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16"different" adhesion, invasion, growth, is the key difference between eutopic endometrium, eutopic endometrium of endometriosis is a decisive factor of occurrence disease and endometriosis recurrence.Chinese medicine treatment of endometriosis in clinical advantages unique increasingly medical attention. In the experiment of tanshinone IIA in endometriosis patients, to study its effect on endometrium adhesion. Materials and methods
1. Patients with endometriosis eutopic endometrial cells and cells of normal endometrium culture
2. Effects of Tanshinone IIA in eutopic endometrium adhesion index of endometriosis
3. Refer to method to culture cells of Ryan, we successfully established Eutopic endometrium cell of endometriosis in vitro model.
4. We put different concentrations of Tanshinone IIA on the Eutopic endometrium cell of endometriosis, Using the western blot method to test protein content of expression of E-cadherin, CD44V6, ICAM-1.
Results and discussion
1. This study confirmed that training of the two groups of different period of Eutopic endometrium and training of the two groups of different seasons both had statistical significance. P<0.05.But training of the two groups of different ways of getting material of Eutopic endometrium had no statistical significance, P>0.050
2. Protein of E-cadherin, CD44V6, ICAM-1 in Control group are high expression, comparing with this group, High dose group of Tanshinone IIA had lower expression, p<0.01 ; Middle dose group and low dose group of Tanshinone IIA had lowest expression, p<0.05.
3. This study successfully established the cell culture model in vitro of patients with endometriosis eutopic endometrial cells of the uterus.
4. The experimental results showed that IIA could increase the content of tanshinone endometriosis eutopic endometrial cell adhesion protein E-cadherin, CD44v6 and ICAM-1 in the occurrence and development of uterine, may prevent the endometriosis.
References:
[1]. Le Jie. Obstetrics and Gynecology [M]. 7 edition. Beijing: People's Medical Publishing house,2008:325-331.
[2].Cramer DW, Missmer SA. The ePidemiology of endometriosis [J].Ann N Y Acad Sei.2002;955:11-22.
[3]. Lang Jinghe, and clinical study of endometriosis foundation problems [J]. Chinese Journal of Practical Gynecology and obstetrics.2002,18 (3): 129-130.
The role of CAPN5 in the pathogenesis of endometriosis
Huifang Cong, Tianchan Zhang The Second Affiliated Hospital of Heilongjiang University Of Chinese Medicine,haerbin,china
Abstracts: To establish endometriosis endometrial glandular epithelial cells and stromal cells in vitro co-culture model. To determine and compare expression of Calpain5 in each group with Western blot. Identification of endometrial cell by immunocytochemistry. The results were confirmed using western blot, a decrease in Calpain5 protein, respectively, in endometriosis compared with endometrium obtained from controls (P<0.05). Ecreased apoptosis in the eutopic endometrium of women with endometriosis compared with endometrium obtained from controls implicate that, aberrant expression of CAPN5 is associated with endometriosis
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Keywords: endometriosis, Western blot, CAPN5
Introduction
Endometriosis is a common disease of fertile women, defined by the presence of ectopic endometrial glands and stroma. The incidence of endometriosis is rising recently, the disease performance diversity, so far the pathogenesis of it is unclear. Calpain5 is a member of the cytoplasmic cysteine protease family, a group of Ca++ regulated enzymes. Calpains in general, and in particular Calpain5, have been implicated in multiple cellular processes including cellular differentiation and apoptosis.
Materials and Methods
Tissue collection
10 endometrial biopsies were obtained from women with hysteromyoma as controls. Histologically confirmed biopsies of endometriosis from these wih endometriosis were obtained from 16 women.All women were in the reproductive age were not using hormonal medication within 3 months. Approvals were obtained from four hospitals of Medicine Human Investigations Committee.
Materials
PMSF(100mm), TEMEMD, DMSO, DMEM/F12, enzyme, ELIASA, centrifugal, Su perClean bench, Gel Imaging et al.
Cell culture
Eutopic endometrium were obtained from 16women with endometriosis as a group, 10 eutopic endometrium were obtained from women without endometriosis as controls. Isolated and culture primary glandular epithelial cells and stromal cells.Each was supplemented with 15% charcoal-stripped calf serum,1.5% penicillin/streptomycin .Placed in 37 ° C , 5 % CO2 cell incubator cultured overnight.
Immunohistrchemistry
Identification of endometrial cell by immunocytochemistry under the instructions.
