Effect and Mechanism of Action of Epimedii Folium Polysaccharides on Mice with Exercise-induced Fatigue Based on p38 MAPK/NF-κB Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo study the effects of Epimedii Folium polysaccharides on mice with exercise-induced fatigue and explore its possible mechanism of action. MethodICR male mice screened by swimming training were randomly divided into a control group， model group， vitamin C group， and low， medium， and high dose groups of Epimedii Folium polysaccharides， with eight mice in each group. The exercise-induced fatigue model was established by weight-bearing swimming training in each group except for the control group. After two weeks of weight-bearing swimming， the Epimedii Folium polysaccharide groups were given 100， 200， 400 mg∙kg-1 of Epimedii Folium polysaccharides by gavage， and the vitamin C group was given 200 mg∙kg-1 of vitamin C by gavage. The control group and the model group were given equal amounts of saline for 14 d. At the end of the experimental period， the body mass of the mice in each group and the time of last swimming due to exhaustion were recorded. Serum urea nitrogen （BUN）， lactic acid （LA）， lactate dehydrogenase （LDH）， malondialdehyde （MDA）， superoxide dismutase （SOD）， glutathione peroxidation （GSH-Px）， myoglycogen （MG） in skeletal muscle， hepatic glycogen （HG） in the liver were detected by kits. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in muscle tissue. Western blot was used to detect the protein expression of p38 mitogen-activated protein kinase （p38 MAPK）， phosphorylation （p）-p38 MAPK， extracellular signal-regulated kinase1/2 （ERK1/2）， nuclear factor-κB （NF-κB）， p-NF-κB， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in muscle tissue. The immunofluorescence （IF） method was used to detect the expression of tumor necrosis factor-α （TNF-α） in skeletal muscle tissue of mice in each group. ResultCompared with the control group， the body mass of mice in the model group decreased， and the time of last swimming due to exhaustion decreased （P<0.01）. In addition， there were significantly higher serum levels of the fatigue metabolites LA， LDH， BUN， and lipid peroxidation product MDA （P<0.01） and decreased levels of MG， HG， SOD， and GSH-Px （P<0.01）. The protein expressions of p-p38 MAPK， ERK1/2， p-NF-κB， IL-1β， IL-6， and TNF-α in skeletal muscle tissue were significantly higher than those of the control group （P<0.01）. Compared with the model group， the body mass and time of last swimming due to exhaustion of the mice in the low， medium， and high dose groups of Epimedii Folium polysaccharides and the vitamin C group were increased （P<0.05， P<0.01）， and the contents of LA， LDH， BUN， and MDA were significantly decreased （P<0.05， P<0.01）. The levels of MG， HG， SOD， and GSH-Px increased （P<0.05， P<0.01）， and the protein expression levels of p-p38 MAPK， ERK， p-NF-κB， IL-1β， IL-6， and TNF-α in skeletal muscle tissue decreased （P<0.05， P<0.01）. ConclusionEpimedii Folium polysaccharides can play a role in alleviating exercise-induced fatigue by inhibiting the p38 MARK/NF-κB signaling pathway， thereby reducing the accumulation of metabolites， improving the activity of antioxidant enzymes， increasing the glycogen content of the body， and reducing inflammation in skeletal muscle.

Early whole body CT combined with coronary angiography for treatment guidance in patients with extracorporeal cardiopulmonary resuscitation / 中华急诊医学杂志

Objective:To investigate the safety of early whole body computed tomography (WBCT) combined with coronary angiography (CAG) in patients with extracorporeal cardiopulmonary resuscitation (ECPR) and its application value in the diagnosis of cardiac arrest and complications of cardiopulmonary resuscitation (CPR).Methods:This was a retrospective study. Patients who underwent ECPR in the Emergency Department of the First Affiliated Hospital of Nanjing Medical University from January 2017 to July 2021 were enrolled in this research. Patients younger than 18 years or with incomplete clinical data were excluded. The results of WBCT and CAG examinations after ECPR were collected.Results:A total of 89 patients with ECPR, aged (47±17) years, were enrolled in the study, all underwent WBCT examination, and no adverse events such as ECMO and tracheal tube shedding occurred. WBCT found 7 cases of pulmonary embolism, 3 cases of aortic dissection and 2 cases of cerebral hemorrhage. WBCT identified CPR-related complications in 42 cases, including rib fractures ( n=20), pneumothorax ( n=5), mediastinal emphysema ( n=5), subcutaneous emphysema ( n=6), and hematoma or swelling at puncture site ( n=6). Fifty-five patients underwent CAG examination, the most common culprit vessels were the left anterior descending branch disease (58.2%) followed by the left circumflex branch disease (27.3%), the right coronary artery disease (21.8%) and left main artery disease (12.7%). Conclusions:Early WBCT and CAG examinations are of great significance and safety for the guidance of treatment in ECPR patients.

