Amentoflavone induces apoptosis and suppresses proliferation of thyroid carcinoma SW579 cells by inhibiting the activation of JAK2-STAT3 pathway / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨阿曼托双黄酮（AF）对甲状腺癌SW579细胞中JAK2-STAT3通路活化及其细胞增殖和凋亡的影响。方法：用0、50、100、150、200 μmol/L的AF处理SW579细胞24、48、72 h，采用CCK-8和Celigo计数、FCM、WB及qPCR法检测AF对SW579细胞的增殖、凋亡、JAK2-STAT3通路活化及其下游调控基因c-Myc、Bcl2、survivin的mRNA及蛋白表达水平的影响。结果：AF处理后，SW579细胞增殖能力显著下降（P<0.05）且呈浓度依赖性，细胞凋亡呈浓度依赖性增多（P<0.05），细胞中JAK2-STAT3通路的活化受到显著抑制（P<0.05），其下游基因c-Myc、Bcl2、survivin的mRNA及蛋白表达均明显下降（均P<0.05）。结论：AF可通过抑制SW579细胞中JAK2-STAT3通路活化及其下游基因的表达而抑制SW579细胞的增殖并促进其凋亡，有望成为治疗甲状腺癌的有效药物。

Enrichment of miR-17-5p enhances the protective effects of EPC-EXs on vascular and skeletal muscle injury in a diabetic hind limb ischemia model

BACKGROUND/AIMS: Diabetes mellitus (DM) is highly susceptible to diabetic hind limb ischemia (DHI). MicroRNA (MiR)-17-5p is downregulated in DM and plays a key role in vascular protection. Endothelial progenitor cell (EPC)-released exosomes (EPC-EXs) contribute to vascular protection and ischemic tissue repair by transferring their contained miRs to target cells. Here, we investigated whether miR-17-5p-enriched EPC-EXs (EPC-EXsmiR-17-5p) had conspicuous effects on protecting vascular and skeletal muscle in DHI in vitro and in vivo. METHODS: EPCs transfected with scrambled control or miR-17-5p mimics were used to generate EPC-EXs and EPC-EXsmiR-17-5p. Db/db mice were subjected to hind limb ischemia. After the surgery, EPC-EXs and EPC-EXsmiR-17-5p were injected into the gastrocnemius muscle of the hind limb once every 7 days for 3 weeks. Blood flow, microvessel density, capillary angiogenesis, gastrocnemius muscle weight, structure integrity, and apoptosis in the hind limb were assessed. Vascular endothelial cells (ECs) and myoblast cells (C2C12 cells) were subjected to hypoxia plus high glucose (HG) and cocultured with EPC-EXs and EPC-EXsmiR-17-5p. A bioinformatics assay was used to analyze the potential target gene of miR-17-5p, the levels of SPRED1, PI3K, phosphorylated Akt, cleaved caspase-9 and cleaved caspase-3 were measured, and a PI3K inhibitor (LY294002) was used for pathway analysis. RESULTS: In the DHI mouse model, miR-17-5p was markedly decreased in hind limb vessels and muscle tissues, and infusion of EPC-EXsmiR-17-5p was more effective than EPC-EXs in increasing miR-17-5p levels, blood flow, microvessel density, and capillary angiogenesis, as well as in promoting muscle weight, force production and structural integrity while reducing apoptosis in gastrocnemius muscle. In Hypoxia plus HG-injured ECs and C2C12 cells, we found that EPC-EXsmiR-17-5p could deliver their carried miR-17-5p into target ECs and C2C12 cells and subsequently downregulate the target protein SPRED1 while increasing the levels of PI3K and phosphorylated Akt. EPC-EXsmiR-17-5p were more effective than EPC-EXs in decreasing apoptosis and necrosis while increasing viability, migration, and tube formation in Hypoxia plus HG-injured ECs and in decreasing apoptosis while increasing viability and myotube formation in C2C12 cells. These effects of EPC-EXsmiR-17-5p could be abolished by a PI3K inhibitor (LY294002). CONCLUSION: Our results suggest that miR-17-5p promotes the beneficial effects of EPC-EXs on DHI by protecting vascular ECs and muscle cell functions.

