Objective:To observe the expression of microRNA (miRNA, miR) -7850 in renal cancer tissues, and to explore the effect of miR-7850 on the proliferation and migration of renal cancer cells and on the regulation of serine proteinase inhibitor B3 (SERPINB3) gene expression.Methods:Real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-7850 in renal cancer tissues and renal cancer cell lines. The renal cell carcinoma cell line with the lowest expression of miR-7850 was selected, and the negative control sequence (miR-NC) and miR-7850 mimics were transfected into renal cell carcinoma cells by Lipofectamine 2000 transfection reagent, respectively, which were defined as miR-NC group and miR-7850 group. qRT-PCR was used to detect the expression of miR-7850 in transfected renal cancer cells. The cell proliferation and migration ability after transfection were detected by cell counting kit-8 (CCK-8) method and transwell experiment. Bioinformatics prediction and dual luciferase reporter gene experiments were used to verify the target gene of miR-7850. qRT-PCR and Western blot were used to detect the expression of target genes in renal cancer cells after transfection.Results:Compared with adjacent tissues (5.95±0.44), the expression of miR-7850 in kidney cancer tissues (1.19±0.33) was lower ( P<0.01). Compared with immortalized proximal renal tubular epithelial cells (1.01±0.07), the expression of miR-7850 was lower in renal cancer cell lines ( P<0.05), and the lowest in A498 cells (0.13±0.01) ( P<0.01). The expression of miR-7850 in the miR-7850 group (7.46±0.93) was significantly higher than that in the miR-NC group (1.01±0.08) ( P<0.01), indicating successful transfection. Compared with the miR-NC group, the cell proliferation ability of the miR-7850 group was significantly reduced ( P<0.05). The number of migrating cells in miR-NC group and miR-7850 group were (139.50±12.31) and (75.09±16.05) cells, respectively, and the cell migration ability in miR-7850 group decreased significantly ( P<0.01). Bioinformatics technology shows that the target gene of miR-7850 was SERPINB3. The dual luciferase reporter gene experiment confirmed that miR-7850 can target the SERPINB3 gene ( P<0.05). Compared with the miR-NC group, the expression of SERPINB3 in cells of miR-7850 group was significantly reduced ( P<0.05), as well as the CDK4, CyclinD, Snail and Vimentin. Conclusions:miR-7850 is lowly expressed in renal cancer tissues and cell lines. miR-7850 can inhibit the proliferation and migration of renal cancer A498 cells, which may be related to its inhibition of SERPINB3 gene expression.

Objective To explore the relationship between the apoptosis of esophageal carcinoma tissue and its adjacent tissues with the survival time of patients with esophageal carcinoma by examining the apoptosis in esophageal carcinoma tissue and its adjacent tissues.Methods FCM Wag performed to detect the rates of apoptosis in 68 cases of esophageal carcinoma and their adjacent tissues and 28 cases of normal esophageal mucosae tissue.Sixty-three patients with esophageal carcinoma had been followed up and noted the survival time of every patient from the operated day to the deadline.Results The rate of apoptosis was the highest in the normal esophageal mucosae tissue(12.78±1.32)%,(7.79±1.48)% in adjacent tissue,and(4.16±2.06)% in esophageal carcinoma tissue,respectively.To follow the survival time after operation of 63 cases with esophageal carcinoma showed 60 cases in one year survival time and 35 cases in three years survival time.The rate of apoptosis in the esophageal carcinoma and its adjacent tissues wag(3.45±1.51)% and (3.96±0.94)% in the patients of one year survival time,(3.90±2.53)% and (7.89±2.27)% in the patients of three years survival time,respectively,P<0.05.Conclusions There is the phenomenon of apoptosis-escape in the esophageal carcinogenesis.There is close relationship between the apoptosis of esophageal carcinoma tissue and the survival time after operation.The apoptosis in the adjacent tissue is more important than that in the esophageal carcinoma tissue for evaluating the survival time after operation of patients with esophageal carcinoma.

