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Liver biopsy is a key method for clarifying the diagnosis of liver diseases and has an important value in determining disease severity, deciding treatment timing, and predicting treatment response and prognosis. By recognizing the pattern of liver pathological injury, pathologists evaluate the nature of lesions as the basis of diagnosis and differential diagnosis, and among these patterns, liver tissue with "normal appearance" requires careful observation of easily overlooked pathological changes, so as to reduce misdiagnosis or missed diagnosis. This article mainly introduces the thoughts in the pathological diagnosis of liver tissue with normal appearance and the key points in differential diagnosis.
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Objective:To investigate the expression and significance of serum soluble T cell immunoglobulin-domain and mucin-domain protein-3 (sTIM-3) and galectin-9 (Gal-9) in patients with early acute pancreatitis (AP), so as to provide theoretical and clinical evidence for the early prediction and diagnosis of AP.Methods:From 15 September 2020 to 23 July 2021, a total of 94 AP patients with a time from onset to admission ≤48 h who were admitted to Changzhou No.2 People′s Hospital, Nanjing Medical University were selected, including 42 cases of mild acute pancreatitis (MAP), 35 cases of moderately severe acute pancreatitis (MSAP) and 17 cases of severe acute pancreatitis (SAP). The basic clinical features of AP patients were collected. Acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ), modified computed tomography severity index (MCTSI) and bedside index for severity in acute pancreatitis (BISAP) scores were evaluated in all AP patients. The levels of serum interleukin (IL)-6, IL-10, Gal-9 and sTIM-3 were detected with enzyme linked immunosorbent assay. Kruskal-Wallis test and Mann-Whitney U test were used for statistical analysis. Spearman rank correlation test and Pearson correlation analysis were used to analyze the correlation of sTIM-3, Gal-9 with inflammatory indicators and AP related scoring systems. Receiver operating characteristic curve (ROC) was performed for efficiency analysis of the combination of sTIM-3 and Gal-9 in predicting the severity of AP patients. Results:Serum sTIM-3, Gal-9 and IL-6 levels of SAP patients were higher than those of MAP patients (2 085.00 ng/L (1 628.00 ng/L, 2 673.00 ng/L) vs. 746.10 ng/L (514.50 ng/L, 1 303.00 ng/L); 466.60 ng/L (375.90 ng/L, 629.30 ng/L) vs. 108.10 ng/L (90.29 ng/L, 138.90 ng/L); (323.60±62.93) ng/L vs. (42.90±28.82) ng/L), while IL-10 level was lower than that of MAP patients ((760.30±200.40) ng/L vs. (1 206.00±566.30) ng/L), and the differences were statistically significant ( Z=45.00 and <0.01, t=23.62 and 3.15; all P<0.01). The APACHE Ⅱ and BISAP scores of SAP patients were higher than those of MAP and MSAP patients (12.00(6.00, 16.50) vs. 3.00(2.00, 5.00) and 6.00(3.00, 8.00); 3.00(3.00, 4.00) vs.1.00(1.00, 1.00) and 2.00(2.00, 3.00)), and the MCTSI score was higher than that of MAP patients (4.00(3.00, 6.00) vs. 2.00(0.00, 2.00)), and the differences were statistically significant ( Z=644.50, 704.00, 474.50, 492.50 and 664.00, all P<0.001). Serum sTIM-3 and Gal-9 were positively correlated with the pro-inflammatory factor IL-6 ( r=0.552 and 0.297, P<0.001 and =0.004). Serum sTIM-3 was negatively correlated with the anti-inflammatory factor IL-10 ( r=-0.397, P<0.001). There was no correlation between Gal-9 and the anti-inflammatory factor IL-10 ( P>0.05). Serum sTIM-3 and Gal-9 were positively correlated with APACHE Ⅱ, MCTSI and BISAP scores ( r=0.210, 0.271 and 0.363, P=0.042, =0.008 and <0.001; r=0.390, 0.448 and 0.440, all P<0.001). The areas under ROC curves (95% confidence interval) of serum sTIM-3 and Gal-9 detected alone and in combination was 0.805 (0.716 to 0.895), 0.814 (0.725 to 0.903) and 0.856 (0.773 to 0.939), respectively, and the sensitivity was 69.2%, 67.3%, 75.0%, respectively, and the specificity was 83.3%, 97.6%, 97.6%, respectively. Conclusions:The serum levels of sTIM-3 and Gal-9 increased in patients with early AP and are correlated with the severity of AP. The combined detection of sTIM-3 and Gal-9 has high sensitivity in predicting early AP, and the two indicators may be the reliable predictors of early AP.
