unfavorable single-step event. Implications of this one-step model for other nuclear receptors are discussed.

Binding Sites↗

[Measurements of serum and subretinal fluid before and after surgery of retinal detachment].

OBJECTIVES: To investigate the relationships between proliferative vitreoretinopathy (PVR) formation and the levels of human epidermal growth factor (h-EGF) and tumor necrosis factor (TNF) in serum and subretinal fluid (SRF) before and after surgery of retinal detachment with PVR. METHODS: Serum and SRF samples of 51 patients with retinal detachment and PVR were collected, and their h-EGF and TNF levels were measured by radioimmunoassays. RESULTS: In comparison with the control group, the levels of h-EGF and TNF in serum and SRF in retinal detachment with PVR group were higher but not statistically significant (P > 0.05). Their levels in the serum were not related to the severity of the disease, while their levels in SRF increased significantly with the increase of the severity of the disease, and these two factors were positively correlated (P < 0.05). 5 days after the surgery of retinal detachment, the h-EGF and TNF levels in serum increased significantly (P < 0.01) and the levels decreased within 30 days but not to normal. CONCLUSIONS: The h-EGF and TNF in SRF might have a synergic action and affect the formation and development of PVR. The h-EGF and TNF in serum take part in the repair of the operative trauma and inflammatory reaction. However, their high levels could interfere with the effect of operation.

Adult↗

[Expression of hepatitis B virus S genes in Ad5 vector].

The hepatitis B virus (HBV) S gene, PreS2 + S genes and the late phase expression cassette (MTI) + HBV S genes were separately cloned into Ad5 vector downstream of E3 promoter (pAd5 deltaE3 provided by Wyeth Co.). The above constructed plasmids and Ad5 DNA EcoR I A fragment were cotransfected into 293 cells. The progeny adenoviruses named rAd5S, rAd5MS, rAd5S2S were harvested for analysis. The recombinants were isolated and analyzed by PCR, using two primers specific to the HBV S genes. The expressed products were detected by ELISA and RIA. The recombinant containing MIT + HBV S genes (rAd5MS) was identified to be ELISA positive, whereas the other two recombinants (rAd5S, rAd5S2S) were negative to ELISA, but positive to RIA. The results indicated that adenovirus E3 early promoter could express the inserted foreign genes, and MIT worked well in the E3 region of Ad5 and could increase the expression capacity of the recombinants. The conditions for foreign gene expression and genetic stability of the recombinant viruses were studied in detail. There was no wild Ad5 discovered during the cotransfection experiments. The present study provides some experiences for studying adenovirus recombination.

Adenoviridae↗

Betidamino acids: versatile and constrained scaffolds for drug discovery.

Betidamino acids (a contraction of "beta" position and "amide") are N'-monoacylated (optionally, N'-monoacylated and N-mono- or N,N'-dialkylated) aminoglycine derivatives in which each N'acyl/alkyl group may mimic naturally occurring amino acid side chains or introduce novel functionalities. Betidamino acids are most conveniently generated on solid supports used for the synthesis of peptides by selective acylation of one of the two amino functions of orthogonally protected aminoglycine(s) to generate the side chain either prior to or after the elongation of the main chain. We have used unresolved Nalpha-tert-butyloxycarbonyl-N'alpha-fluorenylmethoxycarbonyl++ + aminoglycine, and Nalpha-(Nalpha-methyl)-tert-butyloxycarbonyl-N'alpha-fluo renylmethoxycarbonyl aminoglycine as the templates for the introduction of betidamino acids in Acyline [Ac-D2Nal-D4Cpa-D3Pal-Ser-4Aph(Ac)-D4Aph(A c)-Leu-Ilys-Pro-DAla-NH2, where 2Nal is 2-naphthylalanine, 4Cpa is 4-chlorophenylalanine, 3Pal is 3-pyridylalanine, Aph is 4-aminophenylalanine, and Ilys is Nepsilon-isopropyllysine], a potent gonadotropin-releasing hormone antagonist, in order to test biocompatibility of these derivatives. Diasteremneric peptides could be separated in most cases by reverse-phase HPLC. Biological results indicated small differences in relative potencies (<5-fold) between the D and L nonalkylated betidamino acid-containing Acyline derivatives. Importantly, most betide diastereomers were equipotent with Acyline. In an attempt to correlate structure and observed potency, Ramachandran-type plots were calculated for a series of betidamino acids and their methylated homologs. According to these calculations, betidamino acids have access to a more limited and distinct number of conformational states (including those associated with alpha-helices, beta-sheets, or turn structures), with deeper minima than those observed for natural amino acids.

