10.3969/j.issn.2095-4344.2013.25.021

Objective To investigate the effects of cyclooxygenase-2 (COX-2) selective inhibitor combined with 5-lipoxygenase (5-LOX) inhibitor docebenone (AA861) on cell proliferation and migration of human colon cancer cell line HT-29. Methods Cultured in vitro of HT-29 cells in high metastatic colon cancer, A A861 and celecoxib on colon cancer cell drugs for 48 h, the cell proliferation was assayed by MTT; the migration of cell was detected by Transwell; immunofluorescence staining of intracellular changes in ICAM-1 protein, RT-PCR detect the gene expression of ICAM-1 and VEGF. Results MTT and Transwell experiments showed that the inhibition on celecoxib and AA861 on colon cancer was dose-dependent and time-dependent. And the inhibition of AA861 and celecoxib 100 μmol/L alone on colon cancer cells proliferation were 43.2 % and 42.8%, and when the two drugs 100 μmol/L combined on colon cancer cells the inhibition was 53.8%, and the difference between alone group and combined group was statistical (P ＜0.001). The inhibition of AA861 and celecoxib 100 μmol/L alone on colon cancer cells migration were 32.0 % and 29.3%, and when the two drugs 100 μ,mol/L combined on colon cancer cells the inhibition is 57.8 %, and the difference of between alone group and combined group was statistical (P ＜0.001). Immunefluorescence staining and RT-PCR results suggested that the two drugs can inhibit ICAM-1 and VEGF protein and gene expression, and when the two drugs combined, a stronger inhibition effect appeared than used alone (P＜0.001). Conclusion Low-dose celecoxib and AA861 combined has a synergistic inhibited effect on colon cancer cells invasion and metastasis, and the mechanism relates with the VEGF and ICAM-1 expression.

Objective To study the effect of 5-lipoxygenase(5-LOX) inhibitor nordihyroguaiaretic acid (NDGA) combined the selective cyclooxygenase-2 (COX-2) inhibitor Celecoxib on the apoptosis of human colon carcinoma cell line HT-29. Methods Different concentration of NDGA and Celecoxib combinations were used to process cancer cell, and thiazolyl blue tetrazlium bromide (MTT) and phase contrast microscope and Annexin V/PI fluorescence staining and reverse transcription polymerase chain reaction (RT-PCR) were used to study the proliferation inhibited effect and apoptosis induced effect caused by combination of NDGA combined Celecoxib. Results MTT results showed that the viability of NDGA group, Celecoxib group and the group of NDGA combined Celecoxib (0.432±0.024,0.425±0.013,0.303±0.014 vs 0.693±0.018,t＝18.79,25.75,37.64,P＜0.01) was obviously lower than control group. The group of NDGA combined Celecoxib was significantly lower than NDGA group or Celecoxib group (t＝10.21, 14.14,P＜0.01). Under inverted phase contrast microscope, cell morphology significantly changed, and the group of NDGA combined Celecoxib changed most obviously. Apoptosis was observed by laser scanning confocal microscope (LSM) after NDGA and Celecoxib were used to process the HT-29. RT-PCR showed that up-regulation of Caspase-3 after treatment, and the combination of two drugs increased the most. Conclusions NDGA combined Celecoxib inhibited proliferation and induced apoptosis in human colon carcinoma cell line HT-29, and combined therapy had better effect than that of any drug used separate-ly. The mechanism may be associated with up-regulation of Caspase-3.

