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Following oxidative stress, reactive oxygen species are produced and accumulate in glioma cells in large quantities, and to avoid the occurrence of cellular dysfunction, glioma cells can respond adaptively in the biological processes of DNA damage repair, lipid peroxidation and protein modification to produce radiotherapy resistance. The expression of nuclear factor erythroid 2-related factor 2, solute carrier family 7 member 11, glutathione and microRNA, as key regulatory molecules, can regulate reactive oxygen species levels, alter glioma oxidative stress status, and affect radiochemotherapy sensitivity. Further study on the relationship between oxidative stress and sensitivity to radiotherapy and chemotherapy of glioma can provide theoretical basis for precise treatment of glioma.
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Acute leukemia marks the main type of non-solid cancer in children.Studies have proved that long non-coding RNA plays important roles in the pathogenesis of hematological malignancies and can be applied as a biomarker for diagnosis, prognosis and efficacy monitoring of leukemia patients.This article briefly reviews the roles of long non-coding RNA in the tumorigenesis and progression of childhood acute leukemia.
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Objective:To evaluate the risk of infectious complication after endoscopic surgery for the treatment of upper urinary tract calculi combined with carbapenem-resistant Enterobacteriaceae (CRE) bacteriuria.Methods:The clinical data of 14 patients who were diagnosed with upper urinary tract calculi combined with CRE bacteriuria and treated in Tsinghua University affiliated Beijing Tsinghua Changgung Hospital from January 2015 to December 2019 were analyzed retrospectively. There were 7 males and 7 females, aged from 34 to 71 years old (mean 58.2 years old). The diagnosis was confirmed by ultrasonography, CT or abdominal X-ray. Fourteen cases underwent 15 procedures, including 4 RIRS and 11 PCNL. One patient underwent 2 PCNL procedures at an interval of 1 week, and 1 patient underwent PCNL 16 days after nephrostomy. There were 13 cases of renal calculi and 1 case of upper ureteral calculi. Stones were found on the left side in 8 cases and the right side in 6 cases. There were 3 cases of solitary stone, 4 cases of multiple stones and 7 cases of staghorn stone. The maximum diameter of stones was (31.5±10.2)mm in patients who underwent PCNL, and(10.8±2.6)mm in patients undergoing RIRS. The complete blood count, blood biochemistry, procalcitonin and C-reactive protein were tested postoperatively on the same day of the procedure and 1 day after the procedure. Abdominal X-ray was performed 1-2 days postoperatively, and the ureteral stent (double J) was removed 4 weeks after the procedure. Fourteen patients with CRE bacteriuria underwent 15 endoscopic procedures. Urine culture identified 7 cases of Escherichia coli, 6 cases of Klebsiella pneumoniae and 1 case of Enterobacter cloacae. Preoperative blood culture was performed in 3 cases, of which 1 case was negative and 1 case was Klebsiella pneumoniae positive. Before operation, 11 cases were empirical treated with broad-spectrum antibiotics, including monotherapy in 10 cases and drug combination therapy in 1 case. Sensitive antibiotics against CRE were prescribed in 4 cases preoperatively, including monotherapy in 2 cases and drug combination therapy in 2 cases. Antibiotics were used preoperatively for 1-24 days (mean 7.1 days).Results:After the operation, 7 cases received monotherapy with broad-spectrum antibiotics. Sensitive antibiotics against CRE were prescribed in 4 cases postoperatively, including monotherapy in 4 cases and drug combination in therapy 4 cases. Postoperative antibiotics were used for 2-17 days (mean 6.8 days). There were 3 cases of systemic inflammatory response syndrome (SIRS) after operation, and there were no cases of sepsis, septic shock or death. The main components of stones were ammonium magnesium phosphate hexahydrate in 8 patients and calcium oxalate monohydrate in 6 patients.Conclusions:Effective measures can be taken to reduce or avoid bacteremia caused by CRE, reducing mortality and the use of antibiotics. Endoscopic surgery can be performed only after the clinical symptoms and laboratory tests have significantly improved. Patients with fever and other clinical symptoms and abnormal infectious markers should be treated with targeted antimicrobial therapy.
