Characteristics and problems of hydroxyapatite/polymer bone repair material / 中国组织工程研究

BACKGROUND:Hydroxyapatite is the main inorganic component of bone tissue.The polymer has the structure and function of a biomimetic extracellular matrix.The composites of hydroxyapatite and polymer have been widely studied. OBJECTIVE:To summarize the research status of hydroxyapatite composite polymer materials for bone tissue repair. METHODS:The articles collected in PubMed,Web of Science,CNKI and WanFang databases were searched from January 2010 to April 2023.The Chinese and English search terms were"hydroxyapatite,polymer,composites,degradability,bone defect,bone repair".Finally,75 articles were included for review. RESULTS AND CONCLUSION:Polymers often used in composite with hydroxyapatite for bone tissue repair include natural polymers(collagen,chitosan,alginate,serine protein,cellulose,hyaluronic acid,and polyhydroxybutyrate)and synthetic polymers[polylactic acid,polylactic acid-hydroxyacetic acid copolymer,poly(has-lactide),poly(amino acid)and poly(vinyl alcohol)].The mechanical properties and osteoinductivity of hydroxyapatite/polymer composites were improved compared with pure hydroxyapatite.Hydroxyapatite composite with polymers can be made into porous scaffolds,hydrogels,and coatings for bone repair.Hydroxyapatite/polymer composites can accelerate bone reconstruction with a slow release of loaded drugs and cytokines due to their bionic extracellular matrix structure and function.Based on the diversity of causes of bone defects and the fact that bone repair is a complex continuous process involving multiple biological factors and proteins,repair materials with mechanical properties matching bone tissue,degradation processes synchronized with bone repair,and efficient osteogenesis and vascularization need to be further investigated.

The mechanism of methotrexate in the treatment of rheumatoid arthritis using untargeted metabolomics / 中华风湿病学杂志

Objective:To investigate the mechanism of methotrexate in the treatment of rheumatoid arthritis (RA) by constructing a rat model of collagen-induced arthritis (CIA) and using non-targeted metabolomics.Methods:Enzyme-linked immunosorbent assay (ELISA) was used to determine the contents of TNF-α, IL-1β, IL-6, IL-4 and IL-10 in serum. HE staining and Masson staining were used to observe the histological changes of joints in each group. Non-targeted gas chromatography-mass spectrometry metabolomics technique was used to screen the expression profiles of differential metabolites in serum and cluster analysis and KEGG enrichment analysis were performed to screen the differential metabolic pathways, and real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the key enzymes in the differential metabolic pathways. All experimental data conforming to the normal distribution were compared between groups using one-way ANOVA, and P<0.05 was considered statistically significant. Results:MTX significantly improved the joint inflammatory response and arthritis score and increased the body weight of CIA rats. The results of HE and Masson staining showed that MTX could ameliorate the erosion of articular cartilage by synovial tissue in CIA rats. ELISA results showed that MTX significantly decreased the contents of TNF-α [(191.2±17.4)pg/ml, F=40.31, P<0.001], IL-1β[(28.4±1.2)pg/ml, F=10.11, P=0.012] and IL-6[(118.7±1.4)pg/ml, F=829.40, P<0.001] in the serum and increased the contents of IL-4 [(49.3±3.3)pg/ml, F=33.44, P<0.001] and IL-10 [(30.2±0.7)pg/ml, F=33.44, P<0.001] in the serum of CIA rats. Non-targeted metabolomics technique showed MTX had an effect on metabolites such as phosphocholine, palmitic acid, oleic acid, and choline in the serum of CIA rats. KEGG pathway enrichment analysis showed that MTX had an effect on glycerophospholipid metabolism( P<0.01)and sphingolipid metabolism( P<0.05)in CIA rats. qRT-PCR results showed that MTX could down-regulate the expression of the key enzymes such as Plb1 [(1.00±0.49), F=8.23, P=0.019], Gpcpd1[(1.10±0.09), F=8.19, P=0.019], Chka [(1.33±0.19), F=33.00, P<0.001], Chkb [(2.07±1.21), F=8.20, P=0.019]and Phospho1 [(1.07±0.14), F=13.58, P=0.006]in the glycerophospholipid metabolic pathway in the synovial membrane of CIA rats, and can also down-regulate the expression of the key enzymes Kdsr [(1.24±0.32), F=13.85, P=0.006], Plpp1 [(1.61±0.32), F=11.95, P=0.003) and Degs1 [(1.21±0.15, F=46.55, P<0.001]in the sphingolipid metabolic pathway. Conclusion:The biological mechanism of MTX in the treatment of rheumatoid arthritis may be related to the down-regulation of glycerophospholipid metabolism and sphingolipid metabolism pathway metabolic levels in the body.

