BACKGROUND: Within the bone marrow stroma there exists a subset of non-hematopoietic stem cells referred to as marrow stromal cells or mesenchymal stem cells. Mesenchymal stem cells (MSCs) are a group of cells with highly capability of self-renew and potential of multilineage differentiation, these properties make them present a promising prospect for clinical practice. Of particular concern is hepatogenic potential that can be used for liver-directed stem cell therapy and transplantation. However, the culture system has not been developed.OBJECTIVE: To explore whether human MSCs are able to differentiate into functional hepatocyte-like cells with hepatocyte growth factor (HGF) and basic fibroblast growth factor (bFGF) in vitro.DESIGN : Open study.SETTING: Department of Infectious Disease and Institute of Urology Surgery, First Affiliated Hospital, Nanchang University.MATERIALS: The study was performed in the Institute of Urolgoy Surgery, the First Affiliated Hospital of Nanchang University from July 2004 to March 2005. Bone marrow was donated by healthy adult with informed consent. DMEM/F12 medium (Gibco); insulin, transferrin, human epidermal growth factor (EGF); human HGF; monoclonal antibodies against human AFP; FITC-conjugated rabbit anti-mouse IgG (Sigma); human bFGF (Invitrogen); monoclonal antibodies against human CK18 and CK19 (Chemicon); fetal bovine serum (Si jiqin, Hangzhou).METHODS: Bone marrow (10 mL) in this study was aspirated from the iliac crest of healthy donors. MSCs were isolated by density gradient centrifugation in combination with plastic adherence. For hepatic differentiation, the 4th- to 8th-passage human MSCs seeded on 24-well tissue culture plates coated with 0.1% gelatin, at 1×104 MSCs/mL, were serum deprived for 2 days, in DMEM/F12 supplemented with 10 μg/L EGF, 10 μg/L bFGF, 5 mg/L insulin and 5 mg/L transferrin. Differentiation was induced by treating MSCs with differentiation medium, consisting of DMEWF12 supplemented with 10 μg/L bFGF, 20 μg/L HGF, 5 mg/L insulin, 5 mg/L transferrin. Medium changes were performed every three days. MSCs without HGF and bFGF in medium served as the control. In the differentiating period, the concentration of AFP in the suernatant was determined dynamically by radioimmunoassay (RIA). The hepatic surface phenotype including AFP, CK18 and CK19 were identified by immunofluorescent staining at day 0, 7, 14, 21 and 28. Glycogen storage was detected by Periodic Acid-Schiff (PAS) staining.MAIN OUTCOME MEASURES: ① the morphological changes of induced MSCs; ② the concentration of AFP in the supernatant; ③ the hepatic surface phenotype; ④ glycogen storage.RESULTS: ① After 14 days ofinduction, the fibroblast-like morphology of human MSCs was lost and cells became broadened and fiattened. After prolonged culture, polygonal cells were seen and further matured hepatocyte-like colonies were seen by day 28. ② The concentration of AFP in the supernatant was first detected on day 14, at a concentration of 0.1 μg/mL, and increased to 0.4 μg/mL by day 17, then decreased to 0.3 μg/mL by day 21. ③ Immunofiuorescent staining showed the expression of AFP and CK18 until day 14. The expression of CK19 was detected by day 28. ④ Glycogen storage could be detected by day 21.CONCLUSION: Human bone marrow MSCs are able to differentiate into functional hepatocyte-like cells and may sere as a new source of cells for cell therapy of hepatic diseases.

Objective To evaluate the effect of oxycodone postconditioning on myocardial ischemia-reperfusion (I/R) injury in rats.Methods Forty pathogen-free healthy adult male Sprague-Dawley rats,weighing 200-300 g,were randomly divided into 4 groups (n =10 each) using a random number table:sham operation group (group S),myocardial I/R group (group Ⅰ),oxycodone postconditioning group (group O),and selective protein kinase C inhibitor chelerythrine group (group CH).Myocardial ischemia was induced by 30 min occlusion of the left anterior descending branch of the coronary artery,followed by 120 min reperfusion.In group S,the left anterior descending branch of the coronary artery was only exposed but not ligated.In group CH,chelerythrine 5 mg/kg was injected intravenously and slowly via the jugular vein before ligation which was performed immediately after administration.In O and CH groups,oxycodone 0.5 mg/kg was injected intravenously and slowly via the jugular vein at 2 min before reperfusion.Arterial blood samples were taken at 120 min of reperfusion to detect the levels of cardiac troponin Ⅰ (cTnI) and creatine kinase-MB (CK-MB) in serum.The hearts were removed after the animals were sacrificed to measure the myocardial infarct size by TTC staining.Results Compared with group S,the levels of cTnI and CK-MB in serum and myocardial infarct size were significantly increased in Ⅰ,O and CH groups (P＜0.05).Compared with group Ⅰ,the levels of cTnI and CK-MB in serum and myocardial infarct size were significantly decreased in O and CH groups (P＜0.05).Compared with group O,the levels of cTnI and CK-MB in serum and myocardial infarct size were significantly increased in group CH (P＜0.05).Conclusion Oxycodone postconditioning can mitigate myocardial I/R injury in rats,and the mechanism is partially related to activation of protein kinase C signaling pathway.