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Objective@#To identify underlying reasons for discrepant cases of positive cytology but negative histology.@*Methods@#Cases with positive cytology and negative histology from 2008 to 2016 were retrieved from Department of Pathology, Obstetrics and Gynecology Hospital of Fudan University. Low grade squamous intraepithelial lesion or higher grade lesions were considered as positive cytology test in the study. Consecutive follow-up biopsies and as well as sites of biopsy were documented for analysis.@*Results@#Overall positive rate of biopsy followed positive cytology was 74.3%(8 990/12 097). Of the negative biopsies, 675 cases were followed-up with multiple biopsy. Two-hundred and eighty-seven cases (42.5%, 287/675) were confirmed to have lesions. Comparing with those with initial positive biopsiews, patients of the latter group were significantly older and had other specimen types including vaginal biopsy, cone biopsy and hysterectomy. The final histological diagnoses were well correlated with cytological results (Kappa=0.505, P<0.01).@*Conclusions@#Qualified cervical cytology is complimentary to histological diagnosis. Clinicians should not ignore the positive cytological result prior to a normal histological diagnosis. In contradictory cases, repeated colposcopy and biopsy at extended anatomic sites may reveal additional lesions.
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Sanqi in Chinese herbal medicine is the root and rhi-zoma of Panax notoginseng (Burk.)F.H.Chen. As the effects of strengthening with tonics, promoting blood circulation to re-move blood stasis,relieving swelling and pain and hemostasia,it is widely used as a tonic medicine in the traditional Chinese medicine. The main active constituents of Sanqi are panax noto-ginseng saponins,including ginsenoside Rg1,Rb1 and notogin-senoside R1. Notoginsenoside R1 is one of the unique monomer compositions of panax notoginseng,which is often used as an in-gredient indicator in new drug research and development.In cur-rent years, the scientists have been conducted tremendous fun-damental studies to research the pharmacological activities of no-toginsenoside R1, to reveal its protective effects on the cardio-vascular system,central nervous system as well as to many other aspects. It is hoped that the relevant study about the pharmaco-logical action of notoginsenoside R1 would help its further clini-cal application. This paper mainly reviews the research on the pharmacological mechanism of notoginsenoside R1 in recent years.
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A methodology of quantitative analysis on ginsenoside Re (G-Re) in rat plasma by ultra performance liquid chromatography-triple quadrupole mass spectrometry was developed for comparing the pharmacokinetic profiles between normal rats and Ultraviolet B (UVB) irradiation-induced damage rats after oral administration. The sample separation was carried out on an Ascentis?Express C18column (5.0 mm× 3.0 mm,2.7 μm) with 0.1% formic acid in water and acetonitrile as the mobile phase under gradient elution. MS analysis was operated in multiple-reaction monitoring (MRM) mode using electrospray ionization (ESI) with negative ion mode,and the ions for quantification were m/z 991.54/945.53/475.60. The limit of detection (LOD,S/N=3), limit of quantification (LOQ, S/N=10) were 4.0 ng/mL and 13.5 ng/mL, respectively. G-Re was in good linearity between 15 ng/mL and 20000 ng/mL(r=0.999),the intra-day and inter-day precisions, recovery, matrix effect and stability could meet the pharmacokinetic analysis requirement. The results indicated that the metabolic process of G-Re conformed to a two-compartment pharmacokinetic model after single oral administration in the normal and model groups. The t1/2αwere(0.21± 0.04) h and (0. 69 ± 0. 07) h, respectively; t1/2βwere (17. 08 ± 0. 53) and (21. 40 ± 16. 77) h, respectively;AUC(0-t)were (321.91±2.27) μg/(L·h) and (474.99±194.96) μg/(L·h), respectively;AUC(0-∞)were (332. 44 ± 1. 66) μg/(L·h) and (518. 64 ± 231. 39) μg/(L·h), respectively; the pharmacokinetic parameters were significantly different between normal and UVB irradiated rats (p<0.05), except for t1/2α. This UHPLC-QQQ-MS method showed excellent separation, accuracy, high sensitivity, specificity and good repeatability,and it was suitable for the pharmacokinetic study of G-Re in vivo.