Western blot
Endometriosis group to cultured cells , the normal control group cells extraction of total cellular protein in the culture medium adherence and drug intervention , measurement of protein concentration . Cleaning irrigation plastic glass , were added with a good 10% separating gel , 4% stacking gel plug the hole on the comb into the electrophoresis tank , add enough liquid electrophoresis .The sample run on a gel , transferred to a membrane , PVDF membrane was blocked , an antibody incubation , the secondary antibody incubation , exposure to the chemiluminescence of the A and B reagent , tableting , a developing fixing.Gel image processing system analysis target strip barren density value of the gray scale value of the target band and the internal reference strip than the relative expression levels of the target protein .
Statistics
PSS17.0 software , and all data observation data results are expressed as mean± standard deviation (x ±s), one-way ANOVA and Independent samples t test for comparison between the multi- group and two groups. The difference was statistically significant when P <0.05 .
Results
Immunochemical identification
CK19 is anti-body cytokeratin for Glandular epithelial cells and vimentin for the stromal cells.After staining cytoplasm , rendering the brownish-yellow , blue-purple nucleus showed positive results .
Figure 1 Figure 2 Figure 1 , glands cytokeratin CK19 staining positive Figure 2 , stromal cells, vimentin Vim stainingpositive
Calpain5 epression was decreased in endometriosis
The endometriosis eutopic endometrial cells CAPN5 protein relative expression levels lower than the normal control group reign endometrial cells CAPN5 protein relative expression levels, Statistically meaningful (P<0.05).
Figure 3
Figure 3 Western the the Blot endometriosis and endometrial cells in the normal control group CAPN5 protein relative expression levels.1,3,5,7 for the normal control group .2,4,6,8 for endometriosis group .
Relative expression level of the normal control group the cell CAPN5 protein the mean value is set to 1and protein relative expression of endometrios in cells CAPN5 is 0.432 ± 0.096,two groups statistically significant (P<0.05) .
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Figure 4
Figure 4 , columnar expression of the normal control group and endometriosis cells endometrial cells CAPN5 protein results. Discussion
Endometriosis in women of childbearing age in the incidence rate of 10-15% , 30-50 % accompanied dysmenorrhea , 80% of patients with pelvic chronic pain , 50% with infertility[1] . With the increased incidence of endometriosis has become a veritable " modern disease " . Endometriosis and in-depth , but its incidence caused by lesions morphological diversity , internal specific pathogenesis of endometriosis is still not clear enough .With the the eutopic endometrium
decision proposed , for the study of the pathogenesis of endometriosis bring new ideas and direction of endometriosis occurs due to abnormal eutopic endometrium biological behavior expressed.
CAPN5 a neutral cysteinyl amino acid protease family , depending on the catalytic subunit , were classified as non -classical , is a calcium -dependent protease. Shows CAPN5 in man and rodents significant homology , there is 87% DNA and 95% of the same protein sequence , while , CAPN5 Caenorhabditis TRA-3 homology shows that this protein gene evolution reproductive function , in the current report , not yet see CAPN5 disease-related [3]D CAPN5 expression in multiple tissues of the human body , including the nervous system , uterus , placenta , etc. [4-5]Recently, academic studies show that CAPN5 expression go in both glandular epithelial cells and stromal cells , and its expression can be sustained throughout the menstrual cycle , and showed increased expression levels in the secretory phase.In1933 CAPNs was first reported to induce apoptosis , the researchers pointed out the , CAPNs participate in certain stimulation caused by the process of apoptosis , but the specific apoptotic mechanism mediated pathway remains unclear.The CAPN5 as a CAPNs family can participate in the activities of a variety of cells , including cell differentiation and apoptosis[6]. CAPN5 mediated pathway of apoptosis perform protein caspase3, and can be activated to induce cell apoptosis can also be induced pathway mediated by mitochondria ,cells-resulting in apoptosis of isletcells [7-10].
Research and investigate the expression levels of CAPN5 eutopic endometrium of endometriosis patients , is of positive significance for further discussion to address the endometrium of endometriosis pathogenesis and treatment strategies. The experimental research study results showed that the Endometriosis women reign the endometrial cells CAPN5 protein , both at the level of gene regulation in protein expression levels are significantly lower than the normal control group of women, CAPN5 low level of expression , including may be of great significance to the development of endometriosis , endometriosis eutopic endometrial cell apoptosis in abnormal regulation , resulting in reduced apoptosis , in ectopic pelvic activity , survival The ability proliferative capacity strengthening , resulting Benbingfasheng .
In summary , apoptosis the protein CAPN5 because the abnormal regulation of the gene , resulting in expression levels were significantly lower in endometriosis eutopic endometrial cells , causing inhibition of apoptosis of endometrial cells , the cells in the long-term survival perhaps plays an important role in the etiology and pathogenesis of endometriosis . References
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[2] Jones RK,Searle RF,Bulmer J.Apoptosis and bcl-2 expression in normal human endometrium,endometriosis and adenomyosis.Hum Reprod[J].1998,13(12): 3496-3502.
