Clinical value of chemiluminescence method for detection of antinuclear antibody profiles

BACKGROUND Antinuclear antibodies(ANAs)are crucial in diagnosing autoimmune diseases,mainly systemic lupus erythematosus(SLE).This study aimed to compare the performance of chemiluminescence assay(CLIA)and line immunoassay(LIA)in detecting ANAs in patients with autoimmune diseases,evaluate their diagnostic accuracy for SLE,and develop a novel diagnostic model using CLIA-detected antibodies for SLE.Specimens from patients with autoimmune diseases and physical examination specimens were collected to parallel detect specific antibodies.Individual antibodies'diagnostic performance and a model combining multiple antibodies were assessed.The findings provide valuable insights into improving the diagnosis of SLE through innovative approaches.AIM To compare the performance of CLIA and LIA in detecting ANAs in patients with autoimmune diseases,assess their accuracy for SLE,and develop a novel diagnostic model using CLIA-detected antibodies for SLE.METHODS Specimens have been obtained from 270 patients with clinically diagnosed autoimmune disorders,as well as 130 physical examination specimens.After that,parallel detection of anti-double-stranded DNA(dsDNA)antibody,anti-histone(Histone)antibody,anti-nucleosome(Nuc)antibody,anti-Smith(Sm)antibody,anti-ribosomal P protein(Rib-P)antibody,anti-sicca syndrome A(Ro60)antibody,anti-sicca syndrome A(Ro52)antibody,anti-sicca syndrome(SSB)antibody,anticentromere protein B(Cenp-B)antibody,anti-DNA topoisomerase 1(Scl-70)antibody,anti-histidyl tRNA synthetase(Jo-1)antibody,and anti-mitochondrial M2(AMA-M2)antibody was performed using CLIA and LIA.The detection rates,compliance rates,and diagnostic performance for SLE were compared between the two methodologies,followed by developing a novel diagnostic model for SLE.RESULTS CLIA and LIA exhibited essentially comparable detection rates for anti-dsDNA antibody,anti-Histone antibody,anti-Nuc antibody,anti-Sm antibody,anti-Rib-P antibody,anti-Ro60 antibody,anti-Ro52 antibody,anti-SSB antibody,anti-Cenp-B antibody,anti-DNAScl-70 antibody,anti-Jo-1 antibody and anti-AMA-M2 antibody(P>0.05).The two methods displayed identical results for the detection of anti-dsDNA antibody,anti-Histone antibody,anti-Nuc antibody,anti-Sm antibody,anti-Ro60 antibody,anti-Ro52 antibody,anti-SSB antibody,anti-Cenp-B antibody,anti-Scl-70 antibody,and anti-AMA-M2 antibody(Kappa>0.7,P<0.05),but showed a moderate agreement for the detection of anti-Rib-P antibody and anti-Jo-1 antibody(Kappa=0.671 and 0.665;P<0.05).In addition,the diagnostic performance of these antibodies detected by both methods was similar for SLE.The diagnostic model's area under the curve values,sensitivity,and specificity,including an anti-dsDNA antibody and an anti-Ro60 antibody detected by CLIA,were 0.997,0.962,and 0.978,respectively.These values were higher than the diagnostic performance of individual antibodies.CONCLUSION CLIA and LIA demonstrated excellent overall consistency in detecting ANA profiles.A diagnostic model based on CLIA-detected antibodies can successfully contribute to developing a novel technique for detecting SLE.

Jejunal sarcomatoid carcinoma:A case report and review of literature

BACKGROUND Sarcomatoid carcinoma(SCA)of the jejunum is a rare and aggressive neoplasm affecting the smooth muscle cells of the jejunum.This study presents a recent case of jejunal SCA,detailing its diagnosis and treatment,thereby providing a reference for clinical practice.CASE SUMMARY A 65-year-old male presented to Yichang Central People's Hospital with a chief complaint of hemorrhoids.A computed tomography(CT)scan incidentally revealed multiple abnormal signals in the liver.Subsequent positron emission tomography/CT at Wuhan Union Hospital indicated malignant tumor progression,with a primary duodenal tumor and multiple metastases in the upper left abdomen.Intraoperatively,a large tumor was identified on the omentum.Histopathological and immunohistochemical analyses of the resected specimen confirmed the diagnosis of jejunal SCA.The patient received a combination therapy of sintilimab,nanoparticle albumin-bound paclitaxel,and anlotinib.Follow-up imaging demonstrated significant reduction of hepatic and peritoneal lesions.The patient has remained stable for over one year postoperatively.CONCLUSION This case suggests that chemotherapy,immunotherapy,plus targeted therapy may represent an optimal treatment for intestinal SCA,meriting further investigation.

