2014年 · 第127卷第23期

Abstract：Background Our pervious antibacterial studies on several traditional Chinese medicines have found that Patchouli oil from Pogostemon cablin had significant antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA),which has spread worldwide and infected innumerable people.In order to find the more active natural substances in Patchouli oil,one of the major components,Pogostone,was isolated and its antibacterial activity was evaluated in vitro and in vivo in this study.Methods In vitro test,Pogostone was screened for antimicrobial properties against 83 bacteria comprising 35 gram positive and 48 gram negative bacteria strains via the agar double dilution method.In vivo test,specific pathogen free (SPF) strain of both male and female white Kunming mice,weighing 18-22 g,were used to test the protective ability of Pogostone after being injected with the median lethal doses (MLDs) of the tested strains.Results In vitro test,Pogostone could inhibit both gram negative bacteria (0.098-1 600 μg/ml) and gram positive bacteria(0.098-800 μg/ml).For Corynebacterium xerosis and some Chryseobacterium indologenes,the minimum inhibitory concentration (MIC) values of Pogostone were extremely low (＜0.098 μg/ml).It was significant that Pogostone was also active against some drug-resistant bacteria like MRSA.Furthermore,Pogostone showed antibacterial activity in vivo against Escherichia coli (E.coli) and MRSA via intraperitoneal injection.Ninety percent of the mice infected with E.coil could be protected at the concentrations of 50 and 100 mg/kg,and 60％ of the mice at 25 mg/kg,while the rate of protection for the mice infected with MRSA was 60％ and 50％ at doses of 100 and 50 mg/kg,respectively.Conclusion Pogostone could be developed as a potential antibacterial agent for clinical therapy。

Abstract：Background Hippophae rhamnoides L.(HL) exerts antioxidant activities against various oxidative stress conditions.In this study,we investigated effects of extract from HL leaves (HLE) on cell proliferation and neuroblast differentiation in the subgranular zone (SGZ) of the dentate gyrus (DG) of aged gerbils.Methods Aged gerbils (24 months) were divided into vehicle (saline)-treated-and HLE-treated-groups.The vehicle and HLE were orally administered with 200 mg/kg once a day for 20 days before sacrifice.Cell proliferation and neurobiast differentiation were examined in the DG using Ki67 and doublecortin (DCX),respectively.We also observed changes in immunoreactivities of superoxide dismutase 1 (SOD1) and superoxide dismutase 2 (SOD2),brain-derived neurotrophic factor (BDNF),and phospho-glycogen synthase kinase-3-beta (p-GSK-3β) to examine their relation with neurogenesis using immunohistochemistry.Results The administration of HLE significantly increased the number of Ki67-positive cells and DCX-positive neuroblasts with well-developed processes in the SGZ of the DG of the HLE-treated-group.In addition,immunoreactivities of SOD1,SOD2,BDNF,and p-GSK-3β were significantly increased in granule and polymorphic cells of the DG in the HLE-treated-group compared with those in the vehicle-treated-group.Conclusions HLE treatment significantly increased cell proliferation and neuroblast differentiation,showing that immunoreactivities of SOD1,SOD2,BDNF,and p-GSK-3β were significantly increased in the DG.These indicate that increased neuroblast differentiation neurogenesis may be closely related to upregulation of SOD1,SOD2,BDNF,and p-GSK-3β in aged gerbils。