Objective To investigate clinical significance and function of microRNA-106b (miR-106b) in retinoblastoma tissues and cells.Methods We detected miR-106b expression in 51 samples of thyroid cancer and the adjacent non-tumor tissues using qRT-PCR.The expression of miR-106b was altered by corresponding vectors in thyroid cancer cells,and then BrdU cell proliferation and flow cytometry assay were performed to examine the proliferation and apoptosis of thyroid cancer cells.Results The expression of miR-106b in thyroid cancer tissues was significantly lower than that in normal tumor-adjacent tissues (0.36±0.029 vs 0.98±0.034,P＝ 0.004).MiR-106b expression in tumor tissues was significantly associated with tumor size and tumor number.MiR-106b was obviously inhibited by miR-106b inhibitor in PTC-I cells (0.96±0.025 vs 0.29±0.032,P＝0.001),and inhibition of miR-106b resulted in significantly increased proliferation (89.33±5.67 vs 136.67±10.33,P＝0.03) and decreased apoptosis (16.33±3.20 vs 7.67±2.45,P＝0.04).On the contrast,over-expression of miR-106b using miR-106b mimics led to significantly decreased proliferation (98.00±4.65 vs 76.33±2.87,P＝0.03) and increased apoptosis (22.54±2.13 vs 32.45±4.34,P＝0.04).Conclusions Decreased expression of miR-106b is correlated with the adverse clinicopathological features of thyroid cancer.MiR-106b can inhibit cell proliferation and apoptosis of thyroid cancer cells,suggesting miR-106b plays a suppressive role in development and progression of thyroid cancer.

Objective To investigate the effects of sinomenine on hind limb ischemia?reperfusion ( I∕R) injury and expression of Bcl?2 and Bax in skeletal muscle cells of rats. Methods Fifty?four healthy adult male Wistar rats, aged 6-8 weeks, weighing 180-220 g, were divided into 3 groups ( n=18 each) using a random number table: sham operation group ( group S) , group I∕R and sinomenine group ( group SIN) . The rats were subjected to 4 h of ischemia on the proximal part of the right hind limb using elastic rubber bands followed by reperfusion in I∕R and SIN groups. Sinomenine 60 mg∕kg was injected intraperito?neally at 30 min before reperfusion in group SIN, and the equal volume of normal saline was given instead of sinomenine at 30 min before reperfusion in S and I∕R groups. Immediately after onset of reperfusion and at 4 and 24 h of reperfusion, blood samples were collected from the cardiac apex to measure the concentra?tions of serum lactate dehydrogenase ( LDH) and creatine kinase ( CK) . The animals were sacrificed imme?diately after blood sampling, and the gastrocnemius specimens of the hind limb were immediately removed for determination of the wet to dry weight ratio ( W∕D ratio) and expression of Bcl?2 and Bax in gastrocnemi?us cells ( by immunohistochemistry) and for examination of the pathological changes after haematoxylin and eosin staining. The Bcl?2∕Bax ratio was calculated. Results Compared with group S, the gastrocnemius W∕D ratio and concentrations of serum LDH and CK were significantly increased, the expression of Bcl?2 was significantly down?regulated, the expression of Bax was significantly up?regulated, and the Bcl?2∕Bax
ratio was significantly decreased in I∕R and SIN groups ( P<0?05) . Compared with group I∕R, the gastroc?nemius W∕D ratio and concentrations of serum LDH and CK were significantly decreased, the expression of Bcl?2 was significantly up?regulated, the expression of Bax was significantly down?regulated, and the Bcl?2∕Bax ratio was significantly increased in group SIN ( P<0?05) . The pathological changes of the gastrocne?mius were significantly attenuated in group SIN as compared with group I∕R. Conclusion Sinomenine can attenuate hind limb I∕R injury, and the mechanism may be related to maintenance of the balance between Bcl?2 and Bax and to inhibition of apoptosis in skeletal muscle cells of rats.