Advances in pathogenesis and clinical features of HIV-associated neuroretinal disorder / 国际眼科杂志(Guoji Yanke Zazhi)

@#Ocular diseases in human immunodeficiency virus(HIV)infected patients have attracted increasing attention due to their impact on quality of life. As HIV treatment continues to improve, opportunistic eye infections are decreasing, while HIV-associated retinopathy is becoming a growing concern. HIV-associated retinopathy, including a series of structural changes in the retina and optic nerves(thinning of the nerve fiber layer, changing in blood vessels), has been found to cause decreased visual sensitivity, visual field defect, color vision disorder. However, the pathogenesis of HIV-associated neuroretinal disorder has not been fully clarified, and the existing findings may be related to direct destruction of retinal optic nerve tissue by HIV virus, chronic inflammation, and destruction of the blood-retina barrier. Understanding the pathological characteristics and possible mechanisms of HIV-associated neuroretinal disorder is expected to provide new ideas and approaches for the treatment of the disease and improve the quality of life of HIV-infected patients.

Predictive value of APACHEⅡ score combined with systematic inflammation markers on outcome in patients with extracorporeal membrane oxygenation / 中华急诊医学杂志

Objective:To investigate the clinical significance of the acute physiology and chronic health evaluationⅡ (APACHEⅡ) combined with different systematic inflammation markers (SIMs) including neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR)-in adult patients with venous-arterial extracorporeal membrane oxygenation (VA-ECMO).Methods:A total of 89 adult patients with VA-ECMO ( ≥ 3 d) in the Emergency Department of Jiangsu Provincial People's Hospital from January 2017 to June 2020 were retrospectively analyzed. Patients were divided into two groups: survivors ( n=39) and non-survivors ( n=50). The baseline APACHE Ⅱscore and PLR, NLR, LMR before ECMO implantation and at 1, 2, 3 day after ECMO were recorded. Binary logistic regression was used to analyze the risk factors of 28-day mortality in patients with VA-ECMO. The utility of APACHEⅡ score and SIMs alone or combination for predicting clinical prognosis was evaluated using receiver operating characteristic (ROC) curve analysis. The patients were divided into the high risk group and the low risk group according to the best cut-off value, and the difference of ECMO-related complications between the two groups was compared. Results:When combined APACHEⅡ score with SIMs, APACHEⅡ + PLR 48 h + LMR 24 h + LMR 72 h demonstrated the greatest predictive ability with an AUC of 0.833. Compared with the high-risk group, the low-risk group has a lower incidence of acute renal injury, infection, bleeding complications, the use of continuous renal replacement therapy, mechanical ventilation, and a higher hospital survival rate.Conclusions:The combination of APACHEⅡ score and SIMs-PLR, LMR- is better than a single one for death prediction, and it is expected to be a new predictive model for early identification of the risk of death or poor prognosis in patients with VA-ECMO.

Predictive value of SAVE score combined with 24-h lactate on the prognosis of patients with extracorporeal cardiopulmonary resuscitation / 中华急诊医学杂志

