Abstract Objective: Studies focusing on bone and joint infections (BJIs) in young infants are rare. Some cases of BJI are accompanied by sepsis. This study aimed to identify the clinical and bacteriological features of sepsis in neonates and young infants with BJIs. Methods: Neonates and infants younger than 3 months diagnosed with BJI in the present institution from 2014 to 2021 were retrospectively reviewed. Patient characteristics, clinical data, and outcomes were documented and compared between those with and without sepsis. Results: Twenty-five patients with a mean age of 34.8 days were included. Nine BJI cases had concomitant sepsis (group A), and 16 had BJI without sepsis (group B). Within group A, staphylococcus aureus was the major pathogenic germ (5 cases, of which 4 were of the methicillin-resistant staphylococcus aureus (MRSA) type). There was no statistical difference in male-to-female ratio, age, history of hospitalization, anemia, birth asphyxia, peripheral leukocyte counts, C-reactive protein on admission, and sequelae between groups. Univariate analyses indicated a significant difference in the incidence of septic arthritis (SA) combined with osteomyelitis (OM) (88.9% vs 37.5%), congenital deformities (44.4% vs 0%), and mean duration of symptoms (2.83 days vs 9.21 days) in comparisons between groups A and B. Conclusion: Staphylococcus aureus is the main pathogenic bacteria in BJI cases complicated with sepsis in neonates and young infants. Among infants younger than 3 months diagnosed with BJI, those with concurrent SA and OM, MRSA infection, or congenital deformities are more likely to develop sepsis. Objective BJIs (BJIs rare Methods 201 202 reviewed characteristics data Results Twentyfive Twenty five 348 34 8 34. included , A) 1 B . B) 5 ( methicillinresistant methicillin resistant (MRSA type. type type) maletofemale male female ratio hospitalization anemia asphyxia counts Creactive C reactive admission (SA OM (OM 88.9% 889 88 9 (88.9 37.5%, 375 37.5% 37 37.5%) 44.4% 444 44 (44.4 0%, 0 0% 0%) 2.83 283 2 83 (2.8 921 21 9.2 Conclusion infection 20 88.9 (88. 37.5 44.4 (44. 2.8 28 (2. 92 9. 88. (88 37. 44. (44 2. (2 (8 (4

Abstract Background In a recent genome-wide association study, novel genetic variations of WNT9A were reported to be involved in the etiopathogenesis of thumb osteoarthritis (TOA) in Caucasians. Our purposes were to replicate the association of WNT9A with the development of TOA in the Chinese population and to further unveil the functional role of the risk variants. Methods SNP rs11588850 of WNT9A were genotyped in 953 TOA patients and 1124 healthy controls. The differences of genotype and allele distributions between the patients and healthy controls were evaluated using the Chi-square test. Luciferase Reporter Assay was performed to investigate the influence of variant on the gene expression. Results There was significantly lower frequency of genotype AA in TOA patients than in the controls 74.9% vs. 81.9%, p < 0.001). The frequency of allele A was remarkably lower in the patients than in the controls (86.3% vs. 90.5%, p < 0.001), with an odds ratio of 0.66 (95% CI = 0.54–0.80). Luciferase Reporter Assay showed that the construct containing mutant allele G of rs11588850 displayed 29.1% higher enhancer activity than the wild allele A construct (p < 0.05). Conclusions Allele G of rs11588850 was associated with the increased risk of TOA possibly via up-regulation of WNT9A expression. Further functional analysis into the regulatory role of rs11588850 in WNT9A expression can shed new light on the genetic architecture of TOA. Key Points Genetic variants of WNT9A were associated with the incidence and severity of TOA. Allele G of rs11588850 was associated with an increased transcriptional activity of WNT9A promoter. Allele G of rs11588850 may add to the risk of TOA possibly via up-regulation of WNT9A expression. Further functional analysis into the regulatory role of rs11588850 in WNT9A expression can shed new light on the genetic architecture of TOA. genomewide genome wide study WNTA WNT (TOA Caucasians rs rs1158885 95 112 Chisquare Chi square test 749 74 9 74.9 vs 819 81 81.9% 0.001. 0001 0.001 . 0 001 0.001) 86.3% 863 86 3 (86.3 905 90 5 90.5% 0.001, , 066 66 0.6 95% (95 0.54–0.80. 054080 0.54–0.80 54 80 0.54–0.80) 291 29 1 29.1 0.05. 005 0.05 05 0.05) upregulation up regulation promoter rs115888 11 7 74. 8 81.9 000 0.00 00 86.3 (86. 90.5 06 6 0. (9 05408 0.54–0.8 2 29. 0.0 rs11588 81. 86. (86 90. ( 0540 0.54–0. rs1158 (8 054 0.54–0 rs115 0.54– rs11 0.54 rs1 0.5

