BACKGROUND: Studies regarding Feridex in vitro cell labeling are mainly in rodents, while little information is known on primate crab-eating macaque.OBJECTIVE: To explore the feasibility of protocols using Feridex and transfection agents for in vitro magnetic labeling of bone marrow stromal cells (BMSCs) in crab-eating macaque.METHODS: Under the sterile condition, the crab-eating macaque BMSCs were obtained by means of density gradient centrifugation following a bone puncture. Feridex-poly-l-lysine complexes were used to magnetically label BMSCs. The efficiency and cellular viability of Feridex-poly-l-lysine labeled BMSCs were evaluated by Prussian blue staining, electron microscopy, and trypan blue dye exclusion test. The proliferation and differentiation ability of Feridex-poly-l-lysine labeling BMSCs were also investigated by inverted phase contrast microscope and immunocytochemistry. RESULTS AND CONCLUSION: BMSCs could be effectively labeled by Feridex and labeling efficiency was around 99%. Tiny blue stained fine particles and numerous vesicles coated with the electron-dense magnetic iron particles could be found in the cytoplasm of Feridex-poly-l-lysine labeled BMSCs under optical microscopy and transmission electron microscopy respectively. Cell viability, proliferation and differentiation ability of labeled BMSCs were not affected by Feridex-poly-l-lysine labeling. Results indicated that Feridex might be used to label BMSCs of crab-eating macaque.

Objective To study the mediated neuroprotective effect of B-cell lymphoma/Ieukemia -2 ( Bcl -2 ) and hairy enhancer of split homologues 1 ( Hes -1 ) after cerebral ischemic preconditioning.Methods The model A of cerebral ischemia was set up by the way of middle cerebral artery occlusion.The right internal carotid artery of rat was blocked transiently 10 minutes at a time, then after 3 days the model B was established by the way of model A.The sham operation group was only isolated carotid artery.The time 2, 3, 7, 14 days after the proce-dure , the neurobehavioral function in rat was evaluated by the way of Zea-Longa.The volume of cerebral infarction was measured by 2 , 3 , 5-triphenyltetrazolium chloride ( TTC) stain.The mRNA expression of Bcl-2 and Hes1 were assessed by real-time fluorescence quantitative polymerase chain reaction ( qRT -PCR ) .Results Compared with model A group, the neurologic impairment scores and cerebral infarction volume[ A:( 40.00 ±4.47 )%, B:( 19.00 ±2.75 )%, respectively ] of rats in model B group had obvious discrepancy( P<0.05).Hes1 mRNA expression in model A and model B were higher than sham operation group, model B were higher than model A, the difference had significantly (P<0.05).Bcl-2 mRNA expression, model B were significantly higher than sham group and model A at 2 days, the difference had significantly( P<0.05).Conclusion The Bcl-2 and Hes1 mRNA ex-pression increased after cerebral ischemia precondition, may be associated with nerve protective effect.

AIMTo observe the role of melatonin receptor and GABAA receptor in sleeping time prolonged by melatonin in mice.

METHODSThe absence of the righting reflex was considered as the sleep onset and the duration of the loss of the righting reflex was recorded as the sleeping time. The effects of receptor agonist and antagonist on hypnotic activity of melatonin were studied in the paper.

RESULTSPrazosin hydrochloride, the blocker of melatonin 3 receptor, didn't affect the sleeping time prolonged by melatonin in mice. GABA, the endogenous agonist of GABA receptor, significantly potentiated the hypnotic activity of melatonin. When picrotoxin, the ligand of picrotoxin site on GABAA receptor, used together with melatonin, it significantly antagonized the sleeping time prolonged by melatonin, however, bicuculline, the specific antagonist of GABA binding site in GABAA receptor, didn't affect the hypnotic activity of melatonin in mice.

CONCLUSIONMelatonin does not exhibit its potentiation sleeping time in mice through melatonin 3 receptor. Hypnotic activity of melatonin may be mediated through picrotoxin site on GABAA receptor.

