Effect and Mechanism of RBM8A on Proliferation, Migration and Apoptosis of Human Endometrial Cancer HEC-1A Cells / 肿瘤防治研究

Objective To investigate the effect of silenced RBM8A gene on the biological behavior (proliferation, migration, and apoptosis) of human endometrial cancer HEC-1A cells and its possible mechanism. Methods The hairpin shRNA targeted by the RBM8A gene was designed, and the best shRNA silencing fragment was screened. The recombinant lentiviral interference vector carrying the target gene was constructed and used to infect HEC-1A cells. Cells with stable knockdown of RBM8A gene were screened by puromycin as the experimental group (shRBM8A), while the shRNA of nonsense sequence was designed as the control group (shControl). CCK-8 method was used to detect cell proliferation, and flow cytometry was used to detect cell apoptosis. Transwell assay was used to detect cell migration and invasion. Western blot was used to analyze the expression of apoptosis-related proteins and EMT signal transduction pathway related proteins. Results In comparison with the shControl group, after RBM8A knockdown, HEC-1A cell proliferation was reduced, apoptosis was increased, migration and invasion ability were significantly inhibited (P < 0.05), the expression of apoptosis-related proteins cleaved caspase 9 and caspase 3 increased, EMT-related protein E-cadherin expression increased, and Vimentin expression decreased. Conclusion RBM8A gene silencing can inhibit the proliferation, migration, and invasion and promote the apoptosis of endometrial cancer cells. The inhibition of EMT signal transduction pathway may be its mechanism.

Study on the Effect of Hydrophilic/Hydrophobic Nano-silica with Different Adding Amount on the Stability of Glycyrrhetinic Acid Lipo-emulsion / 中国药房

OBJECTIVE:To study the effect of hydrophilic/hydrophobic nano-silica with different adding amount on the stabili-ty of lipo-emulsion. METHODS:Glycyrrhetinic acid lipo-emulsion 4 mL was taken,respectively adding into 0.5%,1.0%,1.5%(m/m,the same below) hydrophilic nSiO2,and 0.4%,0.75,1.0% hydrophobic nSiO2,incubating 2 h in 30 ℃ water;the same batch of Glycyrrhetinic acid lipo-emulsion was treated as blank control. The forms were observed under electron microscopy after treatment,absorbance value was determined,the stability parameter (KE) was calculated according to the absorbance value,then the adding amount of nSiO2 was optimized,3 batches of preparations was prepared,and the verification test was conducted. RE-SULTS:The spherical structure was Glycyrrhetinic acid lipo-emulsion in the electron microscopy,the substance wrapping its sur-face white ring (fully wrapped) or semi-circular structure (not fully wrapped) was nSiO2. KE of hydrophilic nSiO2 were 4.66%, 5.01% and -2.08%,and KE of hydrophobic nSiO2 were 3.02%,4.51% and 7.24%. The optimized adding amount of hydrophilic nSiO2 was 0.2%,0.3% and 0.4%,hydrophobic nSiO2 was 0.1%,0.2% and 0.3%;KE were 6.19%,3.05%,7.84%,8.42%, 2.41%,2.93%,respectively. The optimal adding amount was 0.3% hydrophilic nSiO2 and 0.2% hydrophobic nSiO2;the 3 batches of preparation showed the optimum stability in its own adding amount. CONCLUSIONS:Both hydrophilic and hydrophobic nSiO2 can improve the stability of Glycyrrhetinic acid lipo-emulsion,and preferably 0.3%,0.2%.

