Objective To investigate the value of the E-selectin,IL-6 and TNF-α for the diagnosis of myocardial damage in children.Methods The E-selectin,IL-6 and TNF-α levels were detected in 62 patients with myocardial damage and 62 children without myocardial damage.The values of the E-selectin,IL-6 and TNF-α for the diagnosis of myocardial damage were analyzed by the receiver operating characteristic (ROC) curve.Results E-selectin levels were (45.16 ± 5.34) and (15.37 ± 4.65) μg/L,respectively in the patients with myocardial damage and the control group,and the difference between the two groups was significant(P ＜ 0.001).IL-6 levels were (21.31 ± 5.28) ng/L and (17.09 ± 4.62) ng/L,TNF-α levels were (27.22 ± 8.56) ng/L and(21.46 ± 8.08) ng/L,showing no significant difference (P ＞ 0.05).ROC curve analysis showed that AOC of E-selectin in the diagnosis of myocardial damage was 0.945 with 95％ CI 0.899 ～0.991 (sensitivity:0.801,specificity:0.999,Youden Index:0.800),and the cutoff value was 29.93 μg/L.Conclusion E-selectin can be used as one of the diagnostic indices for myocardial damage in children,which has great diagnostic value.IL-6 and TNF-α are of limited value in the diagnosis of myocardial damage.

Background and purpose:The expression of ERβin triple negative breast cancer(TNBC) might be associated with good prognosis in TNBC patients. ERβand ERαhave considerable homology. FOXA1 plays an important role in ERαexpression and function. The aim of this study was to analyze the expression of FOXA1 and ERβin TNBC and the relationship between them and the clinicopathologic characteristics and prognosis. Methods:The breast cancer samples in Peking Union Medical College Hospital were collected from Nov. in 2011 to Dec. in 2013, and TNBC were screened out based on the expression of ERα, PR and HER-2. Thirty ERβ-negative samples and 30 ERβ-positive samples were selected randomly according to the ERβexpression. We used immunohistochemical method to detect the expression of FOXA1. Finally, 48 TNBC samples were obtained to analyze the results. Results:The total positive rate of FOXA1 was 35.4%(17/48). In the ERβ-positive group, the positive rate of FOXA1 was 35.7%(10/28),and in the ERβ-negative group, the positive rate of FOXA1 was 35% (7/20). The expression of FOXA1 in these 2 groups had no signiifcant difference (P=0.83), which indicated that there was no relation between ERβand FOXA1. The FOXA1 positive group and FOXA1 negative group also showed no signiifcant difference in age, tumor size, and lymphatic metastasis number in axilla, tumor grade, tumor stage, NPI and DFS. However, Ki-67 showed negative correlation with FOXA1 expression (P<0.01). Conclusion:FOXA1 expression had no relationship with ERβexpression in TNBC. Ki-67 showed negative correlation with FOXA1 expression, which might hint that the proliferation of tumor cell was lower in FOXA1 positive TNBC.

Aim To evaluate the efficacy of TFU as a precursor of 5-FU on the growth inhibition of human gastric carcinoma cell lines SGC-7901 and MKN-45.Methods In vitro experiments,cell growth inhibition was measured by MTT assay.The rates were compared in the presence and Absence of liver microsomal enzymes.The morphology of apoptotic cells was detected by observation with a fluorescence microscope after staining by using adridine orange/ethidium bromide solution.DNA fragmentation was analyzed by agarose gel electrophoresis and flow cytometry respectively.Western blot was employed to analyze the expression of Bcl-2 and Bax.The in vivo efficacy of TFU was assessed in nude mice bearing tumours.The specimens were re-moved and the in situ cell apoptosis detection kit was employed for TUNEL staining.Results Growth of SGC-7901 and MKN-45 cells was remarkably suppressed by treatment with TFU in the presence of liver microsomal enzymes in vitro,suggesting that TFU might be converted to 5-FU by the enzymes.Similar treatment of TFU induced apoptosis of the cells,which was deduced from typical apoptotic features such as morphology,the formation of characteristic ladder pattern of DNA migration and the accumulation of sub-G1 phase.Furthermore,a significant inhibition of Bcl-2 expression and the up-regulation of Bax were observed after treatment with TFU in the presence of liver microsomal enzymes.Growth of human gastric carcinoma cells was significantly delayed by oral administration of TFU with low side effects.Apoptosis in xenografts was also observed by means of TUNEL staining method.Conclusion Treatment of TFU in the presence of liver microsomal enzymes could promote the inhibition of gastric carcinoma cell proliferation.TFU might sustain release of 5-FU mediated by liver microsomal enzymes.Low dose of 5-FU might trigger the carcinoma cells apoptosis via regulation of Bax and Bcl-2.