Animals ; Bicuculline ; pharmacology ; Male ; Melatonin ; physiology ; Mice ; Mice, Inbred Strains ; Picrotoxin ; pharmacology ; Prazosin ; pharmacology ; Receptors, GABA-A ; physiology ; Receptors, Melatonin ; physiology ; Sleep ; physiology

OBJECTIVETo study the effect of integrin alpha2beta1 on invasion and migration of SK-N-SH neuroblastoma cells.

METHODSNeuroblastoma SK-N-SH cell line was cultured in the modified eagle's medium. The effects of monoclonal antibodies to integrin alpha2 and integrin beta1 on migration and invasion were measured by inclined test and polycarbonate filters incorporated in modified Transwell chambers respectively. The migration and invasion cells were stained with Gimsa staining and counted under a 200 multiplied microscope. The blocking rate of migration and invasion of cells was calculated.

RESULTSThe number of migrated SK-N-SH cells in the anti-alpha2 and anti-beta1 treatment groups (50.9+/-10.5 and 54.3+/-9.0 respectively) was significantly less than that in the control group without monoclonal antibody treatment (98.1+/-7.4) (P<0.01), with a blocking rate of cell migration of 48.1% and 44.5% respectively. The invasion to matrigel of SK-N-SH cells exposed monoclonal antibodies to integrin alpha2 and integrin beta1 was significantly blocked compared with the control SK-N-SH cells, with the number of invasion cells in the anti-alpha2 and anti-beta1 treatment groups of 25.3 +/- 4.4 and 18.8 +/- 3.9 respectively vs 41.5 +/- 4.8 in the control group (P<0.01). The blocking rate of cell invasion in the anti-alpha2 and anti-beta1 treatment groups was 39.0% and 54.7% respectively.

CONCLUSIONSIntegrin alpha2beta1 may promote migration and invasion of neuroblastoma cells.

Cell Line, Tumor ; Cell Movement ; Collagen Type I ; physiology ; Humans ; Integrin alpha2beta1 ; physiology ; Neoplasm Invasiveness ; Neuroblastoma ; pathology

Considering the existing shortages in evaluation criteria, training systems and management modes, administrators from graduate schools of Shanghai Jiao Tong University School of Medicine (SJTUSM) performed reforms and practice for the establishment of tutor teams which should meet the requirements for culturing talented medical graduates in recent years. We have established a multidimensional evaluation criteria system oriented by both representative academic achievements and the quality of graduate students. We have strengthened horizontal cooperation with relevant administrative departments in SJTUSM. We also emphasize the concepts of "training the trainees" coordinated with administrators in the training units to implement reform measures. Herein we have summarized the main reform measures in SJTUSM to constantly improve the quality of postgraduate tutor team and guarantee high-level postgraduate education.

Objective To investigate the expression changes of the hydrogen sulfide synthases cystathionineγ-lyase (CSE), cystathionineβ-synthase (CBS), and 3-mercaptopyruvate sulfurtransferase (3-MST), after optic nerve crush (ONC) in rat the retina.Methods The rat model of ONC was established.Rats were divided into normal control, ONC, and sham control groups.Histopathologic changes in retina, the number of retinal ganglion cells (RGC) and retinal thickness of inner part (RTIP) were measured.The changes of CSE, CBS and 3-MST mRNA expression were detected with quantitative real-time PCR.Results The retinal histostructure was normal in normal controls and with minor changes in sham controls, respectively.Compared with sham group, significant retina damages were found in the ONC group:a time-dependent reduction of RGC number and RTIP.Expressions of CSE, CBS and 3-MST mRNA in rat retina were detected in normal control.Compared with normal controls, the expressions of CSE, CBS and 3-MST mRNA did not show any significant changes in the sham controls.Compared with sham controls, CBS mRNA expressions showed a time-dependent increase at 3 d, 7 d and 14 d after crush in the ONC group;CSE mRNA expressions increased to the peak at 3 d and then slightly reduced at 14 d after crush;3-MST mRNA expressions showed the trend of increase at 3 d and 7 d and then enhanced remarkably at 14 d after crush.Conclusion Hydrogen sulfide synthases CSE, CBS and3-MST expressions were up-regulated in rat retina following ONC, which may cause an increase in local endogenous hydrogen sulfide production in the retina and a compensatory protective effect.