Objective:To explore the prognostic value of survival after veno-arterial ECMO (SAVE) score combined with 24-h lactate on the machine in patients with extracorporeal cardiopulmonary resuscitation (ECPR).Methods:Totally 59 patients treated with ECPR in the Emergency Department of the First Affiliated Hospital of Nanjing Medical University from April 2017 to June 2021 were retrospectively analyzed. According to the 28-day prognosis, the patients were divided into the death group ( n=36) and the survival group ( n=23). The differences in baseline data were analyzed, and multivariate logistic regression was performed to identify the influencing factors of 28-day mortality in patients with ECPR. The receiver operating characteristic (ROC) curve was applied to evaluate the predictive value of SAVE score, 24-h lactate and their combined detection for predicting 28-day mortality risk in patients with ECPR. Results:The 28-day survival rate of patients with ECPR was 39% (23/59). SAVE score of the death group was significantly lower than that of the survival group (-11.67±4.60 vs. -2.43±4.77, P<0.001), and the 24-h lactate in the death group was significantly higher than that in the survival group [5.94 (3.37, 12.40) mmol/L vs. 1.65 (1.07, 3.15) mmol/L, P<0.001]. Multivariate logistic regression analysis showed that SAVE score ( OR=0.703, 95% CI: 0.566-0.873, P=0.001) and 24-h lactate ( OR=1.608, 95% CI: 1.025-2.523, P=0.039) were independent influencing factors of 28-day mortality in ECPR patients. ROC curve analysis showed that the best cut-off value of SAVE score was -6, the sensitivity was 78.30% and specificity was 91.70%. The best cutoff value of 24-h lactate was 4.7 mmol/L, the sensitivity was 63.90% and specificity was 100.00%. The sensitivity and specificity of the combined detection of SAVE score and 24-h lactate were 82.60% and 100.00%, respectively. The area under the curve (AUC) of SAVE score combined with 24-h lactate for predicting the 28-day mortality risk in patients with ECPR was larger than that of SAVE score and 24-h lactate alone (0.952 vs. 0.917; 0.952 vs. 0.847). Conclusions:Lower SAVE score and higher 24-h lactate are independently risk factors of 28-day mortality in patients with ECPR, and SAVE score combined with 24-h lactate on the machine has a good predictive value for the prognosis of patients with ECPR.

Analysis of the development trend of small nucleic acid drugs / 中国药房

OBJECTIVE To know about the development trend of small nucleic acid drugs in the world ，to provide reference for the research and development of small nucleic acid drug in China. METHODS By searching the academic literature and patents related to small nucleic acid drugs through the Web of Science literature database and PatSnap patent database from Jan. 1980 to Dec. 2021，research and development situation of small nucleic acid drugs were revealed comprehensively by analyzing research enthusiasm，R&D countries ，R&D institutions and technical topics of small nucleic acid drugs. RESULTS & CONCLUSIONS A total of 59 819 documents and 37 645 patent groups were included. The global trend of small nucleic acid drug literature publication and patent application could be divided into three stages. From 2003 to 2021，the research enthusiasm for small nucleic acid drugs continued to increase. The United States ，China，Japan and Germany were the main research and development countries for small nucleic acid drugs. The number of document publications （25 703，15 927 papers）and patent applications （14 240、8 937 groups） in the United States and China were ahead of other countries ，and the research and development activities were relatively strong. Moreover，the number of document publications and patent applications in China in this field had grown rapidly in recent years. The R&D institution with the largest number of publications was the University of California （2 499 papers），the R&D institution with the largest number of patent applications was the American Ionis Corporation （1 378 groups），and the Chinese Academy of Sciences （1 580 papers）had been shortlisted among the top 10 document producing institutions in the world. However ，our country ’s research and development in this field are mostly based on basic research ，and the research on industrial application is slightly insufficient. The research focus in the field of small nucleic acid drugs mainly focuses on nucleic acid sequences and their modification and improvement and drug loading technology. RNA interference technology has gradually become a hot technology for small nucleic acid drugs.

The prognostic value of platelet-to-lymphocyte ratio in patients with extracorporeal membrane oxygenation / 中华急诊医学杂志

