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Biomedical subjects

X L Hu

Publications and source records attributed to X L Hu.

At least 37 records · Page 2Linked to original sources

[High performance thin layer chromatographic (HPTLC) determination of PAF in cerebrospinal fluid of patients with subarachnoid hemorrhage(SAH)].

The Platelet Activating Factor (PAF) is believed to be the major function in human after cerebral vascular spasm and cerebral ischemia. PAF has been found to participate in cerebral vascular spasm and cerebral ischemia by the basic and clinical study. The symptom of cerebral vascular spasm and cerebral ischemia has appeared with SAH. It has not been reported that the rule and change of PAF with SAH. In the present work, the concentration of PAF in human cerebral spinal fluid (CSF) with SAH were determined by high performance layer thin chromatography. The TLC plate was coated with high performance silica gel G using V(chloroform):V(methanol):V(water) = 65:35:6 as developing solvent. The PAF was determined by TLC scanning method and detected at 630 nm. The method was applied to determine the concentration of PAF in 16 CSF samples with SAH. The samples were collected in 1-3 d, 7-10 d and 14-21 d. CSF samples were deproteinized with methanol and chloroform. After centrifugation, the chloroform layer separated was dried at room temperature with nitrogen and stored under 20 degrees C in the refrigerator. The linear range of the method was 0.5-2.5 micrograms/L with regression coefficient of 0.9990. The lower limit of detection were 50 ng/L. The recovery of the method was 98.6%. The method enables a simple, rapid and reproducible quantification of PAF with SAH.

Chromatography, High Pressure Liquid↗

Structure of the terminal 300 kb of DNA from human chromosome 21q.

The most distal 300 kb of human chromosome 21q was cloned and mapped using telomeric yeast artificial chromosomes (YACs). The region contains low-copy subtelomeric repeats at the telomeric end, chromosome 21-specific sequences more centromerically, and the S100B gene at a distance of 100-140 kb from the chromosome terminus. RecA-assisted restriction endonuclease cleavage of genomic DNA showed that the cloned fragments correspond to telomere-terminal genomic DNA, and restriction enzyme mapping of the YACs shows that the smaller clone (175 kb) corresponds exactly to the telomeric end of the larger one (300 kb). PCR assays for 21q-specific markers were used to show that COL6A1, COL6A2, and LA161 were all outside of the subtelomeric region spanned by the YACs and thus at least 300 kb from the 21q terminus. The molecular probes provide telomeric closure for existing 21q maps.

Animals↗

Molecular cloning and RARE cleavage mapping of human 2p, 6q, 8q, 12q, and 18q telomeres.

Large terminal fragments of human chromosomes 2p, 6p, 8q, 12q, and 18q were cloned using yeast artificial chromosomes (YACs). RecA-assisted restriction endonuclease (RARE) cleavage analysis of genomic DNA samples from II unrelated individuals using YAC-derived probes confirmed the telomeric localizations of the half-YACs studied. The cloned fragments provide telomeric closure of maps for the respective chromosome arms and will supply the reagents needed for analyzing and sequencing these distal subtelomeric regions.

Base Sequence↗

[The relationship of apolipoprotein A-I and B-100 to cardiovascular diseases in aging males].

Apolipoprotein (apo) A-I and apo B-100 were examined in 378 aging males for studying the relationship of apolipoprotein to cardiovascular diseases. The results showed that apo A-I and apo B-100 were 148 +/- 30 mg/dl, 97.3 +/- 28 mg/dl in the healthy aged subjects. Patients with CHD had the higher level of apo B-100 (P < 0.01) than the controls. Apolipoprotein A-I was decreased (P < 0.05), but apo B-100 was increased (P < 0.01) in the patients with hypertension which indicated that the change of apolipoprotein could be a factor for patients with hypertension and sensitive to CHD. There were a positive correlation between apo B-100 and apo A-I cholesterol contents (r = 0.22, P < 0.05) as well as a negative correlation between apo A-I and fibrinogen (r = -0.2, P < 0.05) contents. Our data suggested that a higher content of apo A-I and lower content of apo B-100 might serve as the protective factors for CHD.

Aged↗

Physical analysis of the terminal 270 kb of DNA from human chromosome 1q.

