Contrastive analysis of corneal horizontal diameter measurement by five instruments / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To analyze the correlation and consistency of five corneal diameter measurements.<p>METHODS: Totally 25 cases(50 eyes)who underwent ICL implantation in West China Hospital. The preoperative horizontal corneal diameter was measured using measuring caliper, SIRIUS anterior eye assay system, IOL Master500, anterior segment OCT and UBM. <p>RESULTS: The mean WTW distances were 11.54±0.30 mm as obtained with measuring caliper, 11.77±0.33mm with SIRIUS anterior eye assay system, 11.98±0.33mm with IOL Master500, 11.63±0.35mm with anterior segment OCT and 11.53±0.34mm with UBM. No statistical difference was found between measuring caliper and UBM, measuring caliper and anterior segment OCT, UBM and anterior segment OCT. The linear correlation analysis found significant correlation between the measurements of the five measurements. The Bland-Altman analysis for the measuring caliper and SIRIUS, measuring caliper and UBM, measuring caliper and anterior segment OCT found that the absolute values of 95% LOA upper and lower limits were less than 0.5mm.<p>CONCLUSION: The results of the four kinds of corneal horizontal diameter measurements can be interchanged including SIRIUS, UBM, anterior segment OCT and measuring caliper. IOL Master500 results are the largest, that cannot be used as a diagnostic basis for measuring the size of corneal horizontal diameter. The results of other measurement equipment should be combined with clinical practice.

Consistency of two eye anterior segment analysis systems measuring wave-front aberration before and after refractive surgery / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To study the consistency of pre-operative and postoperative wave-front aberration between SCHWIND and SIRIUS eye anterior segment analysis system. <p>METHODS: Prospective control study. A total of 360 eyes of 180 patients with refractive errors in West China Hospital of Sichuan University were selected. The data, including the degree and the direction of the flat axis and the steep axis, Kappa angle and a variety of high order wave-front aberrations, were measured by SCHWIND and SIRIUS anterior segment analyzers. The average value of each parameter was taken 5 times, and paired sample <i>t</i> test was used and the data was analyzed by SPSS22.0 software. <p>RESULTS: Flat axis degree and direction, steep axis degree and direction, coma, and trefoil were measured preoperatively and 1mo postoperatively by SCHWIND and by SIRIUS eye anterior segment analysis systems, and there were no statistically significant differences between the two machines(<i>P</i>>0.05). Kappa angle was measured 1mo postoperatively and there was no statistically significant differences(<i>P</i>>0.05). There was statistically significant difference(<i>P</i><0.01)preoperatively between two machines of high-order aberrations and spherical aberration(SpAb); values measured by SCHWIND were higher than those measured by SIRIUS. There was no statistically significant difference(<i>P</i>>0.05)between two machines of high-order aberrations or SpAb in 1mo postoperatively. <p>CONCLUSION: SCHWIND and SIRIUS anterior segment eye analysis system have a good consistency of wave-front aberration measurement before and after refractive surgery.

Comparison of corneal thickness measured by CASIA SS-1000 OCT and Sirius anterior eye assay system / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To compare CASIA SS-1000 and Sirius OCT sweep anterior segment analyzer instrument for measuring normal corneal vertex thickness (CCT) and the thinnest corneal thickness (TCT) results the difference,correlation and consistency,and provide a theoretical basis for clinical application.METHODS:This was a prospective study.A total of 34 normal subjects were collected.The subjects were measured by the same skilled operator.The SS-1000 OCT was first used,and then the corneal thickness was measured repeatedly by Sirius anterior segment analyzer.Paired t test and Bland-Altman were used to evaluate the consistency of corneal apex and corneal thinnest point between SS-1000 OCT and Sirius anterior segment analyzer.RESULTS:The mean corneal apex measured by SS-1000 OCT and Sirius corneal topography were 517.62± 25.29μm and 518.47±27.23μm CCT,respectively.The thinnest points of SS-1000 OCT and Sirius anterior segment analyzer CCT were 513.53±25.06μ m and 515.32± 26.69μm,respectively.Paired t test showed that the difference on corneal thickness of vertex was not statistically significant (P＞0.05),but the thinnest corneal thickness was statistically significant (P＜ 0.05).Pearson analysis of the two devices,the correlation is 0.969,0.965.The results of 95％ consistency limiting analysis on the corneal vertex thickness by Bland-Altman was (-14.22μm,12.52μm),that of the thinnest corneal thickness was (-15.61μm,12.03μm),4％ (3/68) was out of the 95％ consistency limiting,but the thinnest corneal thickness was of a little larger differences.CONCLUSION:SS-1000 OCT measurement of CCT and Sirius anterior segment analyzer is highly consistent,in clinical work can be considered alternative,but the thinnest point of the cornea can not be replaced each other.

