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Biomedical subjects

S Zheng

Publications and source records attributed to S Zheng.

At least 73 records · Page 4Linked to original sources

[Mapping the interaction site of Rpb2 and Rpb3 subunit of fission yeast RNA polymerase II].

To map the interacting site of subunit Rpb2 to subunit Rpb3 of RNA polymerase II in fission yeast Schizosaccharomyces pombe, the yeast two-hybrid system was employed in this paper to screen the interacting clones between Rpb2 and Rpb3.4 fragments of Rpb2 cDNA were cloned into the Ga14 BD vector pAS2. The 4 clones were named as pAS2 Rpb2-1, 2-2, 2-3 and 2-4, respectively. The complete cDNA of Rpb3 was cloned into the Gal 4 AD vector pGADGH. The clone was named as pGADGH Rpb3. The two-hybrid plasmids pGADGH Rpb3 and pAS2Rpb2-1, 2-2, 2-3 or 2-4 respectively were cotransformed into host cell yeast Y190. The interaction positive cotransformants were identified by beta-gal activity assay. The beta-gal positive cotransformants were selected from pGADGH Rpb3 and pAS2Rpb2-4 two-hybrid system. DNA sequencing and alignment results showed that the interacting site of Rpb2 to Rpb3 located within the fragment from base 2701 to 2966 of Rpb2 cDNA, or within the C-termini polypeptide from amino acid 902 to 989 of Rpb2 protein.

Amino Acid Sequence↗

[Clinical study on the efficacy of tranexamic acid in reducing postpartum blood lose: a randomized, comparative, multicenter trial].

OBJECTIVE: To study the efficacy and safety of Transamin (tranexamic acid) in reducing postpartum blood loss. METHODS: Four hundred primipara with term singleton pregnancy, vertex presentation, spontaneous delivery were enrolled. Ten U Oxytocin was injected intravenously immediately after the delivery of fetal shoulders in the second stage of labor. Then all the puerperants were randomly assigned to 4 groups. Group I (n = 94): Transamin 1.0 g was injected i.v.; Group lI (n = 92): Transamin 0.5 g i.v.; Group III (n = 92): aminomethylbenzoic acid 0.5 g i.v. was given; Group IV (n = 87): no treatment. Vaginal bleeding was precisely collected, and examined immediately after the expulsion of placenta and from placenta expulsion till 2 hours after delivery. The amount of blood loss was measured by both methods of weight and volume. RESULTS: There were no significant differences of blood loss immediately after the expulsion of placenta among the 4 groups (P > 0.05). For the average blood loss at 2 hour postpartum, it was 129.7 ml, 133.9 ml, 168.5 ml and 178.2 ml for group I, II, II and IV respectively, while the total blood loss for the 4 groups was 243.3 ml, 242.9 ml, 308.1 ml, and 314.8 ml respectively. The average blood loss of group I and II was significantly less than group III and IV (P < 0.01), however, there was no significant difference between group I and group II (P > 0.05). The occurrences of postpartum hemorrhage (blood lose > or = 400 ml) were 6.4%, 13.3%, 20.7% and 25.3% for group I, II, III and IV respectively. There was no major adverse effects appeared. CONCLUSIONS: Transamin is efficient and safe in reducing the postpartum blood loss. 1.0 g of Transamin has the best efficacy, and 0.5 g of Transamin followed.

Antifibrinolytic Agents↗

Molecular mechanisms of increased nitric oxide (NO) in asthma: evidence for transcriptional and post-translational regulation of NO synthesis.

Evidence supporting increased nitric oxide (NO) in asthma is substantial, although the cellular and molecular mechanisms leading to increased NO are not known. Here, we provide a clear picture of the events regulating NO synthesis in the human asthmatic airway in vivo. We show that human airway epithelium has abundant expression of NO synthase II (NOSII) due to continuous transcriptional activation of the gene in vivo. Individuals with asthma have higher than normal NO concentrations and increased NOSII mRNA and protein due to transcriptional regulation through activation of Stat1. NOSII mRNA expression decreases in asthmatics receiving inhaled corticosteroid, treatment effective in reducing inflammation in asthmatic airways. In addition to transcriptional mechanisms, post-translational events contribute to increased NO synthesis. Specifically, high output production of NO is fueled by a previously unsuspected increase in the NOS substrate, l -arginine, in airway epithelial cells of asthmatic individuals. Finally, nitration of proteins in airway epithelium provide evidence of functional consequences of increased NO. In conclusion, these studies define multiple mechanisms that function coordinately to support high level NO synthesis in the asthmatic airway. These findings represent a crucial cornerstone for future therapeutic strategies aimed at regulating NO synthesis in asthma.