Quality Evaluation and Index Exploration of the Glycyrrhetinic Acid Lipo-emul / 中国药房

OBJECTIVE:To provide reference indexs for the quality evaluation of Glycyrrhetinic acid lipo-emul. METHODS:Croy-TEM was used to detect the morphology of Glycyrrhetinic acid lipo-emul,laser nano-particle size analyzer was used to deter-mine the particle size,polydispersity index(PDI)and the zeta potential;UPLC was used to determine the drug loading of its ac-tive ingredient glycyrrhetinic acid;placing 10 d in 30 ℃,then stability was detected. RESULTS:Prepared Glycyrrhetinic acid li-po-emul was clear outline,structural integrity,roundlike and arranged closely;the mean particle size was(245.2±4.29)nm,PDI was (0.054 ± 0.01) and the mean zeta potential was (-6.25 ± 0.54) mV;the average drug loading of glycyrrhetinic acid was (1.25±0.09)mg/mL;the sample with mass concentration of 0.82 mg/mL showed good stability in within 10 d. CONCLUSIONS:Particle size,zeta potential,drug loading and stability can be used as the evaluation index of Glycyrrhetinic acid lipo-emul.

Effects of Genistein on The Expression of Estrogen Receptor ? and c-fos Gene in Aorta in Ovariectomized Rat / 中华高血压杂志

Objective To study the effects of genistein(GST) on the expression of estrogen receptor ?(ER?) and the expression of c-fos gene in the aorta of ovariectomized rats.Methods Forty female Wistar rats were randomly divided into four groups:sham-operated(control),ovariectomized(OVX),ovariectomized with 17?-E2 replacement(OVX+E2),ovariectomized with genistein replacement(OVX+GST) group.After 8 weeks' replacement therapy,the rats were sacrificed and the expression of ER? and c-fos in the aorta was studied by immunohistochemistry.Results Ovariectomy significantly decreased ER ? expression and increased c-fos expression in aorta;while replacement therapy,GST and E2,attenuated the effect of ovariectomy manifested by increasing ER expression and decreasing c-fos expression in aorta.Conclusion GST may modulate the expression of ER? in the arteries of ovariectomized rats and play a beneficial effects on cardiovascular system which was associated with decreases in expression of c-fos gene in aorta.

Preparation and Quality Standard of Bifonazole Hollow Effervescent Suppository / 中国药房

OBJECTIVE: To prepare bifonazole hollow effervescent suppository and establish its quality standard. METHODS: Bifonazole hollow effervescent suppository was prepared by hot-melting method, 38-type semisynthetic glyceride as the base material, NaHCO3 and citric acid as the effervescent agent, bifonazole as main ingredient. UV spectrophotometry was used for the content determination of bifonazole. RESULTS: The weight of prepared suppository was about 2.3 g and mean size of maximal foaming capacity were all above 8 mL. The linear range of bifonazole were 1.015～10.15 ?g?mL-1(r=0.999 9) with an average recovery of 99.74% (RSD=1.28%). CONCLUSION: The established preparation is simple, practical and up to the standard. The content determination method is precise and accurate.

Cardiovascular protective effects of soy isoflavone / 中国组织工程研究

OBJECTIVE: Estrogen has cardiovascular protective effects while its adverse effects restrain its application in the therapy and prevention of cardiovascular diseases. To find a more effective and safer estrogen replacement becomes a hotspot in cardiovascular pharmaceutical researches. This paper summarized the current research situation on the cardiovascular protective effects of phytoestrogen, soy isoflavone.DATA SOURCES: Relative articles between January 1993 and December 2001 were searched by computer on Medline with the searching words of "isoflavones, atherosclerosis, vasodilation" in English, the language limitation of the articles. Simultaneously, articles between January 1994 and February 2002 were searched by computer on Wangfang Database and Chinese Journal Full Text Database with the searching words of "isoflavones,artherosclerosis, vasodilation(Chinese charcters)" in Chinese, the language limitation of the articles.DATA SELECTION: Literatures with experiments including study group and control group were selected from the data through preliminary screening to eliminate obvious non-randomized experimental studies. The full texts of the residual literatures were searched afterwards for the further judgment of ranincluded in the study. Exclusive criteria: repetitive experimental studies.DATA EXTRACTION: A total of 31 randomized or non-randomized experimental articles regarding the cardiovascular protective effects of soy isoflavone were collected, of which 26 experiments were in accordance with the inclusive criteria and the rest 5 articles were exclude due to repetition of same study.DATA SYNTHESIS: Twenty-six experiments including clinical experiments and animal experiments, which employed in vivo or in vitro two experimental methods after the application of soy isoflavone to observe and evaluate its cardiovascular protective effects. The above two methods had its own merits and shortcomings, of which in vitro experiment was a more common method for the observation.CONCLUSION: There is no adequate evidence that can prove the definite effects of soy isoflavone on the prevention and therapy of cardiovascular diseases in menopausal women, as well as its side effects. Researchers should do more researches on how to master the appropriate dose to make isoflavone reach effective blood concentration, and how to make the therapeutic effects become more beneficial.

