Search PubMed⌕ Search

Biomedical subjects

S Xu

Publications and source records attributed to S Xu.

At least 91 records · Page 5Linked to original sources

Production of cloned goats from enucleated oocytes injected with cumulus cell nuclei or fused with cumulus cells.

This study was designed to produce cloned goats from cumulus cells. Cloning donor nuclei were from cumulus cells either freshly isolated or cultured in vitro. Enucleated oocytes were either injected with cumulus cell nuclei without piezo-driven manipulator (injection method) or fused with cumulus cells (fusion method). The survival rate of cloned embryos, obtained by injection, was higher than that derived from fusion (62.7 and 45.9%, respectively). Two cloned goats were derived by fusion with in vitro cultured cumulus cells without starvation, but died shortly after natural birth, from respiratory difficulties. Their birth weights (2.23 kg and 2.03 kg) were within the normal range (2.0-2.7 kg) and postmortem analysis revealed no morphological abnormalities. The third cloned goat, derived by injection of nuclei from freshly isolated cumulus cells, weighed 3.3 kg at birth, and was 37% overweight compared with the average weight of the same species. This goat is healthy and well as this paper is being prepared. Nested PCR-RFLP analysis confirmed that all the cloned goats were derived from the donor cells.

Animals↗

Bayesian mapping of quantitative trait loci under complicated mating designs.

Quantitative trait loci (QTL) are easily studied in a biallelic system. Such a system requires the cross of two inbred lines presumably fixed for alternative alleles of the QTL. However, development of inbred lines can be time consuming and cost ineffective for species with long generation intervals and severe inbreeding depression. In addition, restriction of the investigation to a biallelic system can sometimes be misleading because many potentially important allelic interactions do not have a chance to express and thus fail to be detected. A complicated mating design involving multiple alleles mimics the actual breeding system. However, it is difficult to develop the statistical model and algorithm using the classical maximum-likelihood method. In this study, we investigate the application of a Bayesian method implemented via the Markov chain Monte Carlo (MCMC) algorithm to QTL mapping under arbitrarily complicated mating designs. We develop the method under a mixed-model framework where the genetic values of founder alleles are treated as random and the nongenetic effects are treated as fixed. With the MCMC algorithm, we first draw the gene flows from the founders to the descendants for each QTL and then draw samples of the genetic parameters. Finally, we are able to simultaneously infer the posterior distribution of the number, the additive and dominance variances, and the chromosomal locations of all identified QTL.

Analysis of Variance↗

Isolation of Helicobacter cinaedi from the colon, liver, and mesenteric lymph node of a rhesus monkey with chronic colitis and hepatitis.

On the basis of biochemical, phenotypic, and 16S rRNA analyses, Helicobacter cinaedi was isolated from the colon, liver, and mesenteric lymph nodes of a 2-year-old rhesus monkey with chronic diarrhea. Histologically, the liver had mild to moderate biliary hyperplasia and hypertrophy with periportal inflammation and fibrosis. Colonic and cecal lesions consisted of diffuse chronic inflammation and glandular hyperplasia extending the length of the crypts. This is the first observation of H. cinaedi associated with active hepatitis and colitis in a nonhuman primate.

Animals↗

B-cell maturation protein, which binds the tumor necrosis factor family members BAFF and APRIL, is dispensable for humoral immune responses.

B-cell maturation protein (BCMA) is a member of the tumor necrosis factor (TNF) receptor family and is expressed in B lymphocytes. BCMA binds two TNF family members, BAFF and APRIL, that stimulate cellular proliferation. BAFF in particular has been shown to influence B-cell survival and activation, and transgenic mice overexpressing BAFF have a lupus-like autoimmune disorder. We have inactivated BCMA in the mouse germ line. BCMA(-/-) mice have normal B-cell development, and the life span of mutant B lymphocytes is comparable to that of wild-type B cells. The humoral immune responses of BCMA(-/-) mice to T-cell-independent antigens as well as high and low doses of T-cell-dependent antigens are also intact. In addition, mutant mice have normal splenic architecture, and germinal centers are formed during an ongoing immune response. These data suggest a functional redundancy of BCMA in B-cell physiology that is probably due to the presence of TACI, another TNF receptor family member that is expressed on B cells and that can also bind BAFF and APRIL.

