Diagnostic efficiency and incremental value of myocardial blood flow quantification by CZT SPECT for patients with coronary artery disease / 中华心血管病杂志

Objective: To investigate the diagnostic efficiency and incremental value of quantitative myocardial blood flow measurements by Cadmium-Zine-Telluride (CZT) single photon emission computed tomography (SPECT) dynamic myocardial perfusion imaging (MPI) in patients with coronary artery disease (CAD) compared with traditional semi-quantitative measurements by MPI. Methods: This is a retrospective, cross-sectional study. We retrospectively analyzed clinical data of patients with suspected or known CAD, who underwent the dynamic MPI quantitative blood flow measurement of CZT SPECT in TEDA International Cardiovascular Hospital from October 2018 to December 2020. Clinical data, semi-quantitative parameters (stress score (SS), rest score (RS) and different score (DS)) and myocardial quantitative blood flow parameters (rest myocardial blood flow (rMBF), stress myocardial blood flow (sMBF) and myocardial flow reserve (MFR)) were analyzed. According to the results of coronary angiography, patients were divided into the stenosis group and the control group with coronary artery stenosis ≥50% or ≥75% as the diagnosis criteria. The differences of quantitative and semi-quantitative parameters between the two groups were compared, and the diagnostic efficacy was compared by receiver operating characteristic(ROC) curve. Results: A total of 98 patients with a mean age of (62.1±8.7) years were included in the study, including 66 males (67%). At the patient level, with the positive standard of coronary artery stenosis≥50%, the left ventricle (LV) stress MBF (LV-sMBF) ((1.36±0.45) ml·min-1·g-1) and LV-MFR (1.45±0.43) of the stenosis group were lower than the LV-sMBF ((2.09±0.64) ml·min-1·g-1) and LV-MFR (2.17±0.54) of control group; summed SS and summed DS were higher than control group (all P<0.05). With the positive standard of coronary artery stenosis ≥75%, the LV-sMBF ((1.19±0.34) ml·min-1·g-1) and LV-MFR (1.34±0.35) of stenosis group were lower than the LV-sMBF ((1.94±0.63) ml·min-1·g-1) and MFR (2.00±0.58) of control group; all semi-quantitative parameters were higher than control group (all P<0.05). At the vascular level, with coronary artery stenosis ≥50% as the diagnosis criteria, the sMBF ((1.26±0.49) ml·min-1·g-1) and MFR (1.35±0.46) of stenosis group were lower than the sMBF ((1.95±0.70) ml·min-1·g-1) and MFR (2.05±0.65) of control group; SS and DS were higher than control group (all P<0.05). With coronary artery stenosis≥75% as the diagnosis criteria, the sMBF ((1.12±0.41) ml·min-1·g-1) and MFR (1.25±0.38) of stenosis group were lower than the sMBF ((1.84±0.70) ml·min-1·g-1) and MFR (1.93±0.66) of control group; all semi-quantitative parameters were higher than control group (all P<0.05). With coronary artery stenosis≥50% as the diagnosis criteria and CAG as the reference standard, the AUC and 95%CI of myocardial quantitative blood flow parameters indicated by ROC curve for diagnosis of CAD were 0.830 (0.783-0.877). The sensitivity (86.1% vs. 61.5%), specificity (82.6% vs. 73.8%), positive predictive value (77.8% vs. 62.5%), negative predictive value (89.3% vs. 73.0%) and accuracy (84.0% vs. 68.7%) were all higher than the semi-quantitative parameters (all P<0.05). With coronary artery stenosis≥75% as the diagnosis criteria, the AUC and 95%CI of myocardial quantitative blood flow parameters indicated by ROC curve for diagnosis of CAD were 0.832(0.785-0.879). The sensitivity (89.2% vs. 67.6%), negative predictive value (95.5% vs. 86.2%) and accuracy (80.6% vs. 68.0%) were all higher than semi-quantitative parameters (all P<0.05). Conclusion: Compared with traditional SPECT MPI derived semi-quantitative parameters, diagnostic efficacy for CAD is higher using CZT SPECT quantitative myocardial blood flow parameters, this strategy thus has additional diagnostic benefits and incremental value on the diagnosis of CAD.

