Objective To investigate the effect of nitrate on acute experimental colitis in mice.Methods A total of 40 BALB/c mice were evenly divided into model group and treatment group.Model group were fed with 4％ dextran sulfate sodium (DSS) solution and treatment group were given 4％ DSS solution and nitrate (1.5 g/L) for seven days.The disease activity index (DAI) of mice was scored.The colon tissue of mice was taken for hematoxylin-eosin staining and myeloperoxidase (MPO)immunohistochemical staining observation.The MPO and activity of nitric oxide in colon tissue were measured by MPO and nitric oxide detecting kit.The data were analyzed by t test.Results At the 6th day and 7th day,the difference of DAI between treatment group and model group was statistically significant (t=5.12 and 6.72,P=0.012 and 0.008).At the 7th day,the tissue score of model group (2.5±0.5) was higher than that of treatment group (1.9±0.4) and the difference was statistically significant (t=3.82,P＜ 0.01).Compared with model group,the histopathological injury of colon tissue in treatment group mice significantly reduced and neutrophil infiltration also decreased.At the 7th day,the concentration of MPO,NO2-and NO3-of model group was (2.8±0.6) U/g,(10.4±4.3) mmol/g and (100.3±50.1) mmol/g respectively,treatment group was (1.5±0.3) U/g,(17.5±7.0) mmol/g and (190.7 ±85.3) mmol/g respectively.The differences were statistically significant (t=11.23,3.81 and 4.50,all P＜0.01).Conclusion Nitrate can reduce DSS-induced acute experimental colitis in mice.

Graphite ; Humans ; Spectrophotometry, Atomic ; methods ; Vanadium ; urine

Objective To construct a cDNA library of three-year old Polygonum multiflorum leaf tissues so as to further research the gene regulation of secondary metabolite biosynthesis of medicinal plants. Methods Total RNA from leaf tissues of P.multiflorum was extracted and mRNA was purified,which were synthesized to double strand cDNA through reverse transcription.After the cDNA termini was blunted,the 5' end of EcoR Ⅰ adapters phosphorylated was conjoined,and then digested by Xho Ⅰ,cDNA fragments were fractionated by Sepharose CL-2B spin column.The fragments longer than 400 bp were linked to Uni-ZAP XR vector.The primary cDNA library was established after the recombinants had been packaged.Uni-ZAP XR Vector might fleetly release pBluescript SK-phasmids at the presence of ExAssist helper phage of coinfection and inverted E.coli SOLR.Finally,PCR and double enzymes digestion were used to analyze the range of inserts,respectively. Results The titer of cDNA primary library was 1.07?106pfu/mL and the length of exogenous insert was at about 0.5-2.0 kb with 5.4?105 recombinants,the recombinants of amplified library were 4.25?1011 and the rate of recombination was 98.5%. Conclusion The results indicate that the cDNA library of P.multiflorum leaf tissues has enough volume for screening the desired genes and sets up a basis for studying on gene regulation of secondary metabolite biosynthesis of medicinal plants besides.

Objective To measure the expression of interleukin-13 (IL-13) and its receptors in mycosis fungoides (MF) lesions,and to investigate their clinical significance.Methods A total of 34 paraffin-embedded specimens of MF,which was confirmed by clinical and histopathological features,immunophenotyping and/or T-cell receptor gene rearrangements,were collected from Hangzhou Third People's Hospital between January 2010 and March 2016.According to the tumor-node-metastasis (TNM) staging system,5 patients were at stage I A,9 at stage Ⅰ B,17 at stage Ⅱ A,and 3 at stage Ⅱ B.Ten normal skin tissue specimens served as controls.Immunohistochemical study was conducted to measure the expression of IL-13,IL-13Rα1 and IL-13Rα2.Results IL-13,IL-13Rα1 and IL-13Rα2 were all expressed in atypical lymphoid cells and epidermotropic lymphoid cells in MF lesions at various stages.IL-13Rα2 was highly expressed in all the MF lesions.None of IL-13 and its receptors were expressed in normal skin tissues and lymphocytes.The expression rates of IL-13 and its receptors in MF lesions increased along with the progression of MF.Additionally,the expression rates of IL-13 (10.00％ ± 3.14％),IL-13Rα1 (21.43％ ± 6.88％) and IL-13Ro2 (31.14％ ± 6.38％) significantly decreased in MF lesions at stage Ⅰ compared with those at stage Ⅱ (27.50％ ± 11.00％,39.45％ ± 9.43％,44.40％ ± 11.15％,respectively,all P ＜ 0.05),but no significant differences were observed between stage Ⅰ A and Ⅰ B,or between stage Ⅱ A and Ⅱ B (P ＞ 0.05).Conclusion IL-13 and its receptors,especially IL-13Rα2,may be expected to serve as biomarkers for early diagnosis of MF and prediction of its biological behaviors.