Abstract：Background It has found that ischemic postconditioning (IPO) might decrease pulmonary ischemia/reperfusion (I/ R) injury,which is one of the main reasons of lung injury caused by cardiopulmonary bypass (CPB).It was found that aquaporins (AQPs) play a role in the maintenance of fluid homeostasis.But it is still unclear whether IPO influences the expression of aquaporin-1 (AQP1).This study was designed to investigate whether IPO can reduce CPB-related lung injury and affect the expression of AQP1 of lungs.Methods Twelve healthy dogs were divided into control group (C group) and ischemia postconditioning group (IPO group).CPB procedures were implemented.Ten minutes later,the left pulmonary artery was separated and blocked.Postconditioning consisted of two cycles of 5-minute pulmonary artery reperfusion/5-minute reocclusion starting at the beginning of reperfusion.The 2×4 cm tissues of both sides of pulmonary apex,superior,middle and inferior lobe were taken before CPB (T1),before occlusion and reopening of left pulmonary artery (T2,T3),and 2 hours after CPB (T4).Samples were used to evaluate lung injury degrees and to detect the expression of AQP1.At T1 and T4,blood was collected from femoral artery to calculate pulmonary function.Results At T4,each pulmonary function showed significant deterioration compared with T1.Lung injury could be found at the onset of CPB.However,the expression of AQP1 decreased and wet to dry weight ratio (W/D) increased after T2.In the left lung of C group,the worst pulmonary function and structures were detected.The slightest changes were discovered in the right lung of C group.A close relationship between W/D and lung injury score was found.The lung injury score was negatively related with the expression of AQP1.It was found that the expression of AQP1 was negatively connected with W/D.Conclusions In dog CPB models,lung injury induced by CPB was related with down regulated expression of AQP1.AQP1 is believed to be involved in the mechanisms of lung ischemia/reperfusion (I/R) injury caused by CPB.IPO increases the expression of AQP1,provides a protective effect on lung suffering from CPB,and alleviates CPB-related lung injury。

Abstract：Background It is still unclear whether the vitrification procedure itself is associated with the incidence of abnormal DNA methylation during oocytes vitrification.The purpose of this study was to evaluate the epigenetic profile of mouse oocytes,which went through vitrification either at a mature stage or at an immature stage following in vitro maturation (IVM) by analyzing the global DNA methylation.Methods Metaphase Ⅱ (M Ⅱ) stage and germinal vesicle (GV) stage oocytes were collected from adult female mice and were vitrified respectively.The M Ⅱ oocytes were assessed for cryo-survival and global DNA methylation.The GV oocytes were assessed for cryo-survival and only the surviving GV oocytes were cultured in vitro for subsequent assessment of global DNA methylation in mature oocytes.In vivo matured fresh M Ⅱ oocytes without undergoing vitrification were used as control.The level of global DNA methylation in the M Ⅱ oocytes was then examined by immunofluorescence using an anti-5-methylcytosine (anti-5-MeC) monoclonal antibody and fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG under a laser scanning confocal microscope.Results In terms of the effect of vitrification on global DNA methylation status in matured oocytes,in the M Ⅱ-v group,all the examined oocytes (90/90) were found with hypermethylation,including 63.3％ (57/90) of them displaying DNA methylation of a very high level,25.6％ (23/90) with a high level,and 11.1％ (10/90) with an intermediate level,whereas in the GV-v group,all the matured oocytes (129/129) were also examined with hypermethylation,including 67.4％ (87/129) of them displaying DNA methylation of a very high level,23.3％ (30/129) with a high level,and 9.3％ (12/129) with an intermediate level.Statistically,it was similar between both groups,which were similar to the control:68.6％ (83/121) of fresh M Ⅱ oocytes displayed DNA methylation of a very high level,21.5％ (26/121) with a high level,and 9.9％(12/121) with an intermediate level (P ＞0.05).In terms of the effect of IVM on global DNA methylation status in matured oocytes,in the in vivo matured oocytes group,all oocytes examined (94/94) were found with hypermethylation,including 80.9％ (76/94) displaying DNA methylation of a very high level and 19.1％ (18/94) with a high level,whereas in the in vitro matured oocytes group,all oocytes examined (69/69) were also found with hypermethylation:85.2％ (56/69) of them displayed with DNA methylation of very high level,11.9％ (11/69) with high level,and 2％ (2/69) with intermediate level.This result was similar to that in in vivo matured fresh M Ⅱ oocytes (P ＞0.05).Conclusion The vitrification procedure at GV stage does not induce widespread alteration of global DNA methylation status of mouse oocytes subsequently matured in vitro。