Effect of retinoid kappa receptor alpha (RXRalpha) transfection on the proliferation and phenotype of rat hepatic stellate cells in vitro / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To study the effect of retinoid kappa receptor alpha (RXRalpha) transfection plus treatment with the RXRalpha ligand, 9-cis-RA, on the proliferation and phenotype of platelet-derived growth factor (PDGF) activated hepatic stellate cells (HSCs).</p><p><b>METHODS</b>PDGF activated rat hepatic stellate cells were transfected with eukaryotic expression vector pcDNA3.1- human RXRalpha, and confirmed by Western blot. Proliferation of transfected HSC was assayed by bromodeoxyuridine (BrdU) incorporation as well as MTT, and the phenotype (alpha-smooth muscle actin, desmin) was observed by immunocytochemistry with image analysis.</p><p><b>RESULTS</b>Transfection of the RXRalpha gene and treatment with ligand 9-cis-RA of PDGF-activated HSCs extended the increased expression of RXRalpha protein for at least 168 hours. Cell proliferation and expressions of alpha- smooth muscle actin (alpha-SMA) and desmin were blocked, compared with groups of sham-transfected, PDGF-activated, no transfection, no ligand treatment, and irrelevant ligand treated HSCs.</p><p><b>CONCLUSION</b>Transfection with the RXRalpha gene followed by 9-cis-RA ligand treatment will inhibit the proliferation and reverse the phenotype of activated HSC.</p>

Urinary type IV collagen: a specific indicator of incipient diabetic nephropathy / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To determine whether urinary type IV collagen can serve as an indicator specific for diabetic nephropathy.</p><p><b>METHODS</b>Using a novel sandwich ABC-ELISA to measure type IV collagen directly, the 24-hour urinary type IV collagen excretion rate was determined in 120 diabetic patients and some groups of controls. Urinary albumin determinations were made with a RIA kit at the same time. A total of 13 diabetic patients with microalbuminuria underwent percutaneous renal biopsy for definitive diagnosis of diabetic nephropathy. Type IV collagen and TGF-beta 1 immunoreactivities were detected with ABC methods in renal biopsies.</p><p><b>RESULTS</b>Urinary type IV collagen excretion was significantly increased in diabetic patients with microalbuminuria, especially those with albumin excretion above 200 mg/24 h. By comparison, collagen excretion was equivalent to that in healthy controls when measured in diabetics with normalbuminuria and in patients with primary glomerular disease, primary hypertension, or coronary heart disease. Urinary type IV collagen excretion in diabetics was negatively correlated with creatinine clearance. In renal biopsies from subjects with elevated collagen excretion, the glomeruli showed pathological changes typical of diabetic nephropathy. Also, excessive type IV collagen and TGF-beta 1 immunoreactivity were detected in the glomeruli, Bowman's capsule and interstitium.</p><p><b>CONCLUSIONS</b>Excretion of type IV collagen, possibly reflecting increased production or decreased degradation of this protein, may be a clinically useful indicator of incipient diabetic nephropathy.</p>

The expressions of TGF-beta1 and Smad 2 mRNA on diseased glomeruli and their significance in the development of glomerulosclerosis / 中华病理学杂志