Validation and Evaluation of Diagnostic Efficiency of Genes Associated with Colorectal Cancer with Hyperglycemia / 中国医学科学院学报

Objective To verify the expressions of genes associated with colorectal cancer with hyperglycemia and evaluate their diagnostic values.Methods Tumor tissues,distal normal intestinal mucosa,and peripheral blood samples were harvested from 109 colorectal cancer patients and peripheral blood samples from 30 diabetes patients and 30 healthy volunteers. The mRNA expressions of glucose regulated protein 78 (GRP78),NADPH oxidase-1 (NOX1),carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5),heat shock protein 60 (HSP60),and histone deacetylase 1(HDAC1) were detected by real-time quantitative polymerase chain reaction. The correlation between the gene expressions and clinicopathological parameters in colorectal cancer patients were analyzed using Pearson's correlation analysis. Diagnostic test accuracy evaluation was used to calculate the sensitivity,specificity,accuracy,predictability,Youden index,and likelihood ratio of serum gene expressions in colorectal cancer patients,and the receiver operating characteristic (ROC) curves were drawn. The area under the ROC curve was calculated to evaluate the diagnostic efficiency of the combined detection of multiple genes.Results The mRNA levels of GRP78 (P=0.001),NOX1 (P=0.022),CEACAM5 (P=0.000),HSP60 (P=0.044),and HDAC1 (P=0.047) were positively correlated with the fasting blood glucose level. The mRNA expressions of NOX1 (P=0.000,P=0.008) and HDAC1 (P=0.000,P=0.037) in tissues and serum were significantly higher in colorectal cancer patients than in patients with normal blood glucose levels. The NOX1 mRNA expression was positively correlated with the diameter of colorectal cancer (P=0.013),and the HDAC1 mRNA expression was significantly correlated with the tumor site (P=0.049),depth of primary tumor invasion (P=0.025),and TNM stage (P=0.042). The areas under the ROC curves of NOX1,CEACAM5,and HDAC1 were 0.931,0.852,and 0.860 respectively (all P=0.000). The specificity,accuracy,and negative predictive value of NOX1,HDAC1 mRNA expression in colorectal cancer patients with hyperglycemia were all above 90%. The diagnostic sensitivity and specificity of the combined detection of NOX1,CEACAM5,and HDAC1 were 98.82% and 99.93%,respectively.Conclusion Combined detection of genes associated with colorectal cancer accompanied by hyperglycemia can improve the diagnostic efficiency of early screening.

A comparison study on influencing factors of two different successful aging models / 预防医学

Objective To explore the differences between two successful aging models including three -dimensional model and four -dimensional model by comparing the influencing factors.Methods A total of 230 elderly people in sanatorium were selected by random sampling method.Then questionnaires and scales were conducted and the influencing factors of the two theoretical models were tested by binary logical regression analysis.Results 86 elderly people (37.39%)were qualified as successful aging by three -dimensional model (chronic illness,physical and psychological functions,social functions)while 51 elderly people (22.17%)were qualified by four -dimensional model (chronic illness,physical and psychological functions,social function,subjective well -being).When using three -dimensional model,two factors reached significant level on successful aging phenomenon:age and education background,which could explain 69.10% of the total variance.When using four -dimensional model,three factors finally showed the significance:age,psychological resilience and subjective support,which could explain 79.12% of total variance.Conclusion The qualification of successful aging status can be largely influenced by evaluation models.

