Objective To construct a suppression subtractive hybridization (SSH) library of human renal cell carcinoma. Methods Poly A + RNA was isolated from human renal cell carcinoma (RCC) tissues and matched normal kidney tissues, respectively. Then double strand cDNA were synthesized and restricted by Rsa I.RCC cDNA were divided into two groups and ligated with either adaptor 1 or adaptor 2.After RCC cDNA hybridized with normal kidney cDNA twice and underwent nested PCR,the PCR products were cloned into pT Adv vector and transformed E.coli TOP10F′.Some positive clones randomly picked up were digested and sequenced. Results The SSH library contained 414 positive clones. Random analysis of 265 clones with enzyme restriction showed that 246 clones contained cDNA fragments which were distributed mainly between 300～900 bp. Among 40 arbitrary clones derived from the above 246 clones, No.170 clone is a previously unknown sequence and the other 39 clones were derived from 35 known genes. Conclusions The quality of the SSH library of human RCC is reliable and its construction would lay the foundation for further screening differentially expressed genes in RCC.

Background Retinal hypoxia is one of primary causes of retinal neovascularization,and its mechanism is research hot topic.Studies showed that hypoxia stimulates the activation of many trancripation factors and vascular endothelial cells,which leads to angiogenesis.Besides to the vascular endothelial growth factor,the effect of adenosine on angiogenesis is increasingly concerned.Objective This study was to invastigate the relationship of biological behaviour of human microvascular endothelial cells (HMEC-1) under the hypoxia and adenosine,and to explore the effect of adenosine on angiogenesis under hypoxia.Methods HMEC-1 cell line was cultured in vitro,and the cells were divided into normoxia group and hypoxia group.The cells in the normoxia group were cultured under the 5％ CO2 environment,and those in the hypoxia group were cultured under the 1％ O2,94％ N2 and 5％ CO2 environment.The proliferation ability and percentage of the cells were assayed by cell counting kit-8 (CCK8) and EDU.The migration number and invasive number of the cells were detected by transwell chamber.The expressions of CD39 and CD73 proteins in the cells were tested by Western blot and immunofluorescence technique,and the level of adenosine was measured by high performance liquid chromatography (HPLC).Results The proliferation values (absobancy) were 0.715-±0.067 and 0.821 ±0.056 in the normoxia group and the hypoxia group in 12 hours after culture,and those in 24 hours were 0.946±0.028 and 0.998±0.028,showing significant increase in the hypoxia group compared with the normoxia group (t12h =3.805,t24h =3.222,all at P ＜ 0.01).The precentage of the proliferation in the hypoxia group was evidently higher than that in the normoxia group (t =-6.868,P＜0.01).The number of the cell migration and invasion in 24 hours after culture was 185.3 ± 10.594 and 74.2± 10.741 respectively in the normoxia group,and that in the hypoxia group was 300.7±22.853 and 107.5±7.007,with significant differences between the two groups (t=-12.124,-6.367,both at P＜0.01).The expression levels of CD39 and CD73 proteins in the hypoxia group were significantly higher than those in the normoxia group in bothl2 hours and 24 hours after culture(all at P＜0.05),and the adenosine content in the cells is gradually increased in 2,6,12,24 and 36 hours after culture,with the highest content in 36 hours.The adenosine content was significantly higher in the hypoxia group than in the normoxia group at various time points (t2h =2.469,P =0.017;t6h =5.442,P＜0.001;t12h =3.841,P＜0.001;t24h =4.458,P＜0.001;t36h =2.757,P =0.008;t48h =3.319,P =0.002).Conclusions Compared with the normoxia group,the proliferation,migration and invasion abilities of HMEC-1 are stronger,meanwhile,the expression of CD39 and CD73 as well as the adenosine level in the cells are all increased under the hypoxic condition.It is suggested that the activation of human microvascular endothelial cells might be significantly related to the level of adenosine and its key enzymes.