Objective:To analyze the potential role and prognostic value of platelet-to-lymphocyte ratio (PLR) at an early stage in arterial-venous extracorporeal membrane oxygenation (VA -ECMO).Methods:Totally 83 adult patients with VA-ECMO from June 2018 to June 2020 treated at Emergency Department of Jiangsu Provincial Hospital were retrospectively analyzed. Baseline characteristics between survivors ( n=46) and non-survivors ( n=37) were compared. Logistic regression analysis was used to predict the risk factors associated with 28-day mortality in VA-ECMO patients. The cut-off value was calculated by the receiver operating characteristic (ROC) curve. Results:PLR48-h ( OR=1.018,95% CI: 1.001-1.036, P=0.039) and continuous renal replacement therapy (CRRT) ( OR=7.095,95% CI: 1.099-45.799, P=0.039) were relevant risk factors of 28-day mortality in VA-ECMO patients. The cut-off value of PLR48-h was 156.3 [sensitivity: 57.8%, specificity: 86.1%, and area under the curve (AUC): 0.756]. Compared with the high PLR group (>156.3), the incidences of acute kidney injury (AKI) ( P<0.001) and bleeding events ( P=0.013) were significantly higher in the low PLR group (<156.3). Conclusions:The early PLR reduction and CRRT application during VA-ECMO support are related to poor prognosis.

Enzymatic Hydrolysis Characteristics of Five Medicinal Starches by Gravitational Field-Flow Fractionation / 中国药学杂志

OBJECTIVE: To investigate the enzymatic hydrolysis characteristics of five medicinal starches, i.e.,waxy corn starch, potato starch, pea starch, wheat starch and common corn starch,using gravitational field flow fractionation (GrFFF) and other technologies. METHODS: Firstly, the enzymatic hydrolysis rates of the five medicinal starches were characterized by GrFFF, and the results of GrFFF were verified by determination of the reducing sugar.Secondly, scanning electron microscopy (SEM) and particle size analyzer were used to characterize the changes of particle morphology and size distribution during amylase hydrolysis. RESULTS: The experiments showed that the enzymatic hydrolysis rates of the five medicinal starches were successively decreased. The former two as well as the latter three reached the maximum enzymatic rate at 6-12 h and 12-24 h, respectively. As the enzymatic hydrolysis time increased, the particle size distribution range became wider and the average size increased. The results of reducing sugar quantitation, SEM and particle size analysis were almost identical to those of GrFFF. CONCLUSION: This study proves that GrFFF is an effective analytical technology to characterize the enzymatic hydrolysis of medicinal starch, which expands the application range of GrFFF and provides useful reference for the application of starch in medicine and food sciences.

Dosimetric comparison between three radiotherapy regimens involving supraclavicular and infraclavicular regions in breast cancer patients after breast-conserving surgery / 中华放射肿瘤学杂志

Objective To evaluate the dosimetric difference between fixed-field static intensity-modulated radiotherapy (IMRT), fixed-field dynamic multileaf collimator (DMLC), and volumetric modulated arc therapy (VMAT), all of which involve supraclavicular and infraclavicular regions, in breast cancer patients after breast-conserving surgery.Methods This study included 14 female patients with breast cancer who received radiotherapy after breast-conserving surgery in our hospital from October 2012 to April 2016.The radiation field included the chest wall and supraclavicular and infraclavicular regions.IMRT, DMLC, and VMAT plans were generated for each patient while using identical optimization conditions.The doses to planning target volume (PTV) and organs at risk (OARs) were compared based on dose-volume histogram (DVH);one-way analysis of variance or nonparametric Wilcoxon rank test was used for comparison.Results For the dose distribution of PTV, VMAT achieved the best V95, V98, CI, and HI (P<0.009).Concerning the doses to OARs, VMAT achieved the best V5, V20, and Dmean of the ipsilateral lung and the best V5 and Dmean of the contralateral lung (P<0.022).Dmean of the spinal cord was significantly lower in VMAT than in IMRT and DMLC (P=0.004).Conclusions VMAT is preferred for the patients with breast cancer to be treated with radiotherapy involving supraclavicular and infraclavicular regions after breast-conserving surgery.It can improve the dose distribution of target and reduce the doses to organs at risk and radiotherapy toxicities.