Abstract Background: Acid-sensing ion channel 1a (ASIC1a) plays a critical role in physiological and pathological processes. To further elucidate the biological functions of ASICs and their relationships with disease occurrence and development, it is advantageous to investigate and develop additional regulatory factors for ASICs. Methods: In this study, cation exchange chromatography was used to separate seven chromatographic components from Naja naja atra venom. Capillary electrophoresis was employed to detect that Ⅶ peak component containing a main protein Ⅶ-2, which could bind to ASIC1a. The analgesic effects of Ⅶ-2 protein were determined using hot plate methods, and ASIC1a expression in spinal cord tissue from rats with inflammatory pain was detected using western blot. Results: The purified Ⅶ-2 protein named Naja naja atra venom-Ⅶ-2 (NNAV-Ⅶ-2) was obtained by Sephadex G-50 gel filtration, which exhibited a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a molecular weight of 6.7 kD. Remarkably, the NNAV-Ⅶ-2 protein demonstrated a significant analgesic effect and downregulated ASIC1a expression in the spinal cord tissue of rats with inflammatory pain. Conclusions: The analgesic mechanism of the NNAV-Ⅶ-2 protein may be associated with its binding to ASIC1a, consequently downregulating ASIC1a expression in neural tissues. Background Acidsensing Acid sensing ASICa ASIC (ASIC1a processes development Methods study venom Ⅶ2, Ⅶ2 2, 2 Ⅶ- methods blot Results venomⅦ2 venom-Ⅶ- NNAVⅦ2 NNAV (NNAV-Ⅶ-2 G50 G 50 G-5 filtration sulfatepolyacrylamide sulfate polyacrylamide 67 6 7 6. kD Remarkably NNAV-Ⅶ- Conclusions tissues venomⅦ venom-Ⅶ NNAVⅦ (NNAV-Ⅶ- G5 5 G- NNAV-Ⅶ venom- (NNAV-Ⅶ NNAV- (NNAV- (NNAV

Chimeric antigen receptor (CAR) T-cell therapy is a revolutionary immunotherapeutic strategy that has shown efficacy in hematological malignancies. However, its application in solid tumors, particularly gastrointestinal cancers, faces significant challenges. These include the selection of target antigens, the complexity of the tumor microenvironment, and safety and toxicity concerns. This review provides a current overview of CAR-T therapy in various gastrointestinal cancers, such as esophageal, gastric, colorectal, pancreatic, and liver cancers. It discusses the limitations and future directions of CAR-T therapy in this context. This review highlights innovative strategies, including novel target antigens, multispecific CAR-T cells, armored CAR-T cells, and the development of universal CAR-T cells. These insights aim to inform ongoing research and foster advancements in CAR-T therapy for gastrointestinal cancers. CAR (CAR Tcell T cell malignancies However tumors cancers challenges antigens microenvironment concerns CART esophageal gastric colorectal pancreatic context strategies cells