Amino Acid Sequence↗

[A genetic epidemiological study on lung cancer].

Segregation ratio, heritability and relative risk of genetic susceptibility were estimated for 355 families of lung cancer in matched pair with genetic epidemiological methods. Results showed that segregation ratio for lung cancer was 0.09-0.12 (95% confidence interval, CI) and heritability of lung cancer was (40.58 +/- 4.01)% and (27.58 +/- 4.76)% for smokers and non-smokers, respectively. Relative risks of genetic susceptibility to lung cancer were 4.73 (95% CI 3.90-5.74) and 2.61 (95% CI 2.18-3.13) in their first and second degree relatives, respectively, after adjustment of smoking with a logistic regression model. It was also found that there was an interaction between smoking habit and genetic background of lung cancer. Thus, it is believed that genetic background is one of the multifactorial causes in lung cancer.

China↗

[Assay of contents of lysozyme, SIgA, IgA and IgG in tongue coating].

The contents of lysozyme, SIgA, IgA, and IgG in 98 cases with pathological tongue coating and 30 cases of normal coating were analysed by using the thorough random analysis of variance. The results showed that the content of lysozyme in pathological coating was lower than that in normals significantly (P < 0.0001), while the contents of SIgA, IgG in pathological coating were higher than that of normals (P < 0.0001). It indicates that immune reaction is very active during the development of pathological tongue coating.

Adolescent↗

Molecular cloning and functional analysis of the promoter of the human squalene synthase gene.

We have cloned and characterized the 5'-flanking region of the gene encoding human squalene synthase. We report here the promoter activity of successively 5'-truncated sections of a 1 kilobase of this region by fusing it to the coding region of a luciferase reporter gene. DNA segments of 200 base pairs (bp) 5' to the transcription start site, as determined by primer extension analysis, show a strong promoter effect on the expression of the luciferase chimeric gene and a high response to the presence of sterols when transiently transfected into the human hepatoma cell line HepG2 or to the hamster-derived CHO-K1 cells. An approximately 50-fold induction of luciferase activity, in the absence of sterols, was observed in transiently transfected HepG2 cells for fusion constructs containing sections of 200, 459, and 934 bp of the putative human squalene synthase promoter. Loss of promoter activity and response to sterols was localized to a 69-bp section located 131 nucleotides 5' to the transcription start site. Sequence analysis of this region showed that it contained a sterol regulatory element 1 (SRE-1) previously identified in other sterol regulated genes (Smith, J. R., Osborne, T. F., Brown, M. S., Goldstein, J. L., and Gil, G. (1988). J. Biol. Chem. 263, 18480-18487) and two potential NF-1 binding sites. Additional CCAAT box, SRE-1 element, and two Sp1 sites were identified 3' to this section. Sequences within this 69-bp DNA, including the SRE-1 cis-acting element, show strong binding to the purified nuclear transcription factor ADD1 (Tonzonoz, P., Kim, J. B., Graves, R. A., and Spiegelman B. M. (1993) Mol. Cell Biol. 13, 4753-4759) by mobility shift assay and footprinting analyses.

Base Sequence↗

Gonadotropin-releasing hormone antagonists: novel members of the azaline B family.

