Differential effects of APOE polymorphism in neurotoxicity-responsive astrocytes induced by inflammatory factor / 吉林大学学报(医学版)

Objective:To discuss the differential effects of apolipoprotein E(APOE)gene polymorphism in the neurotoxicity-reactive astrocytes,and to provide the theoretical basis for the study of the pathogenesis of Alzheimer's disease(AD).Methods:The primary cortical astrocytes from the APOE-knockout mice(APOE-/-)were isolated and cultured in vitro,and the purity of the cells was identified by immunofluorescence staining.The human APOE3 and APOE4 recombinant over-expression plasmids were constructed and separately transfected into the primary APOE-/-astrocytes,and the APOE-/-primary cells were regarded as control.Western blotting method was used to detect the expression levels of APOE and glial fibrillary acidic protein(GFAP)proteins in the cells;enzyme-linked immunosorbent assay(ELISA)method was used to detect the APOE level in the cellular culture supernatant.The inflammatory models were prepared with the primary astrocytes transfected with APOE3 and APOE4 and co-stimulated with interleukin-1α(IL-1α),tumor necrosis factor(TNF),and complement C1q.The cells were divided into APOE3+PBS group,APOE4+PBS group,APOE3+IL-1α+TNF+ C1q group,and APOE4+IL-1α+TNF+C1q group.Cell immunofluorescence staining method was used to observe the morphology of the cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of glypican 4(Gpc4),glypican 6(Gpc6),thrombospondin 1(Thbs1),thrombospondin 2(Thbs2),SPARC-like protein 1(Sparcl1)and glial cell line derived neurotrophic factor(GDNF),C3,and S100 calcium binding protein B(S100B)mRNA in the cells in various groups;microsphere phagocytosis assay was used to detect the phagocytic capacities of the cells in various groups;Western blotting was used to detect the protein expression levels of B-cell lymphoma 2(Bcl-2),and cysteinyl aspartate specific protease-3(Caspase-3)proteins in the cells in various groups.Results:Compared with APOE-/-group,the expression levels of APOE and GFAP proteins in the cells and the APOE level in the cellular culture supernatant in transfected APOE3 and transfected APOE4 groups were increased(P<0.01).The fluorescence microscope observation results showed that compared with APOE3+PBS and APOE4+PBS groups,the astrocytic processes in APOE3+IL-1α +TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group became shorter and the cell bodies became larger;compared with APOE3+IL-1α +TNF+Cq1 group,the astrocytic processes in APOE4+IL-1α +TNF+Cq1 group were even shorter.Compared with APOE3+PBS and APOE4+PBS groups,the expression levels of Gpc4,Gpc6,Thbs1,Thbs2,and Sparcl1 mRNA in the cells in APOE3+IL-1α +TNF+Cq1 group and APOE4+IL-1α +TNF+Cq1 group were significantly decreased(P<0.01);compared with APOE3+IL-1α +TNF+Cq1 group,the expression levels of Gpc4,Gpc6,Thbs1,Thbs2,and Sparcl1 mRNA in the cells in APOE4+IL-1α +TNF+Cq1 group were significantly decreased(P<0.05 or P<0.01).Compared with APOE3+PBS and APOE4+PBS groups,the expression levels of GDNF mRNA in the cells in APOE3+IL-1α+TNF+Cq1 group and APOE4+ IL-1α +TNF+Cq1 group were decreased(P<0.01),and the expression levels of C3 and S100B mRNA were increased(P<0.01);compared with APOE3+IL-1α +TNF+Cq1 group,the expression level of GDNF mRNA in the cells in APOE4+IL-1α+TNF+Cq1 group was decreased(P<0.05),and the expression levels of C3 and S100B mRNA were increased(P<0.05).Compared with APOE3+ PBS group and APOE4+PBS group,the numbers of hagocytosis of microspheres in the cells in APOE3+ IL-1α +TNF+Cq1 group and APOE4+IL-1α +TNF+Cq1 group were significantly decreased;compared with APOE3+IL-1α+TNF+Cq1 group,the number of hagocytosis of microspheres in the cells in APOE4+IL-1α+TNF+Cq1 group was significantly decreased.Compared with APOE3+PBS group and APOE4+PBS group,the expression levels of Bcl-2 protein in the cells in APOE3+IL-1α+TNF+ Cq1 group and APOE4+IL-1α +TNF+Cq1 group were decreased(P<0.05 or P<0.01)and the expression levels of Caspase-3 protein were significantly increased(P<0.01);compared with APOE3+ IL-1α+TNF+Cq1 group,the expression level of Bcl-2 protein in the cells in APOE4+IL-1α+TNF+ Cq1 group was decreased(P<0.01),and the expression level of Caspase-3 protein was increased(P<0.05).Conclusion:The APOE4 genotype has a stronger ability to induce the inflammatory factors compared with APOE3;it can lead to a neurotoxicity-reactive astrocyte phenotype,increase the neurotoxicity,affect the astrocyte apoptosis,and aggravate the neuron damage.

