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Objective To investigate the expression of NLRP3 inflammatory body in the process of liver fibrosis in a rat model of common bile duct ligation (BDL) and the association of NLRP3 inflammatory body with liver fibrosis. Methods A total of 65 Sprague-Dawley rats were randomly divided into sham-operation group with 15 rats and BDL model group with 50 rats. On days 3, 7, 14, 21, and 28, 10 rats in the model group and 3 rats in the sham-operation group were sacrificed. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), direct bilirubin (DBil), total bilirubin (TBil), total bile acid (TBA), and alkaline phosphatase (ALP) were measured, and HE staining, Masson staining, and sirius red-picric acid staining were performed for liver tissue to evaluate liver fibrosis degree. Immunohistochemistry was used to measure the expression levels of alpha-smooth muscle actin (α-SMA) and transforming growth factor-β1 (TGF-β1) in liver tissue, Western blot and qRT-PCR were used to measure the expression level of NLRP3 inflammatory body, and ELISA was used to measure the level of the inflammatory factor interleukin-1β (IL-1β) in liver tissue. An analysis of variance was used for comparison of continuous data between groups, and the least significant difference t -test was used for further comparison between two groups. Results Compared with the sham-operation group, the BDL model group had significant increases in the serum levels of ALT, AST, DBil, TBil, TBA, and ALP (all P < 0.05) and the level of IL-1β in liver tissue ( P < 0.05), which reached the highest level on day 3 and then decreased. Compared with the sham-operation group over time, the BDL group had a significant increase in liver fibrosis score ( P < 0.05); immunohistochemistry showed gradual increases in the expression of SMA-α and TGF-β1 ( P < 0.05), and Western blot and qRT-PCR showed a gradual increase in the protein expression of NLRP3 inflammatory body in liver tissue ( P < 0.05), which remained stable after day 14. Conclusion Liver injury exists persistently in a rat model of BDL, and liver histopathology shows the dynamic evolution of hepatitis, liver fibrosis, and liver cirrhosis. NLRP3 inflammatory body is in a state of continuous activation and may play an important role in the process of liver fibrosis.
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Objective To observe the effect of saikosaponin-d on transcriptional activation of estrogen receptor in rat hepatic stellate cells(HSC-T6), and to explore its pharmacological mechanism. Methods The rat HSC-T6 were cultured in vitro for the study. After transient transfection of HSC-T6 with the ER-specific reporter gene ERE-tk-Luc by liposome, the effect of saikosaponin-d and estradiol on luciferase activity was observed by Dual-Luciferase Reporter Assay System under the conditions of various drug concentrations, various treatment time and addition of estrogen receptor inhibitor ICI182. 780. Results When the concentrations of saikosaponin-d and estradiol were in the range of 0.01-5 μmol/L, 0.01-1 μmol/L respectively, luciferase activity was increased in a dose-dependent manner . Luciferase activity arrived the highest when saikosaponin-d concentration was 5 μmol/L and estradiol concentration was 1 μmol/L, but the effect was weakening when estradiol concentration reached 5μmol/L. In respect of the effect of treatment time, when HSC-T6 were separately treated with 5μmol/L of saikosaponin-d and 1 μmol/L of estradiol for 24 h, the luciferase activity was the highest. And ICI182.780 could significantly inhibit the induction of saikosaponin-d and estradiol for luciferase activity. Conclusion Saikosaponin-d has an effect on promoting the transcriptional activation of estrogen receptor in HSC-T6.