Stimulation quantitation of saponins and sterones from Achyranthis Bidentatae Radix by high-performance liquid chromatography with double external standards calibration method / 药学学报

Saponins and sterones are two main characteristic components in Achyranthis Bidentatae Radix. In order to control the quality of Achyranthis Bidentatae Radix more effectively, a high-performance liquid chromatography (HPLC) method was established by using double external standards calibration method (DESCM) for simultaneous determination of the contents of achyranthoside C, achyranthoside D, β-ecdysterone, 25R-inokosterone and 25S-inokosterone in Achyranthis Bidentatae Radix. Chromatographic separation was achieved on an Agilent Poroshell 120 EC-C18 (150 mm × 4.6 mm, 2.7 µm) using 0.1% phosphoric acid in water and 0.1% phosphoric acid in acetonitrile as mobile phase. The flow rate was 0.8 mL·min-1 and the column temperature was set as 35 ℃, the injection volume was 5 μL and the total analytical time was 30 min. β-Ecdysterone was used as the reference to calculate the relative correction factors (RCF) and relative retention time (RRT) of 25R-inokosterone and 25S-inokosterone, achyranthoside D was used for achyranthoside C. The RCFs of 25R-inokosterone, 25S-inokosterone, and achyranthoside C were 1.116, 1.056, and 0.888 1, respectively. The double external standards calibration method (DESCM) and external standard method (ESM) were used to calculate the contents of five ingredients in Achyranthis Bidentatae Radix samples from different sources and the variation between the results was within acceptable limits (RE ≤ 5%). The results showed that the contents of two saponins and three sterones of Achyranthis Bidentatae Radix were 0.597%-1.916% and 0.044%-0.150% respectively. The total content of saponins was about 10 times that of sterones. In conclusion, the established DESCM allowed simultaneous determination of five ingredients (achyranthoside C, achyranthoside D, β-ecdysterone, 25R-inokosterone, and 25S-inokosterone) in Achyranthis Bidentatae Radix, providing a scientific and feasible overall quality evaluation method for Achyranthis Bidentatae Radix.

A new dimeric benzylisoquinoline alkaloid from Corydalis yanhusuo / 药学学报

(±)-Bicoryanhunine B (1), a new dimeric benzylisoquinoline alkaloid was isolated from the dried tubers of Corydalis yanhusuo by various chromatographic methods, including silica gel, Sephadex LH-20, reverse phase C18, and semi-preparative HPLC. Its structure was determined by spectroscopic methods, including UV, IR, ESI-MS, HR-ESI-MS and 1D/2D NMR. (±)-Bicoryanhunine B (1) was a moderate PD-1/PD-L1 interaction inhibitor with an IC50 value of 7.80 ± 0.49 μmol·L-1. In addition, 1 exhibited potent inhibitory activities against LPS-induced NO production in RAW 264.7 macrophages with an IC50 value of 4.83 ± 2.21 μmol·L-1.

Physical and neuropsychological development of children with Citrin deficiency / 中国当代儿科杂志

OBJECTIVES@#To study the physical and neuropsychological development of children with Citrin deficiency (CD).@*METHODS@#A total of 93 children, aged 1.9-59.8 months, who were diagnosed with CD by @*RESULTS@#For the 93 children with CD, the incidence rate of failure to thrive was 25% (23 children) and the proportion of small for gestational age was 47% (44 children). For the 100 cases of CD, the incidence rates of growth retardation, underweight, emaciation, overweight, and microcephalus were 23% (23 cases), 14% (14 cases), 4% (4 cases), 8% (8 cases), and 9% (9 cases), respectively. The incidence rate of neuropsychological developmental delay was 25% (25 cases), and the incidence rates of development delay in the five domains of adaptability, gross motor, fine motor, language, and social ability were 7% (7 cases), 15% (15 cases), 7% (7 cases), 9% (9 cases), and 7% (7 cases), respectively.@*CONCLUSIONS@#Physical and neuropsychological developmental delay can be observed in children with CD, and physical and neuropsychological development should be regularly assessed.

