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Objective: To investigate the clinicopathological features, immunophenotype, diagnosis and differential diagnosis of SRF-rearranged cellular perivascular myoid tumor. Methods: Two cases of SRF-rearranged cellular perivascular myoid tumor diagnosed in the Department of Pathology, Fudan University Shanghai Cancer Center from October 2021 to March 2022 were collected. Immunohistochemical staining, fluorescence in-situ hybridization (FISH) and next-generation sequencing (NGS) were performed, and the literature was reviewed. Results: Case 1, a 3-month-old boy presented with a painless tumor of the scalp, measuring about 2 cm in diameter. Case 2, a 3-year-old girl complained with a painless tumor of the knee, measuring approximately 1.5 cm in diameter. Microscopically, the tumor had a clear boundary and showed multinodular growth. The tumor was mainly composed of spindle cells arranged in long intersecting fascicles associated with thin, slit-like or branching ectatic vessels, focally forming hemangiopericytoma-like appearance. The tumor cells were abundant, but there was no obvious atypia. Mitotic figures (3-4/10 HPF) were noted. H-caldesmon and SMA were positive in both cases. Case 1 showed diffuse and strong positivity for Desmin, and focally for CKpan. Ki-67 proliferation index was 20% and 30%, respectively. FISH displayed NCOA2 gene translocation in case 1 and the RELA gene translocation in case 2. NGS detected the SRF-NCOA2 gene fusion in case 1 and the SRF-RELA gene fusion in case 2. Both patients underwent local excisions. During the follow-up of 5-14 months, case 1 had no local recurrence, while case 2 developed local recurrence 1 year post operatively. Conclusions: SRF-rearranged cellular perivascular myoid tumor is a novel variant of perivascular cell tumor, which tends to occur in children and adolescents. The tumor forms a broad morphologic spectrum ranging from a pericytic pattern to a myoid pattern, and include hybrid tumors with a mixture of pericytic and myoid patterns. Due to its diffuse hypercellularity and increased mitotic figures and smooth muscle-like immunophenotype, the tumor is easy to be misdiagnosed as myogenic sarcomas. The tumor usually pursues a benign clinical course and rare cases may locally recur.
الموضوعات
Child, Preschool , Female , Humans , Infant , Male , Biomarkers, Tumor/analysis , Calmodulin-Binding Proteins , China , Hemangiopericytoma/pathology , Sarcoma/pathology , Soft Tissue Neoplasms/pathologyالملخص
【Objective】 To identify the specificity of alloantibody against high-frequency antigens in one case suffering with severe hemolytic diseases of the fetus and newborn (HDFN) and to screen for matching blood for transfusion. 【Methods】 The HDFN test and the antibody serological identification tests in the mother were performed. Several common high frequency antigens of maternal red blood cells (RBCs) were determined. IgG subtype coated on the RBCs of the newborn was determined. The phagocytic efficiency of the antibody was tested using the monocyte phagocytosis of sensitized erythrocyte by flow cytometry in vitro. Sanger sequencing of DI gene was performed in the mother, father and mother’s brother. The diluted maternal plasma was used for large scale screening of matching blood using IAT in Coomb’s gel card. 【Results】 Di(b-) phenotype was identified in the mother of the newborn and anti-Dib (titer: 512) related HDN was detected in the newborn. IgG1 and IgG2 subtypes of anti-Dib were detected and the rate of monocyte phagocytosis was 88.83%(74.7/84.09). The compatible blood was not detected in the maternal relatives. Subsequently, the newborn received the matching RBCs of two Di(b-) donors identified from 5 520 blood donors and discharged from the hospital. We screened out 17 Di(b-) donors out of 51 334 blood donors, indicating that the distribution frequency of Di(b-) among blood donors in Guangzhou was about 0.033% (17/51 334). 【Conclusion】 By serology and molecular biology methods, the newborn was identified with HDFN caused by anti-Dib, and an effective large-scale screening method for Di (b -) rare blood types was established to find matching blood, which supported the establishment of rare Di(b-) blood database.