Abstract Objective: this study aimed to evaluate whether low-salt low-protein diet (LPD) supplemented with 10 g of inulin could lower serum toxin levels in patients with chronic kidney disease (CKD), thereby providing evidence for adjusting dietary prescriptions of inhospital patients and outpatient nutrition consultants. Methods: we randomized 54 patients with CKD into two groups. Dietary protein intake compliance was evaluated using a 3-day dietary diary and 24-h urine nitrogen levels. The primary outcomes were indoxyl sulfate (IS) and p-cresyl sulfate (PCS), and secondary outcomes included inflammation marker levels, nutritional status, and renal function. We assessed 89 patients for eligibility, and a total of 45 patients completed the study, including 23 and 22 in the inulin-added and control groups, respectively. Results: PCS values decreased in both groups after intervention: inulin-added group, ∆PCS -1.33 (-4.88, -0.63) µg/mL vs. LPD group, -4.7 (-3.78, 3.69) µg/mL (p = 0.058). PCS values reduced from 7.52 to 4.02 µg/mL (p < 0.001) in the inulin-added group (p < 0.001). Moreover, IS decreased from 3.42 (2.53, 6.01) µg/mL to 2.83 (1.67, 4.74) µg/mL after adding inulin; ∆IS was -0.64 (-1.48, 0.00) µg/mL, and a significant difference was observed compared with the control group (p = 0.004). The inflammation index decreased after intervention. Conclusion: dietary fiber supplementation may reduce serum IS and PCS levels and modulate their inflammatory status in predialysis CKD patients.

Resumen Objetivo: este ensayo aleatorizado doble ciego comparó el efecto de una dieta baja en proteínas (LPD) con o sin suplementos orales de 10 g de inulina en los niveles de PBUT en pacientes con ERC en prediálisis durante 12 semanas. Métodos: clasificamos aleatoriamente a 54 pacientes con ERC en dos grupos. El cumplimiento de la ingesta dietética de proteínas se evaluó utilizando un diario dietético de 3 días y nitrógeno en orina de 24 horas. Los resultados primarios fueron IS y PCS y los resultados secundarios incluyeron niveles de marcadores de inflamación, estado nutricional y función renal. Evaluamos la elegibilidad de 89 pacientes y 45 completaron la intervención, incluidos 23 y 22 en los grupos de inulina añadida y de control, respectivamente. Resultados: el sodio urinario promedio de 24 horas fue de 86 mmol/día y la ingesta promedio de proteínas fue de ~0,7 g/kg/día. Los valores de PCS exhibieron una tendencia decreciente en ambos grupos después de la intervención: grupo con inulina añadida, ∆PCS -1.33 (-4.88, -0.63) µg/mL vs. grupo LPD, -4.7 (-3.78, 3.69) µg/mL) (p =0,058). Los valores de PCS se redujeron de 7,52 a 4,02 µg/mL (p < 0,001) con inulina (p < 0,001). Además, IS disminuyó de 3,42 (2,53, 6,01) µg/mL a 2,83 (1,67, 4,74) µg/mL después de agregar inulina; El ∆IS fue -0,64 (-1,48; 0,00) µg/mL y se observó una diferencia significativa en comparación con el grupo control (p =0,004). Conclusión: la suplementación con fibra dietética puede reducir las toxinas de unión a proteínas séricas en pacientes con ERC en prediálisis y modular su estado inflamatorio.