The mechanisms of inhibitory effect of adenovirus-mediated wild-type PTEN gene on the proliferation in activated hepatic stellate cells in vitro / 中华肝脏病杂志

<p><b>OBJECTIVE</b>Using an adenoviral vector, the wild-type PTEN gene was transduced into activated hepatic stellate cell (HSC) cultured in vitro and cell cycle markers and were detect. Thereby, the potential mechanisms of inhibitory effect of the wild-type PTEN overexpression on the proliferation in activated HSC was investigated.</p><p><b>METHODS</b>The wild type PTEN gene was transduced into activated HSC (HSC-T6 ) cultured in vitro mediated by adenoviral vector. PTEN expression in HSC was measured by Western blot and Real-time fluorescent quantitation PCR. Flow cytometry (FCM) was then used to detect cell cycle phase of activated HSC. And the expressions of cyclinD1 and cyclin dependent kinase 4 (CDK4) in HSC were determined by Western blot.</p><p><b>RESULTS</b>The data showed that exogenous wild type PTEN gene was successfully transduced and expressed in activated HSC cultured in vitro. The over-expression of wild type PTEN resulted in the increased number of HSC at G0/G1 phase ( P less than 0.01), and the number of HSC at S phase and G2/M phase were decreased significantly, P less than 0.01. Furthermore, there were decreased cyclinD1 and CDK4 expression in HSC infected with Ad-PTEN, P less than 0.01.</p><p><b>CONCLUSION</b>The over-expression of wild type PTEN inhibit transition of activated HSC in vitro from G1 to S phase and arrest cell cycle of them at G0/G1 phase via the down-regulated expressions of cyclinD1 and CDK4, and then inhibit HSC proliferation.</p>

Aldosterone stimulates hepatic stellate cells contraction via Ca2+-independent pathways / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the mechanisms of Aldosterone stimulating hepatic stellate cells(HSCs) contraction via Ca2+-independent pathways.</p><p><b>METHODS</b>HSC-T6 cell line was pre-disposed with Aldo 10mumol/L. The cell contraction was detected by silicone-rubber-membrane cultivation directly. The concentration variation of intracellular free calcium in rat HSC was observed by laser confocal microscopy. Besides, HSC-T6 cell line was under pre-disposal treatment with the blocking agents of Aldo receptor -antisterone, protein kinase C (PKC) special blocking agent-Stauro, Rho kinase blocking agent-Y27632 and MLCK special blocking agent-ML-7 respectively prior to stimulation with aldosterone. RT-PCR was used to detect the expression of Rock2, RhoAGTP and RhoGEF in Ca2+- independent pathways mediated by Rho-kinase.</p><p><b>RESULTS</b>Aldo could induce HSCs contraction. The concentration of intracellular free calcium in rat HSCs had no change after pre-disposal treatment with Aldo. The mRNA expressions of Rock2, RhoAGTP and RhoGEF increased significantly after treatment with Aldo (0.770+/-0.049, 0.960+/-0.096, 0.180+/-0.006, P is less than 0.01).When inhibited with antisterone, the mRNA expressions of the three elements were (0.440+/-0.166, 0.370+/-0.180 and 0.050+/-0.001, P is less than 0.01), lower than that of Aldo group, but higher in ML-7+Stauro + Aldo groups (0.940+/-0.066, 1.330+/-0.192 and 0.160+/-0.007, P is less than 0.05) as compared to the control group (0.140+/-0.023, 0.540+/-0.111 and 0.110+/-0.012). In the Y27632 + ML-7 + Stauro+Aldo group, the mRNA expression of RhoGEF (0.290+/-0.004, P is less than 0.01)was higher than that of the ML-7 + Stauro + Aldo group (0.160+/-0.007).</p><p><b>CONCLUSION</b>Aldo could induce HSCs contraction via Ca2+-independent pathways and Rho-Rock pathway involved in the process.</p>