A circRNA-miRNA-mRNA network analysis underlying pathogenesis of human heart failure / 老年心脏病学杂志（英文版）

BACKGROUND@#The molecular mechanisms of heart failure (HF) are still poorly understood. Circular RNA (circRNA) has been discovered in the heart in increasing numbers of studies. The goal of this research is to learn more about the potential roles of circRNAs in HF.@*METHODS & RESULTS@#We used RNA sequencing data to identify the characteristics of circRNAs expressed in the heart and discovered that the majority of circRNAs screened were less than 2000 nt. Additionally, chromosomes One and Y had the most and least number of circRNAs, respectively. After excluding duplicate host genes and intergenic circRNAs, a total of 238 differentially expressed circRNAs (DECs) and 203 host genes were discovered. However, only four of the 203 host genes of DECs were examined in HF differentially expressed genes. Another study used Gene Oncology analysis of DECs host genes to elucidate the underlying pathogenesis of HF, and it found that binding and catalytic activity accounted for a large portion of DECs. Immune system, metabolism, and signal transduction pathways were significantly enriched. Furthermore, 1052 potentially regulated miRNAs from the top 40 DECs were collected to build a circRNA-miRNA network, and it was discovered that 470 miRNAs can be regulated by multiple circRNAs, while others are regulated by a single circRNA. In addition, a comparison of the top 10 mRNAs in HF and their targeted miRNAs revealed that DDX3Y and UTY were regulated by the most and least circRNA, respectively.@*CONCLUSION@#These findings demonstrated circRNAs have species and tissue specific expression patterns; while circRNA expression is independent on host genes, the same types of genes in DECs and DEGs worked in HF. Our findings would contribute to a better understanding of the critical roles of circRNAs and lay the groundwork for future studies of HF molecular functions.

Clinical analysis of skeletal malocclusion, axial inclination in patients with short root anomaly of the maxillary central incisors / 口腔疾病防治

Objective @#To investigate the prevalence and distribution of skeletal malocclusion and axial inclination of the maxillary central incisors in short root anomaly (SRA) patients in Kunming city, to provide some reference and guidance for SRA patients′ clinical diagnosis and treatment and prevention in SRA patients. @*Methods@#A total of 1 000 cases were randomly selected from the CBCT database of patients admitted to the author′s hospital from January 2011 to July 2019, and a retrospective analysis was performed. A total of 27 patients with SRA were diagnosed (SRA group).The control group, consisted of 100 randomly selected patients from non-SRA patients. According to the clinical data and cephalometric data, skeletal malocclusion was divided into three subgroups: Class I skeletal malocclusion, Class II skeletal malocclusion and Class Ⅲ skeletal malocclusion. Additionally, the axial inclination of the central incisors was divided into three subgroups: the lingual inclination group, labial inclination group and normal inclination group. The two groups each according to sex, skeletal malocclusion and types of axial inclination of the maxillary central incisors were discussed. @*Results @# The prevalence rate of SRA in the selected population was 2.7%, and the prevalence of SRA in females was 3.67% (21/572) , which was higher than that in males by 1.4% (6/428), and was significantly different between sexes (χ2=4.562, P=0.033). There was a significant difference between SRA patients and control group in terms of skeletal malocclusion (χ2=8.710, P=0.013). Class Ⅲ skeletal malocclusion was the main type of skeletal malocclusion in SRA. There was a significant difference between SRA patients and control group in terms of the axial inclination of the maxillary central incisors (χ2=16.75,P<0.001). Lingual inclination of the maxillary central incisors was the main type of axial inclination of the maxillary central incisors in SRA. @*Conclusion@#There is a certain correlation between class Ⅲ skeletal malocclusion and lingual inclination of the maxillary central incisors and SRA, and the root-crown ratio and root shape of these patients should be evaluated before orthodontics are implemented.