[ ABSTRACT] AIM:To investigate the role of hydrogen sulfide ( H2 S) on impaired wound healing in ob/ob mice and the underlying mechanism.METHODS:The ob/ob mice were randomly divided into 3 groups, including vehicle, in-sulin and NaHS for treatment.C57BL/6 mice were treated with vehicle as control.Full-thickness punch biopsy wounds were created on the mice.Firstly, H2 S concentrations in the skins and granulation tissues were measured.The mRNA ex-pression of cystathionineγ-lyase ( CSE) was detected by RT-qPCR.The protein expression of CSE and MMP-9 were deter-mined by Western blot.The neutrophil and monocyte/macrophage infiltration was analyzed by immunohistochemistry me-thod.The levels of tumor necrosis factor (TNF)-αand interleukin (IL)-6 were measured by ELISA.Collagen formation was measured by Masson staining.RESULTS:The H2 S levels in the skin and granulation were significantly decreased in ob/ob mice and increased in the NaHS-treated mice ( P<0.05 ) .CSE expression at mRNA and protein levels was significantly decreased in ob/ob mice compared with the control mice (P<0.05).The wound healing period was significantly shorter in NaHS group than that in vehicle-treated ob/ob mice group (P<0.05), in which the insulin group had no difference with vehicle ob/ob mice group.The neutrophil and monocyte/macrophage infiltration, and TNF-αand IL-6 levels were signifi-cantly increased in ob/ob groups, but were decreased in NaHS group ( P<0.01 or P<0.05 ) .Meanwhile, NaHS in-creased collagen formation in the granulation tissues of ob/ob mice.CONCLUSION:H2 S/CSE down-regulation contributes
to impaired wound healing in diabetes, which is alleviated by exogenous H2 S possibly through anti-inflammation.

AIM:To investigate the effect of liraglutide ( LG) on the expression of fibronectin type Ⅲdomain-containing protein 5 (FNDC5) in the C2C12 myotubes.METHODS:The C2C12 mouse myoblast cell line was induced to differentiation.Differentiated cells were stimulated with gradient concentrations (1 ~1000 nmol/L) of LG for different time (0 ~24 h).The effects of LG on the expression of FNDC5 and the activation of adenosine 5'-monophosphate ( AMP)-activated protein kinase ( AMPK) signaling pathway were determined .After pretreated with glucagon-like peptide-1 ( GLP-1 ) receptor antagonist exendin 9-39 , the inhibitor of Ca 2+/calmodulin-dependent protein kinase kinase 2 (CAMKK2), STO609, or the inhibitor of AMPK, Compound C, the LG-induced FNDC5 expression in C2C12 myotubes was examined.The expression of FNDC5 and the activation of AMPK were determined by Western blot .RESULTS: In C2C12 myotubes, LG promoted the expression of FNDC5 in a dose-and time-dependent manner .LG also activated AMPK signaling pathway .These effects of LG were partly abolished by exendin 9-39 , STO609 and Compound C .CONCLUSION:LG promotes the expression of FNDC5 via GLP-1 receptor in the C2C12 myotubes possibly through activation of the CAMKK2/AMPK signaling pathways .

In magnetoencepholography(MEG) inverse research, according to the point source model and distributed source model, the neuromagnetic source reconstruction methods are classified as parametric current dipole localization and nonparametric source imaging (or current density reconstruction). MEG source imaging technique can be formulated as an inherent ill-posed and highly underdetermined linear inverse problem. In order to yield a robust and plausible neural current distribution image, various approaches have been proposed. Among those, the weighted minimum-norm estimation with Tikhonov regularization is a popular technique. The authors present a relatively overall theoretical framework Followed by a discussion of the development, several regularized minimum-norm algorithms have been described in detail, including the depth normalization, low resolution electromagnetic tomography(LORETA), focal underdetermined system solver(FOCUSS), selective minimum-norm(SMN). In addition, some other imaging methods, e.g., maximum entropy method(MEM), the method incorporating other brain functional information such as fMRI data and maximum a posteriori(MAP) method using Markov random field model, are explained as well. From the generalized point of view based on minimum-norm estimation with Tikhonov regularization, all these algorithms are aiming to resolve the tradeoff between fidelity to the measured data and the constraints assumptions about the neural source configuration such as anatomical and physiological information. In conclusion, almost all the source imaging approaches can be consistent with the regularized minimum-norm estimation to some extent.
Algorithms ; Bayes Theorem ; Image Processing, Computer-Assisted ; methods ; Magnetoencephalography ; Models, Statistical ; Nonlinear Dynamics

Objectives Aiming at the current health management mode of chronic disease preven-tion and control as the focus of work, this paper was to evaluate the effect of chronic disease management-based education on clinical teaching and training in cardiovascular medicine. Methods Totally 54 students participating in internship training in Department of Cardiology, the Shanghai Ninth People’s Hospital be-tween July 2013 and Aug 2014 were enrolled. Twenty-eight students were taught by chronic disease man-agement-based method while twenty-six students were taught by traditional method. After the internship training in cardiology, final examination (theoretical exam, skill appraisal and case analysis) were compared between the two groups to assess the teaching efficacy. Questionnaire survey was employed to investigate student satisfaction with chronic disease management-based teaching method. Data were analyzed by SPSS 13.0 software. Quantitative data or enumeration data between two groups were compared by t-test or chi-square test respectively. Results There was no significant difference in the theoretical exam score between two groups (P=0.422). However, the scores of skill appraisal and case analysis in students taught by chronic disease management-based method were significantly higher than those in students taught by traditional method (P<0.05). Most students taught by chronic disease management-based method were satisfied with the novel teaching method. Conclusion In the course of clinical medicine education, the emphasis on the management of chronic diseases can not only significantly improve the quality of clinical teaching in car-diovascular medicine, but also lay the foundation for the implementation of the concept of chronic disease management.