The role of extracellular signal-regulated kinase in induction of apoptosis with salvia miltiorrhiza monomer IH764-3 in hepatic stellate cells / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the effect of Salvia miltiorrhiza monomer IH764-3 on apoptosis in hydrogen peroxide (H2O2)-stimulated hepatic stellate cells (HSCs).</p><p><b>METHODS</b>HSCs were cultured in medium with different IH764-3 doses (10 mg/L, 20 mg/L, 30 mg/L, 40 mg/L) and without IH764-3. Direct cell count, 3H-thymidine incorporation, Annexin-V/Propidium Iodide double-labeled flow cytometry, TUNEL and transmission electron microscopy were employed to estimate the influence of IH764-3 on proliferation and apoptosis of HSCs. The expression of extracellular signal-regulated kinase 1 (ERK1) mRNA and protein in HSCs were detected using RT-PCR and Western blot respectively.</p><p><b>RESULTS</b>It was showed that H2O2 could promote HSC proliferation. In contrast, IH764-3 at concentrations of 10 mg/L, 20 mg/L, 30 mg/L and 40 mg/L inhibited its proliferation. The inhibition rates were 7.13%, 28.36%, 53.80% and 73.10% (P < 0.01). And the inhibition rates of IH764-3 at concentrations of 30 mg/L at 12 h, 24 h and 48 h were 22.24%, 40.51% and 61.65%. Furthermore, IH764-3 could also induce the HSC apoptosis in dose-dependent an dtime-dependent manners (P < 0.01). In addition, after exposed of HSCs to IH764-3 for 24 h, ERK production decreased and ERK1 mRNA was down-regulated earlier about 2 h after exposure to IH764-3.</p><p><b>CONCLUSION</b>IH764-3 may inhibit the proliferation and induce apoptosis of HSCs in both dose-dependent and time-dependent manners, which may be related to down-regulation of ERK expression.</p>

Application of neoadjuvant chemotherapy for operable breast cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the clinical value of neoadjuvant chemotherapy in the treatment of operable breast cancer.</p><p><b>METHODS</b>A total of 120 patients with pathologically proven operable breast cancer were randomized into two groups to receive 2-4 cycles of neoadjuvant chemotherapy with docetaxel combined with pirarubicin (TThp) or docetaxel combined with epirubicin (TE). Operations were performed two weeks after the neoadjuvant chemotherapy. The short-term therapeutic effect, toxic reaction and the impact of neoadjuvant chemotherapy on the choice of surgical approaches were evaluated.</p><p><b>RESULTS</b>The total effective rate for the primary sites was 87.2% in these patients, and the rate of breast conservation significantly increased from 12.7% to 41.8% (P<0.05), with a tumor resection rate of 97.2%. The major adverse effects of the therapy included leukopenia, nausea, vomiting and alopecia.</p><p><b>CONCLUSION</b>Neoadjuvant chemotherapy can enhance the breast conservation rate, lower the clinical staging of the tumors and minimize the surgery area to improve the postoperative quality of life of the patients.</p>

FAK-related non-kinase plasmid transfection inhibited hepatic stellate cells proliferation / 中国应用生理学杂志

<p><b>AIM</b>To observe the effect of FAK-related non-Kinase (FRNK) plasmid on hepatic stellate cell (HSC) proliferation stimulated by fibronectin (FN).</p><p><b>METHODS</b>FRNK plasmid was transfected into HSC with transient liposomal transfection. The proteins of FRNK, FAK and p-FAK(Tyr397) were assayed by Western blotting analysis. The proliferation of HSC was evaluated by improved MTT assay, and cell cycle pattern was determined by flow cytometry (FCM).</p><p><b>RESULTS</b>(1) The expression of FRNK protein increased after FRNK transfected HSC, and it was at 48 h that the expression of FRNK protein was the highest (P < 0.01). The protein level of FAK was no significant difference between before FRNK plasmid transfection and after transfection (P > 0.05). The expression of p-FAK(Tyr397) protein was down-regulated after FRNK had been transfected in HSC, (P < 0.01). (2) The HSC proliferation inhibition rates at 12 h, 24 h and 48 h after FRNK transfection were 20.07%, 26.16%, 29.77%, respectively (P < 0.01). (3) Compared with the non-FRNK plasmid group, the FRNK-transfected HSCs almost arrested in G0/G1 phase (71.4 +/- 2.81 vs 48.9 +/- 1.66, P < 0.01).</p><p><b>CONCLUSION</b>After FRNK were transfected successfully in HSCs using lipofectamine, the phosphorylation of FAK was inhibited. The HSC proliferation was restrained in a time-dependent manner and the HSC was arrested in G0/G1 phase.</p>

