Objective To investigate the effect of PIAS3 on glucose fluctuation-induced oxidative stress and mito-chondrial dysfunction in rat cardiomyocytes.Methods H9c2 were cultured in vitro,and divided into normal glucose control group(Control),mannitol-induced osmotic pressure control group(MG),constant high glucose group(HG),intermittent hyperglycemia group(IHG),IHG+siRNA NC group,and IHG+PIAS3 siRNA group.Cell proliferation was assessed using CCK-8 assay.LDH release,MDA and GSH levels,as well as SOD activity,were detected using corresponding kits.Mitochondrial membrane potential was evaluated via JC-1 staining combined with flow cytometry.ROS levels in cells and mitochondria were determined using DCFH-DA and MitoSOX staining,re-spectively.Protein expression of PI3K,p-PI3K,AKT,and p-AKT was analyzed by Western blot.Results Com-pared with the control group,intermittent hyperglycemia promoted oxidative stress and mitochondrial dysfunction,significantly upregulated PIAS3 expression(P＜0.001)and downregulated p-PI3K and p-AKT protein levels(P＜0.001).Knockdown of PIAS3 significantly alleviated oxidative stress and mitochondrial dysfunction induced by glucose fluctuations,and increased p-PI3K and p-AKT protein levels(P＜0.001).Conclusions Knockdown of PIAS3 may alleviate glucose fluctuation-induced oxidative stress and mitochondrial dysfunction in ratcardiomyocytes by activating the PI3K/AKT signaling pathway.

Objective To determine the expression of type Ⅲ interferon(IFNL)and IFNL receptor 1(IFNLR1)in the genital tract in rhesus macaques to elucidate the biological characteristics of rhesus macaque IFNL system and promote the use of these monkeys in studies on the pathology,drugs and vaccines for human diseases.Methods IFNL1,IFNL3,and IFNLR1 mRNA levels in normal and simian-human immunodeficiency virus(SHIV)/simian immunodeficiency virus(SIV)-infected genital tract tissues of rhesus macaques were detected by gene-specific reverse transcription-polymerase chain reaction,and IFNLR1 immunoreactivity was examined by confocal microscopy.Results IFNL1 and IFNL3 mRNA levels in the uterus and vagina in normal rhesus macaques were below the measurement threshold,but mRNA levels of IFNLR1 and its variant lacking exon 6 could be readily detected.IFNLR1 immunoreactivity was primarily detected in the covering or glandular epithelia of the uterus and vagina.IFNL1 and IFNL3 mRNA levels were increased in some SHIV/SIV-infected macaques,while mRNA levels of IFNLR1 were unchanged and mRNA levels of IFN-stimulated genes(Mx1,Mx2,and OAS)were significantly increased.Conclusions These data indicate that the type Ⅲ IFN system is expressed in the reproductive tract of rhesus macaques,as in humans,thus providing a scientific basis for the use of rhesus macaques in studies of human infectious diseases,such as AIDS.

Objective To determine the expression of type Ⅲ interferon(IFNL)and IFNL receptor 1(IFNLR1)in the genital tract in rhesus macaques to elucidate the biological characteristics of rhesus macaque IFNL system and promote the use of these monkeys in studies on the pathology,drugs and vaccines for human diseases.Methods IFNL1,IFNL3,and IFNLR1 mRNA levels in normal and simian-human immunodeficiency virus(SHIV)/simian immunodeficiency virus(SIV)-infected genital tract tissues of rhesus macaques were detected by gene-specific reverse transcription-polymerase chain reaction,and IFNLR1 immunoreactivity was examined by confocal microscopy.Results IFNL1 and IFNL3 mRNA levels in the uterus and vagina in normal rhesus macaques were below the measurement threshold,but mRNA levels of IFNLR1 and its variant lacking exon 6 could be readily detected.IFNLR1 immunoreactivity was primarily detected in the covering or glandular epithelia of the uterus and vagina.IFNL1 and IFNL3 mRNA levels were increased in some SHIV/SIV-infected macaques,while mRNA levels of IFNLR1 were unchanged and mRNA levels of IFN-stimulated genes(Mx1,Mx2,and OAS)were significantly increased.Conclusions These data indicate that the type Ⅲ IFN system is expressed in the reproductive tract of rhesus macaques,as in humans,thus providing a scientific basis for the use of rhesus macaques in studies of human infectious diseases,such as AIDS.