Effect of Amino Acid Motifs in Integrin β3 Cytoplasmic Tail on αⅡbβ3-Mediated Cell function in 293T cell models / 中国实验血液学杂志

OBJECTIVE@#To establish 293T cell lines stably expressing Calpain-cleavage related α3 cytoplasmic tail mutants, and to explore the effect of amino acid motifs in integrin β3 cytoplasmic tail on αⅡbβ3-mediated cell function.@*METHODS@#293T cell lines stably co-expressing human wild type integrin αⅡb and full length β3 or mutant β3, including β3-ΔNITY (β3 cytoplasmic tail NITY motif deleted), β3-Δ754 (β3 cytoplasmic tail TNITYRGT motif deleted) and β3-Δ759 (β3 cytoplasmic tail RGT motif deleted) were established. Spreading and adhesion of these stable cell lines on immobilized fibrinogen were tested.@*RESULTS@#293T-αⅡbβ3ΔNITY, 293T-αⅡbβ3Δ754, 293T-αⅡbβ3Δ759 and 293T-αⅡbβ3 cell lines were successfully established. Compared with the 293T cells, 293T-αⅡbβ3 cells which expressed full β3, possessed well adhesion and spread ability on immobilized fibrinogen, suggesting it can be as a surrogate for platelet. Compared with 293T-αⅡbβ3 cells, the 293T-αⅡbβ3ΔNITY cells showed a partial impairment of adhesion and spreadability on immobilized fibrinogen. while the 293T-αⅡbβ3Δ754 cells and 293T-αⅡbβ3Δ759 cells failed to adhere or spread on immobilized fibrinogen.@*CONCLUSION@#To the cell spreading function mediated by integrin β3, RGT motif is vital, while NITY can be dispensable. These established 293T cell lines stably expressing different β3 mutants provide a solid basis for a further analysis of mass spectrometry.

Diagnosis of two cases from one family with Joubert syndrome caused by novel mutations of TCTN1 gene by whole exome sequencing / 中华医学遗传学杂志

OBJECTIVE@#To explore the pathogenesis of two fetuses from one family affected with Joubert syndrome (JS).@*METHODS@#Whole exome sequencing was employed to screen potential mutations in both fetuses. Suspected mutations were verified by Sanger sequencing. Impact of intronic mutations on DNA transcription was validated by cDNA analysis.@*RESULTS@#Two novel TCTN1 mutations, c.342-8A>G and c.1494+1G>A, were identified in exons 2 and 12, respectively.cDNA analysis confirmed the pathogenic nature of both mutations with interference of normal splicing resulting in production of truncated proteins.@*CONCLUSION@#The genetic etiology of the family affected with JS has been identified.Above findings have enriched the mutation spectrum of TCTN1gene and facilitated understanding of the genotype-phenotype correlation of JS.

Genotypic and phenotypic analysis of a patient with de novo partial monosomy 18p and partial trisomy 18q / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic cause for a patient with intellectual disability, short stature and multiple congenital anomalies, and to correlate the result with the clinical phenotype.@*METHODS@#Routine karyotyping analysis was carried out on GTG-banded metaphase chromosomes. Single nucleotide polymorphism (SNP) microarray was used to detect microdeletions or microduplications in the patient. Fluorescence in situ hybridization (FISH) was used to ascertain the origin of aberrant chromosomes.@*RESULTS@#The karyotype of the patient was 46,XY,der(18), while both of his parents had a normal karyotype. SNP array identified a 1.23 Mb deletion at 18p11.32-pter (chr18: 136 227-1 370 501, hg19) and a 33.76 Mb duplication at 18q21.1-qter (chr18: 44 250 359-78 013 728, hg19) in the patient. Above finding was confirmed by dual-color FISH with one color for 18p and another for 18q. The patient presented with some common features of 18p deletion and 18q duplication including intellectual disability and growth retardation, in addition with some features of 18p deletion including pectus excavatum, short stature and growth hormone (GH) deficiency. The patient showed progressive improvement of stature with GH therapy. Comparison of patients with previously reported dup(18q)+del(18p) recombinations suggested that, even for patients with similar breakpoints, their phenotypes have ranged from normal to severe and there were no consistent findings.@*CONCLUSION@#As aberrations involving double chromosomal segments often result in phenotypic variability, it has been difficult to correlate the genotype of our patient with his phenotype.