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Purpose To investigate the relationship between fetal lateral ventricle dilatation complicated with malformation and chromosome abnormalities,so as to provide reference for clinical consultation.Materials and Methods The ultrasound images,karyotype analysis and high resolution microarray comparative genomic hybridization (aCGH) results of 150 fetuses with lateral ventricular dilatation diagnosed by ultrasound were analyzed retrospectively.Results Among 150 lateral ventricular dilatation fetuses,81 cases were isolated lateral ventricle dilatation,30 cases were found complicated with fetal ultrasound soft index,22 cases with other CNS malformations and 17 cases with other malformations.Karyotype analysis of the above 4 groups showed 13 cases of abnormal karyotypes and 15 cases ofaCGH abnormalities.There was statistical significant difference (P＜0.05) in each group of abnormal chromosome and aCGH detection rate,in which the softer marker's group had a significantly higher rate than the isolated ventriculomegaly's group in abnormal chromosome and aCGH (P＜0.05).There was no statistical significant difference in any other groups (P＞0.05).Conclusion Fetal systems should be carefully examined when prenatal ultrasound reveals lateral ventricular dilatation.The probability of abnormal chromosome increases significantly when complicated with fetal ultrasound soft index or other structural abnormalities.Chromosomal abnormalities need to be excluded and regular ultrasound follow-up is necessary in fetuses with isolated lateral ventricles.
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s:Objective Discuss the application of CTA for renal vascular control in laparoscopic renal transplantation of relative renal donors. Methods 83 cases of relative renal transplantation donors which are completed in our department during March 2007 to Junuary 2012 were divided into 3 Groups. There were 35 cases in group A ( laparoscopic nephrectomy with preoperative CTA examination) , 15 cases in group B ( laparoscopic nephrectomy without preoperative CTA examination) , and 33 cases group C ( conventional nephrectomy) . In group A, the location and quantity of renal arteries and veins were observed with preoperative CTA to make plan for vascular control. And the oper-ation time, amount of bleeding, vascular injury cases, the renal warm ischemia time in the graft, mean time of postoperative hospitalization were compared with those of group B and group C. Results 5 cases of renal vascular anomalies were observed in Group A, including 2 cases of single accessory renal artery, 2 cases of early branch of renal artery and 1 case of double renal veins. And all of them were intraoperatively comfirmed. 3 cases of renal vascular anomalies were observed in group B, including 1 case of renal arteriovenous ectopic, 1 case of injury in the accessory renal artery with diameter of 0. 6 cm, and it were given artery elongation with inferior epigastric artery postoperatively, and 1 case of renal vein injure during blocking lumbar vein with Hemolock which were repaired postoperatively. 5 cases of renal vascular anomalies were observed in group C, including 1 case of single accessory renal artery, 3 cases of early branch of renal artery, and 1 case of accessory renal artery with the diameter of 0. 1cm in the right renal was injured during surgery. The operation time, amount of bleeding, cases with vascular injury , renal warm ischemia time of donor , and the mean time of postoperative hospitalization in group A were superior to those in group B and group C. Conclusion CTA is a simple noninvasive imaging method, which can provide the details of the renal vascular preop-eratively to ensure the safety of patients during surgery and improve survival quality of donor renal, and it is of great advance in laparoscopic renal transplantation of living-related renal donors.
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This article describes a novel Multifunctional and Transparent Urinary System Model (MTUSM), which can be applied to anatomy teaching, operational training of clinical skills as well as simulated experiments in vitro. This model covers kidneys, ureters, bladder, prostate, male and female urethra, bracket and pedestal, etc. Based on human anatomy structure and parameters, MTUSM consists of two transparent layers i. e. transparent organic glass external layer, which constraints the internal layer and maintains shape of the model, and transparent silica gel internal layer, which possesses perfect elasticity and deformability. It is obvious that this model is preferable in simulating the structure of human urinary system by applying hierarchical fabrication. Meanwhile, the transparent design, which makes the inner structure, internal operations and experiments visual, facilitates teaching instruction and understanding. With the advantages of simple making, high-findelity, unique structure and multiple functions, this model will have a broad application prospect and great practical value.
