Histone acetyltransferases (HATs) are involved in the regulation of gene transcription in eukaryotic cells and their inhibitors could be a promising class of drugs due to their ability to modulate transcription and exert antiviral, anti-inflammatory as well as antioxidant effects. Nonradioactive spectrophotometric HAT assay is an alternative method to the widespread radioactive assay but suffers from drawbacks as lack of sensitivity and accuracy. A simple, non-radioactive fluorescent assay that measures the production of CoASH was established by its facile reaction with O-phthalaldehyde and 2-amino-ethanol. This method gives much higher accuracy compared to spectrophotometric assay, and allows screening of various compounds with potential HAT inhibition. The novel assay should be a valuable tool in transcriptional research and especially drug discovery.

BACKGROUND: Connective tissue growth factor (CTGF) has very strong regulative effect on the differentiation and proliferation of renal cells as well as the synthesis and degradation of extracellular matrix. Its abnormal expression plays a key role in the process of fibrous degeneration of various renal diseases including diabetic nephropathy.OBJECTIVE: To observe the protective effect ofjiedu tongluo baoshen gelatin capsule on renal disease in rats with diabetes mellitus and its effect on the expression of renal CTGF mRNA.DESIGN: A randomized controlled experiment taking animals as subjects.SETTING: Institute of Traditional Chinese Medicine, Changchun College of Traditional Chinese Medicine; Department of Pathology, Institute of Regenerated Medical Sciences, Jilin University MATERIALS: Totally 46 Wistar rats, of clean degree, male and female in half, weighing 150 to 200 g, were used in the experiment. Jiedu tongluo baoshen gelatin capsule was provided by Preparation Room, Traditional Chinese Medicine Hospital of Jilin Province (Batch No. 2002101, composed of Renshen, Huangqi, Shengdi, Gouqizi, Zhimuv Digupiv Huanglian,Zhenhua, Jinyinhua, Dahuang, Danshen, Yimucao and so on, 0.25 g/pill)and benazepril (Provided by Beijing Nuohua Pharmaceutical Company,Batch No. 01046) were used in the experiment.METHODS: This experiment was carried out in the Department of Pathology, Institute of Regenerated Medical Sciences, Jilin University from May 2002 to January 2003. ① Totally 46 Wistar rats used in the experiment were randomly divided into 4 groups: normal control group (n=10), model group (n=12), benazepril-treated group (n=12) and traditional Chinese medicine-treated group (n=12).Rats of normal control group were intraperitoneally injected of natrium citricum buffer solution of same volume; Rats of other groups were intraperitoneally injected of 50 mg/kg streptozotocin which was dissolved by 0.1 mol/L citric acid buffer solution before use(pH 4.2). Seventy-two hours after modeling, venous blood was collected from the tails to measure the levels of fasting blood glucose and urine glucose.Blood glucose ≥ 16.7 mmol/L and urine glucose over 卅 suggested diabetic models were successful. Rats of benazepril-treated group were performed intragastric administration of benazepril suspension of 15 mg/(kg·d);Rats of traditional Chinese medicine-treated grou p were performed intragastric administration ofjiedu tongluo baoshen gelatin capsule suspension of 5 g/(kg ·d); Rats