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Retzius-sparing robot assisted radical prostatectomy (RS-RARP) can significantly improve the immediate urinary continence without increasing the positive rate of surgical margin.However, the learning curve is long, and fewer than 10% of the surgeons can master it.Therefore,we have optimized the procedures of RS-RARP, applying radical prostatectomy with retrograde release of neurovascular bundle to preserve it to the maximum extent.Urethral anastomosis can be performed with only one suture, which eliminates the need for Hem-o-lok and reduces subsequent complications.Our team routinely carries out this operation, and conlcudes that this surgical method can achieve good tumor control, good urinary continence, fast recovery of sexual function, few complications, and strong operability.This article details the key steps and operation experience of this technique.
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【Objective】 To elucidate the prediction ability of monocyte monolayer assay(MMA) used in hemolytic disease of fetus and newborn(HDFN) caused by IgG anti-M. 【Methods】 Plasma from eight pregnant women containing IgG anti-M were collected, and were divided into two groups(4 cases with HDFN, with severe clinical symptoms such as fetal hydrops, and 4 cases without HDFN) according to the clinical outcomes. M antigen positive cells were sensitized with dithiothreitol(DTT) treated plasma from eight pregnant women respectively. MMA was performed by coincubation with monocytes and sensitized M cells, along with negative and positive control set up. T-test was conducted to compare the difference in phagocytic efficiency between two groups. 【Results】 The phagocytic efficiency in group with HDFN were 15.37%, 13.05%, 9.17% and 24.50% respectively, with the mean value of 15.52%, while the group without HDFN were 8.74%, 11.07%, 5.12% and 6.23% respectively, with the mean value of 7.79%.There was no significant difference in phagocytic efficiency between two groups(P>0.05). The mean values of both groups were not significantly different from the negative control(P>0.05), but both were significantly lower than positive control(P<0.05). 【Conclusion】 The low phagocytic efficiency couldn’t convince that the MMA is an effective predictor for the HDFN caused by IgG anti-M, indicating that another mechanism might be responsible for it rather than monocyte phagocytosis. The assessment of the peak systolic velocity in middle cerebral artery of the fetal should be considered in the management for pregnant women who produce IgG anti-M to estimate the situation of fetal anemia.
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Objective:To screen key genes of renal clear cell carcinoma based on bioinformatics methods, identify possible microRNA (miRNA)-mRNA action axis, and explore the expression of related genes in clear cell renal cell carcinoma tissues and cells.Methods:Gene expression profiles of GSE40435 and GSE71302 datasets were obtained from the Gene Expression Omnibus (GEO) database. TCGA-KIRC datasets were obtained from The Cancer Genome Atlas (TCGA) database. R software was used to identify the differentially expressed mRNA and miRNA, and the functional enrichment analysis was performed. STRING database and Cytoscape software were used to perform the protein interaction analysis. The prognosis-related differentially expressed miRNA was evaluated by the Oncomir database. The potential targeted genes regulated by miRNA were determined by using TargetScan and miRDB targeted gene prediction tools. The tissue samples and clinicopathological features of 34 patients with clear cell renal cell carcinoma in the First Hospital of Shanxi Medical University from June to December 2021 were collected, and normal renal cell line 293T and clear cell renal cell carcinoma cell line 786O were selected. The real-time fluorescence quantitative polymerase chain reaction (qRT-PCR), was used to detect the relative expression of genes; Western blotting and immunohistochemical staining were used to detect the expression levels of the targeted proteins. The dual luciferase reporter gene assay was carried out to verify the targeting relationship between genes.Results:A total of 1 351 differentially expressed mRNA and 50 differentially expressed miRNA were screened and identified. The result of functional enrichment analysis suggested that the fatty acid metabolism pathway and xenobiotic metabolism pathway were suppressed in clear cell renal cell carcinoma, while the apoptosis and immune response pathways were activated. Protein interaction analysis suggested that the signal transduction and protein ubiquitination pathways might play a key role in clear cell renal cell carcinoma. The screening results showed that miRNA-224-5p (miR-224-5p) was most closely associated with clear cell renal cell carcinoma progression and was highly expressed in tumor tissues, and its prognosis-related target gene was NEDD4L. The relative expression of NEDD4L mRNA in clear cell renal cell carcinoma tissues and paraneoplastic tissues were 0.138±0.103 and 1.000±0.026 ( t = 46.23, P < 0.05), and the relative expression of miR-224-5p was 1.000±0.043 and 0.129±0.108 ( t = 45.28, P < 0.05). The differences of NEDD4L mRNA and miR-224-5p expressions in different grades and stages of clear cell renal cell carcinoma tissues were statistically significant (all P < 0.05). The expression of NEDD4L protein was decreased in clear cell renal cell carcinoma. The relative expression of NEDD4L gene in 293T and 786O cells were 1.000±0.125 and 0.210±0.044 ( t = 17.52, P < 0.05); the relative expressions of miR-224-5p gene were 0.209±0.049 and 1.000±0.234 ( t = 10.61, P < 0.05). The relative expressions of NEDD4L mRNA in miRNA mimic group and negative control group were 0.236±0.062 and 1.000±0.024, and the difference was statistically significant ( t = 43.56, P < 0.05). NEDD4L protein expression was reduced in the miRNA mimic group. Dual luciferase reporter gene assay suggested that NEDD4L was a direct target gene of miR-224-5p. Conclusions:In clear cell renal cell carcinoma, miR-224-5p targets and regulates NEDD4L expression, and this mechanism may be related to carcinogenesis and progression of clear cell renal cell carcinoma.