A series of antagonists of gonadotropin-releasing hormone (GnRH) homologous to azaline B ([Ac-DNal1,DCpa2,DPal3,Aph5(Atz),DAph6+ ++(Atz),ILys8,DAla10]GnRH) was synthesized, characterized, and tested in a rat antiovulatory assay (AOA). Selected analogues were also tested in both an in vitro dispersed rat pituitary cell culture assay for inhibition of GnRH-stimulated luteinizing hormone release and an in vitro histamine release assay. The duration of action of some of the most potent and safest analogues in those assays was also determined in the castrated male rat in order to measure the extent (efficacy and duration of action) of inhibition of luteinizing hormone release. Structurally, this series of analogues has novel substitutions (X and Y) in the structure of the azaline B precursor: [Ac-DNal1,DCpa2,DPal3,-Aph5(X),DAph6(Y),++ +ILys8,DAla10]GnRH. These substitutions were designed to confer increased hydrophilicity as compared to that of azaline B (determined by relative retention times on a C18 reverse phase column using a triethylammonium phosphate buffer at pH 7.3) or to make them more easily accessible synthetically. Some bulky substituents were introduced in order to probe the spatial limitations of the receptor's cavity. These substitutions include acylated 4-aminophenylalanine at positions 5 and/or 6 (29 analogues), N alpha-methylated backbone substitutions (six analogues), N omega-isopropylaminophenylalanine at position 8, and hydrophilic amino acids at position 1. Out of 20 novel analogues tested for long duration of action in this series, only seven ([Ac-DNal1,DCpa2,DPal3,Aph5,DAph6,ILys8 ,DAla10]GnRH, [Ac-DNal1,DCpa2,DPal3,Aph5(For),DAph6(For) ,ILys8,DAla10]GnRH, [Ac-DNal1,DCpa2,DPal3,Aph5(Ac),DAph6(Ac),- ILys8,DAla10]GnRH (acyline), [Ac-DNal1,DCpa2,DPal3,Aph5(Pio),DAph6++ +(Pio),ILys8,DAla10]GnRH, [Ac-DNal1,DCpa2,DPal3,Aph5(Atz),DAph6++ +(Ac),ILys8,DAla10]GnRH, [Ac-DNalDCpa2,DPal3,Aph5(Atz-beta Ala),DAph6(Atz-beta Ala),ILys8, DAla10]GnRH, [Ac-DNal1,DCpa2,DPal3,Aph5(Atz-Gab), DAph6(Atz-Gab),ILys8,DAla10]GnRH) had relative potencies and/or duration of action comparable to those of azaline B. The others were one-half to one-tenth as effective as azaline B. N alpha-Methylated backbone substitutions at position 5 yielded analogues that were significantly more hydrophilic presumably because of the breakage of the NH alpha-Tyr5 to Arg8-CO hydrogen bond reported to stabilize a beta-turn encompassing residues 5-8 and which favored beta-sheet formation as shown earlier by Haviv et al. This substitution resulted, however, in an increased potency in the histamine release assay and in significantly shorter duration of action. Similarly, attempts at replacing isopropyllysine in position 8 by either isopropyl-4-aminophenylalanine or isopropyl-4-(aminomethyl)phenylalanine resulted in loss of potency in the AOA. Changes in chirality at position 1 or 10 resulted in analogues that were one-tenth and one-half as potent, respectively, as acyline.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗

Genetic characterization of the human papillomavirus (HPV) 18 E2 gene in clinical specimens suggests the presence of a subtype with decreased oncogenic potential.

HPV 18 is associated with 2 divergent phenotypes: (i) aggressive cervical cancer and a preponderance of cancer relative to cervical intra-epithelial neoplasia (CIN) and (ii) benign warty lesions of the cervix. The E2 gene of HPV 18 encodes a regulatory protein that represses viral oncogene transcription and is involved in viral replication. Variation within the E2 gene of HPV 18 and its correlation with the morphologic grade of associated lesions were analyzed in a sample of 20 HPV 18-positive cervical specimens representing a spectrum of pathology from low-grade CIN to cervical cancer. An amplification HPV 18 E2 gene was present in 3 of 5 cancers, indicating that E2 disruption was not required for cancer development. Single-strand conformation polymorphism and PCR analyses revealed a high degree of polymorphism throughout the E2 gene. Direct DNA sequencing of both strands of a 154-bp fragment revealed a variability of 5.8%. Six intra-epithelial lesions contained alterations in common that account for 3.9% of the variation and appear to constitute a subtype. Within the 154-bp region, 2 of 3 cervical cancers and 0 of 12 intra-epithelial lesions were identical to the published HPV 18 sequence. DNA sequence analysis of a region extending into the E5 open reading frame revealed deletions in the E2/E5 intragenic region that were present in 50% of the members of the subtype. Our data demonstrate significant sequence variation within the E2 gene and suggest the presence of an HPV 18 subtype with decreased oncogenic potential.

Base Sequence↗