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Biomedical subjects

A Hu

Publications and source records attributed to A Hu.

At least 19 recordsLinked to original sources

Characterization of phi8, a bacteriophage containing three double-stranded RNA genomic segments and distantly related to Phi6.

The three double-stranded RNA genomic segments of bacteriophage Phi8 were copied as cDNA, and their nucleotide sequences were determined. Although the organization of the genome is similar to that of Phi6, there is no similarity in either the nucleotide sequences or the amino acid sequences, with the exception of the motifs characteristic of viral RNA polymerases that are found in the presumptive polymerase sequence. Several features of the viral proteins differ markedly from those of Phi6. Although both phages are covered by a lipid-containing membrane, the protein compositions are very different. The most striking difference is that protein P8, which constitutes a shell around the procapsid in Phi6, is part of the membrane in Phi8. The host attachment protein consists of two peptides rather than one and the phage attaches directly to the lipopolysaccharide of the host rather than to a type IV pilus. The host range of Phi8 includes rough strains of Salmonella typhimurium and of pseudomonads

Amino Acid Motifs↗

The biosynthetic gene cluster for the microtubule-stabilizing agents epothilones A and B from Sorangium cellulosum So ce90.

BACKGROUND: Epothilones are produced by the myxobacterium Sorangium cellulosum So ce90, and, like paclitaxel (Taxol((R))), they inhibit microtubule depolymerisation and arrest the cell cycle at the G2-M phase. They are effective against P-glycoprotein-expressing multiple-drug-resistant tumor cell lines and are more water soluble than paclitaxel. The total synthesis of epothilones has been achieved, but has not provided an economically viable alternative to fermentation. We set out to clone, sequence and analyze the gene cluster responsible for the biosynthesis of the epothilones in S. cellulosum So ce90. RESULTS: A cluster of 22 open reading frames spanning 68,750 base pairs of the S. cellulosum So ce90 genome has been sequenced and found to encode nine modules of a polyketide synthase (PKS), one module of a nonribosomal peptide synthetase (NRPS), a cytochrome P450, and two putative antibiotic transport proteins. Disruptions in the genes encoding the PKS abolished epothilone production. The first PKS module and the NRPS module are proposed to co-operate in forming the thiazole heterocycle of epothilone from an acetate and a cysteine by condensation, cyclodehydration and subsequent dehydrogenation. The remaining eight PKS modules are responsible for the elaboration of the rest of the epothilone carbon skeleton. CONCLUSIONS: The overall architecture of the gene cluster responsible for epothilone biosynthesis has been determined. The availability of the cluster should facilitate the generation of designer epothilones by combinatorial biosynthesis approaches, and the heterologous expression of epothilones in surrogate microbial hosts.

Anti-Bacterial Agents↗

Simultaneous determination of trace benzodiazepines from drinks by using direct electrospray probe/mass spectrometry (DEP/MS).

This study presents a novel means of rapidly determining benzodiazepines in various drinks. Electrospray mass spectrometry with a direct probe design is used as the analytical instrument. Samples are treated only by a simple liquid-liquid extraction prior to direct electrospray probe/mass spectrometry (DEP/MS) analysis. The proposed method provides a relatively easy and efficient means of identifying the drugs in a very short time interval. On average, sample analysis is completed in less than five minutes including sample pretreatment. It is especially useful in forensic science since the technology can provide a rapid identification for unknown samples. Thus, the method proposed herein is highly appropriate for rapidly screening a large number of samples.

Beer↗

[Genetic linkage of polymorphisms of type I angiotensin II receptor gene to Chinese Han hypertension]

OBJECTIVE: To observe the CA repeat polymorphism of the type I angiotensin II receptor gene on Chinese Han people, and evaluate the genetic linking to hypertension. METHODS: The CA repeat polymorphism at 3'-flanking region of AT1R gene was studied by amplified fragment length polymorphism (Amp-FLP ) in Chinese Han people from Shandong and Xi'an. RESULTS: There were 9 alleles ranging from 130bp to 146bp in all subjects. The allele frequencies were 0.01 to 0.38, and allele A4 was more frequent in the control. The heterozygosity of CA repeat locus was 0.789, the polymorphic information content, 0.746. There was no significant difference in distribution of CA repeat polymorphism genotype between different regions in same ethnic (P>0.05), but the allele frequency of CA repeat polymorphism in hypertensive group was significantly different from that of control (P<0.05). There was no detectable distortion from Hardy-Weinberg equilibrium in both groups. CONCLUSION: The results suggest that CA repeat polymorphism of AT1R gene be associated with Chinese Han hypertensive people.

Journal Article↗

[Genetic linkage of polymorphisms of type I angiotensin II receptor gene to Chinese Han hypertension].

