Hemoglobin H disease classification by isoelectric focusing: molecular verification of 110 cases from Thailand.
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Biomedical subjects
Publications and source records attributed to Wen Wang.
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Antimicrobial peptides secreted by the skin of many amphibians play an important role in innate immunity. From two skin cDNA libraries of two individuals of the Chinese red belly toad (Bombina maxima), we identified 56 different antimicrobial peptide cDNA sequences, each of which encodes a precursor peptide that can give rise to two kinds of antimicrobial peptides, maximin and maximin H. Among these cDNA, we found that the mean number of nucleotide substitution per non-synonymous site in both the maximin and maximin H domains significantly exceed the mean number of nucleotide substitution per synonymous site, whereas the same pattern was not observed in other structural regions, such as the signal and propiece peptide regions, suggesting that these antimicrobial peptide genes have been experiencing rapid diversification driven by Darwinian selection. We cloned and sequenced seven genes amplified from skin or liver genomic DNA. These genes have three exons and share the same gene structure, in which both maximin and maximin H are encoded by the third exon. This suggests that alternative splicing and somatic recombination are less likely to play a role in creating the diversity of maximins and maximin Hs. The gene trees based on different domain regions revealed that domain shuffling or gene conversion among these genes might have happened frequently.
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Embryonic stem cells (ES cells) can differentiate into endothelial cells and smooth muscle cells (SMCs), which participate in vascular angiogenesis. In this study, we differentiated mouse ES cells into Sca-1(+) cells, which have the potential to serve as vascular progenitor cells, and mapped their proteome by 2-DE using a pH 3-10 non-linear gradient and 12% SDS-polyacrylamide gels. A subset of 300 protein spots was analysed and mapped, with 241 protein spots being identified by their PMF using MALDI-TOF MS or by partial amino acid sequencing using MS/MS. Our protein map is the first of Sca-1(+) progenitor cells and will facilitate the identification of proteins differentially expressed during stem cell differentiation. The proteome of adult arterial SMCs is described in an accompanying paper (in this issue, DOI 10.1002/pmic.200402045). All data are made accessible on our website http://www.vascular-proteomics.com.
BACKGROUND: To investigate whether extrinsic antioxidant seleno-glutathione peroxidase mimic ebselen (PZ51) can protect endothelium and vascular structure of stroke-prone spontaneously hypertensive rats (SHRsp) during the chronic process of hypertension. METHODS: Twenty-two 8-week-old SHRsp were randomized into a PZ51 group and a control group, and administered by gavage for 6 weeks. We examined the level of nitric oxide (NO) and malonaldehyde (MDA) in plasma. The intima-media thickness (IMT) of the common carotid artery (CCA) was measured by an image-analysis system. The endothelium of the CCA was observed by scanning electron microscopy. The eNOS protein of the major artery was assayed by immunohistochemistry and western blotting. RESULTS: Compared with the control group, PZ51 decreased plasma MDA (7.88+/-1.06 vs 10.88+/-1.73 nmol/l, p<0.001) and increased plasma NO (40.02+/-9.74 vs 22.22+/-10.05 micromol/l, p<0.001), increased eNOS protein expression (8.25+/-2.36 vs 4.46+/-3.14, p=0.026), decreased IMT (69.85+/-5.47 vs 76.60+/-6.53 microm, p<0.05) significantly and alleviated the damage to the endothelium of the CCA. CONCLUSION: Administration of PZ51 for 6 weeks can protect the endothelium and inhibit vascular remodeling, maybe due to its suppression of lipid peroxide formation and increase in eNOS protein expression.
OBJECTIVE: Papillary loops in dermal papillae are believed to facilitate oxygen delivery to the germinal layer in the epidermis. The study aims to simulate the distribution of the oxygen partial pressure, pO2, in superficial layers of skin and to quantify oxygen supply from different sources. METHODS: A theoretical model that considered heterogeneity in tissue properties was developed based on the anatomical arrangement of the upper skin. Distribution of pO2 in tissues and in blood was solved numerically. Effects of the blood flow rate and the skin surface condition were investigated. RESULTS: Under normal blood flow rate and skin surface conditions, approximately one-quarter of the oxygen consumed in the upper skin came from papillary loops. Subpapillary plexus was the main supplier, which accounted for more than 60% of the total oxygen supply. Oxygen diffusion into the skin from the air was restricted to superficial regions of the germinal layer and accounted for approximately 10% of the total oxygen uptake. CONCLUSIONS: Papillary loops in dermal papillae facilitate oxygen delivery to the germinal layer from the circulation. The flow rate in papillary loops affects pO2 distribution in dermal papillae. In the epidermis, however, pO2 is strongly affected by skin surface conditions.