Immunometabolism and systemic lupus erythematosus / 北京大学学报(医学版)

Systemic lupus erythematosus (SLE) is a highly heterogeneous autoimmune disease, characterized by production of pathogenic autoantibodies and wide involvement of multiple systems. Damageofimmune tolerance and imbalance of immune homeostasis lead to the production of autoantibodies and the injuries of multiple organs and systems. In recent years, plenty of studies have identified that immunometabolism affects survival status of certain cells, also cell activation, differentiation and effector functions. Conversely, immune cells with different functions or differentiational status upregulate specific metabolic pathways to maintain their identities. In response to outer stimulations, naive immune cells differentiate into activated cells, accompanied with a series of immunometabolism changes. Therefore, abnormal immunometabolism can induce global imbalance of immune homeostasis, which further results in the initiation and development of autoimmune diseases, including SLE. Multiple abnormalities of immunometabolism have been found in patients with SLE or mouse models of lupus. Immune cells involved in the development of SLE, such as T cells, B cells, dendritic cells and macrophages present various metabolic abnormalities and pathological phenotypes. Among these cells, CD4+ T cells play predominant roles in the pathogenesis of SLE. Lots of studies demonstrated that CD4+ T cells and their subsets were in abnormal immunometabolic status,which further resulted in the development of SLE. In CD4+ T cells from patients with SLE or mouse models of lupus, both levels of glycolysis and oxidative phosphorylation are significantly higher compared with healthy controls. However,mitochondrial abnormalities, decreased ATP production and increased level of oxidative stress also have been found in these cells, which play important roles in the production of reactive oxygen intermediates and autoantibodies. Aggregated lipids rafts and increased synthesis of glycosphingolipid and cholesterol also have been observed in the CD4+ T cells from patients with SLE, leading to the abnormally elevated TCR signaling. Moreover, mechanistic target of rapamycin (mTOR) signaling is activated in the CD4+ T cells from both patients with SLE or mouse models of lupus and participate in the metabolic abnormalities of pathological CD4+ T cells. Progressive understanding of immunometabolism give us new insights of the pathogenesis of SLE and provide us with more therapeutic targets in the treatment of SLE.

Diagnostic Value of Nanomagicbeabs Sorting-time Resolved Fluoroimmuno Assay for Serum Calgizzar in Pancreatic Carcinoma / 现代检验医学杂志

Objective To detect the serum levels of calpainin (S100A11) using nanomagicbeabs sorting-time resolved fluoroimmuno assay (NMBS-TRFIA) and evaluate its diagnostic value in pancreatic carcinoma.Methods 88 patients with pancreatic carcinoma,50 patients with acute pancreatitis,10 patients with pancreatic cyst and 20 healthy controls were selected as the study subjects.The human peripheral serum blood was sorted with S100A11 antibody coupled nanomagicbeabs,and the concentration of S100A11 was detected by TRFIA method.The area under the receiver operating characteristic (ROC) curve (AUC) was used to determine the cut-off level for diagnosis of pancreatic carcinoma,in order to evaluate the sensitivity and specificity for diagnosis of pancreatic carcinoma.Results S100A11 showed a linear relationship within the range of 6.08～ 500 ng/ml using NMBS-TRFIA method,intraassay CV≤6.35％,inter-assay CV≤7.12％,and the average recovery rate was 104.7％.The serum levels of S100A11 in patients with pancreatic carcinoma,patients with acute pancreatitis and patients with pancreatic cyst were 185.53 ± 161.19,106.06±113.83 and 68.99± 47.83 ng/ml respectively.Compared with the normal control group (37.98±25.14 ng/ml),the differences were statistically significant (t=-8.065,-3.375,-2.266,all P ＜0.01).The serum levels of S100A11 in patients with pancreatic carcinoma was significantly higher than those in patients with acute pancreatitis and patients with pancreatic cyst (all P＜0.05).According to the ROC curve,ROCAUC=0.985 (95％ CI:0.972 ～ 0.997),the best cut-off level for the diagnosis of pancreatic carcinoma was 89.5 ng/ml (sensitivity 81.8 ％,specificity 67.5 ％).Conclusion NMBS-TRFIA can enrich S100A11 in serum and improve the detection sensitivity of serum S100A11,and the method is simple and easy to be popularized.Serum S100A11 has a high sensitivity and specificity in the diagnosis of pancreatic carcinoma,and is a new serum marker for the diagnosis of early pancreatic carcinoma.