Objective:To investigate the clinical features of Epstein-Barr virus associated lymphoproliferative disease in children and to improve the understanding of this disease.Methods:This study included the children with Epstein-Barr virus associated lymphoproliferative disease admitted to the First Affiliated Hospital of College of Medicine of Zhejiang University from January 2014 to December 2018.Data of these children were collected, including age, clinical manifestations, laboratory results, treatment and outcome.The clinical features and therapeutic effects were analyzed.Results:A total of 114 cases(mean age 6 years, 0~17 years)were enrolled in this study, including 53 males and 61 females.There were 107 cases(93.86%) in the mild group (38 cases of EBV infection and 69 cases of infectious mononucleosis) and 7 cases in the severe group (6.14%). Six cases of the severe group were T cell or NK cell proliferation.Compared with the mild group, the load of EBV-DNA was higher in the severe group, but there was no significant difference( χ2=0.957, P>0.05). The IgM in severe group was significantly lower( Z=-2.041, P<0.05). But the differences in the level of immune function including IgA, IgG, CD4 + cell and CD8 + cell between the severe group and the mild group were not significant.The cases in the mild group had improved after antiviral treatments.Among the severe group, 3 cases survived after treatment, another 1 case was diagnosed as hydroa vacciniforme-like EBV-related proliferative disease (HV-like LPD). After antiviral treatment, the effect was not good, then after high-dose IVIG treatment and Bortezomib combined with methylprednisolone treatment, the EBV-DNA load decreased and the condition improved.While 1 case lost to follow-up, there were 2 cases with EBV-associated hemophagocytic syndrome and 1 case with EBV-associated lymphoma died after chemotherapy or transplantation. Conclusion:EBV-associated lymphoproliferative disease may manifest as a condition similar to infectious mononucleosis.High IgE, low IgM or high DNA load may indicate poor prognosis.Immune function after EBV infection may have different effects on prognosis.When the infected lymphocyte types are NK or T cells, it may indicate poor prognosis.The efficacy of transplantation and chemotherapy in severe cases is still uncertain.

Objective To evaluate the effect of pre-cut-endoscopic mucosa resection of colorectal laterally spreading tumor. Methods 65 patients with LST were enrolled from January 2014 to February 2014. LST was detected by chromoendoscopy and NBI combined with magnifying endoscopy technique. The size, site, morphological features, were observed and the histopathological features of the specimen of LST was analyzed. All the 65 LSTs were resect by pre-cut-EMR. The clinical results including enbloc resection rate, all bloc resection rate, procedure time, complication and recurrence rates were retrospectively evaluated. Results All the 65 LSTs lesions ranged from 2.0 cm to 5.0 cm, with a mean diameter of (2.4 ± 1.7) cm. The site of 65 LSTs was in rectum 28 (43.1 %), 11 LSTs in sigmoid colon (16.9 %), 6 LSTs in descending colon (9.2 %), 2 LSTs in splenic flexure of colon (3.1 %), 9 LSTs in transverse colon (13.8 %), 4 LSTs in Hepatic flexure of colon (6.2 %), 2 LSTs in ascending colon(3.1 %), and 3 LSTs in cecum (4.6 %). Morphology of 23 LSTs were homogeneous granular type (35.4 %), 27 LSTs were mixed non-granular type (41.5 %), 13 LSTs were flat elevated type (20.0 %), and 2 LSTs were pseudo-depressed type (3.1 %). The histopathological diagnoses of LST included 12 tubular adenoma (18.5 %), 19 villous-tubular adenoma (29.2 %), 26 villous adenoma (40.0 %), 7 advanced intraepithelial tumor (10.7 %), 1 intramucosal carcinoma (1.5 %). Enbloc resection was achieved in 65 patients (100.0 %) with a mean operation time of (18.0 ± 11.7) min. 5 cases were bleeding during the operation (7.7 %), 1 case was bleeding 7 days after operation (1.5 %), no perforation was happened. 65 patients were followed up for 3 ~ 12 months, and no local recurrence was found. Conclusion Pre-cut-endoscopic mucosal resection an effective and safe therapy for colorectal LST larger than 2.0 cm.