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To investigate the distribution and density ofDuring the period from June to October in 2018 and 2019, six counties (districts, cities) were sampled in southern, northern and central Jiangsu Province as surveillance sites. The density ofA total of 104 423

To analyze the changes of small molecular metabolites in the larvae of a deltamethrin-sensitive strain ofThe 50% and 75% lethal concentrations (LCThe LCCarbohydrate, carboxylic acid and its derivatives, fatty acyls, amino acids and their derivatives may play important roles in deltamethrin metabolism in the larvae of a deltamethrin-sensitive strain of

To compare the complete mitochondrial genome sequences of two phenotypes ofFollowing infection with large and smallThe genetic distance between the phenotypes of

Recent experimental evidence points to the possibility that cell surface-associated caveolae may participate in mechanotransduction. The particular shape of caveolae suggests that these structures serve to prevent exposure of putative mechanosensors residing within these membrane invaginations to shear stresses at magnitudes associated with initiation of cell signaling. Accordingly, we numerically analyzed the fluid flow in and around caveolae using the equation of motion for flow of plasma at low Reynolds numbers and assuming no slip-condition on the membrane. The plasma velocity inside a typical caveola and the shear stress acting on its membrane are markedly reduced compared to the outside membrane. Computation of the diffusion field in the vicinity of a caveola under flow, however, revealed a rapid equilibration of agonist concentration in the fluid inside a caveola with the outside plasma. Western blots and immunocytochemistry support the role of caveolae as shear stress shelters for putative membrane-bound mechanoreceptors such as flk-1. Our results, therefore, suggest that caveolae serve to reduce the fluid shear stress acting on receptors in their interior, while allowing rapid diffusion of ligands into the interior. This mechanism may permit differential control of flow and ligand activation of flk-1 receptor in the presence of ligands.

The pathogenesis of tongue cancer (TA) has not been fully illustrated. Cyclooxygenase-2 (COX-2) is correlated with the precancerous lesion of oral cavity mucosa and malignant transformation. The focal adhesion kinase (FAK) and gap junction protein connexin 43 (Cx43) are involved in the occurrence and progression of tumors. This study aimed to investigate the effect of celecoxib on the proliferation, malignant transformation, and expression of FAK and Cx43 proteins.Healthy male Sprague-Dawley (SD) rats (4 months old) were divided into control, model and celecoxib group. 7,12-dimethylbenzanthracene (DMBA) was used to generate tongue mucosal carcinoma, coupled with celecoxib intervention. At 8, 12, 16, and 20 weeks after induction, the rat survival status, the tumor formation rate and the tongue tissue morphology were observed. Meanwhile, the expression of FAK and Cx43 was also evaluated by using immunohistochemistry (IHC).Tumor occurrence rates after induction were 0, 26.67%, 66.67%, and 80% at 8, 12, 16, and 20 weeks, respectively. The celecoxib treatment decreased such rats to 0, 0, 0, and 13.33%, respectively (p0.05 compared to model group). No significant change was observed in control group, whilst model group had mild to severe hyperplasia and squamous carcinoma with elongated time. Celecoxib treatment significantly improved the tissue morphology (p0.05). The model group also had elevated FAK and depressed Cx43 protein expression (p0.05). With elongated time, the FAK expression was further increased whilst Cx43 protein was depressed (p0.05 compared to model group).The focal application of celecoxib effectively inhibited the DMBA-induced rat TA, possibly via regulating FAK and CX43 protein expression, and inhibiting oral epidermal hyperplasia.

Colonic organoids are three-dimensional organotypic cultures of the colonic stem cells or pluripotent stem cells. Its essence is the culture of colonic stem cells or pluripotent stem cells, and their derived intestinal epithelial cells, intestinal endocrine cells and goblet cells in basement membrane extract with specific growth factors. Colonic organoids are comprised of all major types of colonic epithelial cells and represent the architecture and function remarkably similar to those of the colonic epithelium, faithfully recapitulating the functional colonic epithelium ex vivo. As a superior basic experimental model, colonic organoids are representing advantages over conventional cell models and animal models in many aspects, such as high successful rate, short productive cycle, and high consistency with source tissue. Since first reported in 2011, colonic organoids have soon become an important topic in the field of colonic diseases. It has now been applied in the field of physiology of colonic epithelium, infectious diarrhea, ulcerative colitis, regeneration of intestinal injury, and colon tumors. In this review, we summarize the research advances of establishment and application of colonic organoids.结肠类器官是一种衍生于结肠干细胞或多能干细胞的三维细胞培养模型，其实质是在添加多种生长因子的基质胶中对结肠干细胞或多能干细胞及其衍生的肠上皮细胞、肠内分泌细胞、杯状细胞等进行三维培养。体外构建的结肠类器官在细胞成分、组织架构及特定功能等方面与结肠上皮组织高度相似，实现了在体外培养环境中对结肠上皮组织的复制。与传统的结肠疾病研究模型（常规细胞培养模型和动物实验模型）相比，结肠类器官具有明显的优势，如培养成功率高、培养周期短、与来源组织高度一致等。自2011年Sato等首次报道结肠类器官模型的构建方法以来，结肠类器官迅速成为结肠相关疾病研究领域的热点。目前，结肠类器官已经被广泛应用于结肠上皮的生理功能的调整以及感染性腹泻、溃疡性结肠炎、肠道损伤再生、结肠肿瘤等多个领域的研究。本综述讨论了结肠类器官的构建及其应用研究的最新进展。.

The nasal-associated lymphoid tissues (NALTs), embedded in the submucosa of murine upper respiratory tract, represents an important site of induction for local mucosal immune responses to airborne pathogens and intranasal vaccines. Here, we systematically investigated the mucosal humoral and cellular immune responses of NALTs in mice infected with A/Beijing/501/2009 (BJ501) and A/Puerto Rico/8/1934 (PR8) viruses. Compared with PR8 infection, BJ501 induced a more rapid increase of virus-specific IgA and IgG antibodies in the nasal lavage fluid and a higher ratio of IgG1/IgG2a, indicating a stronger Th2 response to BJ501 in mucosal immunity. In addition, using virus-specific enzyme-linked immunospot assay (ELISpot assay), we observed higher and earlier responses of virus-specific IgA and IgG antibody-secreting cells (ASCs) and IFN-γ and IL-4 cytokine-secreting cells (CSCs) in NALTs of mice intranasally infected with BJ501 virus. In particular, the frequency of BJ501-specific IFN-γ-CSCs significantly correlated with the kinetics of BJ501 virus load in NALTs, suggesting an important role of IFN-γ-CSCs-associated mucosal cellular immune responses in BJ501 virus clearance. Collectively, BJ501 induced a more comprehensive and rapid mucosal immune responses in NALTs of mice, providing further understanding of the immune responses elicited by 2009 pandemic H1N1 virus in upper respiratory tract. Keywords: nasal-associated lymphoid tissues (NALTs); influenza virus; mucosal immune response; Th1/Th2 response.

