Objective To investigate the inhibitory effect of extracts from asparagus filicinus rhizome on prolifieration of human osteosarcoma Saos-2 cells and its molecular mechanism .Methods MTT assay was used to detect the cytotoxic activity and growth inhibition of three different extracts from asparagus filicinus rhizome against Saos-2 cells ;plate colony formation assay was per-formed to detect active fraction of asparagus filicinus rhizome on the anchorage dependent growth of Saos-2 cells ;the cell cycle alter-ation was determined by propidium iodide staining and flow cytometry analysis ;the alteration of protein expression level of COX-2 was determined by using Western blotting .Results Ethyl acetate fraction of asparagus filicinus rhizome (AF-A) exerted the potent cytotoxicity on Saos-2 cells(IC50 =26 .7 μg/mL);AF-A induced the inhibitory effect on the anchorage dependent growth of Saos-2 cells in a dose dependent manner(P<0 .05);Saos-2 cells treated by AF-A at the concentration of 30 .0 and 100 .0 μg/mL for 48 h induced the increase of percentages of S phage from (31 .8 ± 4 .8)% in the control group to (43 .7 ± 2 .5)% and(51 .9 ± 1 .9)% ,the difference showing statistical significance (P< 0 .05) .Western blotting showed that AF-A at different concentrations decreased COX-2 protein expression .Conclusion AF-A posseses the inhibitory effect on the proliferation and growth of human osteosarcoma cells in vitro ,and its mechanism might be associated with the induction of S phage arrest and the inhibition of COX-2 protein ex-pression level .

Objective To investigate the relationship between the expression of the survivin gene and CpC methylation in exon 1 of the survivin gene in CA tissue, and to study the expression of survivin protein in CA tissue and its modulation mechanism. Methods Tissue samples were obtained from the CA lesions of 30 patients, normal cervix of 10 female controls, and normal foreskin of 10 male controls. Immunohistochemistry was carried out to detect the expression of survivin protein in these specimens, RT-PCR to measure the mRNA expression of survivin, and methylation specific PCR (MSP) to analyze the methylation status of CpG island in the survivin gene exon 1. Results The positivity rate of survivin protein and mRNA was 90% (27/30) and 93.3% (28/30) in CA tissue specimens, respectively, 5% (1/20) and 5% (1/20) in control tissue specimens, respectively; there was a significant difference between the two groups of specimens in both the parameters (x2 = 35.187, 38.437, both P < 0.01). The demethylation of CpG island in the survivin gene exon 1 was observed in 86.7% (26/30) of the CA tissue specimens and in 15% (3/20) of the control tissue specimens (x2 = 10.865, P < 0.01). There was a positive correlation between the demethylation status of CpG island in exon 1 and the mRNA expression of survivin gene (x2 = 13.929, P = 0.014). Conclusions The expression of survivin protein in CA tissues might be associated with the demethylation of CpG island in exon 1 of the survivin gene, and may play a certain role in the development of CA.

Age-related macular degeneration(AMD), the most common cause of irreversible blindness over 50 years in developed countries, is a degenerative disease occurs with age. The prevalence rate in our country is also increasing by year and the pathogenesis is still undiscovered. In this review, we focus on the associated risk factors reported in recent years, aim to further understand the pathogenesis of AMD.

