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Objective: To study the effect of triple organ transplantation (liver, kidney, and pancreas) in patient of end-stage liver disease with renal failure and diabetes, and to explore the optimal surgical procedure., Methods: Simultaneous piggyback orthotopic heterotopic liver, pancreas-duodenum, and kidney transplantation was performed on a 43-year-old male patient with exocrine pancreatic insufficiency and insulin-dependent diabetes related to chronic pancreatitis (CP) who developed hepatic and renal failure. The pancreatic exocrine secretions were drained enterically to the jejunum. Prednisone, tacrolimus, mycophenolate mofetil, and ATG were used as immunosuppression therapy., Results: Good liver and pancreas allograft function recovery was achieved within 7 days after the operation. And the recovery of renal allograft function was delayed. The renal allograft was removed because of break-down of renal blood flow 16 days after the transplantation. A new renal transplantation was performed at the same position. The second kidney graft recovered its normal function 3 days later. Up to the writing of this paper no acute rejection of organs and such complications as pancreatitis, thrombosis, and localized infection occurred. The patient became insulin independent with normal liver and renal function., Conclusion: Simultaneous piggyback orthotopic heterotopic liver, pancreas-duodenum, and kidney transplantation can be a good method for the patients with exocrine pancreatic insufficiency and insulin-dependent diabetes combined with hepatic and renal failure.

Taking magnesium oxide slurry as absorption solution, the simulation of bubbling absorption process of mixed SO2 gases was observed in laboratory. Experiment results show that with a high efficiency and stable situation, acidification of absorbing solution was caused by HSO3-; the acidification trend was in accordance with the pattern of hydrolyzing of SO2, pH changes slowly at high pH value with SO3(2-) and rapidly at low value with HSO3-. The experiments also show the insensitive effect of liquid temperature on the high desulphurization efficiency. With relatively high dissolution rate and oxidizability of MgSO3 as well as the high solubility of MgSO4, the desulphurization efficiency utilization of MgO. Industrial experiment of FGD of coal-fired boiler showed that by recycling absorbing liquid could be raised to the concentration of MgSO4 to the saturation concentration at the operation temperature (40-50 degrees C) without any adverse effects on FGD efficiency. Refinement and enrichment of active substance could promote the desulphurization process, thus showed the availability of technical and economy feasibility of recovery technology.

The incomplete correlation between the organismal complexities and the number of genes among eukaryotic organisms can be partially explained by multiple protein products of a gene created by alternative splicing. One type of alternative splicing involves alternative selection of mutually exclusive exons and creates protein products with substitution of one segment of the amino acid sequence for another. To elucidate the evolution of the mutually exclusive 115-bp exons, designated flip and flop, of vertebrate AMPA receptor genes, the gene structures of chordate (tunicate, cephalochordate, and vertebrate) and protostome (Drosophila and Caenorhabditis elegans) AMPA receptor subunits were compared. Phylogenetic analysis supports that the vertebrate flip and flop exons evolved from a common sequence. Flip and flop exons exist in all vertebrate AMPA receptor genes but only one 115-bp exon is present in the genes of tunicates and cephalochordates, suggesting that the exon duplication event occurred at the ancestral vertebrate AMPA receptor gene after the separation of vertebrates from primitive chordates. The structures of animal AMPA receptor genes also suggest that an intron insertion to separate the primordial flip/flop exon from the M4-coding exon occurred before the exon duplication event and probably at the chordate lineage.