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Female , Humans , Male , Kidney , Models, Anatomic , Models, Biological , Prostate , Ureter , Urethra , Urinary Bladder , Urogenital SystemABSTRACT
PURPOSE: This study was conducted to investigate the small double-stranded RNA (dsRNA) mediated anti-tumor effects of p21(WAF1/ClP1) (p21) transcriptional activation in vitro in the human glioma SHG-44 cell line. MATERIALS AND METHODS: Human glioma SHG-44 cells were transfected with dsRNA using LipofectAMINE 2000 transfection reagent. Real-time PCR and Western blot analysis were conducted to detect p21 and survivin mRNA and protein levels, respectively. Cell proliferation was examined by MTT assay. Cell cycle distribution and apoptosis were detected by flow-cytometric analysis. RESULTS: We found that dsRNA targeting p21 promoter (dsP21) significantly induced the expression of p21 at transcription and protein levels, and reduced the expression of survivin. AS well, dsP21 transcription significantly inhibited human glioma SHG-44 cell proliferation. Analysis of cell cycle distribution revealed that dsP21 transfection increased accumulation of cells in the G0/G1 phase and reduced accumulation of cells in the S phase. Further analysis revealed that dsP21 transcription led to an increase in both early and late stages of apoptosis in human glioma SHG-44 cells. CONCLUSION: In the present study, P21 activation by RNA-induced gene activation (RNAa) induced anti-tumor activity in vitro in a human glioma SHG-44 cell line. The results suggested that RNAa could be used for human glioma treatment by targeted activation of tumor suppressor genes.
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Humans , Apoptosis , Blotting, Western , Cell Cycle , Cell Line , Cell Proliferation , Genes, Tumor Suppressor , Glioma , Methods , Real-Time Polymerase Chain Reaction , RNA, Double-Stranded , RNA, Messenger , S Phase , Transcriptional Activation , TransfectionABSTRACT
Objective The purification methods of the exosomes derived form T cells were established in order to get high quan-tity exosomes .Methods Exosomes from T cells culture supernatants were purified by ExoQuick Precipitation ,ultrafiltration and sucrose gradient centrifugation ,differential ultracentrifugation ,and confirmed via using transmission electron microscopy .The pro-tein expression of the exosomes were analyzed by SDS-PAGE electrophoresis .Western blotting was used to test the expression of IL-2 .Results The protein concentration of the exosomes purified through ExoQuick Precipitation ,ultrafiltration and sucrose gradi-ent centrifugation were higher than through differential ultracentrifugation (P<0 .05) .SDS-PAGE displayed the difference among the exosome purified by three methods .Three kinds of exosomes all expressed IL-2 .Conclusion ExoQuick Precipitation ,ultrafiltra-tion and sucrose gradient centrifugation technique can obtain high purity and complete exosome sample .
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Objective To investigate the predictive value of spiral CT in composition changes of pediatric urinary.Methods A total of 25 pediatric patients with urinary stones were investigated.Eighteen patients with renal stones were stratified into two groups:an alkalization therapy alone group ( n =9 ) and a comprehensive therapy group (n =9).Flame atomic absorption spectrum (AAS) was employed to measure calcium level of the pediatric urinary stones.Spiral CT was employed to measure the peak CT number in vitro of all the pediatric urinary stones and 61 adult urinary stones,which served as controls.Results All pediatric urinary stones contained calcium ( 0.11％ - 26.30％ ).A positive correlation was observed between the CT number of pediatric urinary stone and its stone calcium level ( r =0.855,P ＜ 0.01 ).Compared to the alkalization therapy alone group,the CT number and stone calcium level of pediatricrenal stones in the comprehensive therapy group were significant higher (stone CT number:162 ± 60 HU VS.783 ±476 HU,P ＜ 0.01 ; stone calcium level:1.30 ± 1.52％ VS 19.83 ± 7.48％,P ＜ 0.01 ).Compared to ≤400 HU pediatric renal stones,＞ 400 HU renal stones contained more calcium (21.71 ± 5.27％,1.65 ±1.82％,P ＜ 0.01 ) and failed to dissolve by alkalization therapy alone ( x2 =11.455,P ＜ 0.01 ).Conclusions CT could be a predictive tool for composition changes of pediatric urinary stones.In clinical CT scanning setting,＞400 HU pediatric urinary stones probably will contain more calcium and not be suitable for alkalization therapy alone.