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A retrospective analysis was performed on the clinical data of a case of intellectual developmental disorders with dysmorphic facies and behavioral abnormalities admitted in the Department of Neurology and Endocrinology, Children′s Hospital Affiliated to Shandong University in February 2020.The proband was a 3 years and 6 months old boy, who was hospitalized because of " convulsions for more than 1 year" . Physical examination revealed facial deformities.Gesell developmental schedule showed that adaptive and gross motor behavior development was severely retarded, and fine motor, language and personal-social behavior development was moderately retarded.Brain magnetic resonance imaging suggested schizencephaly.Electroencephalogram results indicated extensive discharges mainly in bilateral anterior head areas, and one myoclonic seizure was detected.Gene detection results disclosed the pathogenic variation of the proband, which was a heterozygote mutation (c.2480_2484del) in FBXO11 gene.High-throughput sequencing technology increases the possibility of identifying potential genetic mutations as the cause of disease.Patients with recurrent seizures, multi-malformation and general developmental delays should undergo gene detection in time to clarify the etiology.This technique can guide prenatal diagnosis and genetic counseling.
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A retrospective analysis was performed on the clinical data of a case of intellectual developmental disorders with dysmorphic facies and behavioral abnormalities admitted in the Department of Neurology and Endocrinology, Children′s Hospital Affiliated to Shandong University in February 2020.The proband was a 3 years and 6 months old boy, who was hospitalized because of " convulsions for more than 1 year" . Physical examination revealed facial deformities.Gesell developmental schedule showed that adaptive and gross motor behavior development was severely retarded, and fine motor, language and personal-social behavior development was moderately retarded.Brain magnetic resonance imaging suggested schizencephaly.Electroencephalogram results indicated extensive discharges mainly in bilateral anterior head areas, and one myoclonic seizure was detected.Gene detection results disclosed the pathogenic variation of the proband, which was a heterozygote mutation (c.2480_2484del) in FBXO11 gene.High-throughput sequencing technology increases the possibility of identifying potential genetic mutations as the cause of disease.Patients with recurrent seizures, multi-malformation and general developmental delays should undergo gene detection in time to clarify the etiology.This technique can guide prenatal diagnosis and genetic counseling.
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Objective@#To investigate the effect of a comprehensive exercise intervention program combined with diet control on ANGPTL2 and vascular endothelial function in obese male adolescents, and to provide theoretual basis for exercise to reduce the risk of cardiovascular disease in obese male adolescents.@*Methods@#Forty two obese male adolescents and 10 healthy male adolescents were selected, a comprehensive intervention of 6 weeks of exercise combined with diet control was carried out on obese male adolescents, and changes in morphological function, glucose and lipid metabolism, inflammation level and vascular endothelial function were measured before and after the intervention.@*Results@#After 6 weeks of comprehensive intervention, the morphological and functional indicators of obese adolescents were significantly reduced:weight, BMI, waist circumference, waist to hip ratio, systolic blood pressure, diastolic blood pressure ( P <0.05); blood lipid levels significantly decreased, including TC, TG and LDL-C ( P <0.05); inflammation levels significantly decreased, including TNF α and ANGPTL2 ( P <0.05); Vascular endothelial function significantly improved:RHI, NO/ET-1, NO, AI( P <0.05). Before and after 6 weeks of comprehensive intervention, the ANGPTL2 of obese adolescents was significantly correlated with weight ( r =0.37), BMI ( r =0.45), RHI ( r =-0.46), NO/ET-1 ( r =-0.41), NO( r =-0.45), and AI ( r =0.33)( P <0.05).@*Conclusion@#The comprehensive intervention can effectively improve the morphological function, blood lipid level and vascular endothelial function of obese male adolescents, and reduce the circulating level of ANGPTL2. ANGPTL2 may be involved in the process of comprehensive intervention to improve the vascular endothelial function of obese adolescents.