Objective:To describe the status of subjective well-being among junior high school students in a city of Jiangxi Province and to explore the relationship between family functioning and subjective well-being.To explore the relationship between growth environment and subjective well-being among ju-nior high school students in the perspective of family functioning and to provide basis and data for the mental health education among junior high school students.Methods:630 students in a junior high school in Jiangxi Province were sampled by stratified cluster sampling,and 103 junior high school students from 18 villages were sampled by household survey.They were investigated by subjective well-being scale for adolescents and Family Assessment Device.Results:(1)The overall subjective well-being of junior high school students was in the lower-middle level.Regarding all the dimensions of subjective well-being,they had the highest satisfaction on family and friendship,lowest academic satisfaction,low positive as well as passive emotion.(2)Girls had higher satisfaction on friendship and freedom than that of boys;children from families with more than one kids had more negative emotion than that of single child;boarding students had higher satisfaction on family,academics and freedom than that of non-boarding students;family residence and structure affected the satisfaction on family;grade and learning achievement affected some dimensions of subjective well-being among junior high school students.(3)Pearson correlation analysis and multiple linear regression analysis showed that some dimensions of family functioning were related to certain dimensions of subjective well-being among junior school students,and family functioning could predict the level of subjective well-being.Conclusion:The overall subjective well-being of junior high school students is in the lower-middle level,and further improving is needed.Family functioning has a great influence on junior school students' subjective well-being.Junior high school students who have healthy family functioning are more likely to have higher life satisfaction,and tend to experience higher subjective well-being.

Objective To observe the effects of stromal cell-derived-factor-1(SDF-1) on the function of endotheli?al progenitor cells(EPCs)of peripheral blood in patients with diabetes, and to discuss the effects of PI3K/AKT signaling path?way on the role of SDF-1 in EPCs. Methods The peripheral blood samples (30 mL) were collected in 10 diabetes patients (DM group) and 10 healthy controls (HC group). (1) The 100μg/L SDF-1 was added in intervention group. EGM-2MV was added in non-intervention group. The Boyden chamber and in vitro angiogenesis kit were used to analyze the migration and in vitro angiogenesis of EPCs. (2) Cultured EPCs were divided into blank control group, 1μg/L SDF-1 group, 10μg/L SDF-1 group, 100μg/L SDF-1 group, pure AMD3100 group and 100μg/L SDF-1+AMD3100 group. AKT protein expression lev?els of endothelial progenitor cells were detected by Western blot assay in each group. Results (1) Without intervention with SDF-1, EPCs’migration and angiogenesis ability were lower in DM group than those in HC group. After intervention with SDF-1, the migration and angiogenesis ability were enhanced in two groups, but the increased level was higher in DM group than that of HC group. (2) Under the same concentration, AKT protein expression level was significantly lower in DM group than that in HC group (P<0.01). AKT protein expression levels were increased with the increased levels of SDF-1 in DM group and HC group (P<0.05). AKT protein expression was significantly lower in 100μg/L SDF-1+AMD3100 group than that of 100μg/L SDF-1 group (P<0.05). Conclusion SDF-1 can increase the chemotactic migration and angiogenesis ability of EPCs in peripheral blood, especially for patients with diabetes. The effects of SDF-1 on EPCs were related to the PI3K/AKT signaling pathway.

Objective To compare the effects of electroacupuncture at points Shangjuxu(ST37), Zusanli(ST36), Xiajuxu(ST39) and Yanglingquan(GB34) on colonic expressions of interleukin-1b (IL-1b) and nicotinic acetylcholine receptor a7 mRNA (nAchRa7mRNA) in ulcerative colitis rats and investigate if large intestine lower He-Sea point Shangjuxu has relative specificity to fu organ diseases. Method Seventy healthy SD rats were randomized into blank, model, Shangjuxu, Zusanli, Xiajuxu, Yanglingquan and Chengjin groups, 10 rats, half male and half female, each. A rat model of ulcerative colitis was made by induction of 2-4-6 three nitrobenzene sulfonic acid/ethanol solution enema in every group except the blank group. After successful model making and ten days of treatment, rat colonic mucosal ulcers and inflammation were observed macroscopically, colonic IL-1bcontent was measured by ELISA and the expression of nAchRa7mRNA was determined by RT-PCR. Result Compared with the model group, colonic lesions were reduced in varying degrees, colonic IL-1b content was significantly lower and the expression of nAchRa7mRNA was higher in every acupoint group (P<0.05, P<0.01);the colonic ulcer score was lower in the Shangjuxu and Zusanli groups (P<0.05, P<0.01). Compared with the Shangjuxu group, colonic expression of nAchRa7mRNA was lower in the other four acupoint groups (P<0.01); colonic mucosal ulcers and inflammatory lesions were more severe and the colonic ulcer score and the IL-1bcontent were higher in the Xiajuxu, Yanglingquan and Chengjin groups (P<0.05, P<0.01). Conclusion The mechanism of electroacupuncture treatment for ulcerative colitis may be that it regulates abnormal immunologic function by modulating IL-1b and nAchRa7mRNA and reduces mucosal lesions. The overall therapeutic effect of Shangjuxu is better than those of Zusanli, Xiajuxu, Yanglingquan and Chengjin, indicating that Shangjuxu has relative specificity to fu organ large intestine.