Formaldehyde-based feed additives are approved in the US for Salmonella control and reducing bacterial contamination in animal feed. However, we hypothesize formaldehyde inclusion in swine diets may influence gut microbial composition due to its antimicrobial properties which might negatively influence microbial populations and pig growth performance. Also, formaldehyde inclusion in diets is known to reduce the dietary availability of amino acids. Therefore, our study was conducted to characterize if the effects of feed formaldehyde-treatment are due to influences on microbial population or diet amino acid (AA) sources. Dietary treatments were arranged in a (2 × 2) + 1 factorial with formaldehyde treatment (none vs. 1000 ppm formaldehyde) and crystalline AA inclusion (low vs. high) with deficient AA content plus a positive control diet to contain adequate AA content without dietary formaldehyde. Treating diets with formaldehyde reduced growth rate (P = 0.001) while the AA inclusion had no evidence of impact. Formaldehyde reduced feed bacterial content and altered fecal microbial communities (P

Glioblastoma multiforme (GBM) is the most common and aggressive brain tumour. The neoplasms are difficult to resect entirely because of their highly infiltration property and leading to the tumour edge is unclear. Gliadel wafer has been used as an intracerebral drug delivery system to eliminate the residual tumour. However, because of its local low concentration and short diffusion distance, patient survival improves non-significantly. Axl is an essential regulator in cancer metastasis and patient survival. In this study, we developed a controlled-release polyanhydride polymer loading a novel small molecule, n-butylidenephthalide (BP), which is not only increasing local drug concentration and extending its diffusion distance but also reducing tumour invasion, mediated by reducing Axl expression. First, we determined that BP inhibited the expression of Axl in a dose- and time-dependent manner and reduced the migratory and invasive capabilities of GBM cells. In addition, BP downregulated matrix metalloproteinase activity, which is involved in cancer cell invasion. Furthermore, we demonstrated that BP regulated Axl via the extracellular signal-regulated kinases pathway. Epithelial-to-mesenchymal transition (EMT) is related to epithelial cells in the invasive migratory mesenchymal cells that underlie cancer progression; we demonstrated that BP reduced the expression of EMT-related genes. Furthermore, we used the overexpression of Axl in GBM cells to prove that Axl is a crucial target in the inhibition of GBM EMT, migration and invasion. In an in vivo study, we demonstrated that BP inhibited tumour growth and suppressed Axl expression in a dose-dependent manner according to a subcutaneous tumour model. Most importantly, in an intracranial tumour model with BP wafer in situ treatment, we demonstrated that the BP wafer not only significantly increased the survival rate but also decreased Axl expression, and inhibited tumour invasion. These results contribute to the development of a BP wafer for a novel therapeutic strategy for treating GBM invasion and increasing survival in clinical subjects.

Clip domain serine proteases (CLIPs), characterized by one or more conserved clip domains, are essential components of extracellular signalling cascades in various biological processes, especially in innate immunity and the embryonic development of insects. Additionally, CLIPs may have additional non-immune functions in insect development. In the present study, the clip domain serine protease gene Bombyx mori serine protease 95 (BmSP95), which encodes a 527-residue protein, was cloned from the integument of B. mori. Bioinformatics analysis indicated that BmSP95 is a typical CLIP of the subfamily D and possesses a clip domain at the N terminus, a trypsin-like serine protease (tryp_spc) domain at the C terminus and a conserved proline-rich motif between these two domains. At the transcriptional level, BmSP95 is expressed in the integument during moulting and metamorphosis, and the expression pattern is consistent with the fluctuating 20-hydroxyecdysone (20E) titre in B. mori. At the translational level, BmSP95 protein is synthesized in the epidermal cells, secreted as a zymogen and activated in the moulting fluid. Immunofluorescence revealed that BmSP95 is distributed into the old endocuticle in the moulting stage. The expression of BmSP95 was upregulated by 20E. Moreover, expression of BmSP95 was downregulated by pathogen infection. RNA interference-mediated silencing of BmSP95 led to delayed moulting from pupa to moth. These results suggest that BmSP95 is involved in integument remodelling during moulting and metamorphosis.

Two experiments were conducted to investigate the effects of collection method and diet type on digestibility coefficients. In Exp. 1, 24 barrows were fed either a corn-soybean meal (CSBM) diet or CSBM with 20% dried distillers' grains with solubles (CSBM-DDGS). In Exp. 2, the effects of basal diet and collection method on determination of dried distillers' grains with solubles (DDGS) digestibility were studied using 24 barrows. The 4 diets used in Exp. 2 were: a CSBM (basal 1) , a barley-canola meal (BCM; basal 2), 80% basal 1 with 20% DDGS (CSBM-DDGS), and 80% basal 2 with 20% DDGS (BCM-DDGS). In both experiments, feces were collected using a time-based collection method (DY) or a "marker-to-marker" collection method (MM). Diets contained 0.5% of titanium dioxide (TiO) for estimating digestibility using the index marker approach (IM). The apparent total tract digestibility (ATTD) of DM and GE were lower (0.05) in the CSBM-DDGS diet than in the CSBM diet in Exp. 1 but were not different in Exp. 2. All the estimates of BCM-based diets were consistently lower (0.05) than those of CSBM-based diets. In Exp. 1, digestibility coefficients determined by the DY and MM were not different from each other, whereas those estimates were lower (0.05) using the IM than those using the total collection approach (TC; DY and MM). In Exp. 2, interactions (0.05) were observed between diet type and method for dietary digestibility coefficients. Digestibility and energy values estimated by the DY and MM were not different in pigs fed CSBM-based diets and the BCM-DDGS diet, whereas those estimates were greater (0.05) using the DY than those using the MM in pigs fed the BCM. There were no interactions between basal diet and method for estimating DDGS digestibility. The ATTD of DM and GE of DDGS using the MM were greater (0.05) than those using the IM, and ATTD of N tended to be greater (0.10) using the MM than that using the IM. All estimates using the DY were not different from those using the MM or the IM, except that DE of DDGS was greater (0.05) using the DY than when using the IM. Digestibility estimates of DDGS were not affected by basal diets. The mean DE and ME (as-fed basis) of DDGS were 3,994 and 3,688 kcal/kg, respectively, when estimated using the basal 1 diet and were 3,919 and 3,547 kcal/kg, respectively, when estimated using the basal 2 diet. In conclusion, both collection methods can be used to estimate energy and nutrient digestibility of diets and DDGS when using CSBM-based diets.

