2.Expression of mieroRNA-224 in HepG2 cells and analysis of its predicted target genes
Qiong LI ; Ge WANG ; Jinlu SHAN ; Chuan CHEN ; Zhimin ZHANG ; Wen XU ; Xizhong LUO ; Dong WANG
Chinese Journal of Digestive Surgery 2008;7(4):297-299
Objective To investigate the expression of microRNA-224 in HepG2 cells and analyze its target genes to reveal its role in the carcinogenesis of hepatoma. Methods The genes with differential expression in HepG2 cells and LO2 cells were obtained by gene expression microarray analysis. The up-regulated target genes of microRNA-224 were predicted by bioinformatics method, and their functions were analyzed. Results Compared with LO2 cells, microRNA-224 was highly expressed in HepG2 cells. A total of 264 target genes of microRNA-224 were predicted, including genes involved in cell cycle, signal transduction, cell differentiation, proliferation and apoptosis. Conclusions MicroRNA-224 is highly expressed in HepG2 cells. MicroRNA-224 plays an important role in the carcinogenesis of hepatoma via regulating the expression of its target genes directly or indirectly.
3.Isolation and Purification of Exopolysaccharide from the Fermentation Broth of Bacillus sp. and Its Antioxidant Effect
Jian-Feng YUAN ; Heng CAI ; Xian-Yang SHAN ; Chuan-Xue XU ; Hong-Gui WAN ;
Microbiology 2008;0(10):-
Based on the Bacillus sp., isolated from Lop Nur Desert, the technology of separation and purification and the antioxidant effect were studied. After centrifugation and vacuum filtration, the deproteinization of supernatant was operated with Sevag reagent. The crude exopolysaccharide (EPS) was obtained by precipitation with ethanol. The optimum conditions for the isolation were as follow: pH 7.0, temperature 4?C, time 1.5 h, and material to ethanol ratio 1: 4. Dissolved in water, the crude EPS was fractional separated on activated carbon column (1.5 cm ? 24 cm), eluted with distilled water, 60% ethanol, 95% ethanol, and the main fraction was collected. Then the EPS was purified on Sephadex G-100 gel column, eluted with NaCl (0.2 mol/L). Fractions (4 mL, each) were also combined according to total sugar by phenol-sulfuric acid method and protein content was determined by Coomassie brilliant blue. The results showed that EPS was relatively homogeneous glycoprotein. The data of antioxidation in vitro showed that the EPS had a high antioxidant activity, which could quench hydroxyl radical, superoxide radical and had antilipid peroxidation activity. All of these indicated that EPS was a good natural antioxidant.
4.The effects of low frequency ultrasound on the proliferation and apoptosis of smooth muscle cells in carotid atherosclerosis
Yun-Bo CHEN ; Chang-Qing LI ; Qi SONG ; Chuan-Shan XU ; Le-Hua YU ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(12):-
Objective To investigate the effects of low frequency ultrasound (LFU) on the proliferation and apoptosis of smooth muscle cells (SMCs) in rabbits with carotid atherosclerosis(CA).Methods CA models were established in 30 New Zealand rabbits using a high fat diet and air-drying.They were randomly divided into a control group and four LFU groups:group A received 0.5 W/cm~2 LFU for 5 min/d,group B 0.5 W/cm~2 for 10 min/d, group C 1 W/cm~2,5 min/d,and group D 1 W/cm~2,10 min/d.The rabbits' carotid arteries were autopsied after 20 d of the LFU treatment.The expression of PCNA and TUNEL staining were used to explore the proliferation and apopto- sis of SMCs,and the proliferation rates (PRs) and apoptosis rates (ARs) were calculated.Results Compared with the control group,the PRs in groups B,C and D were significantly lower,while the ARs in groups B,C and D were significantly higher.There was no significant difference in ARs or PRs among groups B,C and D.Con- clusion LFU can induce SMC apoptosis and inhibit SMC proliferation in rabbits with CA.
5.HER-2 expression in advanced gastric cancer and its correlation with clinical features, outcome and prognosis.
