Objective:Cardiogenesis is a precise process controlled by sequential gene regulatory steps.Heart-specific transcription factors (TFs)GATA4,Nkx2.5,MEF2C and Tbx5 play critical roles in controlling heart development.But the genetic basis for the temporal expression of genes during heart development still remains unclear.Substantial studies displayed gene regulation partly attributes to histone acetylation.However,the functions of individual histone acetyltransferases (HATs)in specific developmental transcription programs are not well elucidated.p300,a histone acetyltransferase and coactivator,plays key role in many physiological processes,moreover our previous study suggests that expression of p300 is developmentally regulated during mouse cardiogenesis,but the underlying molecular mechanism remains elusive.Methods:The whole hearts from embryonic mice on embryonic days (E)10.5 and 16.5 were collected accordingly,and then total RNA was extracted.Heart-specific TFs GATA4,Nkx2.5,MEF2C and Tbx5 mRNA from mouse myocardium at differential embryonic days during cardiac development were analyzed by quantitative (Q)-PCR.The levels of histone H3 acetylation on these heart-specific TFs promoter and transcription factor binding sites of p300 to these genes were identified by chromatin immunoprecipitation assays (ChIP).Results:The present study furnishes a comparative temporal expression map of cardiac transcriptional factors,during murine heart development (E10.5 and E16.5).The mRNA levels of these genes present dynamic change.On E10.5,the expressions of GATA4 and MEF2C were remarkable lower than those on E16.5 (P

Objective:To express and secrete rBPI m23 bactericidal protein in Pichia pastoris expression system. Methods:pPICZ? synBPI m600 recombinant expression vector was constructed with prepared vectors (pUC18 synBPI 414 and PBV220 BPI m420 ) according to Molecular Cloning Laboratory Manual. The Pichia pastoris GS115 were electroporated with linearized pPICZ? synBPI m600 vectors , positive clones were screened on low salt LB medium with Zeocin , then the positive clones were identified by PCR and Mut phenotype analysis. rBPI m23 was induced to express in BMMY ,purified by SP Sepharose beads , identified by SDS PAGE and Western blot and measured with BCA method. Results:①pPICZ? synBPI m600 expression vector was successfully constructed.②Pichia pastoris GS115 strains integrated with synBPI m600 stably were obtained.③SDS PAGE showed the molecular weight of the recombinant protein was approximate 23 kD,and Western blot confirmed that the protein could bind to the anti BPI antibody specially.④The supernate protein concentration was about 2 9 mg/L.Conclusion:rBPI m23 was expressed and secreted effectively in Pichia pastoris GS115.

BACKGROUND: Fresh amniotic membrane has been extensively used in treating ocular surface disease, which can inhibit fibrous tissue proliferation, inhibit neovascularization, and relieve inflammation. However, its application in treating glaucoma is rarely reported.OBJECTIVE: To explore the antiproliferative effect of fresh amniotic membrane on postoperative scar and compared with preserved amniotic membrane.METHODS: A total of 36 New Zealand white rabbits were randomly divided into 3 groups. Fresh amniotic membrane and preserved amniotic membrane combined with trabeculectomy separately underwent fresh amniotic membrane and preserved amniotic membrane transplantation. The control group was subjected to trabeculectomy alone. After 1, 2, 3, and 4 weeks, the morphology and function of filtering bleb were checked, and the platelet-derived growth factor (PDGF) of surrounding tissue was determined by immunohistochemical method.RESULTS AND CONCLUSION: Fresh amniotic membrane and preserved amniotic membrane groups displayed bulged filtering bleb with good filtering function. Pathological observation showed that fibroblasts of the filtration pathway had less, but more sparse scar tissues than control group, but inflammatory infiltration was observed in all groups. The cavity of different sizes and shapes were detected in filtration pathway of control group, which was replaced by fibrous tissue hyperplasia, with a large number of fibroblasts. Immunohistochemical results showed that the expression of PDGF was significantly less in fresh amniotic membrane and preserved amniotic membrane groups compared with the control group, and the PDGF expression was less in fresh amniotic membrane group than the preserved amniotic membrane group. Fresh and preserved amniotic membrane can improve the filtering bleb function, reduce scar formation, and maintain the patency of filtering pathway. Moreover, the effects of fresh amniotic membrane is better than preserved amniotic membrane.

Objective To enhance the utilization rate of large medical equipment in medical university.Methods The running and utilization rates of 36 pieces of large medical equipment were statistically analyzed during the past 5 years in the Scientific Experiment Center of some university,and the histogram was established for the effective machine hours to explore the utilization of large medical equipment.Results The statistical analysis and histogram showed that the utilization rate in the past 5 years was low and idleness occurred in some equipment sometimes.Conclusion Some measures are proposed to enhance the utilization rate of large medical equipment from the aspects of equipment maintenance and calibration,propaganda,new function development,personnel and etc.