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The data of a child with early-onset epileptic encephalopathy in Qilu Children′s Hospital of Shandong University in February 2020 were analyzed retrospectively.The child was a 4-month-old girl, who was admitted to the hospital because of " repeated convulsions for 4 months and feeding difficulty for 1 month" at the age of 4 months.The patient suffered from epilepsy 1 day after birth, and the epilepsy type was tonic seizures.Severe developmental retardation was observed in the patient.Electroencephalogram showed multifocal discharge, which then turned to hypsarrhythmia.The cranial imaging was negative.Feeding difficulty occurred at the age of 3 months.The genetic testing revealed a de novo heterozygous missense mutation in the FGF12 gene (Arg114His). Various antiepileptic drugs and ketogenic diet were ineffective.There was no attack in 2 months after adding Phenytoin.The child could eat on her own after seizure control, but there was no progress in intellectual and motor development.Mutations in the FGF12 gene lead to poor prognosis of early-onset epileptic encephalopathy, and the seizures are difficult to control.Sodium ion channel blockers such as Phenytoin should be used as soon as possible.
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Polysaccharide from traditional Chinese herb, Saposhnikovia divaricata (Turcz.) Schischk. (SD) was extracted, fractionated and characterized in this work. Four fractions were prepared. Their molecular weight, monosaccharide compositions, linkage modes and structural properties were characterized with SEC-MALS-RI, HPAEC-PAD, GC-MS and NMR. SDP1 was assigned as a 1, 4-α-glucan with small amount of O-6 linked branches. SDP2 contained a big amount of the 1, 4-α-glucan and a small amount of arabinogalactan, while SDP3 possessed relatively lower amount of the 1, 4-α-glucan and a big amount of the arabinogalactan. SDP4 was defined as a pectic arabinogalactan. Four fractions showed antioxidant activities in both molecular and cellular levels and their activity was ranked as SDP4 ≈ SDP3>SDP2>SDP1. The 1, 4-α-glucan in SDP1 had the weakest, while SDP3 and SDP4 showed similar and the highest antioxidant activity. The arabinogalactan was the major component of both SDP3 and SDP4, which significantly contributed to the antioxidant activity of SDP.
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OBJECTIVE@#To examine the expression of myofibroblast in gingival after orthodontic loading.@*METHODS@#Eight patients were selected as experimental group and treated with orthodontic force for 4 months. Ten patients were selected as the control group, were not loaded. The gingival protein expressions of collagen typeⅠ, collagen type Ⅲ, α-smooth muscle actin (α-SMA) were evaluated by immunohistochemistry method.@*RESULTS@#Positive expressions of collagen typeⅠ, collagen type Ⅲ were founded, while no positive staining for α-SMA in the gingival tissue except vascular epithelium before loading. In experimental group, collagen type I and collagen type Ⅲ were increased after orthodontic loading (P<0.05), the expression of α-SMA was detected and statistically significant (P<0.05).@*CONCLUSIONS@#The myofibroblast exists in gingival tissue after orthodontic loading, and it may be concerned with orthodontic teeth relapse.
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Humans , Actins , Cell Differentiation , Collagen , Collagen Type I , Fibroblasts , Gingiva , MyofibroblastsABSTRACT
OBJECTIVE@#To explore the expression patterns of transcription factor SOX12 in lung adenocarcinoma and its significance in the diagnosis and prognosis of the malignancy.@*METHODS@#Large cancer genome databases were used to analyze SOX12 expression level in lung adenocarcinoma. Immunohistochemistry (IHC) and semiquantitative PCR were used to detect the expression of SOX12 in 36 specimens of lung adenocarcinoma tissues, 15 adjacent tissues and 21 normal lung tissues. The prognostic value of SOX12 in lung adenocarcinoma and lung squamous cell carcinoma were analyzed using Kaplan-Meier Plotter database, and the relationship between SOX12 expression and the overall survival (OS) and progression free survival (PPS) of the patients were analyzed.@*RESULTS@#Analysis of TCGA database and GEO (GSE40419) database showed that SOX12 expression levels were significantly higher in in lung adenocarcinoma than in normal lung tissues ( < 0.001). The results of IHC and semiquantitative PCR revealed that SOX12 was expressed at significantly higher levels in lung adenocarcinoma than in normal lung tissues ( < 0.001). Kaplan-Meier survival analysis showed that patients with lung adenocarcinoma positive for SOX12 had a significantly shorter OS and PPS time than those negative for SOX12 ( < 0.05), but SOX12 positivity did not significantly affect OS and PPS of patients with lung squamous cell carcinoma.@*CONCLUSIONS@#High expression levels of SOX12 in lung adenocarcinoma are significantly associated with a poor OS of the patients, suggesting the value of SOX12 to assist in early diagnosis and prognostic evaluation of lung adenocarcinoma.