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【Objective】 To solve the difficulty of RhD blood group typing in a patient with double population(DP) of red blood cells for RhD antigen by serological and genotyping analysis. 【Methods】 Separation of the two populations of red blood cells of the patient was performed using capillary centrifugation method. ABO, RhD and RhCE typing, direct anti-human globulin test (DAT), irregular antibody screening, antibody identification and blood crossmatching of the patient were conducted using the standard serological methods. The hybrid Rhesus zygosity analysis of the RHD gene was performed by PCR-RFLP method. RHD and RHCE genotype of the patients were identified by PCR-SSP method. 【Results】 The patient was B type but with DP of red blood cells for RhD, Rhc and RhE antigens. DAT of the patient was positive and the alloanti-D was detected in serum. The RHD zygosity was D-/D- homozygote. PCR-SSP testing showed the RHD gene deletion (RHD * 01N. 01/01N.01 genotype) and Ccee of RHCE genotype in the patient, which was consistent with RHD zygosity analysis. 【Conclusion】 This is a special case with D-negative phenotype which was wrongly detected as D-positive type after D-positive red blood cells transfusion in emergency. When the DP of red cells for D antigen encountered like this case, the RhD typing can be accurately determined by using RHD genotyping analysis to provide strong evidence to the clinical blood transfusion.
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Antibodies against human CD36 are responsible for several immune-mediated disorders. The detection of anti-CD36 antibodies using the standard monoclonal antibody (mAb) immobilization of platelet antigens (MAIPA) assay is hampered by a high frequency of false-negative results, most likely due to competitive inhibition of the mAb used as the capture antibody. We generated a panel of mouse mAbs against CD36 and seven hybridomas (GZ-3, GZ-13, GZ-70, GZ-143, GZ-413, GZ-507, and GZ-608), which were selected for MAIPA assays, as they reacted with mouse and human CD36. Fourteen anti-CD36 sera were assayed; all of which showed a positive reaction in a PakPlus (Immucor GTI Diagnostics, Inc., Waukesha, WI, USA) ELISA-based screening (optical density: 0.257–2.292). When the reference anti-CD36 mAb FA6-152 was used in the MAIPA assay, only 6/14 (42.9%) sera displayed a positive reaction. In contrast, anti-CD36 antibodies were detected in 13/14 (92.9%) sera when GZ-70 and GZ-608 mAbs were used. This significant improvement resulted in the identification of anti-CD36 antibodies by an antigen capture assay. Since patient’s platelets possibly carrying rare native antigens are used, this method will facilitate the identification of new platelet antibodies against CD36 that are involved in immune-mediated thrombocytopenia and other diseases, such as transfusion-related acute lung injury.