To observe the effects of San-huang-sheng-fu oil (S) on peripheral circulatory disorders and foot ulcers in diabetic rats and the relevant mechanisms.(1) Twenty-five Wistar rats were divided into non-diabetes (N), diabetes and sham treatment (DS), metformin (M), S, and combined treatment (CT) groups according to the random number table, with 5 rats in each group. Rats in group N were injected with sodium citrate buffer solution, while rats in the other 4 groups were injected with 10 mg/mL streptozotocin to induce diabetes. In post injection week (PIW) 3, feet of rats in all the 5 groups received an ice-cold stimulation to induce peripheral circulatory disorders. From PIW 9 to 12, rats in groups N and DS were gavaged with saline and applied with sesame oil on pelma of both hind limbs; rats in group M were gavaged with diluted M and applied with sesame oil on pelma of both hind limbs; rats in group S were gavaged with saline and applied with S on pelma of both hind limbs; rats in group CT were gavaged with diluted M and applied with S on pelma of both hind limbs. In PIW 9 before treatment (hereinafter referred to as before treatment) and post treatment week (PTW) 1, 2, and 3, plantar temperature and hot pain threshold of rats were detected by infrared thermometer and foot tester respectively. (2) Another 25 rats were divided and induced with diabetes (expect for group N) as above. In PIW 9, rats in the 5 groups were inflicted with foot ulcer in the left pelma of hind limb by steam and received the corresponding treatment. On post treatment day (PTD) 3, 7, 21, and 35, the general condition and area of wounds were observed and measured respectively. All the rats were sacrificed on PTD 35, and wound tissue was collected for histomorphological observation and determination of expressions of cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) using HE staining and immunohistochemical staining respectively. Data were processed with analysis of variance for repeated measurement, one-way analysis of variance, and Bonferroni post hoc test.(1) The experiment of peripheral circulatory disorders in diabetes. Compared with the plantar temperature of rats in group N, except for that in group CT in PTW 2 and groups M, S, and CT in PTW 3 (with t values from 0.258 to 2.647, P values above 0.05), the plantar temperature of rats with diabetes in the 4 groups at each time point was lowered significantly (with t values from 2.811 to 6.066, P values below 0.05). Compared with the plantar temperature of rats in group DS, except for that in group CT in PTW 2 and 3 significantly increased (with t values respectively 3.419 and 2.863, P values below 0.05), the plantar temperature of rats in groups M, S, and CT showed no significant difference at each time point (with t values from 0.128 to 1.654, P values above 0.05). The plantar hot pain threshold of rats was significantly decreased in group N than in the other 4 groups before treatment and group S in PTW 1 (with t values from 2.836 to 4.456, P values below 0.05). The plantar hot pain thresholds of rats in groups M, S, and CT were close to the hot pain threshold in group DS (with t values from 0.312 to 1.611, P values above 0.05). (2) The experiment of diabetic foot ulcers. Edema existed in all the wounds of rats on PTD 3. The wound areas of all the rats continued to increase with swelling and scar formation on PTD 7. On PTD 21, the scar of rats in groups N, S, and CT fell off; the wounds of rats in group DS were still swollen; scar of rats did not fall off with dark red in the skin around the wound in group M. On PTD 35, wounds of rats in groups N, S, and CT were nearly healed; while wounds of rats in groups DS and M were still swollen and the scar around the wound failed to fall off. On PTD 3 and 7, the wound areas of rats with diabetes in the 4 groups were close to those in group N (with t values from 0.111 to 1.476, P values above 0.05). On PTD 21, the wound area of rats in group DS was significantly larger than that in group N (t=5.502, P0.01), while the wound areas of rats with diabetes in the other 3 groups were close to the area in group N (with t values from 0.544 to 1.676, P values above 0.05). On PTD 21, the wound area of rats in group M was close to that in group DS (t=1.895, P0.05), while the wound areas of rats in groups S and CT were significantly smaller than the area in group DS (with t values respectively 5.809 and 3.426, P0.05 or P0.01). On PTD 35, the wound areas of rats in groups DS and M were significantly larger than the area in group N (with t values respectively 8.495 and 4.108, P values below 0.01), while the wound areas of rats in groups S and CT were close to the area in group N (with t values respectively 0.291 and 2.195, P values above 0.05). On PTD 35, the wound area of rats in group M was close to that in group DS (t=0.897, P0.05); while the wound areas of rats in groups S and CT were significantly smaller than the area in group DS (with t values respectively 6.923 and 6.583, P values below 0.01). On PTD 35, the structures of wound tissue were in better integrity with less inflammatory cells and more regularly arranged collagen fibers around the wounds of rats in groups N, S, and CT than in groups DS and M. On PTD 35, the expression levels of COX-2 and VEGF in the wounds of rats in group DS [respectively (222±89)% and (55±12)%] were close to those in group M [respectively (137±24)% and (94±36)%, with t values respectively 3.046 and 2.653, P values above 0.05]. On PTD 35, the expression level of COX-2 in the wounds of rats in group DS was significantly higher than the expression levels of COX-2 in groups N, S, and CT [respectively (100±35)%, (91±42)%, and (109±17)%, with t values from 4.039 to 4.653, P values below 0.01], while the expression level of VEGF in the wounds of rats in group DS was significantly lower than the expression levels of VEGF in groups N, S, and CT [respectively (100±28)%, (143±12)%, and (120±13)%, with t values from 3.363 to 5.905, P0.05 or P0.01].S can improve the plantar temperature decrease and pain dysesthesia of rats caused by diabetic peripheral circulatory disorders. It also can promote wound healing of diabetic foot ulcers in rats with down-regulation of COX-2 and up-regulation of VEGF.

Evaluating each patient and animal as its own control achieves personalized medicine, which honors the hippocratic philosophy, explaining that “it is far more important to know what person has the disease than what disease the person has.” Similarly, individualizing molecular signaling directly from the patient’s brain in real time is essential for providing prompt, patient-based treatment as dictated by the point of care. Fortunately, nanotechnology effectively treats many neurodegenerative diseases. In particular, the new medicinal frontier for the discovery of therapy for Parkinson’s disease is nanotechnology and nanobiotechnology. Indeed, the unique nanotechnology of neuromolecular imaging combined with the series of nanobiosensors enables continuous videotracking of molecular neurotransmitters in both the normal physiologic and disease states with long-term electrochemical operational stability. This nanobiotechnology is able to track a signal in real time with excellent temporal and spatial resolution directly from each patient’s brain to a computer as subjects are behaving during movement, normal and/or dysfunctional including prion-like Parkinson’s behavioral biometrics. Moreover, the molecular signaling performed by these nanobiosensors live streams directly online and originates from precise neuroanatomic brain sites such as, in this case, the dorsal striatum in basal ganglia. Thus, the nanobiotechnology studies discussed herein imaged neuromolecules with and without l-3,4-dihydroxyphenylalanine (l-DOPA) in dorsal striatal basal ganglia neurons. Parkinsonian and non-Parkinsonian animals were video-tracked, and images were readily seen on a laptop via a potentiostat using a semiderivative electrical circuit. Administered l-DOPA doses were 50 and 100 mg/kg intraperitoneally (ip); the same experimental paradigm was used to image and then contrast data. Results showed that the baseline release of biogenic amine molecules was significantly above detection limits in non-Parkinsonian animals. After administration of l-DOPA, biogenic amines significantly increased in these non-Parkinson’s animals. Nevertheless, it is intriguing to see that l-DOPA could not enable synaptic dopamine release in Parkinson’s animals, thereby demonstrating that biogenic amines are biomarkers for Parkinson’s disease. Biomarkers are biochemical, genetic, or molecular measures of biological reactions. Importantly, there were other significant biomarkers present in Parkinsonian animals and absent in non-Parkinsonian animals; these were peptide neurotransmitters that include dynorphin and somatostatin in the brain with detection limits of 40 nM for dynorphin and 37 nM for somatostatin (see Table 1). Furthermore, l-DOPA significantly increased these peptide biomarkers, dynorphin and somatostatin, in Parkinson’s animals. Targeting biomarkers enables new diagnostic devices and treatments for Parkinson’s disease through nanotechnology and nanobiotechnology.

A common feature of B-cell chronic lymphocytic leukemia (CLL) is chromosomal loss of 13q14, containing the miR15a/16-1 locus controlling B-cell proliferation. However, CLL etiology remains unclear. CLL is an adult leukemia with an incidence that increases with advancing age. A unique feature of CLL is biased B-cell antigen receptor (BCR) usage, autoreactivity with polyreactivity and CD5 expression, all suggest a role for the BCR in driving CLL pathogenesis. Among human CLLs, BCRs autoreactive with non-muscle myosin IIA (AMyIIA) are recurrent. Here we identify an unmutated AMyIIA BCR in mouse, with distinctive CDR3 segments capable of promoting leukemogenesis. B cells with this AMyIIA BCR are generated by BCR-dependent signaling during B-1 fetal/neonatal development with CD5 induction, but not in adults. These early-generated AMyIIA B-1 B cells self-renew, increase during aging and can progress to become monoclonal B-cell lymphocytosis, followed by aggressive CLL in aged mice, often with the loss of a chromosomal region containing the miR15a/16-1 locus of varying length, as in human CLL. Thus, the ability to generate this defined autoreactive BCR by B-1 B cells is a key predisposing step in mice, promoting progression to chronic leukemia.

