Effect of Hic-5 gene knockout on NF-κB/p65 expression and CCl4-induced liver fibrosis degree in mice / 临床肝胆病杂志

@#ObjectiveTo investigate the effect of Hic-5 gene knockout on NF-κB/p65 expression and liver fibrosis. MethodsTen wild-type male C57BL/6 mice were randomly divided into wild-type control group (WT-Control group with 5 mice) and wild-type experimental group (WT-CCl4 group with 5 mice), and ten male C57BL/6 mice with Hic-5 gene knockout were randomly divided into Hic-5 knockout control group (Hic-5 KO-Control group with 5 mice) and Hic-5 knockout experimental group (Hic-5 KO-CCl4 group with 5 mice). The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured. Picrosirius red staining was used to observe collagen deposition in liver tissue. Immunohistochemistry was used to measure the expression of alpha-smooth muscle actin (α-SMA) and p65 protein, and real-time quantitative PCR was used to measure the mRNA expression of α-SMA, collagen 1, and p65 in liver tissue. The primary hepatic stellate cells of mice were isolated and stimulated with different concentrations of TGF-β1, and then real-time quantitative PCR was used to measure the mRNA expression of α-SMA, collagen 1, and p65 in primary hepatic stellate cells. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsPicrosirius red staining showed that compared with the WT-CCl4 group, the Hic-5 KO-CCl4 group had a significant reduction in collagen fibers in liver tissue (P＜0.001). Measurement of serum ALT and AST showed that there were significant differences in ALT and AST between the WT-Control group, the WT-CCl4 group, the Hic-5 KO-Control group, and the Hic-5 KO-CCl4 group (F=22.85 and 25.15, both P＜0.001), and the Hic-5 KO-CCl4 group had significantly lower serum levels of ALT and AST than the WT-CCl4 group (both P＜0.05). Immunohistochemistry showed that there were significant differences in the expression levels of α-SMA and p65 protein in liver tissue between the WT-Control group, the WT-CCl4 group, the Hic-5 KO-Control group, and the Hic-5 KO-CCl4 group (F=207.10 and 98.16, both P＜0.001), and the Hic-5 KO-CCl4 group had significantly lower expression of α-SMA and p65 protein in liver tissue than the WT-CCl4 group (both P＜0.01). The results of real-time quantitative PCR showed that there were significant differences in the relative mRNA expression of α-SMA, collagen 1, and p65 in liver tissue between the WT-Control group, the WT-CCl4 group, the Hic-5 KO-Control group, and the Hic-5 KO-CCl4 group (F=41.62, 13.93, and 98.16, all P＜0.001), and the Hic-5 KO-CCl4 group had significantly lower relative mRNA expression of α-SMA, collagen 1, and p65 in liver tissue than the WT-CCl4 group (all P＜0.05). After the primary hepatic stellate cells were stimulated by TGF-β1 at concentrations of 0, 5, and 10 ng/ml, there were significant differences in the relative mRNA expression of α-SMA, collagen 1, and p65 between the WT 0 ng/ml group, the WT 5 ng/ml group, the WT 10 ng/ml group, the KO 0 ng/ml group, the KO 5 ng/ml group, and the KO 10 ng/ml group (F=53.9, 75.82, and 52.41, all P＜0.001), and the Hic-5 KO group had significantly lower relative mRNA expression of α-SMA, collagen 1, and p65 than the WT group (all P＜0.01). ConclusionHic-5 knockout inhibits NF-κB/p65 expression and hepatic stellate cell activation and alleviates CCl4-induced liver fibrosis.

Synergistic effect of RhBMP-2 and bFGF on ectopic osteogenesis in mice

OBJECTIVE@#To investigate the synergistic effect and mechanism of the combined application of recombinant human bone morphogenetic protein-2 (rhBMP-2) and basic fibroblast growth factor (bFGF).@*METHODS@#24 KM male mice were randomly divided into 6 groups with 4 mice in each group, namely, Group A (control group), Group B (only treated with collagen), Group C (treated with 2 ng bFGF+collagen), Group D (treated with 4 μ g rhBMP-2+collagen), Group E (treated with 4 μ g rhBMP-2+2 ng bFGF+collagen) and Group F (treated with 4 μ g rhBMP-2+4 ng bFGF+collagen). The composites were implanted into the intermuscular septum of hind legs mice; whereas in control group, intermuscular septum of mice was separated and no implantation was performed. General observation, detection of concentration of calcium content, micro computed tomography (Micro-CT), three-dimensional reconstruction scan, measurement of bone mineral density (BMD), bone volume fraction (BVF) and trabecular thickness (Tb.Th), as well as histological observation with HE staining and ALP and CD34 immumohistochemical staining were performed.@*RESULTS@#Ectopic osteogenesis was found in Groups D, E and F mice. The difference in concentration of calcium contents was statistically significant between Groups D and E (P0.05). Micro-CT and three-dimensional reconstruction revealed continuous newborn bone substance in external surface of ectopic bone formation, and the center of bone formation did not show obvious substantial filling by bone substance. The differences in BMD, BVF and Tb.Th were statistically significant between Groups D and E or F (P<0.01 or <0.05). HE staining showed that in Groups D, E and F, newborn bone substance was mainly located at the edge of ectopic bone formation, and the bone formation in Groups E and F was better than that in Group D. ALP and CD34 immumohistochemical staining revealed the positive expression mainly at the edge of ectopic bone formation, and area of positive expression in Groups E and F was larger than that in Groups D.@*CONCLUSIONS@#rhBMP-2 possesses the capacity to induce ectopic osteogenesis independently, but bFGF does not have this ability; the combined application of rhBMP-2 and bFGF can enhance the synergetic effect on inducing ectopic osteogenesis.