The influence of down-regulation of focal adhesion kinase by RNA interference on the adhesion and migration of rat hepatic stellate cells in vitro / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the role of focal adhesion kinase (FAK) in adhesion and migration of hepatic stellate cells (HSC).</p><p><b>METHODS</b>Two recombinant plasmids expressing short hairpin RNAs (shRNAs) targeting FAK were constructed and one plasmid substantially suppressing FAK expression in HSC was selected. Real-time PCR and Western blot were used to detect the knockdown effects of FAK gene. After 48-hour treatment with FAK shRNA, toluidine blue colorimetric assay was used to detect the cell adhesion. Wound-healing assay and improved Boyden double-chamber were used to detect the cell migration induced by FN.</p><p><b>RESULTS</b>The recombinant plasmid expressing FAK shRNA was successfully constructed and transfected into HSC. Compared with the controls, the expression of FAK mRNA and protein in HSC treated with FAK shRNA was markedly down-regulated by 76.82% and 72.53%, respectively. The expression of p-FAK (Tyr397) protein was also decreased by 62.71% 48 h posttransfection. The adhesion of HSC was inhibited by 58.69% at 48 h after shRNA transfection. FAK gene silencing could also dramatically inhibit FN-stimulated HSC migration, and the cell migration distance and the cell number of crossing membrane were decreased by 58.27% and 83.70%, respectively.</p><p><b>CONCLUSIONS</b>FAK gene silencing suppresses adhesion and migration of HSC, and FAK may be a potential target for novel anti-fibrosis therapies.</p>

Dynamic changes of alpha-AR, beta1-AR and beta2-AR expression during hepatic fibrogenesis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the dynamic changes of a-AR, b1-AR and b2-AR expression in hepatic fibrosis.</p><p><b>METHODS</b>Rat hepatic fibrosis model was established by bile duct ligation (BDL). HE and Masson staining were used to determine hepatic fibrosis levels. Immunohistochemistry was applied to detect alpha -smooth muscle actin (alpha -SMA), a marker of hepatic stellate cell (HSC) activation; Western blot and real-time RT-PCR were used to measure the dynamic changes of alpha -AR, beta(1)-AR, beta(2)-AR expression on protein and mRNA levels, respectively, during the development of hepatic fibrosis.</p><p><b>RESULTS</b>(1) HE and Masson trichrome staining showed that the liver fibrosis models were established successfully. (2) At 1, 2, 3, 4 wk after BDL, alpha -SMA positive area density of the model group (10.58% +/- 1.75%, 24.14% +/- 2.02%, 29.74% +/- 2.59%, 34.28% +/- 2.01%) was significantly higher than that of the sham operation group (4.12% +/- 1.51%), P less than 0.01. (3) The expression of alpha -AR, beta(1)-AR, beta(2)-AR protein and mRNA was increased with the development of the hepatic fibrosis (P less than 0.05). (4) alpha -SMA expression was positively associated with alpha -AR, beta(1)-AR, beta(2)-AR, r values were 0.564, 0.753 and 0.606, respectively.</p><p><b>CONCLUSION</b>The expression of alpha -SMA is increased dramatically during the fibrosis, and is positively associated with the expression of alpha -AR, beta(1)-AR and beta(2)-AR.</p>

Changyanqing decoction produces anti-inflammatory effect by inhibiting the activation of nuclear factor-kappaB / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the changes in intestinal mucosal permeation in rats with methotrexate-induced small intestinal damage and investigate the protective effects of Changyanqing decoction.</p><p><b>METHODS</b>Rat enteritis model was established by methotrexate (MTX) and sodium chloride. The rats were randomly divided into normal control group, model group, N-acetylcysteine (NAC) group and Changyanqing decoction group, and Changyanqing decoction (100 mg/kg) or saline was administered daily in the corresponding groups by gastric irrigation for 6 days. The disease activity index (DAI), colonic mucosal damage index (CMDI) and histological score (HS) of the rats were observed and evaluated. The levels of mRNA expressions of TNF-alpha and IL-1beta were detected by semi-quantitative RT-PCR. The expression of IL-10 was detected by enzyme linked immunosorbent assay, and IkappaB expression was determined with Western blotting.</p><p><b>RESULTS</b>Compared with the normal control group, the model group showed significantly increased DAI, CMDI and HS. The DAI, CMDI, and HS in rats treated with Changyanqing decoction were significantly decreased in comparison with those in the model group (P<0.01). The expressions of TNF-alpha and IL-1beta were significantly higher in MTX-treated group than in the control group. The expression of TNF-alpha and IL-1beta mRNA in the Changyanqing group and NAC group were significantly lower, but IL-10 significantly higher than those of the MTX group. In MTX group, obvious NF-kappaB activation was observed, whose expression was significantly stronger in the cell nuclei, and the IkappaB in the cytoplasm was markedly degraded.</p><p><b>CONCLUSION</b>Changyanqing decoction offers protection on intestinal mucosa by inhibiting NF-kappaB activation to reduce TNF-alpha and IL-1beta mRNA expressions and increase IL-10 expression.</p>