of normal control group and model group were performed intragastric administration of the same volume of running water on each morning, once a day, 12 weeks in total. ② Levels of blood glucose,urea nitrogen, serum creatinine and urinary albumin excretion rate were measured 12 weeks after administration; Level of angiotensin Ⅱ of renal tissue was measured with radioimmunoassay; Levels of renal fibronectin and type Ⅳ collagen protein expression were measured with immunohistochemical method; Connective tissue growth factor gene expression of renal cortex was measured with reverse transcription-polymerase chain reaction and renal histological observation was also performed. ③ Analysis of variance was used in the intergroup comparison.MAIN OUTCOME MEASURES: Biochemical index of blood and urine,level of angiotensin Ⅱ of renal tissue, pathological morphology, expression of fibronectin and type Ⅳ rabbit anti-rat collagen protein and expression of connective tissue growth factor mRNARESULTS: Five, four and three rats died in the process of experiment respectively in model group, benazepril-treated group and traditional Chinese medicine-treated group, finally, 10,7,8 and 9 rats entered the stage of result analysis respectively in normal control group, model group, benazepril-treated group and traditional Chinese medicine-treated group. ① The ratio between body mass and renal mass was significantly higher in the model group than in the normal control group (P ＜ 0.05),but was significantly lower in the benazepril-treated group and traditional Chinese medicine-treated group than in the model group (P ＜ 0.01). ② Fasting blood glucose was significantly higher in the model group than in the normal control group (P ＜ 0.01),but was significantly lower in the traditional Chinese medicine-treated group than in the model group and benazepril-treated group (P ＜ 0.01). ③ Levels of urea nitrogen, serum creatinine, creatinine clearance, urinary albumin excretion rate and angiotensin Ⅱ of renal tissue were significantly higher in the model group than in the normal control group (P ＜ 0.01), but significantly lower in the benazepril-treated group and traditional Chinese medicine-treated group than in the model group (P ＜ 0.05-0.01 ). ④Pathomorphological change of renal tissue was very severe in the model group,but very mild in the benazepril-treated group and traditional Chinese medicine-treated group. ⑤Fibronectin and type Ⅳ rabbit anti-rat collagen protein expression were significantly higher in the model group than in the normal control group (P ＜ 0.01), those were significantly lower in the benazepril-treated group and traditional Chinese medicine-treated group than in the model group, but higher than normal control group (P ＜ 0.01). ⑥Expression of CTGF mRNA of renal cortex of model group was 2.65 times that of normal control group. Expression of CTGF mRNA of benazepriltreated group and traditional Chinese ine-treated group was decreased by 23.68% and 26.32%, respectively (P ＜ 0.05 or 0.01).CONCLUSION: Jiedu tongluo baoshen gelatin capsule has protective effect on diabetic nephropathy, which might be implemented by reducing the level of angiotensin Ⅱ of renal tissue and down-regulating the expression of CTGF mRNA of renal cortex of rats with diabetes mellitus, so as to reduce the deposition of extracellular matrix.