Objective: To investigate the effect of antisense oligonucleotide (ASODN) targeting survivin on the apoptosis and proliferation of renal cancer cell line 786-O and enhancement of its sensitivity to epirubicin., Methods: ASODN targeting survivin was designed and constructed. Cultured cells were divided into 6 groups: control group, liposome group, sense oligonucleotide (SODN) group, 600 nmol/L ASODN group, and 600 nmol/L ASODN combined with epirubicin group. After transfected for 24 h, cultured cells were harvested to carry on the next tests. Cell morphological changes were examined by transmission electron microscopy. Survivin protein was detected by immunohistochemical method. Apoptosis index (AI) and proliferation index (PI) were examined by flow cytometry., Results: Morphological abnormalities of cells were observed in ASODN transfected groups. Expression of survivin in ASODN groups were significantly decreased compared with that in the control group, liposomes group and SODN group. AI of ASODN groups was significantly higher than that in other groups. PI of ASODN groups was significantly lower than that in other groups. The PI of ASODN combined with epirubicin group was (35.7 +/- 1.67)%, but (9.3 +/- 0.34)% or (8.5 +/- 0.21)% in liposomes group or SODN group that had combined with epirubicin. The ASODN group achieved the strongest effects to enhance apoptosis in comparison with control group (P < 0.05), while SODN did not cause statistically significant change (P > 0.05)., Conclusion: The expression of survivin protein in the renal clear cell carcinoma cell line 786-O is downregulated by survivin ASODN. ASODN targeting survivin induces apoptosis and inhibits proliferation of 786-O cells. Inhibition of survivin enhances sensitivity of 786-O to epirubicin.

Objective: The purpose of this study was to evaluate the clinical result of 546 tetracycline-stained teeth restored with a porcelain laminate veneer system (Cerinate, Den-Mat, USA) for aesthetic reasons., Methods: Tetracycline-stained teeth (546) were restored with a porcelain veneer system, and bonded with Ultra Bond resin cement. The restorations were recalled after 0.5, 1.5 and 2.5 years, respectively. Modified Ryge criteria were used to evaluate the veneers marginal adaptation, interfacial staining, secondary caries, postoperative sensitivity and the patients' satisfaction of the colour of the restorations., Results: This study found that 99% veneers had excellent marginal adaptations; and less than 1% veneers required rebonding in the first 6 months; the colour of the veneers was stable and no evident staining was found. Almost all patients were satisfied with the colour match of their restorations 1 year after placement., Conclusions: The research indicated that the porcelain veneer restoration system under investigation provided a reliable and highly satisfactory choice for the aesthetic restoration of tetracycline-stained teeth.

To find out if it was possible to prefabricate bone graft in the shape of a human mandibular ramus that possessed a pedicle that carried blood. The pore size of the natural coral was about 200 microm with a porosity of about 36%. The natural coral was made into the shape of a human mandibular ramus. Marrow-derived osteoblasts were seeded into porous natural coral scaffolds in a density of 2 x 10(8)/mL in 300 microL cell suspension. After two days of in vitro incubation, five cell-coral complexes were implanted into cell donor rabbits under the inferior epigastric blood vessels to prefabricate a vascularised bone graft of specific shape. Two months later the bone formation was observed by gross inspection and histological examination. Two months after operation, a well-vascularised bone graft in the shape of the initial coral scaffold and with a blood-carrying pedicle had been regenerated successfully. Osteogenesis followed the pattern of endochondral bone formation. New bone could be seen on the surface and in the pores of coral on histological examination. We have shown that it is possible to fabricate vascularised bone graft in a predetermined shape using tissue engineering. This kind of bone graft may have future clinical application.

Objective: To study the diagnosis and management of renal angiomyolipoma (RAML), and to identify risk factors affecting spontaneous angiomyolipoma rupture., Methods: The data of 68 patients with RAML from 1989 to 2002 were retrospectively reviewed. These patients were divided in two groups on the basis of tumor size, 35 patients in group A ( 4 cm)., Results: Seven patients were identified by image-guided percutaneous biopsy, and no major complications was noted. Sixteen patients with RAML were examined with angiography and 9 of 16 patients had got spontaneous rupture. 41.2% of patients were symptomatic, 4 cases (11.4%) in group A and 24 (72.7%) cases in group B (P < 0.01). There were significant differences in mean tumor size (11.6 cm +/- 5.1 cm vs 5.3 cm +/- 2.9 cm, P < 0.01) and mean aneurysm size (13.6 mm +/- 5.8 mm vs 2.6 mm +/- 3.0 mm, P < 0.01) between 9 cases of the ruptured tumor and 59 cases of unruptured tumor, 9 cases of the ruptured and 7 cases of unruptured tumor with angiography, respectively. Treatment consisted of conservative observation in 10 patients (no radiographic changes during the follow-up of 2 - 7 years); partial nephrectomy in 14 patients, tumor enucleation in 30 patients, total nephrectomy in 14 and posterior laparoscopic nephrectomy in 3 (no recurrence and complication correlation to operation during the follow-up of 2 - 144 months)., Conclusions: It is an important role that percutaneous biopsy guided by ultrasonography or computerized tomography performs in managing suspicious and/or indeterminate RAML. A higher probability of rupture is related to tumor and/or aneurysms size. Nephron-sparing surgery is the first choice for surgical treatment of RAML.