Synergistic effect of RhBMP-2 and bFGF on ectopic osteogenesis in mice

Objective: To investigate the synergistic effect and mechanism of the combined application of recombinant human bone morphogenetic protein-2 (rhBMP-2) and basic fibroblast growth factor (bFGF). Methods: 24 KM male mice were randomly divided into 6 groups with 4 mice in each group, namely, Group A (control group), Group B (only treated with collagen), Group C (treated with 2 ng bFGF+collagen), Group D (treated with 4 μ g rhBMP-2+collagen), Group E (treated with 4 μ g rhBMP-2+2 ng bFGF+collagen) and Group F (treated with 4 μ g rhBMP-2+4 ng bFGF+collagen). The composites were implanted into the intermuscular septum of hind legs mice; whereas in control group, intermuscular septum of mice was separated and no implantation was performed. General observation, detection of concentration of calcium content, micro computed tomography (Micro-CT), three-dimensional reconstruction scan, measurement of bone mineral density (BMD), bone volume fraction (BVF) and trabecular thickness (Tb.Th), as well as histological observation with HE staining and ALP and CD34 immumohistochemical staining were performed. Results: Ectopic osteogenesis was found in Groups D, E and F mice. The difference in concentration of calcium contents was statistically significant between Groups D and E (. P0.05). Micro-CT and three-dimensional reconstruction revealed continuous newborn bone substance in external surface of ectopic bone formation, and the center of bone formation did not show obvious substantial filling by bone substance. The differences in BMD, BVF and Tb.Th were statistically significant between Groups D and E or F (. P<0.01 or <0.05). HE staining showed that in Groups D, E and F, newborn bone substance was mainly located at the edge of ectopic bone formation, and the bone formation in Groups E and F was better than that in Group D. ALP and CD34 immumohistochemical staining revealed the positive expression mainly at the edge of ectopic bone formation, and area of positive expression in Groups E and F was larger than that in Groups D. Conclusions: rhBMP-2 possesses the capacity to induce ectopic osteogenesis independently, but bFGF does not have this ability; the combined application of rhBMP-2 and bFGF can enhance the synergetic effect on inducing ectopic osteogenesis.

Low-density lipoprotein cholesterol target goal attainment rate and related factors in patients with acute coronary syndrome after percutaneous coronary intervention / 中华心血管病杂志

<p><b>OBJECTIVE</b>To observe the low-density lipoprotein cholesterol (LDL-C) target goal attainment rate and related factors in patients with acute coronary syndrome (ACS) after percutaneous coronary intervention (PCI).</p><p><b>METHODS</b>From March 2011 to March 2012, a total of 832 ACS patients were retrospectively evaluated in the Cardiology Department of the First Affiliated Hospital of Dalian Medical University. The target goal attainment rate after PCI was defined as the percentage of patients reaching LDL-C goals recommended by The European Society of Cardiology (ESC) and the European Atherosclerosis Society (EAS) guidelines for the management of dyslipidemias (European guidelines) and Chinese guidelines on prevention and treatment of dyslipidemias in adults and Chinese guidelines on percutaneous coronary artery intervention treatment (Chinese guidelines). Multivariate logistic regression analysis was used to analyze the related factors.</p><p><b>RESULTS</b>According to the European guidelines, the overall LDL-C goal attainment rates at 1 month and 9 months after PCI were 25.2% (210/832) and 22.2% (186/832), respectively. According to the Chinese guidelines, the overall LDL-C goal attainment rates at 1 month and 9 months after PCI were 46.5% (387/832) and 42.3% (352/832), respectively. In accordance with the Chinese guidelines, the multivariate logistic regression analysis showed that gender (females/males, OR = 0.650, 95%CI: 0.442-0.956), age ( ≥ 60 years/<60 years, OR = 0.628, 95%CI:0.464-0.850), hypertension (OR = 0.737, 95%CI: 0.547-0.994), prior myocardial infarction history (OR = 0.696, 95%CI:0.511-0.948), prior PCI history (OR = 0.575, 95%CI: 0.339-0.974) and baseline LDL-C levels ( OR = 0.155, 95%CI: 0.096-0.252) were independent risk factors that affected LDL-C goal attainment at 1 month post PCI. Moreover, the following parameters were the independent risk factors for LDL-C goal attainment at 9 months after PCI: prior myocardial infarction history (OR = 0.706, 95%CI:0.521-0.958), prior PCI history (OR = 0.565, 95%CI:0.334-0.957) and baseline LDL-C levels (OR = 0.176, 95%CI:0.110-0.282).</p><p><b>CONCLUSIONS</b>Currently, the LDL-C control rate is low in patients with ACS after PCI. The cholesterol lowering therapy should be individually strengthened for patients after PCI, especially in female patients, patients with aged ≥ 60 years old, hypertension, prior myocardial infarction history, prior PCI history and higher baseline LDL-C level.</p>

