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Objective:To evaluate the value of a three-dimensional inversion-recovery with real reconstruction (3D-real IR) sequence with an ultralong repetition time (TR) for the endolymphatic hydrops (EH) of Meniere disease (MD) after intravenous gadolinium administration, and compare it with a heavily T 2-weighted three-dimensional fluid-attenuated inversion recovery (hT 2-3D-FLAIR) sequence. Methods:From July 2021 to July 2022, 52 definite MD patients (58 ears) were retrospectively enrolled at Zhongshan Hospital, Fudan University. The 3D-real IR with an ultralong TR (16 000 ms) and hT 2-3D-FLAIR sequences were performed four hours after intravenous single-dose gadolinium administration. The image quality of the two sequences was rated. The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were measured in the two sequence. The EH of cochlear and vestibular was graded, and EH detection rates were calculated. Scores of the two sequences were compared using the paired Wilcoxon signed rank test. Paired t test was used to compare the differences of the SNR and CNR. McNemar test was performed to compare the EH detection rate between the two sequences. Results:The score of the 3D-real IR [3 (3, 4)] was significantly higher than that of the hT 2-3D-FLAIR [2.5 (2, 3), Z=-6.06, P<0.001]. No significant difference was found in SNR of 3D-real IR and hT 2-3D-FLAIR (11.4±6.5 and 12.3±3.7, t=-1.38, P=0.175). CNR of the 3D-real IR (21.7±9.3) was significantly higher than that of the hT 2-3D-FLAIR (9.7±3.8, t=10.67, P<0.001). Using 3D-real IR sequence, the EH detection rate of cochlear (89.7%, 52/58) was higher than using hT 2-3D-FLAIR (67.2%, 39/58, χ 2=11.10, P<0.001). No significant difference was found in the EH detection rate of vestibular between 3D-real IR (77.6%, 45/58) and hT 2-3D-FLAIR (74.1%, 43/58, χ 2=0.50, P=0.500). Conclusion:Compared with hT 2-3D-FLAIR sequence, the 3D-real IR with an ultralong TR can improve the depiction of EH in MD patients after intravenous single-dose gadolinium administration. It can provide higher image quality and detection rate of EH.
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Retinal pigment epithelial (RPE) is primarily impaired in age-related macular degeneration (AMD), leading to progressive loss of photoreceptors and sometimes choroidal neovascularization (CNV). mTOR has been proposed as a promising therapeutic target, while the usage of its specific inhibitor, rapamycin, was greatly limited. To mediate the mTOR pathway in the retina by a noninvasive approach, we developed novel biomimetic nanocomplexes where rapamycin-loaded nanoparticles were coated with cell membrane derived from macrophages (termed as MRaNPs). Taking advantage of the macrophage-inherited property, intravenous injection of MRaNPs exhibited significantly enhanced accumulation in the CNV lesions, thereby increasing the local concentration of rapamycin. Consequently, MRaNPs effectively downregulated the mTOR pathway and attenuate angiogenesis in the eye. Particularly, MRaNPs also efficiently activated autophagy in the RPE, which was acknowledged to rescue RPE in response to deleterious stimuli. Overall, we design and prepare macrophage-disguised rapamycin nanocarriers and demonstrate the therapeutic advantages of employing biomimetic cell membrane materials for treatment of AMD.