Temperature-dependent polymerising polyethylene oxide hydrogel was used as a vehicle to deliver bone marrow mesenchymal cells by injection in six nude mice, four mice acting as controls, to study generation of new bone in the cell-hydrogel complex. Mesenchymal cells were harvested by in vitro cell culture, and cells were seeded into polyethylene oxide solution. The density of the suspension was adjusted to 5 x 10(7)ml(-1). The hydrogel was obtained by adjusting the temperature to over 6 degrees C. Aliquots of 0.5 ml of the cell-hydrogel complexes were injected subcutaneously into the backs of the six experimental mice, and 0.5 ml of hydrogel alone was injected into the four controls. Generation of new bone was studied by gross inspection, radiographs, and histological examination. Two months after injection hard nodes had formed subcutaneously in all six mice, whereas in the control group the hydrogel had been absorbed completely and only soft tissue was present at the site of injection. A shadow could be seen on the radiographs of all cell-seeded mice. On histological examination of the nodes there was trabecular bone and some areas of neocartilage. This method of generating new bone might be of potential clinical use.

The amino acid, either a glutamine (Q) or an arginine (R), at the Q/R site of the pore-lining segment (M2) of a vertebrate AMPA receptor subunit critically influences the properties of the receptor. The R codon of the mammalian AMPA receptor subunit 2 (GRIA2) transcript is not coded by the chromosomal sequence, but is created by posttranscriptional RNA editing activities. On the other hand, the R codons of some teleost GRIA2 homologs are coded by chromosomal sequences. To elucidate the evolution of the utilization of Q/R RNA editing in modifying vertebrate GRIA2 transcripts, the GRIA2 genes of five fish species and an amphibian were studied. The putative hagfish GRIA2 homolog (hfGRIA2) encodes an R codon, whereas shark and bullfrog GRIA2 genes specify a Q codon at the genomic Q/R site. All gnathostoma GRIA2 genes possess an intron splitting the coding regions of M2 and the third hydrophobic region (M3). The intronic components required for Q/R RNA editing are preserved in all the Q-coding vertebrate GRIA2 genes but are absent from the R-coding GRIA2 genes. Interestingly, the hfGRIA2 is intronless, suggesting that hfGRIA2 is unlikely evolved from a Q/R editing-competent gene. Results of this study suggest that modification of GRIA2 transcripts by Q/R editing is most likely acquired after the separation of the Agnatha and Gnathostome.

The AMPA receptor (AMPAR), a pharmacologically defined ionotropic glutamate receptor, mediates fast excitatory synaptic transmission in the vertebrate central nervous system. Mammalian and avian AMPARs are assembled from the products of four genes (GRIA1-GRIA4) conserved in their translated sequences and gene organizations. Teleost fish also express AMPAR subunits; however, the AMPAR genes have not been extensively investigated in lower vertebrates. To elucidate the evolution of vertebrate AMPAR genes, reverse-transcriptase PCR-based surveys of subunits expressed in the brains of eight nonmammalian vertebrates were performed. The newly cloned vertebrate AMPAR subunits were classified by their sequence identities to the mammalian AMPAR subunits. The results of molecular and phylogenetic analyses indicated that the members of the AMPAR gene family increased from two in the jawless hagfish to four in the tetrapods and the shark and to more than four in the teleost fish. The sizes of AMPAR gene families correlate well with those of many multigene families observed in various vertebrates. Moreover, all vertebrates expressed at least one AMPAR subunit bearing an arginine (R) at the Q/R site, at which no invertebrate glutamate receptor subunit has been found to have an R residue, suggesting that the low calcium-permeable AMPARs appeared at early evolutionary stages of vertebrate central nervous systems. Uniquely, the loop 1 (L1) regions between hydrophobic domain 1 and hydrophobic domain 2 of the hagfish putative GRIA2 and all the teleost GRIA1 subunits were much longer than those of the remaining known ionotropic glutamate receptor subunits. The length and sequence of the L1 of teleost GRIA1 subunits were heterogeneous, suggesting that the amino acid residues in L1 were not highly selected.

