Child ; Female ; Humans ; Hydroa Vacciniforme ; Hypersensitivity ; Insect Bites and Stings

Animals ; Centrifugation, Density Gradient ; methods ; Chickens ; Cytochrome P-450 Enzyme System ; analysis ; Microsomes, Liver ; chemistry ; ultrastructure ; Sucrose

Objective To investigate the changes of expression of Fas-associated proteins named Fas-associated death domain protein(FADD)and death-associated protein(Daxx)in the ischemic penumbra following transient focal cerebra ischemia in rats.Methods ①Adult male Sprague-Dawley rats were randomly divided into the sham-operated group and the cerebral ischemia model group.Rats underwent right middle cerebral artery occlusion(MCAO)for 2 h and reperfusion for 1,3,6,12 and 24 h using an intraluminal suture technique.The expression of FADD and Daxx mRNA and protein were measured with methods of immunohistochemistry.Western blot and reverse transcription-polymerase chain reaction(RT- PCR)respectively were used in the ischemic penumbra of rats.②Double-label fluorescence confocal laser scanning microscopy(CLSM)was performed to monitor FADD and Daxx intracellular location before and after ischemia.Results RT-PCR,Immunohistochemistry,Western blot experiments indicated that a very low level of FADD mRNA and protein were detected in the cerebral cortex of sham rats.The expression level both of FADD mRNA and protein increased significantly at 3 h after reperfusion,peaked at 12 h,then declined markedly at 24 h in the ischemic penumbra of model rats.RT-PCR,Immunohistochemistry indicated that a relatively high level of Daxx mRNA was detected in the cerebral cotex of sham rats.The expression level of Daxx mRNA increased significantly at 3 h after reperfusion and persisted to 24 h at a high level,whose protein had a same change of expression level in the ischemic penumbra of model rats. Immunofluorescence double-staining laser scanning by CLSM showed that the immunoreactivity of FADD was located in cytoplasm,and the intracellular translocation of the immunoreactivity of Daxx from nucleus to cytoplasm was monitored by measuring the green fluorescence after ischemia.Conclusion The transient upregulation of FADD and the persistant high level of expression of Daxx may contribute to neuronal apoptosis following cerebral ischemia/reperfusion.

Autophagy is an active research area in the biomedical field as its role has been identified in many physiological and pathological processes. Accordingly, there is a growing demand to identify, quantify and manipulate the process accurately. Meanwhile, there is great interest in identifying compounds that modulate autophagy because they may have applications in the treatment of a variety of autophagy-related diseases. In this review, we summarize the current status of autophagy screening systems to facilitate identification of autophagy modulators.
Autophagy ; Humans

Biomarkers, Tumor ; blood ; Breast Neoplasms ; blood ; diagnosis ; Female ; Humans ; Protein Array Analysis ; methods ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

Objective To investigate the variation law of apparent diffusion coefficient (ADC) and fractional anisotropy (FA) values in patients with cerebral infarction, and to explore the relationship between the changes and the prognosis of cerebral infarct patients. Methods Sixteen patients with cerebral infarction were recruited and divided into 2 groups:good recovery and poor rehabilitation. ADC and FA values were calculated in infarct areas and control areas which were the regions with symmetrical position and the same area as infarct areas. The difference of ADC and FA values in patients at acute and earlier chronic stage between the two areas were analyzed. Results ①At acute stage, ADC values in infarct areas were lower than those in control areas (P<0.05). At early chronic stage, there was no significant difference of ADC values between infarct areas and control areas (P>0.05), moreover ADC values were higher than that at acute phase (P<0.05). ②FA values in infarct areas were lower than those in control areas at both acute and early chronic stage (P<0.05). At early chronic stage, FA values were lower than those at acute stage (P<0.05). ③There was no significant difference of ADC and FA values at both acute and early chronic stage between good recovery group compared with poor rehabilitation group (P>0.05). Conclusion There are certainly rules in changes of ADC and FA values in patients with cerebral infarction at acute and earlier chronic stage.

Objective To evaluate the influence of gestational diabetes mellitus (GDM) on maternal and perinatal outcomes in twin pregnancies. Methods We retrospectively analyzed the clinical features of both twin and singleton pregnancies, which delivered in Guangzhou Women and Children's Medical Center between January 1, 2012 and December 31, 2013. The twin pregnancies were divided into two groups:those with (GDM-T, n=51) and without GDM (non-GDM-T, n=130), which were matched by maternal age and delivery time (within one month) in a ratio of 1∶2 among singleton pregnancies with (GDM-S, n=102) and without GDM (non-GDM-S, n=102), respectively. The differences of adverse maternal and perinatal outcomes among these four groups were examined. The overall assessment of pregnancy outcomes was completed using Delphi method. Statistical analysis was performed with one-way analysis of variance, t test, Kruskal-Wallis test, rank test, Chi-square test or Fisher's exact test. Results (1) When compared to GDM-S and non-GDM-S group respectively, less women conceived with the help of assisted reproductive technology, higher proportion of women underwent and gestational age at delivery tend to be earlier in GDM-T and non-GDM-T group (all P<0.01). In oral glucose tolerance test,the fasting blood glucose level of GDM-T group was higher than the other three groups (F=21.716, P<0.01), the glucose levels at 1 and 2 h were higher than non-GDM-T and non-GDM-s respectively (both P<0.01), but no significant difference was found when compared with GDM-S group (P>0.01). Similarly, no significant difference was found in prenatal glycosylated hemoglobin value between GDM-T and GDM-S group (P>0.01). (2) There was no significant difference in the incidences of hypertensive disorders of pregnancy, anemia, premature rupture of membranes, oligohydramnios, placental abruption, postpartum hemorrhage, asphyxia neonatorum, small for gestational age, hypoglycemia of newborn, hyperbilirubinemia of newborn and perinatal death between GDM-T group and the other three groups(all P>0.01). Higher incidences of hypertensive disorders of pregnancy and postpartum hemorrhage were shown in the GDM-T group than in the GDM-S and non-GDM-S groups, respectively (both P<0.01). The incidences of preterm birth in GDM-T and non-GDM-T group were both higher than that in GDM-S and non-GDM-S, respectively [54.9%(66/102), 53.8%(140/260), 5.0%(10/102) and 3.0%(6/102), all P<0.01], while no significant difference was found between GDM-T and non-GDM-T group (P>0.01). (3) The overall assessment of pregnancy outcomes did not show any difference between GDM-T group and the other three groups (χ2=6.707, P>0.01). However, the score for fetal outcomes in the GDM-T group was higher than in the GDM-S and non-GDM-S group, but lower than in non-GDM-T group [M(Q)=1.0(2.3), 0.0(3.0), 0.0(0.0), 1.0(2.8) score, χ2=122.818, P<0.01]. Conclusions GDM does not increase the risk of adverse pregnant outcomes in twin pregnancies.