Analysis of cancer-specific survival in patients with metastatic colorectal cancer: A evidence-based medicine study

BACKGROUND Metastatic colorectal cancer(mCRC)is a common malignancy whose treatment has been a clinical challenge.Cancer-specific survival(CSS)plays a crucial role in assessing patient prognosis and treatment outcomes.However,there is still li-mited research on the factors affecting CSS in mCRC patients and their corre-lation.AIM To predict CSS,we developed a new nomogram model and risk grading system to classify risk levels in patients with mCRC.METHODS Data were extracted from the United States Surveillance,Epidemiology,and End Results database from 2018 to 2023.All eligible patients were randomly divided into a training cohort and a validation cohort.The Cox proportional hazards model was used to investigate the independent risk factors for CSS.A new nomogram model was developed to predict CSS and was evaluated through internal and external validation.RESULTS A multivariate Cox proportional risk model was used to identify independent risk factors for CSS.Then,new CSS columns were developed based on these factors.The consistency index(C-index)of the histogram was 0.718(95%CI:0.712-0.725),and that of the validation cohort was 0.722(95%CI:0.711-0.732),indicating good discrimination ability and better performance than tumor-node-metastasis staging(C-index:0.712-0.732).For the training set,0.533,95%CI:0.525-0.540;for the verification set,0.524,95%CI:0.513-0.535.The calibration map and clinical decision curve showed good agreement and good potential clinical validity.The risk grading system divided all patients into three groups,and the Kaplan-Meier curve showed good stratification and differentiation of CSS between different groups.The median CSS times in the low-risk,medium-risk,and high-risk groups were 36 months(95%CI:34.987-37.013),18 months(95%CI:17.273-18.727),and 5 months(95%CI:4.503-5.497),respectively.CONCLUSION Our study developed a new nomogram model to predict CSS in patients with synchronous mCRC.In addition,the risk-grading system helps to accurately assess patient prognosis and guide treatment.

Machine learning prediction model for gray-level co-occurrence matrix features of synchronous liver metastasis in colorectal cancer

BACKGROUND Synchronous liver metastasis(SLM)is a significant contributor to morbidity in colorectal cancer(CRC).There are no effective predictive device integration algorithms to predict adverse SLM events during the diagnosis of CRC.AIM To explore the risk factors for SLM in CRC and construct a visual prediction model based on gray-level co-occurrence matrix(GLCM)features collected from magnetic resonance imaging(MRI).METHODS Our study retrospectively enrolled 392 patients with CRC from Yichang Central People’s Hospital from January 2015 to May 2023.Patients were randomly divided into a training and validation group(3:7).The clinical parameters and GLCM features extracted from MRI were included as candidate variables.The prediction model was constructed using a generalized linear regression model,random forest model(RFM),and artificial neural network model.Receiver operating characteristic curves and decision curves were used to evaluate the prediction model.RESULTS Among the 392 patients,48 had SLM(12.24%).We obtained fourteen GLCM imaging data for variable screening of SLM prediction models.Inverse difference,mean sum,sum entropy,sum variance,sum of squares,energy,and difference variance were listed as candidate variables,and the prediction efficiency(area under the curve)of the subsequent RFM in the training set and internal validation set was 0.917[95%confidence interval(95%CI):0.866-0.968]and 0.09(95%CI:0.858-0.960),respectively.CONCLUSION A predictive model combining GLCM image features with machine learning can predict SLM in CRC.This model can assist clinicians in making timely and personalized clinical decisions.