<p><b>OBJECTIVE</b>To explore the expressions of transforming growth factor-beta1 (TGF-beta1) and its signaling transduction molecule Smad 2 and their significance in the development of glomerulosclerosis.</p><p><b>METHODS</b>Using in situ hybridization and immunohistochemistry to detect Smad 2 mRNA expression and TGF-beta1, collagen IV, fibronectin expression in renal biopsies from 61 cases with a spectrum of glomerulonephritis including IgA nephropathy (40 cases), membranous glomerulonephritis (10 cases) and sclerosing glomerulonephritis (11 cases), compared with 11 cases of glomerular mild lesion with image analysis system.</p><p><b>RESULTS</b>With the exception of Smad 2 mRNA expression in mild type IgA nephropathy, all other types of diseased glomeruli showed increased expression of both TGF-beta1 and Smad 2 mRNA when compared with the 11 cases of mild glomerular lesions. The expressions of glomerular TGF-beta1 and Smad 2 mRNA positively correlated with collagen IV and fibronectin deposition in the glomeruli.</p><p><b>CONCLUSIONS</b>TGF-beta1 and Smad 2 may be involved in the excessive deposition of glomerular extracellular matrix and play an important role in the development of glomerulosclerosis.</p>

The localization of adrenomedullin in rat kidney tissue and its inhibitory effect on the growth of cultured rat mesangial cells / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To observe the localization of adrenomedullin (AM) in rat kidney tissue and its inhibitory effect on the growth of cultured rat mesangial cells (MsC).</p><p><b>METHODS</b>A monoclonal antibody against AM developed by our laboratory was used to detect the localization of AM protein in rat kidney tissue by avidin-biotin complex immunohistochemistry. The expressions of AM and its receptor CRLR mRNA on cultured glomerular epithelial cells (GEC) and MsC were investigated by Northern blot assay, and the possible effect of AM secreted by GEC on MsC proliferation was observed using [3H]thymidine incorporation as an index.</p><p><b>RESULTS</b>A specific monoclonal antibody against AM was succesfully developed. AM was immunohistochemically localized mainly in glomeruli (GEC and endothelial cells), some cortical proximal tubules, medullary collecting duct cells, interstitial cells, vascular smooth muscle cells and endothelial cells. Northern blot assay showed that AM mRNA was expressed only on cultured GEC, but not on MsC, however, AM receptor CRLR mRNA was only expressed on MsC. GEC conditioned medium containing AM can inhibit MsC growth and AM receptor blocker CGRP8-37 may partially decreased this inhibitory effect.</p><p><b>CONCLUSION</b>AM produced by GEC inhibits the proliferation of MsC, which suggests that AM as an important regulator is involved in glomerular normal physiological functions and pathologic processes.</p>

Significance of u-PA and PAI-1 Expression on Diseased Glomeruli / 复旦学报（医学版）

Purpose To investigate the significance of u-PA and PAI-1 expression in various types ofglomerulonephritis. Methods The expression level of u-PA, PAI-1, type-Ⅳ collagen and PCNA positivenuclei in 120 cases of renal biopsies from patients with various types of glomerulonephritis were detected byimmunohistochemical method. Both the staining intensity of u-PA and PAI-1 in glomeruli were quantitativelyanalyzed by image analysis. Results The expression intensity of u-PA and PAI-1 was different in varoustypes of glomerulonephritis, which were significantly higher than that of the minor lesion group. Meanwhile,the intensity of PAI-1 stain was significantly higher than that of u-PA in various types of glomerulonephritis( P ＜ 0.05). The increase of u-PA expression was closely related to increase of type- Ⅳ collagen synthesis andhypercellularity in glomeruli(r = 0. 761 and 0. 811, P＜ 0.05), while the expression of PAI-1 was closelyrelated to the increase of Col-Ⅳ synthesis other than the cell proliferation in the glomeruli. ConclusionsThe over expression of u-PA and PAI-1 may play an important role in contributing to the pathogenesis anddevelopment of glomerulonephritis.