Objective To study the expression of early growth response gene-1(Egr-1)in esophageal carcinoma tissue,and to explore the relationship between Egr-1 and the survival time of the patients with esophageal carcinoma.Methods RT-PCR was performed to detect the expression of Egr-1 mRNA in68 cases of esophageal carcinoma.The relationship between survival time and prognosis was analyzed.Results The expression of Egr-1 mRNA was the lowest(0.567±0.404),(0.945±0.336)and(1.201±0.347)in esophageal carcinoma tissue,para-cancerous tissue and normal esophageal mucosa tissue,respectively(F=12.709,P＜0.05,P＜0.00).21 cases showed the positive expression of Egr-1 mRNA of both the esophageal carcinoma tissue and the para-cancerous tissue.21 cases showed the positive expression of Egr-1 mRNA in a single esophageal carcinoma tissue or the para-cancerous tissue.26 cases showed the negative expression of Egr-1 mRNA both in the esophageal carcinoma tissue and the paracancerous tissue.The positive rate of Egr-1mRNA expression was 65.71%and 30.00%in the groups of the survival time for three years and the groups of the survival time for one year(P＜0.05).The survival rates in the two groups with positive expression of Egr-1 mRNA were 94.44%and 86.96%,respectively(P＞0.05).Conclusion Decreased level of EGR1 expression may be related to esophageal carcinogenesis.The expression level of Egr-1 mRNA might be associated with the survival time of the patients with esophageal carcinoma.Egr-1 expression in esophageal carcinoma tissue may be of great value for determining prognosis of esophageal carcinoma.

OBJECTIVETo investigate the estrogenic activity of icariin and genistein with estrogen-dependent human breast cancer (MCF-7) cells.

METHODMCF-7 cells were incubated with media containing 5% charcoal dextran-treated FBS in phenol red-free media for 48 h. CCK-8 kit was used to study the impact of defferent concentration of icariin and genistein on MCF-7 proliferation in vitro. Optimal concentration icariin and genistein were added into medium and total RNA was isolated after 12, 24, 36, 48 h. The gene expression of ERalpha, ERbeta, PS2, and PR were investigated by Real-time RT-PCR Total protein was also isolated and secretion of ERalpha, ERbeta, PS2, and PR were examined by Western blot.

RESULT10 micromol x L(-1) icariin and genistein could promote the proliferation of MCF-7 evidently. However, the ability of genistein to promote the proliferation was better than icariin. With the concentration of 10 micromol x L(-1), genistein group had a stronger expression of ERa, PS2 and PR mRNA levels than icariin while ERbetaexpression had no significant difference in two group. The same effects were detected by western blotting.

CONCLUSIONBoth genistein and icariin have a strong estrogen-like effect, but the estrogenic activity of genistein is stronger than icariin. It showed that the activity of icariin is stron-ger than genistein to promote ROB maturation. So it must be that icariin promotes the maturation of osteoblasts in vitro by a estogen-independent mechanism.

Cell Proliferation ; drug effects ; Estrogen Receptor alpha ; genetics ; metabolism ; Estrogen Receptor beta ; genetics ; metabolism ; Estrogens ; pharmacology ; Flavonoids ; pharmacology ; Gene Expression Regulation ; drug effects ; Genistein ; pharmacology ; Humans ; MCF-7 Cells ; Osteoblasts ; cytology ; drug effects ; metabolism ; Presenilin-2 ; metabolism

Objective:To explore the expression of long non-coding RNA LINC00630 in bladder cancer cell lines, and to explore the effect of interference with its expression in vitro on the proliferation and migration of bladder cancer cells.Methods:Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of LINC00630 in bladder cancer cell lines 5637, BIU-87, T24, J82 and normal bladder epithelial cell line SV-HUC-1. The bladder cancer cell line with the highest LINC00630 expression was selected for follow-up experiments, then the cell line infected with the control lentivirus was used as the control group, and the cell line infected with the lentivirus that could interfere with the expression of LINC00630 was used as the experimental group. qRT-PCR was used to detect the expression of LINC00630 in the two groups of cells. MTS method and cell scratch test were used to detect the proliferation and migration abilities of cells in the two groups. qRT-PCR was used to detect the expression of neuregulin 1 (NRG1) mRNA in the two groups of cells, and Western blot was used to detect the expressions of NRG1 protein, cell proliferation-related proteins (cyclin D3 and CDK2) and cell migration-related proteins (Vimentin and N-cadherin) in the two groups of cells.Results:Compared with SV-HUC-1 cells (1.05±0.17), the expression of LINC00630 was significantly increased in all bladder cancer cell lines (all P < 0.01), and the expression was highest in J82 cells (relative expression 5.83±0.42). Compared with J82 cells of the control group, the expression of LINC00630 in J82 cells of the experimental group decreased (0.18±0.02 vs. 1.00±0.05, t=14.36, P < 0.01); from day 2 of transfection, the cell proliferation activity of the experimental group was lower than that of the control group (all P < 0.05). The cell scratch closure rate of the experimental group was lower than that of the control group [(27.4±7.1)% vs. (66.0±5.4)%, t = 4.31, P < 0.01]. Therelative expression of NRG1 mRNA in the experimental group was lower than that in the control group (0.34±0.03 vs. 1.07±0.24, t = 2.99, P < 0.05). Compared with the control group, the expressions of NRG1 protein, cell proliferation-related proteins and cell migration-related proteins in the experimental group were reduced. Conclusions:LINC00630 is up-regulated in bladder cancer cell lines, and interference with LINC00630 may inhibit the proliferation and migration of J82 cells by down-regulating the expression of NRG1 gene. LINC00630 may be a new molecular target for the treatment of bladder cancer.