Yan WANG ; Jian-ming XU ; Jian-zhi LIU ; Li LIN ; Fei-jiao GE ; Shan-shan LI ; Lie-jun LIU ; Chuan-hua ZHAO
Chinese Journal of Oncology 2011;33(9):671-675
OBJECTIVETo assess the HER-2 status in Chinese advanced gastric cancer patients and explore its correlation with clinical features, treatment response and prognosis.
METHODSA total of 107 patients with advanced gastric cancer treated in our hospital from December 2005 to November 2008 were included in this retrospective analysis. HER-2 status was determined by immunohistochemisty (IHC) and/or fluorescence in situ hybridization (FISH). The correlations of HER-2 status with tumor location, pathology, treatment response and prognosis were analyzed and the efficacy of different chemottherapy regimens was compared.
RESULTSThe overall positive rate of HER-2 expression was 14.7% (15/102). The HER-2 status was detected by both methods in 102 patients, and the concordance of the two methods was 66.5%. The tumor site distribution was gastroesophageal junction (GEJ) 28.0%, proximal stomach 19.4%, gastric corpus 16.1%, antrum 26.9% and whole stomach 9.7%, respectively. There was no significant difference of HER-2 status among different tumor sites (P = 0.726), and no significant correlation between HER-2 expression and differentiation (P = 0.110). Among the evaluable 51 patients treated by first-line chemotherapy, the total objective effective rate was 23.5%. The median time-to-progression was 7.47 months, and median overall survival time was 11.07 months. The effective rate was 43.8% in patients who received XP regimen chemotherapy (cisplatin + capecitabine), significantly higher than the 14.3% in patients treated with other regimens (P = 0.033). Their overall survival was 14.17 months and 9.53 months, respectively (P = 0.059). The TTP was 6.63 months in HER-2 positive patients and 7.47 months in HER-2 negative patients, with a non-significant difference (P = 0.510). However, there was a improving tendency in the efficacy and OS, showing a effective rate of 45.5% and 17.5% (P = 0.102) and OS of 14.17 months and 10.63 months, respectively (P = 0.205).
CONCLUSIONSHER-2-positivity rate in Chinese patients with advanced gastric cancer is similar to those reported in the literature. Along with the increasing use of targeted therapy and targeted agents, the efficacy and survival of gastric cancer patients is improving. HER-2-positive patients may benefit from it.
Adenocarcinoma ; drug therapy ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Capecitabine ; Cisplatin ; administration & dosage ; Deoxycytidine ; administration & dosage ; analogs & derivatives ; Disease Progression ; Esophagogastric Junction ; pathology ; Female ; Fluorouracil ; administration & dosage ; analogs & derivatives ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Receptor, ErbB-2 ; metabolism ; Retrospective Studies ; Stomach ; pathology ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology ; Survival Rate
6.Effect of Wenyun Mixture and Bran Stir-Baked Atractylodis Water Extract on Spleen-deficiency Diarrhea in Mice Based on GC-MS
Shan XU ; An KANG ; Lin-Xiu PENG ; Jin-Jun SHAN ; Shou-Chuan WANG
Journal of Nanjing University of Traditional Chinese Medicine 2018;34(2):181-184
OBJECTIVE To explore the effect of Wenyun mixture and bran stir-baked Atractylodis water extract on Spleen-deficiency Diarrhea in mice.METHODS The spleen deficiency mouse models were established by extract of Sennae Folium. Fecal short chain fatty acids(SCFAs)levels were detected and measured after treatment of different doses of Wenyun mixture and bran stir-baked Atractylodis water extract.RESULTS Compared with the model group,propionic acid level in mice feces was increased in low dose of bran fried Atractylodes water extract group significantly,propionic acid and isovaleric acid level in mice feces were increased in high dose of bran fried Atractylodes water extract group significantly,acetic acid,propionic acid and butyric acid level in mice feces were increased in high dose of Wenyun mixture group significantly.CONCLUSION We-nyun mixture and bran fried Atractylodes water extract could obviously improve short chain fatty acid level to transform spleen Qi and stop diarrhea.
7.Age-specific PSA reference ranges in Chinese men without prostate cancer.