Study on structural conversion of dihydrochelerythrine in different solvents / 中国中药杂志

Dihydrochelerythrine was isolated from the ethanol extract of Corydalis yanhusuo by chromatographic and recrystallization techniques. To our knowledge, this is the first report that dihydrochelerythrine was found to be unstable. The NMR, HPLC, and LC-MS were applied to monitor the structural conversion process of dihydrochelerythrine. The results showed that when dissolved in polar deuteration solvent (e.g., DMSO-₆ & MeOD), dihydrochelerythrine is directly converted to chelerythrine gradually. However, if used non-polar reagent (e.g.,CD₂Cl₂), the sample of dihydrochelerythrine undergoes the formation of pseudobase, chelerythrine, and bimolecular ether then followed by oxidation to oxychelerythrine as the major final product. Which leads to this phenomenon maybe is that the C-6 in dihydrochelerythrine is highly reactive to nucleophiles, and is easily converted to different derivatives in different solvents attributed to the solvent effect. This finding will contribute to the extraction and isolation, bioactivity screening, and quality evaluation of medicinal materials containing dihydrochelerythrine and other similar derivatives.

Diagnostic value of linked color imaging technology for Helicobacter pylori?related gastritis / 中华消化内镜杂志

Objective To evaluate the diagnostic value of linked color imaging (LCI) technology on Helicobacter pylori (HP)-related gastritis. Methods Forty patients who were diagnosed as chronic gastritis using blue laser imaging endoscopy in Shenzhen Hospital of Southern Medical University during November 2016 to June 2017 were enrolled in this study. The appearance of gastric mucosa was observed using conventional white light imaging and LCI. Biopsies were taken under white light imaging according to biopsy pathological diagnosis consensus, and the ones from abnormal reddening area were taken under LCI. 13C-urea breath test (13C-UBT) was performed in all 40 patients. The consistency between the two observation methods and final pathological diagnosis was evaluated using Kappa test, and the diagnostic consistency of the two methods was compared using Mc Nemar paired Chi-square test.Results The positive predictive value of white light imaging and LCI for prediction of HP infection was 54. 5%(6/11) and 81. 5%(22/27), respectively.The consistency between white light imaging diagnosis and final pathological diagnosis was 0. 475 (19/40), Kappa＝0. 635; the consistency between LCI diagnosis and final pathological diagnosis was 0. 875 (35/40), Kappa＝0. 741. Mc Nemar paired Chi-square test showed that the consistency between the two methods had significant difference (P＜0. 01). 13C-UBT showed that 19 patients were positive and 21 negative. Among the 19 positive patients, 1 case was diagnosed as HP negative by pathology under LCI; and among the 21 negative patients, 4 cases were diagnosed as HP negative by pathology under LCI.The consistency between pathological diagnosis and 13C-UBT was good (Kappa＝0. 751). The red-white boundary and diffuse redness of gastric mucosa were observed in 15 and 11 cases under LCI, respectively, while unobserved under white light imaging.The Wilcoxon signed ranks test showed that there was a significant difference between white light imaging and LCI on the appearance of gastric mucosa (Z＝－4. 455, P＜0. 01). Conclusion LCI is more useful for diagnosis of HP-related chronic gastritis than white light imaging.