Two experiments were conducted to evaluate the effects of spray-dried porcine plasma (SDPP) on growth performance, immunity, antioxidant capacity, and gut morphology of nursery pigs. In Exp. 1, 96 weaned pigs (Nebraska female × Danbred sire; 20 ± 1 d of age; initial BW = 6.06 ± 0.02 kg) were assigned to 16 pens and randomly allotted to the control (CTL; no SDPP) or the CTL + SDPP treatment in 2 phases (phase 1: d 0 to 14, 5% SDPP; phase 2: d 14 to 28, 2.5% SDPP). Blood samples were collected on d 0 and weekly thereafter to quantify IgG, IgA, and total antioxidant capacity. On d 14, pigs (n = 16; 8 pigs/treatment) were selected and euthanized for small intestine tissue and alveolar macrophage collection. On d 7, pigs fed SDPP had greater ADG, ADFI (P = 0.001), and G:F (P = 0.019) compared with CTL pigs. On d 28, pigs fed SDPP had greater BW (P = 0.024) and tended to have greater ADG (P = 0.074) and ADFI (P = 0.062) compared with CTL pigs. There were no differences between treatments for serum IgG, IgA, and total antioxidant capacity. On d 14, greater villus height (P = 0.011) and villus:crypt (P = 0.008) were observed in duodenal tissue sections obtained from SDPP-fed pigs compared with CTL pigs. To evaluate effects of SDPP on immune biomarkers, alveolar macrophages collected from 3 pigs/treatment on d 14 were cultured in vitro and challenged with lipopolysaccharide (LPS; 10 ng/mL). Therefore, 4 treatments included 1) CTL diet with no LPS, 2) CTL diet with LPS (CTL+), 3) SDPP diet with no LPS, and 4) SDPP diet with LPS. There were no diet effects on tumor necrosis factor-α gene expression or secretion by alveolar macrophages. For IL-10 gene expression, a diet × LPS interaction (P = 0.009) was observed where CTL+ had greater (P0.05) IL-10 mRNA abundance compared with other treatments. A second experiment was conducted to evaluate the in vitro effects of porcine plasma using model porcine jejunal epithelial cells (IPEC-J2). The treatments applied to the IPEC-J2 cells were 1) CTL, 2) CTL + 10% porcine plasma, 3) CTL + 5% porcine plasma, 4) CTL + 2.5% porcine plasma, or 5) CTL + 5% fetal bovine serum. The addition of porcine plasma increased (P0.001) proliferation of IPEC-J2 cells at 24 and 48 h following exposure to the respective treatments. In conclusion, dietary SDPP increased growth performance of nursery pigs during the first week postweaning and provide benefits to gut morphology possibly via increased proliferation of enterocytes.

Focal adhesions (FAs) are dynamic subcellular structures crucial for cell adhesion, migration and differentiation. It remains an enigma how enzymatic activities in these local complexes regulate their structural remodeling in live cells. Utilizing biosensors based on fluorescence resonance energy transfer (FRET), we developed a correlative FRET imaging microscopy (CFIM) approach to quantitatively analyze the subcellular coordination between the enzymatic Src activation and the structural FA disassembly. CFIM reveals that the Src kinase activity only within the microdomain of lipid rafts at the plasma membrane is coupled with FA dynamics. FA disassembly at cell periphery was linearly dependent on this raft-localized Src activity, although cells displayed heterogeneous levels of response to stimulation. Within lipid rafts, the time delay between Src activation and FA disassembly was 1.2 min in cells seeded on low fibronectin concentration ([FN]) and 4.3 min in cells on high [FN]. CFIM further showed that the level of Src-FA coupling, as well as the time delay, was regulated by cell-matrix interactions, as a tight enzyme-structure coupling occurred in FA populations mediated by integrin αvβ₃, but not in those by integrin α₅β₁. Therefore, different FA subpopulations have distinctive regulation mechanisms between their local kinase activity and structural FA dynamics.

Introduction We aimed to observe the trophic effects on graft structure and recipient metabolism of enteral rehabilitative therapy in rat small bowel transplantation (SBT). Methods Forty-eight recipients of rat allogeneic heterotopic SBT (Sprague-Dawley to Wistar rat) were divided into four groups randomly according to the presence of glutamine or growth hormone in the nutritional support regimen. We compared the histologic indices of graft mucosa and metabolic variables, including changes in body weight, nitrogen balance, urinary 3-methyl histidine excretion, and plasma albumin levels during 14 days after transplantation. Results Glutamine and growth hormone promoted recovery of graft structure and improved recipient protein metabolism, as evidenced by indices of the enteral rehabilitative therapy. Conclusions Glutamine and growth hormone potentiated their effects in enteral rehabilitative therapy, which produced potent trophic effects on graft structure and recipient metabolism in rat SBT.

In the medial nucleus of the trapezoid body (MNTB), each principal neuron receives one large axonal ending (a calyx of Held) and many small endings. In this same region, microelectrode recordings show unusual 'unit' waveforms which have two components separated by about 0.5 ms. We show that the first component (C1) of such a waveform corresponds to a spike from the calyx of Held and that the second component (C2) corresponds to a spike from the MNTB principal neuron. There are two kinds of evidence for these correspondences. First, electrical stimulation of calyciferous axons in the contralateral trapezoid body evokes C1 spikes with latencies of 0.1-0.3 ms. These latencies are too short for there to be an intervening synapse and are consistent with C1 being a presynaptic spike. Second, shocks in the lateral superior olive (which receives projections from MNTB principal-neurons) evoke 'A' spikes in the MNTB which can be shown by their waveshapes and mutual refractoriness with C2 spikes to result from antidromic activation of the neurons producing C2 spikes. Spontaneous and sound-evoked responses in dozens of cats anesthetized by barbiturates or Ketamine always had a C2 spike following each C1 spike. This implies that there is normally one-to-one spike transmission from the calyx of Held input to the MNTB principal neuron output. The small endings on MNTB principal neurons are also capable of evoking spikes. Electric shocks (and in one case, sound), evoked long latency (1-3 ms) 'LC2' spikes, which (by mutual refractoriness and waveshape) are from the same neural elements as C2 and 'A' spikes. Since LC2 spikes are not preceded by C1 spikes, LC2 spikes must be mediated by small axonal endings on MNTB principal neurons. We found some evidence of inhibition, possibly recurrent inhibition, in MNTB principal neurons. In a few neurons, sound or shocks inhibited 'A' spikes or LC2 spikes. In some cases, after each C2 spike, LC2 spikes were blocked or reduced in amplitude for several milliseconds. Our data firmly establish that there is fast, secure spike transmission from calyces of Held to MNTB principal neurons and suggest that under some circumstances there is additional signal processing in MNTB principal neurons.

Adhesions of cells to extracellular matrix and adjacent cells are mediated by integrins and VE-cadherin, respectively. Although these adhesion processes play crucial roles in vascular cell migration and angiogenesis, it remains unclear as to how they are coordinated to regulate cellular functions. We report here that integrin engagement by treating bovine endothelial aortic cell monolayers with beads coated with fibronectin (Fn) led to disruption of the VE-cadherin-containing adherens junctions. This disruption was accompanied by increases of tyrosine phosphorylation of β-catenin, γ-catenin, and p120 ctn , as well as the dissociation of α-catenin and γ-catenin from VE-cadherin. We applied a membrane-targeted Src reporter based on the fluorescence resonance energy transfer technique to visualize the dynamic Src activation at subcellular levels in live cells. The integrin engagement induced by Fn-coated beads caused the activation of Src around the beads and at adherens junctions, which are subsequently disrupted. The inhibition of Src with PP1 blocked the effects of integrin engagement on adherens junctions. Although Ras can also modulate adherens junctions, the resulting patterns of phosphorylation and association of junction proteins were distinct from those induced by integrin engagement. The inhibition of Ras by RasN17 did not rescue the disruption of adherens junctions induced by integrin engagement or by Src activation. Integrin engagement by Fn-coated beads also induced a significant alteration of cortical actin filaments at adherens junctions. The results indicate that integrin engagement disrupts VE-cadherin-containing adherens junctions via the activation of Src, but not Ras, possibly as a result of modulation of the actin network.