Mechanism of Cervi Cornu Pantotrichum in Treatment of Osteoarthritis Based on Network Pharmacology / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Cervi Cornu Pantotrichumin in the treatment of osteoarthritis by network pharmacology. MethodThe active ingredients and the corresponding targets of Cervi Cornu Pantotrichumin were screened out by a Bioinformatics Analysis Tool of Molecular mechANism of Traditional Chinese Medicine （BATMAN-TCM）. The targets related to osteoarthritis were obtained through GeneCards and Online Mendelian Inheritance in Man （OMIM）. The targets corresponding to the active ingredients and those related to osteoarthritis were intersected to reveal the common targets， and STRING was adopted to build a protein-protein interaction （PPI） network. DAVID was used for gene ontology （GO） annotation and Kyoto encyclopedia of genes and genomes （KEGG） pathway enrichment on the anti-osteoarthritis targets of Cervi Cornu Pantotrichumin， and R x64 3.6.3 was employed to produce the advanced bubble charts of GO terms and KEGG pathways. Cytoscape 3.7.2 was used to establish the “Chinese medicinal herb-active ingredient-target-signaling pathway” network. In vitro experiments were performed to detect the viability of RAW 264.7 cells exposed to oxidative stress and the tumor necrosis factor （TNF）-α level in RAW 264.7 cells with inflammation under the treatment by Cervi Cornu Pantotrichumin. ResultA total of 20 active ingredients of Cervi Cornu Pantotrichum were obtained， of which ceramide， 6'-O-β-D-glucosylgentiopicroside， cerebroside， oleuropein， sphingomyelin， and cholesterol ferulate did not meet the screening conditions. Therefore， a total of 14 active ingredients were finally screened out， and 303 and 3 093 targets of active ingredients and osteoarthritis were respectively obtained. The two target sets were taken to intersect， which revealed 92 common targets. GO annotation and KEGG pathway enrichment showed that the targets were mainly involved in redox process， positive regulation of RNA polymerase Ⅱ promoter transcription， inflammatory response， protein synthesis， osteoclast differentiation， TNF signaling pathway， signaling pathways in cancer， mammalian target of rapamycin （mTOR） signaling pathway， mitogen-activated protein kinase （MAPK） signaling pathway， and cyclic adenosine monophosphate （cAMP） signaling pathway. The results of in vitro experiments showed that a certain concentration of protein in Cervi Cornu Pantotrichum significantly increased the viability of RAW 264.7 cells exposed to H2O2-induced oxidative damage （P<0.05， P<0.01） and reduced the level of TNF-α in the RAW 264.7 cells experiencing lipopolysaccharide （LPS）-induced inflammation （P<0.05）. ConclusionBased on the network pharmacology method， the mechanism of the multi-component， multi-target and multi-pathway treatment of OA by antler antler was explained， and the anti-inflammatory and antioxidant activities of antler antler were confirmed， which provided theoretical guidance and scientific basis for further research on the treatment of OA by antler antler.

Study on antidepressant effect of total alkaloids of Fibraurea recisa / 中国中药杂志

This study aimed to investigate the antidepressant effects of total alkaloids of Fibraurea recisa in HT22 cells damaged by corticosterone (CORT) in vitro and in a mouse model of chronic unpredictable mild stress (CUMS) as well as the underlying mechanisms.In cellular experiments,the viability of CORT-damaged HT22 cells was detected using cell counting kit-8 (CCK-8),and the cell apoptosis was detected by Hoechst 33258 staining.In animal experiments,C57BL/6N mice were randomly divided into the control group,model group,low (100 mg·kg~(-1)),medium (200 mg·kg~(-1)) and high (400 mg·kg~(-1))-dose of total alkaloids of F.recisa groups,and positive control group.After 21 days of CUMS exposure,their depressive behaviors were observed in behavioral and Morris water maze tests.The serum levels of 5-hydroxytryptamine (5-HT),dopamine (DA),and norepinephrine (NE) were assessed by ELISA.The expression levels of apoptosis-related proteins Bcl-2,Bax and cleaved caspase-3 in HT22 cells and mouse hippocampus were detected by Western blot.The results suggested that total alkaloids of F.recisa alleviated the damage of HT22 cells induced by CORT in a dose-dependent manner.The Hoechst 33258 staining uncovered that total alkaloids of F.recisa better reduced the blue spots and inhibited cell apoptosis.The results of animal experiments showed that total alkaloids of F.recisa significantly improved the depression-like behaviors of mice and increased the serum levels of 5-HT,DA and NE as compared with those in the model group.The Western blot assays revealed a significant up-regulation of Bcl-2 protein expression,but an obvious reduction in Bax and cleaved caspase-3protein expression in the total alkaloids of F.recisa group.In conclusion,total alkaloids of F.recisa inhibited depression possibly by regulating the apoptosis-related protein expression or elevating the monoamine neurotransmitter levels in the brain.