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Objective@#To investigate the protective effect of human umbilical cord mesenchymal stem cells (UCMSCs) on light-damaged retinal pigment epithelial (RPE) cells in vitro.@*Methods@#Human UCMSCs were cultured, and then identified by flow cytometry.Human RPE cells were isolated and cultured, and then the model of light-damaged RPE cells was prepared.The noncontact co-culture system of light-damaged RPE cells and UCMSCs was established by Transwell chamber.RPE cells were divided into normal control group, model control group and UCMSCs co-culture group.RPE cells in the normal control group were not treated.RPE cells in the model control group were treated with blue light to induce RPE cell damage.Co-culture system of light-damaged RPE cells and UCMSCs was established in the UCMSCs co-culture group.The proliferative ability of RPE cells was measured by methyl thiazolyl tetrazolium (MTT) assay at 24 hours and 48 hours after co-culture.ELISA kits were used to quantitatively measure the levels of pigment epithelium-derived factor (PEDF) and basic fibroblast growth factor (bFGF) in the culture supernatant at 48 hours after co-culture.And the photoreceptor outer segments (POS) phagocytosis assay of RPE cells was also conducted.@*Results@#UCMSCs displayed spindle-shaped morphology, positively expressed CD29, CD44, CD90 and CD105, while negatively expressed CD34 and CD45.RPE cells were polygonal in morphology and positive for the specific marker RPE65 protein.The proliferative ability(A value) of RPE cells in the three groups at different timepoints were significantly different (Fgroup=132.388, P=0.000; Ftime=231.440, P=0.000), the A values of RPE cells in model control group and UCMSCs co-culture group were significantly lower than that in the normal control group, the A value of RPE cells in UCMSCs co-culture group was significantly higher than that in the model control group, and the differences were statistically significant both at 24 hours and 48 hours after co-culture (all at P<0.01). POS phagocytosis test showed that there was a significant difference in the average number of POS particles phagocytized by RPE cells among the three groups(F=28.087, P=0.000). The average number of POS particles phagocytized by RPE cells in model control group was significantly lower than that in normal control group, and the average number of POS particles phagocytized by RPE cells in UCMSCs co-culture group was significantly more than that in model control group (all at P<0.01). ELISA showed that the concentrations of PEDF in RPE cell supernatant of normal control group, model control group and UCMSCs co-culture group were (18.8±1.9), (10.0±1.7) and (20.2±6.0)ng/ml, respectively.The concentrations of bFGF in RPE cell supernatant were (25.2±1.5), (26.3±3.6) and (61.9±14.3)pg/ml, respectively.There were significant differences in PEDF and bFGF concentrations among the three groups (F=8.654, P=0.008; F=23.698, P=0.000). The concentration of PEDF in the model control group was significantly lower than that in the normal control group, and the concentration of PEDF in the UCMSCs co-culture group was significantly higher than that in the model control group (all at P<0.01). The concentration of bFGF in UCMSCs co-culture group was significantly higher than that in the normal control group and model control group (all at P<0.01).@*Conclusions@#Cocultivation with UCMSCs can improve the proliferation ability and phagocytosis function of light-damaged RPE cells, and promote the secretion of PEDF by RPE cells.UCMSCs may protect RPE cells from light damage through paracrine secretion of bFGF.
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Objective To investigate the basic situation of ultrasound departments in secondary and tertiary public hospitals in Anhui Province.Methods Ultrasonic quality baseline survey forms were issued by the Ultrasonic Medical Quality Control Center of Anhui Province in November 2018,and the subjects of the survey were ultrasound departments of public secondary and tertiary hospitals in Anhui Province.The survey consisted of six parts,including the basic information of the head of ultrasound department,the number of staff,the area of ultrasound department,the number of ultrasonic diagnostic instrument rooms,the number of ultrasonic diagnostic instruments,and the number of beds actually opened in the hospital.Results In this survey,the results of 205 hospitals in 15 prefectures and cities in Anhui Province were received.Among them,there were 62 tertiary hospitals and 143 secondary hospitals.Most of the heads of ultrasound departments in Anhui public secondary and tertiary hospitals had a bachelor's degree,a middle or senior professional title,and a professional life of more than 20 years.The average number of ultrasound department staff was 9.3 in the secondary hospitals and 19.3 in the tertiary hospitals.The average area of the ultrasound department was 195.1m2 in the secondary hospitals and 505.9 m2 in the tertiary hospitals.The average number of ultrasonic diagnostic instrument rooms was 4.9 in the secondary hospitals and 11.3 in the tertiary hospitals.The average number of ultrasound instruments was 4.9 in the secondary hospitals and 11.5 in the tertiary hospitals.The average number of beds actually opened was 510.7 in the secondary hospitals and 1407.5 in the tertiary hospitals.Conclusions At present,ultrasound instruments in secondary hospitals in Anhui Province can basically meet the needs of clinical work,and there is a relative shortage of ultrasound instruments in tertiary hospitals.The shortage of staff in ultrasound department is a common problem faced by secondary and tertiary hospitals,especially tertiary hospitals.