Objective:To investigate the role of umbilical cord mesenchymal stem cell-derived exosomes (ucMSC-exos) in acute skin wound healing in mice.Methods:ucMSC-exos were extracted by ultracentrifugation, and identified by transmission electron microscopy, Western blot analysis of exosome surface markers CD63 and TSG101, and particle size analysis. Firstly, in vitro cultured third- to fifth-passage human skin fibroblasts (HSF) were incubated with high-glucose Dulbecco′s modified Eagle′s medium (DMEM) containing 0, 1 and 2 μg/ml exosome suspension for 24 hours (negative control group, 1- and 2-μg/ml groups, respectively) , and cell counting kit-8 (CCK8) assay was performed to evaluate the effect of ucMSC-exos on the proliferative activity of HSF. Secondly, 24 male BALB/c mice aged 8 weeks were selected to construct a mouse model of full-thickness skin wound, and then divided into ucMSC-exos group and phosphate-buffered saline (PBS) group by using a random number table to be subcutaneously injected with exosome suspension and PBS respectively at multiple equidistant sites located about 1 mm apart from the wound edge. On days 0, 4, 7, 10 and 14 after operation, the wounds in mice were observed, and the percentage of residual wound area was calculated in the above two groups. On days 7 and 14 after operation, wound tissues were resected and subjected to hematoxylin and eosin (HE) and Masson staining to observe structural changes of skin tissues. On day 14 after operation, wound tissues were collected in the two groups, and real-time quantitative PCR (qRT-PCR) and Western blot analysis were performed to determine the mRNA and protein expression of type Ⅰ collagen, fibronectin and vascular endothelial growth factor, respectively. Statistical analysis was carried out by using one-way analysis of variance, least significant difference- t test, two-way repeated measures analysis of variance and unpaired t-test. Results:Under the transmission electron microscope, the ucMSC-exos were oval in shape with a diameter of about 100 nm; Western blot analysis showed positive expression of ucMSC-exos surface proteins CD63 and TSG101; particle size analysis showed that 96.2 % of the ucMSC-exos had diameters of 30 - 150 nm. CCK8 assay showed that the relative proliferative activity of HSF was significantly higher in the 1- and 2-μg/ml groups (0.97 ± 0.05, 1.08 ± 0.07, respectively) than in the negative control group (0.71 ± 0.04; t = 2.00, 7.05, respectively, both P < 0.05) , and significantly higher in the 2-μg/ml group than in the 1-μg/ml group ( t = 5.09, P < 0.05) . On days 4, 7, 10 and 14 after operation, the percentage of residual wound area was significantly lower in the ucMSC-exos group than in the PBS group (all P < 0.05) . HE and Masson staining showed increased numbers of hair follicles, glands and granulation tissues, more neovascularization, and neater arrangement of collagens in neonatal skin tissues of the mice in the ucMSC-exos group compared with the PBS group. qRT-PCR and Western blot analysis showed significantly increased mRNA and protein expression of type Ⅰ collagen, fibronectin and vascular endothelial growth factor in the ucMSC-exos group compared with the PBS group (all P < 0.01) . Conclusion:Subcutaneous injections of ucMSC-exos can promote acute skin wound healing in mice, likely by promoting the synthesis of extracellular matrix and vascular endothelial growth factor in wound tissues of mice and proliferation of HSF.