Objective To construct the lentivirus carrying human ?-catenin-EGFP(enhanced green fluorescent protein)and observe its expression in human follicle stem cells.Methods The ?-catenin gene sequence was amplified by RT-PCR from extraction of total RNA of human vascular endothelial cells.TA cloning technique was utilized to acquire gene subcloned pUCm-T-?-catenin.After transformation reaction,candidate clone was further analyzed by PCR and gene sequencing.Then the plasmid was transfected into FT293 cells.After identification by Western blotting,the plasmid was transfected into FT293 cells again for packaging.Infection titer was monitored by green EGFP expression.The expression of ?-catenin-lentivirus in human follicle stem cells were observed under inverted fluorescence microscope.Results The ?-catenin gene was cloned into the lentivirus successfully.The high expression of green fluorescence protein in FT293 cell line was found under fluorescent microscope.Viral titer checked by real-time PCR was about 2.0?108 TU/mL.When the multiplicity of infection(MOI)was 10,the infection efficiency of ?-catenin-lentivirus in human follicle stem cells was nearly 80% after infection 48 h around.After 3 weeks of continuous observation,we found the infection efficiency still keeping in the range of 80%-90%.Conclusion The lentivirus expression vector for ?-catenin was successfully constructed.It can steadily infect human follicle stem cells and the infection efficiency is considerable high.

Objective To evaluate the myocardial blood supply in patients with metabolic syndrome (MS) using 99Tcm-MIBI SPECT MPI. Methods A total of 342 patients were divided into four groups according to the number of abnormal metabolic indices: no abnormal metabolic index (Group 1), one abnormal index (Group 2), two abnormal indices (Group 3), three or more abnormal indices (Group 4). Each patient underwent two-day protocol of gated stress and rest 99Tcm-MIBI MPI. One hundred and three of the 342 patients were clinically diagnosed as MS and underwent CAG within 1 month after MPI. χ2test was used to evaluate the difference among the four groups and Kappa test to analyze the correlation between MPI and CAG. Results Compared with CAG, the diagnostic sensitivity, specificity, positive and negative predictive values by 99Tcm-MIBI SPECT MPI for coronary artery diseases (CAD) in 103 MS patients were 80.5% (33/41), 85.5% (53/62), 78.6% (33/42) and 86.9% (53/61), respectively. The correlation coefficient between MPI and CAG was 0.657 (P<0.001). The abnormal MPI rates in group 1, 2, 3 and 4 were 23.3% (10/43), 32.9% (26/79), 54.4% (56/103), and 57.3% (67/117), respectively (χ2=23.22, P<0.001). Conclusions In MS patients,99Tcm-MIBI SPECT MPI can be useful for evaluating myocardial blood supply and the myocardial ischemia rates may correlate positively with the number of abnormal metabolic indices.

Objective To study the expression and change of neuropeptide substance P (SP) during the wound healing of deep partial thickness scalding in diabetic rats. Methods Eighty-four Wistar rats were randomly divided into diabetes mellitus group (n=42) and control group (n=42). Diabetic rat models were established by intraperitoneal injection of streptozotocin (STZ) in diabetes mellitus group, and those in control group were intraperitoneally injected with aseptic citrate buffer solution. Deep partial thickness scalding with diameter of 2 cm on the back were prepared in all the rats. The pre-scalding and post-scalding wound specimens of different time points were obtained, and the percentages of wound closure were calculated. The wound specimens were also obtained for immunohistological staining to compare the areas with positive staining of SP, and ELISA was employed to detect the expression of SP in the wound tissues. Results The percentage of wound closure was significantly lower in diabetes mellitus group than that in control group from 7 days post-scalding (P< 0.01). The areas with positive staining of SP in diabetes mellitus group were much smaller than those in control group at different time points, which was most significant on the seventh day post-scalding[(1 350.93±99.28) μm2 vs(1 715.86± 103.41) μm2](P < 0.01). The expression of SP in the wound tissues was significantly lower in diabetes mellitus group than that in control group at different time points, which was most significant on the seventh day post-scalding[(114.04±9.96) vs(143.39±8.94)](P<0.01). Conclusion The significantly lower expression of SP in wound site may be one of the causes of delayed wound healing in diabetic rats.