The aim of the present study was to determine the relationships between bone morphogenetic proteins (BMPs), BMP receptor type IA and carcinogenesis of oral epithelium. A retrospective study was performed on material obtained from oral mucosa, including nine cases of normal mucosa (NB), eight cases of nonspecific chronic inflammation (NCI), seven cases of hyperkeratosis (HK), five cases of squamous cell papilloma (SCP), 29 cases of squamous cell carcinoma (SCC) with various grades of differentiation and 10 cases of epithelium adjacent to carcinoma (EAC). Six cases of NB from hard palate (NHP) were chosen as a control group. The benign groups consisted of NCI, HK and SCP. The antibodies against BMP-2/4, -5, receptor BMPR-IA and purified bovine BMP (bBMP-McAb) were utilised using an immunocytochemical method. The results demonstrated that the immunostaining of BMP-2/4, BMP-5, BMPR-IA and bBMP-McAb was weak and not consistent in normal and benign groups. The immunoreactivity level was independent of the clinical and pathological grading of SCC. All cases of SCC showed positive staining for BMP-2/4, BMP-5, BMPR-IA and bBMP-McAb except for three cases and one case of SCC which negatively stained for BMP-2/4 and BMP-5, respectively. The staining intensity and proportion of the positively stained cells were markedly increased in SCC when compared with that of the normal and benign groups except for EAC. The metastatic carcinoma cells in lymph nodes were strongly and positively stained for BMP-2/4 and BMP-5 when compared with the primary lesions. Our results indicate that there was an overexpression of BMP-2/4, BMP-5, bBMP-McAb and BMPR-IA in the high-risk premalignant and malignant lesions of oral epithelium. Our findings suggest that BMP-2/4 and BMP-5 but not BMPR-IA might be involved in the metastasis of oral carcinoma cells.

Objective: To evaluate, in vitro, the cytotoxicity of the herpes simplex virus thymidine kinase (HSV1-tk) gene and ganciclovir (GCV) treatment, a widely used prodrug/suicide gene therapy, in human tongue cancer cell line Tbp-TL., Methods: Retrovirus expressing plasmid-liposome complex transduction was used. After treatment with G418 for 3 weeks, the integrated HSV1-tk gene was detected by PCR test with the primers specifically designed for HSV1-tk cDNA; HSV1-tk mRNA was detected with in situ hybridization method. In vitro cytotoxicity effect of HSV-GCV on Tbp-TL was detected with MTT and trypan blue exclusion test., Results: In vitro experiments demonstrated dose- and time-dependent cell death by transduction of the HSV1-tk gene followed by GCV treatment. The IC50 (the concentration required to elicit 50% growth inhibition) shifted from 1000 micrograms/mL in non-transduced Tbp-TL cells to 1.1 micrograms/mL in Tbp-TL/TK cells transduced with HSV1-tk gene, with the therapeutic index of 1000. Treatment with GCV at a dose of 10 micrograms/mL for 5 days led to complete cell death in HSV1-tk transduced tumor cells. When mixed with tk positive cells, the non-transduced cells (both the parent Tbp-TL cells as well as another non-transduced cell, the human muco-epidermoid carcinoma cell line M3SP2) were also killed by a degree associated with the percentage of tk positive cells. Seventy to 90% of the mixed cells were killed with only 10% to 20% tk positive cells involved., Conclusion: These results suggest that the HSV-tk/GCV approach to human tongue cancer cell line may be efficacious, with a wide therapeutic range, and that it exerts a bystander effect in vitro.