Zhi-Yong LIU ; Ying-Hao SUN ; Chuan-Liang XU ; Xu GAO ; Luo-Man ZHANG ; Shan-Cheng REN
Asian Journal of Andrology 2009;11(1):100-103
This study is to determine age-specific prostate-specific antigen (PSA) distributions in Chinese men without prostate cancer (PC) and to recommend reference ranges for this population after comparison with other studies. From September 2003 to December 2006, 9 374 adult men aged from 18 to 96 years agreed to participate in the study. After all cases of PC were excluded, 8 422 adult men participated in statistical analysis and were divided into five age groups. Simple descriptive statistical analyses were carried out and quartiles and 95th percentiles were calculated for each age group. The age-specific PSA reference ranges are as follows: 40-49 years, 2.15 ng mL(-1); 50-59 years, 3.20 ng mL(-1); 60-69 years, 4.10 ng mL(-1); 70-79 years, 5.37 ng mL(-1). The results indicate that the ethnic differences in PSA levels are obvious. The currently adopted Oesterling's age-specific PSA reference ranges are not appropriate for Chinese men. The reference ranges of this study should be more suitable to Chinese men.
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8.Metabolomics Study of Exhaled Breath Condensate in Childhood Asthma by Gas Chromatography-Mass Spectrometry
Ming-Chen JIANG ; Shou-Chuan WANG ; Shan XU ; Qiu-Yue XU ; Jin-Jun SHAN ; Tong XIE ; Lin-Xiu PENG ; Qi-Gang DAI
Chinese Journal of Analytical Chemistry 2018;46(6):969-974
Exhaled breath condensate (EBC) was analyzed by gas chromatography-mass spectrometry (GC-MS / MS) in childhood asthma and healthy control, aiming to find the potential markers of EBC in children with asthma, and provide a scientific reference for its pathogenesis and early screening. EBC samples were collected from 21 asthmatic children (age (8. 2 ±1. 6) years) and 17 healthy children ( age (8. 1 ±1. 3) years). GC-MS / MS was used to obtain the full scan data of chemical components. Cluster analysis was performed on the two groups of metabolites by principal component analysis (PCA), and potential biomarkers were found using Metaboanalyst 3. 0 attributable metabolic pathways. The results showed that the EBC metabolic maps of asthmatic group and normal group were very different, and eight endogenous potential biomarkers were identified, suggesting that starch and sucrose metabolism, lysine degradation, aminoglycan nucleoside metabolism, phenylalanine metabolism may play important roles in the development of asthma in children.
9.Experimental study on the recycling of denatured acellular dermal matrix after burn.
Xiao-chuan WANG ; Chuan LI ; Fei SHAN ; Wen-ting WANG ; Xu-guo ZHU ; Du-yin JIANG
Chinese Journal of Burns 2012;28(3):201-206
OBJECTIVETo explore the feasibility of burn denatured acellular dermal matrix (DADM) as dermal substitute in repairing wounds.
METHODS(1) Nine Wistar rats received a deep partial-thickness scald on the back. Full-thickness wounded skin was collected on post scald day (PBD) 1, 2, and 3 (with 3 rats at each time point), and it was treated with 2.5 g/L trypsin/0.5% Triton X-100 to remove cells to prepare DADM, respectively called DADM-1 d, DADM-2 d, and DADM-3 d. Another 3 rats without scald injury were treated with the same method as above to prepare acellular dermal matrix (ADM) to serve as control. Gross and histological observations and microbiological and biomechanical tests, including ultimate tensile strength, maximum tension, stretched length at breaking, stress-strain relationship, were conducted for the resulting ADM and DADM. (2) Another 64 rats were divided into ADM group and DADM-1 d, DADM-2 d, and DADM-3 d groups according to the random number table, with 16 rats in each group. A skin flap in size of 2.0 cm×1.8 cm was raised on the back of each rat. The above-mentioned ADM, DADM-1 d, DADM-2 d, and DADM-3 d were cut into pieces in the size of 1.8 cm×1.5 cm, and they were respectively implanted under the skin flaps of rats in corresponding group. At post surgery week (PSW) 1, 3, 5, or 9, 4 rats in each group were used to observe wound healing condition and change in implants with naked eye, and histological observation of the implants was conducted. Data were processed with one-way analysis of variance and t test.