OBJECTIVE To investigate the inhibiting effect of Pingyangmycin on cultured vein endothelial cells in vitro and illustrate the mechanism of Pingyangmycin on treatment cavernous hemangioma of head and neck. METHODS By MTT assay,the inhibiting rates of cultured Ecv-304 cells which were managed with Pingyangmycin for 24h and 48h were compared respectively. The changes in the cell cycle,apoptosis,and Caspase 3 protein expression of the cells managed with Pingyangcin for 24h were examined by flow cytometry(FCM). RESULTS The results showed that the inhibiting rate of Ecv-304 was dependent on the concentration of Pingyangmycin. However,in the same concentration,the inhibiting effect for 24h was stronger than that for 48h. The percentage of G1 phase cells increased while the S phase decreased,but the percentage of G2 phase cells remained unchanged after managed with Pingyangcin for 24h. An apoptosis peak appeared before G1 phase. The apoptosis rate was also concentration-dependant. The expression of caspase 3 increased in every group of different concentration. CONCLUSION Pingyangmycin can possibly arrest the growth of Ecv-304 in the G1 phase through the mechanism of inducing apoptosis. Caspase 3 may play an important role in the induced apoptosis.

Objective To report 2 cases of Epstein-Barr virus-associated cutaneous lymphoproliferative disease (LPD) and to evaluate their relationship with chronic active Epstein-Barr virus (CAEBV) infection.Methods The clinical data on, laboratory examination findings in, treatment and therapeutic response of 2cases of LPD were analyzed. Results Both the patients had chronic intermittent fever, lymphadenopathy, recurrent lesions including papules, papulovesicles, necrosis and variola-like scar in light-exposed and unexposed areas. Pathologically, there was a dermal infiltrate of pleomorphic lymphoid cells with the involvement of perivascular area and some subcutaneous tissue. Immunohistochemical staining showed that most of the infiltrating lymphoid cells were positive for CD8, PCR revealed no TCR-γ gene rearrangement, and in situ hybridization for EBV was positive. The copy of EBV DNA was above the normal range in the peripheral blood from both patients. Clinical status was improved after glucocorticoid treatment. Conclusion The biologic behavior of LPD appears to be a chronic and indolent course and is closely associated with CAEBV.

Aortic valve calcification (AVC) is a pathological process correlated with multiple disease causes and actively regulated by cardiac valve cells. In this study, porcine aortic valve myofibroblasts cultured in vitro were treated with 50 μg z L(-1) of pathological factor tumor necrosis factor α (TNF-α). Tanshinone II A (TSN) with the concentration of 50 mg x L(-1) and TNF-α were combined in incubating cells for 72 h (3 d) and 120 h (5 d). The Western blotting and Real-time PCR were adopted to detect the changes in smooth muscle α actin (α-SMA), bone morphogenetic protein 2 ( BMP2), alkaline phosphatase (ALP) in cells, and expressions of key effect proteins GSK-3β and β-catenin on Wnt/β-catenin signal pathway. According to the findings, TNF-α can significantly increase the expression of myofibroblasts α-SMA and add the transformation activity to them, with nearly no expression of BMP2, ALP and mRNA in the control group and the TSN group but significant increase in their expressions in the TNF-α group (P < 0.01), which showed osteoblast-like phenotype. Moreover, TNF-α down-regulated the expression of up-streaming regulator GSK-3β and mRNA expression (P < 0. 01) , notably increased the expression of key effect protein β-catenin, but with no significant difference in mRNA with the control group and the TSN group. The result demonstrated that TSN showed a certain inhibitory effect on TNF-α's pathological impact (P < 0.05) in a time-dependent manner. Inflammatory factor TNF-α may promote the transformation of aortic valvular myofibroblasts to osteoblast-like phenotype by activating Wnt/β-catenin signal pathway in aortic valvular myofibroblasts, so as to cause AVC. Tanshinone II A can have a preventive effect in AVC by activating GSK-3β proteins and regulating signal transduction of Wnt/β-catenin signal pathway.
Animals ; Aortic Valve ; cytology ; drug effects ; metabolism ; Cells, Cultured ; Diterpenes, Abietane ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Glycogen Synthase Kinase 3 ; genetics ; metabolism ; Glycogen Synthase Kinase 3 beta ; Myofibroblasts ; cytology ; drug effects ; metabolism ; Osteoblasts ; cytology ; drug effects ; metabolism ; Swine ; Tumor Necrosis Factor-alpha ; genetics ; metabolism ; beta Catenin ; genetics ; metabolism