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BackgroundThe diagnosis of Alzheimer's disease （AD） still faces great challenges， and the advantage of electroencephalogram （EEG） diagnosis lies in its portable and non-invasive nature， so the EEG diagnosis of AD has occupied an important place in clinical research. ObjectiveTo evaluate the value of resting state EEG for AD diagnosis， and to provide references for early recognition of AD in clinical practice. MethodsClinical data of AD patients （n=59） in an Inpatient Geriatric Psychiatry Unit of Shenzhen Kangning Hospital from May 2019 to May 2022 were retrospectively analyzed， and healthy elderly individuals attending outpatient clinics at the hospital during the same period were enrolled as control group （n=54）. Eight-channel resting state EEG data were acquired， and the absolute power values in the α， β， θ and δ frequency bands and the α/θ ratio were obtained and calculated using Fast Fourier Transform （FFT）. Cognitive function assessments of patients were done by Mini-Mental State Examination （MMSE） and Montreal Cognitive Assessment （MoCA）. Spearman correlation analysis was used to examine the correlation between EEG findings and MMSE and MoCA scores of AD patienrs. Logistic regression prediction model for AD was built using currently available EEG and clinical variables， and the model performance was assessed using the receiver operating characteristic （ROC） curve and the area under curve （AUC）. ResultsThe θ-band absolute powers in the right mid-frontal （F4） and mid-lateral （F7， F8） regions were higher in AD patients than those in healthy controls， with statistically significant difference （t=-2.844， -2.825， -3.014， P<0.05 or 0.01）. The absolute powers of α/θ ratio in prefrontal （Fp1， Fp2）， mid-frontal （F3， F4） and mid-lateral （F7， F8） regions showed a notable reduction in AD patients compared with healthy controls， with statistical difference （t=2.081， 2.327， 3.423， 2.358， 3.272， 2.445， P<0.05 or 0.01）. Spearman correlation analysis denoted that MMSE score was positively correlated with the absolute powers of α-band， β-band and α/θ ratio （r=0.206， 0.288， 0.372， P<0.05 or 0.01）. MoCA score was positively correlated with β absolute powers and α/θ ratio （r=0.201， 0.315， P<0.05 or 0.01）， and negatively correlated with θ absolute power （r=-0.218， P<0.05）. ROC curve revealed an AUC of 0.882 （95% CI： 0.820~0.943）， a sensitivity of 0.966 and a specificity of 0.673 for the AD prediction model based on EEG variables， while the prediction model for AD using comprehensive variables achieved better predictive efficacy， reaching an AUC， sensitivity and specificity of 0.946 （95% CI： 0.905~0.986）， 0.948 and 0.873， respectively. ConclusionResting state EEG of AD patients is correlated with cognitive function， and are of great value in the diagnosis of AD， with θ absolute power and α/θ ratio in EEG being the most strongly correlated with AD.
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Two new species of Fulvifomes are described from specimens collected in rainforests of Nonggang Nature Reserve of southern China, based on morphological characteristics and molecular phylogenetic analysis of the internal transcribed spacer (ITS) and nuclear large subunit ribosomal DNA (nLSU) sequences. Fulvifomes nonggangensis sp. nov. is characterized by perennial, sessile and solitary basidiocarps, applanate pileus, small cystidioles of 9.9–15.4 Â 2.9–3.5 lm, large pores of 5–6 per mm, a dimitic hyphal system, and broadly ellipsoid basidiospores of 4.3–5.3 Â 3.3–4.2 lm. F. tubogeneratus sp. nov. is characterized by perennial, sessile, and imbricate basidiocarps, a duplex context, small pores of 7–8 per mm, a dimitic hyphal system, and ovoid to subglobose basidiospores of 5.72 Â 5.00 lm.
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Two new species of Fulvifomes are described from specimens collected in rainforests of Nonggang Nature Reserve of southern China, based on morphological characteristics and molecular phylogenetic analysis of the internal transcribed spacer (ITS) and nuclear large subunit ribosomal DNA (nLSU) sequences. Fulvifomes nonggangensis sp. nov. is characterized by perennial, sessile and solitary basidiocarps, applanate pileus, small cystidioles of 9.9–15.4 Â 2.9–3.5 lm, large pores of 5–6 per mm, a dimitic hyphal system, and broadly ellipsoid basidiospores of 4.3–5.3 Â 3.3–4.2 lm. F. tubogeneratus sp. nov. is characterized by perennial, sessile, and imbricate basidiocarps, a duplex context, small pores of 7–8 per mm, a dimitic hyphal system, and ovoid to subglobose basidiospores of 5.72 Â 5.00 lm.
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Seven new triterpenoid saponins, including five ursane-type saponins, ilexchinenosides R-V (1-5), and two oleanane-type saponins, ilexchinenosides W-X (6-7), with four known triterpenoid saponins (8-11) were isolated from the leaves of Ilex chinensis. Their structures were elucidated by comprehensive spectroscopic 1D and 2D NMR and HR-ESI-MS data. Their sugar moieties were determined by HPLC analysis compared with standards after hydrolysis and derivatization. Compounds 1, 2, 4, 9 and 10 exhibited potential hepatoprotective activity against N-acetyl-p-aminophenol (APAP)-induced HepG2 cell injury in vitro.