Outcome and prognostic markers of neoadjuvant chemotherapy and cut-off value of Ki-67 in 158 breast cancer patients / 中国肿瘤临床

Objective:To investigate the prognostic value of estrogen receptor (ER), progesterone receptor (PR), and Ki-67 in breast can-cer patients receiving neoadjuvant chemotherapy (NAC) and explore the association of chemotherapy regimens and cycles with the outcome of NAC. Methods:Clinical data of cancer patients receiving NAC were retrospectively analyzed. All the patients were admit-ted in Tianjin Medical University Cancer Institute and Hospital from January 2015 to December 2015. All statistical analyses were per-formed using SPSS version 19.0. The relationship among the outcome of NAC, molecular subtype, expression levels of ER, PR, and Ki-67, and chemotherapy regimens and cycles was investigated. Results:Only five HER-2(+) patients accepted the addition of trastuzum-ab in treatment, and few cases were excluded from the statistical analysis based on the effect of chemotherapy regimens. The effec-tiveness of NAC was positively correlated with the expression of Ki-67 whereas negatively correlated with the expression levels of ER and PR (P<0.05). In patients receiving NAC, the patients with Luminal subtype had worse outcome than those with non-Luminal sub-type (P=0.033). The invalid efficacy of pathologic evaluations of Luminal and non-Luminal NAC were 10.1%and 1.3%, respectively. No significant difference was found in the outcome among patients receiving TE, TEC, or EC-T;however, patients who received more than four cycles of NAC had better outcome than others (P=0.016). The outcome was statistically significant when the cut-off value of Ki-67 was 25%. Conclusion:Ki-67 proliferative index could be used as a prognostic marker to NAC in breast cancer patients. The cut-off value of Ki-67 should be determined on the basis of the data of each cancer patient. The curative effect of NAC was poor, and Luminal pa-tients with chemotherapy were insensitive and could be considered for surgical treatment. Patients who received less than four cycles of NAC had worse outcome than others, and prompt NAC foot treatment could improve the efficiency.

Inhibition of the tubular epithelial-to-mesenchymal transition in vivo and in vitro by the Uremic Clearance Granule () / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of the Uremic Clearance Granule (UCG, ), a Chinese patent medicine, on tubular epithelial-to-mesenchymal transition (EMT) in a unilateral ureteral obstruction (UUO) model in vivo and transforming growth factor (TGF)-β1 induced EMT of HK-2 cells in vitro.</p><p><b>METHODS</b>In vivo study, 50 Sprague Dawley rats were divided into three groups: a sham operation group (n=10), a UUO group (n=20), and a UUO with UCG treatment group (n=20). The UCG was given at a dose of 4.5 g/kg body weight per day by gavage after surgery. In vitro study, HK-2 cells were cultured in 10% fetal bovine serum (FBS), 10% healthy rat serum, 10% FBS and TGF-β1 (10 ng/mL), 10% healthy rat serum and TGF-β1, or 10% rat serum containing the uremic clearance granule and TGF-β1. The expression of the epithelial marker E-cadherin and the mesenchymal markers vimentin and α-smooth muscle actin (α-SMA) in kidney tissues and HK-2 cells were investigated by Western blot analysis and immunofluorescence staining.</p><p><b>RESULTS</b>The rats of the UUO group showed obvious tubulointerstitial fibrosis, compared with the sham operation group rats. Tubulointerstitial fibrosis score was reduced by 17.5%±1.1% at day 7 and by 20.0%±1.2% at day 14 in the UCG-treated group, compared with the UUO group. The UCG could maintained expression of E-cadherin and suppressed expression of vimentin and α-SMA in kidney tissues of UUO rats at days 7 and 14, as determined by Western blot analysis and immunofluorescence staining. Rat serum containing the UCG partially inhibited TGF-β1-induced fibroblast phenotype of HK-2 cells and maintained the epithelial morphology of HK-2 cells in vitro. This occurred partially through a reduction of vimentin expression and an increase of E-cadherin expression.</p><p><b>CONCLUSION</b>These results suggest that the UCG prevents tubular EMT and may be a promising agent for treating tubulointerstitial fibrosis.</p>