The role of FAK-ERK signal transduction pathway in apoptosis of hepatic stellate cell / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the effects of FAK-related non-kinase (FRNK) on the apoptosis of hepatic stellate cells (HSC) in vitro and on the extracellular signal-regulated kinase (ERK) signal transduction pathway.</p><p><b>METHODS</b>HSC were stimulated by fibronectin (FN), and then they were transfected with FAK-related non-kinase (FRNK) plasmids mediated by cationic liposome. The apoptosis of FRNK-induced HSC was examined by annexin-V/propidium iodide double-labeled flow cytometry (FCM), gel electrophoresis and transmission electron microscopy. Levels of FRNK, FAK, p-FAK (Tyr397), ERK1 and p-ERK in HSC were assayed by Western blot on the protein level, and by RT-PCR on the mRNA level.</p><p><b>RESULTS</b>The expression of FRNK was enhanced after FRNK plasmids were transiently transfected into the HSC. The apoptotic rate of the HSC exposed to FRNK plasmids for 48 h was higher than that in the non-FRNK plasmid group (25.37%+/-1.92% vs 9.28%+/-1.05%, P less than 0.01), and was accompanied by a significantly higher activity of caspase-3 both in the protein and in the mRNA levels [(264.17+/-12.60 vs 185.82+/-9.69), P less than 0.01; (4.19+/-0.48 vs 1.07+/-0.27), P less than 0.01]. After exposure of HSC to FRNK plasmids, compared with the non-FRNK plasmid group, the expressions of p-FAK, ERK1 and p-ERK in protein and mRNA levels were lower; on the contrary, compared with the control group, the expressions of p-FAK, ERK1 and p-ERK in the FN group were higher.</p><p><b>CONCLUSION</b>The expression of FRNK was enhanced and the phosphorylation of FAK was inhibited after FRNK was transiently transfected into HSC in vitro. FRNK induces apoptosis of HSC. FAK-ERK signal transduction pathway perhaps is involved in the process.</p>

Angiotensin II stimulates platelet-derived growth factor-B expression in hepatic stellate cells by activating EGR-1 / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the signal transduction mechanism underlying the effects of angiotensin II (AngII) on extracellular signal-regulated kinase 1/2 (ERK1/2), early growth response-1 (EGR-1) and platelet-derived growth factor-B (PDGF-B) in hepatic stellate cells (HSCs).</p><p><b>METHODS</b>HSC-T6 cells treated with AngII for 10 or 30 min were examined for phospho-P42/44 protein expression using Western blotting. In another experiment, the cells were preincubated for 1 h in the presence of U0126 (an inhibitor of the MAPK/ERK kinase), irbesartan (an AT-1 receptor blocker), or antioxidant-N-acetylcysteine (NAC) prior to AngII exposure, and the protein expression of phospho-P42/44 and PDGF-B were measured with Western blotting. The DNA binding activity of EGR-1 was analyzed using electrophoretic gel mobility shift assay (EMSA), and the expression of PDGF-B was detected immunohistochemically.</p><p><b>RESULTS</b>AngII induced phospho-P42/44 expression in HSC-T6, which was abrogated by U0126 or irbesartan. NAC did not inhibit phospho-P42/44 expression. EMSA showed that AngII exposure of the HSC cells markedly increased EGR-1 DNA binding activity, reaching the maximum after 60 min of exposure followed by progressive declination; irbesartan and U0126 significantly suppressed AngII-induced EGR-1 activity enhancement. ACEI at 1 micromol/L and 10 nmol/L inhibited EGR-1 activity, but ACEI at the concentration of 0.1 nmol/L resulted in enhanced EGR-1 activity. NAC showed no obvious effect in suppressing EGR-1 activity. AngII increased PDGF-B protein level in the HSCs, the effect of which was inhibited by irbesartan. U0126, NAC and ACEI did not attenuate PDGF-BB protein level in the HSCs.</p><p><b>CONCLUSION</b>Stimulation of the HSCs with AngII results in EGR-1 activation via the ERK1/2 pathway, leading to up-regulation of PDGF-B expression.</p>

