Biomedical subjects
Publications and source records attributed to X Zeng.
In this paper, pharmacognostiacal identification of Humulus scandens was studied. The character of medicinal materials, histological and powder characteristics for the setem and leaf of H. scandens were mainly reported. The microscopical features for leaf is typical identified basis.
The dependence of net analyte emission intensity and signal-background ratio on coolant gas composition and observation heights for 8 lines of four elements (Mg, Cr, Cd and Mn) in Ar-air mixed coolant gas ICP was measured under the similar condition as that in the low power Ar-ICP with a commercial sequential plasma spectrometer. Further, the detection limits under compromise conditions were estimated. The results show that ion lines and neutral atom lines with high excitation energy have maximum intensity and are more intense than those in the Ar-ICP, when 5-10% air is added to the coolant flow. The intensity of neutral atom lines with low excitation energy drops gradually as air is added to coolant flow. The signal-background ratio of analyte lines depends upon excitation energy and wavelength.
OBJECTIVE: This article reports that competitive hybridization using entire chromosome specific libraries as probe and human genomic DNA as the competitor allows intense and specific fluorescent staining of human chromosome in metaphase. This general approach is called "chromosome painting". METHODS: The probes comprising chromosomes 2, 5, 6, 7, 13, 14, X specific libraries were used to analyse five cases which had been suspected of subtle translocation and deletion in karyotype analysis by G-banding of metaphase cells. The authors selected entire chromosome-specific DNA libraries hybridizing with the five cases. Unlabeled human genomic DNA was used to inhibit the hybridization of sequences in the library that bind to multiple chromosome. RESULTS: The target chromosome was made at least 20 times brighter parunit length than the others. Translocations and deletions were detected clearly in metaphase and were consistent with G-banding. However, the result was clearer and the detection easier, compared with G-banding. CONCLUSION: Chromosome painting is very powerful for identification of chromosome structural aberrations. Translocation and deletion involving these chromosomes can be strikingly visualized. The hybridization intensity and specificity are such that even very small portions of the involved chromosome can be detected. This technique is especially useful in settings where high-quality banding is difficult.
Thyroid transcription factor-1 (TITF-1) is a homeodomain containing transcription factor that binds to and selectively activates the expression of genes in thyroid and pulmonary epithelial cells. TITF-1 plays a critical role in gene expression and in organogenesis of lung and thyroid. In the present work, epitope-tagged TITF-1 proteins were used to identify the regions of the TITF-1 polypeptide that mediate nuclear localization and transcriptional activity in human lung adenocarcinoma cells. A series of TITF-1-flag deletion mutants was generated and transfected into H441 cells to determine amino acid sequences involved in translocation to the nucleus. Transfection of the TITF-1-flag mutants demonstrated that a nuclear localization signal (NLS) sequence, located at the N-terminus of the homeodomain, is critical for nuclear targeting. The NLS was essential but not sufficient for translocation of TITF-1 to the nucleus, since deletion of the homeodomain itself also blocked nuclear translocation in the presence of NLS. Deletion of the N-terminal transactivation domain of TITF-1 completely abolished its transcriptional activation on the human surfactant protein-B promoter, and deletion of the C-terminal domain partially reduced its stimulatory activity. Nuclear translocation of TITF-1 depends on both an NLS and the homeodomain of the polypeptide. Both C- and N-terminal regions of TITF-1 are involved in transactivation of surfactant protein B gene expression in pulmonary cells.
Regulation of gene expression by many transcription factors is controlled by specific combinations of homo- and heterodimers through a short alpha-helical coiled-coil known as a leucine zipper. The dimer interface of a leucine zipper involves side chains of the residues at the a, d, e, and g positions of the (abcdefg)n heptad repeat. To understand the basis for the specificity of dimer formation, we characterized GCN4 leucine zipper mutants with all 16 possible permutations and combinations of isoleucines and asparagines at four a positions in the dimer interface, using a genetic test for the specificity of dimer formation by lambda repressor-leucine zipper fusions. Heterodimers were detected by loss of repressor activity in the presence of a fusion to a dominant-negative mutant form of the DNA-binding domain of repressor. Reconstruction experiments using leucine zippers from GCN4, Jun, Fos, and C/EBP showed that this assay distinguishes pairs that form heterodimers from those that do not. We found that the mutants have novel dimerization specificities determined by the positioning of buried asparagine residues at the a positions. The pattern of buried polar residues could also explain the dimerization specificities of some naturally occurring leucine zippers. The altered specificity mutants described here should be useful for the construction of artificial regulatory circuitry.
