This study investigated the effect of puerarin on human umbilical vein endothelial cells (HUVEC) injured with hydrogen peroxide (H₂O₂). HUVEC were divided into three groups: a control group, a model group (H₂O₂ 400 μmol·L^-1) and a puerarin-treated group (3, 10, 30 and 100 μmol·L^-1). HUVEC were cultured with varied concentration of puerarin for 2 h and treated with H₂O₂ for another 24 h. Cell proliferation was detected by a CCK-8 assay. The mitochondrial membrane potential was measured by a JC-1 fluorescent probe. A transwell chamber assay was adopted to observe cell migration ability. Mitochondrial respiratory function was measured in a two-chamber titration injection respirometer (Oxygraph-2k). The expression of interleukin-1β (IL-1β), interleukin-18 (IL-18) and tumor necrosis factor-α (TNF-α) was detected by quantitative real-time PCR. The expression of pyroptosis-mediated proteins, including cleaved-cysteinyl aspartate-specific proteinase-1 (caspase-1), N-gasdermin D (N-GSDMD), NOD-like receptor protein 3 (NLRP3) and purinergic ligand-gated ion channel 7 receptor (P2X7R) was detected by Western blot. The results show that 400 μmol·L^-1 H₂O₂ treatment for 24 h causes obvious damage to HUVEC. Compared with the model group, puerarin protected against cellular injury in a dose-dependent manner, with the greatest effect at a dose of 30 and 100 μmol·L^-1. Puerarin significantly decreased the mitochondrial membrane potential and improved mitochondrial function. Puerarin inhibited cell migration induced by H₂O₂, suppressed the expression of IL-1β, IL-18 and TNF-α, and down-regulated the pyroptosis-mediated protein. These changes are statistically significant (P < 0.05). These findings demonstrate that puerarin has a protective effect against H₂O₂-induced oxidative damage of HUVEC by inhibiting the migration of HUVEC cells. The mechanism may be related to improved mitochondrial respiratory function and inhibition of pyroptosis.

This study was to investigate the protective effects of puerarin on myocardial ischemia/reperfusion (MI/R) injury and the underlying mechanism. The MI/R-model was established by ligating the left anterior descending artery (LAD) for 60 min followed by 24 h reperfusion, puerarin (10, 30, and 100 mg·kg^-1) was orally administered 20 min before reperfusion. Cardiac function, myocardial infarct index, cardiac damage markers, inflammatory cytokines, and apoptosis index were measured to evaluate the protective effects of puerarin on MI/R injury. The activation of Nod-like receptor protein 3 (NLRP3) inflammasome and Toll like receptor 4 (TLR4)/myeloid differentiation factor 88 (Myd88)/nuclear factor kappa B (NF-κB) pathway were determined by Western blot. All animal experimental procedures were approved by the ethics committee of the Institute of Materia Medica, Peking Union Medical College, Chinese Academy of Medical Sciences. The results showed that puerarin could significantly improve cardiac function, reduce myocardial infarct size, decease the levels of lactic dehydrogenase (LDH), aspartate transaminase (AST), creatine kinase-MB (CK-MB), and cardiac troponin T (cTnT) and suppress cardiomyocyte apoptosis. Meanwhile, puerarin could notably decrease the levels of inflammatory cytokines such as interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α). Western blot analysis revealed that puerarin could downregulate the expression of TLR4, Myd88, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cleaved-caspase 1, cleaved-gasdermin-D (GSDMD), IL-1β, and IL-18, as well as the phosphorylation levels of inhibitor of NF-κB α (IκBα), IκB kinase β (IKKβ), and NF-κB. These findings demonstrated that puerarin could alleviate MI/R injury by suppressing NLRP3 inflammasome activation, possibly via TLR4/Myd88/NF-κB pathway.

OBJECTIVETo compare the sensitivity and practicability of modified Bethesda assay and Nijmegen assay in detecting factor VIII (FVIII) inhibitor.