Cardiovascular Risk of Opioids: A Real-World Study Based on FAERS

Objective:This research utilizes the FAERS for data mining to identify heart-related side effects caused by opioids,ensuring the safe use of these medications.Methods:Data from 79 quarters(Q12004 to Q32023)involving adverse event(AE)reports for opioids like morphine and oxycodone was reviewed.We applied the MedDRA system to categorize events and used statistical tools,ROR and BCPNN,for signal detection.These findings were cross-checked with drug labels and SIDER 4.1 for accuracy.Identified risks were then categorized by severity using DME and IME classifications.Results:Analysis of adverse events(AEs)for the five examined drugs(35359,14367,144441,10592,and 28848)identified 33,6,12,37,and 34 cardiovascular AEs,and 16,5,7,25,and 21 instances of important medical events(IMEs)respectively.Each drug was linked to cases of cardiac and cardiopulmonary arrest.The cardiovascular AEs varied widely in occurrence and severity,with methadone notably presenting diverse and potent risks,including sudden cardiac death as a distinct medical event(DME).A comparison with SIDER 4.1 showed 11 opioid-related cardiovascular AEs in line with our findings.Standardized MedDRA Queries(SMQs)confirmed these results,indicating stronger signals for methadone and tramadol,while morphine,hydromorphone,and oxycodone exhibited fewer and weaker signals.Conclusion:The study revealed numerous heart-related adverse effects(AEs)not listed on drug labels and identified new AE patterns.Recognizing these differences in AE profiles and risks across different opioids is crucial for safer prescription practices to minimize cardiac complications.

Effectiveness of the“online+offline+practice”trinity teaching model on the learning of junior nursing students:an exploratory study

Background:Surgical Nursing is a main course of nursing specialty and a large course lasting 96 credit hours.In response to the teaching pain points such as the complicated and boring content of the surgical nursing course and the disconnection between classroom theory and clinical practice,surgical nursing is undergoing a teaching reform.Methods:The Surgical Nursing course builds trinity teaching model of“online+offline+practice”and implements teaching reforms by combining the Bridge,Objective,Pre-assessment,Participatory learning,Post-assessment,and Summary(BOPPPS)teaching concept.We constructed a trinity teaching model,and these teaching reform measures included building an online learning platform,offline case teaching and scenario simulation and hospital and community practice.The 50 students in the junior year of the undergraduate nursing program in the class of 2020 were used as the study subjects,and the students’comprehensive scores were compared.The questionnaire was also used to assess students’independent learning ability and to evaluate students’satisfaction with the teaching.Results:The students’course pass(≥60 points)rate was 100%,and the excellence(≥80 points)rate was 24.00%.Students’independent learning ability improved,and the scores and total scores of cognitive self-management ability,information ability and learning co-operation ability were significantly better than those of the national norm(P<0.01).The satisfaction of students in the BOPPPS group with classroom teaching was also significantly higher than that of the control group.Conclusion:The“online+offline+practice”trinity teaching mode can effectively integrate the“Rain Classroom”online platform with the BOPPPS teaching mode.It guides students to actively participate in classroom thinking and discussion,improves students’independent learning ability,and effectively enhances the effect of classroom teaching.

Hes1,an important gene for activation of hepatic stellate cells,is regulated by Notch1 and TGF-β/BMP signaling