Aim To investigate the protective effects of LMWH-SOD(Low molecular weight heparin- Superoxide dismutase Conjugate)on the injuries induced by oxygen and glucose deprivation in cultured neurons. Methods The cortical neurons of fetal rat were cultured in vitro. The antioxidant and protective effects of LMWH-SOD were observed by treating neurons with oxygen and glucose deprivation. Results LMWH-SOD reduced the number of cell death and the efflux of LDH and the content of NO,MDA and increased the membrane fluidity after the injuries of cells. Conclusion LMWH-SOD has protective effects on cerebral cortical neurons through its action of scavenging free radicals.

Objective: To investigate the early effects of acellular xenogeneic nerve combined with adipose-derived stem cells (ADSCs) and platelet rich plasma (PRP) in repairing facial nerve injury in rabbits. Methods: The bilateral sciatic nerves of 15 3-month-old male Sprague-Dawley rats were harvested and decellularized as xenografts. The allogeneic ADSCs were extracted from the neck and back fat pad of healthy adult New Zealand rabbits with a method of digestion by collagenase type Ⅰ and the autologous PRP was prepared by two step centrifugation. The 3rd generation ADSCs with good growth were labelled with CM-Dil living cell stain, and the labelling and fluorescence attenuation of the cells were observed by fluorescence microscope. Another 32 New Zealand rabbits were randomly divided into 4 groups and established the left facial nerve defect in length of 1 cm ( n=8). The nerve defects of groups A, B, C, and D were repaired with CM-Dil-ADSCs composite xenogeneic nerve+autologous PRP, CM-Dil-ADSCs composite xenogeneic nerve, xenogeneic nerve, and autologous nerve, respectively. At 1 and 8 weeks after operation, the angle between the upper lip and the median line of the face (angle θ) was measured. At 4 and 8 weeks after operation, the nerve conduction velocity was recorded by electrophysiological examination. At 8 weeks after operation, the CM-Dil-ADSCs at the distal and proximal ends of regenerative nerve graft segment in groups A and B were observed by fluorescence microscopy; after toluidine blue staining, the number of myelinated nerve fibers in regenerated nerve was calculated; the structure of regenerated nerve fibers was observed by transmission electron microscope. Results: ADSCs labelled by CM-Dil showed that the labelling rate of cells was more than 90% under fluorescence microscope, and the labelled cells proliferated well, and the fluorescence attenuated slightly after passage. All the animals survived after operation, the incision healed well and no infection occurred. At 1 week after operation, all the animals in each group had different degrees of dysfunction. The angle θ of the left side in groups A, B, C, and D were (53.4±2.5), (54.0±2.6), (53.7±2.4), and (53.0±2.1)°, respectively; showing significant differences when compared with the healthy sides ( P<0.05). At 8 weeks after operation, the angle θ of the left side in groups A, B, C, and D were (61.9±4.7), (56.8±4.2), (54.6±3.8), and (63.8±5.8)°, respectively; showing significant differences when compared with the healthy sides and with the values at 1 week ( P<0.05). Gross observation showed that the integrity and continuity of regenerated nerve in 4 groups were good, and no neuroma and obvious enlargement was found. At 4 and 8 weeks after operation, the electrophysiological examination results showed that the nerve conduction velocity was significantly faster in groups A and D than in groups B and C ( P<0.05), and in group B than in group C ( P<0.05); no significant difference was found between groups A and D ( P>0.05). At 8 weeks after operation, the fluorescence microscopy observation showed a large number of CM-Dil-ADSCs passing through the distal and proximal transplants in group A, and relatively few cells passing in group B. Toluidine blue staining showed that the density of myelinated nerve fibers in groups A and D were significantly higher than those in groups B and C ( P<0.05), and in group B than in group C ( P<0.05); no significant difference was found between groups A and D ( P>0.05). Transmission electron microscope observation showed that the myelinated nerve sheath in group D was large in diameter and thickness in wall. The morphology of myelin sheath in group A was irregular and smaller than that in group D, and there was no significant difference between groups B and C. Conclusion: ADSCs can survive as a seed cell in vivo, and can be differentiated into Schwann-like cells under PRP induction. It can achieve better results when combined with acellular xenogeneic nerve to repair peripheral nerve injury in rabbits.