Objective To observe the clinical efficacy of thunder-fire moxibustion at Dazhui (GV14) and Shenshu (BL23) in treating mild cognitive impairment (MCI) due to ischemic cerebral stroke. Method Sixty patients with MCI due to ischemic cerebral stroke were randomized into a treatment group and a control group, 30 cases each. The control group was intervened by joint treatment plus cognitive training, while the treatment group was by thunder-fire moxibustion in addition to that given to the control group. The two groups were evaluated by the Montreal Cognitive Assessment (MoCA), Mini-Mental State Examination (MMSE) and Wechsler Memory Scale (WMS), the major symptoms were observed, and the clinical efficacies were compared between the two groups. The two groups were both treated for 8 weeks. Result The total effective rate was 80.0%in the treatment group, versus 70.0% in the control group, and the difference was statistically significant(P<0.05). Conclusion The selected acupoints can improve the cognition and activities of daily living (ADL) in MCI patients;thunder-fire moxibustion at Dazhui and Shenshu plus cognitive training can produce a better clinical efficacy than dry cognitive training. Therefore, we can combine thunder-fire moxibustion with modern rehabilitation to enhance the therapeutic efficacy in preventing and treating MCI.

Objective To investigate the clinical efficacy of Nazi method of midnight-midday ebb flow plus warm needling moxibustion in treating transient ischemic attack (TIA). Methods Sixty patients with transient ischemic attack were randomly allocated to treatment and control groups, 30 cases each. The control group received routine medication and the treatment group, Nazi method of midnight-midday ebb flow plus warm needling moxibustion in addition. The main clinical symptoms were observed by TCD monitoring of microembolic signals in the two groups before and after treatment. The clinical therapeutic effects were compared between the two groups. Results After treatment, the positive rate of microembolus monitoring was 3.3%in the treatment group and 20.0%in the control group;there was a statistically significant difference between the two groups (P<0.05). There was a statistically significant pre-/post-treatment difference in the traditional Chinese medical syndrome score in the two groups (P<0.05). There was a statistically significant post-treatment difference in the traditional Chinese medical syndrome score between the treatment and control groups (P<0.05). The total efficacy rate was 96.7% in the treatment group and 80.0% in the control group;there was a statistically significant difference between the two groups (P<0.05). At six months after treatment, the rate of disappearance of TIA, the rate of recurrence of TIA and the rate of occurrence of cerebral infarction were 90.0%, 6.7%and 3.3%, respectively, in the treatment group and 60.0%, 20.0%and 20.0%, respectively, in the control group;there were statistically significant differences between the two groups (P<0.05). Conclusions Nazi method of midnight-midday ebb flow plus warm needling moxibustion can decrease the positive rate of microemboli and shorten transient ischemic attack to prevent cerebral infarction in patients with transient ischemic attack.

The aim of this study is to prepare hyaluronic acid (HA) modified core-shell liponanoparticles (pHA-LCS-NPs) as gene delivery system and investigate its gene transfection efficiency in human retinal pigment epithelium (ARPE-19) cells in vitro. The pHA-LCS-NPs was prepared by firstly hydrating dry lipid film with CS-NPs suspension to get LCS-NPs, then modifying the lipid bilayer with HA by amidation reaction between HA and dioleoyl phosphatidylethanolamine (DOPE). Its morphology, particle size and zeta potential were investigated. XTT assay was used to evaluate the cell safety of different vectors in vitro. The gene transfection efficiency of pHA-LCS-NPs modified with different contents of HA was investigated in ARPE-19 cells with green fluorescent protein (pEGFP) as the reporter gene. The results showed that the obtained pHA-LCS-NPs exhibited a clear core-shell structure with the average particles size of (214.9 +/- 7.2) nm and zeta potential of (-35 +/- 3.7) mV. The 24 h cumulative release of gene from pHA-LCS-NPs was less than 30%. After 48 h incubation, gene transfection efficiency of pHA-LCS-NPs/pEGFP was 1.81 times and 3.75 times higher than that of CS-NPs/pEGFP and naked pEGFP, respectively. Also no obvious cytotoxicity was observed on pHA-LCS-NPs. It suggested that the pHA-LCS-NPs might be promising non-viral gene delivery systems with high efficiency and low cytotoxicity.
Cell Survival ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Hyaluronic Acid ; chemistry ; pharmacology ; Lipids ; Nanoparticles ; Particle Size ; Phosphatidylethanolamines ; chemistry ; pharmacology ; Retinal Pigment Epithelium ; drug effects ; Transfection

Crocetin, a naturally occurring carotenoid, possesses antioxidant and antiatherosclerotic properties, of which the underlying mechanism remains unclear. In the present study, we examined the effects of crocetin (0.1, 1, 10 μmol·L(-1)) on angiotensin II (Ang II, 0.1 μmol·L(-1)) induced expression of vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vein endothelial cells (HUVECs) and monocyte-endothelial cell adhesion. The effects of crocetin on the activation of nuclear factor kappa B (NF-κB) and intracellular reactive oxygen species (ROS) were also observed. The results demonstrated that crocetin notably suppressed Ang II induced NF-κB activation (P<0.01) and VCAM-1 expression (P<0.05, P<0.01) in HUVECs, accompanied by a markedly reduced monocyte-endothelial cell adhesion (P<0.05, P<0.01). In addition, preincubation with crocetin resulted in a significant enhancement of cellular antioxidant capacity (P<0.05, P<0.01), while Ang II induced intracellular ROS decreased markedly (P<0.05, P<0.01). These results indicated that crocetin was capable of suppressing Ang II induced VCAM-1 expression and monocyte-endothelial cell adhesion by suppression of NF-κB activation, which might be derived from the enhancement of antioxidant capacity and subsequent reduction of intracellular ROS.
Angiotensin II ; metabolism ; Antioxidants ; pharmacology ; Carotenoids ; pharmacology ; Cell Adhesion ; drug effects ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Monocytes ; cytology ; NF-kappa B ; metabolism ; Reactive Oxygen Species ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism

Background With the development of refractive surgery,phakic intraocular lens implantation (PIOL) for high myopia is proving its outstanding merits in short-term treating outcomes.However,its long-term safety and effectiveness were still in more attention.Objective This study was to evaluate and compare the long-term safety,stability and efficacy of Verisyse PIOL implantation with implantable contact lens (ICL) implantation for high myopia.Methods The clinical data of 18 eyes (9 patients) who received Verisyse iris-claw intraocular lens implantation for high myopia and matched 22 eyes (11 patients) who received ICL implantation for high myopia from 2009 to 2011 were retrospectively analyzed.The patients were followed-up for 2 years after surgery.Uncorrected visual acuity (UCVA),best corrected visual acuity (BCVA),spherical equivalence (SE),axial length,corneal endothelial cell density (ECD),contrast sensitivity (CS),wave-front aberrations and postoperative complications were recorded during the following-up.The efficacy index,predictable and safety index were compared between the two groups.Results There was no significant difference in UCVA,BCVA and SE between Verisyse group and ICL group(t =0.92,1.32,4.32;all at P＞0.05).Non-glare CS under the 1.5,3.0,6.0,12.0 and 18.0 c/d spatial frequencies were insignificantly different between the two groups(Z=0.782,0.956,0.495,0.874,0.293;all at P＞0.05),and the similar outcomes were found in glare CS (Z=0.985,1.254,0.896,1.652,0.492;all at P＞0.05) in postoperative 2 years.Root meas square of total high order aberration (RMSh),vertical-trefoil,vertical-coma,horizontal-coma,spherical aberration(SA),were significantly elevated in the Verisyse group compared with the ICL group (Z =4.72,4.24,3.12,3.65,2.16;all at P ＜ 0.05).The mean efficacy index was 108.49 ± 16.62 in the V erisyse group and 106.71±15.88 in the ICL group,showing a significant difference (t =0.54,P＞0.05).The mean safety index was 140.56±33.89 in the Verisyse group and 143.34±34.56 in the ICL group,with a significant difference between them (t=0.29,P＞0.05).The mean predictable index was-0.25(-0.97,4.23)in the Verisyse group and 0.98 (-1.44,1.52) in the ICL group,and the difference was significant (Z =-2.68,P ＜ 0.05).Conclusions Both Verisyse PIOL implantation and ICL implantation for high myopia are safe,predictable and effective.The vusial quality and predictability of ICL implantation are much better than Verisyse PIOL implantation.A long-term effect should be observed for the further evaluation.