Impact of oliguria during lung surgery on postoperative acute kidney injury / 北京大学学报(医学版)

OBJECTIVE@#To explore the influence of intraoperative urine volume on postoperative acute kidney injury (AKI) and the independent risk factors of AKI.@*METHODS@#This was a retrospective cohort study recruiting patients who received selective pulmonary resection under general anesthesia in Peking University First Hospital from July, 2017 to June, 2019. The patients were divided into the AKI group and the control group according to whether they developed postoperative AKI or not. Firstly, univariate analysis was used to analyze the relationship between perioperative variables and postoperative AKI. Secondly, receiver operating characteristic (ROC) curve was used to explore the predictive value of intraoperative urine output for postoperative AKI. The nearest four cutoff values [with the interval of 0.1 mL/(kg·h)] at maximum Youden index were used as cutoff values of oliguria. Then univariate analysis was used to explore the relationship between oliguria defined by these four cutoff values and the risk of AKI. And the cutoff value with maximum OR was chosen as the threshold of oliguria in this study. Lastly, the variables with P < 0.10 in the univariate analysis were selected for inclusion in a multivariate Logistic model to analyze the independent predictors of postoperative AKI.@*RESULTS@#A total of 1 393 patients were enrolled in the study. The incidence of postoperative AKI was 2.2%. ROC curve analysis showed that the area under curve (AUC) of intraoperative urine volume used for predicting postoperative AKI was 0.636 (P=0.009), and the cutoff value of oliguria was 0.785 mL/(kg·h) when Youden index was maximum (Youden index =0.234, sensitivity =48.4%, specificity =75.0%). Furthermore, 0.7, 0.8, 0.9, 1.0 mL/(kg·h) and the traditional cutoff value of 0.5 mL/(kg·h) were used to analyze the influence of oliguria on postoperative AKI. Univariate analysis showed that, when 0.8 mL/(kg·h) was selected as the threshold of oliguria, the patients with oliguria had the most significantly increased risk of AKI (AKI group 48.4% vs. control group 25.3%, OR=2.774, 95%CI 1.357－5.671, P=0.004). Multivariate regression analysis showed that intraoperative urine output < 0.8 mL/(kg·h) was one of the independent risk factors of postoperative AKI (OR=2.698, 95%CI 1.260－5.778, P=0.011). The other two were preoperative hemoglobin ≤120.0 g/L (OR=3.605, 95%CI 1.545－8.412, P=0.003) and preoperative estimated glomerular filtration rate < 30 mL/(min·1.73 m2) (OR=11.009, 95%CI 1.813－66.843, P=0.009).@*CONCLUSION@#Oliguria is an independent risk fact or of postoperative AKI after pulmonary resection, and urine volume < 0.8 mL/(kg·h) is a possible screening criterium.

Virus isolation and genotype identification of human respiratory syncytial virus in Guizhou Province, China

ABSTRACT To investigate the genetic variation and molecular epidemiology characteristics of Human Respiratory Syncytial Virus (HRSV) in Guizhou Province, nasopharyngeal aspirates were collected from patients with acute respiratory infection (ARI) in Guizhou Provincial People's Hospital, from December 2017 to March 2018, and inoculated to Hep-2 cells to isolate HRSV. Cells that showed cytopathic effect (CPE) were then confirmed by indirect immunofluorescence assay and reverse transcription. The sequence of the PCR products was determined for HRSV isolates, and the genetic variation was analyzed. Out of 196 nasopharyngeal aspirate samples, HRSV were isolated in 39. The second hypervariable region at the 3' terminal of glycoprotein gene (HVR2) sequence analysis showed that subgroup A was dominant. Seventy-nine percent of the isolates belonged to subgroup A, ON1 genotype, and 21 % belonged to subgroup B, BA9 genotype, which indicates that the dominant HRSV circulating in Guizhou Province was subgroup A, genotype ON1, co-circulating with a less prevalent subgroup B, genotype BA9.