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Objective To investigate the genetic basis of patients with intellectual disability,and to assess the application of single nucleotide polymorphisms (SNP)-array in the molecular diagnosis of intellectual disability.Methods Sixty-four patients with intellectual disability who were identified in Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region from January 2013 to June of 2015 were enrolled.Genomic DNA was extracted from peripheral blood and was analyzed with Illumina Humancyto SNP-12 300K gene array chip.All identified copy number variants (CNVs) were analyzed with references from databases such as ClinVar,DECIPHER,OMIM and DGV(Database of Genomic Variants),as well as comprehensive literature review from PubMed database to determine the pathogenicity of CNVs.Results Sixteen cases of the above 64 patients were found to have CNVs with genomic alterations,including 6 cases microdeletions/microduplications associated with known syndromes,3 cases microdeletions and microduplications with clear clinical relevance (non-syndrome),1 case numerical chromosome aberration,1 case unbalanced translocation and 5 cases CNVs of unknown clinical significance.The detection rate was 25% (16/64 cases).Among these 16 abnormalities,6 cases of them could not be detected by using karyotyping analysis because their sizes were less than 5 Mb,and the smallest detected missing fragment was 0.53 Mb.Conclusion SNP-array gene chip technique with the advantages of higher efficiency,high-resolution and good accuracy,which can be applied to the genetic diagnosis of intellectual disability.
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Objective To analyze MR imaging appearance of pancreatic neuroendocrine neoplasms (pNEN) and to investigate the practical value of MR imaging appearance in predicting pathological grade of pNEN. Methods All data of 35 patients with pathologically proven pNEN were retrospectively reviewed. MR examinations of the abdomen were performed on all these patients before operation and the data of clinical, pathological and MR imaging were intact. Both plain scan and contrast-enhanced MR scan were performed on each patient. Histopathological grade of pNEN was defined as: G1, G2, and G3 according to World Health Organization classification of tumours of the digestive system guidelines(2010). Image analysis included tumor location, number, size, shape, lesion margins, signal intensity, enhancement pattern, main pancreatic duct dilatation, extrapancreatic spread, and metastases of lymph node and liver. The comparison of quantitative index between G1 and G2 group was performed with t test. Categorical variables were tested using Fisher exact test. Results Thirty five lesions were found in 35 patients, with 14 lesions in G1, 19 lesions in G2, and 2 lesions in G3.Thirty three lesions appeared as a solid mass, and 2 lesions appeared as a cystic lesion. Significant gender-based difference was found between G1 group and G2 group (P0.05). Of the 35 lesions, 27 lesions were round in shape, while other 8 lesions were irregular. There were 18 lesions with clear margin, and the margins in other 17 lesions were blurred. Main pancreatic duct dilatation was found in 3 cases(1 in G2, 2 in G3). Significant differences in tumor diameter, shape, margin, signal intensity on precontrast images, extrapancreatic spread and metastases were found between G1 group and G2 group(P<0.05). No significant difference was found in main pancreatic duct dilatation or signal intensity on all enhancement phases between G1 group and G2 group. The 2 lesions in G3 group appeared mild contrast enhancement with degrees lower than the pancreas in all enhancement phases. Conclusion MR imaging features such as tumor diameter, shape, margin, signal intensity on precontrast images, extrapancreatic spread and metastases may preoperatively predict the pathological graden of pNEN.