OBJECTIVE: To observe the CA repeat polymorphism of the type I angiotensin II receptor gene on Chinese Han people, and evaluate the genetic linking to hypertension. METHODS: The CA repeat polymorphism at 3'-flanking region of AT1R gene was studied by amplified fragment length polymorphism (Amp-FLP ) in Chinese Han people from Shandong and Xi'an. RESULTS: There were 9 alleles ranging from 130bp to 146bp in all subjects. The allele frequencies were 0.01 to 0.38, and allele A4 was more frequent in the control. The heterozygosity of CA repeat locus was 0.789, the polymorphic information content, 0.746. There was no significant difference in distribution of CA repeat polymorphism genotype between different regions in same ethnic (P>0. 05), but the allele frequency of CA repeat polymorphism in hypertensive group was significantly different from that of control (P<0.05). There was no detectable distortion from Hardy-Weinberg equilibrium in both groups. CONCLUSION: The results suggest that CA repeat polymorphism of AT1R gene be associated with Chinese Han hypertensive people.

Adult↗

Recurrent acyclovir-resistant herpes simplex in an immunocompromised patient: can strain differences compensate for loss of thymidine kinase in pathogenesis?

To investigate how acyclovir-resistant (ACVr) herpes simplex virus (HSV) evades drug therapy and causes disease, HSV-1 isolates from a bone marrow transplant (BMT) patient were studied. The patient developed ACVr disease after an initial BMT and, following a second BMT, reactivated ACVr HSV despite high-dose acyclovir prophylaxis. ACVr isolates from each episode contained the same point mutation in the viral thymidine kinase (tk) gene, documenting the emergence, latency, and reactivation of this mutant. The mutants were exceedingly impaired for TK activity in sensitive enzyme, plaque autoradiography, and drug-susceptibility assays. Nevertheless, these mutants and a tk deletion mutant constructed in the same genetic background reactivated from latency in mouse trigeminal ganglia, in contrast to similar mutants from laboratory strains. It is hypothesized that alleles in the clinical isolate compensate for the loss of TK in this animal model. Such genetic variability may be important for ACVr disease in humans.

Acyclovir↗

Cephalometric and occlusal changes following maxillary expansion and protraction.

A prospective clinical trial was conducted to determine the cephalometric and occlusal changes following maxillary expansion and protraction. Twenty Southern Chinese patients (eight males and 12 females with a mean age of 8.4 +/- 1.8 years) with skeletal Class III malocclusions were treated consecutively with maxillary expansion and a protraction facemask. Growth adaptation of these patients was followed for 2 years after removal of the appliances and compared with a control group of subjects with no treatment. Lateral cephalometric radiographs were used to quantify the skeletal and dental changes before treatment (T1), immediately after treatment (T2) and 2 years after removal of appliances (T3). With 8 months of treatment (T2-T1), overjet was overcorrected from a -2.0 to 3.5 mm. The maxilla moved forwards by an average of 2.1 mm and the molar relationship was improved to a Class I dental arch relationship. The palatal and occlusal planes were tilted upward 1.0 and 2.0 degrees, respectively. Two years following removal of the appliances (T3-T2), a positive overjet was maintained in 18 out of 20 patients. The maxilla continued to move forwards in the treated subjects similar to the controls. The mandible outgrew the maxilia. In most instances, dental compensation with proclination of the maxillary incisors was observed. The palatal plane returned to pre-treatment value. The occlusal plane continued to tilt upward due to eruption of the molars and proclination of the incisors. Analysis of dental casts showed a significant increase in maxillary intercanine (2.2 mm) and intermolar widths (2.3 mm) with 7 days of rapid palatal expansion followed by maxillary protraction. The percentage relapse in maxillary intermolar widths was 30-45 per cent after 1 year, in most cases with minimal retention. In the mandibular arch, the concurrent increase in intermolar width (2.3 mm) was primarily due to buccal uprighting of the posterior molars when the maxilla was protracted into a Class I skeletal relationship and was stable after 1 year. The results of this study indicate stability of orthopaedic treatment of Class III malocclusions directed at the maxilla. Despite some relapse, a net improvement in maxillomandibular relationship and a positive overjet was maintained in 18 out of 20 patients at the end of the follow-up period.

Cephalometry↗

Human cytomegalovirus mutant with sequence-dependent resistance to the phosphorothioate oligonucleotide fomivirsen (ISIS 2922).

A human cytomegalovirus mutant that was isolated for resistance (10-fold) to the antisense oligonucleotide fomivirsen (ISIS 2922) exhibited cross-resistance to a modified derivative of fomivirsen with an identical base sequence but little or no resistance to an oligonucleotide with an unrelated sequence. No changes in the mutant's DNA corresponding to the fomivirsen target sequence were found.

