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Objective @# By observing the changes of interleukin-22 ( IL-22) ,signal transduction and transcriptional activator 3 (STAT3) ,fasting blood glucose ( FBG) and fasting insulin ( FINS) of rats under the circumstance of chronic intermittent hypoxia and reoxygenation,to explore the role of IL-22 / STAT3 pathway in insulin resistance in- duced by chronic intermittent hypoxia.@*Methods @#4 SD rats were randomly divided into control group (NC group) and intermittent hypoxia group ( CIH group) ,with 12 rats in each group.NC group was placed in normoxia environment for 12 weeks,while CIH group was first given intermittent hypoxia for 8 weeks and then resumed normoxia feeding until 12 weeks.FBG,FINS,IL-22 and p-STAT3 / STAT3 levels were measured at baseline,week 8 and week 12 in both groups,and insulin resistance index (HOMA-IR) was calculated.The differences between the two groups were compared. @*Results @#① There was no significant difference of the observation indexes between the two groups at baseline (P>0. 05) .At 8 weeks,the levels of FBG,FINS and HOMA-IR in CIH group were higher than those in NC group (P<0. 05) ,and the levels of IL-22 were lower than those in NC group (P <0. 05) .p-STAT3 / STAT3 showed a decreasing trend,but not statistically significant.At 4 weeks of reoxygenation,there were no significant differences in FBG,FINS,HOMA-IR and IL-22 levels between the two groups (P >0. 05 ) .p-STAT3 / STAT3 in CIH group was significantly higher than that in NC group ( P <0. 05 ) . ② Spearman rank correlation analysis showed that HOMA-IR was negatively correlated with IL-22 and p-STAT3 / STAT3 ( all P <0. 05) .@*Conclusion@#Chronic intermittent hypoxia can inhibit the expression of IL-22 / STAT3 signaling pathway,IL-22 / STAT3 signaling pathway may mediate insulin resistance induced by chronic intermittent hypoxia.
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Objective @#To investigate the role of endoplasmic reticulum stress in hepatic insulin resistance induced by intermittent hypoxia in rats.@*Methods @#Twenty-four SD rats were randomly divided into control group ( NC group) and intermittent hypoxia group ( CIH group) .The NC group was placed in a normoxia environment for 12 weeks,and the CIH group was given intermittent hypoxia for 8 weeks,and then returned to normoxia until the 12th week.In both groups,fasting blood glucose (FBG) ,fasting insulin (FINS) ,and liver inositol-requiring enzyme- 1 α(IRE1 α) ,X-box binding protein 1s(XBP1s) ,forkhead box transcription factor O1 (FoxO1) ,activating transcription factor-6(ATF6) ,cAMP-response element binding protein( CREB) ,CREB-regulated transcription coacti- vator-2( CRTC2) ,double-stranded RNA-dependent protein kinase-like ER kinase ( PERK) ,eukaryotic initiation factor 2 α(eIF2 α) ,protein kinase B ( AKT) ,phosphoenolpyruvate carboxykinase ( PEPCK) ,glucose-6-phosphat- ase( G6Pase) mRNA were measured at baseline,week 8,and week 12 .@*Results @#There was no significant differ- ence in each observation index between the two groups at baseline ; at 8 weeks,the levels of FBG,FINS and the mRNA levels of IRE1α , XBP1s,ATF6,PERK,eIF2 α , PEPCK and G6Pase in the CIH group were higher than those in the NC group (P<0. 05) ,while the mRNA levels of CREB,CRTC2 and AKT were lower than those in the NC group (P<0. 05) ; at 12 weeks,there was no significant difference in each observation index between the two groups.Pearson correlation analysis showed(8th week of intermittent hypoxia group) : homeostasis model as- sessment-insulin resistance(HOMA-IR) was positively correlated with FoxO1,CREB,CRTC2 and PERK,eIF2 α mRNA levels (r = 0. 172,0. 595,0. 183,0. 702,0. 608 ; P<0. 05) while it was negatively correlated with IRE1α , XBP1s,ATF6,AKT mRNA levels (r = -0. 422 ,-0. 327 ,-0. 309 ,-0. 399 ; P<0. 05) .@*Conclusion @#Intermittent hypoxia can lead to insulin resistance,and endoplasmic reticulum stress may mediate this effect.
