Cornuside ameliorated experimental autoimmune encephalomyelitis by limiting the recruitment of CD4+ T lymphocytes in the spinal cord

We conducted this study to determine whether cornuside could improve the neurological deficit symptoms of experimental autoimmune encephalomyelitis (EAE) rats, as well as determine the potential involvement of CD4+ T lymphocytes, vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and tumor necrosis factor-α (TNF-α). Altogether, 32 Lewis rats were randomly divided into control, EAE, EAE/prednisolone, and EAE/cornuside, wherein their neurological function was assessed every day. CD4+ T lymphocyte recruitment into the spinal cord (SC) was evaluated using immunohistochemistry. The VCAM-1, ICAM-1 and TNF-α mRNA expressions in the SC were determined by real-time quantitative PCR, and the VCAM-1 and ICAM-1 proteins were determined by western blotting. Compared to the control group, the EAE group rats with neurological deficits had enhanced CD4+ T lymphocyte infiltration and higher expression levels of VCAM-1, ICAM-1, and TNF-α in the SC. Meanwhile, compared with the EAE group, the EAE/cornuside and EAE/prednisolone groups had lower neurological scores, less CD4+ T lymphocyte infiltrations, and lower expression levels of VCAM-1, ICAM-1, and TNF-α in the SC. Thus, cornuside ameliorated EAE, which could be owed to the inhibition of CD4+ T lymphocyte recruitment and VCAM-1, ICAM-1, and TNF-α expressions in the SC

Preliminary study on the role of novel LysR family gene kp05372 in Klebsiella pneumoniae of forest musk deer / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

LysR-type transcriptional regulators are involved in the regulation of numerous cellular metabolic processes in Klebsiella pneumoniae, leading to severe infection. Earlier, we found a novel LysR family gene, named kp05372, in a strain of K. pneumoniae (designated GPKP) isolated from forest musk deer. To study the function of this gene in relation to the biological characteristics of GPKP, we used the suicide plasmid and conjugative transfer methods to construct deletion mutant strain GPKP-Δkp05372; moreover, we also constructed the GPKP-Δkp05372+ complemented strain. The role of this gene was determined by comparing the following characteristics of three strains: growth curves, biofilm formation, drug resistance, stress resistance, median lethal dose (LD50), organ colonization ability, and the histopathology of GPKP. Real-time polymerase chain reaction (RT-PCR) was used to test the expression level of seven genes upstream of kp05372. There was no significant difference in the growth rates when comparing the three bacterial strains, and no significant difference was recorded at different osmotic pressures, temperatures, salt contents, or hydrogen peroxide concentrations. The GPKP-Δkp05372 mutant formed a weak biofilm, and the other two strains formed medium biofilm. The drug resistance of the GPKP-Δkp05372 mutant toward cephalothin, cotrimoxazole, and polymyxin B was changed. The acid tolerance of the deletion strain was stronger than that of the other two strains. The LD50 values of the wild-type and complemented strains were 174-fold and 77-fold higher than that of the GPKP-Δkp05372 mutant, respectively. The colonization ability of the GPKP-Δkp05372 mutant in the heart, liver, spleen, kidney, and intestine was the weakest. The three strains caused different histopathological changes in the liver and lungs. In the GPKP-Δkp05372 mutant, the relative expression levels of kp05374 and kp05379 were increased to 1.32-fold and 1.42-fold, respectively, while the level of kp05378 was decreased by 42%. Overall, the deletion of kp05372 gene leads to changes in the following: drug resistance and acid tolerance; decreases in virulence, biofilm formation, and colonization ability of GPKP; and regulation of the upstream region of adjacent genes.

Autotoxicity in Pogostemon cablin and their allelochemicals

Abstract The effects of allelochemicals and aqueous extracts from different Pogostemon cablin (Blanco) Benth., Lamiaceae, parts and rhizosphere soil on growth parameters, leaf membrane peroxidation and leaf antioxidant enzymes were investigated in patchouli. P. cablin seedlings were incubated in solutions containing allelochemicals and aqueous extracts from different patchouli parts and its rhizosphere soil at several concentrations. Firstly, the growth parameters were significantly reduced by the highest concentration of leaves, roots and stems extracts (p < 0.05). As compared to the control, plant height was reduced by 99.8% in the treatment with leaves extracts (1:10). The malondialdehyde content increased greatly when patchouli seedlings were subject to different concentrations of leaves, roots and stems extracts; meanwhile, the superoxide dismutase and peroxidase activities showed an increase trend at the low concentration, followed by a decline phase at the high concentration of roots and leaves extracts (1:10). What's more, leaves and roots extracts had a more negative effect on patchouli growth than stems extracts at the same concentrations. Secondly, the total fresh mass, root length and plant height were greatly reduced by the highest strength of soil extracts. Their decrements were 22.7, 74.9, and 33.1%, respectively. Thirdly, growth parameters and enzymatic activities varied considerably with the kinds of allelochemicals and with the different concentrations. Plant height, root length and total fresh weight of patchouli were greatly reduced by p-hydroxybenzoic acid (200 μM), and their decrements were 77.0, 42.0 and 70.0%, respectively. Finally, three useful measures on reducing the autotoxicity during the sustainable patchouli production were proposed.