Objective To explore the effect of gravitational traction on height change of degenerative disc of New Zealand white rabbit.Methods In order to create intervertebral disc degeneration models, the right front side of the annulus fibrosus of lumbar vertebrae of rabbits were damaged by 16-gauge needle. After confirming the success of modeling, 20 animal models were randomized into 2 groups, 15 of which received a routine gravitational traction using a model of our own design, 30 min per day, for 8 weeks, while the control group received no treatment at all. Radiographic exam were performed in the 2nd, 4th, 6th and 8th week, after intervention and the disc height index percent (DHI%). Results Compared to the baseline, DHI% increased significantly 2 and 4 weeks after gravitational traction (P<0.05), and there was no further effect as the traction continued. However, compared to the control group, an improvement of DHI% was found on all the time points in the gravitational traction group (P<0.001). Conclusion Gravitational traction may delay and inhibit intervertebral disc degeneration by increasing height of degenerated disc of New Zealand white rabbit.

OBJECTIVETo determine the association between the polymorphism of angiotensin converting enzyme (ACE) gene and Alzheimer's disease (AD).

METHODSThis case-control study involved 201 AD patients and 257 healthy subjects matched for age and gender as the control group. Polymerase chain reaction amplification and matrix-assisted laser desorption/ ionization time of flight mass spectrometry were used to examine the rs4291, rs4309, and rs4343 of ACE gene, and the difference in genotypes, allelotype frequencies and haplotype frequencies were analyzed between the two groups.

RESULTSNo statistic difference was found in the genotype and allelotype frequencies of rs4291 locus between AD and control groups (P>0.05). A significant difference was found in the genotype and allelotype frequencies of rs4309 between the two groups with a significant increase in the C allelotype frequency in AD group (OR=1.917, 95% CI=1.431-2.568, P<0.05). The difference in the genotype frequency of rs4343 was not significant between the two groups, but the allelotype frequencies differed significantly with a decreased A allelotype frequency in AD group(OR=0.714, 95% CI=0.532-0.957, P=0.024). Analysis of the linkage disequilibrium among the loci of rs4291, rs4309 and rs4343 showed a D' all above 0.65 between one another. Haplotype analysis confirmed the existence of 5 haplotypes, namely ATA, ACA, TCA, TCG and TTG, indicating a negative correlation between haplotype ATA and AD occurrence (OR=0.558, 95% CI=0.420-0.741, P<0.05) and positive correlations of haplotype ACA and TCA with AD occurrence (ACA: OR=4.883, 95% CI=2.267-10.518, P<0.05; TCA: OR=2.269, 95% CI=1.083-4.754, P<0.05).

CONCLUSIONThe polymorphism of rs4291 may have no relation with the incidence of AD. Polymorphisms of s4309 and rs4343 may be related to AD, and ATA, ACA and TCA haplotypes composed of rs4291/rs4309/rs4343 may be related to AD.

Alzheimer Disease ; genetics ; Case-Control Studies ; Gene Frequency ; Genotype ; Haplotypes ; Humans ; Linkage Disequilibrium ; Peptidyl-Dipeptidase A ; genetics ; Polymorphism, Single Nucleotide

OBJECTIVEOsseointegration of orthodontic microscrew implant is influenced by tooth extraction. This study aims to evaluate the safety margin of the osseointegration of orthodontic implants by investigating the healing process of the implant-bone interface through histopathological studies and quantitative determination.