Effect of tumor volume and radiation dose on the prognosis of hepatocellular carcinoma patients treated with whole body gamma knife / 中华放射肿瘤学杂志

Objective To examine the effects of gross tumor volume (GTV) and radiation dose on the prognosis of hepatocellular carcinoma (HCC) patients treated with whole body gamma knife.Methods The clinical data of 69 HCC patients who underwent body gamma knife treatment from January 2012 to June 2015 in the Radiotherapy Center of the PLA General Hospital were retrospectively reviewed.Based on a 50% or 60% isodose coverage of the planning target volume (PTV), patients were treated with a radiation dose of 4-5 Gy per fraction, and a total marginal dose of 36-50 Gy (median dose 45 Gy).Short-term efficacy, overall survival (OS), and the adverse effect of the treatment were evaluated.The optimal cut-off tumor volume was identified using the receiver operating characteristic curve, and survival was determined by the Kaplan-Meier method.Univariate and multivariate analyses were performed using the log-rank test and Cox proportional hazards regression model, respectively.Results The overall short-term response rate of the 69 patients was 67%.The 1-and 2-year OS rates were 62% and 40%, respectively, with a median survival of 18.6 months.The multivariate analysis showed that gross tumor volume (GTV)93 cm3(P=0.665).Conclusions GTV is an independent prognostic factor for overall survival of HCC patients.Although high-dose radiotherapy provides survival benefits to patients with small GTV, it is not necessarily suitable for patients with large GTV.

Administration of aspirin and rivaroxaban prevents deep vein thrombosis after total knee arthroplasty / 中国组织工程研究

BACKGROUND:To date, rivaroxaban has been a clinical y common anticoagulant in China;however, effective prophylaxis for venous thrombosis is associated with a markedly higher incidence of perioperative hemorrhagic complications. Although it has been reported that aspirin effectively prevents deep vein thrombosis and pulmonary embolism, the use of aspirin as a routine drug for venous thrombosis after total knee arthroplasty is stil controversial. OBJECTIVE:To compare the efficacy and safety of aspirin and rivaroxaban for prevention of deep vein thrombosis after total knee arthroplasty. METHODS:Total y 324 patients with osteoarthritis who underwent primary unilateral total knee arthroplasty were randomly divided into three groups. Twelve hours after the surgery, three groups were given aspirin, rivaroxaban and low-molecular-weight heparin respectively. Al three groups were treated for 14 days, and al of the patients were fol owed for 4 weeks. RESULTS AND CONCLUSION:Compared with the low-molecular-weight heparin group, the incidence of deep vein thrombosis was lower (P0.05). The results confirmed that rivaroxaban has a positive anticoagulation effect but leads to increases in wound complications in patients;there are no differences in efficacy and safety between aspirin and low-molecular-weight heparin, so aspirin as part of a multimodal anticoagulation therapy after total knee arthroplasty has good clinical safety and efficacy.

PI3K/Akt-mediated promotive effect of insulin on proliferation of retinal pigment epithelial cells and secretion of transforming growth factor-β2 / 中华实验眼科杂志

Background Insulin can promote the occurrence of myopia.It has been proven that insulin receptor exists in human retinal pigment epithelial (RPE) cells and can promote RPE cells to secrete transforming growth factor-β2(TGF-β2),which is one of the most important myopic signal molecules.Objective This study was to investigate if PI3K/Akt mediates the promotive effects of insulin on proliferation of human RPE cells and secretion of TGF-β2.Methods Human RPE cell line,ARPE-19 cells,were regularly cultured using DMEM containing 10％ fetal bovine serum,and 10× 103 U/ml insulin,LY294002,10× 103 U/ml insulin+LY294002,Wortmanin,10× 103 U/ml insulin+Wortmanin were added into the medium respectively for 48 hours,and the regularly cultured cells served as blank controls.The proliferation value (absorbance,A) of the cells was evaluated by MTS,and the TGF-β2 level in the cell supernatant was detected by ELISA.The relative expression of TGF-β2 mRNA in the cells was assayed using reverse transcription PCR (RT-PCT) 1 hour and 2 hours after the addition of reagents.Results MTS showed that the proliferation value of the cells in the insulin+LY294002 group was 0.75±0.03,which was significantly lower than that in the insulin group (0.98± 0.04).No significant difference was seen in the proliferative value between the insulin+Wortmanin group and the insulin group (0.97±0.07 versus 0.98± 0.04,P＞0.05).ELISA revealed that the content of TGF-β2 in the the cell supernatant was (11.59±2.85) pg/ml and (49.16± 10.94) pg/ml in the insulin + LY294002 group and the insulin + Wortmanin group,respectively,showing a significant decline in comparison with (548.50±35.18) pg/ml in the insulin group (both at P＜0.05).A significant difference was found in the TGF-β2 content between the insulin+LY294002 group and the insulin+Wortmanin group (t =8.131,P =0.000).The RT-PCR showed that 1 hour and 2 hours after addition of the reagents,the expression levels of TGF-β2 mRNA in the cells were lower in both insulin+LY294002 group and insulin+Wortmanin group than those in the insulin group (P＜0.05).The decline range of TGF-β2 mRNA expression level was more significant in the insulin+LY294002 group than that in the insulin+Wortmanin group at 1 hour (t=4.176,P=0.014) rather than at 2 hours (t=0.756,P=0.492).Conclusions Insulin can promote the proliferation of human RPE cells and secretion of TGF-β2 through PI3K/Akt pathway.This may be one of the mechanisms of insulin causes myopia.