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Humans , Adenocarcinoma , Metabolism , Adenocarcinoma of Lung , Metabolism , Mortality , Biomarkers, Tumor , Metabolism , Carcinoma, Non-Small-Cell Lung , Metabolism , Mortality , Databases, Factual , Kaplan-Meier Estimate , Lung Neoplasms , Metabolism , Mortality , Prognosis , SOXC Transcription Factors , Metabolism , Transcription FactorsABSTRACT
Objective@#To investigate the effects of exercise combined with dietary intervention on glucose and lipid metabolism, liver and kidney function and cardiovascular function in obese adolescents.@*Methods@#Seventy-one obese adolescents were enrolled in this study. Six weeks of exercise combined with dietary intervention were performed to determine anthropometry, glucose and lipid metabolism, liver and kidney function and cardiovascular function before and after intervention.@*Results@#After 6 weeks of comprehensive intervention, in addition to significant improvement in anthropometry indicators, fasting insulin [(18.76±11.46 vs 11.32±6.54)uU/L], LDL-C [(2.96±0.69 vs 2.22±0.62)mmol/L], TG [(1.57±0.82 vs 0.89±0.37)mmol/L] and TC [(4.52±0.76 vs 3.53±0.62)mmol/L] decreased significantly and insulin resistance improved significantly. AST [(36.1±32.28 vs 22.89±7.27)U/L], ALT [(57.42±61.25 vs 27.86±22.12)U/L], blood urea nitrogen [(4.78±0.89 vs 3.44±0.79)mmol/L] and uric acid [(498.83±120.6 vs 471.07±120.96)mmol/L] were significantly decreased and the detection rates of fatty liver and kidney were significantly decreased(P<0.05), RHI(1.34±0.28 vs 1.69±0.45) and nitric oxide/ endothelin-1 (ET-1) (1.69±0.41 vs 2.67±0.86) significantly increased and atherogenic index (3.06±0.96 vs 2.58±0.80) significantly decreased(P<0.01).@*Conclusion@#The comprehensive intervention of exercise combined with diet can effectively improve anthropometry indicators, glycolipid metabolism, liver and kidney function as well as cardiovascular function of obese adolescents.
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Aim To investigate the effects of chronic corticos-terone injection on anxiety and depression-like behavior of tree shrews, evaluate the predictability of drug and establish a novel animal model of anxious depression .Methods Twelve Chinese and Burma tree shrews were randomly divided into normal group, model group and venlafaxine group .The anxious depres-sion model of tree shrew was established by chronic corticoster-one injection ( ih, 27 mg· kg-1 , 21 d) .The venlafaxine group received intragastric administration (6 mg· kg-1).Autonomous activity score, sugar water preference test and Morris water maze test were used to evaluate the anxiety and depression-like behav-ior of tree shrews .The expressions of CRH , ACTH and COR in the tree shrew plasma were determined by Elisa kit .The con-tents of monoamine neurotransmitters of tree shrews in the hippo-campus , amygdala and prefrontal cortex were detected by HPLC-ECD.Results Compared with the normal group , the autono-mous activity score , sugar water partial eclipse degree and the learning and memory ability significantly decreased (P<0.01), while the contents of CRH , ACTH and COR significantly in-creased ( P<0.05) , and the contents of 5-HT, NE and DA in the hippocampus , amygdala and prefrontal cortex declined in the model group(P<0.05).In the venlafaxine group, the learning and memory abilities of the tree shrews were improved , the lev-els of CRH and COR in plasma were significantly decreased ( P<0.05), and the contents of 5-HT, NE and DA were increased (P<0.05).Conclusions The tree shrews of anxious depres-sion have obvious HPA axis hyperactivity and monoamine neuro-transmitter disorder , and venlafaxine can reverse this phenome-non, indicating that the tree shrews model of anxious depression has drug predictability , which is a kind of novel animal model of anxious depression closer to human in clinic .