ABSTRACT Objective: To explore the factors affecting short-term prognosis of circulatory failure patients undergoing venoarterial extracorporeal membrane oxygenation (VA-ECMO) treatment. Methods: A total of 136 patients undergoing VA-ECMO were enrolled in this study and subsequently divided into the death group (n=35) and the survival group (n=101) based on whether death occurred during hospitalisation. Extracorporeal membrane oxygenation (ECMO) running time, length of intensive care unit stay, length of hospital stay, costs, and ECMO complications were then compared between the two groups. Results: The average age of all patients undergoing ECMO was 47.64±16.78 years (53.2±16.20 years in the death group and 45.713±16.62 years in the survival group) (P=0.022). Patients in the survival group exhibited a clear downward trend in lactic acid value following ECMO treatment compared to those in the death group. Total hospitalisation stay was longer in the survival group (35 days) than in the death group (15.5 days) (P<0.001). In the analysis of ECMO complications, the incidence of neurological complications, renal failure, limb complications, and infection were higher in the death group than in the survival group (P<0.05 for all). Specifically, as a risk factor for patient survival and discharge, the occurrence of infection will lead to increased hospitalisation stays and costs (P<0.05 for both). Conclusion: Complications such as kidney failure and infection are associated with in-hospital death, and ECMO-related complications should be actively prevented to improve the survival rate of VA-ECMO treatment. Objective shortterm short term VAECMO VA (VA-ECMO Methods 13 n=35 n35 n 35 (n=35 n=101 n101 101 (n=101 (ECMO time groups Results 47641678 47 64 16 78 47.64±16.7 53.2±16.20 5321620 53 2 20 (53.2±16.2 457131662 45 713 62 45.713±16.6 P=0.022. P0022 P P=0.022 . 0 022 (P=0.022) (3 days 15.5 155 15 5 (15. P<0.001. P0001 P<0.001 001 (P<0.001) P<0.05 P005 05 (P<0.0 all. all) Specifically discharge both. both both) Conclusion inhospital ECMOrelated related 1 n=3 n3 3 (n=3 n=10 n10 10 (n=10 4764167 4 6 7 47.64±16. 53.2±16.2 532162 (53.2±16. 45713166 71 45.713±16. P002 P=0.02 02 (P=0.022 ( 15. (15 P000 P<0.00 00 (P<0.001 P<0.0 P00 (P<0. n= (n= n=1 n1 (n=1 476416 47.64±16 53.2±16. 53216 (53.2±16 4571316 45.713±16 P=0.0 (P=0.02 (1 (P<0.00 P<0. P0 (P<0 (n 47641 47.64±1 53.2±16 5321 (53.2±1 457131 45.713±1 P=0. (P=0.0 P<0 (P< 4764 47.64± 53.2±1 532 (53.2± 45713 45.713± P=0 (P=0. P< (P 476 47.64 53.2± (53.2 4571 45.713 P= (P=0 47.6 53.2 (53. 457 45.71 (P= 47. 53. (53 45.7 (5 45.

Abstract Under the influence of the COVID-19, people's awareness of physical health and immunity has increased significantly. Chitooligosaccharide is an oligomer of β-(1, 4)-linked D -glucosamine, furthermore, is one of the most widely studied immunomodulators. Chitooligosaccharide can be prepared from the chitin or chitosan polymers through enzymatically, chemically or physically processes. Chitooligosaccharide and its derivatives have been proven to have a wide range of biological activities including intestinal flora regulation, immunostimulant, anti-tumor, anti-obesity and anti-oxidation effects. This review summarizes the latest research of the preparation methods, biological activities in immunity and safety profiles of Chitooligosaccharide and its derivatives. We recapped the effect mechanisms of Chitooligosaccharide basing on overall immunity. Comparing the effects of Chitooligosaccharide with different molecular weights and degree of aggregation, a reference range for usage has been provided. This may provide a support for the application of Chitooligosaccharide in immune supplements and food. In addition, future research directions are also discussed. COVID19, COVID19 COVID 19, 19 COVID-19 peoples people s significantly β1, β1 β 1, 1 β-(1 4linked linked 4 glucosamine, glucosamine -glucosamine furthermore immunomodulators enzymatically processes regulation immunostimulant antitumor, antitumor anti tumor, tumor anti-tumor antiobesity obesity antioxidation oxidation methods aggregation provided food addition discussed COVID1 COVID-1 β-( COVID- β-

Abstract Wetting of metal droplet on the solid substrate is a fundamental phenomenon which is applicable to the surface chemistry. When an oscillation field is included in the wetting condition, the wetting process shows significant advancing and receding behaviors. Also, the use of ultrasonic oscillation field is promising in welding. However, some odd morphologies led by the ultrasonic-treatment have shown wetting kinetics which have not been fully investigated. The high frequency ultrasonic vibration brings hysteresis to the contact angle, whose extra energy is attributed by the oscillation field. The ultrasonic wetting process is excited by the 20 kHz frequency periodic oscillation, during which droplet is swaying cyclically. However, after capturing the transformation of droplet morphologies, it is found that the frequency of each swaying cycle is identified to be 180 ms. Theoretical investigations have also quantitively proved that the energy for the contact angle decrease origins from the ultrasonic field, and the wettability is in a great enhancement. Thermal and kinetic effects of ultrasonic are investigated by making theoretical calculations, the 20 kHz ultrasonic field lasting for 5 seconds. Thermodynamics, vibrational mechanics, and interfacial phenomena affect the sonochemistry of wetting.