This paper aims to establish a method for the determination of sulfur dioxide in sulfur fumigation Chinese herbs. Sample powder and hydrochloric acid solution were isolated by paraffin layer in order to avoid early reactions, with the generation of sulfur dioxide, headspace with airtight needle was used to transfer sulfur dioxide into gas chromatograph, and detected with thermal conductivity detector. The analytical performance was demonstrated by the analysis of 12 herbs, spiked at four concentration levels. In general, the recoveries ranging from 70% to 110%, with relative standard deviations (RSDs) within 15%, were obtained. The limit of detection (LOD) was below 10 mg x kg(-1). Standard addition can be used for low recovery samples. The method is simple, less time-consuming, specific and sensitive. Methods comparison revealed that gas chromatography is better than traditional titration in terms of method operability, accuracy and specificity, showing good application value.
Chromatography, Gas ; methods ; Fumigation ; Limit of Detection ; Plants, Medicinal ; chemistry ; Sulfur ; chemistry ; Sulfur Dioxide ; analysis

Objective To assess the clinical features and diagnostic index of progressive macular hypomelanosis(PMH).Methods Eight patients with PMH were recruited into this study.Wood's lamp and in vivo confocal laser scanning microscopy(CLSM)were utilized to observe the lesions of all patients.Microbiological culture of lesion specimens from 2 patients was performed.Tissue specimens from 4 patients underwent immunohistochemieal staining with anti-S-100 and anti-TRP-1 antibodies for the detection of melanocyte quantity.Electron microscopy wag utilized to observe ultrastructural features of lesions.Primary culture of melanocytes was carried out with lesional epidermis.Resnits Under Wood's lamp.the lesions of PMH showed punctiform red fluorescence.CLSM revealed complete pigmented tings in lesions with decreased melanin granules compamd with those surrounding normal skin.Microbiological culture grew red fluorescence-producing,gram-positive bacillus which was identified as Propionibacterium acnes.Immunohistochemistry showed no significant difference in the number of S-100-postive cells or TRP-1-positive cells per high power field (× 400)between lesions and surrounding normal skin (8.25±0.96 vs 8.75±1.71,4.25±0.96 vs 4.50±1.29,both P>0.05).Ultrastructural studies showed a large reduction in the number of type Ⅳ melanosomes in lesions of PMH,along with numemus membrane bound bodies and clusteredly distributed,small type Ⅱ-Ⅳ melanosomes.Melanocytes,with morphological similarity to normal melanocytes,were successfully isolated from the lesional tissue,cuhured and identified.Conclusion A primary diagnostic criteria is pro-posed for PMH according to the clinical and experimental studies.

Objective To explore the clinical characteristics and treatment of nodular panniculitis disease in children. Methods Clinical data of 13 cases with nodular panniculitis disease were reviewed retrospectively. Their etiology,clinical manifestation,misdiagnosis cause,pathologic characteristics, treatment and outcome were analyzed. Results Its clinical manifestation was multiform and showed mainly as fever and hypodermic nodule. Concomitant damages to digestive, respiratory, circulatory and renal system might occur in those children with the system type of this disorder. Conclusion Pediatric nodular panniculitis disease can be easily misdiagnosed and lack of specificity in the early stage, and complicates multiple organs damage.