ObjectiveTo evaluate the intelligence profile,personality and behaviour disorders of children with primary generalized tonic and/or clonic seizures.MethodsUsing Chinese modification of Wechsler Intelligence Scale for Children,Eysenk Personality Questionnaire(EPQ),and Achenbach Child Behaviour Check List,neuropsychological assessment of 35 cases aged 6-14 years and the normals with the similar age and sex were makenand compared.ResultsExcept picture completion ,all the subtests scales of children with epilepsy were significantly lower than those of the controls (P<0.01). A significant difference of average FIQ(P<0.001) was observed between experimental group (85.86) and control group (103.60).There were 13 cases(37.14%) in experimental group whose FIQ were in broderline range, FIQ of 5 cases(14.29%) was in lower range,while none abnormal in the controls.The children with epilepsy had more abnormal behaviours than that of the controls(abnormal rates were 38.71% and 6.45% respectively).Statistical analysis showed no difference of EPQ scales between the two groups.Conclusions Children with primary generalized epilepsy maybe complicate with cognitive deficit and behaviour disorder.

OBJECTIVETo establish osteoblast model, primary cilla model was removed by chloral hyrate, observe effects of osteoblast primary cilla moved on enhancing ALP staining and calcified nodules staining in electromagnetic field.

METHODSThree 3-day-old male SD rats weighed between 6 and 9 g were killed, cranial osteoblast was drawed and adherencing cultured respectively. Cells were subcultured and randomly divided into 4 groups until reach to fusion states. The four groups included chloral hydrate non-involved group (control group), 2 mM, 4 mM and 8 mM chloral hydrate group, and cultured in 37 °C, 5% CO2 incubator for 72 h. Morphology of primary cilla was observed by laser confocal scanning microscope, and incidence of osteoblast primary cilia was analyzed by Image-Pro Plus 6.0 software. Cells in the correct concentration group which can removed cillia most effectively were selected and divided into 3 groups, including control group (C), Electromagnetic fields group (EMFs), and EMFs with 4 mM chloral hydrate group. DMEM nutrient solution contained 10%FBS were added into three groups and cultured for 9 days and formation of ALP were observed by histochemical staining of alkaline phosphatase. After 12 days' cultivation, formation of mineralization nodes was observed by alizarin red staining.

RESULTSCompared with control group and 2mM chloral hydrate group,4 mM chloral hydrate group could effectively remove osteoblast primary cilla (P<0.01). Removal of osteoblast primary cilla could weaken the formation of ALP and mineralization nodes in osteoblast in EMFS. Compared with EMFs group, the area of ALP and mineralization nodes in EMFs with 4 mM chloral hydrate group were decreased obviously (P<0.01).

CONCLUSION4mM chloral hydrate could effectively remove osteoblast primary cilia. Primary cilla participate in EMFs promoting formation of ALP and mineralization nodes in osteoblast and provide new ideas for exploring mechanism of EMFs promoting osteoblast maturation and mineralization.

Alkaline Phosphatase ; metabolism ; Animals ; Cell Culture Techniques ; instrumentation ; methods ; Cells, Cultured ; Chloral Hydrate ; pharmacology ; Cilia ; drug effects ; enzymology ; physiology ; Male ; Osteoblasts ; cytology ; enzymology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETuberculosis remains a severe public health issue, and the Beijing family of mycobacterium tuberculosis (M. tuberculosis) is widespread in East Asia, especially in some areas in China, like Beijing and Tianjin. This study aimed at determining the mutation patterns of drug-resistant Beijing strains of M. tuberculosis isolated from Tianjin, China.

