Correlation analysis of MR/CT on colorectal cancer lymph node metastasis characteristics and prognosis.

OBJECTIVE: Colorectal cancer incidence showed increasing trend with living standard improvement and lifestyle changes. This study investigated the relationship and diagnostic value of colorectal cancer lymph node metastasis and prognosis using magnetic resonance (MR) and computed tomography (CT). PATIENTS AND METHODS: Siemens Avanto 3.0T MR and GE Light Speed Pro 64 row helical CT were used to scan 318 cases of colorectal cancer patients diagnosed by pathology. The relationship between colorectal cancer lymph node metastasis and prognosis after surgery was analyzed. RESULTS: The accuracy of MR and CT in judging colorectal cancer lymph node metastasis ratio (LNR) was 92.5% and 75.5%, respectively. MR showed significantly higher accuracy than CT (p < 0.05). The coincidence rate of LNR result derived from MR and CT with colorectal cancer histopathological results was 57.6% and 54.7%, respectively. MR and CT sensitivity were 42.6% and 25.0%, while their specificity was 74.1% and 41.3%, respectively. The positive predictive value and negative predictive value of MR and CT were 61.1% and 51.4%, 57.1% and 66.7%, respectively. c2-test showed that MR diagnosis result was consistent with histopathological result (p < 0.05). The coincidence rate of MR and CT evaluation on 5-year disease-free survival and overall survival were 56.7% and 43.8%, respectively. CONCLUSIONS: MR showed a better effect on prognosis than CT and could be treated as the first choice to predict LNR and prognosis. MR demonstrated a good correlation with pathological results and could be used to predict LNR and prognosis.

Dietary spray-dried chicken plasma improves intestinal barrier function and modulates immune status in weaning piglets.

The objective of this study was to evaluate the effects of dietary addition of spray-dried chicken plasma (SDCP) as a replacement for spray-dried porcine plasma (SDPP) on serum biochemistry, intestinal barrier function, immune parameters, and the expression of intestinal development-related genes in weaning pigs. One hundred and forty-four 25-d-old weaning piglets with BW of 6.43 ± 0.39 kg were randomly allotted to 1 of 4 dietary treatments: 1) CON (basal diet; control), 2) SDPP (containing 5% SDPP), 3) SDPP + SDCP (containing 2.5% SDPP and 2.5% SDCP), and 4) SDCP (containing 5% SDCP). After a 28-d trial, 6 pigs from each treatment were randomly selected to collect serum and intestinal samples. On d 14 after the initiation of the trial, pigs in the SDPP, SDPP + SDCP, and SDCP groups had an increase ( < 0.05) in serum concentrations of total protein and IgG and a decrease ( < 0.05) in activities of alanine aminotransferase and diamine oxidase compared with the CON group. In the jejunum, supplementation with SDPP and SDCP reduced ( < 0.05) the concentration of tumor necrosis factor-α (TNF-α) and upregulated ( < 0.05) the mRNA levels of zonula occludens 1 (ZO-1), zonula occludens 2 (ZO-2), occludin (OCLN), Toll-like receptor 2 (TLR2), glucagon-like peptide 2 (GLP2), and IGF-1 compared with the CON group. In the ileum, feeding SDPP, SDPP + SDCP, and SDCP decreased ( < 0.05) the concentrations of TNF-α and secretory IgA (sIgA) and upregulated ( < 0.05) the mRNA levels of claudin 1 (CLDN-1) and TLR2 compared with feeding CON. However, there were no differences among the SDPP, SDPP + SDCP, and SDCP groups. Furthermore, supplementation with SDCP reduced ( < 0.05) the concentration of IL-10 and upregulated ( < 0.05) the mRNA levels of GLP-2, mucin 2 (MUC2), and trefoil factor family 3 (TFF3) in the ileum compared with feeding CON. Collectively, the current results indicate that dietary addition of SDCP has a beneficial influence on the health condition of weaning pigs by alleviating liver damage, promoting intestinal development, improving intestinal barrier function, and reducing overstimulation of immune response. The efficacy of SDCP is comparable to that of SDPP.