Spheres isolated from Colo205 cell line possess cancer stem-like cells under serum-free culture condition / 南方医科大学学报

<p><b>OBJECTIVE</b>Isolation and expansion tumor spheres from colorectal cancer cell line Colo205 cultured in serum-free medium(SFM) supplemented with human recombinant EGF and bFGF.</p><p><b>METHODS</b>Colo205 cells were cultivated in SFM,while cells cultivated in serum-supplemented medium(SSM) served as the control. Cells morphology were observed by optical microscope, and expression of intestinal stem cells marker Musashi-1 was detected by immunocytochemical. To induce cell differentiation, tumour spheres were cultivated without EGF and bFGF in the presence of 10% serum. Then we analysed expressions of stem cell surface markers CD133 and CD44 among undifferentiated cell, post-differentiated cells and routine Colo205 cells under serum-supplemented culture condition by flow cytometry. At last we compared cell cycle and spectral karyotype between two groups.</p><p><b>RESULTS</b>In SFM consisting of EGF and bFGF, a minority of Colo205 cells could survive, proliferate and form the suspended tumor spheres. We detected high Musashi-1 expression in these cells. Compared with the SSM group and the post-differentiation SFM group, the expressions of CD133 and CD44 were significantly increased in the undifferentiated SFM group (P<0.05). There was no statistical difference in the expression of CD133 and CD44 between the post-differentiation SFM group and the SSM group (P>0.05). Cell cycle analysis indicated that tumor spheres were of a high proliferation state.We could not find any noticeable difference in the number of chromatosomes between the SFM group and the SSM group.</p><p><b>CONCLUSION</b>Tumor spheres in which enriched cancer stem cells can be generated under serum-free culture condition with EGF and bFGF.</p>

The effects of FRNK on expressions of MMP-2 mRNA and TIMP-2 mRNA in hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the effects of FAK-related non-kinase (FRNK) on expressions of type I collagen and matrix metalloproteinase-2 (MMP-2) mRNA and tissue inhibitor of metalloproteinase-2 (TIMP-2) mRNA in rat hepatic stellate cells (HSC).</p><p><b>METHODS</b>Using in vitro cell culture technique, FRNK plasmids were transfected into HSC mediated by cationic liposome. Type I collagen synthesis capability in HSC was examined by 3H-Pro incorporation assay. The levels of FRNK in HSC were assayed by Western blot, and the expressions of MMP-2 and TIMP-2 were assayed by RT-PCR on mRNA levels.</p><p><b>RESULTS</b>The exposure of HSC to FRNK caused the expression of FRNK protein to be up-regulated, and the FRNK protein contents reached the highest point at 48 h after the transfection, P less than 0.05. The expressions of MMP-2 mRNA were up-regulated by FRNK; the expressions of TIMP-2 mRNA were down-regulated by FRNK; the ratios of MMP-2 mRNA/TIMP-2 mRNA were enhanced by FRNK.</p><p><b>CONCLUSION</b>After FRNK was transfected, the capability of type I collagen synthesis in HSC was inhibited, which may be related to the up-regulation of MMP-2 mRNA/TIMP-2 mRNA.</p>

The dynamic expression of PTEN in fibrogenic rat liver tissues and its relation to the activation and proliferation of hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the dynamic expression of PTEN in fibrogenic liver tissue of rats and its effect on the activation and proliferation of hepatic stellate cells (HSC).</p><p><b>METHODS</b>A rat model of hepatic fibrosis was established by common bile duct ligation (BDL). The expressions of PTEN in the rat liver tissues were detected by immunohistochemical staining, Western blot and real-time PCR assay. The expressions of PTEN in activated HSC in the rat liver tissues were detected by immunofluorescence double labeling confocal laser scanning microscopy. The alpha-SMA in the rat liver tissues was determined by immunohistochemical staining.</p><p><b>RESULTS</b>The immunohistochemical staining indicated that there was extensive expression of PTEN in the liver tissues of normal rats, it was expressed mainly in the cytoplasm of the HSC. With the aggravation of hepatic fibrosis, the expression of PTEN in the hepatic tissues decreased gradually (P less than 0.01), while the alpha-SMA positive cells in the hepatic tissues increased significantly (P less than 0.01). The expressions of PTEN protein and mRNA in the rat liver tissues at week 1, 2, 3 and 4 after BDL were all lower than those in the sham operation group (P less than 0.01), and the expressions gradually decreased with the development of hepatic fibrosis (P less than 0.01). Immunofluorescence double labeling confocal laser scanning microscopy showed that PTEN were expressed extensively in activated HSC, especially in the cytoplasm, and with the development of hepatic fibrosis, the PTEN-expressing activated HSC accounted for an increasingly smaller percentage of total activated HSC.</p><p><b>CONCLUSION</b>The expressions of PTEN mRNA and protein in rat fibrogenic liver tissues were downregulated, and their expressions in HSC in vivo also decreased. The dynamic expressions of PTEN in liver tissues had a significant negative correlation with the activation and proliferation of HSC.</p>