Animals↗

Cell-specific basolateral membrane sorting of the human liver Na(+)-dependent bile acid cotransporter.

The human Na(+)-taurocholate cotransporting polypeptide (Ntcp) is located exclusively on the basolateral membrane of hepatocyte, but the mechanisms underlying its membrane sorting domain have not been fully elucidated. In the present study, a green fluorescent protein-fused human NTCP (NTCP-GFP) was constructed using the polymerase chain reaction and was stably transfected into Madin-Darby canine kidney (MDCK) and Caco-2 cells. Taurocholate uptake studies and confocal microscopy demonstrated that the polarity of basolateral surface expression of NTCP-GFP was maintained in MDCK cells but was lost in Caco-2 cells. Nocodazole (33 microM), an agent that causes microtubular depolymerization, partially disrupted the basolateral localization of NTCP-GFP by increasing apical surface expression to 33.5% compared with untreated cells (P < 0.05). Brefeldin A (BFA; 1-2 microM) disrupted the polarized basolateral localization of NTCP, but monensin (1.4 microM) had no affect on NTCP-GFP localization. In addition, low-temperature shift (20 degrees C) did not affect the polarized basolateral surface sorting of NTCP-GFP and repolarization of this protein after BFA interruption. In summary, these data suggest that the polarized basolateral localization of human NTCP is cell specific and is mediated by a novel sorting pathway that is BFA sensitive and monensin and low-temperature shift insensitive. The process may also involve microtubule motors.

Animals↗

The novel gene locus for agenesis of permanent teeth (He-Zhao deficiency) maps to chromosome 10q11.2.

He-Zhao deficiency has been recently characterized with a distinct form of agenesis of permanent teeth that is different from other previously reported disorders of tooth agenesis. This inherited abnormality suggests that some gene(s) associated with the development of permanent teeth may mutate. In this study, we map the gene locus to chromosome 10q11.2. The DNA pooling method combined with two-point and multi-point linkage analysis has been successfully applied. The maximum LOD (Zmax) scores for two-point and multi-point analyses are 13.29 (on marker D10S196) at recombination fraction (theta) = 0 and 18.09 (between markers D10S1772 and D10S1766), respectively. Haplotype analysis confined the locus within an interval of 5.5 cM flanked by markers D10S604 and D10S568. This study has demonstrated a novel gene locus responsible for He-Zhao deficiency and provides a good likelihood for the discovery of one of the genes determining permanent tooth formation and development.

Alleles↗

Cholangiohepatitis and inflammatory bowel disease induced by a novel urease-negative Helicobacter species in A/J and Tac:ICR:HascidfRF mice.

Helicobacter bilis and H. hepaticus, both urease-positive intestinal helicobacters of mice, have been shown experimentally to induce proliferative typhlocolitis in scid mice. We recently isolated a urease-negative Helicobacter sp. (H. sp.) that also induced proliferative typhlocolitis in pilot studies in scid mice. To determine the pathogenic potential of H. sp. in immunocompromised and immunocompetent mice, 5-week old male A/J or Tac:Icr:Ha(ICR)-scidfRF mice were inoculated by intraperitoneal (IP) injection with approximately 3 x 10(7) colony-forming units (CFU) of H. sp. Mice were necropsied at various time points postinoculation (PI). Sham-inoculated mice had no clinical, gross, or histopathological lesions. In contrast, scid mice inoculated IP with H. sp. had severe hemorrhagic diarrhea and decreased weight gain at 2, 7, and 18 weeks postinoculation (PI), with severe proliferative typhlocolitis, phlebothrombosis, and hepatitis. A/J mice had no clinical signs, but had mild to moderate proliferative typhlocolitis and moderate to marked cholangiohepatitis at 7 and 24 weeks PI. A/J mice infected with H. sp. developed robust immune responses of a predominant Th1 type. This report demonstrates that infection with a urease-negative helicobacter can cause inflammatory bowel disease (IBD) and hepatitis in scid and immunocompetent A/J mice. These results provide a new model of IBD and cholangio-hepatitis associated with a specific urease-negative, novel H. species.