Abstract：Background Exposure of adult mice to more than 95％ O2 produces a lethal injury by 72 hours.Nuclear factor kappa B (NF-κB) is a transcriptional factor that plays a key role in the modulation of cytokine networks during hyperoxia-induced acute lung injury (ALl).Osteopontin (OPN) is a phosphorylated glycoprotein produced principally by macrophages.Studies have reported that exogenous OPN can maintain the integrity of the cerebral microvascular basement membrane and reduce brain damage through inhibiting NF-κB activities in the brain after subarachnoid hemorrhage.However,it is not clear whether OPN can reduce lung injury during ALl by inhibiting transcriptional signal pathways of NF-κB and consequent inhibition of infiammatory cytokines.Thus we examined the effects and mechanisms of recombinant OPN (r-OPN) on ALl.Methods Ninety-six mice were randomly divided into phosphate buffered saline (PBS) and r-OPN groups.Mice were put in an oxygen chamber (＞95％ O2) and assessed for lung injury at 24,48,and 72 hours.Expressions of NF-κB,matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9),and tissue inhibitors of MMP-2 and MMP-9 (TIMP-1,TIMP-2) mRNA in lungs were examined with RT-PCR.Expression and distribution of NF-κB protein in lungs were measured with immunohistochemistry.Results Exposure to hyperoxia for 72 hours induced more severe lung injury in the PBS group compared with the r-OPN group.Expression of NF-κB mRNA in the PBS group exposed to hyperoxia for 48 and 72 hours was significantly higher than the r-OPN group (P ＜0.05).With 72-hour exposure,expression of TIMP-1 mRNA in the r-OPN group was significantly higher than that of the PBS group (P ＜0.05).Expression of TIMP-2 mRNA in the r-OPN group at 48 and 72 hours was significantly higher than those in the PBS group (P ＜0.05).After 72-hour exposure,expression of NF-κB protein in airway epithelium in the PBS group was significantly higher than that in the r-OPN group (P ＜0.05).Conclusion r-OPN can inhibit the release and activation of MMPs through inhibition of the expression of NF-κB and promotion of the expression of TIMPs,and alleviate hyperoxia-induced ALl。

Abstract：Background The pathogenesis of benign prostatic hyperplasia (BPH) has been widely studied,and several biomarkers are known to play roles in its development.This study aimed to investigate the possible role of cysteine-rich protein 61 (CYR61),vascular endothelial growth factor (VEGF),androgen receptor (AR),interleukin-6 (IL-6),cytochrome c,caspase-3,and proliferating cell nuclear antigen (PCNA) in the clinical progression of BPH.Methods Tissue specimens from 96 BPH cases who underwent transurethral resection of the prostate were processed and transferred to tissue microarrays.Patient age,prostate volume,serum prostate-specific antigen (PSA) level,and International Prostate Symptom Score (IPSS) of all BPH cases were collected before surgery.The expression of CYR61,VEGF,AR,IL-6,cytochrome c,caspase-3,and PCNA was examined by immunostaining in the BPH specimens,and any possible correlation between the different biomarkers and risk factors for BPH clinical progression was analyzed.Results The expression of CYR61,VEGF,AR,IL-6,cytochrome c,caspase-3,and PCNA in the BPH cases was 68.8％ (66/96),77.1％ (74/96),43.8％ (42/96),31.3％ (30/96),35.4％ (34/96),56.3％ (54/96),and 29.2％ (28/96),respectively.The expression of both CYR61 and VEGF was positively correlated with patient age,prostate volume,and serum PSA level (P ＜0.05).Furthermore,cytochrome c and caspase-3 expression were inversely related to prostate volume (P ＜0.05),and AR expression was positively related to serum PSA level (P ＜0.05).Conclusion CYR61 and VEGF expression might serve as biomarkers for predicting the clinical progression of BPH due to effects on stromal cell proliferation and angiogenesis。

Abstract：Background Renal insufficiency (RI) is significantly associated with clinical prognosis in patients with heart failure (HF),but direct evidences on the relation between renal function and clinical outcome in patients receiving cardiac resynchronization therapy (CRT) are limited.The aim of the current study was to systematically evaluate the association of baseline and 6-month renal function with cardiac reverse remodeling and long-term outcome after CRT.Methods We retrospectively evaluated 190 consecutive patients who underwent CRT at Fuwai Hospital from January 2008 to April 2013.Renal function tests,echocardiographic measurement,and clinical parameters at baseline and after 6 months of CRT were performed.Primary endpoint events included all-cause mortality,cardiac transplantation,and unplanned hospitalizations for HF.Results At baseline,compared with normal renal function or mild RI (estimated glomerular filtration rate (eGFR) ＞-60 ml·min-1·1.73 m-2),moderate-to-severe RI (eGFR ＜60 ml·min-1·1.73 m-2) exerted a negative influence on cardiac reverse remodeling parameters.At 6-month follow-up,114 (60.0％) patients were classified as responders and showed significant renal function improvement,whereas renal function deteriorated in non-responders and subsequently 41 (25.6％) patients developed worsening renal function (WRF).During the mean follow-up of (24.3±17.1) months,both patients with baseline eGFR ＜60 ml.min-1·1.73 m-2 and those with WRF experienced worse event-free survival (P ＜0.01,respectively).Conclusions:This analysis identified that baseline eGFR as well as WRF after CRT were found to be independent determinants of the combined endpoints of all-cause mortality and HF-related hospitalizations in CRT recipients。