DNA from three 1q44-derived human telomeric yeast artificial chromosome clones was analyzed using physical mapping methods. The smallest clone, yRM2004 (65 kb), corresponded exactly to the distal end of the largest clone, yRM2123 (270 kb). The third clone, yRM2192, overlapped with the proximal end of yRM2123 but not the distal end, suggesting that it is most likely a deletion artifact of a clone originally derived from a 1q telomere fragment. Data from fluorescence in situ hybridization analysis, restriction mapping, and RecA-assisted restriction enzyme cleavage experiments indicate that the molecular clone yRM2123 contains a 260-kb DNA fragment colinear with a genomic telomere-terminal fragment from 1q. yRM2123 contains low-copy subtelomeric and subterminal repeats at its distal end, single-copy DNA more centromerically, and a CG-rich region with homology to mouse DNA. Markers derived from this clone will allow telomeric closure of the physical and genetic linkage maps of human chromosome 1q.

Animals↗

Tumor necrosis factor-alpha mRNA and protein in endometrial tumors: analysis by in situ hybridization and immunocytochemistry.

Abnormal expression of polypeptide growth factors and their receptors is closely associated with tumorigenic transformation. In this study tumor necrosis factor-alpha (TNF-alpha) mRNA and protein were analyzed in polyps and proliferative lesions of endometrium as well as in low and high grade endometrial tumors by using in situ hybridization and immunocytochemistry. All samples contained products of the TNF-alpha gene. Histochemical scores (HS), which reflect the proportion of cells positive for TNF-alpha message or protein and the intensities of the signals, were higher for epithelial than for stromal cells. Benign lesions (endometrial polyps) contained little TNF-alpha mRNA or protein, whereas specific message was abundant in proliferative lesions (hyperplasia, adenofibroma). Although neoplastic cells in both low and high grade endometrial tumors contained TNF-alpha mRNA, two major differences were observed: HS for TNF-alpha mRNA were significantly less in low grade than in high grade neoplasms, and TNF-alpha message was restricted to the nucleus in low grade adenocarcinoma cells but was abundant in the cytoplasm of high grade tumor cells. In contrast to cells in benign and proliferative lesions, TNF-alpha protein scores in endometrial tumor cells were inversely rather than positively correlated with TNF-alpha mRNA scores. Collectively, the findings in this study are consistent with the postulate that TNF-alpha is useful to endometrial tumor cells and suggest that production may increase as cells diverge from normal.

Adenocarcinoma↗

Sequence organization of the human chromosome 2q telomere.

The terminal 240 kb of a human 2q telomere region was cloned in two overlapping yeast artificial-chromosomes (YACs). This DNA contains a region of low-copy subtelomeric repeats (within 50 kb of the 2q telomere), a segment of DNA duplicated on distal 8p23 (100 kb from the 2q telomere), and a region of single-copy DNA (230 kb from the 2q telomere). Two CpG islands are present in the DNA segment duplicated on distal 8p23. RecA-assisted restriction endonuclease cleavage of genomic DNA samples revealed a potential 55 kb chromosome length polymorphism at the 2q telomere. This work provides telomeric closure of maps for human chromosome 2q, demonstrates a novel, subtelomere-specific DNA duplication, and will permit detailed molecular and cytological studies of this human telomere region.

Base Sequence↗

[Clinical application of ultra-thin bronchofiberscope examination].

Ultra-thin bronchofiberscope examination has been performed in 73 persons including normal persons and patients with various bronchopulmonary disease. The main lesions of peripheral airways revealed by the examination were reddening, pigmentation, secretion, engorgement of blood vessels, mucosal pallor and edema. The result points out that this is a valuable new technique in the diagnosis of small airway lesions, especially the diffused inflammatory of small airways. In case of a localized lesion, because of the difficulty to precisely determine the site, the effectiveness of this examination is limited. We also briefly discussed the matters that one should pay attention to while performing this technique.

Adult↗

Combined therapy of captopril and spironolactone for refractory congestive heart failure.

It is traditionally considered that angiotensin--converting enzyme inhibitor (ACEI) and spironolactone could not be used simultaneously because of the assumed risk of hyperkalemia. However, despite ACEI therapy edema and congestive status remain in some of the patients with severe congestive heart failure (CHF). In order to seek an effective therapy for these patients, we observed the efficacy and safety of captopil plus spironolactone in patients with refractory CHF, with strict monitoring of renal function, serum and urine electrolytes and blood pressure (BP). Thirty-five patients with refractory CHF and New York Heart Association functional class IV without renal dysfunction, hypotension and hyperkalemia, whose plasma aldosterone (ALD) level in 88.6% of them was above normal value, were randomly assigned to group A (n = 16, captopril alone) and B (n = 19, captopril plus spironolactone) for a 4-week treatment. The dosage of both drugs was individually adjusted in time according to the results of serum potassium and renal function. The improvement in dyspnea--fatigue ratings, urinary volume and Na+/K+ ratio in group B was more significant than that in group A, and the plasma ALD level in group B decreased obviously while it remained high in group A after therapy. Two patients in group B who had had normal plasma ALD level with urinary Na+/K+ ratio > 1.0 before the therapy did not exhibit any clinical improvement. A strong negative correlation was found between plasma ALD level and urinary Na+/K+ ratio (correlation coefficient -0.689, P < 0.01). None of the patients had obvious hyperkalemia and hypoaldosteronism.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Normal distribution of tumor necrosis factor-alpha messenger ribonucleic acid and protein in the uteri, placentas, and embryos of osteopetrotic (op/op) mice lacking colony-stimulating factor-1.