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<p><b>OBJECTIVE</b>To explore the value of single nucleotide polymorphism array (SNP-array) for the analysis of pediatric patients with growth retardation.</p><p><b>METHODS</b>One hundred eighty one children with growth retardation were enrolled. DNA was extracted from peripheral samples from the patients, and whole genome copy number variations (CNVs) were detected using Illumina Human Cyto SNP-12. All identified CNVs were further analyzed with reference to databases including ClinGen, ClinVar, DECIPHER, OMIM and DGV as well as comprehensive review of literature from PubMed to determine their pathogenicity.</p><p><b>RESULTS</b>Forty seven patients (26%) with abnormal CNVs were detected, which included 12 known microdeletions/microduplications syndrome (26%), 10 pathogenic non-syndromic CNVs (21%), 3 numerical chromosome aberrations (6%), 3 unbalanced translocations (6%), 4 pathogenic mosaicisms (9%) and 15 cases with unknown clinical significance (32%). After excluding obvious numerical and/or structural chromosomal abnormalities, this study has detected 15 pathogenic microdeletions/microduplications sized 5 Mb or less, which may be missed by routine chromosomal karyotyping. In addition, there were 3 cases with loss of heterozygoisty (LOH) containing known or predicted imprinting genes as well as 2 cases with suspected parental consanguinity.</p><p><b>CONCLUSION</b>SNP-array technology is a powerful tool for the genetic diagnosis of children with growth disorders with advantages of high resolution and improved accuracy.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Male , Chromosome Aberrations , DNA Copy Number Variations , Developmental Disabilities , Diagnosis , Genetics , Karyotyping , Oligonucleotide Array Sequence Analysis , Methods , Polymorphism, Single NucleotideABSTRACT
<p><b>OBJECTIVE</b>To explore the molecular mechanism for a boy suspected with 3-methylcrotonyl-CoA carboxylase deficiency by neonatal screening.</p><p><b>METHODS</b>PCR and Sanger sequencing were used to identify potential mutations of MCCC1 and MCCC2 genes. SIFT and Polyphen-2 software was used to predict the effect of variant on the protein function and conservation of the variant across various species. Human Splicing Finder and Swiss-PdbViewer4.1.0 were applied to analyze the possible mechanism of the variant.</p><p><b>RESULTS</b>For the proband, a compound heterozygous mutation was discovered in the MCCC1 gene, namely c.539G>T (p.G180V) and c.704_711del (p.A235Vfs*4), which were inherited from his father and mother, respectively. The two mutations have disrupted the protein conformation, which in turn may impact the function of MCC protein.</p><p><b>CONCLUSION</b>The compound heterozygous mutations of the MCCC1 gene may contribute to the 3-methylcrotonyl-CoA carboxylase deficiency manifested by the patient.</p>
Subject(s)
Humans , Infant, Newborn , Male , Amino Acid Sequence , Base Sequence , Carbon-Carbon Ligases , Chemistry , Genetics , DNA Mutational Analysis , Heterozygote , Models, Molecular , Mutation , Neonatal Screening , Methods , Protein Conformation , Sequence Homology, Amino Acid , Urea Cycle Disorders, Inborn , Diagnosis , GeneticsABSTRACT
Objective To evaluate the role of c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38MAPK) signaling pathways in reduction of ischemia-reperfusion (I/R) injury by morphine preconditioning in the rats with heart failure.Methods Adult male Sprague-Dawley rats,weighing 200-230 g,in which doxorubicin 2 mg/kg was injected via the tail vein once a week for 6 consecutive weeks to induce chronic heart failure,were studied.At the end of 8th week,45 rats with chronic heart failure were randomly divided into 5 groups (n =9 each) using a random number table:sham operation group (group S),group I/R,morphine preconditioning group (group MPC),SP600125 (JNK inhibitor) + morphine preconditioning group (group MSP) and SB203580 (p38MAPK inhibitor) + morphine preconditioning group (group MSB).Myocardial I/R was induced by 30 min occlusion of the anterior descending branch of the coronary artery followed by 120 min reperfusion in each group except group S.In group MPC,the rats were subjected to 3 cycles of 5-min infusion of 0.1 mg/kg morphine via the femoral vein at 5 min interval before ischemia.In MSP and MSB groups,SP600125 0.5 mg/kg and SB203580 0.2 mg/kg were injected via the femoral vein,respectively,at 10 min before morphine preconditioning.The animals were sacrificed at 120 min of reperfusion,and the myocardial specimens were obtained for determination of the total areas of right and left ventricles (LV+RV),area at risk (AAR),infarct size (IS),and expression of PKC δ in myocardial tissues (by immunohistochemistry),and IS/AAR ratio was calculated.Results There was no significant difference in LV+RV and AAR between the five groups (P＞0.05).Compared with group S,IS and IS/AAR were significantly increased,and the expression of PKC δ was upregulated in I/R and MSB groups (P＜0.05).Compared with group I/R,IS and IS/AAR were significantly decreased,and the expression of PKC δ was down-regulated in MPC and MSP groups (P＜0.05).Compared with group MPC,IS and IS/AAR were significantly increased,and the expression of PKC δ was upregulated in group MSB (P＜0.05),and no significant change was found in the parameters mentioned above in group MSP (P＞0.05).Conclusion Activation of p38MAPK signaling pathway is involved in reduction of myocardial I/R injury by morphine preconditioning,and the mechanism is related to down-regulation of PKC δ expression in rats with heart failure;JNK signaling pathway is not involved in this process.
