1.Changes of large-conductance calcium-activated potassium channel in gastric smooth muscle cells of diabetic gastroparesis rats
Jianhui XU ; Xiaoyu HOU ; Hongxian ZHAO
Chinese Journal of Pathophysiology 2015;(7):1288-1293
[ ABSTRACT] AIM: To discuss the relevance between the pathogenesis of diabetic gastroparesis and the large-conductance calcium-activated potassium channels ( BKCa ) in gastric smooth muscle cells.METHODS:The SD rats were randomly divided into control group and model group.The gastric smooth muscle cells of the SD rats were enzymatically iso-lated in a low calcium solution containing papain.The current was recorded by patch clamp single channel recording tech-nique.The expression of KCNMA and KCNMB1 were observed by the method of immunohistochemistry.RESULTS:The value of BKCa single channel conductance was (220.10 ±10.90) pS;the channels had distinct voltage dependent and cal-cium dependent characteristics.In outside-out patch (Vm =+30 mV), the activation of BKCa was blocked by 200 nmol/L IbTX completely.Compared with control group, the open probability and amplitude of current in model group significant-ly increased, while the mean open time and mean close time significantly decreased.Compared with control group, the ex-pression of KCNMB1 in model group was significantly increased.CONCLUSION: Up-regulation of β1-subunit and in-crease in BKCa functional activities may be associated with diabetes gastroparesis in rats.
2.Development of microsatellites of Dendrobium officinale and its application in purity identification of germplasm
Minglu XIE ; Beiwei HOU ; Li HAN ; Yanhong MA ; Xiaoyu DING
Acta Pharmaceutica Sinica 2010;45(5):667-72
Simple sequence repeat (SSR) was used to investigate the genetic diversity and structure of Dendrobium officinale. A total of 15 primer pairs with stable and repeatable polymorphism were screened out from 60 SSR primer pairs developed by the method of microsatellite enrichment by magnetic beads. Forty-eight samples of Dendrobium officinale were analyzed in genetic polymorphism. These loci were polymorphic and displayed 3 to 9 alleles per locus with a mean number of 6.1. The observed and expected heterozygosities ranged from 0.60 to 0.85 and from 0.49 to 0.85 respectively. The polymorphic information content (PIC) of each SSR locus varied from 0.437 to 0.829 with an average of 0.702. Fifteen primer pairs were used in Dendrobium cross-species amplification and totally 13 primer pairs were proved to have the transferability in D. officinale related species. In addition, 500 tissue culture plantlets of D. officinale were tested for purity identification by means of PCR amplification with four SSR primer pairs. The results showed that SSR technique is a feasible, simple and inexpensive method for determining adulterants in germplasm identification.
3.Expression of DNA methyltransferases in liver cancer and its clinical significance
Jianxin PENG ; Xiaoyu YIN ; Xun HOU ; Jianen WANG ; Lijian LIANG
Chinese Journal of Digestive Surgery 2011;10(2):124-128
Objective To investigate the expression of DNA methyltransferases (DNMTs) in liver cancer and its clinical significance. Methods The specimens of liver cancer tissues, adjacent tissues, cirrhotic tissues and chronic hepatitis tissues were collected from 50 patients who received radical resection at the First Affiliated Hospital of Sun Yat-Sen University from July 2007 to April 2008. The mRNA and protein expressions of DNMT1,DNMT3a and DNMT3b in liver cancer tissues, adjacent tissues, cirrhotic tissues and chronic hepatitis tissues were detected by real-time quantitative PCR and immunohistochemical staining. The mRNA expression of DNMTs in the liver cancer tissues was compared with those in the adjacent tissues, cirrhotic tissues and chronic hepatitis tissues by using t test and Mann-Whitney U test. The correlation between the protein expression of DNMTs in the liver cancer tissue and the clinicopathological features was analyzed by chi-square test or Fisher exact test, and the tumor-free survival time was analyzed by using Kaplan-Meier method and the difference in tumor-free survival rate between different patients was analyzed by Log-rank test. Results The mRNA expressions of DNMT1, DNMT3a and DNMT3b in the liver cancer tissue were 2.57, 2.29 and 4.86 times higher than those in the adjacent tissues (t = 3.94, 2. 