Purification, characterization and antioxidant activities of enzymolysis polysaccharide from grifola frondosa

Our previous study revealed that the antioxidant activity of polysaccharide [coded as FGFP] extracted from Grifola frondosa by enzymolysis treatment was significantly superior than that [coded as GFP] extracted by boiling-water. In this study, one purified polysaccharide fractions [coded as FGFP-11] was obtained from FGFP by purified using DEAE-52 column and Sephacryl S-500HR column. Results indicated that FGFP-11 with MW of 59.82 kDa consisted of mannose, glucose and galactose with a molar ratio of 1.00:16.36:5.25. Fourier Transform Infrared Spectroscopy [FTIR spectrum] of FGFP-11 was similar with that of polysaccharide extracted by boiling-water from Grifola frondosa. These indicated the enzymolysis did not destroy the polysaccharide structure. NMR spectrum showed that FGFP-11 possess alpha-[1-6] glycosidic bond and alpha-[1-3] glycosidic bond configuration. The experiment of Congo red also revealed that FGFP-11 had triple helix stereo-configuration. Moreover, the antioxidant activities of FGFP-11 were improved compared with that of GFP, especially in scavenging of hydroxyl radical and diphenyl picryl hydrazinyl [DPPH] radical

Screening and verification of plasma biomarkers for stable angina pectoris: a differential proteomic analysis / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare and analyze the differentially expressed plasma proteome between patients with stable angina pectoris (SAP) and healthy donors to identify the biomarkers for early diagnosis of SAP.</p><p><b>METHODS</b>Plasma samples from 60 patients with SAP and 60 healthy controls were collected. Twenty samples (100 mL each) randomly selected from each group were pooled and after removing high-abundance proteins from the pooled plasma, two-dimensional gel electrophoresis (2DE) was performed to isolate the total proteins. The protein spots with more than 2 fold changes were selected after 2D analysis using software, and the differentially expressed proteins were identified by MALDI TOF/TOF mass spectrometer. ELISA was performed to detect hemoglobin subunit delta (HBD) levels in 40 randomly selected samples from each group for verification of the results of 2DE.</p><p><b>RESULTS</b>A total of 7 differentially expressed proteins were found in plasma samples from patients with SAP, including 3 up regulated proteins (serum albumin, hemoglobin subunit alpha and hemoglobin subunit delta,) and 4 down?regulated ones (apolipoprotein L1, apolipoprotein C3, apolipoprotein E and complement C4B). ELISA results showed that HBD level was increased in SAP plasma, which was consistent with the results of 2DE.</p><p><b>CONCLUSION</b>Patients with SAP have different plasma protein profiles from those of healthy controls, and HBD may serve as a potential specific biomarker for early diagnosis of SAP.</p>

Exploring Patterns and Trends with Selected Cancer Rates Reported by China National Cancer Registry: Alternative Perspectives and Findings.