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【Objective】 To find the HLA-matched platelets from platelet donor registry and track the transfusion effect for aplastic anemia patients in pregnancy with platelet transfusion refractory (PTR) caused by anti-HLA, so as to support the childbirth and follow-up treatment of the patients. 【Methods】 Antibodies to HPA and HLA were detected by ELISA kit and Luminex, respectively. DNA of the patient and 523 platelet donors from the donor registry were extracted for high-resolution HLA genotyping. The Risk Factors Evaluation Software of PTR was used to select the ABO-compatible and HLA-matched donors, without HLA Eplets specific to the patient. After MASPAT cross matching, the patient was transfused with 1 U of platelets, and the 24h post-transfusion effect was recorded. 【Results】 Only anti-HLAⅠantibody was found in the patient serum, and the specificity Eplet was 65QIA, including HLA-B*27∶08, B*27∶05, B*07∶02, B*55∶01, B*67∶01 and B*54∶01; anti-HLA Ⅱ antibody was negative. The HLA genotypes of both the patient and donor were HLA-A*02∶07, A*11∶01, B*46∶01, B*46∶01, C*01∶02, 01∶03, DRB1*04∶05, DRB1*0901, DQB1*03∶03 and DQB1*04∶01. The results of MASPAT matching were negative. HLA-matched platelets transfusion provided a satisfactory posttransfusion platelet responses in patients(1 before vs 33 ×109/L after). A baby boy was delivered by cesarean section 4 weeks later, and the same donor was recruited due to the mother′s low Plt and bleeding trend. The 24h posttransfusion Plt (×109 / L) rose from 5 to 37 after the secondary transfusion of 1U platelet. The vital signs of the mother and her baby were normal during the two-day follow up. 【Conclusion】 The establishment of blood donor registry and screening of HLA-matched donors is an effective approach to treat PTR caused by HLA antibodies in pregnancy complicated with aplastic anemia.
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BACKGROUND@#The optimal treatment for large impacted proximal ureteral stones remains controversial. The aim of this study was to evaluate the efficacy, safety, and potential complications of mini-percutaneous nephrolithotomy (MPCNL) and retroperitoneal laparoscopic ureterolithotomy (RPLU) in the treatment of impacted proximal ureteral stones with size greater than 15 mm.@*METHODS@#A total of 268 patients with impacted proximal ureteral stones greater than 15 mm who received MPCNL or RPLU procedures were enrolled consecutively between January 2014 and January 2019. Data on surgical outcomes and complications were collected and analyzed.@*RESULTS@#Demographic and ureteral stone characteristics found between these two groups were not significantly different. The surgical success rate (139/142, 97.9% vs. 121/126, 96.0%, P = 0.595) and stone-free rate after 1 month (139/142, 97.9% vs. 119/126, 94.4%, P = 0.245) of RPLU group were marginally higher than that of the MPCNL group, but there was no significant difference. There was no significant difference in the drop of hemoglobin between the two groups (0.8 ± 0.6 vs. 0.4 ± 0. 2 g/dL, P = 0.621). The mean operative time (68.2 ± 12.5 vs. 87.2 ± 16.8 min, P = 0.041), post-operative analgesics usage (2/121, 1.7% vs. 13/139, 9.4%, P = 0.017), length of hospital stay after surgery (2.2 ± 0.6 vs. 4.8 ± 0.9 days, P < 0.001), double J stent time (3.2 ± 0.5 vs. 3.9 ± 0.8 days, P = 0.027), time of catheterization (1.1 ± 0.3 vs. 3.5 ± 0.5 days, P < 0.001), and time of drainage tube (2.3 ± 0.3 vs. 4.6 ± 0.6 days, P < 0.001) of MPCNL group were significantly shorter than that of the RPLU group. The complication rate was similar between the two groups (20/121, 16.5% vs. 31/139, 22.3%, P = 0.242).@*CONCLUSIONS@#MPCNL and RPLU have similar surgical success and stone clearance in treating impacted proximal ureteral stones greater than 15 mm, while patients undergoing MPCNL had a lower post-operative pain rate and a faster recovery.