Animals↗

PPARalpha-dependent induction of liver microsomal esterification of estradiol and testosterone by a prototypical peroxisome proliferator.

Fatty acyl-coenzyme A:estradiol acyltransferase in liver microsomes catalyzes the formation of estradiol fatty acid esters. These estrogen esters are extremely lipophilic and have prolonged hormonal activity because they are slowly metabolized and slowly release estradiol. Our previous studies showed that treatment of female rats with clofibrate or gemfibrozil (peroxisome proliferators commonly used as hypolipidemic drugs) markedly stimulated the liver microsomal esterification of estradiol. Although clofibrate administration is a potent inducer of liver microsomal fatty acyl-coenzyme A:estradiol acyltransferase in rats, it is a poor inducer in mice. In contrast to these observations, Wy-14,643 (an exceptionally potent prototypical peroxisome proliferator) is a strong inducer of fatty acyl-coenzyme A:estradiol acyltransferase in mice. To explore the role of PPARalpha in the induction of fatty acyl-coenzyme A:estradiol acyltransferase and fatty acyl-coenzyme A:testosterone acyltransferase activities by peroxisome proliferators, we fed 0.1% Wy-14,643 to female wild-type and PPARalpha null mice for 11 d. The liver microsomal acyl-coenzyme A:estradiol acyltransferase and acyl-coenzyme A:testosterone acyltransferase activities were increased 4- to 5-fold in wild-type mice fed Wy-14,643, but no increase was observed in null mice. These results demonstrate that induction of acyl-coenzyme A:estradiol acyltransferase and acyl-coenzyme A:testosterone acyltransferase activities by a prototypical peroxisome proliferator is dependent on PPARalpha.

Acyltransferases↗

Immunomodulating activity of CVT-E002, a proprietary extract from North American ginseng (Panax quinquefolium).

The activity of CVT-E002, an aqueous extract containing mainly oligosaccharides and polysaccharides from North American ginseng (Panax quinquefolium), as an immunobooster on murine spleen cells and peritoneal macrophages, was studied in-vitro. CVT-E002 stimulated the proliferation of normal mouse spleen cells, of which the major responding subpopulation was identified as B lymphocytes. CVT-E002 also activated peritoneal exudate macrophages leading to enhanced interleukin-1 (IL-1), interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) production. In addition, CVT-E002 stimulated in-vivo immunoglobulin G (IgG) production in treated mice. These results identify some of the immunomodulating activities of CVT-E002 and suggest its use clinically for the modulation of immune responses.

Adjuvants, Immunologic↗

Photobleaching of hypocrellin B and its butylamino-substituted derivative in solutions.

The photobleaching of hypocrellin B (HB) and its derivative butylamino-substituted hypocrellin B (BAHB), both of which are potent sensitizers for photodynamic therapy (PDT), were investigated by studies of absorption spectra and quenching experiments and by the determination of photoproducts. Control experiments indicated that the sensitizer, oxygen and light were essential for the photobleaching of HB and BAHB, which suggested that it was a photodynamic process, e.g. the photobleaching processes of both HB and BAHB were mainly self-sensitized photooxidations. The illumination of HB with visible light in oxygenated nonpolar solvents generated singlet oxygen efficiently [phi(1O2) = 0.76] which in turn attacked the sensitizer HB with the subsequent formation of an endoperoxide product. The endoperoxide of HB is unstable at room temperature and undergoes loss of singlet oxygen with regeneration of the parent HB. The singlet oxygen released from the endoperoxide of HB was detected with chemical trapping experiments. The quenching experiments indicated that in increasingly polar solvents the superoxide anion mechanism (type I) as well as the singlet oxygen mechanism (type II) contributed to the photobleaching of HB. The introduction of the electron-donating butylamino group not only enhanced the yield of the superoxide anion generation but also altered the position of attack in the BAHB molecule by the activated oxygen species. No endoperoxide product was observed, and no singlet oxygen released from the photobleaching process of BAHB was detected. The photobleaching process of BAHB was more complex. Both the singlet oxygen and superoxide anion mechanism played important roles in the photobleaching of BAHB in all organic solvent used here, even in aerobic nonpolar solvents such as CHCl3.