Effects of Changyanqing decoction on the expressions of interleukin-10 and intercellular adhesion molecule-1 in rats with ulcerative colitis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of Changyanqing decoction, a traditional Chinese medicinal preparation, on the expressions of interleukin-10 (IL-10) and intercellular adhesion molecule-1 (ICAM-1) in the colon mucosa of rats with ulcerative colitis.</p><p><b>METHODS</b>The rats with ulcerative colitis induced by trinitrobenzene sulphonic acid and ethanol enema were randomly divided into 3 groups, namely the model group, sulfasalazine (SASP) group, and Changyanqing decoction group. Daily treatment with intragastric administration and enema of normal saline, SASP (100 mg/kg), and Changyanqing decoction (39.75 mg/kg), respectively, were administered 24 h after the establishment of colitis till the end of the experiment. Another group of rats was used as the normal control group. The disease activity index (DAI) and colon mucosa damage index (CMDI) of the rats were calculated. The activity of myeloperoxidase (MPO) was measured by biochemical method, and the expressions of IL-10 and ICAM-1 protein were measured by ELISA and immunohistochemistry, respectively.</p><p><b>RESULTS</b>Compared with the normal group, the model group showed significantly increased DAI, CMDI, HS score and MPO activity in the colon tissues (P < 0.01), with also significantly increased expression of ICAM-1 (P < 0.01) and decreased expression of IL-10 in the rat colon mucosa (P < 0.01). Treatment with Changyanqing decoction resulted in a significant reduction in DAI, CMDI, HS score and MPO activity (P < 0.01), and decreased the expression of ICAM-1 (P < 0.01) and increased the expression of IL-10 (P < 0.01) in the colon mucosa. The expression of ICAM-1 in the colon mucosa was positively correlated to that of IL-10 (r = 0.927, P < 0.01) and the activity of MPO (r = 0.621, P < 0.01).</p><p><b>CONCLUSIONS</b>Changyanqing decoction has protective effect against rat ulcerative colitis, mediated probably by enhancement of IL-10 expression and reduction in ICAM-1 expression and neutrophil infiltration.</p>

Effect of angiotensin II on Rho-Rock pathway in rat hepatic stellate cell contraction / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the mechanisms of angiotonin II (AngII)-induced Ca(2+)-independent pathways mediated by Rho kinase in hepatic stellate cells (HSCs).</p><p><b>METHODS</b>HSC-T6 cells were treated with 1 micromol/L of AngII, and the subsequent cell contraction was directly observed with silicone rubber membrane culture method. The cells with 10 micromol/L AngII treatment were examined for myosin light chain (MLC) phosphorylation level using Western blotting, and the effects of irbesartan (a specific inhibitor of AngII 1- receptor) and Y27632 (a Rho kinase inhibitor) on AngII-induced MLC phosphorylation were evaluated. RT-PCR was used to detect the expression of Rock2 in Ca(2+)- independent pathways mediated by Rho kinase.</p><p><b>RESULTS</b>AngII induced HSC contraction and time-dependent MLC phosphorylation changes, which peaked 15 min after the treatment followed by gradual reduction. Irbesartan or Y27632 treatment significantly lowered MLC phosphorylation level in AngII-induced cells (P<0.01). The mRNA expression of Rock2 increased significantly after AngII treatment (P<0.01), but decreased following subsequent irbesartan or Y27632 treatment.</p><p><b>CONCLUSION</b>AngII induces HSC contraction through Ca(2+)-independent pathways mediated by Rho kinase.</p>