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<p><b>OBJECTIVE</b>To explore the molecular etiology for a Chinese family affected with isolated methylmalonic acidemia (MMA).</p><p><b>METHODS</b>Potential mutations of MUT, MMAA and MMAB genes in the proband were screened by PCR and Sanger sequencing. The pathogenicity of identified mutations was analyzed using Polyphen2, SIFT, HSF, DNAMAN 6.0 and Swiss-PdbViewer4.1.0 software.</p><p><b>RESULTS</b>Two novel mutations of the MUT gene, including c.581C>T (p.P194L) and c.1219A>T (p.N407Y), were discovered in the proband, which were inherited respectively from his mother and father. Bioinformatics analysis suggested that both mutations were damaging. The affected codons P194 and N407, both located in the (beta, alpha) 8 barrel domain and to which the substrate methylmalonyl-CoA is bound, are highly conserved across various species. Both mutations can disrupt the space conformation of its protein product, affecting the function of the MCM protein.</p><p><b>CONCLUSION</b>The novel mutations of MUT gene probably underlie the isolated MMA in this family.</p>
Subject(s)
Adult , Animals , Female , Humans , Infant , Male , Amino Acid Metabolism, Inborn Errors , Genetics , Amino Acid Sequence , Asian People , Genetics , Base Sequence , China , Methylmalonyl-CoA Mutase , Genetics , Molecular Sequence Data , Mutation , Mutation, Missense , Pedigree , Point Mutation , Sequence AlignmentABSTRACT
Objective To investigate the value of syngo WARP technology for spine MR imaging in patients with metallic prosthesis.Methods Twenty-five patients with metallic prosthesis in cervical spine or lumbar spine were prospectively included in this study.Both conventional MR sequences and optimized sequences with syngo WARP were applied in all patients.Acquisition time and image quality of two sequences were compared using a paired t test.Results All patients were examined successfully.Scanning time of cervical spine was 8 min 16 s vs.12 min 45 s for conventional sequences and syngo WARP optimized sequences respectively,with a significant difference (t =7.963,P ＜ 0.01).Scanning time of lumbar spine was longer by syngo WARP optimized sequences compared with conventional sequences (11 min 53 s vs.9 min 16 s),with a significant difference (t =4.904,P ＜ 0.01).However,image quality was better for syngo WARP optimized sequences compared with conventional sequences (4.22 ± 0.67 vs.3.56 ± 0.53 ;t =3.364,P =0.002).STIR with syngo WARP could optimize the signal loss of metallic prosthesis surrounding tissues,and significantly improve image distortion and blurring compared to the conventional sequence.Conclusion Syngo WARP technique could effectively reduce metal artifacts and better display metal implant surrounding tissue and anatomical structure with potential clinical value.
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Objective To establish a quality standard for Aiweixin oral liquid. Methods TLC was applied to the identification of Caryophylli Flos and Crocin in Aiweixin oral liquid. Eugenol was analyzed by GC. Results TLC of Caryophylli Flos and Crocin had distinct separation of characteristic spots and there was no interference in negative comparison. The linear response ranges of Eugenol were between 0.052 17-2.086 8μg (r=0.999 9). The samples were steady within 24 h. The average recovery of eugenol was 95.7%, RSD=1.4% (n=6). Conclusion The method is simple, accurate and reliable, and can be used for the quality control of Aiweixin oral liquid.