72, 4. 06, P < 0.05 ). The mRNA expressions of DNMT1, DNMT3a and DNMT3b were 2.38,2.14 and 4.66 times higher than those in the cirrhotic tissues, and 6.12, 4.58 and 12.99 times higher than those in the chronic hepatitis tissues. The mRNA expressions of DNMT1, DNMT3a and DNMT3b in the liver cancer tissue were significantly higher than those in the cirrhotic tissues and chronic hepatitis tissues ( U = 587.5,730. 0,562.5; 65.5, 64.5, 71.0, P < 0.05). The protein expression of DNMT1 was correlated with the size, number,TNM stages and vascular invasion of tumors ( x2 = 4.08, 5.95, 4.08, P < 0.05 ). The protein expression of DNMT3a was correlated with the size, number and TNM stages of tumors (x2 = 4.08, 5.95, 4.08, P < 0.05 ).The mean tumor recurrence time of patients with low expressions of DNMT1 and DNMT3a were 9.4 and 8.7 months, which were significantly longer than 5.0 and 3.2 months of those with high expressions of DNMT1 and DNMT3a (x2 =3.89, 9.91, P<0.05). Conclusions DNMTs play an important role in hepatocarcinogenesis.High expressions of DNMT1 and DNMT3a are correlated with the postoperative recurrence of liver cancer, which are valuable prognostic factors for liver cancer.
4.Sequence analysis of LEAFY homologous gene from Dendrobium moniliforme and application for identification of medicinal Dendrobium.
Wenrui XING ; Beiwei HOU ; Jingjiao GUAN ; Jing LUO ; Xiaoyu DING
Acta Pharmaceutica Sinica 2013;48(4):597-603
The LEAFY (LFY) homologous gene of Dendrobium moniliforme (L.) Sw. was cloned by new primers which were designed based on the conservative region of known sequences of orchid LEAFY gene. Partial LFY homologous gene was cloned by common PCR, then we got the complete LFY homologous gene Den LFY by Tail-PCR. The complete sequence of DenLFY gene was 3 575 bp which contained three exons and two introns. Using BLAST method, comparison analysis among the exon of LFY homologous gene indicted that the DenLFY gene had high identity with orchids LFY homologous, including the related fragment of PhalLFY (84%) in Phalaenopsis hybrid cultivar, LFY homologous gene in Oncidium (90%) and in other orchid (over 80%). Using MP analysis, Dendrobium is found to be the sister to Oncidium and Phalaenopsis. Homologous analysis demonstrated that the C-terminal amino acids were highly conserved. When the exons and introns were separately considered, exons and the sequence of amino acid were good markers for the function research of DenLFY gene. The second intron can be used in authentication research of Dendrobium based on the length polymorphism between Dendrobium moniliforme and Dendrobium officinale.
5.Evaluation of sixteen personality factor questionnaire to establish Personality Characteristics Questionnaire and post personality model of nurses
Lijuan YANG ; Jin LIU ; Xiaohong HOU ; Xiaoyu GUO
Chinese Journal of Practical Nursing 2016;32(17):1289-1293
Objective To understand the personality characteristics of excellent and generalnurses, extract personality characteristics from excellent nurses, and to establish Personality Characteristics Questionnaire and post personality model of nurses, and explore influencing factors of nurses personality characteristicsfor providing a theoretical reference in selection and training of nurses. Methods A total of 523 staff nurses from one class 3-A hospital general hospitalsin Shandong province were investigated with Sixteen Personality Factor Questionnaire(16PF). Results Excellent nurses possessed higher dominance, rule-consciousness, social boldness, shrewdness, perfectionism and lower apprehension than general nurses (t=-2.54-2.92, P<0.05), then six personality characteristics were extracted from excellent nurses to establish Personality Characteristics Questionnaire and post personality model of nurses; The difference was statistically significant in perfectionism with different age, work experience, and professional title in excellent of nurses (F=4.04-7.36, P < 0.05). Conclusions 16PF is suitable for evaluating personality characteristics of nurses, the personality characteristics questionnaire and post personality model can be used as selection criteria for new nurses.