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Objective:To explore the use of internet-based continuous visual recognition task (MemTrax test, MTX) as a rapid screening tool for amnestic mild cognitive impairment (aMCI).Methods:Sixty-four patients with aMCI and 64 individuals with normal cognition as healthy controls were enrolled respectively from Department of Neurology and Health Examination Center of the First Affiliated Hospital of Kunming Medical University from August 2018 to December 2019. Montreal Cognitive Assessment (MoCA) scale and MTX were adopted to assess the cognitive function of all subjects. The total adjusted MoCA scale score, correct rate of MTX, reaction time of MTX and MTX score were obtained and statistically analyzed.Results:The adjusted MoCA scale scores of aMCI patients and healthy controls were 19 (14, 24) and 26 (24, 27; Z=6.795), the correct rate of MTX of aMCI patients and healthy controls were 74% (60%, 80%) and 88% (84%, 94%; Z=8.359), and the MTX score of aMCI patients and healthy controls were 51.11±14.07 and 70.56±14.91 ( t=7.590), respectively, all with statistically significant difference ( P<0.001). Reaction time of MTX of aMCI patients and healthy controls was 1.401 (1.253, 1.590) s and 1.277 (1.163, 1.410) s, respectively ( Z=3.083, P<0.01). After adjustment for age, physical or mental occupation, exercise, hypertension, hyperlipidemia, stroke, sleep time, as well as smoke, the linear regression showed that the aMCI patients had a significant decrease of adjusted MoCA score, correct rate of MTX and MTX score ( P<0.001), and an extension of reaction time of MTX ( P=0.071), compared with the controls. By MTX and MoCA scale assessment, the best cutoff value was 81% for correct rate of MTX and 23 for adjusted MoCA scale score respectively for the prediction of aMCI (with sensitivity of 79.7%, 93.8% respectively, and specificity of 68.8%, 82.8% respectively). The area under the curve (AUC) of correct rate of MTX was 0.93 (95% CI 0.89-0.97, P<0.001), and the AUC of adjusted MoCA score was 0.85 (95% CI 0.78-0.91, P<0.001). There was a statistically significant difference in paired comparison of the two AUCs (χ2=4.620, P<0.05). Conclusion:MTX acts better for the detection of aMCI than MoCA scale, and correct rate of MTX<81% can be considered as the existence of MCI.
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Collaboration of c-KIT mutations with AML1-ETO (AE) has been demonstrated to induce t(8; 21) acute myeloid leukemia (AML). Targeted therapies designed to eliminate AE and c-KIT oncoproteins may facilitate effective treatment of t(8; 21) AML. Homoharringtonine (HHT) features activity against tumor cells harboring c-KIT mutations, whereas oridonin can induce t(8; 21) AML cell apoptosis and AE cleavage. Therefore, studies should explore the efficacy of combination therapy with oridonin and HHT in t(8; 21) AML. In this study, we investigated the synergistic effects and mechanism of oridonin combined with HHT in t(8; 21) AML cell line and mouse model. The two drugs synergistically inhibited cell viability and induced significant mitochondrial membrane potential loss and apoptosis. Oridonin and HHT induced significant downregulation of c-KIT and its downstream signaling pathways and promoted AE cleavage. HHT increased intracellular oridonin concentration by modulating the expressions of MRP1 and MDR1, thus enhancing the effects of oridonin. The combination of oridonin and HHT prolonged t(8; 21) leukemia mouse survival. In conclusion, oridonin and HHTexert synergistic effects against t(8; 21) leukemia in vivo and in vitro, thereby indicating that their combination may be an effective therapy for t(8; 21) leukemia.
