3.Irritation of a new specimen preservation solution on the skin,eye of rabbits and its effect for specimens preser-vation
Xindong LIANG ; Wenxia MA ; You ZHANG ; Zheng NIE
Journal of Regional Anatomy and Operative Surgery 2015;(4):376-378,379
Objective To conduct the preliminary toxicology tests and evaluate the preservation effect on a new specimen preservation solution in order to provide scientific basis for its application security. Methods Twenty-four New Zealand white rabbits were randomly di-vided into the normal saline group, the formalin preservation group,the mixture group,and the new preservation solution group. Recorded the irritation of different solution on the skin,eye of rabbits,the effect of liver sample preservation in different solution,and the evaporation rate of these solution. And to find a set of preservative solution which is more suitable for preservation of specimens and can reduce the cost. Results The new preservation solution is stimulus on rabbit skin and eye,and it is stronger than the formaldehyde group. Compared with the other three groups,the mixture group has the best preservation effect, and the new preservation solution group has a better preservation effect than the formalin preservation solution group. Conclusion The 1∶1 mixture solution of the new preservation solution and formaldehyde is more suitable preservation solution and it can reduce the costs.
4.COMMD7 inhibits migration and invasion in liver cancer stem cells via regulating mesenchymal-epithelial transition
Ye TAN ; Nan YOU ; Lu ZHENG ; Xiaobing HUANG ; Liang WANG ; Ke WU ; Changlin DENG ; Jing LI
Journal of Third Military Medical University 2017;39(17):1691-1695
Objective To determine the effect of COMMD7 inhibition on invasion and migration in liver cancer stem cells (LCSCs),and investigate the possible mechanism.Methods After LCSCs were infected by shRNA lentiviral vectors of COMMD7,adhesion assay and Transwell assay were used to detect the invasion and migration,and phalloidin staining was employed to observe the morphological changes.Western blotting was adopted to measure the expression of E-cadherin,N-cadherin and Vimentin.Results COMMD7 knockdown significantly inhibited the invasion and migration of LCSCs.The relative cell quantity of adhesion was 1.00 ± 0.12 and 2.35 ± 0.20 respectively in control cells and infected cells,suggesting there were significantly more adhesive cells in the infected group (P < 0.05).The relative cell quantity per visual field of migration was 1.00 ±0.04 and 0.24±0.03,and that of invasion was 1.00 ±0.05 and 0.24 ±0.04 respectively in the control cells and infected cells,and there were significantly less invasive and migrated cells in the infected group (P <0.05).What's more,COMMD7 knockdown also induced some morphological changes of cells corresponding to the weakened abilities of migration and invasion.All the changes above were associated with up-regulation of E-cadherin (P < 0.05) and down-regulation of N-cadherin and Vimentin (P <0.05),the molecules related to mesenchymal-epithelial transition (MET).Conclusion COMMD7 knockdown inhibits the invasion and migration in LCSCs,which may be through its regulation on the MET course.
5.Establishment and application of a multiplex PCR method in Staphylococcus aureus、Pseudomonas aeruginosa and Klebsiella pneumoniae
Yanbo ZHU ; Long ZHENG ; Hongyu YOU ; Xuan WANG ; Xiaoliang LIANG ; Jianmin LIU ; Dongming ZHANG ; Weiguang LIAN ; Yanning LI ; Junxia WANG
Chinese Journal of Comparative Medicine 2017;27(5):94-98
Objective Aiming at detecting Staphylococcus aureus、Pseudomonas aeruginosa and Klebsiella pneumoniae in laboratory animals,the paper provides a rapid,sensitive and simple test method.Methods According to Staphylococcus aureus nuc gene,Pseudomonas aeruginosa LasI gene,Klebsiella pneumonia PhoE gene and general 16S rRNA gene, designed specific primers;Through the optimization of multiplex PCR primer concentrations and annealing temperature, the specificity and sensitivity of detection, establishing multiplex PCR system.Application of the PCR system test specimens of artificial infections and experiment animal feces is compared with traditional test method.Results Multiplex PCR amplification of Staphylococcus aureus (153 bp), Pseudomonas aeruginosa (600 bp) with Klebsiella pneumoniae (368 bp) and general (520 bp).The multiplex sensitivity for the purpose of 10pg, specificity of detection was not detected from other pathogens.Application of establishing multiplex PCR system to detect the artificial positive samples, and detect 1 Pseudomonas aeruginosa positive case in 76 fecals.Conclusions This paper established the multiplex PCR method which has the advantages of specific,sensitive,simple and rapid, and provides a reliable way for rapid test in laboratory animals microbiology.