الموضوعات
Humans , Laparoscopy , Length of Stay , Nephrolithotomy, Percutaneous/adverse effects , Retroperitoneal Space/surgery , Treatment Outcome , Ureteral Calculi/surgeryالملخص
Objective:To establish ultra-high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） for the analysis of 18 components of five categories, namely spirosteroid saponins （timosaponin AⅠ，timosaponin AⅡ，timosaponin AⅢ，anemarrhenasaponin Ⅲ，sarsasapogenin），furostane saponins（anemarrhenasaponin Ⅰ，anemarrhenasaponin Ⅰa，anemarsaponin E，officinalisinin I，timosaponin B-Ⅱ，timosaponin B-Ⅲ，anemarsaponin C），flavonoids（icarisin I，baohuoside I），bisphenone（meomangiferin，mangiferin，isomangiferin），and hydroquinone glycoside （β-arbutin）,in order to analyze the differences between the main root and fibrous root of Anemarrhena asphodeloides from different sources and provide reference for the sustainable development of A. asphodeloides resources. Method:0.25 g sample was refluxed and extracted with 25 mL dilute ethanol for 30 min. The chromatographic separation was carried out on Waters Acquity Uplc BEHHILI C18 column（2.1 mm×100 mm，1.7 μm），with 0.1% formic acid water-acetonitrile as the mobile phase for gradient elution，and the volume flow rate was 0.2 mL·min-1. Electrospray ion source（ESI+，ESI-），and multi-reaction ion monitoring（MRM） were used for detection，external standard method was used to calculate the content of the tested components in medicinal samples，and SMICA14.1 software was used to analyze the differences between the main root and fibrous root samples of A. asphodeloides. Result:The tested components showed a good linear relationship in their respective linear ranges，with a good precision，repeatability and stability. The recovery rate of samples was between 95.22%-101.42%，and RSD was less than 4%. The experimental results showed great differences between the main root and fibrous root of A. asphodeloides, when the multivariate statistical analysis was carried out with 18 main components. Conclusion:This study provides experimental basis for the reuse of fibrous root of A. asphodeloides resources and the quality control of A. asphodeloides.
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This study is to improve the quality standard and supply the scientific basis for Anemarrhenae Rhizoma and its raw processed products. Steroidal saponin including timosaponin BⅡ, timosaponin AⅢ and flavonoids including neomangiferin and mangiferin were selected as the indicative components. Silica gel G thin layer chromatography(TLC) and polyamide TLC were used to detect the two types of compounds, respectively. The contents of timosaponin BⅡ and timosaponin AⅢ were determined by HPLC-ELSD and the content of neomangiferin, mangiferin and isomangiferin were determined by HPLC-UV. Moisture, total ash and acid insoluble ash were determined according to Chinese Pharmacopoeia(2015 edition). And 80% ethanol was selected as the solvent and the content determination of total extract were determined. The fingerprints of Anemarrhenae Rhizoma and its raw processed products were established by HPLC-UV and HPLC-ELSD. The results showed that the methods of TLC and HPLC have been successfully stablished. There are 2 and 3 peaks which have been identified by HPLC-ELSD and HPLC-UV, respectively. The HPLC fingerprint methods are specific and can be used to identify and quality control for Anemarrhenae Rhizoma and its raw processed products in the mass. Comparing to Chinese Pharmacopoeia(2015 edition), the TLC identification and content determination were revised and the total extract determination and HPLC fingerprints were added in the present study. Our results can be used as the scientific basis of quqlity control for Anemarrhenae Rhizoma and its raw processed products.
الموضوعات
Anemarrhena , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Reference Standards , Rhizomeالملخص
OBJECTIVE@#To construct eukaryotic expression vectors for human platelet CD36 gene 220 C>T and 429+4insg variants and analyze their expressions in HEK293T cells.@*METHODS@#RNA was isolated from human platelets and reversely transcribed into cDNA. Sequences of 220C>T and 429+4insg variants were derived by PCR amplification. The target sequence was ligated into a pcDNA3.1/V5-His-TOPO vector by TA cloning, which was transformed into TOP10 E. coli. Positive plasmids were screened by blue-white selection. After sequencing, plasmid DNA carrying 220C>T or 429+4insg variant was used to transfect HEK293T cells with the help of effectene. Expression of CD36 protein was then analyzed by flow cytometry and Western blotting.@*RESULTS@#An eukaryotic expression vector pcDNA3.1/V5-His-CD36 (220C>T/429+4insg) was constructed by TA cloning. After transfected into HEK293T cells, the 220C>T and 429+4insg variants resulted in CD36 deficiency in HEK cells, which was confirmed by flow cytometry and Western blotting.@*CONCLUSION@#The 220C>T and 429+4insg variants can cause CD36 deficiency in human platelets. This system may be used for assessing the effect of 220C>T, 429+4insg, and other variants on the expression of CD36.