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Purpose To investigate the difference of expression of autophagy-related gene (Beclin1,LC3,mTOR) in the development of esophageal squamous cell cancer.Methods Immunohistochemical EnVision method was adopted to detect the expression of autophagy-related gene Beclinl,LC3 and mTOR in 30 cases of normal esophageal mucosa,32 cases of low-grade intraepithelial neoplasia (LGIN),34 cases of highgrade intraepithelial neoplasia (HGIN),35 cases of early carcinoma and 126 cases of advanced esophageal carcinoma,respectively.The correlation between their expression with clinicopathologic factors was also analysed.Results The expression of Beclin1 in advanced esophageal carcinoma was obviously higher than that in another four groups (P ＜ 0.005).LC3 expression in advanced esophageal carcinoma was significantly higher than that in normal esophageal mucosa,LGIN and early carcinoma (P ＜ 0.005).The expression of mTOR in advanced esophageal carcinoma was significantly higher than that in normal esophageal mucosa,LGIN and HGIN (P ＜ 0.005).In advanced esophageal carcinoma group,the expression of Beclin1,LC3 and mTOR was related to tumor TNM stage and lymph node metastasis (P ＜ 0.05).Beclin1 expression was positively associated with LC3 and mTOR expression in advanced squamous cell carcinoma (P ＜ 0.05).Positive correlation was also observed between the expression of mTOR and LC3 in advanced esophageal carcinoma and HGIN (P ＜ 0.05).Conclusion In the carcinogenesis and development of esophageal cancer,Beclin1,as a tumor suppressor gene,activates autophagy and leads to excessive self consumption and death of tumor cells.mTOR promotes tumor growth by inhibiting autophagy and promoting angiogenesis.The combined detection of Beclinl,LC3 and mTOR may be beneficial to evaluate the progression and prognosis of esophageal squamous cell cancer.
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We investigated the epidemiological and clinical characteristics,to provide evidence for the control and prevention of Creutzfeldt-Jakob disease (CJD) in Shaanxi Province,China.Clinical and epidemiological datas on 49 suspicious CJD patients from 7 hospitals in Shaanxi Province from 2011 to 2015 was collected.Blood and cerebral spinal fluid (CSF) specimens from the cases were gathered.For CSF sample,14-3-3 protein were tested by Western blot.For blood sample,PCR and sequencing were used,and then the mutation of PRNP gene and the polymorphism of 129 and 219 amino acids were analyzed.It showed that a total number of 16 probable and 4 possible sporadic CJD patients,and 1 familial CJD case were identified.Among these cases,neither geographic nor occupational-related ones were found.The median age of onset for the probable sporadic CJD cases was 62 years old,and the gender ratio of male to female was 1.29 to 1.The most initial symptom was rapid progressive dementia,which accounted for 47.62％ of the CJD patients.This report indicates that the main type of CJD in Shaanxi Province is sporadic CJD with its distinctive characteristics including geography distribution,occupation,gender ratio and age of onset.
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AIM:To explore the effects of chloroquine (CQ) on collagen Ⅰ and collagen Ⅲ expression in activated rat hepatic stellate cell line HSC-T6 and the possible mechanism.METHODS:Transforming growth factor-β1 (TGF-β1) was used to activate HSC-T6 cells and 3 doses of CQ was administered for 24 h.The cells were divided into 5 groups as follows:control group,TGF-β1 group,TGF-β1 + CQ (15 μmol/L) group,TGF-β1 + CQ (30 μ mol/L) group and TGF-β1 + CQ (60 μmol/L) group.Western blot was used to determine the expression of LC3-Ⅱ/LC3-Ⅰ,P62 and o-SMA in activated HSC-T6 cells.The expression of collagen Ⅰ and collagen Ⅲ was detected by immunocytochemical staining,Western blot and RT-qPCR.Western blot and RT-qPCR were also used to detect the expression of matrix metalloproteinase-13 (MMP-13),tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 at mRNA and protein levels.RESULTS:The ratio of LC3-Ⅱ / LC3-Ⅰ and P62 expression were increased after CQ intervention.Moreover,they were significantly higher in the TGF-β1 + CQ groups than those in TGF-β1 group (P ＜ 0.01).The expression of collagen Ⅰ and collagen Ⅲ at mRNA and protein levels was significantly increased in all TGF-β1 + CQ groups as compared with TGF-β1 group (P ＜0.01),and it was markedly increased among TGF-β1 + CQ groups in a dose-dependent manner.The expression of MMP-13 at mRNA and protein levels was significantly lowered and that of TIMP-1 and TIMP-2 was significantly increased in TGF-β1 + CQ groups as compared with TGF-β1 group (P ＜0.05).CONCLUSION:Inhibition of autophagy by CQ in activated HSC-T6 cells up-regulates the expression of collagen Ⅰ and collagen Ⅲ in a dose-dependent way,probably due to reduction of MMP-13 and enhancement of TIMP-1 and TIMP-2 expression.