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Objective :To explore therapeutic effect of single and double stent implantation on coronary bifurcation le-sions.Methods : Clinical data of 455 patients with coronary bifurcation lesions , who received drug-eluting stent (DES) implantation in General Hospital of PLA from Jan 2014 to Oct 2016 ,were retrospectively analyzed .Accord-ing to interventional strategy ,patients were divided into single stent group (n＝235) and double stent group (n＝220).The lesion distribution ,lesion features of proximal ,distal and bifurcation ,stent implantation ,surgical selec- tion ,postoperative instant blood flow ,clinical adverse events were observed and compared between two groups .Re-sults :There were no significant difference in general data ,lesion distribution suggested by coronary angiography ,le-sion feature of proximal ,distal and bifurcation between two groups , P＞0. 05 all.All patients used Cross-over tech-nique in single stent group ,while double stent group used Crush (46.81%) ,Culotte (37.73%) ,T-stent and V-stent surgery .There were no significant difference in postoperative instant TIMI blood flow grade 3 rate of main vessel and side branch between two groups , P＞0. 05 both .During hospitalization ,incidence rate of nonfatal myocardial infarction in double stent group was significantly higher than that of single stent group (7.27% vs.2.98%) , P＝0.037. During 12-month follow-up ,compared with single stent group ,there was significant reduction in restenosis rate (5.53% vs.1.36%) , and significant rise in incidence rate of nonfatal myocardial infarction (2.55% vs. 6.82%) in double stent group ,P＝0.016 ,0.030 respectively .Conclusion :The therapeutic effect between single and double stent implantation treating coronary bifurcation lesions is no significant ,but restenosis rate of double stent group significantly reduces and incidence rate of nonfatal myocardial infarction significantly rises .
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Objective To investigate the clinical efficacy of neoadjuvant chemoradiotherapy in the treatment of locally advanced low and middle rectal cancer,and evaluate the effect of related clinical factors upon the long-term survival. Methods Clinical data of 101 patients with locally advanced low and middle rectal cancer admitted to our hospital from January 1,2010 to December 31,2014 were collected. All patients completed the preoperative intensity-modulated radiation therapy DT45-50. 4 Gy,synchronized with oxaliplatin+capecitabine/5-fluorouracil or single drug capecitabine chemotherapy,and total mesorectal excision) was performed 4-13 weeks after the end of the neoadjuvant therapy. The short-term efficacy and long-term prognosis of these patients were evaluated. Kaplan-Meier method was used for survival analysis,and Cox’s regression model for multivariate analysis. Results The total sphincter preservation rate was 53. 5%.The decrease rates of T,N staging and TNM total staging were 73. 26%,67. 32% and 72. 3%,respectively. The pathological complete response ( pCR) rate was 16. 8%.The median follow-up time was 41 months. The 3-year overall survival (OS), desease-free survival (DFS),local recurrence and distant metastases rates were 82. 2%,80. 7%,7. 2% and 12. 1%,respectively. The single factor analysis demonstrated that ypT and ypN stages were the risk factors affecting the 3-year OS,DFS anddistant metastases ( all P＜0. 05).Multivariate analysis revealed that ypT stage was an independent factor affecting the 3-year OS,and ypT and ypN stages were the independent factors of the 3-year DFS ( all P＜ 0. 05 ). ConclusionsNeoadjuvant chemoradiotherapy combined with TME in the treatment of locally advanced middle and low rectal cancer can partially decrease the tumor staging,enhance the sphincter preservation rate and improve long-term clinical prognosis. Both ypT and ypN stages are correlated with the clinical prognosis of patients.
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A method for simultaneous detection of multi-mycotoxins in ten kinds of common dried fruits was been exploited by using ultra performance liquid chromatography-tandem mass spectrometry. The edible part of dried fruits was ground up with certain quantity of water. The mash was extracted by 10 mmol/L citric acid-acetonitrile, purified by C18 column, and filtered using 0. 22-μm organic filtration. All the sixteen mycotoxins were well separated in 8 min on ACQUITY UPLC BEH C18 column with acetonitrile as phase A and 10 mmol/L ammonium acetate solution containing 0. 1% formic acid as mobile phase B. Result showed that the limits of detection ( LODs) for the sixteen mycotoxins were 0. 01-1. 00 μg/L, with linearity range of 1-200 μg/L and correlation coefficients above 0 . 9981 . The average recoveries of the sixteen mycotoxins in ten matrices were 70. 48% -118. 85%, and the relative standard deviation ( RSD, n=6 ) was 0. 3% -11. 9%. This economical, fast, simple and efficient method could be used for simultaneous detection of multi-mycotoxins in different dried fruits matrixes.