6.A comparative study of nurse work environment between military hospital and civilian hospital
Ping LIANG ; Yuan WANG ; Ming HOU ; Xiaoyu YANG
Chinese Journal of Practical Nursing 2013;29(21):7-9
Objective To investigate the current situation and the differences of nurse work environment between a military hospital and a civilian hospital.Methods By quota sampling,a questionnaire survey was conducted among a total of 1244 nurses in a military hospital and a civilian hospital in Urumqi.Results The nurse work environment in both hospitals was favorable overall.The total scores of nursing environment and subscale scores of the military hospital were lower than those of the civilian hospital.There were no statistical differences between the two hospitals on their perceptions of the collaboration with physicians as well as leadership and ability of nurse manager.Conclusions The nurse work environ-ment in the civilian hospital was a little more favorable than the military one.Both of their managers,espe-cially the military hospital,should pay more attention to manpower and resources as well as nurses'involve-ment in hospital affairs in order to further optimize nurse work environment.
7.Clinical application and value of dual-source spiral CT enhanced scan and multiplanar reformatting in diagnosis of gastric cancer
Jing PENG ; Qide YAN ; Jiafei CHEN ; Xiaoyu HOU
Journal of Regional Anatomy and Operative Surgery 2015;(2):185-186,187
Objective To discuss the clinical application and value of dual-source spiral CT enhanced scan and multiplanar reformatting ( MPR) in the diagnosis of gastric cancer. Methods The 335 patients with gastric cancer who conducted dual-source spiral CT enhanced scan of upper abdomen under the hypotonic state before the definite pathological diagnosis were retrospectively analyzed,and had multi-di-mensional multiplanar reconstruction for the enhanced thin-section CT data by the multiplanar reformatting technique. Results Of 335 ca-ses,288 cases were detected by simple axial data,the detection rate was 86. 0%,320 cases were detected by MPR,the detection rate was 95. 5%,the differences were statistically significant (P<0. 05). The overall accuracy rate of Preoperative T staging by conventional axial and MPR images were 78. 3% and 89. 1% respectively, the difference was statistically significant. The overall accuracy rate of preoperative N staging by two methods were 73. 6% and 82. 3%,respectively,the difference was not significant. Conclusion Application of dual source spiral CT multiplanar reconstruction can significantly improve the detection rate of gastric cancer,and the average accuracy rate of preopera-tive TNM staging for gastric cancer is 84. 6%.
8.Expression of DNA methyltransferases in hilar cholangiocarcinoma and its clinical significance
Xiaoyu YIN ; Jianxin PENG ; Kunsong ZHANG ; Xun HOU ; Dong CHEN ; Lijian LIANG
Chinese Journal of Digestive Surgery 2012;11(5):467-470
Objective To investigate the expression of DNA methyltransferases ( DNMTs) in hilar cholangiocarcinoma and its clinical significance.Methods A total of 150 samples of cholangetic tissues were collected from 111 patients with hilar cholangiocarcinoma ( cholangiocarcinoma group) and 39 patients with choledochocele ( control group) at the First Affiliated Hospital of Sun Yat-Sen University from April 1997 to March 2007.A tissue chip containing the samples of hilar cholangiocarcinoma and choledochocele was prepared.Expressions of DNMT1,DNMT3a and DNMT3b were detected by the immunohistochemical staining. Differences in the protein expressions of DNMTs in the cholangiocarcinoma group and the control group were compared,and the correlation between DNMTs protein expressions and clinicopathological features was analyzed.All data were analyzed by using the chi-square test or Fisher exact probability.The survival curve was drawn by using the Kaplan-Meier method and the survival rate was compared by using the Log-rank test.Results The rates of high protein expressions of DNMT1 and DNMT3b were 54.1% (60/111) and 47.7% (53/111) in the cholangiocarcinoma group, which were significantly higher than 28.2% ( 11/39) and 23.1% ( 9/39) in the control group ( x2 =7.740,7.240,P <0.05). The high protein expression of DNMT1 was correlated with-the Bismuth-Corlette classification and T staging of the tumor ( x2 =12.200, 17.800,P <0.05) ; there was no significant difference in the high protein expressions of DNMT3a in the cholangiocarcinoma group and the control group ( x2 =3.370.P >0.05 ) ; while the high protein expressions of DNMT3b was correlated with the Bismuth-Corlette classification (x2 =8.300,P < 0.05 ),but not with the T staging. Sixty-six patients received hilar cholangiocarcinoma resection,and 42 of them were followed up.The median postoperative survival time of patients with low protein expression of DNMT1 was 23.9 months,which was significantly longer than 11.8 months of patients with high protein expression of DNMT1 (x2 =3.980,P < 0.05).Conclusions DNMT1 and DNMT3b with high protein expression might play important roles in the carcinogenesis and development of hilar cholangiocarcinoma.There is an obvious relationship between the expression of DNMT1 and postoperative survival time of patients with hilar cholangiocarcinoma,and DNMT1 might be a valuable prognostic factor for hilar cholangiocarcinoma.