Objective: To test the effectiveness of the new techniques of tissue-engineered cartilage., Methods: Chondrocytes were harvested through type II collagenase digestion from the auricle of New Zealand rabbits. The cells were mixed with alginate to generate chondrocytes/alginate composites with final cellular density of 50 x 10(6) per mL. Calcium chloride was used as the cross-linking agent to gel the aqueous alginate solution. The chondrocytes/alginate composites were injected into the dorsal subcutaneous tissue of New Zealand rabbits through autologous cells grafts. The specimens were observed during cartilage formation at 4, 8, and 12 weeks after injection., Results: Prior to harvesting, chondrocytes/alginate composites were easily visualized under the dorsal skin of animals. The appearance of experimental specimens was similar to that of native cartilage in gross morphology. Using a standard hematoxylin and eosin stain, the histologic features of all experimental specimens demonstrated new cartilage formation. With a Masson's trichrome and safranin O stain, the presence of collagen and glycosaminoglycan (GAG) was observed at 8 and 12 weeks., Conclusion: This study demonstrated that polymerization of alginate hydrogel can be controlled to allow injection of chondrocytes that produce new autologous cartilage at subcutaneous dorsal site of rabbits. Injectable tissue-engineered autologous cartilage is promising for potential use in oral and maxillofacial surgery.

Objective: Fibronectin (FN) has been shown to play a crucial role in odontoblast differentiation, and it is involved in the repair of dentine and pulp. The responsiveness of human dental pulp (HDP) cells to bovine plasma FN was investigated., Materials and Methods: After isolation and selection noncarious freshly extracted third molars in vitro, HDP cells were incubated for 72 hours one of four concentrations (10, 20, 40, 80 micrograms/ml) of bovine plasma FN with Dulbecco's modified eagle medium (DMEM) containing 1% fetal calf serum (FCS). We examined cell proliferation (using an MTT assay), DNA synthesis (using incorporation of [3H]-thymidine), the cell area, and alkaline phosphatase (ALPase) activity of the culture supernatant. The expression of FN, types I and III collagens, binding of lectins concanavalin(ConA), or wheat germ agglutinin (WGA) in vitro was examined by means of immunocytochemistry. The expression degree and the cell area were analyzed semi-quantitatively with an image processing and analysis system., Results: After 72 hours, HDP cells appeared contracted and smaller in the absence of FN when compared to cells incubated with FN. HDP cells were in spread state with all four concentrations of FN. FN (20, 40 micrograms/ml) stimulated cell proliferation, promoted incorporation of [3H]-thymidine, and increased the levels of ALPase activity in supernatant. The binding of ConA, the expression of type I collagen, and FN were significantly increased, while the binding of WGA, and the expression of type III collagen were significantly decreased in the FN (40, 80 micrograms/ml) groups when compared to the FN (10, 20 micrograms/ml) groups., Conclusion: These findings suggest that HDP cells utilize FN for attachment and subsequent spreading. FN might have mitogenic activity and some role in the regulation of differentiation of pulp cells into odontoblasts.

Objective: Tooth enamel is formed by ameloblasts, which are derived from epitheliums and secrete an extracellular matrix containing a complex arrangement of protein components. The epithelial component, referred to as the enamel organ, contains a layer of cells that secrete an organic matrix that biomineralizes to become tooth enamel. Adjacent ectomesenchyme cells differentiate to become dentinproducing odontoblasts. These two mineralized matrices form the crown of the vertebrate tooth. Therefore, amelogenins play a critical role in tooth enamel formation. We have examined the expression patterns and tissue distribution of amelogenins in their developmental stages in order to build a foundation for further study., Methods: Amelogenin expression patterns and tissue distribution in developing teeth of embryonic (E17E19) and neonatal (1 to 9 days old) Wistar rats were examined by immunohistochemistry., Results: Positive immunostaining for amelogenin was first observed in the late embryonic stage, E18. The highest level of amelogenin was noted in neonatal secretary ameloblasts, fully engaged in enamel matrix deposition (3 to 5 days old). After that, amelogenin expression continued at a lower level (6, 7, 8 days old). There was no amelogenin staining observed in the maturation stage of development (9 days old)., Conclusions: Amelogenin expression occurs as early as the polarization stage of pre-ameloblasts. Amelogenin was also expressed, but at a low level, in post-secretary stages of amelogenesis. Odontoblasts did not contain detectable amelogenin.