Objective To observe the effects of Qingre Paidu Capsule on NF-κB and IL-1βin acute gouty arthritis model rats induced by monosodium urate crystals (MSU);To explore its preventive and therapeutic mechanism for acute gouty arthritis. Methods Sixty SD rats were randomly divided into normal group, model group, diclofenac sodium group, and Qingre Paidu Capsule high, medium and low dose groups. Rats in different treatment groups were given corresponding medicines by gavage for 7 days. On the fifth day, after 1 hour of gastric perfusion, acute gouty arthritis rat model was established by MSU injection in ankle joint cavity. All rats were sacrificed and materials were taken 48 h after the model was established. The morphological changes in the synovial tissue of ankle joint were observed by light microscope. The expression levels of NF-κB and IL-1β in the articular leachate were monitored by ELISA. Results The expression levels of NF-κB and IL-1βin the articular leachate of Qingre Paidu Capsule each dose group and diclofenac sodium group were obviously lower than the model group (P<0.05, P<0.01). Histopathologic examination revealed that Qingre Paidu Capsule could abate tissue swelling of ankle joints, reduce inflammatory cell infiltration of synovial tissues and improve the pathologic changes of the hyperplasia of synovium and so on. Conclusion Qingre Paidu Capsule play the anti-inflammatory role by inhibiting the expressions of NF-κB and IL-1βin articular tissue of rats.

Objective To study the effects of mild hypothermia on cognitive function , synapsinⅠexpression and synaptic ultrastructure of hippocampus in epileptic rats induced by global cerebral ischemia. Methods Forty-eight male SD rats were randomly divided into control (NC) group, sham-operated (Sham) group, normothermic epilepsy (NT) group and mild hypothermic epilepsy (HT) group. The model of postischemic audio-genetic seizure was established by chest compression. Hypothermia intervention was given to HT group. Immunocytochemistry was conducted to detect the expressions of synapsin I in hippocampus at days 1 , 3, 14. the synaptic ultrastructure and cognitive function were respectively observed by electron microscope and Morris water maze. Results Compared with NC and Sham group, the expression of synapsinI in NT group was decreased, the escape latency was prolonged and across platform number decreased (P < 0.05). The synapses were decreased in number, and mitochondria was viewed swelling, synaptic membranes unclear, myelin fractured. Compared with NT group, the expression of synapsinⅠin HT group had no obvious change in 24 h but was significantly increased in days 3 and 14 (P < 0.01); The escape latency was decreased and the number of cross platform increased (P < 0.01); Synaptic structure was clear, with interface growing and postsynaptic density thickened. Conclusion Mild hypothermia may improve the cognitive function of the epileptic rats induced by global cerebral ischemia by upregulating the expression of synapsinⅠand alleviating the damage of synaptic structure.

ObjectiveTo analyze the clinical manifestation of hemolytic disease of the newborn (HDN) due to anti-M and Rhesus system.MethodsClinical information was collected and analyzed for three cases with HDN due to anti-M and 64 with Rhesus hemolytic disease, who were admitted to Department of Neonatology, Beijing Children's Hospital Affiliated to Capital Medical University from February 2011 to January 2015, as well as another 28 cases of HDN due to anti-M with complete information retrieved from literature in Wanfang and China National Knowledge lnfrastructure (CNKI) Database from 1992 to 2014.Chi-square test was performed for statistical analysis.ResultsTwo out of the 64 Rh hemolytic babies gave up therapy due to kernicterus and another two out of the 31 MN hemolytic babies, obtained from literature, died 24 h after birth because of anemia or edema, while the rest survived. Although more babies were the first child of the family in HDN due to anti-M than those of Rh hemolytic disease [26%(8/31) vs 9%(6/64),χ2=4.487, P=0.034], but lower incidence of jaundice [81%(25/31) vs 98%(63/64),χ2=9.686,P=0.002], less proportion of presentation of jaundice within 24 h after birth [29% (9/31) vs 64%(41/64),χ2=10.279,P=0.001] and lower positive rate of direct antiglobulin test [39%(12/31) vs 100%(64/64), Fisher exact test,P=0.000] were shown in HDN due to anti-M. No significant difference was found in the incidences of hyperbilirubinemia [58%(18/31) vs 66%(42/64),χ2=0.513], severe hyperbilirubinemia [23%(7/31) vs 36%(23/64),χ2=1.724], anemia [81%(25/31) vs 89%(57/64),χ2=1.253] and severe anemia [29%(9/31) vs 34%(22/64),χ2=0.271] between HDN due to anti-M and Rh hemolytic babies (allP>0.05).ConclusionsHDN due to anti-M and Rhesus hemolytic disease can cause severe pathological jaundice and/or anemia in newborns. Indirect antiglobulin test should be offered when direct antiglobulin test is negative which is helpful in the diagnosis of HDN due to anti-M.