[3] Syntichaki P,Xu K,Driscoll M,Tavernarakis N.Specific aspartyl and calpainproteases are required for neurodegeneration in C.elegans[J].Nature,2002,419: 939-944.
[4] Gonzalez A,Saez ME,Aragon MJ,Galan JJ,Vettori P,Molina L,Rubio C,Real LM,Ruiz A,Ramirez-Lorca R.Specific haplotypes of the CALPAIN-5 geneare associated with polycystic ovary syndrome[J] .HumReprod2006,21:943-951.
[5] Artus C,Maquarre E,Moubarak RS,Delettre C,Jasmin C,Susin SA,Robert-Lezenes J.CD44 ligation induces caspase-independent cell deathvia a novel calpain/AIF pathway in human erythroleukemia cells[J].Oncogene 2006,25:5741-5751.
[6] Koichi S,Kawabata Y,Sorimachi .Structure,activation and biology of Caplain[J]. Diabetes Care 2004,53:s12-s18.
[7] Kayisli UA,Berkkanoglu M,Zhang L,Kizilay G,Arici A.The broadspectrumchemokine inhibitor NR58-3.14.3suppresses theimplantation and survivalof human endometrial implants in the nude mice endometriosis model.Reprod Sci[J].2007,14:825-835.
[8] Lee WK,Abouhamed M,Thevenod F.Caspase-dependent and-independentpathways for cadmium-induced apoptosis in cultured kidney proximaltubule cells.Am J Physiol Renal Physiol[J] .2006,291 :F823-F832.
[9]Johnson JD,Han Z,Otani K,Ye H,Zhang Y,Wu H,Horikawa Y,Misler S,Bell GI,Polonsky KS:RyR2 and calpain10 delineate a novel apoptosis pathway in pancrestic islets.The Journal of biological chemistry[J] .2004,279(23):24794-24802.
[10] Arrington DD,Van Vleet TR,Schnellmann RG:Calpain10:a mitochondrial calpain and its role in calcium-induced mitochondrail dysfunction. Am J Physiol Cell Physiol [J]. 2006, 291 (6):C11 59-71.
Electroacupuncture for Treating ProstaticHyperplasia:A Randomized
Controlled Trial
Wang Chunying,Jin Ze,Jiang Shanshan
1 No.2 Hospital Affiliated to HeilongjiangUniversity of Traditional Chinese Medicine, Harbin
150001, P. R. China
Abstract:Objective: To assess the clinical effect of electroacupuncture for treating prostatic hyperplasia.Methods: Sixty patients with prostatic hyperplasia were randomized into a treatment group and a control group,30 in each. The treatment group was treated byelectroacupuncture, and the control was prescribed with orallytaken Jing Zhu Qian Lie Long Bi Tong Capsule. After treatment, the two groups were compared with each otherinterms of treatment result, international prostate symptom score (I-PSS), quality of life index (QLI), maximumflow rate (Qmax), and residual urine volume (RUV). Results: The total effective rate was 96.7% in the treatmentgroup versus 70.0% in the control, and thedifference was significant (P<0.05). Both groups had markedimprovement in I-PSS, QLI score, Qmax and RUV (P<0.05). Conclusion: By improving I-PSS, QLI, Qmaxand RUV,electroacupuncture is an effective way for treating prostatic hyperplasia.
Key Words:Electroacupuncture; Acupuncture Therapy; Prostatic Hyperplasia; Difficulty in Urination;Randomize Controlled Tiral
Benign prostatic hyperplasia is a common diseasein elderly males. According to the survey conductedby Gu[1] in 20 hospitals, 13.6% of the inpatients inurinary department were prostatic hyperplasia. Wetreated this disease by electroacupuncture andcompared the effect with medication. The report isnow given as follows
Materials and methods.
1 Clinical Data
1.1 Diagnosis criteria[2]
® Urinary symptoms such as urinary hesitancy,obstructed urine, urination frequency worse at night,urine leakage afterwards, distending pain in the lowerbelly, but absence of irritating pain in the urinary tract;© digital rectal examination finds prostateenlargement, and dullness topercussion in lower belly;©diagnosis confirmed by ultrasonic scanning; ©urinary infection,urinary tract stone and prostatecancer should be excluded.
1.2 Inclusion criteria
® Conformed to the diagnosis criteria; ® willingto participate in this randomized controlled clinicalstudy and sign the informed consent form.
1.3 Exclusion criteria
® Allergic to the medication used in the presentstudy; ® accompanied by mental health problem,liver or kidney dysfunction, hepatopathy, or infectiousdiseases; © patients who adopted other treatmentmethods in recent 2 weeks.
1.4 Termination criteria