METHODSTwelve male beagles were selected and randomly divided into four groups. An orthodontic microscrew was implanted beside the tooth extraction area. Animals were killed in 1, 3, 8, and, 12 weeks to investigate tissue response. Histomorphological observation and bone implant contact ratio (BIC) tests were performed at different healing time after implantation.

RESULTSA new bone was formed on the implant-bone interface of the control group. Bone resorptions were also detected in the experimental group 3 weeks after implantation. The BIC level of the control groups increased during the first 8 weeks; no change was observed anymore after the 8th week. On the other hand, the BIC value in the experimental group decreased in the first 3 weeks. It then increased rapidly and reached its peak of 80.08% in the 8th week. No significant difference wa s found between the experimental and control groups in the first 3 weeks. After the 3rd week, the BIC value of the experimental group (44.35%) was lower than that of the control group (55.46%) (P < 0.01).

CONCLUSIONThe healing process after implantation was influenced by tooth extraction. Bone resorption was detected at an early stage. However, vigorous bone remodeling was observed subsequently.

Animals ; Bone Remodeling ; Bone and Bones ; Dental Implantation, Endosseous ; Dental Implants ; Dogs ; Male ; Mandible ; Osseointegration ; Prostheses and Implants ; Tooth Extraction ; Wound Healing

Objective:To explore the expressions of long non-coding RNA LINC00673 and ISG15 protein in pancreatic cancer and their clinical significances.Methods:The clinical data of 57 patients diagnosed as pancreatic ductal carcinoma (PDAC) at the Affiliated Cancer Hospital of Xiangya Medical College of Central South University from January 2014 to December 2018 were retrospectively analyzed. The relative expressions of LINC00673 in pancreatic cancer tissues and paracancerous normal tissues (within 3 cm from the edge of cancer tissues) were examined by using quantificational reverse transcription-polymerase chain reaction (qRT-PCR). The ISG15 protein expressions in pancreatic cancer tissues and paracancerous normal tissues were examined by using immunohistochemistry. The difference in LINC00673 expression between ISG15 protein positive and negative patients was compared. The correlation between LINC00673 and ISG15 protein expressions in pancreatic cancer was analyzed by Spearman rank correlation analysis. Moreover, the correlations of LINC00673 and ISG15 protein expressions with clinical stage and pathological classification of pancreatic cancer patients were analyzed.Results:The positive expression of ISG15 protein in pancreatic cancer tissues was 40.4% (23/57), which was higher than that in paracancerous normal tissues [15.8% (9/57)] ( χ2 = 7.90, P = 0.004), and the relative expression of LINC00673 in pancreatic cancer tissues was 0.99±0.36, which was lower than that in paracancerous normal tissues (1.26±0.41) ( t = 4.80, P < 0.001). For 23 (40.4%) ISG15-positive patients and 34 (59.7%) ISG15-negative patients, the relative expression of LINC00673 was 0.77±0.46 and 0.45±0.27 ( P < 0.001). Spearman analysis showed that there was a correlation between LINC00673 and ISG15 protein expressions ( ρ = -0.429, P = 0.001). The relative expression of LINC00673 decreased in patients with low differentiated or undifferentiated tumor, vascular invasion and lymph node metastasis (all P < 0.05), but there was no correlation between LINC00673 expression and patients' age, tumor site, preoperative CA199 level, and TNM stage (all P > 0.05); ISG15 protein expression increased in patients with low differentiated or undifferentiated tumor, TNM stage Ⅲ-Ⅳ, vascular invasion and lymph node metastasis (all P < 0.05), but there was no correlation between ISG15 protein expression and patients' gender, age, tumor site, and preoperative CA199 level (all P > 0.05). Conclusions:The expression of LINC00673 in pancreatic cancer is related to vascular invasion, tumor differentiation degree and lymph node metastasis, and the expression of ISG15 in pancreatic cancer is related to vascular invasion, tumor differentiation degree, lymph node metastasis and TNM stage. The combined detection of LINC00673 and ISG15 protein could be a valuable prognostic indicator for pancreatic cancer. The therapies targeting LINC00673 and ISG15 protein signaling pathways are expected to be a potential option for immunotherapy of pancreatic cancer.