Electron Transport↗

Stimulatory effect of clofibrate and gemfibrozil administration on the formation of fatty acid esters of estradiol by rat liver microsomes.

Fatty acyl-coenzyme A (CoA):estradiol acyltransferase in liver microsomes catalyzes the formation of estradiol fatty acid esters. These esters are lipophilic and have prolonged hormonal activity because they are slowly metabolized and because they slowly release estradiol. In the present study, we have shown that treatment of rats with clofibrate or gemfibrozil (peroxisome proliferators that are commonly used hypolipidemic drugs) markedly stimulate the liver microsomal esterification of estradiol. Administration of 0.15, 0.30, 0.45, or 0.60% clofibrate in an AIN-76A diet to female rats for 4 weeks stimulated fatty acyl-CoA:estradiol acyltransferase activity per milligram of microsomal protein by 4-, 8-, 14- and 16-fold, respectively, when estradiol was incubated with liver microsomes and a fatty acyl-CoA. Additional studies showed that incubation of (3)H-labeled estradiol with liver microsomes, ATP, and coenzyme A resulted in the formation of multiple fatty acid esters of estradiol from endogenous fatty acids in liver microsomes, and the formation of these esters was stimulated manyfold by pretreatment of rats with clofibrate. This study provides the first demonstration of a stimulatory effect of an environmental agent on the esterification of an estrogen.

Acyltransferases↗

Stimulatory effect of clofibrate on the action of estradiol in the mammary gland but not in the uterus of rats.

Treatment of ovariectomized rats with dietary clofibrate caused a manyfold increase in the liver microsomal esterification of estradiol with fatty acids. The stimulatory effect of clofibrate administration on fatty acid esterification of estradiol by liver microsomes was paralleled in vivo by enhanced estradiol-induced increases in the formation of lobules in the mammary gland and by increased incorporation of bromodeoxyuridine into these lobules. In contrast to the stimulatory effect of clofibrate administration on the action of estradiol in the mammary gland, clofibrate administration had no effect on the uterotropic action of estradiol. These results indicate that clofibrate administration has a selective stimulatory effect on the hormonal action of estradiol in the mammary gland but not in the uterus.

Acyl Coenzyme A↗

Tumor-suppressive effects of neutral endopeptidase in androgen-independent prostate cancer cells.

Expression of neutral endopeptidase (NEP) 24.11 is diminished in metastatic, androgen-independent prostate cancers (PCs; C. N. Papandreou et al., NAT: MED:, 4: 50--57, 1998). To determine the effects on androgen-independent PC cells of overexpressing cell-surface NEP, an inducible tetracycline-regulatory gene expression system was used to stably introduce and express the NEP gene in androgen-independent TSU-Pr1 cells generating WT-5 cells, which expressed high levels of enzymatically active NEP protein when cultured in the absence of tetracycline. TN12 cells, which contain the identical vectors without the NEP gene and do not express NEP, were used as control. Expression of NEP in WT-5 cells after removal of tetracycline from the media resulted in a >80% inhibition in cell proliferation over a 1-week period (P < 0.005) compared with control cells. Tumor formation occurred in the prostate glands of orthotopically injected athymic mice killed at 30 days in 4 of 5 mice that were given injections of 2 x 10(6) WT-5 cells and were fed doxycycline (NEP suppressed), and in all mice that were given injections of TN12 cells and were fed with or without doxycycline. In contrast, only 1 of 5 mouse prostates developed a tumor in mice that were given injections of WT-5 cells and that did not receive doxycycline. Analysis of the mechanisms of NEP-induced growth suppression revealed that NEP expression in WT-5 cells induced a 4-fold increase in the number of PC cells undergoing apoptosis, and increased the expression of p21 tumor suppressor gene protein and the level of unphosphorylated retinoblastoma protein as determined by Western blot. Flow cytometric analysis show that induced NEP expression in WT-5 cells resulted in a G(1) cell cycle arrest. These data show that NEP can inhibit PC cell growth and tumorigenicity and suggest that NEP has potential as therapy for androgen-independent PC.