Related risk factors of PET/CT detected coronary microvascular disease in patients with chest pain and no obstructive coronary artery disease / 中华心血管病杂志

Objective: To explore the related factors of the coronary microvascular disease (CMD) diagnosed with positron emission tomography(PET)/CT in patients with chest pain and without obstructive coronary artery disease (NOCA). Methods: This study was a single-center retrospective cross-sectional study. Consecutive patients with chest pain and NOCA on coronary angiography, who underwent PET/CT quantitative myocardial blood flow measurements at TEDA International Cardiovascular Hospital from August 2018 to January 2019, were enrolled for this study. The diagnostic criteria for NOCA was the absence of coronary artery diameter stenosis ≥50% on coronary angiography. Clinical data, global left ventricular myocardial blood flow on stress and rest, and the coronary flow reserve (CFR) were analyzed. Patients were divided into two groups according to CFR. Patients with CFR<2 were defined as CMD group, and the rest were classified as control group. Pearson correlation analysis and Logistics regression analysis were used for exploring the risk factors of the CMD. Results: A total of 66 patients, with an mean age of (56.7±9.6) years, were included in the study, including 41 females (62%). There were 20 patients with CMD (30%). Body mass index (BMI) was significantly higher in CND group than in control group ((28.1±3.6) kg/m2 vs. (25.6±3.5) kg/m2, P=0.01). Total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) were also significantly higher in CMD group than in control group ((4.89±1.03) mmol/L vs. (4.30±1.02) mmol/L and (3.23±0.81) mmol/L vs. (2.71±0.95) mmol/L respectively, P=0.038). Pearson correlation analysis showed that CFR was moderately correlated with BMI (r=-0.45, P<0.001), and was weakly correlated with TC and LDL-C (r=-0.271 and r=-0.280, respectively, P<0.05). Multivariate logistic regression analysis showed that BMI (the risk of CMD increased by 1.528 times for every 5 kg/m2 increase in BMI, 95%CI 1.083-5.897, P<0.05) was an independent risk factor of CMD after adjusted by gender, hypertension, diabetic mellites and LDL-C. Conclusion: For patients with NOCA and chest pain, high BMI is independent risk factor of CMD diagnosed by PET/CT.

Study advances in regulation effect of Tripterygium wilfordii and its extracts on innate immune system in rheumatoid arthritis cases / 中国中药杂志

Rheumatoid arthritis( RA) is an autoimmune disease characterized by chronic and aggressive polyarthritis. The innate immunity mechanism plays a key role in the pathogenesis of RA. Tripterygium wilfordii and its extracts have regulatory effects on innate immune cells including macrophages,dendritic cells,neutrophils,mast cells,NK cells,NKT cells,etc.,as well as a variety of innate immune molecules including cytokines,adhesion molecules,patterns recognition receptor( PRR) and the complement molecules,showing a regulatory effect in the pathogenesis of RA innate immunity. In this paper,the recent domestic and foreign researches on the pathogenesis of RA with innate immunity involved were reviewed and the research status of T. wilfordii and its extracts on the regulation of innate immunity involved in RA was summarized.