Abstract：Background Airway management is crucial in clinical anesthesia.Many complications associated with airway management result from unexpected difficult airway,but predicting a difficult airway is a major challenge.We investigated the efficacy of a new combined model including radiological indicators to predict difficult airway in patients undergoing surgery for cervical spondylosis,a population with a high incidence of difficult airway.Methods We randomly enrolled 303 patients scheduled for elective surgery for cervical spondylosis at Peking University Third Hospital between August 2012 and March 2013.Preoperatively,patients were evaluated for difficult airway according to a clinical index and parameters on lateral cervical radiographs and magnetic resonance images.Difficult airway was defined as Cormack-Lehane grades Ⅲ-Ⅳ.Logistic regression was used to identify a combined (clinical and radiological) model for difficult airway.A receiver operating characteristic (ROC) curve was used to describe the effectiveness of prediction.Results We identified three clinical predictive factors using the ROC curve:mouth opening,stemomental distance,and neck mobility.We created a clinical model using three factors:gender,age,and mouth opening,with odds ratios (OR) of 0.370,1.034,and 0.358,respectively.Using the clinical and radiological parameters,we formulated a combined model with five risk factors:gender,mouth opening,atlanto-occipital gap,the angle from the second to sixth cervical vertebraes in the neutral position,and the angle difference of δ (the angle between the laryngeal axis and the epiglottic axis) from the neutral position to extension (OR:0.107,0.355,0.846,1.057,and 0.952,respectively).The sensitivity and specificity of the combined model were 80.0％ and 65.7％,respectively,and the ROC curve confirmed that the combined model was better than any single clinical predictor and the clinical model.Conclusion The efficacy of the combined model including both clinical and radiological indicators was better than any single clinical predictor or the clinical model in patients undergoing elective surgery for cervical spondylosis。

Abstract：Background As a common form of gastric cancer migration,lymph node metastasis largely affects the surgical treatment and prognosis of gastric cancer.Surgery is the fundamental curative option for gastric cancer that varies depending on different stages.The study aimed to compare the clinicopathological characteristics and lymph node metastatic patterns in patients of proximal gastric cancer with different T stages and investigate a reasonable radical gastrectomy approach in terms of the range of lymphadenectomy for proximal gastric cancer.Methods In our retrospective study,the data of 328 patients of proximal gastric cancer with different T stages were analyzed.By comparing the differences of lymph node metastatic rate and ratio,we investigated the clinicopathological characteristics and metastatic patterns of lymph nodes.Also,we were especially interested in the differences in survival rates between patients with and without No.5 and 6 group metastasis with the same TNM stage.Results The overall lymph node metastatic rate and ratio of advanced proximal gastric cancer were 73.4％ and 23.3％,respectively.The tumors of different T stages were statistically significant in size and differentiation degree (P ＜0.05),multivariate analysis showed that the depth of tumor invasion was an independent risk factor for lymph node metastasis in proximal gastric cancer (RR,12.025; 95％ CI,2.326 to 62.157; P=0.003).The overall survival rate of patients with No.5,6 group lymph node metastasis and those without was significantly different,but the differences in survival rates between patients with and without No.5 and 6 group metastasis with the same TNM stage were not statistically significant.Conclusions Different T stages in proximal gastric cancer showed different patterns and characteristics of lymph node metastasis.D2 lymphadenectomy in patients with early gastric cancer had little survival benefit because metastasis to level 2 nodes was rare.Therefore the range of the lymph node dissection in radical gastrectomy for earty gastric cancer was considered reasonable.Moreover,to meet the requirements of the lymph node dissection,total gastrectomy plus D2 lymphadenectomy or more are supposed to be applied for the advanced proximal gastric cancer patients.Precise T staging larqely determines the range of gastrectomy and lymphadenectomy。