Expression of Robo2 gene during the renal development in mice / 解放军医学杂志

Objective To observe the expression of Robo2 gene, and explore its role during the renal development of mice. Methods Real-time quantitative RT-PCR was used to semi-quantitatively measure the expression level of Robo2 mRNA in the developing murine kidney at fetal age of 12.5, 13.5, 14.5, 15.5, 16.5 and 17.5 days, and also 1 day, 1 week, 5 weeks after birth. Immunofluorescence staining was used to examine the expression location of Robo2 protein at different stages of embryonic and postnatal kidney. Results Real-time quantitative RT-PCR analysis revealed that Robo2 was highly expressed in embryonic kidney at fetal age of 12.5, 13.5 and 14.5 days, while the expression level declined quickly thereafter and maintained at very low level after birth. Immunofluorescence staining showed that the expression of Robo2 protein could be primarily detected in metanephric mesenchyme of the developing kidney, but not in the ureteric bud. With the development of embryonic kidney, Robo2 protein was expressed in cell membrane of metanephric mesenchyme, condensed cap mesenchyme surrounding the tip of the ureteric bud, comma-shaped body, S-shaped body and renal capsule, finally expressed in the podocytes. Besides, Robo2 protein was also weakly expressed in part of the proximal tubular epithelial cells. Absence of Robo2 gene resulted in abnormal development of nephron, and broadening of some renal tubules and collecting ducts. Conclusion Robo2 plays an important role in the nephron development in mice by regulating the interaction of metanephric mesenchyme and ureteric bud.

Effective analysis of percutaneous reduction and Kirschner pin fixation for the treatment of intraarticular fractures of the calcaneus in children / 中国骨伤

<p><b>OBJECTIVE</b>To introduce and evaluate the clinical effects of percutaneous reduction and Kirschner pin fixation for the treatment of intraarticular fractures of the calcaneus in children.</p><p><b>METHODS</b>From March 2001 to February 2009,12 patients with intraarticular calcaneal fractures were treated by percutaneous reduction and Kirschner pin fixation (13 feet). There were 8 males and 4 females,ranging in age from 3 to 14,with an average of 8.7 years. According to Essex-Lopresti classification, among 5 feet were tongue fractures and 8 feet were compressed fractures. According to Sanders classification, 9 feet were type II and 4 feet were type III. The Biihler angle and Gissane angle of the calcaneus were obtained before and after operation. All patients were evaluated according to Maryland Foot Score.</p><p><b>RESULTS</b>All the patients were followed up for 16-71 months (means 35.9 months),and all the incisions were healed without complications and infection. The preoperative X-ray film showed that Böhler angle was (19.7+/-5.3) degrees, Gissane angle was (137.3+/-7.5) degrees. The postoperative X-ray film demonstrated that Böhler angle was (32.6+/-3.7) degrees, Gissane angle was (125.4+/-2.9) degrees. There was a significant difference between preoperative and postoperative (P<0.01). The average Maryland score was 96.3+/-2.4 (range, 92 to 100 points).</p><p><b>CONCLUSION</b>Percutaneous reduction and Kirschner pin fixation is an effective minimally invasive way to treat intraarticular fractures of the calcaneus in children, it has many advantages such as minimal invasion, reliable fixation and satisfactory effects.</p>