Objective:To investigate the clinical characteristics and prognosis of cytomegalovirus (CMV) reactivation in patients with liver failure.Methods:A total of 75 patients diagnosed with liver failure and tested for serum CMV DNA between January 2016 and June 2019 in Huashan Hospital, Fudan University were retrospectively analyzed. According to the CMV DNA test results, the patients were divided into CMV DNA positive group and CMV DNA negative group. The classification of liver failure, the use of glucocorticoids, the proportions of T lymphocyte subsets of the two groups were compared and the prognosis was evaluated. Mann-Whitney U test and chi-square test were used to analyze the data. Results:Of the 75 patients with liver failure, 17 were CMV DNA positive and 58 were CMV DNA negative. Among the 17 CMV DNA positive patients, nine were acute (subacute) liver failure, and 13 were treated with glucocorticoids, which were all significantly higher than those in the CMV negative group (20.7%(12/58) and 20.7%(12/58), respectively). The differences were both statistically significant ( χ2=6.70 and 18.40, respectively, both P<0.05). The proportions of CD3 + T lymphocytes and CD8 + T lymphocytes in the CMV DNA positive group were both higher than those in the CMV DNA negative group, and the proportions of CD4 + T lymphocytes, the ratio of CD4 + /CD8 + T lymphocytes and the proportion of B lymphocytes were all lower than those in the CMV DNA negative group. The differences were all statistically significant ( U=274.50, 165.50, 273.00, 185.00 and 189.00, respectively, all P<0.05). Acute (subacute) liver failure (odds ratio ( OR)=4.3, 95% confidence interval ( CI) 1.3-12.6) and glucocorticoid use ( OR=12.5, 95% CI 3.4-38.3) were risk factors for CMV reactivation in patients with liver failure. The disease improvement rate in the CMV DNA negative group was 56.9% (33/58), and five out of 17 patients improved in the CMV DNA positive group, with a statistically significant difference ( χ2=1.99, P=0.04). Conclusions:The use of glucocorticoids increases the risk of CMV reactivation in patients with liver failure, and CMV reactivation in patients with liver failure presents immune disorders which seriously affect their prognosis. Therefore, it is important to pay attention to CMV DNA monitoring in patients with liver failure using glucocorticoids.

Objective To understand and compare the proportion of neural stem cells (NSCs) in the whole brain and cerebral cortex of mice at different embryonic days, and provide quantitative data for the later optimization of NSCs isolation and culture.Methods The whole brains (at embryonic 12.5, 14, 16 and 18 days) and cerebral cortex (at embryonic14, 16 and 18 days) were isolated and digested into single cell suspension, and were adherently cultured for 3-4 h.Immunofluorescence staining of Nestin, a NSCs specific marker, was used to statistically analyze the proportion of NSCs in each group.Expression of Nestin mRNA in the cerebral cortex of mice at E12.5, E14, E16, and E18 was detected by real-time fluorescence quantitative PCR.Results The result of immunofluorescence assay showed that there were Nestin-positive cells in the whole brain and cerebral cortex of mice at different embryonic days.In the whole brain,the proportion of NSCs was highest at E12.5 (53.42±1.57%) and lowest at E18(25.96±1.31%), and the proportions at E14 and E16 were placed in the middle among the groups.In the cerebral cortex, the highest proportion of NSCs was at E14 (33.65±0.29%), and the lowest at E18(25.29±0.28%), and the middle at E16 (26.82±0.30%).The result of real-time PCR showed that when the mRNA expression of Nestin in the cerebral cortex was set to 1, the relative mRNA expression of Nestin was 0.83±0.04 at E14, 0.77±0.05 at E16, and 0.44 ±0.05 at E18.Thus, the mRNA expression level of Nestin in the mouse cerebral cortex was gradually decreasing with the increase of embryonic days.Conclusions During the brain development, the proportion of NSCs is gradually decreasing in the whole brain and cerebral cortex of mice with the increase of embryonic days.

Objective To investigate the status of the readiness for hospital discharge and pain degree in inpatients with thoracolumbar osteoporotic fractures, and the correlation between them.Methods A cross-sectional analysis of survey data from a sample of 252 patients with thoracolumbar osteoporotic fractures was conducted in a grade A tertiary hospital in Xi′an from January 1st, 2016 to June 30th, 2016. The status of the readiness for hospital discharge and pain level were investigated through the Readiness for Hospital Discharge Scale and Numerical Rating Scale (NRS) respectively. Pearson correlation coefficient method was used to detect the correlation between the two target factors. Results The average score of readiness for hospital discharge was 7.71±1.55. The mean NRS score decreased from 7.8 ± 0.8 at baseline to 2.7 ± 0.6 before discharge. The scores of each dimensions of readiness for hospital discharge from high to low were expected support, personal status, and coping capacity. There was a negative correlation between discharge readiness and pain degree in patients with thoracolumbar osteoporotic fractures (r =-0.537, P<0.05). Conclusions The status of the readiness for hospital discharge among the patients with thoracolumbar osteoporotic fractures is in a medium to high level before discharge. The pain degree is significantly decreased to a lower level. There is a negative correlation between the readiness for hospital discharge and the pain degree.