Dynamic accumulation of sesquiterpenes in essential oil of Pogostemon cablin

Sesquiterpenes Essential oil produced by patchouli was one of the most important naturally occurring base materials used in the perfume industry, containing various sesquiterpenes. Three different parts (leaves, stems and roots) of Pogostemon cablin (Blanco) Benth., Lamiaceae, were profiled in relation to different maturation phases in this paper, evaluating the variations in content of the major sesquiterpenes in the essential oil. Twelve sesquiterpenes were analyzed by GC-MS throughout the maturity of P. cablin. Patchouli alcohol (37.54%-51.02% in leaves, 28.24%-41.96% in stems and 14.55%-35.12% in roots) was the major sesquiterpene during the maturation of the plant. The average content of several other sesquiterpenes (α-bulnesene, α-guaiene, seychellene, β-humulene and caryophyllene) were higher than 3% among leaves, stems and roots. The content of essential oil, patchouli alcohol, α-bulnesene and several other compounds were highly accumulated at 210 days of maturation after cultivation of P. cablin. Thus, this period was the best moment to exploit the maximum level of these high value-added compounds in P. cablin. Furthermore, our results indicated that the essential oil extracted from leaves of P. cablin has the highest potential to be used in the perfume industry.

The prognostic impact of O 6- methylguanine DNA methyltransferase and epidermal growth factor receptor expressions on primary gliosarcoma: A clinicopathologic and immunohistochemical study of seven cases at a single institution.

Context: Gliosarcoma is an uncommon variant of glioblastoma characterized by a biphasic tissue pattern of glial and mesenchymal differentiation. O 6- methylguanine DNA methyltransferase (MGMT) is a DNA repair protein that removes mutagenic and cytotoxic adducts from O 6 -guanine in DNA. Lack of MGMT protein expression immunohistochemically is related to drug responses in patients of malignant glioma treated with alkylating agents. Epidermal growth factor receptor (EGFR) is the most frequently amplified gene in glioblastoma and associated with tumor invasiveness, angiogenesis, poor survival, and resistance to radiation therapy. Aims: To elucidate the relationship between the statuses of the MGMT as well as EGFR proteins and the prognosis. The study was undertaken on samples received at the Department of Pathology from 2003 to 2009. Materials and Methods: Clinicopathologic and immunohistochemical study of seven cases was performed. Results: This series included three men and four women with a mean age of 49.3 years at first surgery. The median progression-free survival (PFS) was 22.2 months and 8.6 months for primary tumors with 0 to 1+ and 2+ to 3+ MGMT staining, respectively; the median overall survival (OS) was 27.5 months and 14.2 months for primary tumors with 0 to 1+ and 2+ to 3+ MGMT staining, respectively. The median PFS was 17.2 months and 11.2 months for primary tumors with 0 to 1+ and 2+ to 3+ EGFR staining, respectively; the median OS was 20.4 months and 17.7 months for primary tumors with 0 to 1+ and 2+ to 3+ EGFR staining, respectively. Conclusions: The series showed that MGMT and EGFR protein expressions were both unfavorable prognostic factors for patients with gliosarcoma.

T cell receptor constant alpha chain gene +1592C/T polymorphism may not be associated with IgA nephropathy / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the relationship between T cell receptor constant alpha chain (TCRCα) gene +1592C/T polymorphism and IgA nephropathy.</p><p><b>METHODS</b>TCRCα +1592C/T genotypes were identified by PCR-RFLP and direct sequencing in 244 Chinese Han patients with IgA nephropathy, who were classified according to their genotype into CC (188 cases), CT (54 cases) and TT (2 cases) groups. The clinical and pathological data of the patients were analyzed in relation to the TCRCα +1592C/T genotypes.</p><p><b>RESULTS</b>No significant differences in the clinical and biochemical indices were found in these patients with different TCRCα gene +1592C/T genotypes. TCRCα +1592C/T polymorphism was not found to contribute to severity or manifestations of renal pathology.</p><p><b>CONCLUSIONS</b>TCRCα+1592C/T polymorphism may not be associated with the susceptibility to IgA nephropathy in the Chinese Han population.</p>

Construction and identification of the incompetent-replication adenovirus carrying the fusion gene composed of prepropeptide of mouse nerve growth factor and human beta-endorphin gene / 中华麻醉学杂志

Objective To construct and identify the incompetent-replication adenovirus carrying the fusiongene composed of the encoding gene of prepropeptide of mouse nerve growth factor(PN)and human beta-endorphin(?-EP)geue.Methods The gene segments of PN obtained from total RNA of the submandibular glandof a 2-week old Kumning mouse were amplified by RT-PCR and joined with the segment of ?-EP to form the fusiongene which was sequenced.The fusion gene contained in the incompetent-replication adenovirus was formed in theBJ-Ad Easy-1 susceptible cells and identified by PCR so as to choose the positive clone without wild vectors.Thecorrect clone was amplified and purified.The titers of adenovirus were determined using the specific 50% tissueculture infection dosage(TCID 50)method.Three days after the adenovirns was transferred into the cultured A431cells,RT-PCR was performed to showed the transcribed mRNA of this fusion gene and the intracellular ?-EPexpression was quanlitatively detected by inummo-histological method.Finally the concentration of human ?-EP inthe culture medium was determined by quantitative radio-immunoassay on 1st,3rd and 7th day afterinfeetion.Results The sequence of the fusion gene was correct.The titer of recombinant adenovirus Ad-NEP was1.5?10~10 pfu/ml.Three days after infection a 475 bp segment was amplified by RT-PCR and abundant orangegranules were shown in the infected cell.The ?-EP concentration in the culture medium was significantly higher inAd-NEP group than in the control group on 1st,3rd and 7th day(P