Aim To observe the effect of sodium nitrop russide (SNP)on glial fibrillary acidic protein(GFAP) synthesis in the gerbil hippocampus. Method lmmunofluorescent histochemical staining method was used. Result SNP increased GFAP synthesis in rediatum layer,molecular layer and dentate gyrus.There were not GFAP positive cells in rediatum layer and mol ecular layer.Number of GFAP positive cells related to dose of SNP.Conclu sion SNP increased GFAP synthesis.

Objective To explore the role of ERK1/2 in the central pathogenesis of migraine.Methods Healthy adult male SD rats were randomly divided into five groups:normal group (group C),sham operation group(group C),migraine model group(group M),DMSO group (group D)and PD-98059group (PD group),with 12 rats in each group.The extracellular discharge frequency in the spinal trigeminal nucleus was recorded and ERK1/2 phosphorylation was tested.Results (1) The percentage of extracellular discharge frequency change:Two hours after treatment,the percentage of discharge frequency change was (325.9±47.32)％.The percentage of extracellula discharge frequency change in group M (325.9±47.3)％ was higher than that in group N (100.0± 0.0) ％ and group C(107.3± 16.4)％.There was no significant difference in the percentage of discharge frequency change between group D(319.3±42.5) ％ and group M (325.9±47.3) ％.The percentage of discharge frequency change in group PD(218.5±31.7)％ was lower than that in group M(325.9±47.3)％ and group D(319.3± 42.5)％.(2) ERK1/2 phosphorylation:the ERK1/2 phosphorylation in group M and group D was higher than that in group N and group C.There was no significant difference in ERK1/2 phosphorylation between group D and group M.The ERK1/2 phosphorylation in group PD was lower than the other four groups.Conclusion During the process of central sensitization to migraine,neuronal excitability and ERK1/2 phosphorylation were increased.ERK1/2 inhibitor (PD98059) reduced ERK1/2 phosphorylation and neuronal excitability.These indicated that ERK1/2 may play a role in central sensitization of migraine in rats.

BACKGROUND: Nanotechnology has widely used in tissue engineered reconstruction in recent years. Most reports are concerning carbon nanomaterials in bone reparation, but the study of peripheral nerve regeneration is poorly understood.OBJECTIVE: To improve the physical, chemical and biological properties of chitosan/collagen composite nerve conduit with functionalized carbon nanotubes, in addition, to investigate the therapeutic effect of this novel material.DESIGN, TIME AND SETTING: The same body controlled experiment of animals was performed at the Tissue Engineering Laboratory and The Key Laboratory of Thin Film and Microfabrication Technology, Shanghai Jiao Tong University from February 2005 to November 2006.MATERIALS: The carbon nanotubes were mixed with 2% chitosan solution, coated on the die to prepare chitosan/collagen composite nerve conduit with functionalized carbon nanotubes. The chitosan/collagen tubes were served as controls.METHODS: A total of 80 male adult-rats were prepared a 4 mm accessory nerve defects models, and repaired by nerve conduit in the experimental material and control material groups. In the auto nerve grafts group, the removed nerve was connected to the broken end. In the blank control group, there was no other treatment except removing 2 mm nerves. The left sides were served as experimental sides and the right sides as within-subject controls.MAIN OUTCOME MEASURES: The repairing outcomes were measured by electrophysiological, myophysiological, and histological measurements.RESULTS: The accessory nerve defects were repaired in a rat model using carbon nanotubes in chitosan/collagen-based composite nerve conduit. As time passed after the surgery, good results of the electrophysiological, myophysiological and histological measurements were achieved, which were similar or superior to those of the nerve autografts.CONCLUSION: The carbon nanotubes in chitosan/collagen-based composite can be an ideal candidate for peripheral nerve regeneration.