Objective To investigate the expression of amyloid precursor protein (APP), Fis1, PDCD7 and KLRF1 mRNA in acute myeloid leukemia (AML) and their prognostic significances. Methods Thirty-four AML patients and 10 patients with nonmalignant hematologic diseases (control group) were recruited in this study. Bone marrows were obtained from these patients to isolate the mononuclear cells, from which total RNA was extracted and reverse transcribed into cDNA. SYBR Green Ⅰ dye-based real-time quantitative PCR method was used to compare the expression of APP, Fis1, PDCD7 and KLRF1 mRNA between two groups. Results The expression level of PDCD7 mRNA in AML patients was significantly higher than that in the control group (Z=-2.213,P=0.027), but APP, Fisl and KLRF1 mRNA expression levels were similar between the two groups (P<0.05). The expression level of APP mRNA in M2 patients with t (8, 21) chromosomal abnormality was significantly higher than that in the control group (Z=-2.197, P=0.028), while APP mRNA expressions in M4b and M5b patients were both lower than those in the control group (Z=-2.368, P-0.018). The expression of APP mRNA differed significantly between the subtypes of AML, higher in M2 than in M4b and M5b (Z=-2.430, P=0.015; Z=-3.175, P=0.001, respectively). Conclusions The expression of PDCD7 mRNA increased significantly in AML patients, and APP mRNA expression is decreased in monocytic leukemia.

Objective To investigate the expression of amyloid precursor protein (APP), Fis1, PDCD7 and KLRF1 mRNA in acute myeloid leukemia (AML) and their prognostic significances. Methods Thirty-four AML patients and 10 patients with nonmalignant hematologic diseases (control group) were recruited in this study. Bone marrows were obtained from these patients to isolate the mononuclear cells, from which total RNA was extracted and reverse transcribed into cDNA. SYBR Green Ⅰ dye-based real-time quantitative PCR method was used to compare the expression of APP, Fis1, PDCD7 and KLRF1 mRNA between two groups. Results The expression level of PDCD7 mRNA in AML patients was significantly higher than that in the control group (Z=-2.213,P=0.027), but APP, Fisl and KLRF1 mRNA expression levels were similar between the two groups (P<0.05). The expression level of APP mRNA in M2 patients with t (8, 21) chromosomal abnormality was significantly higher than that in the control group (Z=-2.197, P=0.028), while APP mRNA expressions in M4b and M5b patients were both lower than those in the control group (Z=-2.368, P-0.018). The expression of APP mRNA differed significantly between the subtypes of AML, higher in M2 than in M4b and M5b (Z=-2.430, P=0.015; Z=-3.175, P=0.001, respectively). Conclusions The expression of PDCD7 mRNA increased significantly in AML patients, and APP mRNA expression is decreased in monocytic leukemia.

OBJECTIVETo investigate the prevalence, etiology and clinical characteristics of adrenal lesions detected by abdominal computed tomography (CT).

METHODSThis retrospective study was conducted in patients with adrenal lesions detected by abdominal CT examinations in Nanfang Hospital between July, 2014 and June, 2015. The clinical data of the patients were collected for analysis of the demographics, comorbidities, imaging characteristics, biochemical profiles, clinical diagnosis and intervention.

RESULTSA total of 939 patients with adrenal lesions were identified from 19 004 patients undergoing abdominal CT scan over the defined period. The mean age of the patients was 53.2 years and 560 of the patients were male. Among the total cases with adrenal lesions, the percentages of cases with adrenal masses tended to increase progressively with age. Endocrine studies were done in 270 of the total patients, which identified non-functioning masses in 38.9%, primary aldosteronism in 16.3%, Cushing's syndrome in 4.1%, subclinical Cushing's syndrome in 7.0%, and pheochromocytomas in 7.0% of the cases. Adrenal incidentalomas was detected in 191 patients, with a detection rate of 1.0% among the overall patients undergoing abdominal CT scans. Imaging study detected adenomas (70.3%), cortical carcinomas (2.4%), and metastases (0.5%). Of 191 patients with adrenal incidentalomas, only 76 (39.8%) underwent endocrine evaluation, including 34 with nonfunctioning adrenal masses, 17 with pheochromocytoma, 7 with primary aldosteronism, and 5 with subclinical Cushing's syndrome.

CONCLUSIONs The overall detection rates of adrenal lesions and adrenal incidentalomas by abdominal CT were 4.9% and 1.0%, respectively, in our cohort of patients undergoing the examination over the defined period. Although most of the lesions were benign and nonfunctioning, malignant and functional lesions were also detected. As many as 60% of the patients with adrenal incidentalomas did not have hormonal testing. Clinicians need to have greater awareness of adrenal incidentalomas and standard protocol for its management should be established.