Adult↗

Wnt-1 inhibits nerve growth factor-induced differentiation of PC12 cells by preventing the induction of some but not all late-response genes.

The vertebrate Wnt-1 proto-oncogene is expressed transiently in embryonic brain and functions in the development of the central nervous system and neural crest. The role of Wnt-1 in neural crest development appears to be to increase the number of certain progenitor cells by preventing their premature differentiation. To study the mechanism by which this transient Wnt-1 expression inhibits differentiation we have constructed PC12 pheochromocytoma cells in which Wnt-1 expression levels were controlled by use of a tetracycline-responsive transactivator. Induction of Wnt-1 expression by tetracycline withdrawal was followed by activation of the Wnt-1 signalling pathway as shown by activation of the Lef-1/Tcf transcription factor. Wnt-1 expression by these cells resulted in reversible inhibition of NGF-induced neurite outgrowth, but it did not adversely affect the maintenance of previously formed NGF-induced neurites. Wnt-1 expression also partially blocked the ability of NGF to decrease the rate of cell multiplication. Wnt-1 decreased the NGF-induced expression of the late-response gene SCG10 but not of the immediate early genes, fos, Nur77 and UPAR (urokinase-type plasminogen activator receptor) nor of the late-response genes GAP-43 and collagenase. The Wnt-1 expressing PC12 cells multiplied at a greater rate when they expressed Wnt-1 than they did in the absence of Wnt-1 expression, a result that is consistent with the proposal that Wnt-1 may also act as a mitogen.

ATP-Binding Cassette Transporters↗

Histologic signatures of thermal injury: applications in transmyocardial laser revascularization and radiofrequency ablation.

BACKGROUND AND OBJECTIVE: Cardiac treatments such as transmyocardial laser revascularization and radiofrequency ablation cause thermal injury. We sought to provide quantitative histologic methods of assessing such injury by using the inherent birefringence of cardiac muscle and collagen; specifically, to exploit the connection between thermal injury and the loss of birefringence. STUDY DESIGN/MATERIALS AND METHODS: We quantified tissue birefringence changes in vitro for temperatures up to 130 degrees C. This information was used to assess thermal injury associated with myocardial channels made in vitro. We then measured in vivo cardiac injury 30 minutes and 3 days after radiofrequency exposure. RESULTS: Birefringence decreased above 60 degrees C for muscle and above 70 degrees C for collagen. Temperatures above 80 degrees C were associated with collagen fiber straightening and above 95 degrees C with little muscle birefringence. Injury adjacent to laser channels was greatest parallel to cell orientation. In vivo, muscle with reduced birefringence was surrounded by cells exhibiting focal birefringence increases (contraction bands). Early injury assessment marked by birefringence changes corresponded to lesion size at 3 days. CONCLUSION: Polarized light revealed histologic temperature signatures corresponding to irreversible muscle injury and collagen denaturation.

Animals↗

t(1) noise and sensitivity in pulsed field gradient experiments.

In this paper, a calculation routine based on product operator formalism and coherence pathway is presented, which describes t(1) noise and sensitivity in pulsed field gradient experiments. Several examples including the absolute value mode pulsed field gradient COSY, MQF-COSY, MQC, HMQC, and NOESY sequences are investigated to study the t(1) noise stemming from the phase errors of radiofrequency (RF) pulses and the sensitivity affected by RF pulse rotation angles. Our theoretical results indicate that the t(1) noises in P-type COSY, multiple-quantum-filtered (MQF)-COSY, and multiple-quantum coherence (MQC) are lower than those in the N-type corresponding experiments, while in HMQC and NOESY there is no difference in t(1) noise effects between P-type and N-type spectra. Meanwhile, from the calculations, we obtained the optimized RF pulse rotation angles in those sequences. In MQF-COSY, an increase in sensitivity of about 4(cos(2)&theta; sin(q-1)2&theta;)(2) - 1 (&theta; = arc cot can be achieved by using the optimized angles. In MQC, the increase is 2 cos(2)&theta; sin(q-1)2&theta; - 1 (&theta; = arc cot. MQF-COSY experiments are also carried out to support our corresponding theoretical results.