We have shown previously that anti-fecundity immunity can be induced experimentally against recombinant 26 kDa glutathione S-transferase (reSjc26GST) in Chinese water buffaloes (Bos buffelus), important reservoir hosts for Schistosoma japonicum in China. In the field study described here, we immunized buffaloes with reSjc26GST to induce protective immunity against S. japonicum and to evaluate its effectiveness in controlling schistosomiasis japonica. We selected two villages as test and control groups in inside-embankment areas endemic for schistosomiasis japonica. The buffaloes in the test village were vaccinated with reSjc26GST, whereas those in the control village were not. The indicators of the effect of the vaccine included the generation of specific IgG antibodies in the vaccinated buffaloes, changes in the prevalence and infection intensity in buffaloes and village children, changes in the density of infected snails, and changes in the infectivity of water bodies (assessed by sentinel mice) in transmission areas adjacent to both villages. Twenty months after vaccination, the infection rate of buffaloes in the test village was decreased by 60.4% (from an initial prevalence of 13.5% to 5.4%), and 67.9% when compared with that in the control village (initial prevalence of 16.7%). However, the infection rate in village children remained unchanged. The density of infected snails decreased by 71.4%, from 0.0049/0.11 m2 to 0.0014/0.11m2 in the high transmission area outside the embankment in the test village. There was no change in the infectivity of the water body transmission areas between the test and control villages. The levels of specific antibodies to reSjc26GST showed a continuous increase after vaccination. These results indicate that protective immunity was induced and maintained in buffaloes after vaccination with reSjc26GST. The vaccine could thus play a significant role in reducing S. japonicum transmission caused by water buffaloes in the Lake region of China.

Considering the narrow therapeutic index of digoxin and the low range between the safe and toxic serum concentrations of this drug, to evaluate the relative bioavailability of tablets and oral solution is necessary. The pharmacokinetic properties of digoxin after oral administration of its hydroxypropyl-beta-cyclodextrin (HPCD) inclusion complex to rabbits and human volunteers were investigated in comparison with those of commercially available tablets. The aqueous solubility of digoxin was enhanced by HPCD for about 2000 times at HPCD concentration of 50% (w/v). But in a human bioavailability study no significant difference was observed in the extent of absorption (AUC(0-t)) and Cmax between the two formulations. Time to reach peak was significantly shorter for the solution than for the tablets (p0.01). The pharmacokinetic results from the rabbit study were similar to human studies and no significant difference was observed for AUC, Cmax and Tmax. As the bioavailability of both tablets and solution is equivalent HPCD based oral digoxin solution could serve as an alternative to tablets.

To prove that Fujian Province is also a natural focus of Pagumogonimus veocularis(Pv).The adult worms were obtained from a cat fed with Pv metacercariae.Pv were found in Jianou, Fujian Province. All 1,873 Semisulcospira libertina showed negative. The positive rate of Tricula fujianensis and Erhaia jianouenesis were 0.10% (1/695) and 0.25% (5/2,038), respectively. The main crab host was S. fujianensis. Ps alone and mixed infection with Pv were found in the Sinopotamon, the infection rates were 36.8% (43/117) and 20.5% (24/117), respectively. The numbers of the metacercariae were 806 and 40, respectively. A cat was infected with 12 metacercriae of Pv, eggs were found in the stool 56 days after infection, and 6 worms were found in the lungs 68 days after infection.Fujian is one of the natural focus of Pv, cat is the adequate host. The fluke was identified as Pv according to the characteristics of the metacercariae.

In order to understand the determinants of schistosome-related hepato- and spleno-megaly better, 14,002 subjects aged 3-60 years (59% male; mean age = 32 years) were randomly selected from 43 villages, all in Hunan province, China, where schistosomiasis caused by Schistosoma japonicum is endemic. The abdomen of each subject was examined along the mid-sternal (MSL) and mid-clavicular lines, for evidence of current hepato- and/or spleno-megaly, and a questionnaire was used to collect information on the medical history of each individual. Current infections with S. japonicum were detected by stool examination. Almost all (99.8%) of the subjects were ethnically Han by descent and most (77%) were engaged in farming. Although schistosomiasis appeared common (42% of the subjects claiming to have had the disease), only 45% of the subjects said they had received anti-schistosomiasis drugs. Overall, 1982 (14%) of the subjects had S. japonicum infections (as revealed by miracidium-hatching tests and/or Kato-Katz smears) when examined and 22% had palpable hepatomegaly (i.e. enlargement of at least 3 cm along the MSL), although only 2.5% had any form of detectable splenomegaly (i.e. a Hackett's grade of at least 1). Multiple logistic regression revealed that male subjects, fishermen, farmers, subjects agedor = 25 years, subjects with a history of schistosomiasis, and subjects who had had bloody stools in the previous 2 weeks were all at relatively high risk of hepato- and/or spleno-megaly. In areas moderately endemic for Schistosoma japonicum, occupational exposure and disease history appear to be good predictors of current disease status among older residents. These results reconfirm those reported earlier in the same region.

To investigate the kinetics and the magnitude of intragraft gene expression of interleukin-2 (IL-2), interferon-gamma (IFN-gamma), perforin and granzyme B, and intragraft expression of interleukin-2 receptor (IL-2R) and intercellular adhesion molecule-1 (ICAM-1) during acute rejection episodes, and to analyze the changes in apoptosis in small intestinal allograft rejection.Heterotopic small intestine transplantation was performed with inbred rats F344/N (RT1l) and Wistar/A (RT1-Ak, RT1-Ed). All recipients were divided into four groups: group 1: Wistar, native control; group 2: Wistar--Wistar; group 3: F344--Wistar and group 4: F344--Wistar + cyclosporine A (6 mg.kg-1.d-1 i.m.). The grafts were harvested on postoperative days (PODs) 3, 5 and 7. All samples were examined pathologically. Intragraft mRNA expression of IL-2, IFN-gamma, perforin and granzyme B were detected with reverse transcriptase polymerase chain reaction (RT-PCR) and intragraft expression of IL-2R and ICAM-1 were stained using immunohistochemistry. We also analyzed the change in apoptosis rejection with terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL).Mild acute rejection occurred on POD 3 in the allograft group, moderate acute rejection on POD 5, and severe acute rejection on POD 7, while none of the isografts had histological evidence of acute rejection. Cyclosporine A could effectively control rejection. Gene expression was virtually negative in the native control. Only on POD 5 was IL-2 mRNA expression of allografts significantly higher than that of isografts (P0.05). IFN-gamma mRNA expression was significantly higher than that of the control groups (P0.01) on PODs 3, 5 and 7, and the level of perforin and granzyme B mRNA expression reached significantly higher levels than in the other two control groups on POD 5 and POD 7. Intragraft IL-2R expression of the allograft was significantly higher than that of the other three control groups. Only on POD 3 was intragraft ICAM-1 expression of allografts significantly higher than isografts. The number of apoptotic cells per crypt of allografts was significantly higher than that of the other three control groups on POD 3 and POD 5 (P0.01).Transcription of IL-2, IFN-gamma, perforin and granzyme B, and expression of IL-2R and ICAM-1 as well as apoptosis of epithelial cells of the grafts play an important role in small intestine allograft rejection. Intragraft gene expression of IFN-gamma and intragraft expression of IL-2R as well as apoptotic epithelial cells may become a specific and sensitive diagnostic method of clinical value. Furthermore, therapeutic strategies to alter these molecules in small intestine transplantation may improve the outcome of current antirejection therapy.