Advance in studies on pharmacokinetics of baicalin / 中国中药杂志

Scutellariae baicalensis is one of the most important traditional Chinese medicinal herbs, mainly distributed in Shandong and Hebei provinces. It has significant pharmacological effects such as antimicrobial activity, anti-inflammatory and antioxidation. Baicalin is one of its main effective components. However, baicalin's low bioavailability has restricted its clinical application. In recent decades, extensive studies have been carried out on the metabolism of baicalin at home and abroad. In order to provide scientific references for baicalin's further studies, this paper would not only review the advances in pharmacokinetics research of baicalin and Chinese herbal preparations containing baicalin, but also make a summary on research status of baicalin.

miR-146b facilitates the differentiation of induced pluripotent stem cells into neuron-like cells / 中国组织工程研究

BACKGROUND: To facilitate the neuronal differentiation of induced pluripotent stem cells (iPSCs) is of great benefit for the repair of nerve injury and nerve regeneration. OBJECTIVE: To investigate the expression of miR-146b in iPSCs differentiation, and its effects on the neural differentiation of iPSCs. METHODS: After 7-day neural induction of mouse iPSCs, the expressions of miR-146b and pluripotent stem cell markers were detected by real-time PCR. Then, we generated miR-146b overexpressing iPSCs followed by the neural induction. The expression of stem cell and neuroectoderm markers were examined by cell immunofluorescence and real-time PCR respectively. Meanwhile, the expression of Tuj-1, a nerve cell marker, was also detected. RESULTS AND CONCLUSION: Compared with the normal iPSCs, the expressions of pluripotent stem cell markers Nanog, Oct4 and Sox2 were significantly down-regulated in iPSCs undergoing 7-day neural induction, while the level of miR-146b was increased obviously. In miR-146b-overexpressing iPSCs, we found that the expression of Oct4 was substantially decreased, while there were no statistical changes in the expression of Nanog and Sox2. Meanwhile, during the neural differentiation, miR-146b overexpression significantly down-regulated the expression of Nanog, Oct4 and Sox2, and up-regulated the expression of neuroectoderm markers, Nestin, Sox1 and Pax6. After 18 days of neural induction, the miR-146b overexpression significantly increased the number of Tuj-1-positive cells. Taken together, miR-146b plays a vital role in facilitating the differentiation of iPSCs into neuron-like cells.

The application of poly (2-ethyl-2-oxazoline) in drug delivery system / 药学学报

Poly(2-ethyl-2-oxazoline) (PEOz), a hydrophilic long-chain polymer synthesized by living cationic ring-opening isomerization polymerization of 2-ethyl-2-oxazoline, has the characteristics of low toxicity, biocompatibility, flexible chain, and modified expediently. PEOz is a potential substitute of polyethylene glycol (PEG) to render the ability of long-circulation, enhance cellular uptake and endosomal escape behaviors to PEOz modified drug delivery system. In this review, we summarized recent literature for the research progress of physicochemical properties, synthetic methods and the application of PEOz in drug delivery system.

Determination of aflatoxins in fruit traditional Chinese medicines by rapid-resolution liquid chromatography tandem mass spectrometry / 中国药学杂志

OBJECTIVE: To develope a method for determination of four aflatoxins (B1, B2, G1, G2) in fruit traditional Chinese medicines by rapid-resolution liquid chromatography coupled with electrospray ionization triple quadruple tandem mass spectrometry. METHODS: After being extracted with 70% methanol and purified by immunoaffinity columns, the four aflatoxins were separated on a Shim-pack XR-ODS column using gradient elution with the mobile phase of CH3CN-CH3OH-H2O mixtures monitored under MRM mode in ESI positive scan. RESULTS: There was a good linear relationship over the range of 0.102 1-10.21 ng · mL-1 for AFB1, 0.061 2-7.65 ng · mL-1 for AFB2, 0.193-9.65 ng · mL-1 for AFG1, 0.121-7.55 ng · mL-1 for AFG2 (r > 0.999 8), respectively. The average recoveries ranged from 77.0%-102.4%. CONCLUSION: This method provides a simple and feasible way to determine aflatoxins.