Objective To evaluate the effect of exogeneous adiponectin on hippocampal advanced glycation end products (AGEs)-reactive oxygen species (ROS)-endoplasmic reticulum stress (ERS) pathway in aged mice with postoperative cognitive dysfunction (POCD).Methods Thirty-two healthy male C57BL/6 mice, aged 18 months, weighing 20-25 g, were randomly divided into 4 groups (n=8 each) using a random number table: control group (group C), POCD group, exogeneous adiponectin group (group APN), and vehicle group (group Veh).Splenectomy was performed to establish the POCD model in aged mice anesthetized with intraperitoneal pentobarbital sodium.In group APN, adiponectin 0.1 μg/g (in 2 μl of phosphate buffer solution) was injected into the lateral cerebral ventricle at 30 min before establishing the model.Phosphate buffer solution 2 μl was given at 30 min before establishing the model in group Veh.Cognitive function was assessed on day 7 after surgery.The mice were then sacrificed, and the hippocampus was harvested for determination of the area of AGE deposition (by immunohistochemistry), levels of ROS (by flow cytometry), and levels of glucose-regulated protein 78 (GRP78), C/EBP-homologous protein (CHOP), caspase-12 and ROS (using Western blot).Results Compared with group S, the freezing time in the contextual fear conditioning test was significantly shortened, the area of AGE deposition and levels of ROS, CHOP and caspase-12 were increased, and the level of GRP78 was decreased in POCD, APN and Veh groups.Compared with POCD and Veh groups, the freezing time in the contextual fear conditioning test was significantly prolonged, the area of AGE deposition and levels of ROS, CHOP and caspase-12 were decreased, and the level of GRP78 was increased in group APN.Conclusion Exogeneous adiponectin decreases the occurrence of POCD probably by blocking hippocampal AGEs-ROS-ERS pathway in aged mice.

Objective To investigate the role of hippocampal cyclophilin D (CypD) in sepsis-associated encephalopathy in rats.Methods A total of 36 adult male Sprague-Dawley rats,aged 3-4 months,weighing 300-400 g,were randomly divided into 3 groups (n =12 each) using a random number table:sham operation group (Sham group),sepsis group (S group),and sepsis + CypD inhibitor cyclosporin A group (CsA group).Sepsis was induced by cecal ligation and puncture (CLP).Cyclosporin A 6 mg/kg was injected intraperitoneally at 30 min before CLP in group CsA.All the animals underwent Morris water maze test on 4th day after CLP.The animals were sacrificed after the test,and the hippocampus was isolated for determination of the expression of cytochrome c (Cyt c),CypD,caspase-3,brain-derived neurotrophic factor (BDNF),phosphorylated protein kinase A (p-PKA),and phosphorylated cyclic adenosine monophosphate response element-binding protein (p-CREB).Results Compared with group Sham,the escape latency was significantly prolonged,the space exploration time was shortened,the expression of Cyt c,CypD,caspase-3,p-PKA and p-CREB was up-regulated,and the expression of BDNF was down-regulated in S and CsA groups (P＜0.05).Compared with group S,the escape latency was significantly shortened,the space exploration time was prolonged,the expression of Cyt c,CypD,caspase-3,p-PKA and p-CREB was down-regulated,and the expression of BDNF was up-regulated in group CsA (P＜0.05).Conclusion Hippocampal CypD may be involved in the pathophysiological mechanism of sepsis-associated encephalopathy,and the downstream mechanism is probably related to promotion of activation of PKA/CREB signaling pathway in rats.