Astrocytes are the largest glial population in the mammalian brain. However, we have a minimal understanding of astrocyte development, especially fate specification in different regions of the brain. Through lineage tracing of the progenitors of the third ventricle (3V) wall via in-utero electroporation in the embryonic mouse brain, we show the fate specification and migration pattern of astrocytes derived from radial glia along the 3V wall. Unexpectedly, radial glia located in different regions along the 3V wall of the diencephalon produce distinct cell types: radial glia in the upper region produce astrocytes and those in the lower region produce neurons in the diencephalon. With genetic fate mapping analysis, we reveal that the first population of astrocytes appears along the zona incerta in the diencephalon. Astrogenesis occurs at an early time point in the dorsal region relative to that in the ventral region of the developing diencephalon. With transcriptomic analysis of the region-specific 3V wall and lateral ventricle (LV) wall, we identified cohorts of differentially-expressed genes in the dorsal 3V wall compared to the ventral 3V wall and LV wall that may regulate astrogenesis in the dorsal diencephalon. Together, these results demonstrate that the generation of astrocytes shows a spatiotemporal pattern in the developing mouse diencephalon.
Mice ; Animals ; Astrocytes ; Neuroglia/physiology* ; Diencephalon ; Brain ; Neurons ; Mammals

Objective　To explore the levels of peripheral blood IL-33 and the difference of arteriovenous fistula glucose (Da-jvGlu) after giving Shenqi glucose injection in acute cerebral infarction patients.Methods　The levels of plasma IL-33 and Da-jvGlu were detected in 126 patients of acute cerebral infarction and 10 normal controls.The correlation between IL-33 and cerebral infarction was analyzed.Results　Compared with those in normal control group and the pretherapy groups,the levels of IL-33 in large volume were significantly higher (P<0.05).After giving Shenqi glucose injection,the level of IL-33 in mid volume and small volume were significantly higher between groups (P<0.05).Compared with those in normal control group and the pretherapy groups,the level of Da-jvGlu in large volume was significantly higher (P<0.05).After giving Shenqi glucose injection,there was no significant difference of Da-jvGlu in statics between groups (P>0.05).Conclusion　Shenqi glucose injection can affect the levels of IL-33 to protect nerve cells and stabilize blood glucose concentration.

Objective To investigate how to improve test quality by monitoring and analyzing 15 clinical laboratory quality indicators from the National Health and Family Planning Commission .Methods Data were collected from clinical laboratory department of the First Affiliated Hospital of Kunming Medical University between January 2011 and August 2015.15 quality indicators were analyzed retrospectively , including the error rate of specimen type , the coefficient variation unqualified rate of internal quality control test, the reporting rate of critical value , et al.Results The monitoring results of quality indicators basically satisfied the quality goals , except that the median of turn around time in pre-analytical phase was not established, routine internal quality control was not conducted in some laboratory tests in analytical phase and the reporting rate and reporting timely rate of critical value should be further improved in post -analytical phase .Conclusion Medical laboratory quality system can be continuously improved by means of setting up the quality goals of 15 quality indicators referring to sub-specialty and laboratory tests , as well as automated monitoring, statistics and analysis in LIS.

OBJECTIVETo study the expression of Toll-like receptors (TLRs) mRNA in human trophoblast HTR-8/SVneo cells and the changes in indoleamine 2,3-dioxygenase (IDO) mRNA expression in response to TLR ligand stimulation.

METHODSThe expressions of TLRs and IDO mRNA in human HTR-8/SVneo cells were tested by RT-PCR, and the changes in IDO mRNA levels after exposure to TLR3, TLR4, TLR7/8, and TLR9 ligands were quantitatively analyzed with real-time PCR.

RESULTSIDO and TLR1-10 mRNAs were expressed in HTR-8/SVneo cells. As the cell culture time extended, IDO mRNA expression level tended to increase within 48 h. After stimulation with the TLR ligands, the expression of TLR-3 mRNA was down-regulated while the expression of TLR-4, 7, 8, and 9 mRNA up-regulated. Stimulation of the cells with poly(I:C) lowered the expression of IDO mRNA while IFN-γ increased its expression.