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AIM:To study the effects of methylene blue (MB) on myocardial ischemia/reperfusion (I/R)-induced mitochondrial injury in isolated rat hearts.METHODS:Spragure-Dawley (SD) rats were divided into 3 groups randomly (n=6): control group, I/R model group and MB treatment group (IR+MB group).The isolated rat hearts were prepared and set up to Langendorff perfusion.The rats in I/R+MB group received MB (2 mg/kg) by intraperitoneal injection 2 h before operation.The hearts in control group were perfused with K-H solution for 110 min consecutively.The hearts in I/R group and I/R+MB group were in equilibrium for 20 min, following by 45 min of global ischemia, and then reperfused for 60 min.The heart rate (HR), left ventricular developed pressure (LVDP), left ventricular pressure maximum change rate (±dp/dtmax) and left ventricular end-diastolic pressure (LVEDP) were recorded.The perfusate was collected to determine the activity of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH).The contents of reactive oxygen species (ROS), malondialdehyde (MDA) and adenosine triphosphate (ATP), and the activity of superoxide dismutase (SOD) in the myocardial tissues were all determined.Histopathological examination of left ventricle was performed.The mitochondria from the heart tissues was isolated and the mitochondrial swelling and mitochondrial membrane potential (MMP) were analyzed.RESULTS:Compared with control group, the hearts in I/R group showed poorer function, higher CK-MB and LDH levels in the perfusate, increased ROS and MDA contents, higher SOD activity and less ATP content in the heart tissues (P<0.05).Furthermore, the mitochondrial swelling level increased and MMP reduced in I/R group (P<0.05).Compared to I/R group, MB improved heart function and reduced the release of CK-MB and LDH (P<0.05).MB also decreased ROS and MDA contents, and increased the activity of SOD and the content of ATP (P<0.05).In addition, MB alleviated mitochondrial swelling and restored the reduced MMP (P<0.05).CONCLUSION: MB protects the isolated rat hearts from I/R-induced injury by attenuating the damage of mitochondria.
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Objective To investigate the method of effectively density gradient centrifugation combined with swim-up improves sperm nuclear integrity and determine whether the sperm chromatin dispersion test of sperm DNA fragmentation in raw or DGC-swim-up treated semen can influence the outcome of IVF. Method The DNA integrity of spermatozoa from 120 patients underwent IVF were analyzed by SCD before and after DGC and swim-up. The predictive value of the SDFI for IVF outcomes were assessed in a cohort of 100 patients who were underwent new embryo transfer. Result In male infertility group,DGC combined with swim-up decreased the SDFI from 22.75(14.44,30.25)to 11.50(5.60,22.79),while the control group decreased the SDFI from 20.86(15.00,26.81) to 7.50(3.63,15.44),respectively(P<0.05);SDFI after optimization in clinical pregnancy group was significantly lower than that of non-pregnant group. The area under the receiver operating characteristic curve was 0.667. The patients with low sperm DFI had a higher implantation rate and pregnancy rate compared with patients with high sperm DFI. Conclusions DGC and swim-up treated Sperm DNA fragmentation can predict the outcome of IVF. The effect of semen optimization on the rate of sperm DNA fragmentation is limited,once exceed,pregnancy rate and birth rate are decreased although fertilization is normal.
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Objective To investigate the method of effectively density gradient centrifugation combined with swim-up improves sperm nuclear integrity and determine whether the sperm chromatin dispersion test of sperm DNA fragmentation in raw or DGC-swim-up treated semen can influence the outcome of IVF. Method The DNA integrity of spermatozoa from 120 patients underwent IVF were analyzed by SCD before and after DGC and swim-up. The predictive value of the SDFI for IVF outcomes were assessed in a cohort of 100 patients who were underwent new embryo transfer. Result In male infertility group,DGC combined with swim-up decreased the SDFI from 22.75(14.44,30.25)to 11.50(5.60,22.79),while the control group decreased the SDFI from 20.86(15.00,26.81) to 7.50(3.63,15.44),respectively(P<0.05);SDFI after optimization in clinical pregnancy group was significantly lower than that of non-pregnant group. The area under the receiver operating characteristic curve was 0.667. The patients with low sperm DFI had a higher implantation rate and pregnancy rate compared with patients with high sperm DFI. Conclusions DGC and swim-up treated Sperm DNA fragmentation can predict the outcome of IVF. The effect of semen optimization on the rate of sperm DNA fragmentation is limited,once exceed,pregnancy rate and birth rate are decreased although fertilization is normal.
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AIM: To investigate the effects of caveolin-1 (Cav-1) scaffolding domain peptide, cavtratin, on lipopolysaccharide (LPS)-induced mouse acute lung injury and heme oxygenase-1 (HO-1) activity.METHODS: Adult male BALB/c mice were randomly divided into 6 groups (n=8 to 10): control, Antennapedia internalization sequence (AP), LPS, LPS+hemin, LPS+ hemin+cavtratin and LPS+hemin+cavtratin+zinc protoporphyrin IX (ZnPP) groups.After LPS administration for 24 h, the lung pathological changes, the wet/dry weight (W/D) ratio of lung tissues, total cell number in bronchoalveolar lavage fluid and serum lactate dehydrogenase activity were measured.The co-localization of HO-1 and Cav-1 was displayed by immunofluorescence, and the HO-1 activity were detected.The mRNA expression of pro-inflammatory cytokines IL-1β, IL-6, TNF-α, MCP-1 and iNOS was detected by real-time PCR.RESULTS: The mice in LPS+hemin+cavtratin group had the decreased interaction between HO-1 and Cav-1, and the increased HO-1 activity compare with LPS group (P<0.05).Compared with LPS group, the pulmonary damage was attenuated in LPS+hemin+cavtratin group, and the injury indexes, including W/D ratio, total cell number in bronchoalveolar lavage fluid and lactate dehydrogenase activity in the serum, and the mRNA expression of inflammatory cytokines all decreased (P<0.05).HO-1 activity inhibitor ZnPP abolished the above protective effect of cavtratin on the lung tissues with LPS-induced acute lung injury.CONCLUSION: Cavtratin has beneficial effects on the lung with LPS-induced acute injury by restoring the HO-1 activity.