Objective To study the relationship between clinical diagnosis and children ventricular premature beat(VPB) electrocardiogram location and morph.Methods Both organic heart disease and without organic heart disease relationship with 109 cases of children ventricular premature beat electrocardiogram location and morph were retrospectively analyzed.Results Children ventricular premature beat location shows that organic heart disease mostly results from left ventricle, without organic heart disease often comes from right ventricle. There was significant difference between above two groups (? 2=37.25 P

OBJECTIVE: To investigate the changes of laryngeal cancer Hep-2 cells to cisplatin chemosensitivity after the interference of siRNA of β-catenin gene expression. METHOD: Using a small interference RNA (siRNA) technology interfere β-catenin gene of Hep-2 cells . The mRNA and protein levels of β-catenin in the Hep-2 cells of different groups were detected by qPCR and Western blot. It was divided into siRNA-β-catenin-Hep-2 siRNA group, β-catenin-Neg negative control group and blank control group. Cell proliferation inhibition rate of different concentrations of cisplatin on three groups was detected by MTT assay. Calculate the 50% inhibitory effective concentration IC50 value. Check the change of three groups of cells' apoptosis rate by flow cytometry after the same concentrations of cisplatin stimulation. RESULT: β-catenin-siRNA interference fragment can specifically reduce the expression levels of β-catenin mRNA and protein. qPCR illustrated the expression of mRNA in β-catenin-siR-NA-Hep-2 interference group decreased 70% (P < 0.05) compared with the control group, Western blot results showed that the β-catenin protein expression of interference group (0. 545 ± 0.111) decreased significantly compared with blank control group (1.507 ± 0.139) and negative control group (1.429 ± 0.089), P < 0.05. The IC50 calculation software showed that IC50 of cisplatin on β-catenin-siRNA IC50 interference group is (5.81 ± 0.46)μg/ml, the blank control group is (10.10 ± 1.01) μg/ml, the difference between the two groups has statistical signifi- cance (P < 0.01). Cell apoptosis rate of β-catenin-siRNA interference group was (26.15 ± 0.60)%, significantly higher than the control group (14.16 ± 0.05)%, P < 0.05. CONCLUSION To interfere the expression of β-catenin can effectively enhance the sensitivity of laryngeal cancer cells to chemotherapeutic drugs cisplatin. It provides a theoretical support for the reduction of laryngeal cancer chemotherapy drug cisplatin dosage.
Apoptosis ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; Humans ; Laryngeal Neoplasms ; genetics ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; beta Catenin ; genetics

Objective To investigate the effects of Morroniside on H2O2-induced oxidative injury in SH-SY5Y neuroblastoma cells.MethodsSH-SY5Y cells were pre-incubated with Morroniside(1,10,and 100 μmol/L)for 24 h prior to exposure to H2O2(500 μmol/L)for 18 h.The content of reactive oxygen species(ROS),nitric oxide(NO),glutathione(GSH)were determined.ResultsPretreatment the cells with Morroniside(10 and 100 μmol/L)lowered H2O2-induced ROS release by 37%(P<0.01)and 27%(P<0.05)versus H2O2,and NO release in a dose-dependent manner by 31%(P<0.05),53%(P<0.01)versus H2O2.Morroniside(1,10,and 100 μmol/L)increased GSH level in cells treated with H2O2 in comparison with cells exposed only to H2O2,respectively.ConclusionMorroniside shows neuroprotection effect against H2O2-induced oxidation injury in SH-SY5Y cell.

Neural stem cells are a category of cells with self-renewal and differentiation potential. Proliferation and differentiation of neural stem cells have closely relation with growth factor, cell factor, environmental signal, and so on. In recent years, evidence indicates that neural stem cells possessing multipotency naturally occur in adult brain. The found of adult neural stem cells have changed the theory that central nervous system cannot regenerative. Chinese Traditional Medicine can effectively induce adult neural stem cells to proliferate and differentiate, opening up unprecedented direction in the research for the treatment of nerve regeneration, and showing great potency for curing cerebral ischemic injury.