Antiviral Agents↗

[The mechanism of hemorrhage following phosphorus burns and the effect of calcium gluconate].

OBJECTIVE: To investigate the mechanism of hemorrhagic tendency after phosphorus poisoning. METHODS: 45% phosphoric acid and 20 mg/cm2 phosphorus were used to produce burn injury in rabbits. The total content of phosphorus remaining on the wound and 7-day mortality were similar in both groups. RESULTS: The results showed that plasma free calcium contents were significantly lower in both phosphorus groups than healthy and thermal burn controls (0.80 vs 1.40 mol/l, P < 0.01). The free calcium content of platelet was lowered to 116-140 nmol/l, which was significantly lower than that of healthy control (285 nmol/l) and thermal injury group (480-504 nmol/l). Concomitantly, the platelet aggregation rate was lowered to 1.29% and 10.78% in phosphoric acid group and phosphorus group, respectively, while it was 67.01% in healthy control and 82.1%-84.9% in thermal injury group; the difference was statistically significant. CONCLUSION: Calcium therapy brought back intracellular and extracellular free calcium contents to normal in phosphorus burn group. Also platelet aggregation rate approached normal level after calcium therapy. The results suggest that the bleeding tendency after phosphorus burn is due to combination of phosphorus with intracellular and extracellular calcium of platelets, thus inhibiting aggregation of platelets.

Animals↗

[Analysis of surface markers of long-term cultured human umbilical cord blood hemopoietic cells].

OBJECTIVE: To explore the optimal time for in vitro expansion and transplantation of umbilical cord blood hemopoietic cells. METHODS: Hemopoietic cells from human umbilical cord blood were cultured with SCF, rIL-3, rIL-6 and rIL-1 beta in a long term culture system and changes of cellular surface markers were observed. RESULTS: CD3+ and CD20+ lymphocytes increased at the 7th day, and gradually decreased to the lowest level after the 14th day. CD34 positive cells increased after the 7th day, and reached the highest level at the 14th day, then decreased gradually but maintained at a higher level than that at the 7th day. CD33 positive cells increased at the 7th day, and reached the highest level from the 14th to the 21st day, then decreased after the 28th day. CD42b positive cells began to increase at the 21st day, reached the peak at the 28th day, and still maintained higher level at the 35th day. HLA-DR positive cells increased from the 14th to the 21st day. CONCLUSION: The culture system containing the four growth factors is able to expand hemopoietic cells from human umbilical cord blood and keeps them intact. From the 14th to the 21st day, the number of hemopoietic cells reached the highest level, and is suitable for transplantation.

Antigens, CD20↗

[Deletion polymorphism in the angiotensin converting enzyme gene associated with essential hypertension in Hans Chinese population].

OBJECTIVE: To investigate whether the polymorphism of the angiotensin-converting enzyme(ACE) gene is associated with essential hypertension in Chinese. METHODS: A case-control study was carried out using 134 hypertensive (HT) and 165 normotensive (NT) subjects. The insertion/deletion (I/D) polymorphism of ACE gene was detected by polymerase chain reaction (PCR). RESULTS: The difference of genotype and derived allele frequencies for deletion of ACE gene between hypertensive and normotensive subjects were statistically significant(i.e. 51.9% vs 15.7 and 0.69 vs 0.31 for hypertensive and normotensive respectively, P < 0.05). CONCLUSIONS: The possession of D/D homozygote genotype of the ACE gene might be a marker for genetic susceptibility to Hans hypertension in Chinese.

Adult↗

Immunoregulatory changes in Kawasaki disease.

Kawasaki disease (KD) is an acute vasculitis of unknown etiology, occurring in young children and treated with intravenous gamma globulin (IVIG) to prevent significant cardiac morbidity and mortality. We studied KD patients pre- and post-IVIG therapy and at >40 days posttherapy, additionally comparing them with matched pediatric control patients and parents. Using three-color flow cytometry, we examined immune changes in KD, especially previously unassessed markers of T-lymphocyte activation, memory, and adhesion. The percentage of cells positive for CD19, CD25, CD38, and CD71 was significantly lower during convalescence compared with pre-IVIG (medians: CD19, 18% vs 26%, P = 0.0004; CD25, 6% vs 9% for CD3(+) cells, P = 0.0074; CD38, 78% vs 89% for CD8(+) cells, P = 0.0015; CD71, 1% vs 6% for CD4(+) cells, P = 0.0024). The proportion of CD3(+) cells increased (medians: CD3, 66% vs 45%, P < 0.0001). Values for all parameters varied greatly pre- and post-IVIG, but not in a consistent direction. The sole patient with cardiac abnormalities had the greatest pre-/post-IVIG variability. These changes support the involvement of T-lymphocytes in the acute KD vasculitic process. They also suggest that T-lymphocytes involved in endothelial damage during acute KD may be subsequently removed or eliminated from the peripheral blood.