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Hepatitis B virus (HBV) infection has been one of the most important public health problems,however,no complete cure is currently available.Although interferon (IFN)-α has been clinically used as a drug for chronic hepatitis B therapy because of its advantages including a higher rate of HBsAg/HBeAg seroconversion and a lower rate of recurrence after cessation of treatment,only 20％ -40 ％ of patients respond well to IFN therapy,thus hampering its clinical application.In recent years,based on the in vitro HBV replication and infection cell models,animal models and patient cohort with hepatitis B and by using a variety of methods,studies have been made.On the one hand,to identify new mechanisms underlying the IFN-and IFN-induced genes-mediated anti-HBV activities and signaling transduction,on the other hand,to reveal the effect and mechanisms of HBV replication and viral proteins in regulating the innate immune signaling pathways and IFN induction and antiviral action,based on which new strategies and approaches for optimization of IFN-based therapy and for a HBV cure have been further explored.This review mainly introduces the research findings of author's group and the future development is prospected.
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Aim To verify the mechanisms underlying the protective effects of the 4-hydroxybenzaldehyde (4-HBd)on cerebral ischemia / reperfusion in vivo and in vitro. Methods The model of focal cerebral ischemi-a / reperfusion in rats was established by the suture-oc-clusion method. After reperfusion,neurological score and cerebral infarction rate were assessed. Meanwhile, after pretreatment with 4-HBd for 24 h,the primary cultured cortical neuron cells were exposed to oxygen glucose deprivation / reoxygenation (OGD/ Rep). The survival rate, MDA, SOD, NO, Bax, Bcl-2 and caspase-3 concentrations were assessed. Results Pre-treatment with 4-HBd significantly decreased neurologi-cal score (P < 0. 05)and cerebral infarction rate (P <0. 01)in the cerebrum after MCAO/ R injury. Further-more,4-HBd significantly increased cell survival rate (P < 0. 05),decreased MDA content (P < 0. 05),in-creased SOD activity (P < 0. 05),decreased NO levels (P < 0. 05),decreased caspase-3 (P < 0. 05),in-creased Bcl-2 (P < 0. 05)but decreased Bax (P <0. 05)levels and therefore increased Bcl-2 / Bax ratio in the cerebrum after OGD/ R injury. Conclusions 4-HBd has significant protective effect on MCAO/ R rats, and the mechanism may be related to decreasing NO levels,Bax,Bcl-2,caspase-3 and reducing neuronal apoptosis.
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Objective To evaluate the role of new strategy in the transmission control of schistosomiasis in Poyang Lake re-gion. Methods The information and epidemic data of schistosomiasis control were collected and analyzed in Poyang Lake re-gion from 2005 to 2016. Results After eleven years of carrying out the new strategy,thirteen counties achieved the objective of transmission control in Poyang Lake region. In 2016,the number of schistosomiasis cases and human infection rate were 10301 and 0.03％,decreased by 89.64％and 99.45％compared with those in 2005,respectively. The number of cattle and schistosome-infected cattle were 68152 and 5,decreased by 50.84％and 99.83％compared with those in 2005,respectively. The average density of Oncomelania hupensis snails was decreased by 61.52％. No schistosome-infected snails were found since 2014. Con-clusion The new strategy accurately locates the key points and targets of schistosomiasis transmission chain ,which has con-trolled the human and animal's fecal eggs from polluting grassland,and cut off the transmission chain,reduced both the infec-tion rates of human and animal and the re-infection risk,and promoted to achieve the target of schistosomiasis transmission con-trol in Poyang Lake region.