AIM:To determine the role of Notch1 and Hes1 in regulating the activation of hepatic stellate cells(HSCs) and whether Hes1 is regulated by transforming growth factor(TGF)/bone morphogenetic protein(BMP) signaling.METHODS:Immunofluorescence staining was used to detect the expression of desmin,glial fibrillary acidic protein and the myofibroblastic marker α-smooth muscle actin(α-SMA) after freshly isolated,normal rat HSCs had been activated in culture for different numbers of days(0,1,3,7 and 10 d).The expression of α-SMA,collagen1α2(COL1α2),Notch receptors(Notch1-4),and the Notch target genes Hes1 and Hey1 were analyzed by reverse transcriptase-polymerase chain reaction.Luciferase reporter assays and Western blot were used to study the regulation of α-SMA,COL1α1,COL1α2 and Hes1 by NICD1,Hes1,CA-ALK3,and CA-ALK5 in HSC-T6 cells.Moreover,the effects of inhibiting Hes1 function in HSC-T6 cells using a Hes1 decoy were also investigated.RESULTS:The expression of Notch1 and Hes1 m RNAs was significantly down-regulated during the culture of freshly isolated HSCs.In HSC-T6 cells,Notch1 inhibited the promoter activities of α-SMA,COL1α1 and COL1α2.On the other hand,Hes1 enhanced the promoter activities of α-SMA and COL1α2,and this effect could be blocked by inhibiting Hes1 function with a Hes1 decoy.Furthermore,co-transfection of pc DNA3-CAALK3(BMP signaling activin receptor-like kinase 3) and pc DNA3.1-NICD1 further increased the expression of Hes1 compared with transfection of either vector alone in HSC-T6 cells,while pc DNA3-CA-ALK5(TGF-β signaling activin receptor-like kinase 5) reduced the effect of NICD1 on Hes1 expression.CONCLUSION:Selective interruption of Hes1 or maintenance of Hes1 at a reasonable level decreases the promoter activities of α-SMA and COL1α2,and these conditions may provide an anti-fibrotic strategy against hepatic fibrosis.

Dioscin-induced Apoptosis of Human LNCaP Prostate Carcinoma Cells through Activation of Caspase-3 and Modulation of Bcl-2 Protein Family

Dioscin is a natural steroid saponin derived from several plants, showing potent anti-cancer effect against a variety of tumor cell lines. In the present study, we investigated the anti-cancer activity of dioscin against human LNCaP cells, and evaluated the possible mechanism involved in its antineoplastic action. It was found that dioscin（1, 2 and 4 μmol/L） could significantly inhibit the viability of LNCaP cells in a time- and concentration-dependent manner. Flow cytometry revealed that the apoptosis rate was increased after treatment of LNCaP cells with dioscin for 24 h, indicating that apoptosis was an important mechanism by which dioscin inhibited cancer. Western blotting was employed to detect the expression of caspase-3, Bcl-2 and Bax in LNCaP cells. The expression of cleaved caspase-3 was significantly increased, and meanwhile procaspase-3 was markedly decreased. The expression of anti-apoptotic protein Bcl-2 was down-regulated, whereas the pro-apoptotic protein Bax was up-regulated. Moreover, the Bcl-2/Bax ratio was drastically decreased. These results suggested that dioscin possessed potential anti-tumor activity in human LNCaP cells through the apoptosis pathway, which might be associated with caspase-3 and Bcl-2 protein family.

Serum inter-cellular adhesion molecule 1 is an early marker of diagnosis and prediction of severe acute pancreatitis

AIM:To determine if serum inter-cellular adhesion molecule 1(ICAM-1)is an early marker of the diagnosis and prediction of severe acute pancreatitis(SAP) within 24 h of onset of pain,and to compare the sensitivity,specificity and prognostic value of this test with those of acute physiology and chronic health evaluation(APACHE)Ⅱscore and interleukin-6(IL-6). METHODS:Patients with acute pancreatitis(AP)were divided into two groups according to the Ranson's criteria:mild acute pancreatitis(MAP)group and SAP group.Serum ICAM-1,APACHEⅡand IL-6 levels were detected in all the patients.The sensitivity,specificity and prognostic value of the ICAM-1,APACHEⅡscore and IL-6 were evaluated. RESULTS:The ICAM-1 level in 36 patients with SAP within 24 h of onset of pain was increased and was significantly higher than that in the 50 patients with MAP and the 15 healthy volunteers(P<0.01).The ICAM-1 level(25 ng/mL)was chosen as the optimum cutoff to distinguish SAP from MAP,and the sensitivity,specificity,positive predictive value,negative predictive value(NPV),positive likelihood ratio and negative likelihood ratio were 61.11%,71.42%,0.6111,0.7142, 2.1382 and 0.5445,respectively.The area under the curve demonstrated that the prognostic accuracy of ICAM-1(0.712)was similar to the APACHE-Ⅱscoring system(0.770)and superior to IL-6(0.508)in distinguishing SAP from MAP. CONCLUSION:ICAM-1 test is a simple,rapid and reliable method in clinical practice.It is an early marker of diagnosis and prediction of SAP within the first 24 h after onset of pain or on admission.As it has a relatively low NPV and does not allow it to be a stand-alone test for the diagnosis of AP,other conventional diagnostic tests are required.