In order to map mouse tumor necrosis factor-alpha (TNF) gene expression in detail and to determine whether transcription or translation of the TNF gene is regulated by uterine colony stimulating factor-1 (CSF-1), preimplantation embryos, oviducts, uteri, and uteroplacental units were studied in various strains of mice. These included homozygous osteopetrotic (op/op) female mice, which completely lack CSF-1, and heterozygous (+/op) females, which have normal levels of CSF-1. TNF mRNA was identified in all samples except preimplantation embryos by use of Northern blot hybridization or reverse transcriptase polymerase chain reaction. In situ hybridization and immunocytochemical experiments showed that the TNF gene was expressed in mouse oviduct and uterine epithelial cells, decidual cells, macrophage-like cells, placental trophoblast, and embryos. Despite an absence of CSF-1, TNF gene expression in the uteri, placentas, and embryos of op/op mothers did not differ in any major respect from expression in +/op or other strains of mice. The results of this study therefore indicate that the TNF gene is transcribed and translated in an ordered sequence through mouse gestation, and that maternal CSF-1 is not essential to expression of this cytokine gene. Collectively, these findings are consistent with a major role for TNF in mouse reproduction and development and with a potential compensatory function for this potent polypeptide factor in CSF-1 deficiency.

Animals↗

Differential distribution of interleukin-1 alpha and interleukin-1 beta proteins in human placentas.

Interleukin-1 alpha (IL-1 alpha) and interleukin-1 beta (IL-1 beta) were identified in samples of first trimester and term placentas by immunocytochemistry using two sets of polyclonal antibodies to recombinant IL-1. All samples contained the two species of IL-1, which predominated in different types of cells: IL-1 alpha was contained primarily in placental villous trophoblastic and stromal cells whereas IL-1 beta was localized to cells in fetal and maternal blood. With respect to the cellular localization of the two proteins, early and late gestation tissues were similar. Although these results were generally consistent for the two sets of antibodies, differences were noted in the binding patterns of the individual reagents. The findings in this study provide strong support for the postulate that IL-1 modulates placental development and suggest that IL-1 alpha may be the preferred species in extraembryonic tissues.

Blood Cells↗

Tumor necrosis factor-alpha gene expression in the tissues of normal mice.

Although TNF-alpha is traditionally associated with macrophage activation during neoplasia and acute inflammation, recent Northern blot hybridization studies indicate that gene expression occurs in the absence of pathology. In order to identify the cellular sources of endogenous message and protein, normal mouse tissues were tested for TNF-alpha mRNA using in situ hybridization and for the corresponding protein by immunocytochemistry. Unexpectedly, specific TNF-alpha message was readily detected in hepatocytes, kidney tubule epithelial cells, various populations of spleen cells and neurons. TNF-alpha protein was present in the same liver and kidney cells as those that contained TNF-alpha mRNA, was low in spleen cells, and was absent in neurons. These results suggest that cells other than macrophages are the major sources of TNF-alpha gene products in normal tissues, indicate that regulation is accomplished by more than one mechanism, and are consistent with the postulate that products of this gene contribute to normal physiological processes.

Animals↗

Tumor necrosis factor-alpha messenger ribonucleic acid and protein in human endometrium.

Although previous studies have shown that the tumor necrosis factor-alpha (TNF) gene is expressed in pregnancy decidua, it has not been determined whether this gene is transcribed or translated in the endometrium prior to implantation. To address this question and to identify cells expressing the TNF gene, samples of normal cycling human endometria were tested for TNF mRNA by in situ hybridization using a biotinylated antisense RNA probe, and the corresponding protein was localized in the same tissues by immunocytochemistry with a monoclonal antibody to TNF. Both TNF message and protein were identified in the endometrium throughout the menstrual cycle, and the same types of cells that contained transcripts also contained protein. As judged by the intensities of the hybridization signals, TNF mRNA increased during the proliferative phases, declined in the early secretory phase, and again rose during mid-to-late secretory phases, suggesting positive associations with levels of female sex hormones that show similar cyclic fluctuations. At the initiation of the cycle, transcripts were primarily localized to glandular epithelial cells whereas by the midproliferative phase, message was also present in stromal cells. Hybridization signals were consistently more intense in functionalis-region than in basalis-region stromal cells, and were frequently stronger in basalis-region glandular epithelia than in functionalis-region glands. These observations document that the TNF gene is expressed in normal cycling endometria, suggest that ovarian hormones may regulate TNF gene transcription, and identify differences in specific endometrial compartments. The findings are also consistent with the postulate that TNF is a local mediator of cellular communications in the human endometrium.