Magnetic Resonance Spectroscopy↗

Photocatalytic degradation of AZO dyes by supported TiO2 + UV in aqueous solution.

The photocatalytic degradation performance of photocatalysts TiO2 supported on 13-X, Na-Y, 4A zeolites with different loading content was evaluated using the photocatalytic oxidation of dyes direct fast scarlet 4BS and acid red 3B in aqueous medium. The results showed that the best reaction dosage of TiO2-zeolite catalysts is about 2 g/l and the photocatalytic kinetics follows first order for all supported catalysts. The photocatalytic activity order of the three series catalysts is 13X type >Y type >4A type. The physical state of titanium dioxide on the supports is evaluated by X-ray photoelectron spectra (XPS), powder X-ray diffraction (XRD), BET, and FTIR.

Azo Compounds↗

Development and validation of a calcium intake questionnaire for postmenopausal women in China.

PURPOSE: Develop and evaluate a food frequency questionnaire (FFQ) for assessing calcium intake in Chinese postmenopausal women. METHODS: An interviewer-conducted FFQ that asks respondents about their consumption of 110 calcium-containing foods over the previous week was developed and administered to 21 randomly selected postmenopausal women from Chengdu. This FFQ uses both the traditional weight-estimation method of assessing portion size in China and a new volume-estimation method. Test-retest reliability was assessed by re-administering the questionnaire two weeks later and validity was assessed by comparing the computed daily calcium intake to that obtained by a researcher-conducted 4-day food record. RESULTS: Estimates of calcium intake derived from the volume-estimation method were significantly lower than estimates derived from the traditional weight-estimation method (median calcium intakes = 270 mg/day versus 570 mg/day), but were quite similar to those obtained from the 4-day food record (median calcium intake = 275 mg/day). The test-retest reliability and validity of the volume-estimation method (intraclass correlation coefficient (ICC) = 0.73 and 0.86, respectively) were better than those of the weight-estimation method (ICC = 0.58 and 0.39, respectively). CONCLUSIONS: The reliability and validity of the traditional weight-estimation FFQ method of assessing dietary intake of nutrients in China is poor; this method may significantly overestimate dietary intakes. The proposed volume-estimation FFQ method for assessing calcium intake is a simple, interviewer-conducted method that is both reliable and valid. The calcium intake of postmenopausal women in urban China is much lower than the recommended daily allowance of 800 mg.

Aged↗

Loss of factor VIII activity during storage in PVC containers due to adsorption.

Recombinant factor VIII concentrates are stable when administered in a reconstituted form according to the manufacturer's specifications, and undiluted via infusion with syringe mini-pumps. However many Haemophilia centres administer recombinant factor VIII further diluted in intravenous fluids for greater ease of administration. To investigate the stability of recombinant factor VIII during administration as a diluted infusion, reconstituted factor VIII was stored in polyvinylchloride (PVC) mini-bags undiluted (146 IU mL-1) and at factor VIII concentrations of 10 IU mL-1 and 2 IU mL-1. After 48 h of storage at room temperature in PVC mini-bags, the recoveries of factor VIII activity were 41.9% of the initial activity for the undiluted (146 IU mL-1) product and 43.7% of the initial activity for factor VIII diluted to 10 IU mL-1. For factor VIII diluted to 2 IU mL-1, the amount of factor VIII activity remaining at 48 h was only 1.8% of the initial activity. In contrast, 100% of factor VIII activity was recovered after 48 h when undiluted reconstituted product (146 IU mL-1) was stored in a syringe. To investigate the mechanism of factor VIII activity loss during storage, factor VIII samples collected after 0, 3 and 48 h of storage were analysed by immunoblotting with factor VIII antibodies. No evidence of factor VIII proteolytic degradation during storage was found, however, large amounts of factor VIII antigen were recovered from the empty PVC mini-bags following elution with denaturing detergent. We conclude that clinically significant losses of factor VIII activity occur during storage in PVC mini-bags and that the loss of activity is most likely due to protein adsorption onto the plastic surface. This loss of factor VIII activity during storage in PVC containers may substantially affect the safety and potential cost savings of administering recombinant factor VIII by continuous infusion.