Objective: To estimate the burden of disease (BOD) attributable to main risk factors of chronic diseases in Zhejiang Province in 2017, so as to provide the evidence for formulating the control strategy for chronic diseases and reducing BOD. Methods: The results of the Global Burden of Disease Study 2017 ( GBD 2017 ) were extracted to evaluate years of life lost due to premature mortality ( YLL ), years lived with disability ( YLD ) and disability-adjusted life years ( DALY ). The gender- and age-specific BOD attributable to main risk factors of chronic diseases, including the environment, metabolism and behaviors, in Zhejiang Province in 2017 was estimated and compared with those in 1990. Results: High DALY rates of chronic diseases were estimated attributable to tobacco use ( 2 807.08/105 ), unreasonable diet ( 2 724.72/105 ) and hypertension ( 1 878.69/105 ) in Zhejiang Province in 2017, and high DALY rates of chronic diseases were estimated in men attributable to tobacco use ( 4 764.77/105 ), unreasonable diet ( 3 297.00/105 ) and hypertension ( 2 076.92/105 ), while high DALY rates of chronic diseases were estimated in women attributable to unreasonable diet ( 2 117.16/105 ), hypertension ( 1 668.24/105 ) and hyperglycemia ( 1 100.53/105 ), respectively. Among individuals at ages of 15 to 49 years, high DALY rates of chronic diseases were estimated attributable to unreasonable diet ( 759.29/105 ), drug abuse ( 611.71/105 ) and tobacco use ( 605.37/105 ); among individuals at ages of 50 to 69 years, high DALY rates of chronic diseases were estimated attributable to tobacco use ( 5 528.37/105 ), unreasonable diet ( 4 628.18/105 ) and hypertension ( 2 757.78/105 ); and among individuals at ages of 70 years and older, high DALY rates of chronic diseases were estimated attributable to unreasonable diet ( 16 370.09/105 ), tobacco use ( 15 551.40/105 ) and hypertension ( 14 408.63/105 ). As compared to those in 1990, the DALY rates of chronic diseases attributable to high body mass index, alcohol use, hyperglycemia, high low-density lipoprotein cholesterol and drug abuse increased by 108.23%, 48.59%, 23.17%, 17.64% and 6.06%, and the DALY rates of chronic diseases attributable to air pollution, occupational risks, unreasonable diet and impaired renal function reduced by 51.11%, 44.81%, 22.49% and 19.83%, and no significant alterations were detected in DALY rates of chronic diseases attributable to tobacco use or hypertension in 2017. Conclusions There was a high BOD of chronic diseases attributable to tobacco use, unreasonable diet and hypertension in Zhejiang Province in 2017, and the BOD of chronic diseases attributable to high body mass index, alcohol use and hyperglycemia appeared a tendency towards a rise in Zhejiang Province in 2017 relative to in 1990.

Objective To detect the expression and function of cysteinyl leukotriene receptors (CysLTRs)in keratinocytes.Methods Human keratinocytes were isolated from the tissue of foreskin by digestion with dispase Ⅱ and trypsin,and subjected to primary culture.By using confocal laser scanning microscopy and reverse transcriptase PCR,the localization and expression of CysLTRs were studied in kemtinocytes.respectively.Some primarily cultured keratinocytes were pretreated with leukotriene D4 (30 nmoi/L),MK571(300 nmol/L),and BAYu9773 for 5 minutes followed by the detection of intmcellular calcium level using the Ca2+ indicator dye Fura-2/AM as well as cell proliferation bv MTT assay.Results The expressions of CysLTR1 and CysLTR2 were observed in cultured keratinocytes,and they were mainly located on cell membrane,partly in cytoplasm and nuclei.Compared with non.stimulated cells,a significant increase Was noted in the expression of CysLTRs,especially in the nuclei of keratinocytes stimulated by LTD4(P＜0.05),together with an elevation in intracellular calcium level(42.27±3.00 mmol/L,P＜0.01)and acceleration in cell proliferation (P＜0.01).However,both MK571 and BAYu9773 could completely block the effect of LTD4 on intmcellular calcium level and cell proliferation.and there was no significant difference in the blocking effect between MK571 and BAYu9773.Conclusions Functional CysLTRs are expressed in human keratinocytes.and they carl increase the intracellular calcium level in,and cell proliferation of,keratinocytes.

Objective:To investigate the expression of CXCL12-CXCR4 and VEGF-C in pancreatic cancer and relation to clinical pathology.Methods:The tissue samples including PAC,the cancerous peripheral tissues,the normal pancreatic tissues and peripheral lympho nodes were obtained from 30 patients with PAC.The expressions of CXCL12,CXCR4and VEGF-C proteins in these tissues were assayed by immunohistochemical staining.The expressions of CXCL12,CXCR4 and VEGF-C mRNA in PAC were also investigated by fluorescence quantitative real-time PCR.Results:In all the samples,the positive rates of CXCL12 protein in PAC,the cancerous peripheral tissues,the normal pancreatic tissues and peripheral lympho nodes were respectively 13.3%(4/30),46.7%(14/30),56.7%(17/30) and 50.0%(15/30).The positive rates of CXCR4 protein in PAC,the cancerous peripheral tissues,the normal pancreatic tissues and peripheral lympho nodes were respectively 80.0%(24/30),70.0%(21/30),26.7%(8/30) and 73.3%(22/30).The expression levels of CXCR4 mRNA in PAC tissues,the cancerous peripheral tissues and peripheral lympho nodes were higher than that in the normal pancreatic tissues(P

The clinical data of 13 patients diagnosed as Takayasu's arteritis with cardiomyopathy in Peking Union Hospital were reviewed.Echocardiography showed cardiomegaly (left chambers mainly),diffuse weakened myocardial wall movement and reduced left ventricular ejection fraction.Dilated cardiomyopathy was diagnosed in two patients.Glucocorticoid and cyclophosphamide were the essential medication for treatment.One patient died,1 patient was aggravated and the disease condition in remaining 11 patients was stable or improved.Takayasu's arteritis with cardiomyopathy is a rare disease and it attacks younger people.It is difficult to identify at onset,and the disease deteriorates rapidly.Echocardiography should be performed earlier to evaluate the heart function for early diagnosis and treatment.Administration of large dose of glucocorticoid and cyclophosphamide at early stage may improve the prognosis.