Saponins from Panax japonicus attenuate age-related neuroinflammation via regulation of the mitogen-activated protein kinase and nuclear factor kappa B signaling pathways

Neuroinflammation is recognized as an important pathogenic factor for aging and related cognitive disorders. Mitogen-activated protein kinase and nuclear factor kappa B signaling pathways may mediate neuroinflammation. Saponins from Panax japonicus are the most abundant and bioactive members in rhizomes of Panaxjaponicus, and show anti-inflammatory activity. However, it is not known whether saponin from Panaxjaponicus has an anti-inflammatory effect in the aging brain, and likewise its underlying mechanisms. Sprague-Dawley rats were divided into control groups （3-, 9-, 15-, and 24-month-old groups） and saponins from Panaxjaponicus-treated groups. Saponins from Panaxjaponicus-treated groups were orally administrated saponins from Panaxjaponicus at three doses of 10, 30, and 60 mg/kg once daily for 6 months until the rats were 24 months old. Immunohistochemical staining and western blot assay results demonstrated that many microglia were activated in 24-month-old rats compared with 3- and 9-month-old rats. Expression of interleukin-1β, tumor necrosis factor-a, cyclooxygenase-2, and inducible nitric oxide synthase increased. Each dose of saponins from Panaxjaponicus visibly suppressed microglial activation in the aging rat brain, and inhibited expression levels of the above factors. Each dose of saponins from Panax japonicus markedly diminished levels of nuclear factor kappa B, IKBa, extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38. These results confirm that saponins from Panaxjaponicus can mitigate neuroinflammation in the aging rat brain by inhibition of the mito- gen-activated protein kinase and nuclear factor kappa B signaling pathways.

Comparison of Treatment Outcomes of Ticagrelor and Clopidogrel among Patients Undergoing Percutaneous Coronary Intervention: A Meta-analysis

We performed a meta-analysis of randomized controlled trials（RCTs） to investigate the efficacy and safety of ticagrelor（TIC） vs. clopidogrel（CLO） in patients undergoing percutaneous coronary intervention（PCI）. In Jun 2016, a literature search was started and all the studies were conducted from 2010 to 2015. We systematically searched the literature through the MEDLINE database, Cochrane library, and EMBASE database. Quality assessments were evaluated with Jadad quality scale. Data were extracted considering the characteristics of efficacy and safety designs. Six RCTs enrolling 26 244 participants and satisfying the inclusion criteria were finally analyzed. There was a significant decrease of all-cause mortality（MD=0.83, 95%CI=0.74–0.93, P=0.001） and myocardial infarction（MI）（MD=0.78, 95%CI=0.70–0.88, P=0.000）. There were no significant differences in stroke（MD=1.34, 95%CI=0.99–1.79, P=0.06）, total bleeding（MD=0.97, 95%CI=0.84–1.12, P=0.66）, minor or major bleeding（MD=1.06, 95%CI=0.94–1.19, P=0.35） in patients undergoing PCI after treatment with TIC vs. CLO. TIC could be more significant in decreasing all-cause mortality and MI than CLO, but there were no significant differences between TIC and CLO in inhibiting stroke, major bleeding, major or minor bleeding in patients undergoing PCI.

Inhibitory Effects of Blockage of Intermediate Conductance Ca^(2+) -Activated K^+ Channels on Proliferation of Hepatocellular Carcinoma Cells

The roles of intermediate conductance Ca2＋-activated K＋ channel （IKCal） in the pathogene- sis of hepatocellular carcinoma （HCC） were investigated. Immunohistochemistry and Western blotting were used to detect the expression of IKCal protein in 50 HCC and 20 para-carcinoma tissue samples. Real-time PCR was used to detect the transcription level of IKCal mRNA in 13 HCC and 11 para-carcinoma tissue samples. The MTT assay was used to measure the function of IKCal in human HCC cell line HepG2 in vitro. TRAM-34, a specific blocker of IKCal, was used to intervene with the function of IKCal. As compared with para-carcinoma tissue, an over-expression of IKCal protein was detected in HCC tissue samples （P〈0.05）. The mRNA expression level of IKCal in HCC tissues was 2.17 times higher than that in para-carcinoma tissues. The proliferation of HepG2 cells was suppressed by TRAM-34 （0.5, 1.0, 2.0 and 4.0 pxnol/L） in vitro （P〈0.05）. Our results suggested that IKCal may play a role in the proliferation of human HCC, and IKCal blockers may represent a potential therapeutic strategy for HCC.