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Objective To develop a new mPCR method for rapid diagnosis of six types of encephalitis causing viruses of HS-VI,HSVII,VZV,EBV,EV71 and CMV.Methods Six pairs of specific primers for CMV,EV71,HSV I,VZV,EBV and HSV II were designed.The mPCR detection method was established and the sensitivity was detected.In order to verify the clinical application value of their multiplex PCR system,fifteen cerebrospinal fluid specimens of clinically suspected VE from the First Affiliated Hospital of Soochow University from 2014 to 2015 were examined by the mPCR method.Results The 6 pairs of primers did not interfere with each other,and the sensitivity of the mPCR system was over 103copies/μl.Among 15 cerebrospinal fluid specimens from patients with suspected viral encephalitis,six specimens(6/15,40%)were tested positive by the mPCR.Among them,HSV I was 5 and CMV was 1.Conclusion The mPCR method for detecting six types of en-cephalitis-associated virus at same time was established with high specificity,sensitivity and stability.
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Objective: To explore the safety and efficacy of a novel lead locking device (LLD) in the procedure of cardiac lead extraction for heart rhythm implants. Methods: A total of 6 patients using LLD for cardiac lead extraction in our hospital were retrospectively reviewed. Clinical parameters, the reason of cardiac lead extraction, lead locking stylet condition, outcome of lead extraction and operative complications were summarized. Results: There were 6 patients including 1 female with the median age at 62.5 years. LLD was used and 13 cardiac leads were extracted including 1 scrap electrode wire and 12 functional electrode wire. Among those, LLD was successfully inserted and locked on the top of 11/13 (85%) leads for whole procedure and 2 (15%) leads were not locked for whole procedure; 12 (92 %) leads were completely removed and 1 (8%) lead was partially removed. No severe complications occurred. Conclusion: The novel LLD may safely and effectively extract electrode lead which is beneficial for complete cardiac lead extraction.
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A method for simultaneous detection of multi-mycotoxins in ten kinds of common dried fruits was been exploited by using ultra performance liquid chromatography-tandem mass spectrometry. The edible part of dried fruits was ground up with certain quantity of water. The mash was extracted by 10 mmol/L citric acid-acetonitrile, purified by C18 column, and filtered using 0. 22-μm organic filtration. All the sixteen mycotoxins were well separated in 8 min on ACQUITY UPLC BEH C18 column with acetonitrile as phase A and 10 mmol/L ammonium acetate solution containing 0. 1% formic acid as mobile phase B. Result showed that the limits of detection ( LODs) for the sixteen mycotoxins were 0. 01-1. 00 μg/L, with linearity range of 1-200 μg/L and correlation coefficients above 0 . 9981 . The average recoveries of the sixteen mycotoxins in ten matrices were 70. 48% -118. 85%, and the relative standard deviation ( RSD, n=6 ) was 0. 3% -11. 9%. This economical, fast, simple and efficient method could be used for simultaneous detection of multi-mycotoxins in different dried fruits matrixes.
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<p><b>OBJECTIVE</b>To determine the optimum treatment for viral myocarditis (VMC).</p><p><b>METHODS</b>A total of 126 VMC patients were randomly divided into the control group (42 cases) that was treated with conventional Western medicine, and the intervention group (84 cases) that was treated with a combination of Chinese medicine (CM) and Western medicine intervention termed optimum proposal of integration of disease and syndrome (OPIDS). Before and after 4 weeks of treatment, the integral of CM syndrome, self-rating depression and anxiety scales (SDS and SAS, respectively), echocardiograms (ECGs), heart rate variability and left ventricular systolic function were observed.</p><p><b>RESULTS</b>Compared with the control group, the intervention group showed significant reductions on the SDS and SAS (P <0.05); improvement of premature ventricular beats, atrioventricular blocks, ST-segment abnormalities, and significant T wave changes (P <0.05); greater reductions in standard deviation of NN intervals (SDNN), standard deviation for per 5 min averages NN intervals (SDANN), and root-mean-square of successive difference of NN intervals (rMSSD) (P <0.05); and increases in cardiac output, stroke volume, and ejection fraction, the last of which was statistically significant (P <0.05). Overall, the treatment efficacy rate was significantly better P<0.05) in the intervention group (75.61%) compared with the control group (69.70%).