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BACKGROUND:Previous research has demonstrated that preoperative hormone therapy relieved clinicaI and pathological stages of prostatic carcinoma patients,reduced positive ratio of incisaI margin:however,it did not increase disease-free surviva J rate.Worth of preoperative hormone therapy for prostatic carcinoma remains unclear.OBJECTIVE:To assess the role of preoperative hormone therapy in localised prostate cancer.METHODS:Articles were searched from PubMed,Embase.Cochrane Library(No.4,2009),Chinese biomedicaf literature database,Chinese Scientific Journals full_text database,and Chinese Journal full-text database(published before October,2009).Randomized controlled trials which were diagnosed as Iocalised prostate cancer using pathology and cytology were included The sex and nationality were not limited.and patients did not have severe heart and lung diseases Prostatic carcinoma which occurred Later or repeatedly was excluded.Otherwise.randomized controlled trails which were coincidence with the inclusion criteria were also included.Review Manager 5.0 published by Cochrane was used for stafistical analysis.Overall survival rate.disease-free survivaI rate,positive surgical margin rate,positive lymph node rate.and seminal vesicle invasion rate were evaluated.RESULTS AND CONCLUSlON:Six randomized controlled trials totaling 1 027 padicipants were included.The result of meta analysis showed that there was no significant difference in overall surviva J rate[RR=0.94,95%C,(0.86,1.02)],disease-free survival rate[RR=1.02,95%CI(0.89,1.17)],positive lymph node rate IRR=0.86,95%CI(0.47,1.57)],and seminal vesicle invasion rate[RR=1.09,95%CI(0.74,1.59)]between single prostatectomy and preoperative hormone therapy plus prostatectomy,while there was significant difference in positive surgical margin rate[RR=0.46,95%CI(0.32,0.66)]This suggested that preoperative hormonal therapy prior to prostatectomy did not improve survival rate,positive lymph nodes rate and seminaI vesicle invasion rate:however.there was a significant reduction in the positive surgicaI margin rate.
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BACKGROUND: There are many in vitro selection method of sperm, and swim-up and density gradient centrifugation are commonly used. It remains unclear which method minimizes bad stimulation to the sperm and select sperm with high fertilization potential. OBJECTIVE: To evaluate the effectiveness of swim-up and gradient centrifugation preparation techniques on intrauterine insemination (IUI).METHODS: A computer-based online search of Cochrane Library, PubMed, EMBASE databases was performed, and some related journals were manually searched for related articles published between January 1966 and February 2009. The quality of included randomized controlled trials (RCT) and q-randomized trials (Q-RCT) was evaluated and Meta-analysis was conducted by the Cochrane Collaboration's software RevMan5.0. Experts.RESULTS AND CONCLUSION: A total of 6 studies were included, involving 4 RCTs and 2 Q-RCTs. A total of 486 patients (1 099 IUI cycles) were enrolled. The Meta-analysis indicated that there was no difference between swim-up and gradient centrifugation preparation techniques for the IUI in terms of cycle pregnancy rates [OR = 1.11, 95%CI(0.8,1.55)], miscarriage rates [OR = 0.31, 95%CI(0.09,1.04)], sperm count [the weight mean difference (WMD) =-0.89, 95%CI(-14.17,12.38)], sperm motility [WMD = -2.31, 95%CI(-7.27,2.65)]. There is insufficient evidence to confirm which is the best method in the two specific preparation techniques. The quality of study methods should be improved. And more measure parameters should be included when comparing it before or after treatment, such as sperm motility, sperm count, sperm function.
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RNA interference(RNAi), a highly conserved process of post-transcriptional gene silencing, can be induced by small interfering double-stranded RNA that mediate sequence-specific mRNA degradation. In the past several years, RNAi has been widely used as both an experimental tool to study mammalian gene function and a potential therapeutic approach to treat human diseases. In addition, some new proteins which involve in RNAi pathway have been characterized in mammalian cells. Here, we summarize the various molecules in RNAi, mechanism of action, and the current therapeutic applications in cancers.