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AIM:To explore the effects of chloroquine (CQ) on collagen Ⅰ and collagen Ⅲ expression in activated rat hepatic stellate cell line HSC-T6 and the possible mechanism.METHODS:Transforming growth factor-β1 (TGF-β1) was used to activate HSC-T6 cells and 3 doses of CQ was administered for 24 h.The cells were divided into 5 groups as follows:control group,TGF-β1 group,TGF-β1 + CQ (15 μmol/L) group,TGF-β1 + CQ (30 μ mol/L) group and TGF-β1 + CQ (60 μmol/L) group.Western blot was used to determine the expression of LC3-Ⅱ/LC3-Ⅰ,P62 and o-SMA in activated HSC-T6 cells.The expression of collagen Ⅰ and collagen Ⅲ was detected by immunocytochemical staining,Western blot and RT-qPCR.Western blot and RT-qPCR were also used to detect the expression of matrix metalloproteinase-13 (MMP-13),tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 at mRNA and protein levels.RESULTS:The ratio of LC3-Ⅱ / LC3-Ⅰ and P62 expression were increased after CQ intervention.Moreover,they were significantly higher in the TGF-β1 + CQ groups than those in TGF-β1 group (P ＜ 0.01).The expression of collagen Ⅰ and collagen Ⅲ at mRNA and protein levels was significantly increased in all TGF-β1 + CQ groups as compared with TGF-β1 group (P ＜0.01),and it was markedly increased among TGF-β1 + CQ groups in a dose-dependent manner.The expression of MMP-13 at mRNA and protein levels was significantly lowered and that of TIMP-1 and TIMP-2 was significantly increased in TGF-β1 + CQ groups as compared with TGF-β1 group (P ＜0.05).CONCLUSION:Inhibition of autophagy by CQ in activated HSC-T6 cells up-regulates the expression of collagen Ⅰ and collagen Ⅲ in a dose-dependent way,probably due to reduction of MMP-13 and enhancement of TIMP-1 and TIMP-2 expression.
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Aim To explore the proteomics mechanism of the differentiation induction effect of 4-amino-2-trif-luoromethyl-phenyl retinate(ATPR)on human leukemi-a K562 cells. Methods Human leukemia K562 cells were incubated with the same concentration (1 × 10 - 6 mol·L - 1 ) of ATPR or ATRA for 48 hours. The total cell proteins were collected, purified and digested by trypsin, solid phase extraction, and the peptides were detected by ESI-LC-MS / MS. The difference of the pro-tein expression between the cells treated with ATPR and ATRA was compared by using the Discoverer Pro-teome 1. 2 software, and the molecular function, the biological process and other information of those pro-teins were analyzed based on the DAVID, KEGG, STRING databases. Results 120 specific proteins were identified only in the ATPR group, 143 only in the ATRA group, and 422 other proteins in both groups. Results of DAVID analysis showed that ATPR-induced specific proteins were mainly involved in 39 biological processes of proteins and macromolecules metabolism, protein transport and localization and so on. Results of KEGG analysis revealed that ATPR-in-duced proteins participated in signal pathways, mainly metabolic pathways, PI3K-Akt signal pathway, TGF-beta signal pathway and other pathways in cancer. String protein interaction network analysis displayed that ATPR-induced proteins, like EIF3A, EIF6, RPL3, RPL8, RPL13, RPL7A, RPL21, RPS3, RPS14, NACA, BTF3, NHP2L1, PPP2CA proteins had direct interactions with more than or equal to 10 associated proteins. Conclusion The differentiation induction effect of ATPR on K562 cells might be as-cribed to the ATPR-induced proteins interaction net-work and the specific central proteins it induced, which are involved in the regulation of cell prolifera-tion, differentiation and apoptosis.