METHODSModified Bethesda assay and Nijmegen assay were used to screen FVIII inhibitors in 237 patients with hemophilia A. The buffer plus universal coagulation reference plasma (UCRP) was used to establish a standard curve for FVIII: C assay in modified Bethesda method, instead of Nijmegen plasma plus FVIII deficiency plasma in Nijmegen method. The cutoff value for positive FVIII inhibitors is > or = 0.6 BU/ml.

RESULTSThe positive rate of FVIII inhibitors was 5.5% (n = 13) when using modified Bethesda assay and was 8.4% (n = 20) when using Nijmegen assay (P > 0.05).

CONCLUSIONModified standard Bethesda assay is a convenient and feasible method for detecting FVIII inhibitors.

Adolescent ; Adult ; Autoantibodies ; blood ; Blood Coagulation Tests ; methods ; Child ; Child, Preschool ; Factor VIII ; immunology ; Female ; Hemophilia A ; blood ; immunology ; Humans ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult

OBJECTIVETo explore cell death and apoptosis in rat hippocampal neurons at different time points after ischemia, hypoxia and reperfusion injury and to elucidate time window characteristics in ischemia neuronal injury.

METHODSHippocampal neurons were obtained from rat embryo and were cultured in vitro. The ischemia and reperfusion of cultured rat hippocampal neurons were simulated by oxygen-glucose deprivation (OGD) and recovery. OGD at different time points (0.25 h to 3.0 h) and then the same recovery (24 h) were prepared. Annexin V-PI staining and flow cytometry examined neuron death and apoptosis at different time after injury.

RESULTSAfter OGD and recovery, both necrosis and apoptosis were observed. At different times after OGD, there were statistically significant differences in neuron necrosis rate (P < 0.05), but not in apoptosis rate (P > 0.05). At recovery, survival rate of hippocampal neurons further decreased while apoptosis rate increased. Furthermore, apoptosis rates of different time differed greatly (P < 0.05). Apoptosis rate gradually increased with significant difference among those of different time points (P < 0.05). However, 2 h after ischemia, apoptosis rate decreased markedly.

CONCLUSIONSApoptosis is an important pathway of delayed neuron death. The therapeutic time window should be within 2 h after cerebral ischemia and hypoxia.

Animals ; Apoptosis ; physiology ; Brain Ischemia ; pathology ; Cell Death ; physiology ; Cell Hypoxia ; Cells, Cultured ; Disease Models, Animal ; Female ; Fetus ; cytology ; Flow Cytometry ; Hippocampus ; pathology ; Neurons ; pathology ; Pregnancy ; Pregnancy, Animal ; Probability ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; pathology ; Sensitivity and Specificity ; Time Factors

Objective To observe Th-1 drift induced in vitro by high immunogenic glioblastoma multiforme (GBM) U251 cell vaccine with high expression of membrane-enriched heat shock protein 70 (Hsp70) and major histocompatibility complex class Ⅰ (MHC-Ⅰ) molecules. Methods The high expression of MHC-Ⅰ and Hsp70 in U251 cells were induced by 500 U/ml IFN-γ for 48 h, heat shock at 43 ℃ for 2 h, or their combination. The cells were then inactivated by the mitomycin C (MMC) to prepare the cell vaccine. Peripheral blood mononuclear cells (PBMCs) from healthy donators were incubated with GBM U251 cell vaccines as the effector cells. Flow cytometry was applied to analyze the changes of CD4<'+> and CD8<'+>T lymphocytes in the PBMCs. The secretion of IFN-γ and IL-2 of the effector cells, after assaulting the target cells, was evaluated by ELISA. Results The percentages of CD4<'+> and CD8<'+> T lymphocytes of the PBMCs incubated with the U251 cell vaccine increased significantly as compared to that stimulated by the membrane-enriched MHC class Ⅰ or Hsp 70 molecule U251 cell vaccines (P<0.05), and so was the secretion of IFN-γ and IL-2 (P<0.05). Conclusions Th-1 drift stimulated by GBM U251 cell vaccine with high immunogenicity, high expression of Hsp 70 and membrane-enriched MHC class Ⅰ molecules plays an important role in antitumor mechanism in vitro.

