Liguisticum wallichii inhibits renal carcinoma progression by downregulating UBE3A and through suppression of NF-κB signaling.

Renal carcinoma accounts for a fifth of the morbidity among malignant tumors in China. Ubiquitin-protein ligase E3A (UBE3A) plays an important role in the occurrence and development of gene mutation-induced diseases. This study was designed to investigate the mechanism of Liguistium wallichii in treating renal carcinoma. Hematoxylin and eosin staining was applied to detect the pathological changes in a rat renal carcinoma model. The experimental group received L. wallichii treatment at 100 mg/kg every 48 h for 4 weeks, while the control group only received normal saline. The proliferation index Ki67 was measured by immunohistochemistry. Primary renal carcinoma cells were isolated and UBE3A expression was measured by quantitative polymerase chain reaction. The related signaling pathway was screened by the Pathway Finder Array. pP65 nuclear import was detected by immunofluorescence. A total of 60 rats were used for the renal carcinoma model, of which 58 rats were successfully established and equally divided into two groups: L. wallichii and normal saline. Ki67 expression decreased in the L. wallichii group and was upregulated in the normal saline group. Histological analysis showed significant renal cell nucleus division in the normal saline group. The UBE3A level decreased after L. wallichii treatment compared to the level in the normal saline group. The Pathway Finder Array revealed that the NF-κB signaling pathway was activated, and pP65 presented obvious nuclear import in the normal saline group. In conclusion, L. wallichii inhibits renal carcinoma progression by downregulating UBE3A and suppressing the NF-κB signaling pathway.

TGF-ß1 and Serpine 1 expression changes in traumatic deep vein thrombosis.

The objective of this study was to investigate the expression changes of transforming growth factor ß1 (TGF-ß1) and Serpine 1 in rats with traumatic deep vein thrombosis (DVT). In total, 60 male Sprague Dawley rats were divided into model (N = 50) and control groups (Group A, N = 10). From the model group, 10 rats were randomly selected after modeling as the pre-thrombosis group (Group B, N = 10), and the remaining 40 rats in the model group were divided into the thrombosis (Group C) and no thrombosis groups (Group D) depending on whether DVT was apparent at 25 h after modeling. All rats were dissected and the total RNAs of the femoral veins were extracted. TGF-ß1 and Serpine 1 expression was detected by microarray and polymerase chain reaction (PCR) analyses, and the related signal pathways were analyzed using bioinformatic analysis. Of the 40 rats, DVT was evident in 23, yielding an incidence rate of 57.50%. TGF-ß1 and Serpine 1 expression increased significantly at 2.5 h after modeling, while DVT began to form at 25 h after modeling. Both PCR and microarray analysis showed that TGF-ß1 and Serpine 1 expression levels were significantly higher in the thrombosis group than in the other groups (P < 0.05). Bioinformatic analysis indicated that TGF-ß1 was an upstream regulatory gene of Serpine 1 and could induce Serpine 1 overexpression. Together, these results suggested that TGF-ß1 and Serpine 1 overexpression might play an important role in DVT formation and have predictive values.

Production of transgenic kiwifruit plants harboring the SbtCry1Ac gene.

The kiwifruit (Actinidia chinensis Planch.) is an economically and nutritionally important fruit crop that has a remarkably high vitamin C content and is popular throughout the world. However, kiwifruit plants are vulnerable to attack from pests, and effective pest control is urgently required. Transgenic kiwifruit plants containing the synthetic chimeric gene SbtCry1Ac that encodes the insecticidal protein btCrylAc were obtained through an Agrobacterium-mediated transformation of kiwifruit leaf discs. The kanamycin resistance of the transgenic plants was then analyzed. Results from polymerase chain reactions and genomic DNA Southern blot analyses indicated that SbtCrylAc had been integrated into the genomes of these plants. The results of insect bioassays revealed that the average Oraesia excavate inhibition rate of plants tested at 10 days post-infestation was 75.2%. To our knowledge, this is the first study that has developed insect-resistant transgenic kiwifruit plants.

Effects of General-epidural Anaesthesia on Haemodynamics in Patients with Myasthenia Gravis.

OBJECTIVE: The current study aims to explore the effects of general-epidural anaesthesia (GEA) on the perioperative haemodynamics in patients with myasthenia gravis (MG), as well as their extubation time. METHODS: A total of 42 MG patients (Ossermann I-II b types) receiving elective total thymectomy were randomized into GEA (n = 20) and general anaesthesia alone (GA; n = 22) groups. Changes in their mean arterial pressure (MAP) and heart rate (HR) were recorded before anesthesia and at the time of intubation, skin incision, sternotomy and extubation. Dosages of general anaesthetics during time unit and the time of extubation and complete recovery from the ending of the operation were also recorded. RESULTS: After anaesthesia, both groups displayed increased MAPs and HRs, with those in the GA group significantly higher than those in the GEA group (p < 0.05). The total consumption of general anaesthetics in the GA group was markedly higher than that in the GEA group (p < 0.01). CONCLUSION: The GEA group had shorter postoperative extubation and recovery time than the GA group (p < 0.01). General-epidural anaesthesia stabilizes perioperative haemodynamics, reduces the consumption of general anaesthetics and shortens extubation time. It is a feasible and ideal anaesthetic method at present.

Detection of targets and their mechanisms for early diagnosis of traumatic deep vein thrombosis.