Antigens, CD↗

Selective expression of a subset of measles virus receptor-competent CD46 isoforms in human brain.

The human cell surface protein CD46 is the main measles virus (MV) receptor. We analyzed the CD46 isoforms expressed in the brain of three patients who died with persistent MV infections and in an unaffected brain. Complete CD46 cDNAs were produced and found to code exclusively for CD46 isoforms with cytoplasmic tail 2. Selective expression of tail 2 isoforms was shown in a second control brain by Western blots with antibodies specific for each of the cytoplasmic tails. Binding of purified MV particles and virus-dependent cell fusion were tested after transient expression of brain-derived CD46 proteins in mouse cells. All the brain-derived proteins mediated MV binding and virus-dependent fusion. Isoforms containing both serine/threonine/proline (STP)-rich domains were more active in virus binding, whereas isoforms with only one STP domain were more efficient in mediating fusion.

Amino Acid Sequence↗

Construction of a 750-kb bacterial clone contig and restriction map in the region of human chromosome 21 containing the progressive myoclonus epilepsy gene.

The gene responsible for progressive myoclonus epilepsy of the Unverricht-Lundborg type (EPM1) is located on human chromosome 21q22.3 in a region defined by recombination breakpoints and linkage disequilibrium. As part of an effort to clone the EPM1 gene on the basis of its chromosomal location, we have constructed a 753-kb bacterial clone contig that encompasses the region containing the gene. Because DNA markers from the region did not identify intact yeast artificial chromosome (YAC) clones after screening several libraries, we built the contig from cosmid clones and used bacterial artificial chromosome (BAC) and bacteriophage P1 clones to fill gaps. In addition to constructing the clone contig, we determined the locations of the EcoRI, SacII, EagI, and NotI restriction sites in the clones, resulting in a high-resolution restriction map of the region. Most of the contig is represented by a level of redundancy that allows the orders of most restriction sites to be determined, provides multiple data points supporting the clone orders and orientations, and allows a set of clones with a minimum degree of overlap to be chosen for efficient additional analysis. The clone and restriction maps are in excellent agreement with maps generated of the region by other methods. These ordered bacterial clones and the mapping information obtained from them provide valuable reagents for isolating candidate genes for EPM1, as well as for determining the nucleotide sequence of a 750 kb region of the human genome.

Base Sequence↗

[Investigation of soluble tumor necrosis factor receptor in patients with active tuberculosis].

It is assumed that soluble tumor necrosis factor receptor I and II (sTNF-RI, II) play important roles in the regulation of tumor necrosis factor alpha (TNF-alpha) activity. We measured the levels of circulating sTNF-R in patients with active pulmonary tuberculosis (n = 31) and the correlation between TNF-alpha and sTNF-R in serum level was investigated. We also compared sTNF-R levels before and after the treatment in 7 cases. Significant increase of circulating sTNF-R were found in patients with tuberculosis compared with the normal controls (n = 28) (p < 0.01). Moreover, significant positive correlations were found between TNF-alpha and sTNF-R I and II (r = 0.520, r = 0.553) in serum comparing sTNF-R levels before and after the treatment for patients with tuberculosis, significant fall was found in sTNF-R I, but not in sTNF-R II. As a result, it is suggested that sTNF-R regulates TNF-alpha activity in patients with tuberculosis, and that sTNF-R I levels could be used as one of the indices to evaluate the clinical activity of tuberculosis.

Adult↗

Field evaluation of the QBC technique for rapid diagnosis of vivax malaria.

The QBC (quantitative buffy coat) technique was compared with that of the Giemsa-stained thick blood film (GTF) under field conditions in Junlian and Mingshan counties, Sichuan, China, for rapid diagnosis of vivax malaria. Blood samples were collected from 364 volunteer villagers, and each sample was examined with both the QBC and GTF techniques. For each GTF sample (10 microliters of blood), as many as 300 oil-immersion fields were examined; each QBC tube was inspected for up to 5 minutes. The GTF technique resulted in 86 positive blood samples and 278 negative; the QBC technique indicated 89 positive and 275 negative samples. Relative to the results obtained with GTF, the QBC technique had a sensitivity and specificity of 87.2% and 95.0%, respectively; concordance between the tests was 93.1%. The median time-to-positive diagnosis with the QBC technique (1.12 min) was 11% of that with GTF. The distribution of different developmental stages of Plasmodium vivax parasites was also examined in the centrifuged QBC tubes: all stages except schizonts could be found in the lower part of the platelet zone (the interphase between the monocyte and platelet layers), especially ring forms.

Azure Stains↗