</p><p><b>CONCLUSION</b>OPIDS is quite effective in treating VMC and improves symptoms such as anxiety and depression, left ventricular systolic dysfunction, premature ventricular contraction, and cardiac autonomic nervous system dysfunction. [</p><p><b>REGISTRATION</b>Chinese clinical trial center (No. ChiCTR-TRC-00000298)].</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Young Adult , Anxiety , Depression , Electrocardiography , Heart Rate , Medicine, Chinese Traditional , Myocarditis , Diagnostic Imaging , Therapeutics , Virology , Syndrome , Systole , Ultrasonography , Ventricular FunctionABSTRACT
Background Corneal epithelial abrasion results in corneal ulcer and stroma cloudy evenb irreversible visual impairment.Previous drugs for corneal epithelial injury can only alleviate the inflammatory irritation.So it is very important to seek a drug which regulate the growth of corneal epithelium.Objective This study was to investigate the effects of recombinant human BIGH3 protein eye drops on corneal epithelial abrasion.Methods Fifty right eyes of 50 clean adult New Zealand white rabbits were collected.Two rabbits were sacrificed right away following establishment of corneal epithelial abrasion models (0 hour group).The other 48 rabbits were randomly divided into recombinant human epidermal growth factor (EGF) derivative group (positive control group),normal saline solution group (negative control group),0.25％ or 0.5％ recombinant human BIGH3 protein eye drops group.Corneal abrasion models were created with alcohol corrosion method with a defect area of 7 mm2.The corresponding eye drops were used separately in 4 groups for four times per day after operation.Experimental eyes were examined by the slit lamp microscope,and fluorescein vital staining were performed 12,24,36,48,72 hours after operation.Planimetry was performed and the corneal photographs were analyzed with computer software.The rabbits were sacrificed 12,24,36,48 and 72 hours after operation,respectively,and the histopathological examination of corneal tissue was carried out.Results No obvious irritation response was seen after administered of eye drops in the recombinant human EGF derivative group,normal saline solution group,0.25％ and 0.5％ recombinant human BIGH3 protein eye drops groups.Histopathological examination revealed a full-thickness defect of corneal epithelium after modeling.The defect area was gradually smaller with time lapse,and corneal epithelium migrated from periphery toward the center zone.Corneal epithelial cells increased with time lapse.Compared with normal saline solution group,the defect area of corneal epithelium lessened 12,24,36,48 hours after operation in the 0.25％,0.5％ recombinant human BIGH3 protein eye drops groups and recombinant human EGF derivative group (all at P =0.000),but at 12and 24,36 hours after operation,no significant differences were found between the recombinant human EGF derivative group and normal saline solution group (P =0.321,0.057,0.126).The defect area was smaller in the 0.5％recombinant human BIGH3 protein eye drops group than that of the recombinant human EGF derivative group at various time points (P=0.042,0.039,0.025,0.008).However,significant smaller defect area was exhibited only at 12 hours and 24 hours after operation in the 0.25％ recombinant human BIGH3 protein eye drops group (P=0.047,0.042).No significant differences were seen in corneal defect area at various time points between 0.25％ and 0.5％recombinant human BIGH3 protein eye drops groups (P =0.358,0.259,0.108,0.062).In addition,the corneal defect area was (0.51 ±0.42)mm2 72 hours after operation in the normal saline group;while that in the recombinant human EGF derivative group and recombinant human BIGH3 protein eye drops groups was disappeared.The repairing curves in the recombinant human BIGH3 protein eye drops groups were superior to those of the recombinant human EGF derivative group and normal saline solution group.Conclusions 0.25％ and 0.5％ recombinant human BIGH3 protein eye drops have facilitation effect on the growth of corneal epithelial cells and the healing of corneal injury.