The purpose of this investigation was to identify targets for the early diagnosis and predictors of deep venous thrombosis (DVT) and the role of these targets in the formation of venous thrombosis. A model of DVT was constructed in rats. Thromboses and venous walls were sampled for reverse transcription polymerase chain reaction study, and blood was sampled for enzyme-linked immunosorbent assay studies. Vein endothelial cells were cultured to observe the effects of interleukin (IL)-17 on the expression of tissue plasminogen activator (t-PA)/plasminogen activator inhibitor type 1 (PAI-1). IL-17 monoclonal antibody was used to study its effect on preventing the formation of DVT. One-hundred and twenty hours after the animal model was constructed, significant DVT started to form. Polymerase chain reaction tests showed that immediately after the model was created, the expression of IL-17 increased greatly, whereas the balance between t-PA and PAI-1 was disrupted just before DVT formed. The increase of serum IL-17 was positively related with the formation of DVT. Thus, the application of IL-17 monoclonal antibody could reduce the formation of DVT in rats. IL-17 might be a target for the early diagnosis of DVT and should be further studied to assess its clinical value.

Genetic diversity and population genetic structure of the miiuy croaker, Miichthys miiuy, in the East China Sea by microsatellite markers.

Genetic diversity and patterns of population structure of the miiuy croaker were investigated using SSR markers. A set of 10 microsatellite loci revealed 40 alleles; the number of alleles varied from 2 to 10 for each marker. A relatively high level of genetic variability was observed between miiuy croaker individuals. Genetic diversity was relatively high within populations with corresponding high average gene flow. There were genealogical branches or clusters corresponding to sampling localities according to the UPGMA tree and principal component analysis. Knowledge of the genetic diversity and population structure will be crucial for establishing appropriate fishery management stocks for this species.

New roles of filopodia and podosomes in the differentiation and fusion process of osteoclasts.

The cytoskeleton mediates various cellular processes such as differentiation and fusion, including in the filopodia and podosomes. However, apart from cell migration and formation of the sealing zone, little is known regarding the changes and related regulatory mechanisms of the cytoskeleton and additional roles of the filopodia and podosomes during the differentiation and fusion of osteoclasts. The cytomorphology and cytoskeleton of osteoclasts in the differentiation process were evaluated using tartrate-resistant acid phosphatase staining and immunofluorescence staining. Moreover, the expression levels of Rho GTPases and enzymes related to osteoclast differentiation and bone resorption were detected by quantitative reverse transcription-polymerase chain reaction. We detected 3 types of filopodia in osteoclast precursors and only 1 type of filopodia in undifferentiated cells. Mature osteoclasts were completely devoid of filopodia. Interestingly, cell fusion was highly specific, and the fusion initially occurred to the filopodia. Confocal images revealed that F-actin and microtubules significantly differed among fused cells. These results suggest that filopodia and podosomes not only play important roles in cell migration and the formation of sealing zones but also in the pre-fusion selectivity of 2 cells and the movement direction of the cell nucleus and cytoplasm during the fusion process. In addition, cdc42v1, RhoU, and RhoF regulate the formation of 3 types of filopodia during various stages of differentiation, while Rac1, Rac2, and filament A may be associated with cell selectivity during the fusion process.

Isolation and characterization of polymorphic microsatellite markers from Coilia ectenes.

Coilia ectenes (Jordan and Seale 1905) is an important anadromous species that is an important resource at risk of extinction because of over-fishing, pollution, and coastal construction. To evaluate the genetic diversity of C. ectenes for use in breeding programs, elite microsatellite-enriched libraries were constructed and novel microsatellite markers were developed, and applied to genetically detect wild populations. Out of 92 randomly selected and sequenced clones, 89 contained a CA or GA repeat motif. Twenty-two pairs of primers were designed to investigate the polymorphism and genetic structure of a wild population collected from the Yellow River estuary, China. It was found that 2 loci were monomorphic and 20 loci were polymorphic. The number of alleles per polymorphic loci ranged from 3 to 13, with an average of 7.9. The expected heterozygosity per locus ranged from 0.05 to 0.89, with an average of 0.68. The isolated polymorphic markers are expected to be of use in future genetic breeding programs for C. ectenes, and in the assessment of genetic variation within this species.

Isolation of novel microsatellite markers from Paralichthys lethostigma (Paralichthyidae) and their cross-species application in Pleuronectiformes.

We investigated the genetic diversity of the southern flounder Paralichthys lethostigma. Microsatellite-enriched libraries were constructed and novel microsatellite markers were developed and applied for genetic detection of wild populations. Cross-species amplification was also conducted in five pleuronectiforme species. Of 45 randomly selected and sequenced clones, 43 contained a CA or GA repeat motif. Fourteen pairs of primers were designed to investigate the polymorphism and genetic structure of a wild population collected from North Carolina State coastal waters. Two loci were monomorphic and 12 loci were polymorphic. The number of alleles per polymorphic locus ranged from 2 to 16, with an average of 7.3, and the expected heterozygosity per locus ranged from 0.10 to 0.92, with an average of 0.58. Cross-species amplification showed that most of the markers could successfully amplify Paralichthys olivaceus DNAs, few markers amplified in Verasper variegatus and Verasper moseri, and none of them could amplify Scophthatmus maximus and Cynoglossus semilaevis DNAs. The isolated polymorphic markers would be useful for the genetic breeding and assessment of genetic variation within the genus Paralichthys.