Clinical phenotypes and genetic study of 2 cases with 22q13 deletion syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To determine the genetic etiology and clinical characteristics of 2 boys featuring development delay (DD).</p><p><b>METHODS</b>Routine chromosomal banding was performed to analyze the karyotypes of the patients and their parents. Single nucleotide polymorphism array (SNP array) analysis was employed to identify pathogenic deletion/duplication of chromosomes, and quantitative real-time PCR (qPCR) was performed to confirm the results.</p><p><b>RESULTS</b>Patient 1 showed a global developmental delay, especially impaired language development, seizures, behavioral problems belonging to the autism spectrum and mild facial dysmorphism. Patient 2 mainly presented with severely delayed speech and moderate intellectual disability, but did not have obvious facial dysmorphism and autistic-like behavior. The diagnosis of 22q13 syndrome was established based on identification of a heterozygous microdeletion at chromosome 22q13.33 in both patients (69 kb and 587 kb, respectively) by the SNP array analysis. Both patients had deletions of SHANK3 and ACR, which are located at the end of 22q. Quantitative real-time PCR verified that the deletion of SHANK3 gene in both patients were de novo in origin.</p><p><b>CONCLUSION</b>Two cases of 22q13 deletion syndrome have been diagnosed by SNP array analysis. Deletion of SHANK3 gene may be the major contributor to the clinical manifestations of the patients. SNP array analysis can facilitate discovery of microdeletions, which has played an important role in the diagnosis and genetic counseling for the family.</p>

Predictive Value of CHA2DS2-VASc Score for Contrast-induced Nephropathy After Elective Percutaneous Coronary Intervention in Patients With Coronary Heart Disease / 中国循环杂志

Objectives: To investigate the predictive value of CHA2DS2-VASc score for contrast induced nephropathy (CIN) after percutaneous coronary intervention in patients with coronary heart disease. Methods: A total of 356 patients undergoing elective percutaneous coronary intervention were enrolled in this study. The patients were divided into two groups according to the CHA2DS2-VASc score: CHA2DS2-VASc score ≥ 3 (n=153) and ≤ 2 (n=203). Baseline data, incidence of CIN and major adverse cardiovascular events were analyzed and compared between the two groups. The predictive effect of CHA2DS2-VASc score was analyzed with receiver operating characteristic curve (ROC) and logistic regression analysis. Results: Left ventricular ejection fraction was significantly lower, baseline serum creatinine value was significantly higher, coronary lesions were more complex, contrast agent dosage used was significantly larger and the incidence of CIN was significantly higher in patients of the CHA2DS2-VASc score ≥ 3 group than in patients of CHA2DS2-VASc score ≤ 2 group (all Pvalues<0.05). Multivariate logistic regression analysis showed that CHA2DS2-VASc score≥3 was an independent predictor of CIN (OR=2.152, 95% CI: 1.261-3.987, P=0.032). The area under the curve of ROC of CHA2DS2-VASc score ≥ 3 for predicting CIN was 0.749 (sensitivity 76.9%, specificity 73.0%). Conclusions: CHA2DS2-VASc score could predict the CIN after percutaneous coronary intervention in patients with coronary heart disease, which could help us identify the high-risk patients of CIN and take preventive measures to reduce the incidence of CIN post percutaneous coronary intervention.

Correlation between Movement Distance of Small Intestinal Contents and Survival Time in Female Rats after Feeding / 法医学杂志

OBJECTIVES@#To study the correlation between the movement distance of small intestinal contents and survival time in female SD rat models after one-time satiation, and to evaluate its application value for postmortem interval estimation.@*METHODS@#Adult female SD rats were randomly divided into postprandial groups （1, 2, 3, 4 and 5 h after feeding） and control group. The postprandial groups were fed for 1 h, meanwhile control group was kept fasting. All rats were sacrificed at the given time. The contents in stomach and small intestine were observed, described, compared and photographed, and the movement distance of small intestinal contents was measured. The data of postprandial groups were analysed by one-way analysis of variance.@*RESULTS@#The stomach and duodenum of control group were empty with a little thin and yellow small intestinal liquid. The gastral cavities of 1 h postprandial group were full of undigested food. The evolutionary changes of character, colour and content were observed in the gastric and small intestinal contents of other postprandial groups. The movement distance of intestinal contents increased while the empty part decreased gradually. The differences among the postprandial groups were statistically significant （P<0.05）.@*CONCLUSIONS@#After a 24 h fasting with free drinking and the following 1 h feeding, an ideal animal model can be established successfully on female SD rats, which can provide an experimental basis for postmortem interval estimation based on the changes of small intestinal contents in forensic practice.