An increase in reactive oxygen species has been shown to play a role in perpetuating hypertension and cerebral injury in stroke-prone spontaneously hypertensive rats (SHRsp). Lipid peroxidation in the cerebral cortex is much more intense in SHRsp after establishment of severe hypertension as compared to that in normotensive Wistar-Kyoto rats (WKY). Cortical neurons from SHRsp are more vulnerable to hypoxia and hyponutritional conditions. We sought to investigate whether long-term administration of seleno-glutathione peroxidase mimic ebselen (PZ51) would have a protective effect on cortical neurons in SHRsp, and, if so, the possible mechanisms of this effect. Twenty-two 8-week-old SHRsp were randomized into a PZ51 group and control group. Age-matched WKY were used as normal controls. We examined the levels of malonaldehyde (MDA) and nitric oxide (NO) in the cerebral cortex (CC) homogenate, detected the three isoforms of nitric oxide synthase (NOS) by Western blotting, and examined cortical neurons by transmission electron microscopy. The results showed that PZ51 treatment significantly decreased both MDA and NO in the CC, inhibited inducible nitric oxide synthase (iNOS) protein expression, and alleviated the damage to cortical neurons compared to the findings for the control group. In conclusion, the present study showed that PZ51 administration suppressed lipid peroxidation and inhibited iNOS protein expression in CC homogenate, and it was suggested that these mechanisms may play a role in the protective effects of PZ51 on cortical neurons of SHRsp.
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OBJECTIVE: To research the effects of Polygala tenuifolia decoction on myoelectric activity of uterine smooth muscle and contractile activity of uterine smooth muscle strips of virginal rats. METHOD: To record the effects of P. tenuifolia decoction on myoelectric activity of uterine smooth muscle and contractile activity of uterine smooth muscle strips of virginal rats with biolap 410 biological system. Five blocking agents were used to study their mechanisms respectively. RESULT: Different dosages of water extract of P. tenuifolia (0.02, 0.04, 0.08 g x kg(-1)), could significantly potentiate uterine myoelectric activity and contractile activity of virginal rats. CONCLUSION: The effect of P. tenuifolia on myoelectric activity of uterine smooth muscle and contractile activity of uterine smooth muscle strips in rats may be mainly associated with H1 receptor, L-voltage-dependant calcium channels or prostaglandin synthese, its nothing to M receptor.
OBJECTIVE: Extraovarian peritoneal serous papillary carcinoma (EPSPC) is both histologically and clinically similar to stage III-IV ovarian papillary serous carcinoma (OPSC). The purpose of this study is to investigate the clinical findings, treatment, and outcome of EPSPC patients compared with stage III-IV OPSC patients. METHODS: The data of 12 EPSPC patients and 45 stage III-IV OPSC patients were retrospectively reviewed, comparing the characteristics on clinical presentation and treatment, sensitivity to first-line chemotherapy agents and survival. RESULTS: By analysis of patients' characteristics, presenting signs and symptoms, type and extent of surgery, tumor response to first-line chemotherapy, recurrence-free interval, recurrence site and serum CA-125 levels, no significant difference was observed between the EPSPC patients and stage III-IV OPSC controls. The prevailing presenting symptoms were abdominal mass and ascites. The mainstay of treatment was debulking surgery followed by adjuvant platinum-based chemotherapy. The complete clinical response of stage III-IV OPSC was 91.8% compared with 25.0% for women with EPSPC (P < 0.01). CONCLUSION: The clinical and surgical characteristics of EPSPC are similar to those of stage III-IV OPSC. When the same treatment strategy is applied, similar response and survival are expected in either condition.