Chimeric proteins comprising the N-terminal DNA binding domain of lambda repressor fused to a fragment of a foreign protein have been used to detect oligomerization of the latter. Fusions containing dimeric and tetrameric leucine zipper domains can be distinguished based on their in vivo repressor activities on a pair of cat-lacZ reporter strains. Repressor fusions are unable to efficiently repress transcription from a synthetic promoter that overlaps a weak operator site; repression by tetrameric, but not dimeric, fusion proteins is increased by the presence of a strong, upstream operator site. To construct reporters we developed a shuttle system that allows rapid construction of single-copy operon fusions in E. coli, with both cat and lacZ as reporters.
Nasopharyngeal carcinoma (NPC) is rare in most parts of the world but occurs with high incidence in certain regions, such as South-East Asia. Two major histological types of NPC are recognized, non-keratinizing carcinoma and squamous cell carcinoma. Non-keratinizing NPCs, which include undifferentiated NPC, are invariably associated with Epstein-Barr virus (EBV) infection, regardless of the geographical or ethnic origin of the patients. By contrast, conflicting results have been published concerning a possible association of squamous cell NPC with the virus. To address this question, squamous cell NPCs have been collated from an area where NPC is endemic, Hong Kong, and from two regions where NPC occurs with a lower incidence, Chengdu, PR China, and Birmingham, United Kingdom. In situ hybridization for the detection of the small EBV-encoded nuclear RNAs (EBERs) demonstrated that all 22 cases from Hong Kong were EBV-positive. By contrast, EBV was detectable in 7 of 19 cases from central China, and in 3 of 7 cases from the U.K. Expression of the virus-encoded latent membrane protein 1 (LMP1) was detected in 3 of 32 EBV-positive squamous cell NPCs. These results indicate that the association of squamous cell NPCs with EBV shows geographical variability in a manner which is reminiscent of the situation encountered in Burkitt's lymphoma. This suggests that squamous cell NPCs are a pathogenetically heterogeneous group of tumours distinct from non-keratinizing NPCs.
GABAergic synaptic responses were studied by direct, monosynaptic activation of GABAergic interneurons in the CA1 region of in vitro hippocampal slices from rats made tolerant to the benzodiazepine, flurazepam. Monosynaptic IPSPs were elicited in CA1 pyramidal neurons, following 1 week oral flurazepam administration, by electrical stimulation at the stratum oriens/stratum pyramidale or stratum radiatum/ stratum-lacanosum border < or = 0.5 mm from the recording electrode plane. Excitatory input to pyramidal cells and interneurons was eliminated by prior superfusion of the glutamate receptor antagonists, APV (50 microM) and DNQX (10 microM). GABAA receptor-mediated early IPSPs were further isolated by perfusion of the GABAB antagonist, CGP 35348 (25 microM) or by diffusion of Cs- from the recording electrode. GABAB receptor-mediated late IPSPs were pharmacologically isolated by perfusion of the GABAA antagonist, picrotoxin (50 microM). There was a significant decrease in the amplitude of pharmacologically isolated early and late IPSPs in FZP-treated neurons without a change in passive membrane properties. A shift of the early IPSP, but not the late IPSP, reversal potential in FZP-treated neurons suggested that a change in the driving force for anions, presumably Cl, in CA1 neurons was one important factor related to the decreased early IPSP amplitude after prolonged activation of GABAA receptors by flurazepam. A decreased early IPSP amplitude accompanied by a decreased late IPSP amplitude suggested that presynaptic GABA release onto FZP-treated pyramidal cells may also be reduced. We conclude from these data that an impairment of GABAergic transmission in CA1 pyramidal neurons associated with the development of tolerance during chronic benzodiazepine treatment may be related to the regulation of both pre- and postsynaptic mechanisms at the GABA synapse.
