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

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 Dendritic cell-associatedC-type lectin-1 ( Dectin-1) is one of the most important receptors in antifungal innate immune response.This study was to construct a recombinant adenovirus vector expressing themurine Dectin-1gene and acquire a high-concentration adenovirus by amplification and purification. Methods The PCR amplification product CLEC7A-pIRES2-EGFP was cloned into the intermediate vector pDONR221, and then recom-bined with the backbone vector pAD/CMV/V5-DEST to produce a re-combinant plasmid pAD-CLEC7A-pIRE2S -EGFP.The recombinant plasmid was linearized with Pac I and transfected into human embryon-ic kidney ( HEK293) cells to produce recombinant adenovirus pAD-CLEC7Ap-IRES 2-EGFP. The adenovirus was propagated in the HEK293 cells and purified by filtering through the cellulose acetate membrane and concentrating column.Fluorescence microscopy and re-al-time PCR were used to determine the expression of the Dectin-1 gene. Results PCR identification, enzyme digestion, and sequen-cing results manifested theDectin-1 gene in the vector, with the final adenovirus titer of 5×1011 IU/mL.Fluorescence microscopy revealed green fluorescence and real-time PCR assay confirmed that the expression of Dectin-1 was improved by 8677.25 times. Conclusion A relatively high-titer adenovirus expressing Dectin-1 was acquired,which may help to further study the high expression of Dectin-1 in anti-fungal innate immunity in vitro and in vivo.

Objective Cryptococcosis is a potential life-threatening systemic mycosis with a heterogeneous susceptible popu-lation which is classified into three groups according to the current guidelines, including AIDS patients, organ transplantation recipients ( OTR) , and non-HIV-infected and non-transplant hosts ( NHNT) .This study aimed to discuss the influence of basic diseases on the clinical features and prognosis of NHNT cryptococcosis patients. Methods Using a retrospective cohort study design, we retrieved the clinical data about 73 NHNT cryptococcosis patients treated in Jinling Hospital.Based on the presence or absence of immunodefi-ciency or infection-increasing complications, we divided the patients into a basic disease group ( n=35) and a non-basic disease group ( n=38) and analyzed their clinical characteristics, chest radiographic features, laboratory results, and clinical outcomes. Results Compared with the non-basic disease group, the basic disease group showed a significantly higher incidence rate of disseminated disea-ses (62.9%vs 21.1%, P<0.01), more cases of patchy consolidation (47.4%vs 16.7%, P<0.05) and mixed lesion (31.6%vs 3.3%, P<0.05) in chest radiography, and a higher mortality (30.0%vs 5.3%, P=0.016). Conclusion Basic diseases have a great impact on the clinical features and prognosis of NHNT cryptococcosis.NHNT patients with basic diseases are susceptible to dis-seminated diseases with severer clinical symptoms and a higher mortality.

Objective To evaluate the effects of Ulinastatin (UTI) on the expressions of TNF-α,IL-6 and neurons apoptosis in cerebral cortex of rats after cardiopulmonary resuscitation (CPR).Methods Thirty-six healthy male adult Wistar rats were induced ventricular fibrillation untreated for 7 min and then received CPR.The animals were infused UTI 100 000 U/kg or phosphate-buffered solution (PBS) at once after ROSC.At 2,4 and 8 h after ROSC,cerebral cortex were removed to determine the mRNA expressions and levels of TNF-α protein and IL-6 protein,the translocation ratio of NF-κB p65 from cytoplasm to nucleus and the apoptotic neurons.Results The plasma levels of TNF-α (ng/mL) in animals of UTI group were (17.7 ± 1.4),(21.9 ± 2.1) and (17.1 ± 0.6) at 2,4 and 8 h after ROSC respectively,and significantly lower than those in PBS group at the given intervals.Mean while,the levels of IL-6 (ng/mL) were (208.9 ± 14.1),(281.5 ±25.9) and (251.8 ± 15.3) at 2,4 and 8 h after ROSC respectivèly in animals of UTI group,and lower than those in PBS group.The expressions of TNF-α mRNA and IL-6 mRNA and protein levels of TNF-α and IL-6 in UTI group were both lower than those in PBS group at given intervals,respectively.The translocation ratio of NF-κB p65 from plasma to nucleus in PBS group at each given interval after ROSC was significantly higher than that in UTI group.The number of viable neurons in cerebral cortex in UTI group was higher than that in PBS group,while the number apoptosis neurons was fewer in UTI group.Conclusions UTI attenuated the general inflammatory response after ROSC in rat,decreased the activation of NF-κB pathway,and subsequently attenuated the expression of TNF-α and IL-6,and finally decreased the neurons apoptosis.