Effects of plasma Hcy on the contents of CO and HO-2 in the corpus cavernosum of ED rats with hyperhomocysteinemia / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the changes in the activities of carbon monoxide (CO) and heme oxygenase 2 (HO-2) in ED rats with hyperhomocysteinemia (HHcy).</p><p><b>METHODS</b>This study included 40 male Wistar rats weighing 280 - 310 g, 10 as normal controls (group A). HHcy models were made in the other 30 by giving 3% methionine for 4 weeks, and then divided into groups B, C and D. The rats in group B continued to be fed with 3% methionine, those in group C were treated with betaine hydrochloride, and those in group D were given zinc porphyrin IX at 45 micromol per kg per d. Penile erections of the rats were recorded, and 4 weeks later, all were killed for determination of the levels of homocysteine (Hcy) in the blood plasma and the activities of CO and HO-2 in the corpus cavernosum of the penis.</p><p><b>RESULTS</b>The level of plasma Hcy, penile erection frequency and the content of CO in the corpus cavernosum were (12.55 +/- 0.82) micromol/L, (1.88 +/- 0.05) times and (10.55 +/- 1.73) micromol/L in group A, the Hcy level significantly higher while the penile erection frequency and CO content remarkably lower than in group B ([25.01 +/- 0.94] micromol/L, [0.70 +/- 0.05] times and [9.51 +/- 1.52] micromol/L, P < 0.05 or P < 0.01), with a negative correlation between the level of Hcy and that of CO and HO-2 (P < 0.01). Compared with group B, the three parameters were all significantly increased in C ([14.37 +/- 0.47] micromol/L, [1.18 +/- 0.08] times and [10.36 +/- 1.56] micromol/L, all P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>Decreased expressions of CO and HO-2 in the corpus cavernosum of the penis may result in ED in HHcy rats. Betaine can reduce the Hcy level in the blood plasma and CO content in the corpus cavernosum, which might be one of the mechanisms of its action on ED with HHcy.</p>

Application of ultrasonic inspection in monitoring dynamic healing of mandibular fracture in rabbit model

OBJECTIVE@#To investigate the feasibility of ultrasonic diagnosis for monitoring fracture healing.@*METHODS@#Thirty rabbit models with fraction of mandible body were established by surgically removing partial lower jawbone. At the 1st, 2nd, 4th, 6th, 8th and 12th week after the operation, they were examined by X-ray and ultrasound, respectively. All detection results were scored according to a generally accepted standard. Spearman rank correlation analysis was conducted to explore the relationship between the results of the two inspection methods.@*RESULTS@#In each healing stage, the results of the ultrasonic inspection were basically consistent with those of the X-ray examination, as supported by a Spearman rank correlation coefficient of 0.892 (P<0.001).@*CONCLUSIONS@#Non-invasive ultrasonic inspection can be used instead of X-ray examination to monitor and diagnose fracture healing.