The relationship between perfusion defects on myocardial SPECT and stenotic severity on CT coronary angiography / 中华核医学杂志

ObjectiveTo evaluate the diagnostic performance of CTCA in predicting myocardial perfusion defects through comparative analysis between MPI defects and severity of coronary stenosis on CTCA.MethodsFour hundred and seventy-eight patients who underwent CTCA and 99Tcm-MIBI MPI in the same period were analyzed retrospectively.According to the severity of coronary stenosis judged by visual evaluation of the vessel diameter,the patients were divided into five groups:no stenosis,mild stenosis,moderate stenosis,severe stenosis and total occlusion.MPI results were classified as negative or positive for perfusion defects,and the prevalence of perfusion defects in every group was calculated per-patient and per-vessel basis.The cut-off of stenotic severity for predicting myocardial perfusion defects was designated as 50％ or 75％,with MPI as standard reference.True positive,true negative,false positive and false negative statistics were thus determined separately on patient and vessel basis.The diagnostic performance for CTCA were calculated and compared.Pearson Chi-square and its partition tests or Fisher exact test were used to compare ordinal variables.ResultsFifty-eight patients showed myocardial perfusion defects.Either by patientbased or vessel-based analysis,the prevalence of myocardial perfusion defects showed an increasing trend with greater coronary artery stenosis in each group,and there were statistical differences among them (x2 =116.62 and 483.83,both P ＜ 0.05).On patient-based analysis,sensitivity ( SN),specificity ( SP),positive predictive value( PPV),negative predictive value(NPV) and accuracy (AC) for CTCA predicting myocardial perfusion defects were 62.1 ％ ( 36/58 ) and 34.5％ ( 20/58 ) (x2 =8.84,P ＜ 0.05 ),84.5％(355/420) and 97.1％ (408/420) (x2 =40.16,P ＜0.05),35.6％ (30/101) and 62.5％ (20/32) (x2 =7.19,P＜0.05),94.2％ (355/377) and 91.5％ (408/446) (x2 =2.18,P ＞0.05),81.8％ (391/478)and 89.5 ％ (428/478) (x2 =11.66,P ＜ 0.05 ) when the cutoff was set to 50％ and 75％,respectively.On vessel-based analysis,the SN,SP,PPV,NPV and AC for CTCA predicting myocardial perfusion defects were 58.8％ (40/68) and 30.9％ (21/68) (x2 =10.73,P ＜ 0.05),95.9％ (1768/1844) and 99.0％ (1826/1844) (x2 =36.72,P ＜ 0.05 ),34.5％ (40/116) and 53.8％ (21/39) (x2 =4.59,P ＜0.05 ),98.4％ (1768/1796) and 97.5％ ( 1826/1873 ) (x2 =4.14,P ＜ 0.05 ),94.6％ ( 1808/1912 ) and 96.6％ ( 1847/1912 ) (x2 =10.31,P ＜ 0.05 ),respectively.ConclusionsThe prevalence of myocardial perfusion defects correlates positively with the severity of coronary stenosis seen on CTCA.CTCA may predict perfusion defects with high SP and NPV.However,the PPV of CTCA in predicting myocardial perfusion defects is poor when the stenosis cut-off is set at 50％.It is significantly improved when the cutoff value is set at 75 ％.

Pacemaker/ICD lead extraction with the excimer laser sheath: a single center experience / 中华心血管病杂志

<p><b>OBJECTIVE</b>The aim of this study is to investigate the safety and effectiveness of Excimer laser-assisted lead extraction.</p><p><b>METHODS</b>Consecutive lead extraction in 16 patients using the excimer laser sheath from Veterans Administration Medical Center, University of Minnesota between March 2008 and December 2009 were analyzed retrospectively. The clinical characteristics including basic disease, cardiac function, indication and reason for lead removal, lead characteristic, and complication were summarized.</p><p><b>RESULTS</b>Thirty one leads (laser sheaths were used in 25 leads) in 16 patients [mean age (67.4 ± 9.1) years, range 54 - 82 years] were extracted. All leads were extracted successfully without complication. Most patients had severe heart diseases with left ventricular ejection fraction of 20% - 60% (mean 36.4% ± 13.4%). Mean implant time was (73.9 ± 62.9) months (range 3 - 251 months). Nine patients had Class I indications for lead extraction, 7 patients for Class II indications. Atrial lead fractured in one patient during lead extraction, the remaining part of the lead was taken out from femoral vein by a snare-catheter apparatus. Another patient developed CS dissection and small pericardium perfusion during LV lead implantation (upgrade to BiV-ICD) post successful lead extraction and patient discharged without complication.</p><p><b>CONCLUSION</b>Our single center experience suggests that Excimer Laser Sheath Lead Removal System is safe and effective in extracting pacemaker and ICD leads.</p>