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Humans , Blood Platelets , CD36 Antigens , Cloning, Molecular , Escherichia coli , Eukaryota , Genetic Vectors , HEK293 Cells , Plasmids , Transfectionالملخص
Objective To explore the clinical feasibility of predicting synchronous liver metastases based on MRI radiomics nomogram based on T2WI in rectal cancer. Methods The imaging and clinical data of 261 patients with primary rectal cancer admitted to Zhejiang People′s Hospital from April 2012 to May 2018 were retrospectively analyzed. 101 patients were accompanied by synchronous liver metastasis All cases were divided into training group (n=182) and verification group (n=79). T2WI image of each patient was selected to extract texture features by AK analysis software of GE company. A radiomics signature was constructed after reduction of dimension in training group by the least absolute shrinkage and selection operator (LASSO). Univariate logistic regression was used to select for independent clinical risk factors and multivariate logistic regression along with imaging omics tags were used to construct predictive models and nomogram. ROC was used to assess the accuracy of the nomogram in the training group and to verify them by the validation group. Finally, the clinical efficacy of each patient′s synchronized liver metastasis risk factor was calculated based on the nomogram. Results A total of 328 texture features were extracted from the T2WI. Seven most valuable features were selected after reducing the dimension by LASSO algorithm, including 3 co-occurrence matrices (GLCM) and 4 run-length matrices(RLM). Tumor staging and radiomic signatures were included in the Multifactor logistic regression to build the prediction model and nomogram. The accuracy of predicting SRLM was 0.862 and 0.844 in the training and the verification group, respectively. To evaluate the accuracy of the nomogram, radiomics signature and the tumor staging in all cases were 0.857, 0.832 and 0.663, respectively. There was no significant difference in the number of SRLM cases between the high risk group and the low risk group based on nomogram (P>0.05). Conclusion The radiomics nomogram based on T2WI can be used as a quantitative tool to predict synchronous liver metastases of rectal cancer.
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In this work,a high performance liquid chromatography-ultraviolet( HPLC-UV) detection technology was used to establish fingerprint analysis method for Sanye Tangzhiqing Decoction following an analytical quality by design( AQb D) approach. Firstly,column temperature,flow rate,and gradient elution conditions were determined as the method parameters needing to be optimized. Then according to the results of definitive screening design,three critical method attributes( CMAs) were identified,including peak number,the percentage of common peak area to total peak area,and retention time of the last peak. A stepwise regression method was used then to build quantitative models between CMAs and method parameters. Probability-based design space was calculated and successfully verified using the experimental error simulation method. After the analysis conditions were optimized,the contents of six components,namely chlorogenic acid,paeoniflorin,rutin,hyperoside,quercetin-3-O-β-D-glucuronide,and salvianolic acid B were simultaneously determined. There were 19 common peaks in the fingerprint and their common peak area accounted for 96% of the total peak area. Both fingerprint and quantitative analysis methods were validated applicable in methodology study,and they can be applied to determine new samples.
