Differential role of the vagus in diurnal gene expression rhythms in the small intestine

The objective of this study was to examine the function of vagal innervation in maintaining diurnal rhythmicity in the expression of intestinal absorptive genes. Rats underwent truncal vagotomy and were maintained for 7 days on nighttime scheduled feeding (12-h light/12-h dark cycle). Vagotomized rats (V; n = 9) were pair-fed with sham-operated controls (S; n = 4). Unoperated normal rats (N; n = 6) were also included as controls. Half the rats were killed 3 h after lights on (ZT3; Zeitgeber Time, with lights-on considered ZT0) and the other half at ZT9, the time interval over which we have previously shown that sucrase and sugar transporter expression exhibits a significant anticipatory increase. RNA and protein extracted from mucosa of proximal jejunums were subjected to Northern and Western blot analyses to assess the increase in gene expression. Sham operation did not alter the normal diurnal rhythmicity of intestinal gene expression. Control rats (S plus N) exhibited the expected increase in RNA levels at ZT9 versus ZT3 for SGLT1 (4.5-fold), GLUT2 (5.3-fold), GLUT5 (4.1-fold), and sucrase (2.9-fold; P > 0.001 in all cases). In contrast, the induction in V rats was markedly blunted for GLUT2 (1.3-fold) and sucrase (1.5-fold) but not for SGLT1 (5.0-fold) or GLUT5 (4.2-fold). The mRNA levels for GLUT2 and sucrase at ZT9 were significantly lower in V rats versus controls (P < 0.001). GLUT2 and SGLT1 protein levels exhibited a parallel pattern: SGLT1 induction was 4.3-fold in control rats (P < 0.01) and 3.8-fold in V rats (P <0.01), whereas GLUT2 induction was 3.3-fold in control rats (P < 0.01) but only 1.4-fold in V rats (NS). Our results indicate that signaling through the vagus nerve is necessary to maintain the anticipatory induction pattern of GLUT2 and sucrase. The persistent rhythm in both SGLT1 and GLUT5 indicates that (1) diurnal induction of these genes is independent of vagal innervation and (2) the procedure did not cause an overall loss of intestinal function. Thus, entrainment of anticipatory diurnal gene expression in the intestine occurs via two separate pathways that are differentially dependent on vagal input.     

无环鸟苷亲脂性前体药物脂质体的制备及体外抗病毒活性(英文)

本文通过将无环鸟苷(acyclovir,简称ACV)2’位羟基分别与月桂酰氯或棕榈酰氯进行酯化反应,制得亲脂性前体药物无环鸟苷月桂酸酯和无环鸟苷棕榈酸酯(分别简称为C₁₂-ACV和C₁₆-ACV),使脂质体包封率从ACV的29.9%提高到C₁₂-ACV的95.6%和C₁₆-ACV的97.1%;漏泄实验表明在4℃透析60h后,一半以上的ACV从脂质体中漏泄,而C₁₂-ACV和C₁₆-ACV的滞留率分别为70%和80%;体外抗疱疹病毒的试验中,在最低试验浓度0.044μmol/L时,ACV不显示抗病毒活性,而C₁₆-ACV脂质体抑制细胞病变率达75%,说明前体药物通过与脂质体脂膜的结合增加了药物的进入细胞能力,从而提高了ACV的抗病毒能力。     

Refractory potassium repletion due to cisplatin-induced magnesium depletion

Cisplatin is a common cause of hypomagnesemia and hypokalemia due to renal magnesium (Mg) and potassium (K) losses. Magnesium plays an important role in the maintenance of intracellular K. An unrecognized and untreated Mg depletion can lead to a refractory K repletion. We describe two patients with hypomagnesemia-associated refractory hypokalemia following cisplatin following cisplatin therapy. Potassium supplementation failed to replace the K deficit. Profound hypokalemia persisted until hypomagnesemia was recognized and corrected. In neither patient was the concurrent hypomagnesemia recognized until the 11th and 9th hospital days. These two cases demonstrated the association of a refractory K repletion and an Mg deficiency. Thus, both serum K ion and Mg levels should routinely be assessed in patients who require cisplatin therapy.     

Characterization of iridium film as a stimulating neural electrode

Iridium films having near-bulk properties were formed by electron-beam evaporation with simultaneous bombardment of Ar ion beam. The charge-injection capabilities of Ir film were investigated, and the detrusor pressure induced by S2 stimulation with Ir-coated Pt electrode was measured and compared with the uncoated Pt electrode. The charge densities of Ir film were continuously increased with increase in the number of cycles in 0.1 M H2SO4 due to the accumulation of the iridium oxide phase. The iridium oxide formed contained nano-pores, and oxides had different dielectric properties. The Ir film could inject various amounts of charge in physiological solution under the identical stimulating condition depending on the degree of activation in 0.1 M H2SO4. S2 stimulation by Ir-coated Pt electrode caused more efficient bladder contraction of the male dog than the uncoated Pt electrode under the identical stimulus condition.     