Background: National cancer registration reports provide huge potential for identifying patterns and trends of policy, research, prevention and treatment significance. Yet given the range of factors involved in cancer onset, case identification, progression and reporting, pin-pointing this complexity requires systematic thinking and varied strategies of data analysis. Methods: The study extracts data about incidence rates (IRs) and mortality rates (MRs) of lung, stomach, colorectal and liver cancers for 2004, 2006 and 2009 from relevant China National Cancer Registry (CNCR) reports and analyzes the data using line-graphs, ratios and logistic growth modeling. Results: The study shows that: a) all line graphs of age-specific IRs and MRs of the 4 cancers characterized typical S-shape with substantial differences in terms of smoothness, height and proximity; b) MR lines mimicked and located below the corresponding (of the same cancer, population group and year of reporting) IR lines for almost all the age groups except 1 to 2 oldest ones; c) colorectal cancer witnessed the lowest MR/IR ratios on average followed by gastric and lung cancers and all such ratios featured an increasing trend along the age spectrum; d) urban vs. rural ratios in IRs or MRs showed an increasing trend along the age axis for 3 out of the 4 cancers but a typical v-shaped curves for stomach cancer; e) the lines of recent vs. early ratios in cumulative IRs or MRs for urban areas located apparently closer than that for rural areas; f) all the age-specific IRs and MRs fitted very well with logistic growth models (goodness of fit> 0.91) and the integrations and ages when the models reached 5%, 50% or 95% of their highest values yielded interesting features. Conclusion: The study provides useful perspectives for analyzing age-specific IRs and MRs and reveals a number of interesting patterns and trends with cancer counts reported by CNCR.

Strategies for exome sequence data analysis for discovering genes underlying autosomal recessive disorders / 中华医学遗传学杂志

Introduced in 2009, whole-exome sequencing (WES) is a technology in which target capture methods are used to enrich sequences of coding regions of genes from fragmented total genomic DNA, which is followed by high-throughput sequencing of the captured fragments. As reported, WES has been successfully applied for discovering genes underlying several Mendelian diseases, especially autosomal recessive types. In this review, authors have summarized the main computational strategies which have been applied to identify novel autosomal recessive diseases genes using whole-exome data.

A mouse model of myelofibrosis induced by high dose of recombinant human erythropoietin / 中国实验血液学杂志

Tn order to set up a mouse model of myelofibrosis (MF) induced with high dose recombinant human erythropoietin (rhEPO). 60 mice were collected and divided into EPO and control groups, the former was injected with rhEPO and the latter with normal saline intraperitoneally. 5 mice from each group were executed on day 6, 30, 60, 90, 120 and 150 respectively. Their WBC count, Hb level, MCV, RDW and platelet amount were measured by automatic blood cell analyzer; CD34(+) cell ratio in bone marrow were analyzed by flow cytometry; liver and spleen coefficients were measured; pathological changes of liver, spleen, femur were observed by HE staining and reticular fibers staining; cortex thickness, femoral canal diameter and lumbar spine density were determined by computerized tomography (CT). The results indicated that as compared with normal control group in EPO induced group, WBC count was increased slightly in whole period, but without statistic significance (p > 0.05), Hb level and RDW increased at day 6 and 30 significantly (p < 0.05), MCV increased at day 6 significantly (p < 0.05), but platelet amount decreased significantly at all time points (p < 0.05). Most mice in EPO-induced group had hepatomegalia and their liver and spleen coefficient increased significantly at day 60 (p < 0.05), while most mice had splenomegaly and its coefficient was increased significantly at all time-points (p < 0.05). CD43(+) cell ratio of EPO group increased significantly in whole period (p < 0.05). CT scanning displayed femoral cortical thickening, medulla canal narrowing and lumbar spine density increasing at day 150, meanwhile, HE staining and reticular fiber staining showed the fatty degeneration or vacuolization in liver, splenomegaly with megakaryocytic proliferation, femur bone marrow fibrosis and osteosclerosis. It is concluded that the mouse induced by high dose of rhEPO displays the myelofibrosis associated with splenic extramedullary hemopoiesis, and this study is useful to establish a practical MF model, and to explore its pathological mechanism.

Progress in the pathogenesis of acute respiratory distress syndrome in neonate by caesarean section / 国际儿科学杂志

Acute respiratory distress syndrome is one of the common respiratory diseases in neonates. It is more common in neonates by elective cesarean section.The pathogenesis is complicated, while delayed lung fluid clearance is considered playing a role in it.Meconium aspiration, delayed establishment of respiratory reflex, gestational age, contractions before onset, male baby,perinatal asphyxia,maternal diabetes or asthma are thought to be risk factors.