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Objective To evaluate the value of whole genome amplification (WGA) combined with array comparative genomic hybridization (aCGH) in prenatal diagnosis. Methods Array CGH were performed by the DNA of 18 prenatal specimens , which were amplified by WGA because of the low DNA yield. Result 3 of the 18 fetuses were 45, X0 and 9 of 15 fetuses with normal aCGH results showed healthy outcome. Conclusion It’ s feasible for prenatal diagnosis using WGA combined with aCGH which not only can shorten the reporting time but also keep the sensitivity and accuracy of detection.
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<p><b>OBJECTIVE</b>To assess the value of array-based comparative genomic hybridization (array-CGH) for analyzing tissues derived from spontaneous abortion and stillbirth.</p><p><b>METHODS</b>Agilent Human Genome CGH Microarray 4×44 K chip and Affymetrix Cytoscan 750 K Array were utilized to detect genome-wide copy number variations (CNV) in 43 fetuses with spontaneous abortion and stillbirth. All identified CNV were analyzed with references from Database of Genomic variants (DGV), database of DECIPHER, ISCA and OMIM, as well as comprehensive literature review to determine whether the identified CNVs were pathogenic. Parental DNA of two cases was also analyzed with the same arrays for pathogenic or unknown significant CNVs.</p><p><b>RESULTS</b>All of the 43 specimens were successfully analyzed. Clinically significant chromosomal aberrations were identified in 32 (74.4%) of the samples, which included 26 aneuploidies and 10 pathogenic CNV.</p><p><b>CONCLUSION</b>Array-CGH is a fast and effective method for analyzing tissues derived from spontaneous abortions and stillbirths which may be difficult to culture for karyotype analysis.</p>
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Adulte , Femelle , Humains , Grossesse , Jeune adulte , Avortement spontané , Diagnostic , Génétique , Aberrations des chromosomes , Hybridation génomique comparative , Méthodes , Variations de nombre de copies de segment d'ADN , Foetus , Chimie , Caryotypage , Complications de la grossesse , Diagnostic , Génétique , Mortinatalité , GénétiqueRésumé
<p><b>OBJECTIVE</b>To reprogram the 1q21.1 microdeletion pluripotent stem cells in order to establish an ideal model for further studying its pathogenesis.</p><p><b>METHODS</b>Human amniotic fluid-derived cells induced pluripotent stem cells (hAF-iPSCs) were induced from amniotic fluid cells harboring the 1q21.1 microdeletion by retroviral vectors encoding Oct4, Sox2, c-Myc and Klf4. Characteristics of the 1q21.1 microdeletion hAF-iPSCs were determined, which included in vitro pluripotency, karyotype, microarray analysis, the capacity of differentiation in vivo and in vitro, etc.</p><p><b>RESULTS</b>hAF-iPSCs derived from amniotic fluid cells harboring the 1q21.1 microdeletion have maintained self renewal, with expression of pluripotency marker genes detectable at mRNA level. Stem cell surface antigens were tested by immunocytochemistry. The 1q21.1 microdeletion hAF-iPSCs showed a normal karyotype after long-term culturing in vitro, and harbored the same microdeletion as confirmed by microarray analysis. The cells have maintained their differentiation capacity both in vivo and in vitro.</p><p><b>CONCLUSION</b>The hAF-iPSCs harboring the 1q21.1 microdeletion have all the characteristics of normal pluripotent stem cells, and can be used for directed differentiation into specific cells, which may provide an ideal model for studying the pathogenesis of 1q21.1 microdeletion in vitro.</p>
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Adulte , Animaux , Femelle , Humains , Mâle , Souris , Grossesse , Jeune adulte , Malformations multiples , Embryologie , Génétique , Liquide amniotique , Biologie cellulaire , Différenciation cellulaire , Délétion de segment de chromosome , Chromosomes humains , Génétique , Chromosomes humains de la paire 1 , Génétique , Maladies foetales , Génétique , Délétion de gène , Cellules souches pluripotentes induites , Biologie cellulaire , Mégalencéphalie , Embryologie , Génétique , Souris SCID , Modèles biologiquesRésumé