Development of hatching blastocysts from immature human oocytes following in-vitro maturation and fertilization using a co-culture system

Recently, in-vitro maturation (IVM) of immature human oocytes recovered from non-stimulated follicles has been applied in the treatment of infertility. However, in previous reports, very few embryos cultured in conventional medium have reached the expanded blastocyst stage following in-vitro maturation and fertilization (IVM/IVF). The objective of this study was to investigate whether the developmental competence of human embryos following IVM/IVF could be enhanced by the use of a human ampullary cell co-culture system. Immature human oocytes were aspirated from small follicles at Caesarean section and then cultured in medium containing human menopausal gonadotrophin for 36 to 48 h, followed by insemination. Zygotes were randomly cultured either in conventional culture medium alone or in the co-culture system. Of 48 embryos cultured in conventional medium alone, all arrested at the 2-16- cell stage on day 3 after insemination. Of 46 embryos cultured in the co-culture system, 26 embryos (56.5%) arrested at the 2-16-cell stage. Six embryos (13%) developed to the morula stage. Fourteen embryos (30.4%) developed to expanded blastocysts and two blastocysts were hatching on day 7 after insemination. We conclude that co-culture significantly enhances the development of blastocysts in embryos resulting from IVM/IVF.      

Differences in antibody response of rabbit to intravenously injected soluble and cell-attached enterobacterial antigen

A study was made of the antigenicity of soluble and cell-attached common antigen obtained from enteric bacteria. This antigen becomes readily attached to erythrocytes, which become agglutinable in the presence of the corresponding antibody. Although the common antigen is extractable from numerous species and serogroups of Enterobacteriaceae, in only a few, notably Escherichia coli O14, does it engender antibodies in the rabbit upon intravenous injection. It is shown that antibodies against the common antigen are produced after intravenous injection of cell-attached antigen from E. coli O111, Salmonella spp. and Shigella spp. but not, or only slightly, after administration of soluble antigen. Antibodies are also formed after injection of antigen attached to C3H mouse fibroblast (L) cells. Intravenous injection of equivalent amounts of common antigen from E. coli O14, E. coli O111, S. typhi-murium and S. flexneri results in antibody formation against the common antigen only with that from E. coli O14.     

HLA-A*02 allele frequencies and haplotypic associations in Koreans

We have investigated the frequencies of HLA-A*02 alleles and their haplotypic associations with HLA-B and -DRB1 loci in 439 healthy unrelated Koreans, including 214 parents from 107 families. All of the 227 samples (51.7%) typed as A2 by serology were analyzed for A*02 alleles using polymerase chain reaction (PCR)-low ionic strength-single-strand conformation polymorphism (LIS-SSCP) method. A total of six different A*02 alleles were detected (A*02 allele frequency 29.6%): A*0201/9 (16.6%), *0203 (0.5%), *0206 (9.3%), *0207 (3.0%), and one each case of *0210 and *02 undetermined type. Two characteristic haplotypes showing the strongest linkage disequilibrium were A*0203-B38-DRB]*1502 and A*0207-B46-DRB1*0803. Besides these strong associations, significant two-locus associations (P<0.001) were observed for A*0201 with B61, DRB1*0901 and DRB1*1401, and for A*0206 with B48 and B61. HLA haplotypes carrying HLA-A2 showed a variable distribution of A*02 alleles, and all of the eight most common A2-B-DR haplotypes occurring at frequencies of > or =1% were variably associated with two different A*02 alleles. These results demonstrate that substantial heterogeneity is present in the distribution of HLA-A*02 alleles and related haplotypes in Koreans.     

HLA-B*07 allele frequencies and haplotypic associations in Koreans

We have investigated the frequencies of HLA-B*07 alleles and their haplotypic associations with HLA-A, -C and -DRB1 loci in 489 healthy unrelated Koreans, including 214 parents from 107 families. All of the 45 samples (9.2%) typed as B7 by serology were analyzed for B*07 alleles using polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) method. Two different B*07 alleles were detected: B*0702 (allele frequency 0.041) and B*0705 (0.005). Two characteristic haplotypes showing strong linkage disequilibrium in Koreans were A*2402-Cw*07-B*0702-DRB1*0101 (haplotype frequency 0.028) and A*2901-B*0705-DRB1*0803 (0.005). The characteristic haplotype A*2901-B*0705-DRB1*0803, found in 100% (5/5) of B*0705-positive individuals, has not been previously described in other ethnic groups. HLA-B7 alleles comprise distinctive extended haplotypes in the Korean population. The probability of HLA-B7 allele mismatches among ABDR-matched unrelated donor-recipient pairs is expected to be low in Koreans.     

Relationship between HLA-DRB1 and DQ alleles and the genetic susceptibility to type 1 diabetes

Objective　To study the relationship between human leukocyte antigen (HLA)-DRB1 and DQ alleles and the genetic susceptibility of type 1 diabetes in North Chinese children. Methods　Polymerase chain reaction (PCR) techniques were used to amplify the second exon of DRB1 and DQ alleles, after which sequence specific olignucleotide probe (SSOP) dot blot hybridization techniques were used to analyze the amplified products. Results　DRB1*0301, DQA1*0301, DQB1*0201 alleles and DRB1*0301-DQA1*0501-DQB1*0201 haplotype were significantly increased in patients, while DQA1*0103 and DQB1*0601 alleles were significantly increased in controls. The distribution of DR4 and DR9 haplotypes in patients and controls were not significantly different, but DR3/DR4 and DR4/DR9 heterozygotes were significantly increased in patients. Conclusions　DRB1*0301, DQA1*0301 and DQB1*0201 confer susceptibility while DQA1*0103 and DQB1*0601 confer protection to type 1 diabetes. DRB1*0301-DQA1*0501-DQB1*0201 haplotype offers a predisposition to type 1 diabetes in North Chinese. Although the distribution of DR4 and DR9 in patients and controls had no significant difference, DR3/DR4 and DR3/DR9 heterozygotes were significantly increased in patients, showing that the susceptive effects of DR3 and DR4 or DR4 and DR9 haplotypes could be added up.