Objective To explore the relationship of the expression level of monocyte Chemotactic protein-1 and nuclear factor κB(NF κB)with atrial fibrosis of atrial fibrillation patients with rheumatic heart disease.Methods About 200 mg right atrial tissue were taken from 26 patients with rheumatic heart disease undergoing valve replacement surgery and divided into sinus rhythm (SR) group (n=12) and atrial fibrillation (AF) group (n=14).Masson staining and atrial myocardial fibrosis markers were used to determine the level of fibrosis.The mRNA levels of cytokines in atrial tissue were measured by reverse transcription polymerase chain reaction (RT-PCR)The protein level of MCP-1 and phosphorylation of NFκB in atrial tissue were detected by Western blotting.Results Compared with SR group,the AF group showed that collagen volume fraction (AF:0.42 ± 0.03;SR:0.13 ± 0.02),the mRNA levels of myocardial fibrosis markers such as transforming growth factor (TGF)-β1,type Ⅰ collagen and type Ⅲ collagen,and cytokines such as IL-1β and TNF-α,and the protein level of MCP-1 (AF:0.170±0.003;SR:0.040±0.005) and level of phosphorylation of NF-κB (AF:0.35 ± 0.02;SR:0.12 ± 0.03) were significantly increased (all P＜0.05).In addition,the expression levels of cytokines,the protein expression level of MCP1 and the phosphorylation level of NF-κB were positively correlated with collagen volume fraction of the right atrial myocardial tissue (all P＜0.05).Conclusions Activation of NF-κB may induce the expression of MCP-1 in the myocardial tissue of patients with rheumatic heart disease,and sequentially stimulate the secretion of cytokines and extracellular matrix deposition,and finally participate in the occurrence and persistence of atrial fibrillation.

Objective To estimate the role of the posterior corneal parameters measured by Pentacam in screening subclinical keratoconus.Design Retrospective case-controlled study.Participants Forty five healthy subjects(73 eyes),14 keratoconus patients (20 eyes),43 subclinical keratoconus patients(58 eyes).Methods Based TOPOLYZER and biomicroscope signs,eyes were diagnosed as subclinical keratoconus and keratoconus to compare the differnces of related posterior corneal parameters provided by Pentacam in normal eyes and subclinical keratoconus or keratoconus eyes.Main Outcome Measures Back-Diff(6 mm) max,Back-Diff(6 mm )min, Back-Difference(6 mm),Back-Rmin,Back-Astig.Results The parameters which were statistically different between normal corneas and clinical keratoconus,normal corneas and subclinical keratoconus include Back-Diff(6 mm)max(P

Objective To investigate the scope of application of precise hepatectomy and its value.Methods The clinical data of 112 consecutive patients with liver neoplasm who received hepatectomy at Renji Hospital from November 2006 to March 2009 were retrospectively analyzed.Precise hepatectomy technique was applied to 88 patients(precise group),whereas pringle maneuver was applied to the rest 24 patients(prince group).Patients in precise group had undergone lobectomy,segmentectomy or local resection,while patients in pringle group received segmenteetomy or local resection.The perioperative conditions of patients in the 2 groups were compared via Fisher exact probability and l test.Results No perioperative mortality,hepatic failure,reoperation due to massive hemorrhage or bile leakage was observed.The blood transfusion rate,blood loss,postoperative total bilirubin(Tbil),prothrombin time(PT)were 7%(2/29),220 ml,20 μmol/L,13 seconds in patients who received segmentectomy and local resection in precise group,and were 4%(1/24),210 ml,19μmol/L and 13 seconds in patients who received segmentectomy and local resection in pringle group,with no significant difference(t=0.248,0.263,0.246,P>0.05).The operation time,postoperative white blood cell (WBC)count.alanine aminotransferase(ALT)value and fever incidence were 60 minutes,7.5×10~9/L,66 U/L,10%(3/29)in patients who received segmentectomy and local resection in precise group,and were 15 minutes,14.0×10~9/L,335 U/L and 42%(10/24)in patients who received segmentectomy and local resection in pringle group,with significant difference(t=4.962,4.961,4.959,P<0.05).In precise group,the blood transfusion rate,postoperative WBC count,ALT value.PT and fever incidence were 9%(4/45),8.3×10~9/L,153 U/L,17 seconds and 13%(6/45)in patients who received Iobectomy,and were 12%(5/43),8.2×10~9/L,133 U/L,14 seconds and 14%(6/43)in patients who received segmentectomy or lncal resection,with no significant difference (t=1.652,1.225,1.236,P>0.05);the blood loss,operation time and postoperative Tbil level were 350 ml,250 minutes and 32μmol/L in patients who received lobectomy.and were 240 ml,150 minutes and 21 μmol/L in patients who received segmentectomy or local resection(t=4.915,4.967,4.829,P＜0.05).Conclusions Precise hepatectomy can decrease damage to patients,but it should be applied in selected patients according to the tumor location and the excision range.Precise hepatectomy is recommended to be applied in lobeetomy.