Objective To investigate the rehabilitation promoting factors in stroke patients and to provide references for the design and implementation of effective intervention for rehabilitation of stroke patients.Methods In-depth interviews were conducted among eight stroke patients,and interview data were collected and analyzed.Results Five themes were identified through analysis and classification of the interview data:practical rehabilitation goals,effective rehabilitation training behaviors,overcoming abandonment behaviors and negative emotions,suitable support system,and proper self-adjustment.Conclusion The rehabilitation promoting factors for stroke patients are performing effective rehabilitation training towards effective rehabilitation goals.In this process,patients need to rely on appropriate social support and patients' self-adjustment to overcome abandonment behaviors and negative emotions.These factors form a force to promote rehabilitation during the process of rehabilitation.

Objective To investigate and compare the curative effect between delayed percutaneous coronary intervention (PCI) for patients with acute myocardial infarction presenting 12-24 hours from symptom onset and medical therapy on acute myocardial infarction patients presenting with ST-segment elevation (STEMI). Methods Using a prospective,open,parallel,controlled research approach,186 patients with STEMI were divided into delayed PCI group(n=89),which received PCI within 12-24 hours after STEMI and medical therapy group(n=97),which received medical therapy after STEMI. All patients were followed up 1-6 months with average follow-up (5.6 ± 1.4) months. Data of hospitalization period, the cardiac structures detected by echocardiography such as left atrial diameter (LAD), left ventricular diastolic diameter(LVDd),left ventricular ejection fraction LVEF,left ventricular fractional shortening(LVFS),composite end point events and major adverse cardiac events(MACE)were compared between the two groups.Results Compared with medical therapy group, the hospitalization cycle was significantly shorter in delayed PCI group. Data of the LAD and LVDd were significantly decreased,but LVEF and LVFS were increased in delayed PCI group compared with those of medical therapy group at 30 d and 6-month follow-up. The incidence of MACE and composite end point events were significantly less in delayed PCI group than those of medical therapy group (P<0.05). Conclusion Delayed PCI treatment can decrease the time of hospital stay and decrease the incidence rates of MACE and composite end point events,and improve left ventricular function and prognosis of patients.

OBJECTIVECarnitine deficiency has been associated with progressive cardiomyopathy due to compromised energy metabolism. The objective of this study was to investigate clinical features of carnitine deficiency-induced cardiomyopathy and the therapeutic efficacy of L-carnitine administration.

METHODBetween January 2010 and December 2011, filter-paper blood spots were collected from 75 children with cardiomyopathy. Free carnitine and acylcarnitine profiles were measured for each individual by tandem mass spectrometry (MS/MS). For those in whom carnitine deficiency was demonstrated, treatment was begun with L-carnitine at a dose of 150 - 250 mg/(kg·d). Clinical evaluation, including physical examination, electrocardiography, chest x-ray, echocardiography and tandem mass spectrometry, was performed before therapy and during follow-up.

RESULTOf 75 cardiomyopathy patients, the diagnosis of carnitine deficiency was confirmed in 6 patients, which included 1 boy and 5 girls. Their age ranged from 0.75 to 6 years. Free carnitine content was (1.55 ± 0.61) µmol/L (reference range 10 - 60 µmol/L). Left ventricular end-diastolic diameter (LVDd) was (5.04 ± 0.66) cm and left ventricular ejection fraction (LVEF) was (38.5 ± 10.5)%. After 10 - 30 d therapy of L-carnitine, free carnitine content rose to (30.59 ± 15.02) µmol/L (t = 4.79, P < 0.01). LVDd decreased to (4.42 ± 0.67) cm (t = 4.28, P < 0.01) and LVEF increased to (49.1 ± 7.6)% (t = 6.59, P < 0.01). All patients received follow-up evaluations beyond 6 months of treatment. Clinical improvement was dramatic. LVEF returned to normal completely in all the 6 patients. LVDd decreased further in all the 6 patients and returned to normal levels in 3 patients. No clinical signs or symptoms were present in any of the 6 patients. The only complications of therapy had been intermittent diarrhea in 1 patient.