A Study of Apoptosis in Renal Tissue of Diffuse Proliferative Lupus Nephritis / 复旦学报（医学版）

Purpose To evaluate apoptosis in renal tissue of diffuse proliferative lupus nephritis and therelationship between the existence of apoptosis cells in renal tissue and histopathological or clinical changes.Methods Apoptosis was detected by in situ nick-end labeling techniques (TUNEL) in renal biopsies from 25patients with type Ⅳ LN, 12 patients with IgAN, 4 patients with MsPGN, and 3 patients with APSGN. Normalrenal tissue obtained at nephrectorny for hypemephroma in 4 adults was used as control. In addition, proliferatingcells were identified by proliferating cell nuclear antigen(PCNA) in these patients. Results Compared to otherproliferative glomerulonephritis and control,the patients with lupus nephritis had less apoptosis cells, higher ratio ofPCNA+ cells/TdT+ cells/(P/T) in renal tissues;Ratio of P/T in glomeruli and tubulointerstitium correlated withthe chronicity index, r=0. 498 3(P = 0. 013 2), r = 0. 839 9(P＜ 0.001 ), r = 0. 661 4(P = 0. 003 3),respectively. Ratio of P/T in glomerulus and tubule had positive correlation with 24 hour urinary protein, r =0.855 4(P＜0.001),r=0.713 4(P=0. 001); negative correlation with Ccr, r = - 0. 488 0(P =0. 013 3)and r = - 0. 722 9(P = 0. 001), which in tubules positively correlated with Scr, r = 0. 410 7 (P = 0.041 4 ).Conclusions Apoptosis is insufficient in proliferative lupus nephritis. Intense proliferation without followingincrease in apoptosis may be related to chronic progressive renal histopatholcgical changes.

Effect of erythromycin on interdigestive and postprandial gastrointestinal contractile activity and its mechanism in dogs / 中国药理学通报

AIM　To observe the effect of intravenous injection of erythromycin (EM) on interdigestive migrating motor complex (MMC) and postprandial gastrointestinal contraction in conscious dogs , and to study its possible mechanism. METHODS　Gastrointestinal contractile activity was recorded using low compliance capillary water perfusion manometric system. EM was administered intravenously during phaseⅠ and after meal, and blood samples were drawn for measuring plasma motilin concentrations. RESULTS　①Plasma motilin levels showed cyclical fluctuations in different phases of MMC, and plasma motilin reached peak during phaseⅢ and lowest during phaseⅠ.②EM induced phaseⅢ-like contractions in the antrum and duodenum, which was not accompanied by a peak in plasma motilin level. The optimum dose of EM for resulting in a premature phaseⅢ with the same characteristics as the spontaneously occurring phaseⅢ was established to be 0.5 mg*kg-1. The dose of 10 mg*kg-1 EM induced gastrointestinal continuous contractions and duodeno-gastric retrograde peristalsis which was associated with retching and vomiting. ③Atropine obviously inhibited EM-induced phaseⅢ activity in the antrum and duodenum. ④EM powerfully enhanced postprandial gastrointestinal contractile activity. CONCLUSIONS The results suggests that EM is a potent prokinetic agent. The mechanism is not related to the release of motilin, but may be mediated partially by cholinergic pathway.

Effect of Heparin on the Cell Proliferation and u-PA/PAI-1 Expression on Glomeruli in Rat Anti-thy1 Glomerulonephritis / 复旦学报（医学版）

Purpose　To investigate the significance of u-PA and PAI-1 expression on the glomeruli,and the effect of heparin on their expressions in rat anti-thy1 glomerulonephritis.　Methods　We analyzed the cell proliferation and the expression of u-PA/PAI-1 on the glomeruli by immunohistochemistry and quantitative analysis of immunostaining.　Results　The cell proliferation of the glomeruli decreased significantly at 7 th,14 th,21 st day after heparin treatment in comparison to the glomerulonephritic group(P<0.05 or 0.01).The expression of u-PA and PAI-1 on the glomeruli in glomerulonephritic and heparin-treated groups was higher than that in the control group.At 3 rd,7 th,14 th,21 st day,the glomerular hypercellularity in the glomerulonephritic group was closely related to the increased expression of u-PA and PAI-1(P<0.05 or 0.01).At 3 rd,7 th day,the decreased cell proliferation of the glomeruli in heparin-treated group had close relationship with the decreased expression of PAI-1(P<0.05).　Conclusions　In rat anti-thy 1 glomerulonephritis model,the expression of u-PA and PAI-1 increased with glomerular hypercellularity;heparin treatment can decrease the extent of glomerular hypercellularity in rat anti-thy 1 glomerulonephritis.The treatment function of heparin might be related with the inhibitory effect of PAI-1 expression on the glomeruli.