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Aim To investigate the roles of mitogen-ac-tivated protein kinases ( MAPK ) pathways in the pro-tective effects of remifentanil preconditioning against is-chemia/reperfusion injury of isolated heart in rats with heart failure. Methods Adult male SD rats were injected with adriamycin via tail vein for 6 weeks to induce heart failure. The rats were confirmed chronic heart failure through echocardiography and randomly divided into 9 groups(n=6)as follows: sham group, ischemia/reperfusion group ( IR) , remifentanil precon-ditioning group( RPC) , ERK inhibitor PD98059+RPC group ( RPD ) , p38 inhibitor SB203580 +RPC group ( RSB ) , JNK inhibitor SP600125 + RPC group ( RSP ) , and the inhibitor control groups ( PD , SB and SP) . All hearts were linked to the Langendorff ap-paratus. The coronary effluent was collected to detect the activity of lactate dehydrogenase ( LDH ) at base-line, 5 min and 10 min after reperfusion, respectively. Infarct size ( IS) and area at risk ( AAR) were deter-mined by 2, 3, 5-triphenyl-tetrazolium (TTC) staining at the end of reperfusion. Left ventricular developed pressure ( LVDP), ± dp/dtmax and heart rate ( HR) were recorded to evaluate cardiac function in each group. Results When compared with IR group, RPC significantly reduced IS / AAR and decreased the ac-tivity of LDH at 5 min and 10 min after reperfusion. However, SP600125 almost thoroughly abolished the protective effects of RPC, as evidenced by the in-creased value of IS / AAR and the high activity of LDH. In addition, PD98059 also partly blocked the effects of RPC, while SB203580 showed no influence on RPC. Meanwhile, the hemodynamic parameters such as LVDP, HR and ± dp/dtmax were not signifi-cantly different in any group except sham group. Con-clusion JNK and ERK pathways may play an impor-tant role in cardioprotective effects of remifentanil pre-conditioning against ischemia/reperfusion injury in rats with heart failure.
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Objective To analyze the computed tomography(CT)and magnetic resonance imaging (MRI)appearances of primary clear cell carcinoma of the liver (PCCCL)and evaluate the value in the diagnosis of the disease.Methods CT and MR images of 38 patients of pathologically confirmed PCCCL were evaluated retrospectively.Twenty-six patients underwent CT,23 underwent MRI, and 1 1 underwent both CT and MRI.Results In pre-contrast CT scanning,24 PCCCLs appeared hypodense and 2 hyperdense.As for MRI 1 9 of the 23 PCCCLs were hypointense and 4 were iso-hypointense on T1 WI.While on T2 WI,22 cases were heterogeneously hyperintense,and 1was iso-hypointense.On the arterial phase of CT/MRI,all cases presented intense enhancement,and on the portal venous phase,35 cases(35/38,92.1%)were relatively hypodense/hypointense and 3 were slightly hyperdense/hyperintense.Among the tumors larger than 3 cm(n=22),nodular enhancement pattern was found in 14 cases(63.6%,14/22).The capsular rim en-hancement was demonstrated in 26 cases.Conclusion PCCCL showed similar dynamic enhancement pattern as common hepatocellu-lar carcinoma,but also depicted specific imaging features.
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Objective To evaluate the roles of 1-phosphatidylinositol 3-kinase (PI3K) and extracellular signal-regulated kinase (ERK) signaling pathways in reduction of ischemia-reperfusion (I/R) injury to the isolated hearts by morphine preconditioning in the rats with chronic heart failure.Methods Adult male Sprague-Dawley rats,weighing 200-230 g,in which doxorubicin 2.0 mg/kg was injected via the tail vein once a week for 6 weeks to induce chronic heart failure,were studied.At the end of 8th week,42 rats with chronic heart failure were randomly divided into 7 groups (n =6 each) using a random number table:sham operation group (group S),I/R group,morphine preconditioning group (group MP),PD98059 (ERK inhibitor) + morphine preconditioning group (group PD + MP),wortmannin (PI3K inhibitor) + morphine preconditioning group (group WT + MP),PD98059 group (group PD) and wortmannin group (group WT).The hearts were quickly excised and passively perfused in a Langendorff apparatus and subjected to 30 min of occlusion of the left coronary artery followed by 2 h of reperfusion to establish the model of I/R injury.In group S,the hearts were only sutured,but not ligated and were continuously perfused with K-H solution for 195 min.In group I/R,the hearts were perfused with K-H solution for 45 min before ischemia.In group MP,the hearts were perfused with K-H solution for 15 min,with K-H solution containing morphine 1 μmol/L for 5 min and then with K-H solution for 5 min (3 cycles in total) before ischemia.In PD + MP and WT + MP groups,the hearts were perfused with K-H solution containing PD98059 (10 μmol/L) and wortmannin (100 nmol/L),respectively,starting from 10 min before morphine preconditioning until 5 min of ischemia.In PD and WT groups,the hearts were perfused with K-H solution containing PD98059 (10 μmol/L) and wortmannin (100 nmol/L),respectively,starting from 40 min before ischemia until 5 rin of ischemia.At 15 min of equilibration (baseline) and 5 and 10 min of reperfusion,the coronary flow was collected to detect the activity of lactate dehydrogenase (LDH).Infarct size (IS) and area at risk (AAR) were measured at the end of reperfusion and IS/AAR ratio was calculated.Results Compared with group S,LDH activity was significantly increased at 5 and 10 min of reperfusion,IS and IS/AAR ratio were also increased (P ＜ 0.05),and no significant change was found in AAR in group I/R (P ＞ 0.05).Compared with group I/R,LDH activity was significantly decreased at 5 min of reperfusion,IS and IS/AAR ratio were also decreased (P ＜ 0.05),and no significant change was found in AAR in group MP,and no significant change was found in LDH activity,IS,AAR and IS/AAR ratio in WT and PD groups (P ＞ 0.05).Compared with group MP,LDH activity was significantly increased at 5 and 10 min of reperfusion (P ＜ 0.05),and IS and IS/AAR ratio were decreased in group PD + MP,and no significant change was found in LDH activity,IS,AAR and IS/AAR ratio in group WT + MP (P ＞ 0.05).Conclusion Activation of ERK signaling pathway is involved in reduction of I/R injury to isolated hearts by morphine preconditioning in rats with chronic heart failure,however,PI3K signaling pathway has no such effect.