The migration of vascular endothelial cells (ECs) plays an important role in vascular remodeling. Here we studied the effects of cell morphology on the migration of bovine aortic ECs by culturing cells on micropatterned strips of collagen matrix (60-, 30-, and 15-microm wide). The spreading areas of the cells on 15- and 30-microm wide strips were 30% lower than those on 60-microm wide strips and unpatterned collagen. The cells on 15-microm wide strips completely aligned in the direction of the strip, and had significantly lower shape index than those in all other groups. On strips of all widths, ECs tended to migrate in the direction of strips. ECs on 15-microm wide strips had highest speed, particularly in the direction of the strip. Vinculin staining showed that the leading edge of ECs on 15-microm wide strips had focal adhesions that were oriented with their lamellipodial protrusion and the direction of cell migration; this arrangement of the focal adhesions may promote EC migration. The present study provides direct evidence on the role of cell morphology in EC migration, and will help us to understand the mechanisms of EC migration during angiogenesis and wound healing.

The expression of different sets of immunoglobulin specificities by fetal and adult B lymphocytes is a longstanding puzzle in immunology. In the past few years it has become clear that production of mu heavy chain and subsequent assembly with surrogate light chain to form the pre-B cell receptor complex is critical to promote development of adult B cell precursors in mouse bone marrow. Recently we found that instead of promoting pre-B cell expansion as in adult bone marrow, this complex inhibits pre-B cell growth in fetal liver, providing a previously unrecognized mechanism for alteration of the B cell repertoire with age. The consequence is very distinct primary repertoires for development of fetal B1 cells and adult bone marrow B2 cells.

Shear stress, the tangential component of hemodynamic forces, activates many signal transduction pathways in vascular endothelial cells. The conversion of mechanical stimulation into chemical signals is still unclear. We report here that shear stress (12 dynes/cm2) induced a rapid and transient tyrosine phosphorylation of Flk-1 and its concomitant association with the adaptor protein Shc; these are accompanied by a concurrent clustering of Flk-1, as demonstrated by confocal microscopy. Our results also show that shear stress induced an association of alphavbeta3 and beta1 integrins with Shc, and an attendant association of Shc with Grb2. These associations are sustained, in contrast to the transient Flk-1. Shc association in response to shear stress and the transient association between alphavbeta3 integrin and Shc caused by cell attachment to substratum. Shc-SH2, an expression plasmid encoding the SH2 domain of Shc, attenuated shear stress activation of extracellular signal-regulated kinases and c-Jun N-terminal kinases, and the gene transcription mediated by the activator protein-1/12-O-tetradecanoylphorbol-13-acetate-responsive element complex. Our results indicate that receptor tyrosine kinases and integrins can serve as mechanosensors to transduce mechanical stimuli into chemical signals via their association with Shc.

Photofrin photodynamic therapy (PDT) has recently received FDA approval for the palliative treatment of totally and partially obstructing esophageal malignancies. However, there is a need for new PDT photosensitizers because Photofrin has a number of undesirable features. The purpose of this study was to evaluate the efficacy of four amine-bearing silicon phthalocyanines--Pc4, Pc10, Pc12 and Pc18--as potential PDT photosensitizers. Equimolar concentrations of these Pc were found to be highly effective at causing the regression of RIF-1 tumors transplanted to C3H/HeN mice. The amount of Pc4 necessary to cause an equivalent amount of tumor regression in this model system was substantially less than the amount of Photofrin. The cutaneous phototoxicity of the silicon Pc photosensitizer was assessed by the utilization of the murine ear-swelling model. When C3H mice were exposed to 167 J/cm2 of polychromatic visible light from a UVB-filtered solar simulator, which emitted UV radiation and visible light above 320 nm, the Pc produced little, if any, cutaneous photosensitivity. These results indicate that Pc4, Pc10, Pc12 and Pc18 are at least as effective as Photofrin in PDT protocols, while at the same time addressing many of the drawbacks of Photofrin.

Sensitization to donor antigens is a problem of growing magnitude in clinical transplantation. At a molecular level, this is due to the interaction between antigen bearing antigen-presenting cells and recipient T cells and involves both antigen presentation and co-stimulation.Allogeneic islet transplantation was performed using DBA/2J donors and B6AF1 recipients. Four weeks before transplantation, recipient animals were given donor-specific transfusion (DST) alone, DST + CTLA4Ig, DST + control IgG, or no treatment. Graft loss was defined as a blood glucose300 mg/100 ml.Administration of DST + control IgG 4 weeks before transplantation resulted in accelerated rejection due to presensitization (median survival time of 8 days, compared with 14.5 days for the no-treatment group). Animals treated with CTLA4Ig in combination with DST had a median survival time of 12 days, compared with 8 days for DST + IgG.CTLA4Ig attenuates the tempo of accelerated rejection in this islet allograft model of presensitization, but does not prolong allograft survival as compared with no treatment.

Trophic factors are being increasingly recognized as important contributors to growth, differentiation, and maintenance of viability within the mammalian nervous system during development. Pleiotrophin (PTN) is a secreted 18-kDa heparin binding protein that stimulates mitogenesis and angiogenesis and neurite and glial process outgrowth guidance activities in vitro. We localized the sites and time course of expression of the Ptn gene and its protein product in developing and adult mouse nervous system. Expression of the Ptn gene was first observed at embryo day 8.5 (E8.5). At E12.5, transcripts of the Ptn gene were localized in developing neuroepithelium at sites of active cell division in the spinal cord and brain. At E15.5, transcripts were found in the somata of some but not all neurons and glia whereas in the adult its pattern of expression was nearly exclusively restricted to the brain. The PTN protein was found almost entirely in association with the axonal tracts during development and in adults. Furthermore, as opposed to the finding of PTN in both central and peripheral nervous systems during development, PTN was not expressed beyond the exit where axonal tracts become the peripheral nervous system in adults. At all sites and times examined, the somata that contained Ptn transcripts corresponded with the axonal tracts that contained the PTN protein. The results establish that Ptn is expressed in early development at sites of active mitogenesis in developing neuroepithelium and later in both glial cells and neurons at sites of neuronal and glial process formation in developing axonal tracts. The findings establish a correspondence in the localization of PTN within the nervous system at sites of normal developmental processes that correlate with the functional activities of PTN previously described in vitro.

Hemodynamic forces play a key role in inducing atherosclerosis-implicated gene expression in vascular endothelial cells. To elucidate the signal transduction pathway leading to such gene expression, we studied the effects of fluid shearing on the activities of upstream signaling molecules. Fluid shearing (shear stress, 12 dynes/cm2 [1 dyne = 10(-5)N]) induced a transient and rapid activation of p21ras and preferentially activated c-Jun NH2 terminal kinases (JNK1 and JNK2) over extracellular signal-regulated kinases (ERK-1 and ERK-2). Cotransfection of RasN17, a dominant negative mutant of Ha-Ras, attenuated the shear-activated JNK and luciferase reporters driven by 12-O-tetradecanoylphorbol-13-acetate-responsive elements. JNK(K-R) and MEKK(K-M), the respective catalytically inactive mutants of JNK1 and MEKK, also partially inhibited the shear-induced luciferase reporters. In contrast, Raf301, ERK(K71R), and ERK(K52R), the dominant negative mutants of Raf-1, ERK-1, and ERK-2, respectively, had little effect on the activities of these reporters. The activation of JNK was also correlated with increased c-Jun transcriptional activity, which was attenuated by a negative mutant of Son of sevenless. Thus, mechanical stimulation exerted by fluid shearing activates primarily the Ras-MEKK-JNK pathway in inducing endothelial gene expression.