A dosimetric comparison of three-dimensional conformal and intensity-modulated radiotherapy in locally advanced pancreatic cancer and a study of the clinical efficacy / 中华放射医学与防护杂志

Objective To compare the dose distribution between three-dimensional conformal radiotherapy(3D-CRT) and intensity-modulated radiotherapy (IMRT) in treating locally advanced pancreatic cancer,and report the efficacy of IMRT combined with regional chemotherapy using gemcitabine (GEM).Methods Ten patients with locally advanced pancreatic cancer were enrolled in this study.3D-CRT and IMRT plans were designed for each patient.The dose distributions of target volume and normal tissues were analyzed using the dose volume histogram (DVH).Twenty-five locally advanced pancreatic cancers patients who were treated by IMRT combined with regional chemotherapy using gemcitabine (combined group) were retrospective analyzed,as well as 25 hospitalized patients of the same period who were treated by regional chemotherapy using gemcitabine alone (chemotherapy alone group).The therapeutic efficacy and adverse events were compared between two groups.Results IMRT plans decrease the mean dose and volume of duodenum,liver,stomach,both kidney and small bowel that received highdose irradiation.The 1-,2-year survival rate of the combined group and chemotherapy alone group was 60％,28％ and 36％,12％.The median survival time of two groups was 15 and 10 months,respectively (x2 =4.16,P ＜0.05).The total response rate of the combined group and the chemotherapy alone group was 64％ and 32％,respectively (x2 =5.13,P ＜ 0.05).The upper gastrointestinal side-effect rate of the combined group was higher than that of the chemotherapy alone group(Z =-2.354,P ＜ 0.05).There was no statistic significance in hematologic toxicity,liver and renal functional damage between the two groups.Conclusions Compared with 3D-CRT plan,IMRT plan could reduce the dose of organ at risks.IMRT combined with regional chemotherapy using gemcitabine could significantly improve the survival rate of patients with locally advanced pancreatic cancer with mild adverse events.

Effects of X-ray irradiation on the expression of Pokemon gene in human lung adenocarcinoma cell line / 中华放射肿瘤学杂志

ObjectiveTo study the dose and time effects of X-ray radiation on the expression of Pokemon gene and protein in human lung adenocarcinoma cell line A549.MethodsA549 cells was exposed to different doses of X-ray (2,4,6 and 8 Gy),and the expression of Pokemon mRNA and protein of the cells was detected by using Quantitative real-time PCR and western-blotting at 2,4,8,12,24,and 48 h after irradiation. 3-( 4,5-Dimethylthiazol-2-yl )-2,5-diphenyltetrazoliumbromidewasused to detectthe proliferation of A549 cells at 1,2,3,4,and 5 d after 2 Gy X-ray irradiation.The mock treated A549 cells were used as the control.ResultsThe expression of Pokemon mRNA trended to decrease after irradiated with 4,6 and 8 Gy in the earlier period and increased in the later period with statistical difference at the most time points (t =3.40 -154.76,P =0.000 -0.041 ).The expression of Pokemon protein trended to increase and reached the peak at 8 h after irradiated of 2,4,6 and 8 Gy with statistical difference at the most time points ( t =4.18 - 89.64,P =0.000 - 0.039).Compared with the control,the proliferation of A549 cells was significantly inhibited during 3 to 5 d after irradiation of 2 Gy ( t =2.34 - 18.19,P =0.000 -0.040).ConclusionsX-ray irradiation may increase the expression of Pokemon mRNA and protein in A549 cells,which might be correlated with radiation-resistance of A549 cells.