The Role of RP105 in Cardiovascular Disease through Regulating TLR4 and PI3K Signaling Pathways

Raidoprotective 105(RP105)was first discovered on the surface of mouse B cells and it has been demonstrated that RP105 can function as an inflammatory regulator in cardiovascular disease(CVD),such as myocardial ischemic reperfusion injury(MI/RI),atherosclerosis and myocardial infarction(MI).As a member of Toll-like receptor(TLR)homolog which is capable of regulating toll-like receptor(TLR4)signaling pathway,RP105 is implicated in various biological processes.Mounting evidence suggests that RP105 regulates the function of TLR4 and phosphoinositide 3-kinase(PI3K)signaling pathways.Here,we review the effect of RP105 on CVD through regulating TLR4/PI3K signaling pathways.

Effects of kanglaite capsules combined with transcatheter arterial chemoembolization （TACE） on patients with mid or late-stage primary hepatocellular carcinoma （HCC）

Objective： To observe the clinical effects of kanglaite （KLT） capsules combined with transcatheter arterial chemoembolization （TACE） in treating patients with mid or late-stage primary hepatocellular carcinoma （HCC）. Methods： Sixty-five cases were randomly divided into 2 groups, 32 patients in combination group received the treatment of KLT capsules ＋ TACE and 33 patients in control group were treated with TACE alone. The objective response rate （RR）, serum alpha fetoprotein （AFP）, peripheral blood T lymphocyte subgroups （T-LS）, quality of life （QOL）, time to progression （TTP） and adverse reaction were observed and compared between 2 groups. Results： The objective response rate and serum alpha fetoprotein levels had no significant difference between the two groups （P 〉 0.05）. Combination group was superior to control group in quality of life （QOL）, time to progression （TTP）, peripheral blood T lymphocyte subgroups （CD3＋, CD4＋, CD4＋／CD8 ratio） and liver adverse reactions, with significant differences （P 〈 0.05）. Conclusion： KLT capsules combined with TACE is an effective method to treat primary hepatocellular carcinoma （HCC） patients who have lost the opportunity of surgical therapy.

Roles of Human Epicardial Adipose Tissue in Coronary Artery Atherosclerosis

This study examined the adipocytokine-vascular interactions and link between epicardial adipose tissue and coronary artery atherosclerosis.Thirty-four patients undergoing open heart surgery were chosen randomly, and divided into groupⅠ(non-coronary artery disease group) and group Ⅱ(coronary artery disease group).Blood samples were taken through peripheral vein prior to surgery.Plasma levels of a panel of proteins (adiponectin, IL-10, TNF-α) were detected by using ELISA.Epicardial adipose tissue was taken near the proximal tract of the right coronary artery and subcutaneous adipose was taken from the leg before cardiopulmonary bypassing, adiponectin and CD68+ were detected by using RT-PCR and immunohistochemistry.Our results showed that plasma adiponectin level was significantly lower in the group Ⅱ as compared with group Ⅰ(P【0.05).There were no differences in plasma concentration (IL-10, TNF-α, tatal-chol, HDL-chol, LDL-chol) between group Ⅰ and group Ⅱ.The number of CD68+ cells in epicardial adipose tissue of group Ⅱ was significantly higher than that in subcutaneous adipose tissue.Adiponectin mRNA expression was 6 fold higher in subcutaneous adipose tissue than in epicardial adipose tissue of group Ⅱ (P【0.01).Furthermore, the level of adiponectin mRNA in the epicardial adipose tissue in group Ⅱ was also significantly lower than in group Ⅰ (P【0.05).We are led to conclude that inflammation that occurs locally in epicardial adipose tissue of CAD contributes to the pathogenesis of coronary artery disease.