Objective To investigate the effects of acetylated HMGB1 on cognitive function in rats with sepsis associated encephalopathy (SAE) and the effect of HMGB1 inhibitor.Methods Forty-eight males mice were randomly assigned to three groups (n=16): sham group (group S),cecal ligation puncture group (group C),cecal ligation puncture+sodium butyrate group (group B).Cecal ligation puncture was applied to establish the SAE model,and group S received sham operation.Rats in groups S and C were injected with normal saline 5 ml/kg 30 min and 4 h after CLP,respectively.The rats in group B were intraperitoneally injected with sodium butyrate 500 mg/kg 0.5 h and 4 h after CLP,respectively.All animals were performed Morris water maze test on 4th day after operation,and the exploring time of space exploration experiments were assessed on 7th day after CLP surgery.IL-6,BDNF,HMGB1 and acetylated HMGB1 expression in hippocampus of all rats were determined by Western Blot.Results Compared with group S,the latency of rats in group C was longer and the exploring time was shorter (P<0.05).Compared with group C,the latency of rats in group B was shorter and the exploring time was longer (P<0.05).Compared with group S,the expression of IL-6,HMGB1 and acetylated HMGB1 in group C increased (P<0.05) and the level of BDNF decreased (P<0.05).Compared with group C,the expression of IL-6,HMGB1 and acetylated HMGB1 in group B decreased (P<0.05) and the level of BDNF increased (P<0.05).Conclusion HMGB1 inhibitor sodium butyrate can inhibit the expression of acetylated HMGB1 in the hippocampus of SAE rats,and reduce the cognitive impairment induced by sepsis.

Objective To explore the association of estrogen receptor β expression with different stages and molecular subtypes of invasive breast cancer.Methods The clinicopathologic data of 446 invasive breast cancer cases was retrospectively analyzed.ERβ expression was evaluated by types and stages.Results Of all 446 invasive breast cancer cases,328 (73.5％) were ERβ positive.The ERβ positive rate was 77.9％ (240/308) and 63.8％ (88/138) in ERα + group and ERα-group,respectively.The ERβ expression in breast cancer was positively correlated with ERα (P ＜ 0.01) while it had no correlation with PR,histological grade,HER-2 and Ki-67 (P ＞ 0.05).ERβ expression was not significantly different among different age,tumor size and axillary lymph node groups(all P ＞ 0.05).A total of 418 invasive breast cancer cases were recruited for pathologic stage and NPI analysis,including 168 cases at stage Ⅰ,152 cases at stage Ⅱ and 98 cases at stage Ⅲ.ERβ expression was not significantly different among different stages of breast cancer(P =0.743).Analyzed in these 418 cases,NPI was ＜ 3.4 in 126 cases,3.4-5.4 in 207 cases and ＞ 5.4 in 85 cases.ERβ expression was not significantly different among different NPI group (P =0.644).The ERβ positive rate in Luminal A subtype,Luminal B1 subtype,Luminal B2 subtype,HER-2 subtype and TN subtype was 75.6％ (88/118),75.9％ (110/145),85.2％ (46/54),68.4％ (39/57) and 62.5％ (45/72) respectively.ERβ expression was significantly different between Luminal subtype and non-Luminal subtype (P =0.007).Conclusions ERβ was not differentially expressed among different breast cancer stages and NPI groups.ERβ was differentially expressed in different breast cancer molecular subtypes.