Abstract Background Alpha-kinase 1 (ALPK1) is a master regulator in inflammation and has been proved to promote renal fibrosis by promoting the production of IL-1β in diabetic nephropathy (DN) mice. Pyroptosis is involved in high glucose (HG)-induced tubular cells injury, characterized by activation of Gasdermin D (GSDMD) and the release of IL-1β and IL-18, resulting in inflammatory injury in DN. It is reasonable to assume that ALPK1 is involved in pyroptosis-related tubular injury in DN. However, the mechanism remains poorly defined. Methods Immunohistochemistry (IHC) staining was performed to detect the expression of pyroptosis- and fibrosis-related proteins in renal sections of DN patients and DN mice. DN models were induced through injection of streptozotocin combined with a high-fat diet. Protein levels of ALPK1, NF-κB, Caspase-1, GSDMD, IL-1β, IL-18 and α-SMA were detected by Western blot. HK-2 cells treated with high-glucose (HG) served as an in vitro model. ALPK1 small interfering RNA (siRNA) was transfected into HK-2 cells to down-regulate ALPK1. The pyroptosis rates were determined by flow cytometry. The concentrations of IL-1β and IL-18 were evaluated by ELISA kits. Immunofluorescence staining was used to observe translocation of NF-κB and GSDMD. Results The heat map of differentially expressed genes showed that ALPK1, Caspase-1 and GSDMD were upregulated in the DN group. The expression levels of ALPK1, Caspase-1, GSDMD and CD68 were increased in renal biopsy tissues of DN patients by IHC. ALPK1expression and CD68+ macrophages were positively correlated with tubular injury in DN patients. Western blot analysis showed increased expressions of ALPK1, phospho-NF-κB P65, GSDMD-NT, and IL-1β in renal tissues of DN mice and HK-2 cells, accompanied with increased renal fibrosis-related proteins (FN, α-SMA) and macrophages infiltration in interstitial areas. Inhibition of ALPK1 attenuated HG-induced upregulation expressions of NF-κB, pyroptosis-related proteins Caspase-1, GSDMD-NT, IL-1β, IL-18, α-SMA, and pyroptosis level in HK-2 cells. Also, the intensity and nuclear translocation of NF-κB and membranous translocation of GSDMD were ameliorated in HG-treated HK-2 cells after treatment with ALPK1 siRNA. Conclusions Our data suggest that ALPK1/NF-κB pathway initiated canonical caspase-1-GSDMD pyroptosis pathway, resulting in tubular injury and interstitial inflammation of DN.

ABSTRACT Insulin antibodies (IAs) induced by exogenous insulin rarely cause hypoglycemia. However, insulin autoantibodies (IAAs) in insulin autoimmune syndrome (IAS) can cause hypoglycemia. The typical manifestations of IAS are fasting or postprandial hypoglycemia, elevated insulin level, decreased C-peptide levels, and positive IAA. We report a 45-year-old male with type 1 diabetes mellitus (T1DM) treated with insulin analogues suffering from recurrent hypoglycemic coma and diabetic ketoacidosis (DKA). His symptoms were caused by exogenous insulin and were similar to IAS. A possible reason was that exogenous insulin induced IA. IA titers were 61.95% (normal: < 5%), and the concentrations of insulin and C-peptide were > 300 mU/L and < 0.02 nmol/L when hypoglycemia occurred. Based on his clinical symptoms and other examinations, he was diagnosed with hyperinsulinemic hypoglycemia caused by IA. His symptoms improved after changing insulin regimens from insulin lispro plus insulin detemir to recombinant human insulin (Gensulin R) and starting prednisone.