Earlier detection of tumor treatment response using magnetic resonance diffusion imaging with oscillating gradients.

An improved method for detecting early changes in tumors in response to treatment, based on a modification of diffusion-weighted magnetic resonance imaging, has been demonstrated in an animal model. Early detection of therapeutic response in tumors is important both clinically and in pre-clinical assessments of novel treatments. Noninvasive imaging methods that can detect and assess tumor response early in the course of treatment, and before frank changes in tumor morphology are evident, are of considerable interest as potential biomarkers of treatment efficacy. Diffusion-weighted magnetic resonance imaging is sensitive to changes in water diffusion rates in tissues that result from structural variations in the local cellular environment, but conventional methods mainly reflect changes in tissue cellularity and do not convey information specific to microstructural variations at sub-cellular scales. We implemented a modified imaging technique using oscillating gradients of the magnetic field for evaluating water diffusion rates over very short spatial scales that are more specific for detecting changes in intracellular structure that may precede changes in cellularity. Results from a study of orthotopic 9L gliomas in rat brains indicate that this method can detect changes as early as 24 h following treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea, when conventional approaches do not find significant effects. These studies suggest that diffusion imaging using oscillating gradients may be used to obtain an earlier indication of treatment efficacy than previous magnetic resonance imaging methods.

Effects of X-rays on CC-chemokine receptor 7 expression in human lung cancer A549 cells / 中华放射医学与防护杂志

Objective To study the effects of X-ray radiation on CC-chemokine receptor 7(CCR7) expression in human non-small cell lung cancer (NSCLC) cells.Methods Humanadenocarcinoma cells of the line A549 were cultured and irradiated by X-ray at the absorbed doses of 2,4,6,and 8 Gy respectively by linear accelerator (with the source skin distance of 100 cm and dose rate of 442.89 cGy/min).The relative levels of CCR7 mRNA and protein expression in the A549 cells were respectively detected by real time-PCR and Western blotting 4,12,24,48,and 72 h after radiation.Untreated A549 cells were used as control group.Results The expression levels of CCR7 mRNA and protein in the A549 cells began to increase since 4 h after radiation and then decreased gradually after they reached the peak.The CCR7 mRNA expression levels 72 h after radiation of the 6 and 8 Gy groups were still significantly higher than those of the control group (t = 6.75-7.26,both P < 0.01),and the CCR7 protein expression levels of the 2 and 6 Gy group were still significantly higher than those of the control group(t=11.13-14.17,both P <0.01).Then the CCR7 protein expression levels of the 4 and 8 Gy groups decreased to the control group level 48 and 72 h after radiation respectively.Conclusions The CCR7 mRNA and protein expression levels in the NSCLC cells increase after X-ray irradiation,which may be correlated with the promotion of proliferation and metastasis of NSCLC cells by X-ray irradiation at a certain dose.

Effects of X-ray irradiation on expression of Pokemon gene in A549 cells of human lung adenocarcinoma / 中华放射医学与防护杂志

Objective To study the dose-and time-effects of X-ray irradiation on the expression of Pokemon gene in A549 cells of human lung adenocarcinoma.Methods A549 cells were cultured in vitro and exposed to X-rays with the doses of 2,4,6 and 8 Gy,respectively.Untreated A549 cells were used as control group.The relative levels of Pokemon mRNA expression in the cells were detected by using quantitative real-time PCR at 2,4,8,12,24,48 and 72 h after irradiation.Results The Pokemon mRNA expression levels decreased in the early period after irradiation(except 2 and 4 h after irradiation in 2 Gy group)and then increased in the later stage(48 h after irradiation)with significant statistical differences at the most time points in comparison with the control group(t=3.40-154.76,P<0.05).Conclusions Higher doses of X-rays may degrade the expression of Pokemon mRNA in the human A549 cells and induce apoptosis in the early period,hut also may upgrade its expression in the later period, which might be correlated with the cell cycle regulation and DNA damage repair in the A549 cells.