6.Establishment and application of a multiplex PCR assay for four pathogens in laboratory animals
Yanbo ZHU ; Zengnian XU ; Shijin WEI ; Long ZHENG ; Hongyu YOU ; Yurong MENG ; Fuying LIU ; Xiaoliang LIANG ; Junxia WANG
Chinese Journal of Comparative Medicine 2017;27(8):80-84
The aim of this study is to establish a multiplex polymerase chain raction (PCR) to identify of four kinds of laboratory animal pathogens: Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae and Klebsiella pneumoniae.Methods Specific primers were designed based on GenBank data.The multiplex PCR system was established through optimization of multiple PCR and detection of its specificity and sensitivity.This technique was used to test artificially infected samples and tracheal secretions of experimental animals (rat, mouse, guinea pig, rabbit, hamster), and comparing the detection results by this method and traditional detection test.Results Target bands of Pasteurella multocida (356 bp), Bordetella bronchiseptica (237 bp), Mycoplasma pneumoniae (266 bp), and Klebsiella pneumoniae (142 bp) were obtained, with a detection sensitivity of Klebsiella pneumoniae of 10 pg, and that of Pasteurella multocida, Bordetella bronchiseptica and Mycoplasma pneumoniae of 1 pg by this newly developed multiplex PCR assay.No target bands were observed from the non-specific pathogens of artificially infected samples.The tracheal secretions taken from 45 experimental animals (mice and rabbits) were tested with this new PCR assay, among which 15 cases of Klebsiella pneumonia and 9 cases of Pasteurella multocida were detected as positive, while all the results of traditional method and serological test were negative.Conclusions A simple, rapid, specific and highly sensitive multiplex PCR system has been successfully established.It is valuable for detection of Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae, and Klebsiella pneumoniae in laboratory animals.
7.Design and Realization of Independent Calling Device for Patients with Amyotrophic Lateral Sclerosis
Xiao-qi ZHU ; You-jun LI ; Wei-jie TAO ; Yu-hui WEI ; Liang ZHENG ; Guo-dong FENG ; Jue WANG
Chinese Journal of Rehabilitation Theory and Practice 2015;21(7):745-747
Objective To design a device for the patients with amyotrophic lateral sclerosis (ALS), who are disable to speak and move, to call for the nurses independently when necessary. Methods The electroencephalographic signals were collected and processed with TGAM, and the extracted attention characteristic values were transmitted to a computer with Bluetooth. The loudspeaker would call the nurses when the attention characteristic value exceeds the normal range. Results In the testing process of 20 participants in 10 tests, the success rate was 81.5%, and the average misjudged frequence was 0.2 within 20 min. Conclusion The device can be used in the nursing of the patients with ALS to meet the needs of daily nursing work.
8.Diagnostic value of urodynamic parameters multianalysis in the evaluation of bladder outlet obstruction resulting from benign prostatic hyperplasia.
Yue-you LIANG ; Ming-xin CAO ; Yu-ping DAI ; Wei-jie LIANG ; Ke-li ZHENG
National Journal of Andrology 2006;12(9):818-821
OBJECTIVETo construct a function model that can be used in the diagnosis bladder outlet obstruction (BOO) resulting from benign prostatic hyperplasia, and to develop a diagram allowing the judgement of bladder outlet for patients with different detrusor contractility, especially with impaired one.
METHODSUrodynamic and clinical data of 131 men were analyzed retrospectively. By Logistic analysis, a function model was constructed. Based on the model, a diagram allowing the evaluation of bladder outlet was drawn. The cutoff point for diagnosing BOO with the function model and the curve was confirmed by ROC curve analysis.