Astragalus adsurgens seed is one of the most common adulterants of Astragali Complanati Semen in the market, whose morphological characteristics are very similar with A. complanatus seeds (Astragali Complanati Semen). Many identification methods have been reported, such as morphological identification, fluorescence method, ultraviolet spectrophotometry, TLC, HPLC, protein electrophoresis and so on, but there's no much about microscopic identification. In the present study, the morphological characteristics and microscopic characteristics of these two seeds were investigated, which could provide scientific evidence for the identification and classification of Astragali Complanati Semen. Our results showed that these two seeds were slightly different in the color and the appearance, but significantly different in the microstructure of the seed coat and the hilum, and the distribution of fat droplets in the cotyledon cells. So these microscopic characteristics can be applied for the identification of Astragali Complanati Semen.
Astragalus Plant ; anatomy & histology ; chemistry ; Color ; Discriminant Analysis ; Microscopy ; methods ; Quality Control ; Seeds ; anatomy & histology ; chemistry

Using in vitro everted gut to investigate the intestinal absorption of the extracts from Polygonum orientale at different concentration. UPLC-MS/MS was used to detect the content of protocatechuic acid, isoorientin, orientin, vitexin, cynaroside, quercitrin, kaempferol-rhamnoside in different intestinal segments, then compared the results with the absorption of chemical components of extractive P. orientale in each intestinal segments, and calculated the absorption parameter. We took the statistic analysis with SPSS statistic software. The influence significance of each factors were analyzed to describe the character of absorption. The absorption of each component is linearity in different intestinal segments and different dose, and the square of coeficient correlation exceed 0.95, which consistent with zero order rate process. The K(a) increase along with the raised dosage of the extractive P. orientale (R2 > 0.95), indicated it is the passive absorption; different intestinal segments have different absorption. And the absorption trend in intestinal is duodenum, jejunum, ileum are greater than the colon. As ingredients are selectively absorbed in intestinal sac, the everted intestinal sac method is selected to assess the intestinal absorption charcteristics of ingredients of extractive P. orientale.
Animals ; Drugs, Chinese Herbal ; pharmacokinetics ; Gastrointestinal Tract ; metabolism ; Intestinal Absorption ; Male ; Polygonum ; chemistry ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the relationship between asthma and rhinitis in children.Methods One hundred and thirty children with rhinitis were divided into two groups.Among them,60 displayed rhinitis alone and 70 displayed rhinitis associated asthma.The following parameters were analyzed between two groups: age,sex,history of eczema,familial history of smoking,familial history of asthma,sensitization to allergens,level of total serum IgE,blood eosinophil count.Logistic regression analysis was used to analyze the effect of covariates on risk of rhinitis or asthma.Results History of bronchiolitis,familial history of asthma,maternal asthma and sensitization to allergens h_1(greer labs inc),d_2(dermatophagoides farinae) were significantly more frequent in asthmatic subjects.In these patients,the total serum IgE and eosinophil count were higher.Logistic regression analysis showed that a high eosinophil count and total serum IgE significantly increased the risk of developing asthma in patients with rhinitis.Conclusions In subjects with rhinitis,the occurrence of asthma is related to history of bronchiolitis,familial history of asthma,atopy,total serum IgE levels and blood eosinophilia.In rhinitis subjects,these parameters will be taken into account to manage underlying asthma.

Objective To investigate the effects of morroniside on hydrogen peroxide (H2O2)-induced calcium overload and cytotoxicity in SH-SY5Y neuroblastoma cells. Methods SH-SY5Y cells were pre-incubated with morroniside 1, 10, or 100 μmol/L for 24 h prior to exposure to H2O2 300~500 μmol/L for 18 h. The cytosolic free calcium concentration and LDH release were determined. Results The H2O2-induced cytosolic free calcium concentration decreased in the cells pre-incubated with morroniside 10 or 100 μmol/L, while the LDH release level decreased in the cells pre-incubated with morroniside 1, 10 or 100 μmol/L, comparison with the cells exposed H2O2 along. Conclusion Morroniside effects neuroprotection against H2O2-induced calcium overload and cytotoxicity in SH-SY5Y cell.