Adsorption↗

Two new acyclic diterpene-gamma-lactones from Salix matsudan.

Two new phytane-type diterpene-gamma-lactones named hanliuine I (1) and hanliuine II (2), were isolated from leaves of Salix matsudana (chinese name "hanliu", Berberidaceae). Their structures were deduced from 1D and 2D NMR techniques.

4-Butyrolactone↗

NAD(P)H:quinone oxidoreductase-dependent risk for colorectal cancer and its association with the presence of K-ras mutations in tumors.

NAD(P)H:quinone oxidoreductase (NQO1) is a polymorphic enzyme involved in the detoxification of potentially mutagenic and carcinogenic quinones. The homozygous C609T NQO1 genotype resulting in loss of reductase activity is found in 2-20% of individuals. In the present study, the NQO1-dependent risk for sporadic colorectal cancer (CRC) was studied in 247 incident CRC cases and 296 hospital-based controls recruited during 1996-1997. Four subgroups of cases were studied: (i) all CRCs; (ii) a molecular CRC subgroup (n = 117, cases with molecular tumor analyses); (iii) within the molecular subgroup those tumors with K-ras mutations in codon 12 (CRC K12); (iv) within the molecular subgroup those tumors with K-ras mutations in codon 13 (CRC K13). The C609T NQO1 genotype was found to be twice as prevalent in all CRC patients (6.8%) compared with controls (3%) and six times more common in the subset CRC K12 (20%). Multivariant analyses in the overall population of 247 cases and 296 controls showed a significant age and gender adjusted risk for CRC associated with the C609T NQO1 genotype (OR 2.9, 95% CI 1.19-6.97; P = 0.01) or with any variant genotype (the low activity allele frequency, i.e. heterozygotes plus homozygotes) (OR 1.41, 95% CI 1.02-1.92; P = 0.03). Within cases of the molecular subgroup (n = 117) the C609T NQO1 genotype was associated with the presence of K-ras codon 12 mutation (OR 6.5 95%, CI 1.39-34.9; P = 0.003). Logistic regression showed an age and gender adjusted risk for K-ras codon 12 mutant CRC associated with the C609T NQO1 genotype (OR 10.5, 95% CI 2.99-36.7; P: = 0.0002) or with any variant NQO1 genotype (OR 2.23, 95% CI 1.23-4.00; P = 0.007) compared with the control group. Genetically determined variations in NQO1 may modify the risk for CRC and these risks may be greatest for tumors containing K-ras codon 12 mutations. CRC with K-ras codon 12 mutations may represent a distinct and etiologically more homogeneous subtype of the disease, which may be associated with toxicants that are metabolized via a NQO1-dependent pathway.

Age Factors↗

Correlations of partial and extensive methylation at the p14(ARF) locus with reduced mRNA expression in colorectal cancer cell lines and clinicopathological features in primary tumors.