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Due to the weak foundation of clinical trials,the pre-market data of pediatric medicine is insufficient. Therefore,a comprehensive evaluation system of pediatric clinical medicine is urgently needed to safeguard the children′s medication safety. This article provides the method and builds path to establish the comprehensive evaluation system from the aspects of biulding organization?al structure,operational process and evaluation methodology,respectively. Further,based on the concept ofEvidence-based or other?wise evidence-created,and following the example of“Mini-Sentinel”of the United States,the big data in medicine can be used to overcome the“Evidence-created”difficulty in the evaluation of pediatric medication. Thus more extensive comprehensive evaluation can be made through collecting the clinical data generated from the real world for the pediatric medication. In the future ,the compre?hensive evaluation system of pediatric clinical medicine established on the basis of big data can be linked to thePrecision MedicineProject in our country,the evaluation bases can be the Mini-Sentinel to collect big medical data and biological samples,thus to bring a steady flow of power for promoting the pediatric medicine active surveillance and pediatric precision medicine ,and finally to safeguard the children′s health in our country.
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<p><b>OBJECTIVE</b>To investigate the functions of human periodontal myofibroblast (MFB) in vitro.</p><p><b>METHODS</b>Human periodontal fibroblast (hPDLFs) was cultured and induced to MFB by transforming growth factor-β1 (TGF-β1). MFB was denoted as the experimental group, whereas the hPDLFs was the control group. The groups were continuously cultured and harvested at 0, 12, 24, 48, and 72 h. The MFB marker α-smooth muscle actin (α-SMA) was examined by immunocytochemistry. The expression of fibronectin (FN) between MFB was examined by immunocytochemistry to detect the MFB contact relationship. The mRNA expression levels of α-SMA, collagen (Col) I, and Col III were measured by reverse transcription-polymerase chain reaction (RT-PCT) to analyze extracellular matrix secretion. The protein expression levels of α-SMA and Col I were also assessed by Western blot.</p><p><b>RESULTS</b>The experimental group had significantly higher α-SMA expression than the control group at 0 h (P < 0.001). A positive expression of FN was found between MFB. The experimental group had significantly higher expression levels of Col I and Col III than the control group at 24 h (P < 0.001).</p><p><b>CONCLUSION</b>Human periodontal MFB presents a continuous, high expression of α-SMA. MFB could interact through FN. MFB is significantly capable of extracellular matrix secretion.</p>
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Humans , Actins , Epithelial Cells , Extracellular Matrix , Fibroblasts , Fibronectins , Jaw , Metabolism , Myofibroblasts , Transforming Growth Factor beta1ABSTRACT
Objective To prepare dual?modality single?photon emission computed tomography (SPECT)?MRI molecular nanoprobes targeting HAb18G/CD147 expressed on breast cancer cell membranes and investigate the physicochemical and biological properties in vitro. Methods Superparamagnetic iron oxide nanoparticles (SPIOs) were prepared by one?pot reaction method as described. The single?chain antibody fragments HAb18F(ab')2 were conjugated to SPIOs via chemical method and then labeled with 125I using Iodogen method. The final 125I?SPIO?HAbF18(ab')2 nanoprobes were purified. SPIOs or 125I?HAb18F(ab')2 were used as control. We carried preliminary evaluation on their physicochemical properties and biological characteristics in vitro: transmission electron microscope (TEM) and dynamic light scattering (DLS) were used to measure these nanoparticle sizes and the hydrodynamic diameters. The MRI T2 transverse relaxation efficiency of these nanoprobes at different Fe2+concentrations were measured with 1.5 T clinical MR scanner. The 125I?SPIO?HAb18F(ab')2 and 125I?HAb18F(ab')2 radiochemical purity were measured by thin layer chromatography and the radio chemical yield was calculated. We also conducted stability tests in vitro and octanol/water partition coefficient experiments. Two breast tumor cell lines, MDA?MB?231 (HAb18G?overexpressing cells,experimental group) and