Androgens↗

[Modification of the Photobacterium phosphoreum toxicity test method].

In order to reduce the variation of Photobacterium phosphoreum luminous intensity during testing, the Photobacterium phosphoreum toxicity test was modified by a correction factor, PIacetone, in this paper. The effects of magnetic stirring time, culture time, culture generation and culture temperature on the measured results were discussed. The modified method had good laboratory repeatability and the relative standard deviation (RSD) was between 2.1% and 13.1%. Reliable data could be obtained using the modified method. It would be beneficial to the development of the quantitative structure-activity relationships on multicomponent organic compounds.

Luminescent Measurements↗

[Preliminary analysis of genetic characteristics of the nonstructural protein NSP4 of rotavirus strains with different serotypes in China].

OBJECTIVE: To study the genotypes and genetic variations of nonstructural protein NSP4 of rotavirus strains with different serotypes in China. METHODS: NSP4 cDNA of the four serotypes G1, G2, G3 and G9 were cloned and sequenced by SILVER SEQUENCE DNA Sequencing System. RESULTS: The identity of the nucleotide and amino acid were 78.8%-93.5%, 83.0%-95.5%, respectively. NSP4 from strains G1, G3, G9 all fell into Wa allele, while that of G9 fell into KUN allele. Some variations of the amino acids were observed localizing at the position of 135-145 in the VP4 binding domain. CONCLUSION: The results indicated that the two alleles Wa and KUN are the major prevalent NSP4 genotypes in China.

Animals↗

[The genotype analysis of glucose-6-phosphate dehydrogenase deficiency in Yunnan province].

OBJECTIVE: To identify glucose-6-phosphate dehydrogenase (G6PD) gene mutations in 23 patients with G6PD deficiency and to gain further understanding of the molecular and genetic background of G6PD gene in Yunnan province, China. METHODS: The mutations located in exons 2-12 and in parts of introns of G6PD gene were analyzed by amplification refractory mutation system(ARMS), natural and mis-match primer PCR/restrict enzyme, polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP ) analysis and automatic DNA sequencing. RESULTS: Among these 23 samples, 5 different point mutations in G6PD gene were identified, and they constituted 5 genotypes. There were 7 Han and 3 Dai patients with G487A mutation, 7 cases with both intron 11 T93C and C1311T mutations, 4 cases with intron 5 636 or 637 T-->del mutation, 1 case with G871A mutation, and 1 case with G487A/T93C/C1311T mutation. Two haplotypes, 93C/1311T and 93C/1311T/487A were identified in Yunnan. A strong association was observed between C1311T and the Nla III restriction site produced by intron 11 T93C. The findings of the investigators on IVS-5 636 or 637T-->del in Chinese, on G871A in mainland of China, and on G487A in the Han people of Yunnan have not been reported previously. CONCLUSION: G6PD deficiency is very heterogenous in Yunnan; G487A is one of the common mutations in that province and may be of different origins. Possibly IVS-11 T93C mutation is of non-African origin. IVS-11 T93C and C1311T might jointly result in G6PD deficiency. The above data on G6PD gene mutation types could be useful for clinical diagnosis, prevention of G6PD deficiency, and researches in the origin and migration of minorities in Yunnan or other regions.

Adolescent↗