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<p><b>OBJECTIVE</b>To compare the apoptosis and hemodynamic changes of the penile tissue in diabetic and healthy rats.</p><p><b>METHODS</b>Sixteen chronic diabetic and 10 matched normal control rats were injected with a single dose of streptozotocin (STZ) 65 mg/kg and the diabetic model established. Eight weeks later the rats were killed, their haemoglobin Alc( HbAlc) measured, and their penises harvested. TUNEL assay (in situ nick end-labeling) and flow cytometry were used to analyse the penile cells for apoptosis. The thickness and diameter of the arteries were measured.</p><p><b>RESULTS</b>TUNEL showed apoptosis in the chronic diabetic rats. Flow cytometric analysis showed the apoptotic peak in the penile cells of the chronic diabetic rats when compared with the controls (P < 0.05). There was significant difference between the diabetic and healthy rats. Morphological analysis revealed that the thickness of the artery wall increased while the artery diameter decreased in the diabetes group compared with the controls.</p><p><b>CONCLUSION</b>Diabetes can lead to increasing apoptosis of the penile tissue, cause a decrease in the penile artery diameter and the thickening of the penile artery wall.</p>
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Animals , Male , Rats , Apoptosis , Arteries , Pathology , Diabetes Mellitus, Experimental , Pathology , Erectile Dysfunction , Flow Cytometry , In Situ Nick-End Labeling , Penis , Rats, Wistar , StreptozocinABSTRACT
<p><b>OBJECTIVE</b>To observe the synergistic effects of paclitaxel and gemcitabine on prostate cancer cell line PC-3 in vitro.</p><p><b>METHODS</b>Cell morphology, MTU, flow cytometer and immunocytochemical method were used to observe the effects of 10(-6), 10(-7), 10(-8) mol/L paclitaxel and 10(-7), 10(-8), 10(-9) mol/L gemcitabine on prostate cancer cell line PC-3 by single or synergistic administration for 48 hours in vitro.</p><p><b>RESULTS</b>Gemcitabine above 10(-8) mol/L enhanced the growth suppression [suppression ratio > or = (50.8 +/- 4.2)%, P < 0.05] and apoptosis [apoptosis ratio > or = (22.9 +/- 2.3)%, P < 0.05] and down-regulation of the expression of cyclin D1 [expression ratio < or = (9.6 +/- 1.6)%, P < 0.01] induced by paclitaxel above 10(-7) mol/L in PC-3 cells. Gemcitabine changed the ratio of G2/M cell arrest induced by paclitaxel from (70.3 +/- 9.7)% to (38.2 +/- 4.2)%, and reversed the G2/M arrest partially (P < 0.01).</p><p><b>CONCLUSION</b>Paclitaxel and gemcitabine can enhance the growth suppression and apoptosis induced by paclitaxel in a synergistic way. They show great potential in the treatment of androgen-independent carcinoma of the prostate.</p>
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Humans , Male , Antimetabolites, Antineoplastic , Pharmacology , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Cell Line, Tumor , Deoxycytidine , Pharmacology , Dose-Response Relationship, Drug , Down-Regulation , Drug Synergism , Flow Cytometry , Paclitaxel , Pharmacology , Prostatic Neoplasms , PathologyABSTRACT
Objective To study the expression of MN mRNA in peripheral blood of renal cell carcinoma(RCC) patients and to evaluate its clinical significance. Methods Peripheral blood MN mRNA expression was detected by means of RT PCR 30 patients with renal cell carcinoma,including 16 with localized RCC and 14 with metastatic RCC,in 30 patients without renal tumors series as controts. Results With RT PCR, MN mRNA expressin was detected in 13 of 30 cases of RCC (43%) including 5 of 16 patients(31%) with localized RCC and 8 of 14 patients(57%) with metastatic RCC, none of the 30 control cases being positive. No correlation was noted between MN results and tumor stage. Conclusions RT PCR assay of MN is a highly sensitive technique for detecting circulating renal carcinoma cells in the peripheral blood, and it may be helpful in the diagnosis of micrometastasis.MN can be regarded as a cell specific relative protein of clear cell renal carcinoma.