MDA?MB?468 (control), were used for assessment of cells viability at different Fe2 + concentrations (1, 5, 10, 20, 40 μg/ml) by methyl thiazolyl tetrazolium assay. Specific binding experiments in vitro included two parts:magnetic resonance imaging and radionuclide tests, the above?mentioned breast cancer cell lines were incubated with 125I?SPIO?HAb18F(ab')2 nanoprobes respectively and took MDA?MB?231 cells which were not treated as blank group. First comparing the MR signal intensity differences among experimental group, the control group and blank group, then calculated the rate of MRI signal changes;Two breast tumor cell lines, MDA?MB?231 and MDA?MB?468 were incubated with 125I?SPIO?HAb18F(ab')2 nanoprobes too, then measured radioactivity counting byγcounter at different time and calculated the cell binding rates, and did statistical analysis by using one?way ANOVA. Results The SPIOs were fairly homogeneous with an average core size of (10.32±1.30) nm;the SPIO and 125I?SPIO?HAb18F(ab')2 hydrodynamic diameter of 44.80 and 52.64 nm, and MRI scanning showed that the transverse relaxation efficiency of SPIO and 125I?SPIO?HAb18F(ab')2 were 38.79 and 106.73 mM-1 · s-1, respectively. The radio chemical yield of 125I?SPIO?HAbF18(ab')2 and 125I?HAb18F(ab')2 were 41.90% and 85.50%, respectively. The radio chemical yield of the two groups were >95%, suggesting well stability in vitro. The lipo?hydro partition coefficient values were -0.99 ± 0.03 and-1.49 ± 0.08, respectively, which demonstrated that they were both water?soluble substances. Different Fe2+concentrations (1,5,10,20,40μg/ml) of 125I?SPIO?HAb18F(ab')2 on breast cancer cell lines MDA?MB?231 and MDA?MB?468 showed no significant inhibition of cell proliferation (F values were 0.78, 0.66; P values were 0.58, 0.66). The cell?specific binding experiment showed: MRI signal intensity values on experimental group, the control group and the blank group were (1 670 ± 5), (1 930 ± 8), (2 349 ± 14), respectively, significant differences existed among these groups (F=4 408.48,P=0.000), the rate of signal intensity change of experimental group and the control group were 28.87%,17.78%. SPECT:MDA?MB?231 could uptake 125I?SPIO?HAb18F(ab')2, the cell binding rates were (6.52 ± 0.60)% and (10.52 ± 2.04)% in 20 min and 4 h, respectively.Conclusions Our results suggested that the dual?modality SPECT?MRI nanoprobes 125I?SPIO?HAb18F(ab')2 were prepared successfully with good physicochemical properties and biological characteristics in vitro. These dual?modality molecular imaging nano?probes may have potential to improvearly detection and diagnosis of HAb18G/CD147?expressing cancers and to facilitate the development of HAb18G/CD147?directed interventions.
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An important index determination for clinical diagnosis of renal function is to assay the creatinine concentration in serum. In the analytical process applied with coupled-enzyme, the quality control of sarcosine oxidase (SOX) as a key enzyme is the first problem to be solved. In order to establish an efficient and laboratory-scale production of SOX, the recombinant sarcosine oxidase (r-SOX) gene was a high-level expression in E. coli induced with lactose on a large-scale fermentation in 300 L fermenter. The results suggested that the biomass concentration reached OD600 of 22 and the expression of recombinant sarcosine oxidase in E. coli accounted for about 25% of total soluble protein in culture after fermentation. The cell-free extract obtained from high pressure homogenizer was processed by selective thermal denaturation and then purified with Ni-Sepharose FF chromatography. The sarcosine oxidase with 97% purity, 25 U/mg specific activity and 92.4% activity recovery was obtained. The molecular weight with single peptide chain of 53 kD and 55 kD of recombinant sarcosine oxidase was assessed by SDS-PAGE in presence or absence of 2-mercaptoehanol and Sephacryl S-200 chromatography. This sarcosine oxidase was found to be a conjugated protein, yellow enzyme, which combined with FAD as prosthetic group by covalent linkage. The contaminant of catalase was not detected in the sample pool of this enzyme. In addition, a further test to the thermal stability of sarcosine oxidase was done. According to the above results, the development and utilization of this enzyme has been set up on a reliable foundation.