Establishment of a colorectal cancer cell line with PRL-3 and CDH22 gene knock-down by lentivirus-mediated RNA interference / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a colorectal cancer cell line with stable PRL-3 and CDH22 gene knock-down for investigating the role of PRL-3 and CDH22 genes in the carcinogenesis and metastasis of colorectal cancer.</p><p><b>METHODS</b>A recombinant lentiviral vector targeting CDH22 gene was obtained using the pENTRTM/U6 construct and pLenti6/BLOCK-iT (TM)-DEST vector. The recombinant lentivirus was harvested from 293FT cells cotransfected with the optimized ViraPower(TM) Packaging Mix and the pLenti6/BLOCK-iT(TM)-DEST expression construct. SW480/PRL-3- cells were infected with the recombinant lentivirus targeting CDH22, and SW480 cells with stable PRL-3 and CDH22 knock-down were screened by blasticidin selection. PRL-3 expression in the cells was determined by real-time RT-PCR. RESULTS The titer of the lentivirus for the second infection was 8 x 10(5) U/ml. Seventeen positive clones were selected, among which the Clone 1 exhibited substantially down-regulated CDH22 and PRL-3 mRNA expressions.</p><p><b>CONCLUSIONS</b>A human colorectal cancer cell line with stable PRL-3 and CDH22 gene knock-down has been established.</p>

Comparison of full-field digital mammography and magnetic resonance imaging for breast disease diagnosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the effectiveness of full-field digital mammography (FFDM) and magnetic resonance imaging (MRI) in the detection and diagnosis of breast diseases.</p><p><b>METHODS</b>Forty-one patients with 47 breast lesions (20 malignant and 27 benign lesions) underwent preoperative FFDM and MRI, using the pulse sequences including T1WI, T2WI, T2WI/SPIR diffusion-weighted imaging (DWl), and 3D dynamic contrast-enhanced MRI.</p><p><b>RESULTS</b>The imaging and pathological findings were compared, and the detection rates of the lesions by FFDM and MRI were 97.87% (46/47) and 74.46% (35/47), respectively (P<0.01). The sensitivity of FFDM and MRI was 70.00% (14/20) and 80.00% (16/20) (P>0.05), with specificity of 62.96%(17/27) and 88.89%(24/27) (P<0.05), respectively.</p><p><b>CONCLUSION</b>MRI is superior to FFDM in the detection and diagnosis of breast diseases.</p>

Indications and effect of the intra-aortic balloon pump in 38 patients during the cardiac surgery / 中南大学学报(医学版)

OBJECTIVE@#To review the indications and effect of the intra-aortic balloon pump (IABP) in 38 patients during the cardiac surgery.@*METHODS@#From June 2004 to August 2007, 38 patients were performed IABP in the cardiac surgery, including 28 coronary artery bypass grafting (CABG) and 10 valve replacements. The intra-operation prophylactic use of IABP was in 5 cases, the use of IABP for weaning from cardiopulmonary bypass (CPB) during the cardiac surgery was in 22 cases, and the rest use of IABP was after the operation in the intensive care unit. Low molecular weight heparin was used for IABP anticoagulation.@*RESULTS@#Thirteen died out of the 38 IABP patients. The total IABP success rate was 66%.The IABP success rate in CABG was 82%, and 20% in the valve replacement. The patients with intra-operation prophylactic use of IABP all recovered, and 16 recovered out of the 22 patients with IABP uses for weaning from CPB, and 6 recovered out of the 11 patients with post-operation IABP application. No severe complications related to IABP occurred.@*CONCLUSION@#IABP has better effect in CABG patients than in valve replacements. Effect of the prophylactic use of IABP is better than both the IABP use for weaning from CPB or the post-operation IABP application. For IABP anticoagulation, low molecular weight heparin is the better alternative than routine heparin, and the proper ACT is controlled at 150-170 s.