Internal fixation with AO distal humerus plates for the treatment of distal humeral fractures in elderly osteoporotic patients / 中国骨伤

<p><b>OBJECTIVE</b>To study the clinical effects of AO distal humerus plate (DHP) for the treatment of distal humeral fractures in elderly osteoporotic patients.</p><p><b>METHODS</b>From September 2008 to January 2010, 18 elderly osteoporotic patients with distal humeral fractures were treated with open reduction and internal fixation with DHP. There were 3 males and 15 females, ranging in age from 62 to 83 years (averaged, 71.4 years). According to AO classification, 1 patient was Type A2, 3 patients were Type A3; 2 patients were Type B1, 2 patients were Type B2; 3 patients were Type C1, 4 patients were Type C2, 3 patients were Type C3. The surgical approaches were either bilateral or via olecranon process of ulna. Early mobilization was initiated after surgery. Functional results were evaluated according to the Mayo elbow performance score (MEPS).</p><p><b>RESULTS</b>The average duration of follow-up was 14.6 months (ranged from 11 to 24 months). All the patients had a complete healing of their fractures. There was no infection, hardware failure or loss of reduction after the operations. The average Mayo elbow performance score (MEPS) was (92.2 +/- 9.6) (65 to 100 points), among which the score of pain was (42.5 +/- 5.8) (30 to 45 points), the score of motion range was (19.2 +/- 1.9) (15 to 20 points), the score of stability was (9.2 +/- 1.9) (5 to 10 points), and the score of function was (21.4 +/- 2.9) (15 to 25 points). According to MEPS, 11 patients got excellent results, 5 good and 2 fair.</p><p><b>CONCLUSION</b>Treatment of distal humeral fractures in elderly osteoporotic patients with AO distal humeral plates can obtain immediate stabilization which facilitates early mobilization of the elbow.</p>

Construction of Hexose Transporter-like HXT1 Deletion Mutant in Pichia pastoris / 微生物学通报

Glucose was transported by the large number of hexose transporters in yeast cells. There were 18 hexose transporter genes had been identified in Saccharomyces cerevisiae. However,as an excellent expression system,there was no information of these genes had been reported in Pichia pastoris. Based on high homologous recombination efficiency in yeast,we chose G418 resistance for screening,200 bp were cloned from the up and down sequences of HXT1 ORF respectively,then ligated to the 5′ and 3′ end of G418 resis-tance gene for recombination. After electroporation of GS115 spheroplast and screened through different G418 concentration plates,finally we obtained one HXT1 gene deletion mutant named GS115?HXT1. The growth rate and glucose consumption of this mutant were both lower than the wide type.

Treatment of meniscal injuries of knee joints by arthroscopy / 中国基层医药

Objective To assess the effectiveness of treatment of meniscal injuries of knee joints by arthroscopy.Methods 33 patients 35 joints were followed up and the parts,types and treatment under arthroscopy were analysed.Results 33 patients were followed up from six months to six years,the mean preoperative Lysholm score was 60.5 points,and the mean postoperative one was 86.7 points.Conclusion The advantage of treating meniscal injuries by arthroscopy was the result of correct examination and little wound of arthroscopy operation,and arthroscopic repair or partial menisectomy could effectively restore the function of the injured knee.

Post-traumatic osteolysis of the distal clavicle, pubis and ischium in 7 patients / 中华创伤杂志(英文版)

Post-traumatic osteolysis (PTOL) is a very rare disease occurring after acute trauma or repetitive micro-trauma, which is characterized by persistent pain in the injured site. In this study, we reported 7 patients, in whom osteolysis developed in the distal clavicle, pubis and ischium.