Adult↗

Functional interactions between the fusion protein and hemagglutinin-neuraminidase of human parainfluenza viruses.

The fusion glycoprotein (F) and hemagglutinin-neuraminidase (HN) genes of human parainfluenza virus type 2 (PI2) were molecularly cloned and expressed in HeLa-T4 cells by using the vaccinia virus-T7 transient expression system. Expression of the F and HN proteins was detected by using immunoprecipitation and surface immunofluorescence staining. Although the F protein was found to be cleaved into F1 and F2 and expressed on cell surfaces, no cell fusion was observed. However, cotransfection of the F-protein gene together with the P12 HN gene resulted in significant levels of cell fusion. Cell fusion was also observed when separate cell cultures were transfected with the HN and F genes and the F-expressing cells were mixed with the HN-expressing cells. Surprisingly, when the PI2 F protein was expressed together with the parainfluenza virus type 3 (PI3) HN protein, no fusion was detectable in the transfected cells. Similarly, no fusion was found upon coexpression of the PI2 HN and PI3 F proteins. However, coexpression of the PI3 F and HN proteins resulted in extensive cell fusion, which resembled the PI2 coexpression result. These results indicate that under the conditions used, the F protein is unable to cause fusion by itself and the HN protein provides a specific function in cell fusion which cannot be provided by another paramyxovirus attachment protein. Further, the results suggest that a type-specific functional interaction between the F and HN proteins is involved in mediating cell fusion.

Animals↗

Tumor necrosis factor alpha mRNA and protein are present in human placental and uterine cells at early and late stages of gestation.

Tumor necrosis factor alpha (TNF-alpha), a polypeptide that regulates cellular growth and modulates the synthesis of various cell surface and secreted molecules, has been identified in the pregnant uterus. To determine which specific cells transcribed and translated this gene, extraembryonic fetal tissues (placenta and membranes) and uterine tissue from early and late stages of gestation were analyzed for TNF-alpha mRNA by in situ hybridization using biotinylated antisense and sense TNF-alpha probes, and for immunoreactive TNF-alpha using two monoclonal antibodies. Tumor necrosis factor-alpha transcripts and protein were identified in both extraembryonic and maternal cells. In first-trimester placental villi, TNF-alpha mRNA was present in syncytiotrophoblast but was low to absent in cytotrophoblast and villous stromal cells. Decidual and epithelial cells in maternal tissues contained TNF-alpha transcripts. In term placentas, both syncytiotrophoblast and villous stromal cells contained TNF-alpha mRNA, and transcripts were present in maternal cells in the decidua adjacent to the extraplacental membranes. In both first-trimester and term tissues, coincident expression of TNF-alpha mRNA and immunoreactive TNF-alpha was demonstrated. The results of this study show that TNF-alpha is synthesized by cells in both extraembryonic membranes and maternal tissues during human gestation and that transcription in specific types of cells is influenced by gestational age. These observations are consistent with a major role for TNF-alpha in the dynamic developmental events of human pregnancy.

Delivery, Obstetric↗

Molecular cloning and sequence analysis of the fusion glycoprotein gene of human parainfluenza virus type 2.

A cDNA clone containing a 2.0-kb insert was identified as the human parainfluenza virus type 2 (PI2) fusion glycoprotein gene by hybridizing with a viral RNA probe and a synthetic oligonucleotide derived from a conserved sequence found in other paramyxovirus fusion protein genes. The complete nucleotide sequence of the glycoprotein gene was determined by the dideoxynucleotide sequencing procedure and found to contain a single, large open reading frame encoding a protein of 551 amino acids with a calculated molecular weight of 59,664. Comparison of the P12 fusion protein with those of other paramyxoviruses indicated similarities in overall length, N-terminal signal peptide sequence (amino acids 7 to 25), C-terminal membrane-spanning region (amino acids 486 to 513), and a highly conserved fusion sequence region at the N-terminus of the F1 subunit (amino acids 107 to 132).

Amino Acid Sequence↗