The retinoid acid inducible midkine gene encodes a highly basic, heparin-binding protein that possesses potential functions of growth or differentiation during the early stages of mouse embryonic development. Midkine gene has been mapped to mouse Chromosome 2. Here we report that the midkine gene is located in close proximity to the m4 muscarinic acid receptor gene. M4 codes for a transmembrane G protein that functions to transduce the action of acetylcholine neural transmitters in the nervous system. The two genes have the directions of their transcriptions running toward one another, and the distance between their respective 3' ends is approximately 1.3 kb.

The Ames Salmonella/microsomal assay was employed to test the mutagenicity of some benzamines (aniline, and o- and p-phenylenediamine) and their nitro-derivatives (p-nitroaniline, 2-nitro-p-phenylenediamine, 3- and 4-nitro-o-phenylenediamine), using strains TA98 and TA100 and their nitroreductase-deficient mutants, TA98NR and TA100NR, in the presence and absence of rat S9 mix. The addition of the nitro-group to benzamine molecules converted them into direct mutagens. Furthermore, the position of the nitro-group affected their mutagenic activities. Cytotoxicity testing with Chinese hamster ovary cells (CHO-K1) showed that the presence of the nitro-group in these compounds had no specific effect on toxicity. The test compounds all showed a dose-related increase in inducing chromosomal aberrations in CHO cells. However, the presence of the nitro-group did not affect potency in inducing chromosomal aberrations. Compounds containing the nitro-group had higher initial oxidation potentials and dipole moments (mu) than their nonnitro-containing counterparts. The mutagenicity and toxicity of these compounds were not related to physico-chemical properties, including oxidation potential, energy difference (deltaE) between the lowest unoccupied molecular orbital (LUMO) and the highest occupied molecular orbital (HOMO), ionization potential (I.P.), and mu.

To study effects of 3-morpholinosydnonimine-N -ethylcarbamide (SIN-1) on hypoxia-induced mechanical and electric activities of the isolated pig coronary artery.Mechanical tension and membrane potential were measured simultaneously.Hypoxia initially caused a transient vascular smooth muscle cell membrane hyperpolarization followed by a membrane depolarization in isolated pig coronary artery. Subsequent addition of SIN-1 100 mumol . L-1 or verapamil (Ver) 10 mumol . L-1 led to membrane repolarization and relaxation of the vascular smooth muscle. Nitro-L-arginine (NLA) 0.2 mmol . L-1 and KCI 40 mmol . L-1 also induced membrane depolarization and vasoconstriction, which were similarly suppressed by SIN-1 or Ver.Hypoxic contractile response in isolated pig coronary artery is mediated by an increased Ca2+ influx via suppression of nitric oxide release.

The isolation and cloning of the ligand for the cytokine receptor, Mpl, have been recently described. In this report we present details of the purification of this novel cytokine (megakaryocyte growth and development factor [MGDF]) from aplastic canine plasma. Two forms of canine MGDF, with apparent molecular weights of 25 kD and 31 kD and sharing a common N-terminal amino acid sequence, were isolated. The sole contaminant detected in purified 25-kD or 31-kD MGDF was canine Ig. Canine MGDF is characterized as a human megakaryocyte colony-stimulating factor that acts synergistically with human recombinant stem cell factor but not interleukin-3. MGDF also appears to be physiologically regulated in response to platelet demand. In canine and murine models, serum levels of MGDF activity peak during the thrombocytopenic periods after irradiation, 5-fluorouracil, or antiplatelet antisera injections. These data indicate that the megakaryocyte-stimulating activity that accumulates in plasma in response to platelet losses is a novel cytokine that functions through an interaction with the Mpl cytokine receptor.

Expression of Ig transgenes in recombination-deficient mutant scid and Rag-1-mice results in the generation of pre-B and B cells, which are normally absent from these animals. In screening for protein tyrosine kinases (PTKs) that may play a role in this progression beyond the pro-B stage, we have identified five differentially regulated PTKs and compared their gene expression in defined stages of early B-lineage cells from normal, mutant, and Ig-transgenic mutant mice. Three PTKs (fgr, flk2/flt3, and tsk) show a comparable decrease at an early stage in all mice. In contrast, the decreasing expression of ret and the increasing expression of blk seen in differentiating B cells from normal mice are not observed in the mutant mice, unless they carry Ig transgenes. Therefore, our results show that the expression of certain PTKs is dependent on productive Ig rearrangement and suggest important roles for both Ret and Blk at distinct stages in the Ig-dependent progression of B cell differentiation.

Methylene chloride extracts of particulates from liquefied petroleum gas (LPG) combustion appliance were studied by using Ames test, micronucleus test and inducibility of pulmonary and hepatic aryl hydrocarbon hydroxylase (AHH) and glutathione S-transferase (GST) in rats. The extracts showed mutagenicity for Salmonella typhimurium strain TA98 and its derivatives TA98NR and TA98/1,8-DNP6 with or without S9 mix. The revertants in strains TA98NR and TA98/1,8-DNP6 were less than 40% and 50% of that in strain TA98 without S9 mix, respectively. Positive results were obtained in mouse bone marrow micronucleus assay. Intratracheal instillation of the extracts led to increase in pulmonary (but not hepatic) AHH and GST activities in rats. It was seen that AHH was more sensitive than GST to induction by the extracts.

Baroreflex sensitivity (BRS) in anesthetized rats was measured as the slope of the regression line of the concomitant maximal change in heart rate (delta R-R) and blood pressure (delta BP) induced by bolus i.v. phenylephrine or sodium nitroprusside, m-nisoldipine (m-Nis) i.v. 5, 10, 20 micrograms.kg-1 depressed the BRS (2.6 +/- 0.2, 2.4 +/- 0.3, 1.6 +/- 0.2, or 1.7 +/- 0.1, 1.6 +/- 0.2, and 1.4 +/- 0.2 ms.kPa-1, respectively) compared with before medication (P0.01). m-Nis may act directly on carotid sinus to depress the BRS, but not through cardiac beta or M receptor nor through central nervous system.

m-Nisoldipine caused a concentration-dependent depression of the contractile response and 45Ca influx evoked by KCl in isolated rat thoracic aorta. The IC50 value for contraction and 45Ca influx were 0.69 (95% confidence limits 0.32-1.5) nmol.L-1 and 0.35 (95% confidence limits 0.06-2.0) nmol.L-1, respectively. There was a positive correlation between the inhibition of KCl-evoked contraction and 45Ca influx (r = 0.996). m-Nisoldipine (0.1-10 mumol.L-1) did not influence the 45Ca influx into resting cells and failed to inhibit noradrenaline (1.0 mumol.L-1)-evoked contraction and 45Ca influx. The results suggest that the relaxant effect of m-nisoldipine on rat aorta may be closely related to the blockade of Ca2+ entry through a potential-dependent calcium channel.