With the application of monoclonal antibody technology more and more widely, its production technology is becoming more and more perfect. Small molecule monoclonal antibody technology is becoming a hot research topic for people. The application of traditional Chinese medicine small molecule monoclonal antibody technology has been more and more widely, the technology for effective Chinese medicine component knockout provide strong technical support. The preparation of monoclonal antibodies and small molecule knockout technology are reviewed in this paper. The preparation of several steps, such as: in the process of preparation of antigen, hapten carrier coupling, coupling ratio determination and identification of artificial antigen and establishment of animal immunization and hybridoma cell lines of monoclonal antibody, the large-scale preparation; small molecule monoclonal antibody on Immune in affinity chromatography column method is discussed in detail. The author believes that this technology will make the traditional Chinese medicine research on a higher level, and improve the level of internationalization of Chinese medicine research.
Animals ; Antibodies, Monoclonal ; chemistry ; genetics ; immunology ; Humans ; Hybridomas ; metabolism ; Immunologic Techniques ; methods ; trends

Animals ; Gene Expression Profiling ; Mitochondria, Heart ; genetics ; metabolism ; Myocardial Reperfusion Injury ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of graded hypothermia on neuropathologic alterations of neonatal rat brain after exposed to hypoxic-ischemic insult at 37°C, 33°C, 31°C, and 28°C, respectively, and to observe the effect of hypothermia on 72-kDa heat shock protein (HSP72) expression after hypoxic-ischemic insult.

METHODSSeven days old Wistar rats were subjected to unilateral common carotid artery ligation followed by exposure to hypoxia in 8% oxygen for 2 hours at 37°C, 33°C, 31°C, and 28°C, respectively. The brain temperature was monitored indirectly by inserting a mini-thermocouple probe into the temporal muscle during hypoxia. After hypoxia-ischemia their mortality was assessed. Neuronal damage was assessed with HE staining 72 hours after hypoxia. HSP72 expression at 0.5, 24, and 72 hours of recovery was immunohistochemically assessed using a monoclonal antibody to HSP72.

RESULTSHypoxia-ischemia caused 10.5% (2/19) of mortality in rat of 37°C group, but no death occurred in 33°C, 31°C or 28°C groups. HE staining showed neuropathologic damage was extensive in rats exposed to hypoxia-ischemia at 37°C (more than 80.0%). The incidence of severe brain damage was significantly decreased in 33°C (53.3%) and 31°C groups (44.4%), and no histologic injury was seen in the 28°C group of rats. Expression of HSP72 was manifest and persistent in the rat brain of 37°C group, but minimum in the rat brain of 28°C group.

CONCLUSIONMild and moderate hypothermia might prevent cerebral visible neuropathologic damage associated with hypoxic-ischemic injury by decreasing stress response.