METHODSA total of 822 M. tuberculosis isolates were screened for drug resistance by an absolute concentration method and the genotype was identified by PCR. 169 drug-resistant isolates of the Beijing family were analyzed for the potential mutations in the rpoB, katG, inhA promoter region and in rpsL, rrs and embB genes, which are associated with resistance to rifampin (RFP), isoniazid (INH), streptomycin (SM) and ethambutol (EMB) respectively by PCR and DNA sequencing.

RESULTSFifty-eight out of 63 RFP-resistant isolates were found to carry the mutations within the 81-bp RFP resistance determining region (RRDR) of the rpoB gene and the most frequent mutations occurred at codon 531 (44.4%), 526 (28.6%), and 516 (7.9%) respectively. 16 mutation patterns affecting 12 different codons around the RRDR of rpoB were found. Of 116 INH-resistant isolates, 56 (48.3%) had the mutation of katG 315 (AGC-->ACC) (Ser-->Thr), 3 (2.6%) carried S315N (AGC-->AAC) and 27 (16.0%) had the mutation of inhA-15A-->T. 84 out of 122 SM-resistant isolates (68.9%) displayed mutations at the codons 43 or 88 with AAG-->AGG (Lys-->Arg) of the rpsL gene and 22 (18.0%) with the mutations at positions 513A-->C, 516C-->T or 905 A-->G in the rrs gene. Of 34 EMB-resistant isolates, 6 had mutation with M306V (ATG-->GTG), 3 with M306I (ATG-->ATT), 1 with M306I (ATG-->ATA), 1 with D328Y (GAT-->TAT), 1 with V348L (GTC-->CTC), and 1 with G406S (GGC-->AGC) in the embB gene.

CONCLUSIONThese novel findings extended our understanding of resistance-related mutations in the Beijing strains of M. tuberculosis and may provide a scientific basis for development of new strategies for diagnosis and control of tuberculosis in China and other countries where Beijing strains are prevalent.

Base Sequence ; China ; DNA Primers ; Drug Resistance, Microbial ; Mycobacterium tuberculosis ; genetics ; Polymerase Chain Reaction

Objective To observe the influence of long-term treatment with inhaled corticosteroid on biochemical bone indexes and bone mineral density (BMD) in children with asthma. Methods The design was a randomized, paralleled group study with 3 low dose regiments of 100, 200,300 micrograms of budesonide per day in 45 children with asthma aged 5-8 years old for 12 months. Before inhaled corticosteroid therapy and 6th,12th month,clinical effects were observed and lung function(FEV1) was measured; concentration of serum osteocalcin(OST),insulin-like growth factor-1(IGF-1),bony alkaline phosphatase (BALP) and urinary deoxypyridinolin: creatinine (DPD/Cr) were measured; BMD was examined by dual energy X-ray absorptiometry. Results Clinical evaluation was improved and there was significant increase in FEV1 of asthmatic children. The amount of serum OST was slightly higher,yet no significant compared with that of normal control group. There was significant increase of serum BALP in asthmatic children after treatment; there was significant increase in serum IGF-1 of patients group after treatment compared with in normal children at the same age group; there was significant decrease in urinary DPD/Cr after treatment.There was no significant decrease in BMD before and after treatment at the hip (neck of femur , trochanter of femur ,Ward′s triangle),the lumber area of the spine (L2-4) and forearm (ultradis, distal). Conclusion Long-term treatment with low does corticosteroid dose not restrictedly affect bone metabolism and BMD in children with asthma.

AIMA series of new 1,4-pentadien-3-one derivatives were synthesized to search for new Eight novel hydroxylated non-steroidal anti-inflammatory drugs (NSAIDs) with potent activity.

METHODSE,E-1-(3'-indolyl)-5-( substituted phenyl)-1,4-pentadien-3-one derivatives were synthesized by means of aldol condensation and characterized by 1H NMR, ESI-MS and element analysis. Their anti-inflammatory activity in vitro were evaluated.

RESULTSPreliminary in vitro pharmacological tests showed that all compounds exhibited anti-inflammatory activity.

CONCLUSIONCompounds 4d and 4e exhibited potent anti-inflammatory activity and their anti-inflammatory activity was comparable to resveratrol, and were worthy of further study.

Alkadienes ; chemical synthesis ; pharmacology ; Animals ; Anti-Inflammatory Agents ; chemical synthesis ; pharmacology ; Indoles ; chemical synthesis ; pharmacology ; Macrophages, Peritoneal ; cytology ; metabolism ; Male ; Mice ; Tumor Necrosis Factor-alpha ; secretion