Nogo-A and Nogo-A receptor expression in periventricular white matter of experimental autoimmune encephalomyelitis rats

BACKGROUND：Previous studies have focused on the correlation between Nogo-A expression and multiple sclerosis or between Nogo-A receptor （NgR） expression and multiple sclerosis in the central nervous system. Expression patterns of Nogo-A and NgR remain poorly understood in rat models of experimental autoimmune encephalomyelitis （EAE）.OBJECTIVE：To observe dynamic changes in Nogo-A and NgR protein expression, and to verify the correlation between Nogo-A and NgR protein, as well as expression patterns at various time points, in periventricular tissue of EAE rats.DESIGN, TIME AND SETrlNG：A neuroimmunological, randomized, controlled experiment was performed at the Clinical Institute of Hunan People＇s Hospital of China from September to November 2008.MATERIALS：Immunohistochemistry （streptavidin-biotin-peroxidase complex method） kit was purchased from Boster, China.METHODS：A total of 60 female, Wistar rats, aged 6-8 weeks, ware randomly assigned to EAE and control groups （n = 30, respectively）. Guinea pig spinal cord homogenate, self-made complete Freund＇s adjuvant （0.2 mL/100 g）, and pertussis vaccine （0.2 mL） were subcutaneously injected into the hindlimb foot pad of rats from the EAE group to create rat models of EAE. Complete Freund＇s adjuvant （0.2 mL） was infused into rats from the control group.MAIN OUTCOME MEASURES：Nogo-A and NgR protein expression was determined in periventricular white matter using immunohistochemical methods. Neurological scores ware determined in all rats.RESULTS：Rats from the EAE group developed acute-onset EAE following immunization. The pathogenetic symptoms reached a peak on day 15, and neurological scores ware also greatest at this time point. Neurological scores decreased with recovery of the illness. Nogo-A was shown to be expressed in neuronal cells and oligodendrocytes, and expression increased 11 days after immunization （P 〈 0.01）, decreased by day 13 （P 〈 0.01）, and then increased again by day 15. Nogo-A expression remained greater in the EAE group compared with the control group at day 30 （P 〈 0.01）. In the EAE group, NgR protein was primarily expressed on the surface of neuronal bodies and axons. NgR expression increased 13-18 days after immunization （P 〈 0.01 or P 〈 0.05）.CONCLUSION：Nogo-A and NgR protein expression altered with disease course in periventdcular white matter of EAE rats. Results suggested that Nogo-A and NgR were involved in EAE occurrence.

Renovascular Morphological Changes in a Rabbit Model of Hydronephrosis

Obstructive nephropathy ultimately leads to end-stage renal failure. Renovascular lesions are involved in various nephropathies, and most renal diseases have an ischemic component that underlies the resulting renal fibrosis. The aim of this study was to investigate whether morphological changes occur in the renal vasculature in hydronephrosis and the possible mechanisms involved. A model of complete unilateral ureteral obstruction（CUUO） was used. Experimental animals were divided into five groups： a normal control group（N） and groups of animals at 1st week（O1）, 2nd week（O2）, 4th week（O4） and 8th week（O8） after CUUO. Blood pressure was measured, renal arterial trees and glomeruli were assessed quantitatively, and renovascular three-dimensional reconstruction was performed on all groups. Glomerular ultrastructural changes were examined by transmission electron microscopy. The results showed that the systolic blood pressure was significantly increased in the obstructed groups（O1, O2, O4 and O8）. Three-dimensional reconstruction showed sparse arterial trees in the O8 group, and a tortuous and sometimes ruptured glomerular basement membrane was found in the O4 and O8 groups. Furthermore, epithelial media thickness and media/lumen ratio were increased, lumen diameters were decreased, and the cross-sectional area of the media was unaltered in the segmental renal artery, interlobar artery and afferent arterioles, respectively. In conclusion, renal arterial trees and glomeruli were dramatically altered following CUUO and the changes may be partially ascribed to vascular remodeling. Elucidation of the molecular mechanisms of renovascular morphological alterations will enable the development of potential therapeutic approaches for hydronephrosis.