الموضوعات
Chlorogenic Acid , Chromatography, High Pressure Liquid , Drugs, Chinese Herbalالملخص
Objective@#To explore the risk factors of predicting white matter hyperintensities progression based on radiomics of MRI of whole-brain white matter.@*Methods@#The imaging and clinical data of 152 patients with white matter hyperintensities admitted to Zhejiang People′s Hospital from March 2014 to October 2018 were retrospectively analyzed. The whole brain white matter on baseline T1WI images of each patient were segmented by SPM12 software package, and images of white matter were imported into AK software for texture feature extraction and dimensionality reduction. At last, least absolute shrinkage and selection operator(LASSO) was used to calculate the score of radiomics signature of each patient. According to the improved Fazekas scale, patients with WMH progression were divided into three groups: any white matter hyperintensities (AWMH), periventricular white matter hyperintensities (PWMH) and deep white matter hyperintensities (DWMH). Statistical differences of clinical factors and radiomics signature between WMH progression subgroups and non-progression subgroups were compared with independent sample t test or Mann-Whitney U test, and Univariate logistic regression was used to select independent clinical risk factors and multivariate logistic regression along with imaging omics tags were used to construct predictive models, which was evaluated by ROC curve. And the correlation between the clinical indicators of independent risk factors and the texture feature of radiomics signature was analyzed.@*Results@#The efficacy of the model for the detection for AWMH progression, PWMH progression and DWMH progression was 0.818,0.778 and 0.824, respectively. Age is an independent risk factor for AWMH progression and DWMH progression[OR(95%CI), 4.776(2.152-10.601) vs. 3.851(1.101-8.245); P<0.05]. BMI is an independent risk factor for DWMH[OR(95%CI), 3.004(1.204-7.370); P<0.05], and hyperlipidemia is an independent risk factor for AWMH and PWMH[OR(95%CI), 4.062(1.834-8.998) vs. 3.549(1.666-7.563); P<0.05]. In AWMH subgroup, Surface Area was negatively correlated with age and low density lipoprotein(LDL) (r=-0.401, -0.312), and Inverse Difference Moment_ALLDirection_offset 1_SD was negatively correlated with age(r=-0.412). In PWMH subgroup, Compactness 1 was negatively correlated with LDL(r=-0.198), and Inverse Difference Moment_angle 0_offset 7 was positively correlated with LDL(r=0.252). In DWMH subgroup, LongRunEmphasis_ALLDirection_offset 7 was negatively correlated with age(r=-0.322), and GLCMEntropy_angle0_offset 4 was negatively correlated with age(r=-0.278). GLCMEntropy_AllDirection_offset4 was negatively correlated with body mass index(r=-0.514).@*Conclusion@#Radiomics based on whole-brain white matter MR imaging can predict WMH progression and identify the risk factors in high-risk populations, thus providing early additional information to conventional magnetic resonance imaging to predict WMH progression.
الملخص
OBJECTIVE Glioblastoma multiforme (GBM) is the most malignant primary tumor of the central nervous system and is associated with a very poor prognosis. No further improvements in outcomes have been reported since radiotherapy-temozolomide therapy was introduced.Therefore,de-veloping new agents to treat GBM is important. This study aimed to evaluate the anti-tumor effect of evodiamine (Evo) on GBM cells, and to determine the underlying mechanisms involved. METHODS U251,LN229,HEB and PC12 cells were treated with various concentrations of evodiamine for 24 and 48 hours,cell viability was measured by MTT assay.The U251 and LN229 cells were treated with evo-diamine(0-10 μmol·L-1)for 24 h,and then stained with Hoechst 33258.An Annexin V-FITC Apoptosis Detection Kit was used to detect apoptosis in the cells.Reactive oxygen species(ROS)production was detected using dichlorofluorescein diacetate (DCFH-DA) staining. The changes in mitochondrial mem-brane potential (MMP) were assessed by JC-1 after cells were treated with evodiamine. The expres-sion levels of p-PI3K,PI3K,p-Akt,Akt,Bax,Bcl-2,p-p38,p38,p-JNK,JNK,p-ERK,ERK,Cytochrome c, Caspase-3, cleaved Caspase-3, PRAP, and cleaved PARP were measured by Western blot analy-ses. RESULTS According to MTT assay results, Evo significantly inhibited the cell proliferation in a time- and dose-dependent manner. Fluorescence microscopy and flow cytometry analyses revealed that Evo induced cell apoptosis in a concentration-dependent manner.Moreover,Evo induced reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) disruption. Finally, Evo induced apoptosis in cancer cells by suppressing PI3K/AKT signaling and inducing MAPK phos-phorylation(p38 and JNK,but not ERK)to regulate apoptotic proteins(Bax,Bcl-2,Cytochrome c,Cas-pase-3, and PARP). CONCLUSION In summary, Evo inhibits cell proliferation by inducing cellular apoptosis via suppressing PI3K/AKT and activating MAPK in GBM;these results indicate that Evo may be regarded as a new approach for GBM treatment.