Animals ; Animals, Newborn ; Body Temperature ; Female ; HSP72 Heat-Shock Proteins ; metabolism ; Hypothermia ; Hypoxia-Ischemia, Brain ; pathology ; Pregnancy ; Rats ; Rats, Wistar

Objectives: To evaluate mice as experimental animal for Chlamydia pneumoniae, a common cause of acute respiratory infections in human.　Methods: Intranasal inoculation of Icr mice with C. Pneumoniae induced a prolonged course of lung infection, as demonstrated by persistence of lung pathology(60 days).　Results: Icr mice were susceptible to C. pneumoniae. Lung pathology was characterized by patchy interstitial pneumonitis with predominately neutrophil leukocyte infiltration in the early(7 days) and lymphocytes infiltration in the later stages(14 days later) of infection.　Conclusions:Icr mice were susceptible to C. pneumoniae and the mouse model is useful for the investigation of the pathogenesis of C. pneumoniae infection.

Objective Analyzing the construction process and application of health appropriate technology database in Zhejiang province and proposing related considerations and recommendations for the database construction.Methods To design and construct the technology database system,and using excel software to calculate statistic data.Results Established a health appropriate technology database,which contains 826 technologies that can be selected and promoted by the demonstration bases in the whole province.Conclusions The establishment of the technology database increased the efficiency of health appropriate technology transformation at the provincial level,and also provided a reference for the national-wide generalizing of health appropriate technology.

OBJECTIVE In this study we explored the role of Epac1-Rap1 pathway in the acute myocardial ischemia/reperfusion injury (MIRI) in vitro and in vivo. METHODS An acute myocardial ischemia/reperfusion injury model was established by the ligation of left anterior descending coronary. Myocardial architecture, fibers and apoptosis was evaluated by the Masson trichrome staining, Sirius red staining and TUNEL assay.H9c2 cells were subjected to hypoxia for 5 h followed by 1-h reoxygen-ation in vitro.Cell viability was measured by MTT assay and cellular injury was evaluated by measuring the release of lactate dehydrogenase (LDH). Western blot, real-time PCR and immunofluorescence were used to detect the expressions of Epac1 and relative downstream molecules.RESULTS Myocardial IR-induced cardiac apoptosis and accumulation of Epac1 and Rap1 in rat IR injury model.Direct Epac activation by 8-CPT(8-(4-chlorophenylthio)-2′-O-methyl-cAMP)exacerbated cardiomyocyte death and dysfunction following hypoxia-reoxygenation(H/R),selective activation of Epac in response to H/R was evident which enriched for cytosolic/membrane proteins and mRNA. Harmacological inhibitor of Epac (ESI-09)significantly ameliorated myocardial injury with the decline of Epac expression.Epac inhibitor and agonist studies also implicated the effect of Rap1, which is downstream of Epac in this pathway. The expression of Rap1 elevated when activated by Epac agonist and was blocked by Epac inhibitor. The same result was true for myocyte CaMK-II and intracellular calcium ions activation.Moreover,ESI-09 also increased ERK1/2 phosphorylation. CONCLUSION Our study reveal that Epac1/Rap1 signaling pathway is involved in the pathogenesis of myocardial I/R injury in rats,which provides evidence on the development of therapeutic strategies target this pathway for myocardial I/R injury.

Background and purpose:LMP1 was one of the protein encoded by EBV latent gene,which was found to be able to transform cell lines and alter the phenotype of cells due to its oncogenic potential.In human epithelial cells,LMP1 alters many functional properties that are involved in tumor progression and invasions.In this study we investigated the influence of LMP1 silence on AP-1 signal transduction pathway and its downstream factors involved with cell transformation,proliferation and apoptosis.Methods:The chemically synthetic siRNA targeting LMP1 was transfected into EBV positive gastric carcinoma epithelial cell line by lipofectamine 2000 at 50 nmol/L final concentration.The protein expression of c-Jun,JunB and CDK4 was tested by,Western blotting.The mRNA of c-Jun,surviving,CDK4 and MMP9 mRNA were tested by RT-PCR.The expression of survivin were tested by immunohistochemistry.Results:Compared with the cell control,CDK4 mRNA was up regulated(P