Putative intersubunit electrostatic interactions between charged amino acids on the surfaces of the dimer interfaces of leucine zippers (g-e' ion pairs) have been implicated as determinants of dimerization specificity. To evaluate the importance of these ionic interactions in determining the specificity of dimer formation, we constructed a pool of > 65,000 GCN4 leucine zipper mutants in which all the e and g positions are occupied by different combinations of alanine, glutamic acid, lysine, or threonine. The oligomerization properties of these mutants were evaluated based on the phenotypes of cells expressing lambda repressor-leucine zipper fusion proteins. About 90% of the mutants do not form stable homooligomers. Surprisingly, approximately 8% of the mutant sequences have phenotypes consistent with the formation of higher-order (> dimer) oligomers, which can be classified into three types based on sequence features. The oligomerization states of mutants from two of these types were determined by characterizing purified fusion proteins. The Type I mutant behaved as a tetramer under all tested conditions, whereas the Type III mutant formed a variety of higher-order oligomers, depending on the solution conditions. Stable homodimers comprise less than 3% of the pool; several g-e' positions in these mutants could form attractive ion pairs. Putative repulsive ion pairs are not found among the homodimeric mutants. However, patterns of charged residues at the e and g positions do not seem to be sufficient to predict either homodimer or heterodimer formation among the mutants.
The multifunctional regulatory factor Rap1p of Saccharomyces cerevisiae accomplishes one of its tasks, transcriptional activation, by complexing with Gcr1p. An unusual feature of this heteromeric complex is its apparent capacity to contact simultaneously two adjacent DNA elements (UASRPG and the CT box, bound specifically by Rap1p and Gcr1p, respectively). The complex can activate transcription through isolated UASRPG but not CT elements. In promoters that contain both DNA signals its activity is enhanced, provided the helical spacing between the two elements is appropriate; this suggests that at least transient DNA loop formation is involved. We show here that this CT box-dependent augmentation of Rap1p/Gcr1p activation requires the presence of a third protein Gcr2p; the Gcr2- growth defect appears to result from a genome-wide loss of the CT box effect. Interestingly, a hyperphosphorylated form of Gcr1p disappears in delta gcr2 cells but reappears if they harbor a doubly point-mutated GCR1 allele that bypasses the Gcr2- growth defect. Gcr2p therefore appears to induce a conformation change in Gcr1p and/or stimulate its hyperphosphorylation; one or both of these effects can be mimicked in the absence of GCR2 by mutation of GCR1. This improved view of Rap1p/Gcr1p/Gcr2p function reveals a new aspect of eukaryotic gene regulation: modification of an upstream activator, accompanied by at least transient DNA loop formation, mediates its improved capacity to activate transcription.
The cellular interleukin-6 (IL-6) gene contains a target site for the mammalian transcriptional repressor RBP. The target site is contained within the interleukin response element (ILRE), which mediates IL-6 activation by NF-kappa B. In this study, we show by using transient-expression assays that RBP represses activated transcription from the IL-6 gene. The presence and position of the RBP target site are crucial in mediating repression by RBP. While RBP binds within the ILRE, it does not target NF-kappa B alone; nonetheless, NF-kappa B binding to the ILRE is required for repression. Our results indicate that RBP represses coactivation by NF-kappa B and another cellular transcription factor, C/EBP-beta.
Cross reactions of T. spiralis muscular larva antigens (TsMLA) against several sera from patients with other parasitic diseases by ELIB (enzyme-linked immunoblotting technique) were studied. The proteins of 31-100 KDa were recognized by sera of two cases of acute schistosomiasis, while proteins of 44/45, 51/53, 62/64 and 100 KDa were recognized by sera of 11 cases of chronic schistosomiasis. The proteins of 60 KDa showed varied cross reactions against sera of five kinds of patients with other parasitic diseases (filariasis, hookworm diseases, paragonimiasis, ascariasis and clonorchiasis). Only the proteins of 45 KDa were recognized by six of fifteen cases of normal human sera. All of > 29 KDa proteins were recognized by sera of rabbit infected by T. spiralis. It is suggested that there were many antigens in Trichinellen muscular larva which shared in common with several other parasites. The cross reactions of TsMLA against Schistosoma japonicum may be differentiated on the positive reaction bands of 44/45, 51/53, 62/64 and 100 KDa proteins which was specific to the chronic schistosomiasis using the TsMLA by means of ELIB.
In order to identify antigens in circulating immune complexes (CIC) of Schistosoma japonicum, the antiserum against CIC was first successfully prepared by immunization of New Zealand rabbit with CIC isolated from schistosome-infected rabbit sera. The antigens in CIC were identified and analyzed by EITB (enzyme-linked immunoelectro-transfer blot) techniques, using the prepared antiserum against CIC, adult worm antigens (AWA), soluble egg antigens (SEA) and adult excretory antigens (ESA). The results showed that the antiserum against CIC had recognized the AWA bands of 155, 90, 70, 31/32, 20.5, 14 and 8 KD; the SEA bands of 90, 66, 46, 34.5 and 27 KD and the ESA bands of 155, 92, 87.5, 70, 60, 57.5, 50, 46, 36, 31/32, 24, 18.5, 12 and 8 KD. These findings indicate that the antigens in CIC were derived from both schistosome adult worms and eggs. In addition, the localization of the antiserum against CIC targeted antigens in adult worms was also carried out by IFAT (indirect flourescent antibody test). It is suggested that the antigens in CIC were presented mainly in the tegument and the gut of the parasite.