Los anticuerpos contra la insulina (AI) inducidos por la insulina exógena raramente causan hipoglucemia. No obstante, los autoanticuerpos contra la insulina (AIA) en el síndrome autoinmune de insulina (SAI) pueden causar hipoglucemia. Las manifestaciones típicas del SAI son la hipoglucemia en ayunas o posprandial, niveles elevados de insulina, la disminución del nivel de péptido C y AIA positivos. Presentamos un paciente hombre de 45 años con diabetes mellitus de tipo 1 (DMT1) tratado con análogos de insulina, que sufría comas hipoglucémicos recurrentes y cetoacidosis diabética (CAD). Sus síntomas fueron causados por la insulina exógena y fueron similares al SAI. La posible razón fue que la insulina exógena indujo AI. El título de AI era del 61,95% (Normal: < 5%), y las concentraciones de insulina y péptido C eran > 300 mU/L y < 0,02 nmol/L cuando se producía la hipoglucemia. Basados en sus síntomas clínicos y otros exámenes, se le diagnosticó hipoglucemia hiperinsulinémica causada por la AI. Sus síntomas mejoraron después de cambiar el régimen de insulina de lispro más insulina detemir a insulina humana recombinante (Gensulin R) y de empezar a tomar prednisona.

Abstract This study aimed to investigate the therapeutic effect of koumine on collagen-induced arthritis (CIA) in mice. Koumine was extracted from Gelsemium Elegans Benth. The CIA model was established in Balb/c mice. Forty successfully modeled mice were randomly divided into model group and 2, 4 and 8 mg/kg koumine groups, 10 mice in each group. Another 10 normal mice were selected as control group. The 2, 4 and 8 mg/kg koumine groups were treated with 2, 4 and 8 mg/kg koumine, respectively, for three successive weeks. At the end of treatment, compared with model group, in 4 and 8 mg/kg koumine groups the arthritis index was significantly decreased (P < 0.05), the peripheral blood interleukin-17 level and T helper 17 (Th17) cell percentage were significantly decreased (P < 0.05), the peripheral blood interleukin-10 level and regulatory T (Treg) cell percentage were significantly increased (P < 0.05), the spleen tissue RORγt protein expression level was significantly decreased (P < 0.05), and the spleen tissue Foxp3 protein expression level was significantly increased (P < 0.05). In conclusion, koumine has therapeutic effect on CIA in mice. The mechanism may be related to its regulating the RORγt/Foxp3 expressions, thus correcting the Th17/Treg immune imbalance.

Resumo Fundamento As células progenitoras endoteliais (CPEs) desempenham um papel importante na manutenção da função endotelial. A síndrome metabólica (SM) está associada à disfunção das CPEs. Embora o exercício físico tenha um impacto benéfico na atividade das CPEs, seu mecanismo ainda não está completamente esclarecido. Objetivo O objetivo deste estudo é investigar os efeitos do exercício físico nas funções das CPEs e os mecanismos subjacentes em pacientes com SM. Métodos Os voluntários com SM foram divididos em grupo exercício (n=15) e grupo controle (n=15). Antes e após 8 semanas de treinamento físico, as CPEs foram isoladas do sangue periférico. Foram feitos o ensaio de unidades formadoras de colônias (UFC), o ensaio de formação de tubos, a expressão proteica do óxido nítrico sintase endotelial (eNOS), da fosfatidilinositol-3-quinase (PI3-K) e da proteína quinase B (AKT). Considerou-se um valor de probabilidade <0,05 para indicar significância estatística. Resultados Após 8 semanas, o número de UFCs aumentou significativamente no grupo exercício em comparação com o grupo controle (p<0,05). Além disso, observamos uma diminuição significativa do modelo de avaliação da homeostase da resistência à insulina (HOMA-IR), endotelina-1, proteína C reativa de alta sensibilidade e dos níveis de homocisteína no grupo exercício. A intervenção com exercícios também pode aumentar a capacidade de formação de tubos de CPEs e aumentar o nível de fosforilação de eNOS, PI3-K e AKT. Conclusão O exercício físico aprimorou as funções das CPEs. O mecanismo pode estar relacionado ao exercício, ativando a via PI3-K/AKT/eNOS.