CONCLUSIONTandem mass spectrometry is helpful to diagnose carnitine deficiency and should be performed in all children with cardiomyopathy. L-carnitine has a good therapeutic effect on carnitine deficiency-induced cardiomyopathy.

Adolescent ; Cardiomyopathies ; diagnosis ; drug therapy ; etiology ; Cardiotonic Agents ; administration & dosage ; therapeutic use ; Carnitine ; blood ; deficiency ; therapeutic use ; Child ; Child, Preschool ; Electrocardiography ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Retrospective Studies ; Tandem Mass Spectrometry ; Treatment Outcome ; Ventricular Function, Left ; drug effects

OBJECTIVETo study the newborn's Apgar score in 'one minute' and relevant factors.

METHODSOne year inpatient woman from a Maternal and Child Health Hospital of Anhui province were selected by cluster sampling method and newborn asphyxia situation was investigated using Apgar score and self-designed questionnaire.

RESULTSThe Apgar score in 'one minute' which marking 8 to 10, 4 to 7 and 0 to 3 were found in 1875 (73.78%), 426 (16.77%) and 240 infants (9.45%) respectively. The average Apgar score in 'one minute' and five minutes were (7.69 +/- 2.27) and (9.01 +/- 1.89) respectively, The Apgar score in 'one minute' was significantly correlated with that of five minutes (Pearson coefficient correlation r = 0.841, P = 0.00). Ordinal regression analysis revealed that parturient age (OR = 1.04), being farmer (OR = 2.22), parity (OR = 1.26), assistant vaginal delivery (OR = 4.93), caesarean section (OR = 1.95), pregnancy-induced hypertension syndrome (OR = 1.42), albuminuria in gestational period (OR = 1.44), newborn being male (OR = 1.23), low birth weight (OR = 2.94), inborn abnormality (OR = 12.12), premature birth (OR = 1.22) and complications of delivery (OR = 5.04) were risk factors while the number of years under study (OR = 0.91), prenatal check-up (OR = 0.48), body length of newborn infant (OR = 0.88) and single birth (OR = 0.57) were protective factors.

CONCLUSIONApgar score in 'one minute' of newborn infant was affected by several factors as stated above. Health care program in earlier period toward community parturient should be strengthened in order to discover and control high risk factors of duration of pregnancy in earlier period.

Apgar Score ; Asphyxia Neonatorum ; epidemiology ; Epidemiologic Studies ; Female ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Risk Factors ; Time Factors

OBJECTIVETo discuss the experiences of diagnosis and treatment for vasoactive intestinal peptide-secreting-tumors (VIPoma) by summarizing clinical informations of 15 patients with VIPoma.

METHODSTo analyze Clinical manifestations, laboratory examinations, imaging features, operation, pathological findings and follow up survey of 15 patients, among them 1 case from our hospital and the other 14 cases were searched in chinese biological and medical literature database from Jan 1987 to Dec 2002.

RESULTSThe main clinical manifestation include periodical secretory watery diarrhea, hypokalemia, achlorhydria, in addition, periodical backache, skin rash, and polyps of colon were presented in the case in our hospital. The immunohistochemical expression of many kinds of digestive hormone including VIP presented positive; All clinical symptoms of which except polyps disappeared after operation, elevated VIP data in serum also markedly decreased. Part resection of superior mesenteric vein was performed in the same patient.

CONCLUSIONSVIPoma is rare. Typical symptoms and the serum value of VIP were keys to diagnosis, the operation is the most effective means for treatment. Resection of tumor, Radiofrequency tissue ablation, liver transplantation can be selected for metastatic VIPoma in the liver.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; diagnosis ; pathology ; therapy ; Vasoactive Intestinal Peptide ; secretion ; Vipoma ; diagnosis ; pathology ; therapy