The subclass antibodies against IgG1, IgG3 and IgG4 in sera of the patients with chronic Schistosomiasis japonica were detected before treatment, and after treatment--6 and 12 months respectively, using Biotin-Avidin-ELISA (BA-ELISA) established by purified 31/32 KD antigen from the adult worms. At the same time IgG1 and IgM were examined by the standard ELISA. False positive reaction with normal control and cross reaction with other parasitic diseases have not been observed. The IgG1 and IgG3 subclasses showed high sensitivity and specificity and reduced quickly 6 months after treatment. These results indicate that the level of specific IgG1 and IgG4 to the 31/32 KD adult worm protein has high value for diagnosis of Schistosomiasis japonica and evaluation of the curative efficiency of the disease.
The effects of different components of serum from radiation, burn and combined radiation-burn injury on the activity of L-type calcium channel in cultured myocardial cells were investigated in this study. The results revealed that the serum from different injury groups could all activate the L-type calcium channel of cultured myocardial cells. These changes altered the calcium level in cells, which might be an important cause of dysfunction in cardiovascular system. Among the components of postinjury serum, the effect of high molecular components (> 8-10 kD) was not significant, whereas low molecule (< 8-10kD) and lipid components were the major effective components. The low molecule of postinjury serum affected not only on the opening activity of L-type channel, but also on the state of molecular movement in the membrane. The effect of serum lipids from injury groups could be depressed by SOD, which suggests that there may be free radical reactions during the serum lipids effect on calcium channel. The effects of serum and its components from combined radiation-burn group were the most intensive, and those from radiation injury were the weakest. The nature of the toxic material in serum remains to be further investigated.
OBJECTIVE: To construct and identify Escherchia coli (E. coli)-Mycobacteria shuttle plasmid and to detect stable expression of foreign gene in E. coli and BCG. METHOD: With a genetic engineering technique to construct the E. coli-Mycobacteria shuttle plasmid, the human interleukin-2 (IL-2) gene was electrophoreted into BCG with recombinant plasmid PZSIII-I and positive clones selected using polymerase chain reaction (PCR) technique. The expression of foreign gene of human IL-2 in BCG was identified by ELISA and SDS-PAGE. RESULT: Human IL-2 cytokine was steadily expressed in recombinant BCG and E. coli and could secrete outside the cell. CONCLUSION: M. bovis BCG recombinant constructed can produce and secrete the human IL-2. A secretion of the active cytokine was accomplished through the combined use of the BCG HSP65 promoter and a secretion signal from the BCG Ag-85B. The BCG HSP65 promoter is active in both BCG as well as E. coli which can not secrete foreign protein.
Using the specific monoclonal antibody of nm23H1 gene product and the immunohistochemical technique, we studied the expression of DNPK/nm23H1 and its correlation with lymph-node metastasis in nasopharyngeal carchnoma (NPC). The results revealed that, in a total of 31 cases of NPC, 13 showed positive expression with a positive rate of 41.9%. Among 21 cases of NPC with no lymph-node metastasis, 11 (52.3%) showed positive expression while only 20% (2/10) of those with lymph-node metastasis were positive, and there was significant difference between them (P < 0.01). Meantime the expression was very low or not detectable in the cancerous cells of the match lymph-node metastasis and all 10 cases showed negative expression. Statistical significant difference was found between the group of NPC with no metastasis and the group of lymph-node metastasis (P < 0.01). The above results indicated that high expression of DNPK/nm23H1 existed in the group of NPC with no metastasis while low expression existed in those with metastasis and lymph-node metastasis. It suggests that the expression of nm23H1 is inversely associated with lymph-node metastasis. The DNPK/nm23H1 may play a role in polymerization of microtubulin protein and participate in the process of cancer metastasis.
This paper presents a new kind of multivision glare tester. Its composition, function, hardware, software and application are introduced. The tester provides a practical tool for evaluating visual function change under glare-producing conditions.