Effect of SNORD47 over -expression on proliferation and invasion of U87 -epidermal growth factor receptorvⅢ glioma cells / 中华神经医学杂志

Objective To study the effect of SNORD47 over-expression on proliferation and invasion of U87-epidermal growth factor receptor (EGFR)vⅢ glioma cells. Methods U87-EGFRvⅢ glioma cells at logarithmic phase were assigned into lenti-SNORD47 group, lenti-NC group and blank control group. The recombinant lentiviruses containing lenti-SNORD47 or lenti-NC were transfected into U87-EGFRvⅢ glioma cells of the lenti-SNORD47 group and lenti-NC group, respectively. Forty-eight h after transfection, the SNORD47 expression in the three groups was measured by real time quantitative PCR. Western blotting was used to detect the protein expressions of matrix metalloproteinase (MMP) 2 and MMP9. The proliferation of U87-EGFRvⅢ cells 4, 24, 48, 72 and 96 h after transfection was evaluated by CCK-8 assay and colony formation assay. Transwell assay and wound-healing assay were used to examine the invasion and migration of these cells. Results The SNORD47 expression in the lenti-SNORD47 group was significantly higher than that in the lenti-NC group and control group (P<0.05). The protein expressions of MMP2 and MMP9 in the lenti-SNORD47 group were significantly decreased as compared with those in the lenti-NC group and control group (P<0.05). At 48, 72 and 96 h after transfection, the optical density and number of cloned cells in the lenti-SNORD47 group were significantly decreased as with those in the lenti-NC group and control group (P<0.05). The invasion and migration abilities of U87-EGFRvIII cells in the lenti-SNORD47 group were significantly suppressed as compared with those in the lenti-NC group and control group (P<0.05). Conclusion SNORD47 could inhibit the proliferative and invasive abilities of U87-EGFRvⅢ glioma cells.

Effect of exsomes on SH-SY5Y cells after hypoxic ischemia/reperfusion injury / 中华神经医学杂志

Objective To study the effect of exsomes in SH-SY5Y cells after hypoxic ischemia/reperfusion injury.Methods Human umbilical vein endothelial cell hypoxic ischemia/reperfusion (HUVEC I/R) injury models were established,and the exosomes derived from HUVEC I/R were extracted and identified.SH-SY5Y cell hypoxic ischemia/reperfusion injury models (SH-SY5Y I/R) were established,and cells from SH-SY5Y I/R were divided into control group and exosomes-treatedgroup.The proliferation of SH-SY5Y cells was evaluated by CCK-8 assay 24,48and 72 h after cell inoculation.Transwell assay and wound-healing assay were used to examine the invasion and migration.Hochest33258 staining and Flow cytometry were used to monitor the changes of cell cycle and apoptosis.Expressions of Caspase-3,Bax and Bcl-2 were measured by real-time fluorescence quantificative-PCR and Western blotting.Results As compared with those in the control group,the proliferation abilities of SH-SY5Y cells in exosomes-treated group were significantly promoted (48 h:0.70±0.05 vs.0.94±0.08;72 h:0.83±0.05 vs.1.02±0.06),the cell cycle rate of S phase was significantly increased (14.39％±4.11％ vs.20.54％±3.46％),and G0/G1 phase was statistically decreased (71.26％± 5.24％ vs.66.87％±4.23％,P＜0.05).What's more,cell invasive was significantly promoted (44.00±6.56 vs.70.67±6.11),and relative wound injury area was significantly reduced in the exosomes treated group (0.61±0.07 vs.0.52±0.10);significant differences were noted between the two groups (P＜0.05).The mRNA expressions of Bax and Caspase-3 were significantly decreased and the mRNA expressions of Bcl-2 was significantly increased in the exosomes-treated group as compared with those in the control group (P＜0.05).Conclusion HUVEC I/R-derived exosomes play neuro-protective role in human SH-SY5Y cells after hypoxic I/R injury.