Detection of Low-abundance Point Mutations by Competitive Strand Assisted Endonuclease Ⅳ Signal Amplification System

Genetic mutations are important molecular biomarkers for cancer diagnosis and surveillance. Therefore, the development of methods for mutation detection characterized with straightforward, highly specific and sensitive to low-level mutations within various sequence contexts is extremely needed. Although some of the currently available methods have shown very encouraging results, their discrimination efficiency is still very low. Herein, we demonstrate a fluorescent probe coupled with blocker and property of melting temperature discrimination, which is able to identify the presence of known or unknown single-base variations at abundances down to 0.1% within 20 min. The discrimination factors between the perfect-match target and single-base mismatched target are determined to be 10.15–38.48. The method is sequence independent, which assures a wide range of application. The new method would be an ideal choice for high-throughput in vitro diagnosis and precise clinical treatment.

Propofol application combined with ischemic preconditioning for aortic occlusion-induced spinal cord injury

BACKGROUND： Propofol combined with ischemic preconditioning （IPC） could prevent spinal ischemia/reperfusion injury. However, the effect of this combination remains poorly understood. OBJECTIVE： To measure neuroprotection of IPC in combination with propofol in a rabbit model of aorta occlusion-induced spinal injury. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at the Laboratory of Anesthesiology, Wuhan University and Central Laboratory of the First Clinical Medical College, China Three Gorges University, from October 2006 to April 2008. MATERIALS： Propofol was purchased from AstraZeneca, UK; malondialdehyde （MDA） and superoxide dismutase （SOD） kits were purchased from Nanjing Jiancheng Bioengineering Institute, China. METHODS： A total of 32 male, Japanese White rabbits, were randomly assigned to model, IPC, propofol, and combination groups, with eight rabbits in each group, using 2×2 factorial experimental design. Spinal ischemiaJreperfusion injury was induced by abdominal aorta occlusion for 40 minutes and reperfusion for 7 days. The IPC group was subjected to IPC treatment for 30 minutes prior to occlusion; the propofol group was treated with propofol for 10 minutes before occlusion; and the combination group underwent IPC treatment for 30 minutes and propofol for 10 minutes prior to occlusion. MAIN OUTCOME MEASURES： Serum MDA levels and SOD activity were detected 35 minutes prior to occlusion, immediately after reperfusion, and 60 minutes and 7 days after reperfusion, respectively. Rabbit hind limb nerve function and spinal pathological changes following injury were observed, and spinal neuronal apoptosis was determined using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method. RESULTS： Serum MDA levels, spinal neuronal apoptosis, and palsy incidence following injury were greatest in the propofol group, followed by the IPC and combination groups （P 〈 0.01）, while SOD activity and hind limb neurological scores were greatest in the combination group, followed by the IPC and propofol groups. Spinal cord injury in the combination group was slight （P 〈 0.01）. CONCLUSION： IPC and propofol treatment resulted in a synergistic effect for treating spinal ischemia/reperfusion injury. Combined application was superior to IPC or propofol treatment, suggesting that the protection of spinal cord injury may relate with anti-peroxidation.

Leukemia Inhibitory Factor Decreases Neurogenesis and Angiogenesis in a Rat Model of Intracerebral Hemorrhage

Neurogenesis and angiogenesis can improve the neurologic function after intracerebral hemorrhage(ICH).Leukemia inhibitory factor(LIF)plays an important role in neurogenesis and angiogenesis.In this study,a rat model of autologous blood-induced ICH was used to evaluate the effect of LIF on the neurogenesis and angiogenesis following ICH.After ICH,LIF-positive neurons and dilated vessels were detected in the peri-hematomal region.It was found that LIF levels increased significantly and peaked 14 days after ICH induction.Double immunofluorescence confirmed that LIF was expressed in neurons and endothelial cells.ICH also led to increases of doublecortin(DCX)-and von Willebrand factor(vWF)-positive cells as well as proliferation of cell nuclear antigen(PCNA)+/DCX+and PCNA+/vWF+nuclei.All these ICH-induced increases were significantly attenuated by exogenous LIF in fusion.These data suggested that LIF was a negative regulator of neurogenesis and angiogenesis after ICH.