Rat blood collection methods / 中国组织工程研究

BACKGROUND: A total of 300 SD rats were used for blood sampling. In abdominal aorta approach, the best puncture point was the abdominal aortic bifurcation 1-3 mm towards the heart, with a success rate of 93.6%. In posterior orbital venous plexus approach, the needle was vertically inserted into the inner canthus and rotated toward the eyeground to open venous plexus (success rate 89.9%). In cardiac puncture approach, below the xiphoid process, the needle punctured into the skin with 25°-30° oblique upward, through the diaphragm until 2.5-3.0 cm deep (success rate 83.4%). In tail end approach, surgical scissors cut off 5-10 mm tail top (success rate 94.4%). In jugular vein approach, the needle was horizontally inserted along the fourth rib into the skin until the jugular vein, about 5 mm deep, at 30°-40° with the chest surface (success rate 80.9%). A large blood volume could be obtained by abdominal aorta approach, which leads to less haemolysis and no hurt to organs, no gas embolism or haemostasis caused by inappropriate operation. But each approach has advantages and drawbacks, the selective principle should be based on experimental require.

A zebrafish model of human Barth syndrome reveals the essential role of tafazzin in cardiac development and function.

Barth syndrome is an X-linked disorder characterized by cardiomyopathy, skeletal myopathy, neutropenia, organic aciduria, and growth retardation caused by mutations in tafazzin. The sequence similarity of tafazzin to acyltransferases suggests a role in mitochondrial phospholipid metabolism. To study the role of tafazzin in heart function and development, we created a knockdown zebrafish model. Zebrafish tafazzin mRNA is first evident at 7 hours post-fertilization (hpf). At 10 and 24 hpf, tafazzin mRNA is ubiquitous, with highest levels in the head. By 51 hpf, expression becomes cardiac restricted. The tafazzin knockdown created by antisense morpholino yolk injection resulted in dose-dependent lethality, severe developmental and growth retardation, marked bradycardia and pericardial effusions, and generalized edema, signs that resemble human Barth syndrome heart failure. This knockdown phenotype was rescued by concomitant injection of normal tafazzin mRNA. Abnormal cardiac development, with a linear, nonlooped heart, and hypomorphic tail and eye development proves that tafazzin is essential for overall zebrafish development, especially of the heart. The tafazzin knockdown zebrafish provides an animal model similar to Barth syndrome to analyze the severity of human mutants and to test potential treatments.

General mitochondrial trifunctional protein (TFP) deficiency as a result of either alpha- or beta-subunit mutations exhibits similar phenotypes because mutations in either subunit alter TFP complex expression and subunit turnover.

The mitochondrial trifunctional protein (TFP) is a multienzyme complex of the beta-oxidation cycle. Human TFP is an octamer composed of four alpha-subunits harboring long-chain enoyl-CoA hydratase and long-chain L-3-hydroxyacyl-CoA dehydrogenase and four beta-subunits encoding long-chain 3-ketoacyl-CoA thiolase. Mutations in either subunit may result in general TFP deficiency with reduced activity of all three enzymes. We report five new patients with alpha-subunit mutations and compare general TFP deficiency caused by alpha-subunit mutations (n = 15) to that caused by beta-subunit mutations (n = 13) with regard to clinical features, enzyme activity, mutations, thiolase expression, and thiolase protein turnover. Among patients with alpha-subunit mutations, the same three heterogeneous phenotypes reported in patients with beta-subunit mutations were observed: a lethal form with predominating cardiomyopathy; an infancy-onset, hepatic presentation; and a milder, later-onset, neuromyopathic form. Maternal HELLP syndrome (hemolysis, elevated liver enzymes, low platelets) occurred with an incidence of 15 to 20%, as in families with beta-subunit mutations. Enzyme assays in fibroblasts revealed an identical biochemical pattern in both groups. alpha-Subunit mutational analysis demonstrated molecular heterogeneity, with 53% (9 of 17) truncating mutations. In contrast, patients with beta-subunit mutations had predominantly missense mutations. Thiolase expression in fibroblasts was as markedly reduced in alpha-subunit patients as in the beta-subunit group with similarly increased thiolase degradation, presumably secondary to TFP complex instability. TFP deficiency as a result of either alpha- or beta-subunit mutations presents with similar, heterogeneous phenotypes. Both alpha- and beta-subunit mutations result in TFP complex instability, demonstrating that the mechanism of disease is the same in alpha- or beta-mutation-derived disease and explaining the biochemical and clinical similarities.