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<p><b>OBJECTIVE</b>To further select the items based on the pre-test version of quality of life scale in patients with viral myocarditis.</p><p><b>METHODS</b>Totally 100 patients with viral myocarditis were enrolled in this study. Methodologies including frequency distribution, discrete trend, t-test, Cronbach's α coefficient, correlation coefficient and factor analysis were applied to select items from different perspectives.</p><p><b>RESULTS</b>A total of 17 items were selected by frequency distribution method from the perspective of central tendency, 15 items were selected by discrete trend method from the perspective of sensitivity, 16 items were selected by t-test method from the perspective of sensitivity and discrimination, 16 items were selected by Cronbach's α coefficient method from the perspective of internal consistency, 12 items were selected by correlation coefficient method from the perspective of representation and independence, and 18 items were selected by factor analysis method from the perspective of representation.</p><p><b>CONCLUSION</b>Item selection of quality of life scale in patients with viral myocarditis was successfully conducted based on the clinical epidemiological data using a variety of statistical methods.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Young Adult , Myocarditis , Virology , Quality of Life , Surveys and QuestionnairesABSTRACT
BackgroundEndostatin (ES) is currently the strongest endogenous angiognesis inhibitor,and it can inhibit the occurrence of neovascularization.Various studies demonstrated that the poly RGD sequence can enhance the function of the ES gene.ObjectiveThis study was to evaluate the use of gene therapy of modified ES for alkaline burn-induced corneal neovascularization (CNV).MethodsOne hundred and two clean SD rats were randomly divided into the normal control group,the pCI empty vector group,the pCI-ES group,and the pCI-RGDRGDES group.Corneal neovascularization models were established by placing a piece of 3 mm filter paper with 1 mol/L NaOH at the central cornea for 40 seconds.3 μg of the pCI blank vector,ES-tranfected pCI blank vector,or RGDRGD-ES-transfected pCI vector was injected into the superior bulbar conjunctiva after the alkali burn twice at 1-week intervals.Area of CNV and edema of the cornea in the various groups of rats were examined daily under the slit lamp biomicroscope.1,4,7 and 14 days after operation,the rats were sacrificed by the excessive anesthesia method and corneal tissues were obtained to evaluate pathological changes.The expression of CD34 in vascular endothelial cells was detected by immunochemistry to calculate the corneal neovascular density.The expressions of VEGF mRNA and Flk-1 protein in the corneas were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.The use and maintenance of animals followed the Statement of ARVO.Results Seven to fourteen days after corneal alkali-burning,the corneal neovascular area was smaller in the pCI-ES group and pCI-RGDRGD-ES group compared with the normal control group and pCI blank vector group (P＜0.05,P＜0.01 ),and nevascular area in the pCI-RGDRGD-ES group was smaller than that in the pCI-ES group (P＜0.05).The expression level of CD34 was significantly lower in the pCI-ES group and pCI-RGDRGD-ES group than that in the normal control group and pCI blank vector group (P＜0.05,P＜0.01 ),and the expression level of CD34 was further declined in the pCI-RGDRGD-ES group compared with the pCI-ES group (P＜0.05 ).Compared with the normal control group and pCI vector group,the expressions of the Flk-1 protein and VEGF mRNA were decreased in the pCI-ES group and pCI-RGDRGD-ES group on the fourth day after corneal alkali-burning (P＜0.01,P＜0.05 ),and those in the pCI-RGDRGD-ES group were less than the pCI-ES group (P＜ 0.05,P＜ 0.05 ).Conclusions Subconjunctival injection of both ES and modified RGDRGD-ES genes result in significant suppression of CNV in vivo,and modified RGDRGD-ES appears to be more effective than native ES.The main mechanism of ES in inhibiting neovascularization is to downregulate the expression of VEGF and Flk-1.
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BACKGROUND: Horse serum (HS) was a strong allergen for xenogeneic animals. However, the methods of test for allergen didn't authorized in Pharmacopeia of the United States, European Pharmacopeia, Japanese Pharmacopeia and British Pharmacopeia. Thus, new methods of test for allergen are required to control drug allergy. OBJECTIVE: To propose a new method for detecting horse serum induced allergic reactions of guinea pigs earlier. METHODS: Guinea pigs were sensitized successively by injecting different concentration of HS intravenously once a day for three times, serum level of IL-4 and total IgE were detected by method of enzyme linked immunosorbent assay (ELISA) before guinea pigs were challenged by injecting HS intravenously only once, and the results were compared with routine method of sensitization by injecting HS intraperitoneally every other day for three times. RESULTS: Serum level of IL-4 and total IgE increased significantly before guinea pigs were challenged, either in day 8 after intravenous sensitization (10%HS, 0.5 mL) or in day 14 and day 21 after intraperitoneal sensitization (10%HS, 0.5 mL), and allergic reactions occurred in all guinea pigs after challenged by injecting HS (10%, 1.0 mL) only once. CONCLUSION: It provides a new way to predict whether HS or other drugs can provoke allergic reactions earlier by detecting the serum level of IL-4 and total IgE in d8 after intravenous sensitization, this has a good application value in drug emergency test.