Spray-dried chicken plasma improves intestinal digestive function and regulates intestinal selected microflora in weaning piglets.

One hundred forty-four 25-d-old weaning piglets with BW of 6.43 ± 0.39 kg were used in a 28-d trail to evaluate the effects of dietary addition of spray-dried chicken plasma (SDCP) as a replacement for spray-dried porcine plasma (SDPP) on growth performance, nutrient digestibility, diarrhea incidence, small intestinal morphology, digestive enzyme activity, and microflora. Pigs were randomly allotted to 1 of 4 dietary treatments: 1) CON (control; a basal diet), 2) SDPP (containing 5% SDPP), 3) SDPP + SDCP (containing 2.5% SDPP and 2.5% SDCP), and 4) SDCP (containing 5% SDCP). Six pigs from each treatment were randomly selected to collect serum and intestinal samples. Compared with the CON group, both the SDPP and the SDPP + SDCP groups improved final BW of pigs (P < 0.05), but there were no differences among the SDPP, SDPP + SDCP, and SDCP groups. From d 1 to 14 and d 15 to 28, pigs fed the SDPP and SDPP + SDCP diets had a greater (P < 0.05) ADG than pigs fed the CON diet. During the overall period, both ADG and ADFI of pigs in the SDPP and SDPP + SDCP groups were improved (P < 0.05) compared with pigs in the CON group. Furthermore, pigs fed diets containing SDPP or SDCP had a greater (P < 0.05) apparent total tract digestibility (ATTD) of CP, ether extract, Ca, and ash and less (P < 0.05) incidence of diarrhea than pigs fed the CON diet. However, no differences were observed for ATTD and diarrhea incidence between the SDPP and SDCP groups. Compared with the CON group, duodenal villus height and the ratio of villi to crypt were increased (P < 0.05) in the SDPP, SDPP + SDCP, and SDCP groups and jejunal crypt depth was decreased in the SDPP + SDCP and SDCP groups (P < 0.05). Pigs in the SDPP group had greater (P < 0.05) activities of amylase, maltase, and trypsin than pigs in the CON group. However, no significant differences were observed between the SDCP and SDPP groups. Additionally, inclusion of SDCP in diet decreased (P < 0.05) the population of Escherichia coli. In conclusion, these results demonstrated that the addition of SDCP in pigs' diet had an effect similar to SDPP on improving growth performance through the promotion of the small intestinal development, increasing digestive enzymes activities, enhancing ATTD of nutrients, and decreasing diarrhea incidence.

Controllable porosity hydroxyapatite ceramics as spine cage: fabrication and properties evaluation.

A procedure was designed to prepare porosity-graded hydroxyapatite (HA) ceramics simulating the bimodal structure of natural bone, which could be used to build a cage that would promote the reconstruction of the anterior column after vertebrectomy or corpectomy in tumor and trauma surgery. HA ceramics with controllable pore size distribution and porosity were developed by using chitosan and Poly(vinyl alcohol) (PVA) as the pore-forming agents. HA ceramics with worthwhile properties such as a wide range of volume porosity (10-50%) and pore size (nanometer to 400 microm) can be obtained from this method, which allows the fabrication of HA ceramics with desirable porous characteristics simulating the bimodal natural bone architecture expected to provide advantages for bony fusion in the intervertebral foramina. When coated with chitosan-gelatin network, the bending strength of the porous HA ceramics significantly improved. The polymer network coated porous HA have potential application in the construction of cages for spinal operations.

The impact of differential binding of wild-type RARalpha, PML-, PLZF- and NPM-RARalpha fusion proteins towards transcriptional co-activator, RIP-140, on retinoic acid responses in acute promyelocytic leukemia.