Objective To explore the role of the SerpinB5 and β-catenin in occurrence and development of the primary hepatocellular carcinoma (HCC). Methods The expressions of SerpinB5 and β-catenin protein and mRNA in carcinoma tissues and paracancerous tissues of 60 patients with primary HCC were detected by immumohistochemistry and real-time quantitative reverse transcriptional polymerase chain reaction (RT-PCR) methods. Results The positive expression rate of SerpinB5 protein and SerpinB5 mRNA in carcinoma tissues were significantly lower than those in paracancerous tissues:25.0%(15/60) vs. 63.3%(38/60) and 1.12 ± 0.43 vs. 5.19 ± 0.39, and there were statistical differences (P<0.01). The positive expression rate of β-catenin protein and β-catenin mRNA in carcinoma tissues were significantly higher than those that in paracancerous tissues: 65.0%(39/60) vs. 31.7%(19/60) and 4.23 ± 0.25 vs. 1.19 ± 0.17, and there was statistical difference (P<0.01). Decreased SerpinB5 expression was associated with higher serumα-fetoprotein level, larger tumor size, poor differentiation, advanced TNM stage, capsule invasion and tumor thrombosis (P < 0.01 or 0.05). Increased β-catenin expression was associated with poor differentiation, advanced TNM stage, capsule invasion and tumor thrombosis (P < 0.01 or < 0.05). The correlation analysis result showed that SerpinB5 had negative correlation withβ-catenin (carcinoma issues:r=-0.346, P=0.001;paracancerous tissues:r=-0.258, P = 0.024). Conclusions The abnormal expression of SerpinB5 and β-catenin may contribute to the progression and biologically malignant behavior of primary HCC, and SerpinB5 and β-catenin exists synergistic effect in the occurrence and development of primary HCC.

[ABSTRACT]OBJECTIVETo explore the occurrence and development of nasal polyps by studying the interleukin 17 and vascular endothelial growth factor expression level in nasal polyp tissues.METHODSNasal polyps in 30 patients with chronic sinusitis (with polyps), ethmoid sinus mucosa in 30 patients with chronic sinusitis (without polyps), inferior turbinate mucosa in 10 patients with nasal septum deviation were collected intraoperatively. IL-17 and VEGF expression were detected using immunohistochemistry SP testing, and correlation between them was analysed. RESULTS1. The expression of IL-17 and VEGF in nasal polyps group is significantly higher than that of the other two groups.There was significant difference between each two groups statistically (P<0.01). 2. The expression of IL-17 was positively correlated with VEGF expression in nasal polyps.CONCLUSION1. The expression of IL-17 and VEGF increased in nasal polyp tissues. 2. The expression of IL-17 and VEGFwas positively correlated in nasal polyps , and both of them take part in the occurrence and development of nasal polyps.

Objective To investigate the association between chronic kidney dysfunction and the complexity of coronary artery disease in elderly patients.Methods A prospective study was conducted on 1380 consecutive patients underwent coronary angiography for the first time in our hospital and with angiographically diagnosed coronary artery disease from January 2011 to June 2012.The complexity of coronary artery disease were classified according to the American College of Cardiology/American Heart Association (ACC/AHA) grading system as types A,B1,B2,and C.Estimated glomerular filtration rate (eGFR) was calculated by the simplified Modification of Diet in Renal Disease(MDRD)equation.Patients were classified into 3 stages according to eGFR as follows:normal renalfunction(n=234,eGFR≥90 ml· min-1 · 1.73 m-2),mild renaldysfunction(n=881,60≤eGFR＜90 ml · min-1 · 1.73 m-2,and moderate or severe renaldysfunction(n=265,eGFR＜60ml · min-1 · 1.73 m-2).Ordinal logistic regression was used to analyze the association between chronic kidney dysfunction and the complexity of coronary artery disease.Results Patients with mild,moderate or severe renal dysfunction were older (F=56.82,P＜0.001),more predominantly female (x2 =66.29,P＜ 0.001) and more likely to have history of hypertension (x2 =17.57,P ＜ 0.001),diabetes (x2=20.97,P＜0.001) and hyperlipidemia (x2=10.48,P 0.005) than those with normal renal function.The percentage of lesions of types B2 or C in moderate or severe renal dysfunction group was higher than that in normal renal function group (x2=175.03,P＜0.001).The ordinal logistic regression showed that age,male,hypertension,diabetes,C-reactive protein and eGFR were independent risk factors for the ACC/AHA lesion classification.Conclusions Age,male,hypertension,diabetes,C-reactive protein and eGFR are independent risk factors for the complexity of coronary artery disease.