p14(ARF) is a putative tumor suppressor gene thought to modify the levels of p53. CpG sites within the 5'-flanking region and exon 1beta of p14(ARF) are targets of aberrant methylation and transcriptional silencing in human colorectal cancer (CRC). Here we have developed methylation-specific polymerase chain reaction (MSPCR) methods to detect methylation of CpG sites in p14(ARF) in CRC cell lines and primary CRC tumors, and correlated p14(ARF) mRNA expression with methylation in CRC cell lines using competitive quantitative reverse transcription-polymerase chain reaction methods. Ten CRC cell lines were studied; three (DLD-1, HCT15 and SW48) showed extensive methylation and six (Colo320, SW480, HT29, Caco2, SW837 and WiDr) were unmethylated; the other cell line, LoVo, showed partial methylation that affected exon 1beta but not the immediate upstream CpG sites. p14(ARF) mRNA was expressed at extremely low levels in fully methylated cell lines and at 10(4)- to 10(5)-fold higher levels in unmethylated cell lines. p14(ARF) expression in the partially methylated LoVo cell line was intermediate. Treatment of LoVo cells with 2 microM 5-aza-2'-deoxycytidine for 72 h was associated with marked (100-fold) induction of mRNA levels. Of 119 primary CRCs, 18% contained p14(ARF) methylation, although partial methylation was the most common pattern observed (in 67% of methylated tumors). Methylation of p14(ARF) was often accompanied by p16(INK4a) methylation; however, 50% of p14(ARF) methylated tumors contained unmethylated p16(INK4a). Methylation at p14(ARF) was associated with female gender, greater age, proximal anatomic location and poor differentiation, but not stage at diagnosis. A two-step MSPCR method for assaying p14(ARF) methylation in human tumors is described.

Adult↗

Phylogeny of Vibrio cholerae based on recA sequence.

We sequenced a 705-bp fragment of the recA gene from 113 Vibrio cholerae strains and closely related species. One hundred eighty-seven nucleotides were phylogenetically informative, 55 were phylogenetically uninformative, and 463 were invariant. Not unexpectedly, Vibrio parahaemolyticus and Vibrio vulnificus strains formed out-groups; we also identified isolates which resembled V. cholerae biochemically but which did not cluster with V. cholerae. In many instances, V. cholerae serogroup designations did not correlate with phylogeny, as reflected by recA sequence divergence. This observation is consistent with the idea that there is horizontal transfer of O-antigen biosynthesis genes among V. cholerae strains.

Base Sequence↗

Lymphatic absorption of fatty acids and cholesterol in the neonatal rat.

High-fat diets are essential in suckling animals to ensure adequate calories for postnatal growth, but their lymphatic transport of dietary lipids has not been characterized. We established a lymph fistula model in suckling rats to quantify intestinal uptake and lymphatic transport of dietary lipids and analyzed lipoprotein fractions. Suckling 19-day-old Sprague-Dawley rats had their mesenteric lymph ducts cannulated and gastroduodenal tubes inserted. After overnight recovery, [(3)H]triolein and [(14)C]cholesterol were infused for 6 h. Of the total dose, only 38% of triolein and 24% of cholesterol were transported in the lymph of suckling rats. Analyses of residual luminal contents and intestinal mucosal homogenate showed neither reduced absorption nor delayed mucosal processing of ingested lipids to be the cause. Thin-layer chromatographic analysis of radioactive mucosal lipids, however, showed a predominance of free fatty acids (60%) and free cholesterol (67%), implying impaired esterification capacity in these animals. We speculate that this reduced esterification allows for portal transport or direct enterocyte metabolism of dietary lipids.

Age Factors↗

Adenovirus-mediated human pancreatic lipase gene transfer to rat bile: gene therapy of fat malabsorption.

This study explored the potential of using the gene therapy approach, based on adenovirus-mediated expression of pancreatic lipase in the hepatobiliary tract, to increase lipid digestion in the intestinal lumen and promote lipid absorption through the gastrointestinal tract. Recombinant adenovirus containing the human pancreatic lipase cDNA (AdPL) was shown to transduce and mediate pancreatic lipase biosynthesis in rat IEC-6 epithelial cells in vitro. Retrograde infusion of recombinant adenovirus (3 x 10(8) plaque-forming units) containing the bacterial LacZ gene (AdLacZ) into the bile duct of rats resulted in positive X-gal reaction products in the periportal liver cells 7 days after AdLacZ infusion. A high level of human pancreatic lipase was detected in bile after retrograde bile duct infusion of rats with AdPL but not in the bile of animals infused with AdLacZ. Triglyceride hydrolytic activity in the bile of AdPL-infused rats was equivalent to that present in pancreatic juice. In contrast, serum obtained from these animals did not contain any detectable pancreatic lipase activity. These results suggest that ectopic expression of pancreatic enzymes in the hepatobiliary tract may be an alternative therapeutic strategy for treating fat malabsorption due to pancreatic insufficiency.

Adenoviridae↗