Retinoic acid receptor (RA) heterodimer (RAR/RXR) activities have been shown to be repressed by transcriptional co-repressor, SMRT/N-CoR, in the absence of the ligand while upon all-trans retionic acid (ATRA) treatment, SMRT/N-CoR is dissociated from RARalpha leading to gene expression by the recruitment of transcriptional co-activators to the transcriptional complex. The difference in response to ATRA therapy between acute promyelocytic leukemia (APL) patients with PML-RARalpha fusion and PLZF-RARalpha fusion has recently been found to be partially due to the strong association of the transcriptional co-repressor, SMRT/N-CoR, with PLZF domain. We demonstrate that SMRT association, as with PML-RARalpha, can be released from NPM-RARalpha at pharmacological concentration of ATRA (10-6 M). Moreover, we show for the first time that the interaction between the transcriptional co-activator, RIP-140, and PML-, PLZF- or NPM-RARalpha fusion proteins can be positively stimulated by ATRA although they are less sensitive as compared with the wild-type RARalpha. Our results suggest that the dissociation of transcriptional co-repressors, SMRT/N-CoR, and recruitment of co-activators, eg RIP-140, to APL-associated fusion proteins constitute a common molecular mechanism in APL and underlie the responsiveness of the disease to RA therapy. Leukemia (2000) 14, 77-83.

Distinct leukemia phenotypes in transgenic mice and different corepressor interactions generated by promyelocytic leukemia variant fusion genes PLZF-RARalpha and NPM-RARalpha.

Acute promyelocytic leukemia (APL) is characterized by a specific chromosome translocation involving RARalpha and one of four fusion partners: PML, PLZF, NPM, and NuMA genes. To study the leukemogenic potential of the fusion genes in vivo, we generated transgenic mice with PLZF-RARalpha and NPM-RARalpha. PLZF-RARalpha transgenic animals developed chronic myeloid leukemia-like phenotypes at an early stage of life (within 3 months in five of six mice), whereas three NPM-RARalpha transgenic mice showed a spectrum of phenotypes from typical APL to chronic myeloid leukemia relatively late in life (from 12 to 15 months). In contrast to bone marrow cells from PLZF-RARalpha transgenic mice, those from NPM-RARalpha transgenic mice could be induced to differentiate by all-trans-retinoic acid (ATRA). We also studied RARE binding properties and interactions between nuclear corepressor SMRT and various fusion proteins in response to ATRA. Dissociation of SMRT from different receptors was observed at ATRA concentrations of 0.01 microM, 0.1 microM, and 1.0 microM for RARalpha-RXRalpha, NPM-RARalpha, and PML-RARalpha, respectively, but not observed for PLZF-RARalpha even in the presence of 10 microM ATRA. We also determined the expression of the tissue factor gene in transgenic mice, which was detected only in bone marrow cells of mice expressing the fusion genes. These data clearly establish the leukemogenic role of PLZF-RARalpha and NPM-RARalpha and the importance of fusion receptor/corepressor interactions in the pathogenesis as well as in determining different clinical phenotypes of APL.

[Cloning and identification of recombinant cDNA to a rabbit oviductin "DPF-1"].

A recombinant cDNA library to polyA + RNA isolated from rabbit oviduct epithelial cells was constructed, and screened with a polyclonal antibody against DPF-1 (64 kDa). 4 immunopositive plaques (DPF-1.1, DPF-1.2, DPF-1.3 and DPF-1.4) were purified. The polyclonal antibodies were epitope-selected respectively against the fused proteins produced by these positive recombinant plaques. Identification of recombinant clones by epitope selection revealed that the epitope-selected antibodies from DPF-1.1, DPF-1.2 and DPF-1.3 could recognise not only DPF-1, but 44 kDa protein also (Fig. 2). By using EcoRI-Not1 digestion method, the insert cDNA fragment size of these three recombinants was revealed to be 0.8 kb, 1.2 kb and 1.2 kb respectively (Fig. 3). These cDNA fragments were then isolated and subcloned into pBluescriptKS, and recombinant plasmids (pDPF-1.1, pDPF-1.2 and pDPF-1.3) were constructed (Fig. 4). Dot blot hybridization with a 32p-labeled 1.2 Kb-insert of cDNA from pDPF-1.3 indicated that these recombinant plasmids could cross-hybridized (Fig. 5), further indicating that they all possessed a common nucleic acid sequence. Dot and Northern blotting analysis of total RNA prepared from eight different tissues (skeleton muscle, heart, kidney, oviduct, liver, spleen, lung and small intestine) showed that the gene encoding DPF-1 was expressed specifically in the oviduct tissue (Fig. 6, Fig. 7).