Controlled clinical trials on the pathological expressions of VEGF in patients with active ankylosing spondylitis / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the pathological expression and significance of VEGF in patients with active ankylosing spondylitis.</p><p><b>METHODS</b>The expression of VEGF in the synovial tissues of cacroiliac joint of patients with active AS was detected by using in situ hybridization and the results were compared with those in the patients with pelvic fracture using image analysis system.</p><p><b>RESULTS</b>The positive expressions of VEGF in the synovial tissues of cacroiliac joint of patients with active AS were stronger than those in the control group (P<0.01).</p><p><b>CONCLUSION</b>VEGF are important factors in patients with active AS. They are tightly correlated with the process of osteoclasia and pathological new bone formation in the cacroiliac joint of patients with active AS. If we can reduce the expressions of VEGF in the patients with active AS, the process of osteoclasia and pathological new bone formation will be interrupted and this provides a new strategy for the treatment of ankylosing spondylitis.</p>

Construction of a lentiviral vector for RNA interference of PRL-3 gene and its stable expression in SW480 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lentiviral vector for RNA interference (RNAi) of PRL-3 gene and establish a human colon carcinoma cell line with PRL-3 gene knock-down.</p><p><b>METHODS</b>The plasmids were constructed expressing two different short hairpin RNAs (shRNA) targeting PRL-3 gene under control by the U6 promoter by lentiviral vector. An entry clone was generated in the pENTR/U6 vector. After identification with sequencing, a recombinant lentiviral vector was obtained using the pENTR/U6 construct and pLenti6/BLOCK-iT TM-DEST vector. The recombinant lentivirus was harvested from 293FT cells co-transfected with optimized ViraPower Packaging Mix and the pLenti6/BLOCK-iT -DEST expression construct. SW480 cells were infected with the recombinant lentivirus and the cells with stable PRL-3 gene knock-down were screened by blasticidin selection. PRL-3 expression was detected using real-time reverse transcription-polymerase chain reaction and Western blotting.</p><p><b>RESULTS</b>A recombinant lentiviral vector expressing shRNAs targeting PRL-3 gene was successfully established and confirmed by DNA sequencing. The recombinant lentivirus were harvested from 293FT cells with a viral titer of 6 x 10(5) pfu/L. Twelve clones of SW480 cells infected with the recombinant lentivirus were selected, and the Clone 1 exhibited significant knock-down of PRL-3 protein expression (72.9%).</p><p><b>CONCLUSION</b>The successful establishment of human colon carcinoma cell line with PRL-3 gene knock-down provide a basis for investigation of the role of PRL-3 gene in the metastasis of human colorectal carcinoma.</p>

Construction of a lentiviral vector for RNA interference of human CDH22 gene and its gene silencing effect in SW480 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lentiviral expression vector for RNA interference of human CDH22 gene, and assess its gene silencing effect in colorectal cancer cells to provide a basis for investigating the role of CDH22 gene in the signaling pathway involved in human colorectal carcinoma metastasis.</p><p><b>METHODS</b>Human CDH22 gene short hairpin RNA (shRNA) sequence was designed using a software available on-line. After synthesis and annealing, the double-stranded oligonucleotides (dsOligoe) were cloned into the pENTR(TM)/U6 plasmid followed by sequence analysis. A positive clone was subcloned into pLenti6/BLOCK-iT(TM)-DEST vector and transformed into stb13 competent cells, with also verification by sequencing. The recombinant lentivirus was harvested from 293FT cells contransfected with the positive recombined plasmid and lentiviral packing materials. SW480 cells were infected with the recombinant lentivirus and the cells with stable CDH22 knock-down were screened by blasticidin selection. CDH22 expression in the cells was determined by real-time reverse transcription-polymerase chain reaction.</p><p><b>RESULTS</b>A recombinant lentiviral vector expressing shRNAs against CDH22 gene was obtained and confirmed by DNA sequencing. Fifteen clones of SW480 cells infected with the recombinant lentivirus were selected, and clone 11 exhibited substantial knock-down of CDH22 mRNA expression.</p><p><b>CONCLUSION</b>The lentiviral shRNA expression vector targeting human CDH22 gene capable of stable CDH22 gene knock-down in SW480 cells has been successfully constructed, which provides a basis for further study of the relationship between human colorectal carcinoma and CDH22 gene.</p>