Expression of ANGPTL3 in children with primary nephrotic syndrome / 中华肾脏病杂志

Objective To examine the expression of angiopoietin-like protein(ANGPTL)3 in kidneys from children with primary nephrotic syndrome. Methods Immunohistochemistry for ANGPTL3 was performed in kidney biopsies from patients with nephrotic syndrome or hematuria, including MCD (n=31), MN(n=6), FSGS (n=6), TBMN (n=10), IgA nephropathy (IgAN) with mesangial proliferation (n=16). Normal renal tissue of 2 cases with nephrectomy for tumor were used as control. According to the episode, four groups were divided ("12 months"). The expression was quantitatively examined with IMS color image analysis system, using positive index (PI) as sediment degree of ANGPTL3 in glomeruli or tubules. Immunofluorescence for ANGPTL3 co-labeling with WT1 and perlecan was applied to show the distribution of ANGPTL3. Results (1) The PI levels of ANGPTL3 in glomeruli of MCD(7.49?1.96) and MN (6.27?0.98) were significantly higher than those of TBMN (0.02?0.001), FSGS (3.14?0.49) or normal control(0.02?0.001) respectively (all P

Expression of apoiipoprotein H in childhood primary nephrotic syndrome / 中华肾脏病杂志

Objective To study the expression of apolipoprotein H (ApoH) in childhood primary nephrotic syndrome (PNS) and to discuss its role in PNS. Methods Immunohistochemistry staining and real-time quantitative polymerase chain reaction(RT-PCR) were performed to evaluate the expression of ApoH in renal tissues of 78 patients with PNS and 14 cases of normal controls. Serum albumin, serum lipid, proteinuria and urinary retinol binding protein (RBP) were tested before renal biopsy in all patients. Tubulointerstitial lesions were investigated. Results (l)There was positive expression of ApoH in renal tissues of PNS patients and normal controls,mainly in the proximal tubules by immunohistochemistry staining. ApoH mRNA was also shown in all renal tissues by RT-PCR. ApoH protein expression was positively correlaed with its mRNA expression(r=0.264, P 0.05) whereas these data displayed no significant difference between two groups. Above expression in mesangial proliferative glomerulonephritis (MsPGN) and focal segmental glomersclerosis (FSGS) down-regulated significantly (3.30?0.28,2.82?0.36, and 10.13?3.09,10.12?1.02, respectively), as compared to those in MCNS,MN and the controls, P

Effect of erythromycin on interdigestive and postprandial gastrointestinal contractile activity and its mechanism in dogs / 中国药理学通报

AIM To observe the effect of intra- venous injection of erythromycin (EM) on interdigestive migrating motor complex (MMC) and postprandial gastrointestinal contraction in conscious dogs, and to study its possible mechanism. METHODS Gastrointestinal contractile activity was recorded using low compliance capillary water per fusion manometric system. EM was administered intravenously during phase I and after meal, and blood samples were drawn for measuring plasma motilin concentra- tions. RESULTS ①Plasma motilin levels showed cyclical fluctuations in different phases of MMC, and plasma motilin reached peak during phaseⅢ and lowest during phase Ｉ. ②EM induced phase Ⅲ -like contractions in the antrum and duodenum, which was not accompanied by a peak in plasma motilin level. The optimum dose of EM for resulting in a premature phaseⅢ with the same characteristics as the spontaneously occurring phaseⅢ was established to be 0. 5 mg.kg-1. The dose of 10 mg.kg-1 EM induced gas- trointestinal continuous contractions and duodeno-gas-tric retrograde peristalsis which was associated with retching and vomiting. ③Atropine obviously inhibited EM-induced phase Ⅲ activity in the antrum and duodenum. GEM powerfully enhanced postprandial gastrointestinal contractile activity. CONCLUSIONS The results suggests that EM is a potent prokinetic agent. The mechanism is not related to the release of motilin, but may be mediated partially by cholinergic pathway.