CONCLUSIONSThe expression of IDO mRNA is associated with the nutrition of the maternal-fetal interface. Stimulation with the TLR ligands affects the expression of IDO and TLR mRNA expressions in the cells, which verifies the functional activity of TLRs and suggests a role of IDO in TLR pathway-dependent antiviral immunity.

Cell Line ; Female ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; genetics ; metabolism ; Interferon-gamma ; pharmacology ; Ligands ; Poly I-C ; pharmacology ; RNA, Messenger ; metabolism ; Toll-Like Receptors ; genetics ; metabolism ; Trophoblasts ; cytology ; metabolism

Objective To study the expression of Toll-like receptors (TLRs) mRNA in human trophoblast HTR-8/SVneo cells and the changes in indoleamine 2,3-dioxygenase (IDO) mRNA expression in response to TLR ligand stimulation. Methods The expressions of TLRs and IDO mRNA in human HTR-8/SVneo cells were tested by RT-PCR, and the changes in IDO mRNA levels after exposure to TLR3, TLR4, TLR7/8, and TLR9 ligands were quantitatively analyzed with real-time PCR. Results IDO and TLR1-10 mRNAs were expressed in HTR-8/SVneo cells. As the cell culture time extended, IDO mRNA expression level tended to increase within 48 h. After stimulation with the TLR ligands, the expression of TLR-3 mRNA was down-regulated while the expression of TLR-4, 7, 8, and 9 mRNA up-regulated. Stimulation of the cells with poly(I:C) lowered the expression of IDO mRNA while IFN-γincreased its expression. Conclusions The expression of IDO mRNA is associated with the nutrition of the maternal-fetal interface. Stimulation with the TLR ligands affects the expression of IDO and TLR mRNA expressions in the cells, which verifies the functional activity of TLRs and suggests a role of IDO in TLR pathway-dependent antiviral immunity.

Objective To study the expression of Toll-like receptors (TLRs) mRNA in human trophoblast HTR-8/SVneo cells and the changes in indoleamine 2,3-dioxygenase (IDO) mRNA expression in response to TLR ligand stimulation. Methods The expressions of TLRs and IDO mRNA in human HTR-8/SVneo cells were tested by RT-PCR, and the changes in IDO mRNA levels after exposure to TLR3, TLR4, TLR7/8, and TLR9 ligands were quantitatively analyzed with real-time PCR. Results IDO and TLR1-10 mRNAs were expressed in HTR-8/SVneo cells. As the cell culture time extended, IDO mRNA expression level tended to increase within 48 h. After stimulation with the TLR ligands, the expression of TLR-3 mRNA was down-regulated while the expression of TLR-4, 7, 8, and 9 mRNA up-regulated. Stimulation of the cells with poly(I:C) lowered the expression of IDO mRNA while IFN-γincreased its expression. Conclusions The expression of IDO mRNA is associated with the nutrition of the maternal-fetal interface. Stimulation with the TLR ligands affects the expression of IDO and TLR mRNA expressions in the cells, which verifies the functional activity of TLRs and suggests a role of IDO in TLR pathway-dependent antiviral immunity.

Objective To identify the variation in the Env region of SHIV-XJ02170 during passaging in Chinese origin Rhesus Macaques.Methods Fragments of the SHIV-XJ02170 gp160 and gp120 gene were amplified by PCR and RT-PCR separately from the blood samples of SHIV-XJ02170 infected animals at the peak viral load time point.Purified RT-PCR product was ligated into T easy vector and transformed into JM109 competent cells,18 clones were selected by PCR method and sequenced by ABI 3730DNA sequencers.The gene distances(divergence,diversity)were calculated using DISTANCE.Results In all,the SHTV-XJ02170 gp120 gene evolved forward along the virus passaging.It could be found that viral divergence from the founder strain serially enhanced during in vivo passaging,but in the early phase of each passage,SHIV-XJ02170 gp120 gene evolved toward ancestral state upon transmission to a new host.All of the SHIV-XJ02170 strains had V3 loop central motif(GPGQ)and were predicted to be using CCR5 on the basis of the critical amino acids within V3 loop.Conclusion There was significant increase in the genetic distance during serial passaging,and SHIV-XJ02170 gp120 gene evolved forward along passaging.This could partly explain why the virus infectivity was enhanced during in vivo passaging.