RESULTS(1) The freshly prepared DADM was milky white, soft in texture with flexibility, but poor in elasticity as compared with ADM. No epithelial structure or cellular component was observed in ADM or DADM under light microscope. Collagen fibers of DADM were seen to be thickened unevenly and arranged in disorder and eosinophilic. All microbiological results of DADM were negative. There was no statistically significant difference among DADM-1 d, DADM-2 d, and DADM-3 d in levels of ultimate tensile strength, maximum tension, stretched length at breaking, and stress-strain relationship (with F values from 0.088 to 3.591, P values all above 0.05). Values of the above-mentioned four indexes were the highest in DADM-3 d, they were respectively (13.0 ± 2.4) MPa, (61 ± 4) N, (173 ± 7)%, (45.7 ± 2.0)%. Values of the four indexes of ADM were respectively (19.0 ± 2.6) MPa, (95 ± 4) N, (201 ± 5)%, (62.5 ± 2.2)%, which were higher than those of DADM-1 d, DADM-2 d, and DADM-3 d (with t values from 6.424 to 17.125, P values all below 0.01). (2) No exudate or swelling in the wounds of rats, and no contraction or curling of implants were observed in every group at PSW 1, but inflammatory cells infiltration and Fbs inward migration were observed in the wound. At PSW 3, the growth of hair was normal in the wound in DADM-1 d, DADM-2 d, and ADM groups, but few and scattered hair grew in DADM-3 d group. The inflammatory cells decreased, while Fbs increased, and new capillaries were found to grow inwardly in each group. The decrease in inflammatory cells was slightly delayed in DADM-3 d group. At PSW 5, hair growth became normal, and implants shrank and thinned with fiber membrane wrapped densely and bundles of ingrowing large caliber blood vessels in all groups. The dermal matrix in each group merged with the surrounding normal tissue. At PSW 9, ADM and DADM became white, thin, and soft tissue sheet which was closely connected with the inner side of the flap. There was no infiltration of inflammatory cells in implants in either group. The collagen fibers arranged regularly and densely, and they were integrated with normal collagen tissue.
CONCLUSIONSThe burned DADM does not have obvious immunogenicity, but with good biocompatibility. It is prospective to become as a dermal substitute in repairing wounds.
Acellular Dermis ; Animals ; Burns ; pathology ; surgery ; Female ; Male ; Rats ; Rats, Wistar ; Reconstructive Surgical Procedures ; methods ; Skin ; injuries ; Skin Transplantation ; methods ; Skin, Artificial ; Wound Healing
10.Expression and subcellular localization of P9-ZFD protein in patients with myasthenia gravis.
Ming-shan REN ; Chuan-zhen LU ; Jian QIAO ; Hui-min REN ; Ren XU ; Ren-bao GAN
Chinese Medical Sciences Journal 2004;19(3):221-224
OBJECTIVETo express and purify the protein coded by the TRAF-type zinc finger domain of myasthenia gravis (MG)-related gene P9 (P9-ZFD) and to prepare P9-ZFD antiserum for detecting expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of patient with MG.
METHODSThe cDNA encoding P9-ZFD was amplified by RT-PCR. The cloned P9-ZFD cDNA was ligated into pET24a, and the P9-ZFD recombinant protein was induced via E. coli. BL21 (DE3) and purified by histidine affinity chromatography. P9-ZFD antiserum was prepared and its titer and specificity were determined by ELISA and Western blot. Expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of MG and control were studied.
RESULTSThe molecular weight of purified P9-ZFD protein was about 30 kD. Its purity was more than 95%. Antiserum specific for P9-ZFD was excellent. P9-ZFD protein is fully confined to the cytoplasm membrane of skeletal muscle cell of MG, obvious immunostaining was absent in the A, I, and Z bands of cytoplasm and no immunoreactivity was observed in the skeletal muscle cell of control.
CONCLUSIONP9-ZFD protein is expressed as a cytoplasm membrane-bound protein and has obvious distribution difference in the skeletal muscle cells of patient with MG and normal control.
Adult ; Cell Membrane ; metabolism ; Escherichia coli ; metabolism ; Female ; Humans ; Muscle Proteins ; biosynthesis ; genetics ; Muscle, Skeletal ; metabolism ; pathology ; Myasthenia Gravis ; metabolism ; Peptide Fragments ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Zinc Fingers