Abstract：Background Magnetic resonance imaging (MRI) has advantages in showing pathologic changes of disc space infection,which is important in clinical treatment.The purpose of this study was to describe the MRI findings of disc space infections in relation to chronicity.Methods MRI of 60 patients from January 1,2002 to April 30,2012 in Ottawa Hospital were retrospectively evaluated by two radiologists blindly.All patients had histological confirmation,with 55 having microbiological confirmation as well.These patients were divided into acute (n=18),subacute (n=21) and chronic (n=21) based on histological findings.The following potential signs of MRI finding were assessed:marrow edema,endplate erosions,disk fluid and height change,paraspinal mass,epidural collection,facet fluid and enhancement in the marrow,disc,paraspinal mass,and epidural involvement.Statistical analysis consisted of t-or F-tests and chi-square test.Results In the 60 patients,83 infected discs (single disc in 45 patients,2-4 discs in 15 patients) were found,including 22 discs in the acute group,30 discs in the subacute group,and 31 discs in the chronic group.There was a significant difference in the extent of marrow edema between the acute,subacute and chronic groups (P ＜0.05),with a gradually increasing extent from acute to chronic.The extent of endplate erosions increased with chronicity,but was not statistically significant.There were significant differences in the disc fluid,epidural collection,and disc enhancement among the acute,subacute and chronic groups,as well as the facet fluid between acute and chronic groups (P ＜0.05).There were no significant differences in the present probability of disc height loss,paraspinal mass,and marrow enhancement among the three groups (P ＞0.05).Conclusions From acute to chronic infections,the extent of marrow edema and endplate erosions appeared to gradually increase.Epidural collections and facet fluid are most frequently found in the acute group,while disc fluid and disc enhancement are more common in the chronic patients。

Abstract：Objective To give a concise review of the current state of the art in tissue engineering (TE) related to skeletal muscle and kinds of bioreactor environment.Data sources The review was based on data obtained from the published articles and guidelines.Study selection A total of 106 articles were selected from several hundred original articles or reviews.The content of selected articles is in accordance with our purpose and the authors are authorized scientists in the study of engineered muscle tissue in bioreactor.Results Skeletal muscle TE is a promising interdisciplinary field which aims at the reconstruction of skeletal muscle loss.Although numerous studies have indicated that engineering skeletal muscle tissue may be of great importance in medicine in the near future,this technique still represents a limited degree of success.Since tissue-engineered muscle constructs require an adequate connection to the vascular system for efficient transport of oxygen,carbon dioxide,nutrients and waste products.Moreover,functional and clinically applicable muscle constructs depend on adequate neuromuscular junctions with neural calls.Third,in order to engineer muscle tissue successfully,it may be beneficial to mimic the in vivo environment of muscle through association with adequate stimuli from bioreactors.Conclusion Vascular system and bioreactors are necessary for development and maintenance of engineered muscle in order to provide circulation within the construct。

Along with fast development of economy, public health demand and research budget, the medical research in China increased rapidly recent years, and the number of articles focusing on medicine from China, which were published by Journals indexed by Science Citation Index Expanded (SCI-E) or embodied in MEDLINE, increased obviously.~3 Chinese Medical Journal (CMJ) is a leading platform in China for following the progress of medical research in various disciplines from China and any part medical journal from the Chinese Medical Association, impact factor (IF) of CMJ increased to 1.016 in 2013 (Thomson Reuters, USA).

Interleukins and interferons are distinct groups of cytokines that are upregulated after viral infection in humans and have been recognized as having antitumor activity. For example, high-dose interleukin-2 (IL-2) has been used to treat malignant tumors, such as melanoma, in the clinic. In addition, interferon alfa-2b (IFNa-2b) has been applied extensively to treat hepatitis as well as renal cell carcinoma in the clinic. IFN~t-2b treatment can cause flu-like symptoms in patients receiving it and, in the worst cases, respiratory distress and acute interstitial pneumonia (AIP). AIP is a severe lung disease that typically progresses to respiratory failure。

Pneumocephalus is an uncommon, often forgotten complication of meningitis caused by gas-producing organisms, especially in advanced aged patients. Attention should be paid to physical signs, and early computed tomography (CT) scan could be helpful in the diagnosis. Accurate antibiotics according to blood culture would be the key point in the treatment procedure.

To the editor: We describe an unusual case of perforated metastatic malignant melanoma occurring ten years after diagnosis and treatment of cutaneous malignant melanoma in the patient's arm.