Objective To investigate the value of double-balloon endoscopy (DBE) and multi-slice CT enteroclysis (MSCTE) in diagnosis of Crohn's disease (CD) in small intestine. Methods DBE and MSCTE were performed in 71 patients with suspected Crohn's disease in small intestine. The two methods were compared in terms of diagnosis, extents of disease, existance of complications and activity of the disease according to the pathologic findings and the outcome of follow-up. Results The diagnostic yields of DBE and MSCTE were comparable with no significant difference (χ2=2.29, P> 0.05). The positive and negative likelihood ratios were 22.5 and 0. 022 in DBE respectively, and were 1.6 and 0. 240 in MSCTE respectively. The results of DBE was consistent with MSCTE in diagnosis of mild bowel stenosis, but was inconsistent with MSCTE in diagnosis of moderate-severe bowel stenosis (χ2=11.298, P=0.001). The concordance of two methods in diagnosis of disease activity was 95.8%. Conclusions The first choice in diagnosis of small bowel CD is DBE. The combination of two methods will be helpful in diagnosis and evaluation of CD severity.

The recombinant adenoviruses containing pGH cDNA and pGH cDNA with the first intron under the control of CMV promoter were constructed respectively by homogenous recombination method. The results showed that the recombinant adenoviruses could mediate pGH cDNA expression in CHO cells infected with the recombinant adenoviruses. The expression level of pGH cDNA with the first intron increased by 117% compared with pGH cDNA without intron. This indicate that the first intron of pGH gene have the function of improving the expression of the pGH gene.
Adenoviridae ; genetics ; Animals ; CHO Cells ; Cricetinae ; Cytomegalovirus ; genetics ; Gene Expression ; Genetic Engineering ; Genetic Vectors ; genetics ; Growth Hormone ; genetics ; Introns ; Promoter Regions, Genetic ; Recombinant Fusion Proteins ; genetics ; Swine

Background Human limbal allograft transplantation or limbal autograft transplantation are the primary approaches to the severe corneal-blindness,but their application in clinic were limited because of the defects of donor material.With the development of tissue engineering technology,transplantation of in vitro cultured limbal epithelial stem cells is being an advanced management.Objective The aim of this work was to expand human limbal epithelial stem cells ex vivo under the guidance of confocal microscope and to lay the foundation for fabricating ex vivo cultured cell sheets.Methods Ten eyes of ten patients were examined with the Heidelberg Retina Tomography Ⅲ Rostock Cornea Module(HRT3-RCM)to elucidate the structure of the human corneoscleral limbus and to correlate limbal epithelial dimensions.According to the analysis of the images of limbal epithelia,the limbal tissues provided by Eye Bank of Henan Eye Institute were cut into suitable explants.Then,this study was conducted to expand limbal epithelial stem cells ex vivo on denuded amniotic membrane.The phenotypes of primary cultured cells were evaluated by morphology and immunofluorescent staining with antibodies for limbal epithelial stem cell markers (p63,cytokeratinl9)and differentiation markers(keratin 3,involucrin).This experimental procedure was approved by the Ethic Committee of Henan Provincial People's Hospital.The written informed consent was obtained from subjects before initiation of any examination.Results The palisade morphology of human limbus was imaged clearly on the laser scanning in vivo confocal microscopy and many hyperreflective cells were observed in palisade basal cells.The cell-island phenomenon was seen in the basement membrane under the laser scanning in vivo confocal microscopy.The oblique sections of limbus showed many papilla-like epithelial columns below the superficial limbal epithelia.Throughout the experiment duration,the epithelial cells grew well with the migration rates from limbal tissue (68.62± 16.94)％ and the migration time(5.83 ±2.04)days,which depended on the tissue freshness.Compared with the second and forth batch of tissue,the migration rates of the third and sixth batch of tissues were significantly higher(P＜0.05),and the migration time was evidently longer in the forth and sixth batch of tissue compared with the first,second,third and fifth batch(P＜0.05).The positively expressing rates in the cultured corneal stem cells were 4.05％ and 36.52％ for p63,26.07％ and 40.55％ for CK19,57.88％ and 40.81％ for K3,64.66％ and 59.19％ for involucrin.Conclusion Human limbal epithelial stem cells can be successfully and purposefully obtained from the limbal tissue based on the guidance confocal miscroscope.The cultured corneal stem cells can grow well on the denuded amniotic membrane