Abstract Background Endothelial progenitor cells (EPCs) play an important role in maintaining endothelial function. Metabolic syndrome (MetS) is associated with EPC dysfunction. Although physical exercise has a beneficial impact on EPC activity, its mechanism is not completely clear yet. Objective The purpose of this study is to investigate the effects of physical exercise on the functions of EPCs and the underlying mechanisms in patients with MetS. Methods Volunteers with MetS were divided into exercise group (n=15) and control group (n=15). Before and after 8 weeks exercise training, EPCs were isolated from peripheral blood. Colony forming unit (CFU) assay, tube-formation assay, the protein expression of endothelial nitric oxide synthase (eNOS), phosphatidylinositol-3-kinase (PI3-K) and protein kinase B (AKT) were determined. A probability value <0.05 was considered to indicate statistical significance. Results After 8 weeks, the number of CFUs was significantly increased in the exercise group compared to the control group (p<0.05). In addition, we observed a significant decrease of homeostasis model assessment for insulin resistance (HOMA-IR), endothelin-1, high-sensitive C-reactive protein, and homocysteine levels in the exercise group. Exercise intervention could also enhance tube-formation capacity of EPCs and increase phosphorylation level of eNOS, PI3-K and AKT. Conclusion Physical exercise enhanced the functions of EPCs. The mechanism may be related to exercise, activating the PI3-K/AKT/eNOS pathway.

The etiology of polycystic ovary syndrome (PCOS) is complex and the pathogenesis is not fully understood. Some studies have shown that dysregulation of ovarian granulosa cells may be related to abnormal follicles and excessive androgen in women with PCOS. Our team has also confirmed the high expression status of H19 in PCOS patients in the early stage. However, the relationship between H19 and miR-19b in the development of PCOS is still unknown. Therefore, we used bioinformatics to predict the binding sites of human H19 and miR-19b, and of miR-19b and CTGF genes. After the silencing and overexpression of H19, real-time polymerase chain reaction (PCR) was used to detect the expressions of H19, miR-19b, and CTGF. Western blotting was used to detect CTGF protein. Proliferation of KGN cells after H19 silencing was detected by CCK8. Flow cytometry was used to detect the apoptosis of KGN cells after H19 silencing. After the overexpression of H19, it was found that the expression of miR-19b gene decreased and the expression of CTGF increased, whereas silencing of H19 did the opposite. In addition, H19 could promote cell proliferation and decrease cell apoptosis. Finally, luciferase reporter assays showed that the 3′-end sequences of lncRNA H19 and CTGF contained the binding site of miR-19b. In conclusion, our study indicated that lncRNA H19 acted as a ceRNA to bind to miR-19b via a “sponge” to regulate the effect of CTGF on KGN cells, which may play a vital role in PCOS.

The aim of this study was to elucidate the concise effects of a traditional herb pair, Curcumae rhizoma-Sparganii rhizoma (CRSR), on uterine leiomyoma (UL) by analyzing transcriptional profiling. The UL rat model was made by intramuscular injection of progesterone and gavage administration of diethylstilbestrol. From 11 weeks of the establishment of the model, rats of the UL+CRSR group were gavaged daily with CRSR (6.67 g/kg). The serum concentrations of progesterone (P) and estradiol (E2) were determined by radioimmunoassay, the uterine index was measured by caliper measurement, and the pathological status was observed by hematoxylin and eosin stain. Gene expression profiling was checked by NimbleGen Rat Gene Expression Microarrays. The results indicated that the uterine mass of UL+CRSR rats was significantly shrunk and serum P and E2 levels significantly reduced compared to UL animals and nearly to the level of normal rats. Results of microarrays displayed the extensive inhibition of CRSR upon the expression of proliferation and deposition of extracellular matrix (ECM)-related genes, and significantly regulated a wide range of metabolism disorders. Furthermore, CRSR extensively regulated key pathways of the UL process, such as MAPK, PPAR, Notch, and TGF-β/Smad. Regulation of the crucial pathways for the UL process and ECM metabolism may be the underlying mechanisms of CRSR treatment. Further studies will provide clear clues for effectively treating UL with CRSR.