[Preliminary studies on the rabbit oviductin "DPF-1"].

Anti-rabbit 64 kDa oviductin (named Development Promoting Factor-1, DPF-1) antibody could inhibit totally the early development of mouse fertilised eggs cultured in the conditioned medium derived from the rabbit oviduct mucosa epithelial cells, revealed that DPF-1 synthesized and secreted from rabbit oviduct mucosa has a function to overcome the developmental block of early mouse embryos. It seems that DPF-1 consists of a group of polypeptide isoforms, since its isoelectric points are ranging from 7.2 to 8.1 (Fig. 3). The synthesis and secretion of DPF-1 was not dependent on either 17 beta-estradiol or progesterone (Fig. 7), it can pass through zona pellucida easily and associate tightly with the early embryonic cell membrane (Fig. 6). By using Western blotting method, we found that DPF-1 was not appeared in the tissues of liver, heart, lung, spleen, uterus, ovary, small intestine, skeleton muscle and brain, but in that of oviduct (Fig. 4): some DPF-1 homologous molecules were also revealed in the oviduct tissues of mouse and golden hamster, their apparent molecular weights were 32 kDa, 72 kDa in mouse, and 49 kDa, 68 kDa in golden hamster (Fig. 5). Results obtained from the in vivo anti-fertility experiment, namely to analyse the anti-fertility effect in adult female mice after active immunization with DPF-1, showed that the fertility decreased significantly as compared to those of controls (p < 0.01) (Table 1). DPF-1 and its in vivo "loss of function" evidence we obtained will encourage us to study the mechanism of DPF-1 in overcoming the developmental block of early embryos, and its role in transition from maternal to embryonic control of early development.

[A preliminary study on early development of goat (Capra hircus) reconstituted embryos].

Goat reconstituted embryos (REs) have been produced by electrofusion-mediated nuclear transplantation method. Single cell derived from normal embryos or REs developed to 8-cell morula stage or the inner cell mass (ICM) of early blastocyst stage was used to fuse with enucleated mature egg (26-28 hrs after injection of LRH). According to the results summarized in table 1 and 2, we decided to adopt the method to embed REs in agarose and then transfer into goat oviduct lumen of host mother for 4-6 days in vivo culture. Normal fertilized eggs seem to develop synchronously, but that of REs are not. Tables 4 and 5 reveal that REs and embryos reconstituted successively can develop normally, no significant difference was found among their development rates. All these experimental results indicate that nuclei of some blastomeres from normal embryos or REs (derived from eight cells to morula or ICM) retained their totipotency for further development. These nuclei can be reprogrammed in host ooplasm, and developed to term.

[Emancipation of the developmental block of early rat embryos by protein factors excreted from rabbit oviduct epithelial cells in culture].

Experiments were designed to evaluate the ability of rabbit oviduct epithelial cells (ROEC) or ROEC conditioned medium to promote the development of rat eggs fertilized in Vivo or in Vitro. 61.73% of the eggs fertilized in Vitro and 73.33% of the eggs fertilized in Vivo cocultured with ROEC overcame the 2-cell block (Plate I, tables 1 and 2). Similarly, 67.99% of the eggs fertilized in Vitro cultured in ROEC conditioned medium developed over the 2-cell stage, and nearly half of them developed to morula and blastocysts stage (Table 3). By using 35S-methionine incorporation and autoradiography methods, several polypeptides, with molecular weight of 135 Kd, 68 Kd, 55 Kd, 51 Kd and 44 Kd respectively (Fig. 1), were excreted from rabbit oviduct epithelial cells and found in the ROEC conditioned medium. The possibility of entrance of the ROEC proteins into the developing embryo was tested by determining whether any of the secreted proteins bound to the zona pellucida. The results of iodination by using 125I-containing acylating agent labelling method showed that the 68 Kd and 55 Kd proteins were bound onto the zona pellucida of rat eggs co-cultured with ROEC in vitro for 24 h (Fig. 2). Studies concerning the problem that whether these two secreted proteins are the key factors to promote the development of early embryos and the transition of maternal to zygotic control of embryo development are undertaking.