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Objective To evaluate the effects of morphine preconditioning on the expression of microRNAs (miRNAs) during hypoxia-reoxygenation (H/R) in isolated cardiomyocytes in rats with heart failure.Methods Healthy adult male Sprague Dawley rats,w eighing 200-220 g,were used in this study.Adriamycin 2.0 mg/kg was injected once a week for 6 weeks via the tail vein to induce heart failure.The cardiomyocytes were isolated from the failing hearts of rats and seeded in 24-well plates or in 60 mm diameter dishes.The cells were then randomly divided into 3 groups (n =16 each) using a random number table:control group (group C); group H/R;morphine preconditioning group (group MP).The cells were cultured in normal culture atmosphere in group C.After being exposed to hypoxic air (5％ CO2-95％ N2) for 90 min,the cells were returned to the high-glucose DMEM supplemented with 10％ newborn bovine serum and were then cultured for 120 min in H/R and MP groups.In group M,the cells were cultured in morphine culture medium (final concentration of morphine 0.3 μmol/L) for 10 min and then were returned to the culture medium without morphine and cultured for 30 min immediately before hypoxia.At 120 min of reoxygenation,the cells of 8 wells in each group were chosen to detect the cell viability and lactate dehydrogenase (LDH) activity (by Typan blue staining).All the RNAs were extracted from the cardiomyocytes of the left 8 wells in each group and subjected to miRNA microarray to screen differentially expressed miRNAs.Results The cell viability was significantly lower,the activity of LDH was higher,the expression of miR-6216 and let7e-5p was higher,and the expression of miR-133b-5p was lower in H/R and MP groups than in group C (P ＜ 0.05).Compared with H/R group,the cell viability was significantly increased,the activity of LDH was decreased,the expression of miR-133b-5p was up-regulated,and the expression of miR-6216 and let-7e-5p was down-regulated in MP group (P ＜ 0.05).Conclusion Morphine preconditioning reduces H/R injury to isolated cardiomyocytes in rats with heart failure through regulating the expression of miRNAs such as miR133b-5p,miR-6216 and let-7e-5p.