BACKGROUND:Kunming mouse embryonic fibroblasts are the most common feeder layers at present, and there are rare reports addressing C57BL/6 mouse embryonic fibroblasts as feeder layers. OBJECTIVE:To separate and culture C57BL/6 mouse embryonic fibroblasts in vitro, and produce feeder layers to enlarge the resources of mouse embryonic fibroblasts. METHODS:C57BL/6 mouse embryonic fibroblasts were isolated and cultured by trypsin digestion method in vitro. The biological characteristics and growth rule of the fibroblasts were investigated, then the feeder layers for the cel culture were produced. The growth of cel colonies on the prepared feeder layer was tested. RESULTS AND CONCLUSION:C57BL/6 mouse embryonic fibroblasts grew wel with a large amount, by trypsin digestion method at different concentrations. There was no significance in the survival rate after cryopreservation for 1 week, 2 weeks, 1 month, 3 months and 6 months. The cel s were proliferative from the second to fifth passage and declined sharply after the sixth passage. The planted mouse embryonic fibroblasts feeder layers had a high activity within 3 days, but got a sharp decline after 4 days. So it is best to use C57BL/6 mouse embryonic fibroblast feeder layers within 3 days after they’re inactivated. C57BL/6 mouse embryonic fibroblast feeder layer can support embryonic stem cel s and induce pluripotent stem cel s to grow as Kunming mouse embryonic fibroblasts.
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Objective To probe into the efficacy and nursing intervention method for simple rib fracture treated by Skeleton Muscle Disease Healing Membrane(SMDHM)combined with TDP irradiation. Methods 120 patients with rib fracture were randomly divided into treatment group and control group. For the treatment group,the patients stuck SMDHM to the place of rib fracture,and then were treated by TDP irradiation. The control group was treated by the conventional method of fixation by rib bandage. Results The fracture healing time of treatment group was short, and antaigic effect was prominent. While, the healing time of control group was long and antaigie effect was poor. For the efficacy observation of the two groups,the statistical difference was extremely significant(P< 0.05). Conclusion The efficacy of SMDHM combined with TDP irradiation for fib fracture is distinctly superior to conventional method. This new treatment is simple,practical and economical, and is easily accepted by patients. Therefore it's worth extensively popularizing in the clinic application.
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Objective To establish effective method for large-scale purification of islet cells from pig pan-cress. Methods Pig pancreas tissue was digested with collagenase P followed by purification in a HCA-Fi-coil dis continuous gradient using Cobe2991 cell separator. After isolation, the islet cell yield and purity were evaluated with light microscope with DTZ staining, and the islet function assessed by insulin release as-say in vitro. Results The number of the islets coll ected from each pancreas averaged (275 000±20 895)islet equivalents (IEQ) before purification, and (230 350±26 679) IEQ after the purification with discon-tinuous gradient centrifugation. From each gram of the pancreatic tissue, (2710±229) IEQ were obtained with an average purity of (50.2±1.95) %. The purified islets responded well to high-concentration (16.7 mmol/L) glucose stimulation with a 4. 74-fold increase of insulin secretion over the basal level (3.3 mmol/L, P <0.001). Conclusion The established method can be applicable for large-scale purifi-cation of fully functional islet cells from pig pancreas.
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@#Objective To observe the effect of early rehabilitation training combined with continuous passive motion (CPM) on knee joint function after total knee arthroplasty (TKA).Methods23 patients (28 knees) post TKA were randomly divided into the treatment group (15 knees) and control group (13 knees). The patients of the treatment group were treated with early rehabilitation training combined with CPM, and those of the control group were treated with CPM only.ResultsThe scores of knee grade and function grade of the treatment group were not different from those of the control group before treatment ( P>0.05), but two weeks later, there was a significnat difference between two groups ( P<0.01).ConclusionThe early rehabilitation training combined with CPM can improve recovery of function of knee joint after TKA.