Expression of the zinc finger transcriptional factor Snail in colorectal carcinoma and its significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To study expression of the zinc finger transcriptional factor Snail in colorectal carcinoma and its significance.</p><p><b>METHODS</b>Expressions of Snail in colorectal carcinoma SW480 and SW620 cells were assayed by immunocytochemistry and immunofluorescent cytochemistry. The paraffin-embedded specimens from 68 cases of colorectal carcinoma and its corresponding adjacent tissues, 33 cases of adenoma and 35 cases of metastatic lymph nodes were also examined for Snail expressions using immunohistochemistry.</p><p><b>RESULTS</b>Snail protein was located mainly in the cell nucleus of SW480 and SW620 cells. The expressions of PRL-3 protein in the specimens of colorectal carcinoma, normal mucosa, adenoma and metastatic lymph nodes were significantly different (Chi(2)=92.852, P=0.000). In the adenoma tissues, the expression was significantly higher than that in normal mucosa (Z=-2.902, P=0.004), the metastatic lymphnodes had significantly higher expressions than the primary colorectal carcinomas (Z=-4.951, P=0.000), which, in turn, showed significantly higher expression than the adenoma tissues (Z=-3.572, P=0.000). Significant correlation of Snail expression was found to the progression and metastasis of colorectal carcinoma (Z=-2.043, P=0.041).</p><p><b>CONCLUSION</b>The expression of Snail is significantly correlated to genesis, progression and metastasis of colorectal carcinoma.</p>

Cloning of human PRL-3 gene and construction of its prokaryotic expression vector / 南方医科大学学报

<p><b>OBJECTIVE</b>To obtain the entire coding sequence of human PRL-3 gene and construct its prokaryotic expression vector.</p><p><b>METHODS</b>With total RNA extracted from the human colorectal carcinoma cell line SW480 as the template, PRL-3 gene was amplified by RT-PCR with primers designed according to the published sequence of GenBank, and the product was inserted into pGEM-T Easy vector. The recombinant plasmid pGEM-T-PRL-3 was identified by restriction endonuclease analysis and DNA sequencing. After digestion with restriction endonuclease, PRL-3 gene was cloned into the multicloning sites of the prokaryotic expression vector pGEX-4T-1.</p><p><b>RESULTS AND CONCLUSION</b>The entire coding region of human PRL-3 gene was cloned, and the recombinant pGEX-4T-1-PRL-3 vector was successfully constructed and expressed, which may provide the basis for further study of the relationship between human colorectal carcinoma and PRL-3 gene.</p>

Preliminary study of PRL-3 gene promoter binding sites of Snail in SW480 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify the region in PRL-3 gene promoter where the transcriptional factor Snail can bind.</p><p><b>METHODS</b>PRL-3 promoter and the possible binding sites of the transcription factors were analyzed by bioinformatical methods. Chromatin immunoprecipitation and PCR were performed using the antibody specific for Snail to verify the binding of Snail to PRL-3 promoter.</p><p><b>RESULTS</b>According to the prediction by TRED, a promoter prediction software, PRL-3 gene promoter was located between -700 bp to 299 bp of PRL-3 gene. Many possible transcription factor binding sites such as for Snail, n-MYC, ARNT, E74A, NF-kappaB, NRF-2 and AML-1 were predicted by Consite, a promoter analysis web system. Interestingly, a 5'-CACCTG-3' core sequence and other related sequences of Snail binding sites were found in the promoter region of PRL-3 genes by Consite software. Two regions in PRL-3 promoter were validated to allow binding of Snail by chromatin immunoprecipitation analysis of SW480 cells.</p><p><b>CONCLUSIONS</b>Snail regulates the activity of PRL-3 gene by binding to the promoter of PRL-3 gene in SW480 cells.</p>