Expressions of CD133 and p53 in colorectal carcinoma and the clinical relvance / 实用医学杂志

Objective To investigate the expressions of CD133 and p53 in colorectal cancer and their clin-ical significances. Methods The expressions of CD133 and p53 in 74 colorectal cancer patients were detected by the immunohistochemistry method. The relationships of CD133 and p53 with the clinicopathological parameters and prognosis were analyzed. Results The positive expression rates of CD133 and p53 in colorectal cancer tissues were 33.8%and 55.4%,respectively. The expression levels of CD133 and p53 were not related to age,sex,tumor location and histological type ,and but were significantly related to the histological differentiation ,TNM stage and distant metastasis(P<0.05,respectively). The Spearman correlation analysis showed that there was no correlation between the levels of CD133 and p53. Conclusions The high expressions of CD133 and p53 in colorectal cancer tissues were closely related to the histological differentiation and TNM stage. CD133 and p53 could be used as important biomarkers for the evaluation of malignant biological behavior,and the diagnosis and prognosis of colorectal cancer.

Phenotypic and genetic analysis of four patients with 13q33-q34 microdeletion / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the correlation between 13q33-q34 microdeletion and clinical phenotype.</p><p><b>METHODS</b>Routine chromosomal banding was performed to analyze the karyotype, while array-based comparative genomic hybridization (aCGH array) and single nucleotide polymorphism array(SNP array) were employed to investigate the genome copy number variations.</p><p><b>RESULTS</b>The karyotype of patient 1 was 46, XY, 9qh+,13qs. Patient 2 showed 46, XX, der (13). Patient 3 showed 46, XX, r(13) (p11.2q32) [43]/45, XX, 13[4]/46, XX, r(13;13) [2]/47, XX, 2r(13;13) [1]. Patient 4 did not undergo chromosome karyotyping analysis. Array analysis showed that four patients have different microdeletions in 13q33-34 region and had common features of 13q33-q34deletion including intellectual disability, facial dysmorphism, microcephaly, hypotonia, low birth weight and genital abnormality.</p><p><b>CONCLUSION</b>The severity of phenotypes showed no correlation with the size of deletion in 13q33-q34. The lower percentage of patients with congenital heart disease suggested a complex pathogenesis of such disease. EFNB2, LIG4 and SOX1 in 13q33-34 region are promising candidates for mental retardation. LIG4 was also a likely candidate for microcephaly.</p>

Clinical and genetic analysis of two unrelated patients with Angelman syndrome and novel UBE3A mutations / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the genetic cause for two familial Angelman syndrome cases and correlation between the clinical phenotypes and their genetic basis.</p><p><b>METHODS</b>Karyotyping analysis and microarray assay were carried out to exclude chromosome anomalies and uniparental disomy. The UBE3A gene was analyzed for potential point mutations, deletions, insertions and splice site mutations. Reverse transcription PCR was used to evaluate splicing mutation of the RNA transcripts.</p><p><b>RESULTS</b>DNA sequencing showed the proband of family 1 has carried a novel maternal UBE3A splice acceptor site mutation, resulting in a guanine-to-cytosine transversion (IVS15-1G>C). Reverse transcription PCR revealed the proband and his mother both carried heterozygous mutant transcripts with loss of 54 and 59 nucleotides in exon 16, respectively. The proband displayed severe mental retardation, ataxia, seizures and inappropriate laughter. The siblings of family 2 has carried a novel maternal missense mutation in exon 16 of the UBE3A gene (c.2540C>T). She also presented with mental retardation, absent speech, mild ataxia and inappropriate laughter.</p><p><b>CONCLUSION</b>The novel IVS15-1G>C and c.2540 C>T mutations of the UBE3A gene probably underlie the AS in the two families. Compared with small-scale mutations, larger fragments mutations can produce more severe phenotypes.</p>