SUMMARY: Matrigel is a basement membrane matrix extracted from the EHS mouse tumor containing extracellular matrix protein, its main components are laminin, type IV collagen, nestin, heparin sulfate, growth factor and matrix metalloproteinase.At room temperature, Matrigel polymerized to form a three dimensional matrix with biological activity. It can simulate the structure, composition, physical properties and functions of the cell basement membrane in vivo, which is beneficial to the culture and differentiation of the cells in vitro, and can be used for the study of cell morphology, biochemical function, migration, infection and gene expression. In this study, Matrigel three-dimensional culture model of bone marrow mesenchymal stem cells(BMSCs) was established, and its morphology, proliferation and survival were observed. BMSCs were isolated and cultured with whole bone marrow adherence method. The Second generation BMSCs with good growth condition were selected and mixed with Matrigel to form cell gel complexes. The morphology and proliferation of mesenchymal stem cells were observed by phase contrast microscope and HE staining,Live/Dead staining was used to evaluate the cell activity.Phase contrast microscopy showed that BMSCs were reticulated in Matrigel and proliferated well, After 7 days, the matrix gel gradually became soft and collapsed, a few cell reticular crosslinking growth was seen at 14 days; HE staining showed that the cytoplasm of the cells was larger on the fourth day and the cells were elongated and cross-linked on the seventh day; Live/dead staining showed that most cells showed green fluorescence with the prolongation of culture time, on the first, 4 and 7 days, the activity of bone marrow mesenchymal stem cells in Matrigel gradually increased, and the percentages were 92.57 %, 95.54 % and 97.37 %, respectively. Matrigel three-dimensional culture system can maintain the morphology, function and proliferation ability of bone marrow mesenchymal stem cells.

RESUMEN: Matrigel es una matriz de membrana basal extraída del tumor de ratón EHS que contiene proteína de matriz extracelular. Los componentes principales son laminina, el colágeno tipo IV, nestina, sulfato de heparina, factor de crecimiento y metaloproteinasa de matriz. A temperatura ambiente, Matrigel se polimerizó para formar una matriz tridimensional. Es posible simular la estructura, la composición, las propiedades físicas y las funciones de la membrana basal celular in vivo, lo que es beneficioso para el cultivo y la diferenciación de las células in vitro, y se puede utilizar para el estudio de la morfología celular, la función bioquímica, la migración, infección y expresión génica. En este estudio, se estableció el modelo de cultivo tridimensional Matrigel de células madre mesenquimales de médula ósea (BMSC), y se observó su morfología, proliferación y supervivencia. Las BMSC fueron aisladas y cultivadas con el método de adherencia de la médula ósea completa. Se seleccionaron las BMSC de segunda generación con buenas condiciones de crecimiento y se mezclaron con Matrigel para formar complejos de gel de células. La morfología y la proliferación de las células madre mesenquimales se observaron con microscopio de contraste de fase y se tiñó con Hematoxilina-Eosina (HE); para evaluar la actividad celular se usó la tinción Live/Dead. La microscopía de contraste mostró que las BMSC se reticularon en Matrigel y proliferaron bien. Después de 7 días, se observó que el gel de matriz gradualmente se volvió blando y colapsó, y se visualizó un cruce transversal de algunas células reticulares a los 14 días. La tinción mostró que la mayoría de las células mostraron una fluorescencia verde con la prolongación del tiempo de cultivo; en los primeros 4 y 7 días, la actividad de las células madre mesenquimales de la médula ósea en Matrigel aumentó gradualmente y los porcentajes fueron de 92,57 %, 95,54 % y 97,37 %, respectivamente. El sistema de cultivo tridimensional de Matrigel puede mantener la morfología, la función y la capacidad de proliferación de las células madre mesenquimales de la médula ósea.