RESULTSThe function model (BOOI) was obtained by the formula 5.03 x residual fraction + 0.04 x PdetatQmax - 0.20 x Qmax - 0.91 + alpha (alpha = 0 for those with low pressure-low flow on P-FS, alpha = 1.42 for high pressure-low flow, alpha = -7.30 for high pressure-high flow). The cutoff point for BOOI diagnosing BOO was 0.36. When validated, the sensitivity, specificity, positive predictive value, and negative predictive value were 85.7%, 91.7%, 96.0% and 73.3% respectively.
CONCLUSIONThe BOOI, with an easy calculation mode, could predict the probability of BOO. The sensitivity and specificity of the criterion for the diagnosis of BOO were satisfactory. The curve we drew could help to differentiate the obstructed men with low pressure-low flow and thus benefit them by surgical relief of their obstruction.
Aged ; Aged, 80 and over ; Humans ; Logistic Models ; Male ; Middle Aged ; Prostatic Hyperplasia ; complications ; ROC Curve ; Retrospective Studies ; Urinary Bladder Neck Obstruction ; diagnosis ; etiology ; Urodynamics
9.Persistent Muellerian duct syndrome with transverse testicular ectopia.
Yue-You LIANG ; Fu-Fu ZHENG ; Yu-Ping DAI ; Ke-Li ZHENG ; Jie-Xue ZHOU
Asian Journal of Andrology 2006;8(6):745-747
Persistent Muellerian duct syndrome (PMDS) is a rare form of male pseudohermaphrodism without the feature of ambiguous genitalia. We present a case of PMDS with transverse testicular ectopia (TTE).
Abnormalities, Multiple
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Adult
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Disorders of Sex Development
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pathology
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surgery
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Hernia, Inguinal
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surgery
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Humans
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Male
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Mullerian Ducts
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abnormalities
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surgery
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Testicular Hydrocele
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surgery
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Testis
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abnormalities
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surgery
10.Correlation of expression of RhoC with invasiveness of breast cancer cells in vitro.
Zheng YUAN ; Jling SU ; Jiang-Feng YOU ; Jie-Liang WANG ; Xiang-Lin CUI ; Jie ZHENG
Chinese Journal of Oncology 2007;29(12):898-903
OBJECTIVETo investigate the expression of RhoC in breast cancer cells with different metastatic potential and its correlation with invasiveness.
METHODSExpression of RhoC mRNA and protein in human breast cancer cells MCF-7 with low metastatic potential and MDA-MB-231 with high metastatic potential was detected by RT-PCR, Western blot, immunohistochemistry and immunofluorescence staining. Eukaryotic expression plasmids of RhoC were constructed and transfected into MCF-7 cells. The biological effects were observed, including in vitro invasion by Boyden charmber assay, motility by would healing assay, alteration of microfilament network by TRTIC-phalloidin staining and expression of p-Akt by Western blot assay.
RESULTSThe expression levels of RhoC mRNA and protein varied in the two different metastatic breast cancer cell lines. RhoC was significantly up-regulated in the highly metastatic cells in comparison to the weakly metastatic counterpart (P < 0.01). As shown by Boyden charmber assay, the invasive capacity of transfected cells overexpressing RhoC was significantly promoted as reflected by more penetrating cells (56.88 +/- 4.18) than that of the antisense transcripts (23.12 +/- 3.22), the negative (23.77 +/- 3.64) and blank controls (28.44 +/- 2.48). Further study by would healing assay indicated that cells overexpressing RhoC were more motile in actin-based active movement. The wound healing ratio after 24 h of the sense transcripts, antisense transcripts, negative controls and blank controls was 58.28% +/- 2.14%, 22.36% +/- 2.73%, 28.23% +/- 2.62%, 30.18% +/- 2.86%, respectively. The TRITC-phalloidin staining revealed less actin filament bundles and a reorganized cytoskeleton within the sense transcripts. In addition, p-Akt expression level was upregulated in the sense transcripts.
CONCLUSIONRhoC overexpression may promote the invasive capacity of human breast cancer cells in vitro and its expression level is positively correlated with the metastatic capacity of those cells. So RhoC may be a potential target in the development of a novel strategy for treating metastasis of breast cancer.
Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Neoplasm Invasiveness ; Phosphorylation ; Plasmids ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; metabolism ; Transfection ; Up-Regulation ; rho GTP-Binding Proteins ; genetics ; metabolism ; rhoC GTP-Binding Protein