OBJECTIVES: To investigate the effects of blood pressure lowering treatment on stroke recurrence among Chinese patients with previous cerebrovascular diseases. METHODS: Patients were eligible if they had a history of stroke or transient ischaemic attack (TIA) within the previous 5 years. Participants had no definite indication or contraindication for study drugs. There were no blood pressure entry criteria. This study was a large-scale randomised, double-blind, placebo controlled clinical trial. After 4 weeks run-in period, 1520 randomised patients received either ACE inhibitor-perindopril (+ diuretic-indapamide) treatment or matching placebo for 4 years. The primary study outcome was stroke event. Secondary outcomes included cardiovascular death, myocardial infarction, all-cause death and blood pressure. RESULTS: Ssven hundred and sixty-two were assigned active treatment and 758 assigned placebo. The characteristics of randomised patients of active and control groups were similar. In active and control groups: 70.8% and 70.5% were male, 93.8% and 93.4% had a history of stroke (haemorrhagic or cerebral infarction). Baseline mean blood pressure in the active and control groups were 145.3 +/- 20.2/86.8 +/- 11.1 mm Hg and 145.3 +/- 20.3/87.2 +/- 10.8 mm Hg, and mean age were 63.9 +/- 7.5 and 63.8 +/- 7.7 years, respectively. Blood pressure in those assigned active treatment was reduced on average 14/6 mm Hg more than placebo at 4 years. During double-blind treatment, active treatment reduced stroke recurrence (8.8% vs 19.4%) by 55% (P < 0.001), myocardial infarction (1.4% vs 2.8%) by 48% (P = 0.070), cardiovascular mortality (3.6% vs 6.6.%) by 45% (P = 0.010), and all-cause mortality (6.3% vs 9.8%) by 36% (P = 0.010). These benefits were achieved similarly in all subgroups: male or female, middle-aged or elderly, with or without hypertensive, cerebral infarct or haemorrhagic stroke. CONCLUSION: 4 years of blood pressure lowering treatment was beneficial in Chinese patients with previous cerebrovascular diseases.
OBJECTIVE: We developed a solid phase adsorption gas sampler, which is suitable for collection and analysis of indoor air formaldehyde. METHODS: This solid phase adsorption gas sampler is composed of two parts, a support part made of a glass tube covered silica-gel stopper on ports, an absorbent part in glass tube made of surface denatured synthetic fibre coated formaldehyde absorbent. We evolved its performance, and established an method for sampling and determination formaldehyde in indoor air with the gas sampler. RESULTS: For the solid phase adsorption gas sampler, the collection efficiency was 98.7%, and the desorption efficiency was 98.6%. The variation coefficients of sampling was 4.02% (n = 10). The above method was compared with the standard method (GB/T 16129-1995 ), the results showed no significant difference between the two methods. CONCLUSION: This solid phase adsorption gas sampler can be used for sampling and determination formaldehyde in indoor air.
OBJECTIVE: To study the feasibility of using free fetal DNA in maternal plasma for non-invasive prenatal diagnosis. METHODS: Maternal DNA extracted from plasma samples of 10 pregnant women at early pregnancy, medium pregnancy and late pregnancy and their husband's DNA isolated from whole blood samples were used to detect genotype by multiplex fluorescent PCR at nine different polymorphic short tandem repeat(STR) loci. RESULTS: Fetus-specific alleles were found in maternal plasma samples studied. By the application of these polymorphic short repeat sequences, every pregnant women/husband pair was informative in at least three of nine loci. Paternally inherited fetal alleles were detected in 23 of 30 plasma samples. They are 6/10 cases in early pregnancy, 8/10 cases in middle pregnancy and 9/10 cases in late pregnancy respectively. CONCLUSION: Fluorescent multiplex PCR can be used for amplification of male and female fetal STRs in maternal plasma to obtain genetic information, which may have implication for non-invasive prenatal diagnosis of certain hereditary diseases independent of the fetal sex.
We report a draft sequence for the genome of the domesticated silkworm (Bombyx mori), covering 90.9% of all known silkworm genes. Our estimated gene count is 18,510, which exceeds the 13,379 genes reported for Drosophila melanogaster. Comparative analyses to fruitfly, mosquito, spider, and butterfly reveal both similarities and differences in gene content.
The reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to examine the changes of the expression of 5-hydroxytryptamine (5-HT) receptors in the rat lumbar dorsal root ganglion (DRG) following subcutaneous bee venom (BV) injection into the plantar surface of the unilateral hindpaw. In the DRG ipsilateral to the BV injection, significant increase of mRNA levels of 5-HT(1A), 5-HT(1B), 5-HT(2A) and 5-HT(3) receptor subtypes were observed at 1 and 4h after the BV injection, while increase of 5-HT(2C), 5-HT(4), 5-HT(6) and 5-HT(7) receptor subtype mRNAs was detected at 4h only. No such changes were observed in the expressions of 5-HT(1D), 5-HT(1F) and 5-HT(5A) receptor subtype mRNAs. Upregulation of 5-HT(1A), 5-HT(1B) and 5-HT(2A) receptor subtype mRNAs was also observed in the contralateral DRG at 4 h. The presence of 5-HT(1E), 5-HT(2B) and 5-HT(5B) receptor subtype mRNAs was not detected in the rat DRG. The present results suggest that different sets of 5-HT receptor subtypes work at different stages of the inflammatory pain induced by subcutaneous BV injection.
In the present study, the effects of 17beta-estradiol (E2) and phytoestrogen alpha-zearalanol (ZAL) on tissue factor (TF) in plasma of ovariectomized (OVX) rats and human umbilical vein endothelial cells (HUVECs) were investigated. OVX rats were injected intramuscularly twice a week with E2 (1mg/kg) or ZAL (1mg/kg) for five weeks. The concentrations of plasma E2 were measured by radioimmunoassay. The TF levels of plasma were assayed by enzyme-linked immunosorbent assay (ELISA). HUVECs were exposed to E2 or ZAL for 48 h. TF protein and mRNA were assayed by ELISA and RT-PCR, respectively. Nuclear proteins of HUVECs opposed to E2 or ZAL for 8 h were extracted to assay transcription factors activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) by Western blot. Both E2 and ZAL were found to decrease plasma TF levels of OVX rats significantly. E2 or ZAL were also found down-regulate the expression of TF protein and mRNA in HUVECs. Moreover, both E2 and ZAL might decrease AP-1 and NF-kappaB levels. These results suggest that both E2 and ZAL have inhibitory effects on TF in plasma of OVX rats and HUVECs, which may then contribute to their protective effects on cardiovascular diseases.
Deoxynegamycin (1b) is a protein synthesis inhibitor with activity against Gram-negative (GN) bacteria. A series of conformationally restricted analogs were synthesized to probe its bioactive conformation. Indeed, some of the constrained analogs were found to be equal or better than deoxynegamycin in protein synthesis assay (1b, IC(50)=8.2 microM; 44, IC(50)=6.6 microM; 35e(2), IC(50)=1 microM). However, deoxynegamycin had the best in vitro whole cell antibacterial activity (Escherichia coli, MIC=4-16 microg/mL; Klebsiella pneumoniae, MIC=8 microg/mL) suggesting that other factors such as permeation may also be contributing to the overall whole cell activity. A new finding is that deoxynegamycin is efficacious in an E. coli murine septicemia model (ED(50)=4.8 mg/kg), providing further evidence of the favorable in vivo properties of this class of molecules.
Members of the DExH/D superfamily of nucleic acid-activated nucleotide triphosphatases are essential for virtually all aspects of RNA metabolism, including pre-messenger RNA splicing, RNA interference, translation, and nucleocytoplasmic trafficking. Physiological substrates for these enzymes are thought to be regions of double-stranded RNA, because several DExH/D proteins catalyze strand separation in vitro. These "RNA helicases" can also disrupt RNA-protein interactions, but it is unclear whether this activity is coupled to duplex unwinding. Here we demonstrate that two unrelated DExH/D proteins catalyze protein displacement independently of duplex unwinding. Therefore, the essential functions of DExH/D proteins are not confined to RNA duplexes but can be exerted on a wide range of ribonucleoprotein substrates.