Intra-bone marrow infusion of human cord blood hematopoietic stem/progenitor cells improves hematopoietic reconstitution in NOD-SCID mice / 中国实验血液学杂志

The purpose of this study was to explore the effect of intra-bone marrow infusion (iBMI) of cord blood (CB)-derived hematopoietic stem/progenitor cells (HS/PCs) on human hematopoietic reconstitution in xenotransplanted NOD-SCID mouse model. Aliquots containing 5 x 10(5) CB CD34(+) cells were transplanted into sublethally irradiated NOD-SCID mouse via intravenous infusion (iVI) or iBMI routes. 64 female NOD-SCID mice were divided randomly into 3 groups: iBMI group, iVI group and negative control group. The engraftment levels of human hematopoietic cells at 3, 5 and 8 weeks after xenotransplantation were detected by fluorescence-activated cell sorter (FACS), polymerase chain reaction (PCR), immunohistochemistry and HPC colony formation assay, and long-term hematopoietic reconstitution capacity of HSC was tested by secondary transplantation. The results showed that the percentages of human CD45(+) cells in bone marrow, peripheral blood and spleen of recipient in iBMI group at 8 weeks after xenotransplantation were significantly higher than those in iVI group (p < 0.05). HS/PCs given through both iVI and iBMI methods had the ability of multilineage differentiation, the percentages of CD45(+)CD19(+) cells, CD45(+)CD33(+) cells, CD45(+)CD56(+) cells and CD45(+)CD34(+) cells of recipient bone marrow in iBMI group at 8 weeks after xenotransplantation were significantly higher than those in iVI group (p < 0.05), while other lineages in iBMI group were also higher than that in iVI group (p > 0.05). alpha-satellite-specific fragment of human chromosome 17 could be detected by PCR in liver, spleen, lung, peripheral blood and bone marrow cells of long-term survival recipients in both iVI and iBMI groups. Human CD45 antigen could be detected by immunohistochemical method in spleen, liver and lung of recipients in iBMI group at 8 weeks after xenotransplantation. Total colony count in iBMI group at 8 weeks after xenotransplantation was significantly higher than that in iVI group (p < 0.05). alpha-satellite-specific fragment of human chromosome 17 could be detected in all above organs of both group recipients at 6 weeks after secondary xenotransplantation. It is concluded that iBMI of CB CD34(+) cells improves hematopoietic reconstitution in xenotransplanted NOD-SCID mouse model.

Effect of stromal cell-derived factor-1 and its receptor CXCR4 on liver metastasis of human colon cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of chemokine stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 on liver metastasis of human colon cancer.</p><p><b>METHODS</b>Expression of CXCR4 in different colon cancer cell lines and SDF-1 in different tissues were detected by using Western-blot technique. Effect of SDF-1 and anti-CXCR4 monoclonal antibody (McAb) on proliferation and migration of HT-29 cells were measured using MTT methods. Model mimicking liver metastasis of human colon cancer was established by injecting HT-29 cells intrasplenically into BALB/C nude mice. Mice were randomly divided into AMD3100 treated group and control group. Liver metastatic rate and tumor foci were measured 7 weeks after.</p><p><b>RESULTS</b>HT-29 cells expressed higher level of CXCR4 protein, and liver tissue expressed higher level of SDF-1 protein. Compared with the control, SDF-1 could significantly induced the proliferation and migration of the HT-29 cells, and anti-CXCR4 McAb could inhibited both functions of SDF-1. The liver metastasis rate in the control group was 100%, and it was 40% in the AMD3100 treating group (P < 0.05). The mean liver metastasis number also significantly decreased by AMD3100 (7.8 +/- 2.6 vs 22.4 +/- 8.6, P < 0.05).</p><p><b>CONCLUSIONS</b>SDF-1/CXCR4 biological axis play an important role in liver metastasis of human colon cancer. Arrest of CXCR4 can inhibit liver metastasis of colon cancer through blocking cell proliferation and migration induced by SDF-1.</p>