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<p><b>OBJECTIVE</b>To prepare the antibodies against salbutamol (SAL) with high sensitivity and to develop an indirect competitive enzyme-linked immunoassay (ic-ELISA) for fast detection of SAL.</p><p><b>METHODS</b>The New Zealand white rabbits were immunized with SAL in a small dose and long period mode. The method of ic-ELISA was optimized and adopted for the detection of a series of SAL samples, then the standard curve of SAL was established. The precision and the recoveries of the method were determined.</p><p><b>RESULTS</b>The antibodies with high sensitivity towards SAL were prepared with a IC50 of 12.21 ng/ml. The ic-ELISA method for SAL measurement was established, the recoveries of measurement was between 95%-105% and the CV was <3%.</p><p><b>CONCLUSION</b>The antibodies against salbutamol have been prepared and an indirect competitive enzyme-linked immunoassay for fast and specific detection of SAL has been developed.</p>
Subject(s)
Animals , Male , Rabbits , Albuterol , Allergy and Immunology , Antibodies , Allergy and Immunology , Enzyme-Linked Immunosorbent Assay , MethodsABSTRACT
<p><b>OBJECTIVE</b>To synthesize artificial diethylstilbestrol (DES) antigen and to prepare DES polyclonal antibody with high titer and sensitivity.</p><p><b>METHODS</b>The derivative of DES (DES-HS) was synthesized from diethylstilbestrol, ethyl bromoacetate,bovine serum albumin (BSA) and chicken ovalbumin (OVA) with the nucleophilic substitution reaction; the compound was identified by electrospray ionization mass spectrometry(ESI-MS). The DES-HS and the carrier proteins (BSA, OVA) were cross-linked to prepare the artificial antigen; the UV absorption spectrophotometry and high performance liquid chromatography (HPLC) were used to identify the prepared artificial antigen. The rabbits were immunized with the DES artificial antigen to prepare the DES polyclonal antibodies.</p><p><b>RESULTS</b>The DES-HS was synthesized. The DES artificial antigen was prepared successfully with a coupling rate of 22:1. The DES polyclonal antibodies with a titer of 1:25 600 and IC50 of 10.81 ng/ml were prepared with DES artificial antigen.</p><p><b>CONCLUSION</b>A set of methods to synthesize DES artificial antigen and to prepare the DES polyclonal antibodies has been developed successfully.</p>
Subject(s)
Animals , Male , Rabbits , Antibodies , Allergy and Immunology , Antigens , Chemistry , Allergy and Immunology , Diethylstilbestrol , Allergy and Immunology , Enzyme-Linked Immunosorbent Assay , MethodsABSTRACT
Objectlve To evaluate the clinical value of the detection of Myeobacterium tuberculosis (MTB)and its L-forms(MTB-L)in pefipheral blood.Methods MTB and MTB-L in blood samples of 156 patients with tuberculosis and 147 patients with lung cancer were detected by hemolysis centrifugal drop Intensified Kinyoun's acid fast staining (IK) and hemolysis centrifugal culture with 42 healthy persons as controls.MTB DNA was detected by TaqMan-PCR technique in plasma mononuclear cells and whole blood from aforementioned groups.Results Among each groups detected by IK.the MTB positive rate was 1.3%(2/156),0.7%(1/147),and 0%(0/42)respectively.The positive rate of culture was 0.6%(1/156),0%(0/147),and 0%(0/42)respectively.MTB-L positive rate detected by IK was 41.0%(64/156),32.7%(48/147)and 7.2%(3/42).MTB-L positive rate detected by culture was 25.6%(40/156)and 39.5%(58/147).0%(0/42).MTB positive rate were significantly different from the MTB-L positive rate (P＜0.01).Among the 98 strains L-forms,81 strains were from human,17 strains were from bovis,and 44 strains were back to the ancestors.The positive rate of mononuclear ceHs and whole blood obtained from the tuberculosis group detected by TaqMan-PCR were 77.6%(121/156) and 68.6%(107/156)(P＜0.05).The positive rate of mononuclear cells and whole blood from lung cancer group were 59.2%(87/147)and 48.3%(52/147)(P＜0.05),which were higher than the plasma positive rate 12.2%(18/147)(P＜0.05).The result of individual bloed constituents (except for plasma) from two groups were significantly different from the control(P＜0.01).Conclusions MTB-L was found in the peripheral blood of the tuberculosis and lung cancer patients.Hemolysis centrifugal culture is a rapid,simple and reliable method for detecting MTB-L TaqMan-PCR showed the positive rate of MTB DNA in plasma and whole blood samples were significantly higher than that in the plasma.
ABSTRACT
Objective:To explore the apoptosis inducing effect of matrine on C6 glioma cells and the related mechanisms.Methods:MTT assay was used to examine the inhibition of C6 glioma cell line by matrine at various concentrations and the IC50 was calculated.Inverted microscope and TEM(transmission electron microscope) were employed to observe the morphological alterations of C6 glioma cells after exposure to matrine;FCM(Flow cytometry) was used to detect the apoptosis rate of C6 glioma cells;and real-time PCR was used to examine the differential expression of related genes.ICC and Western blotting was used to detect the expression of caspase-3.Results:MTT showed that the cell inhibition effect of matrine increased with its concentration(0.1-1.0 mg/ml)(P