Hydrochlorothiazide (HCT) (0.1, 0.3 mmol.L-1) inhibited the contraction of rat aortic strips induced by low (40 mmol.L-1), not higher concentrations of KCl. HCT (0.3 mmol.L-1) did not inhibit the CaCl2-induced contraction of the aortic strips depolarized with high K+ (KCl 80 mmol.L-1). The inhibitory effect of HCT (0.1 mmol.L-1) on KCl (20 mmol.L-1)-induced contraction was markedly antagonized by BaCl2 (0.1 mmol.L-1) and tetraethylammonium (TEA) (0.3 mmol.L-1), but not by glibenclamide (Gli, 0.01 mmol.L-1). With norepinephrine (NE) or 5-HT as agonists, HCT (0.3 mmol.L-1) also inhibited the contractions of rat aortic strips. In the 2 components of NE-induced contraction, HCT inhibited only the tonic component depending on Ca2+ influx, but not the phasic component elicited by the release of intracellular Ca2+. The inhibitory action of HCT was endothelium-independent. That the HCT (3 mmol.L-1) increased the 86Rb efflux rate coefficient was antagonized by BaCl2 (0.1 mmol.L-1), but not by Gli (0.01 mmol.L-1). The results indicated that the inhibitory effect of HCT on the contraction of rat aorta was attributable to the opening of membrane potassium channels.

Cerebral hypoxia-ischemia was produced in 7 day old rats by right common carotid artery occlusion combined with systemic hypoxia. In the hypoxic-ischemic group animals, striatal ACh contents on both sides were reduced to a significant extent in commensurate to AChE histochemical image analysis. All these changes could be restored to normal control by a protection dose (50 micrograms) of NGF intraventricularly injected.

The two step small-bowel transplantation was performed experimentally in pigs. Both ends of heterotopic segmental allografts of 100 cm small bowel were enterostomized during the first operation. After 4-6 weeks, the allograft was interposed in the continuity of the intestine. Rejection was developed in Group I (n = 6) without immunosuppression. The mean survival time of the grafts was 17.7 +/- 7.6 days, and recipients were alive after resection of necrotic grafts. The animals of Group II (n = 6) were treated by cyclosporine concomitantly with azathioprine and methylprednisolone. Two pigs were killed for severe pneumonia on day 92, 97 and the grafts were alive, the other four recipients and their grafts survived for more than 300 days, 270 days, 260 days and 260 days respectively. No GVHD was observed and rejection was the major problem in segmental intestinal transplantation.

The protein (PTN) encoded by the pleiotrophin (PTN) gene belongs to a recently described family of heparin-binding cytokines whose expression is temporally and spatially regulated during development. We have now isolated genomic clones of the human PTN gene, characterized its promoter region, determined its transcription initiation site(s), and established functional activity of the PTN promoter. A fragment -550/+191 that contains a CAAT box, no apparent TATA box, and four consensus sites for the binding of MyoD is sufficient to provide optimal promoter activity. A serum response element is found at -559 to -568. We also have identified the human PTN gene on chromosome 7, band q33 and the mouse Ptn gene on chromosome 6, respectively. The data thus identify and characterize the 5' end of the PTN gene and its promoter region, suggest potential regions that may contribute to the regulation of its transcriptional activity, and localize the PTN gene in human and mouse chromosomes.

Transgenic mice carrying the phosphoenolpyruvate carboxykinase promoter region-human growth hormone (PEPCK-hGH) fusion gene are characterized by accelerated growth and plasma hGH levels ranging from 100 to 700 ng/ml. Both sexes are fertile, in contrast to previous findings in metallothionein-I/hGH transgenic mice in which females are sterile, apparently due to luteal failure. Virgin transgenic PEPCK/hGH females from this line produce milk and can successfully raise foster litters to weaning. We conclude that the life-long presence of very large amounts of hGH in the circulation is compatible with ovulation, can override the effects of hGH-induced suppression of endogenous PRL release, and can support full lactation in animals that have not been primed by hormonal changes associated with pregnancy.

m-Nisoldipine (m-Nis) and nisoldipine (Nis) inhibited spontaneous contraction of isolated non-pregnant human myometrium, IC50 being 3.7 +/- 0.7 mumol.L-1 for m-Nis and 6.0 +/- 0.9 mumol.L-1 for Nis. Both drugs also suppressed spontaneous and oxytocin-induced contraction of isolated pregnant rat uterus. In isolated uterus from virgin rat, both drugs inhibited the contractions induced by KCl, acetylcholine (ACh), and oxytocin (Oxy), the values of pD'2 against KCl, ACh, Oxy being 9.0 +/- 0.2, 6.1 +/- 0.1, and 5.9 +/- 0.1 for m-Nis and 9.1 +/- 0.3, 5.7 +/- 0.2, and 5.6 +/- 0.1 for Nis respectively. The results indicate that both m-Nis and Nis may be clinically useful for the treatment of dysmenorrhea and premature labour.

The antihypertensive effects of m-nisoldipine (m-Nis) and nisoldipine (Nis) by ig 0.3, 1.0, 3.0 and 1.0, 3.0, 9.0 mg.kg-1 respectively on renal hypertensive rats (RHR) and 0.1, 0.3, 1.0 mg.kg-1 orally (for both drugs) on renal hypertensive dogs (RHD) were studied. Both m-Nis and Nis depressed blood pressure (BP) dose-dependently in RHR and RHD. The reduction of blood pressure correlated well with the m-Nis concentration in plasma of RHD. On the basis of ED20 (HR)/ED20 (BP), the hypotensive effect of m-Nis on systolic blood pressure (SBP) was only 1.6 times as great as that of Nis on RHR (P less than 0.05), but in RHD, both drugs showed the same potency (P greater than 0.05). In both models, m-Nis showed much more potent effect on diastolic blood pressure (DBP) than Nis (P less than 0.01), and possessed stronger hypotensive effects on DBP than on SBP (P less than 0.05 and P less than 0.01); but for Nis, its effects on SBP and DBP appeared to be in the same order (P greater than 0.05). The fall in BP was accompanied by a transient increase of heart rates (HR) with m-Nis and Nis in RHR and RHD. The chronic antihypertensive effects of m-Nis and Nis were also remarkable with 1.0 mg.kg-1 daily at 9 AM for 21 d. During this period, the BP and HR lowered to nearly normal level. After withdrawal of m-Nis and Nis, the hypotensive effects lasted nearly 1 wk.

To understand mechanisms of signal transduction involved in the regulation of isotype differentiation of B lymphocytes, we investigated effects of activation of protein kinase C (PKC) by phorbol esters and elevation of intracellular free calcium (Ca2+) by the calcium ionophore ionomycin (Ion) on Ig secretion by mouse Peyer's patch (PP) and spleen B cells. Results show that Ion suppressed production of IgM, IgG, and IgA by LPS-stimulated B cells whereas PKC-activating phorbol esters also inhibited LPS-induced IgM and IgG secretion, but induced a substantial IgA synthesis, as well as alpha-chain mRNA transcription, in B cells whether stimulated or not by LPS. Phorbol esters enhanced IgA response by directly activating PKC, inasmuch as the other phorbol ester, 4 alpha-phorbol 12,13-didecanoate, which is inactive with respect to PKC, had no effect on B cell differentiation. The increase in IgA secretion occurred in whole PP B cells, but not in the membrane IgA- B cell subset, suggesting that PKC activation does not promote the switching rate of IgM+ cells to IgA+ cells. Results from double staining studies of mIgA using FITC-labeled anti-IgA antibodies and DNA content using the DNA-binding propidium iodide showed that enhanced IgA response was not caused by IgA B cell clonal expansion. PMA induced low level of IL-6 production by highly purified PP B cells. However, addition of anti-mouse IL-6 antibody did not prevent PMA-enhanced IgA secretion, suggesting that IL-6 was not responsible for IgA induction by PMA. Collectively, the present data demonstrate that PKC activation and Ca2+ mobilization, which synergistically trigger cell proliferation, have differential effects on B cell isotype differentiation. Elevation of intracellular Ca2+ suppresses Ig production, but activation of PKC selectively enhances IgA secretion by directly promoting terminal differentiation of IgA-committed PP B cells into IgA-secreting plasma cells.