Inflammation inhibitory effects of sirolimus and paclitaxel-eluting stents on interleukin-1β-induced coronary artery in-stent restenosis in pigs / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Coronary artery in-stent restenosis (ISR) and late stent thrombosis remain as important complications of stenting. The inflammation reactions to sirolimus and paclitaxel-eluting stents were investigated in a swine stenosis model induced by interleukin (IL)-1β.</p><p><b>METHODS</b>Mini pigs (n = 12; 2-3 months old and weighing 25-30 kg) were subjected to thoracotomy. Segments (10 mm) of the mid left anterior descending coronary artery and left circumflex coronary artery were exposed and aseptically wrapped with a cotton mesh soaked with IL-1β (5 µg). After 2 weeks, the animals were anesthetized and quantitative coronary arteriography (QCA) was performed. The stenosis sites were randomized into three groups for stent insertion: a sirolimus-eluting stent (SES) group (Firebird(TM), n = 7), a paclitaxel-eluting stent (PES) group (TAXUS(TM), n = 9), and a bare-metal stent (BMS) group (YINYITM, Dalian Yinyi Biomaterials Development Co., Ltd, China, n = 8). The three different stents were randomly implanted into stenosis segments. Expression of monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-alpha (TNF-α), P-selectin and vascular cell adhesion molecule-1 (VCAM-1) was determined by reverse transcription-coupled polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>QCA showed severe stenosis in IL-1β treated segments. The SES and PES groups showed lower 1-month angiographic late lumen loss (LLL) within the stent and the lesion compared with BMS (P < 0.05) by follow-up QCA. The SES showed lower LLL than that of PES in reducing 1-month inflammation lesions in pigs by follow-up QCA ((0.15 ± 0.06) mm vs. (0.33 ± 0.01) mm, P < 0.0001). The neointimal hyperplasia areas in SES and PES showed lower than those of BMS (SES (11.6 ± 1.7) mm(2), PES (27.2 ± 1.6) mm(2) vs. BMS (76.2 ± 1.3) mm(2), P < 0.0001). The mRNA expression of MCP-1 by RT-PCR in SES and PES showed lower than that of BMS at 30 days after stenting (SES 0.20 ± 0.03, PES 0.48 ± 0.49 vs. BMS 0.58 ± 0.07, P < 0.05). Levels of VCAM-1 in SES were significantly lower than those of PES and BMS (SES 0.35 ± 0.08 vs. PES 0.65 ± 0.13, BMS 0.70 ± 0.06, P < 0.05). Histochemical immunostaining of vessel walls showed lower inflammatory chemokine MCP-1 expression in the SES and PES groups compared with BMS.</p><p><b>CONCLUSION</b>SESs were superior in reducing 1-month angiographic LLL in inflammation lesions in pigs, strongly suggesting that SESs can suppress inflammatory reactions in ISR at multiple points.</p>

Expression of B lymphocyte stimulator in peripheral blood mononuclear cells in individuals with systemic lupus erythematosus and the role of interferon-? on it's expression / 中华风湿病学杂志

Objective To determine the expression of membrane-bound B lymphocyte stimulator (BLyS) protein and its mRNA in vitro of peripheral blood mononuclear cells (PBMCs) from individuals with systemic lupus erythematosus (SLE),and to investigate the role of interferon-?(IFN-?) on the expression of BLyS.Methods PBMCs were obtained from 25 SLE patients (mean age of 31+14) and 20 healthy volunteers (mean age of 28?10).They were randomized into IFN-?(5 ng/ml) group and control group.PBMCs were col- lected at 0,6,12 and 24 h for BLyS mRNA assessment using semi-quantitative reverse transcription-PCR (RT-PCR).PBMCs were also collected at 72 h for membrane-bound BLyS protein detection using flow cy- tometry (FACS) and direct immunofluorescence.Results①The expression of BLyS mRNA and membrane- bound protein in PBMCs was significantly higher in individuals with SLE compared with healthy controls (P＜0.05);②IFN-?enhanced BLyS mRNA expression in PBMCs in both healthy controls and SLE patients,with the greatest effect at 6 h (stimulated vs unstimulated,0.42?0.19 vs 0.25?0.14,P＜0.01;0.59?0.28 vs 0.44?0.21,P＜0.01 );③IFN-?also increased the expression of membrane-bound BLyS protein in both healthy con- trols and individuals with SLE (FACs,mean fluorescence intensity,4.5+3.0 vs 3.7~2.6,P

Acquired hemophilia A in patients with systemic lupus erythematosus:report of a case and review of literature / 中华风湿病学杂志

Objective To summarize the manifestation and treatment of acquired hemophilia A in pa- tients with systemic lupus erythematosus(SLE).Method A case was investigated retrospectively and the lit- erature was reviewed.Results A 25-year-old woman with a 5 year history of SLE was admitted to hospital due to abdominal pain.She was diagnosed with acquired factorⅧinhibitor deficiency based on a prolonged activated partial-thromboplastin time(APTT,135.3 s),reduced factorⅧactivity(0.9%)and factorⅧin- hibitor(26.1 BU/ml).Sonography and magnetic nuclear resonance of the abdomen confirmed the presence of a retro-uterine hematoma.The patient was initially treated with intravenous pulse and oral corticosteroids,factorⅧplasma concentrated and intravenous immunoglobulin.Clinical and biological improvement was promptly obtained.Conclusions Attention should be paid to the association between SLE and acquired hemophilia A. Combination therapy may be recommended as initial therapy for the management of acquired hemophilia A in patients with SLE.But no standardized treatment can be recommended at present.