9.Inducing effect of IL-1βabout the release of HMGB1 in human bronchial epithelial cells
Xiaoyu TAN ; Changchun HOU ; Junjian CHEN ; Yu LI ; Tangjuan LIU ; Hongjiang ZHOU
Chongqing Medicine 2015;(2):151-154
Objective To investigate the effect of IL‐1βon the expression and release of high mobility group box 1 (HMGB1) from normal human bronchiolar epithelial cell (HBE) .Methods HBE135‐E6E7 was developed and methyltetrazolium (MTT) as‐say was used to assess the viability of HBE cell line under different concentration of IL‐1β.The mRNA expression of HMGB1 in HBE after stimulating with IL‐1βwere determined by Real‐time PCR;the level and location of HMGB1 in the cytoplasm ,nucleus and culture medium of HBE after stimulating with IL‐1β were detected by Western blot and ELISA assay .The expression and translocation of HMGB1 of HBE after stimulating with IL‐1β were detected by Immunofluorescence .Results 0 .1 ,1 .0 ,10 .0 ng/mL IL‐1βdid not influence the cell viability of HBE ;IL‐1β increased the mRNA expression of HMGB1 in HBE in does‐and time‐dependent manner and increased the protein expression of HMGB1 in HBE .In comparison with control group ,the levels of HMGB1 in the culture medium significantly increased after stimulation with IL‐1βat 1 .0 ,10 .0 ng/mL for 24 h(P<0 .05)in the dose dependent experiments;and in time dependent experiments ,10 .0 ng/mL IL‐1βsignificantly increased HMGB1 level in the cul‐ture medium after stimulation for both 12 h and 24 h (P< 0 .05) .After stimulation with IL‐1β (10 .0 ng/mL ) for 24 h ,the HMGB1 expression in cytoplasm significantly increased in plasmosin and decreased in nucleus .HMGB1 translocated from nuclei to cytoplasm after stimulation with IL‐1β(10 .0 ng/mL ) for 24 h .Conclusion IL‐1βcould induce HMGB1 expression and release in human bronchial epithelial cells ,which indicates that HMGB1 may involve in IL‐1β‐mediated chronic airway inflammation .
10.Evaluation of genetic diversity of Dendrobium officinale wild populations based on RAMP markers.
Jie SHEN ; Huijun XU ; Yinghui YUAN ; Li HAN ; Beiwei HOU ; Xiaoyu DING
Acta Pharmaceutica Sinica 2011;46(9):1156-60
Random amplified microsatellite polymorphism (RAMP) markers were used to access the genetic diversity among 112 samples of nine populations of Dendrobium officinale Kimura et Migo. Using 16 informative primers, 123 bands were amplified and 86 (69.92%) were polymorphic. The polymorphic bands from three to eight could be detected for each RAMP primer, with a mean of 5, indicating abundant genetic diversity among populations. Genetic similarity coefficients ranged from 0.250 to 0.813. UPGMA dendrogram illustrated 9 populations clustered into 3 groups, and the cluster pattern showed correlation with the locations of the D. officinale populations. These results were supported by the previous conclusions that were achieved by other molecular markers, and RAMP is proved to be effective for evaluating the genetic diversity of wild populations of Dendrobium officinale.