Objective To explore the clinical features and treatment of thyroid carcinoma in children. Method The clinical data of 19 children under 14 years old with thyroid carcinoma diagnosed and treated from January 2003 to January 2014 were retrospectively analyzed. Results In 19 cases (12 males and 7 females), there were 18 cases of papillocarcinoma and one case pf follicular carcinoma. Unilateral lobectomy plus isthmectomy was performed in 6 cases, subtotal thyroidectomy in 4 cases and total thyroidectomy in 9 cases. Unilateral cervical lymph node dissection was performed in 5 cases and bilateral in 11 cases. After the operation, multiple lesions were confirmed by pathology in 9 cases, thyroid capsular invasion in 14 cases, lymphatic metastasis in 15 cases and distant metastasis in 5 cases. All the patients were treated with TSH, and 10 cases were treated with 131I after operation. The median follow-up time was 63 months. There was no death in all cases, while local residual tumor recurrence was found in 2 cases and cervical lymph node metastasis in 2 cases and distant metastasis in one case. Conclusion Thyroid carcinoma in children is mostly well-differentiated, so the overall prognosis is better. However, children who have extracapsular invasion, multiple lesions in bilateral thyroid, cervical lymph node metastasis and distant metastasis are at high risks and should be treated with comprehensive therapy that includes total thyroidectomy.

Objective To investigate the influence of 18F-FDG on the proliferation of Lewis lung cancer cell line,and to elucidate its possible mechanism.Methods Morphological changes of cells after culture for 24 h at different concentrations of 0,0.37,1.85,3.70 and 7.4 (×106) Bq/ml of 18F-FDG were observed by using inverted microscopy and electron microscopy.The apoptosis and phase distribution of cell cycle of irradiated cells were analyzed with flow cytometry.DNA synthesis of irradiated cells was assayed by 3H-TdR incorporation.Lipid peroxidation was measured by chromometry and expression of Bcl-2 and Bax protein was measured by immunohistochemical technique.Results Exposed to (0-7.40) × 106Bq/ml of 18F-FDG for 24 h,the cumulative absorbed doses delivered to cells in five groups were 0,0.11,0.55,1.10 and 2.20 Gy,respectively.Irradiated cells showed morphological changes of apoptosis.The apoptosis rate of irradiated cells was increased from (4.05 ± 0.01)% to (25.6 ± 0.28) % (t = 188,P＜0.01).3H-TdR incorporation rate was decreased from 100% to(22.0 ± 0.51)% (t =27.6,P ＜0.05).The levels of M DA in cells were augmented from (0.08 ± 0.03) to (0.67 ± 0.12) μmol/L (t =11.7,P ＜ 0.01).Cell cycle arrest was found in G2/M phase with the increasing doses from 0 to 2.20 Gy.The expression of Bcl-2 protein was decreased while that of Bax protein increased.Conclusions 18F-FDG could induce the apoptosis of cells and inhibit the proliferation of cells.

Objective To construct a mouse model for real-time,noninvasive and specific monitoring of inflammation activation in hepatic tissues.Methods An inflammation reporter gene was targeted to the liver by hydrodynamic gene delivery technology.Bioluminescence imaging was used to detect the firefly luciferase(Fluc) expression in the mouse liver after inflammatory stimulation.Besides,the relevance between the light intensity and inflammation level was also intensively investigated.Results pIL-6-Fluc was successfully delivered to the liver.The hydrodynamic gene delivery could cause a transient liver injury that could return normal in 5 to 7 days.The expression of pIL-6-Fluc could be induced by lipopolysaccharides(LPS) treatment with an about (46.80±13.35) fold increase at the peak value,which was significantly higher than that detected by ELISA [(4.09±0.96)fold].Conclusion An inflammation reporter mouse model is constructed in this study by hydrodynamic gene transfection,allowing noninvasive monitoring of inflammation activation specifically in hepatic tissues.The reporter model is capable of monitoring inflammation activation with a sensitivity higher than that of ELISA.

OBJECTIVETo compare the cytocompatibility of two kinds porous bioactive glass-ceramic made by same raw materials.

METHODSApatite/wollastonite bioactive glass-ceramic (4006) were prepared by sol-gel method, and bioactive glass (45S5) were prepared by melting method. Bone marrow stromal cells (BMSCs) were cultivated, differentiated and proliferated into osteoblasts, from a rabbit's marrow in the differentiatiofn culture medium with active function. The viability of BMSCs cultivated with extraction of these two kinds of biomaterial, which could represent the cytotoxicity effect of 4006 and 45S5 against BMSCs, was evaluated by the MTp assay. BMSCs were seeded and cocultivated with two kinds of biomaterial scaffolds respectively in vitro. The proliferation and biological properties of cells adhered to scaffolds were observed by inverted phase contrast microscope, scanning electron microscope (SEM), and environmental scanning electron microscope (ESEM), and a suitable cell amount for seeding on the scaffold was searched.

RESULTSThere was no difference on the viability of BMSCs only cultured for one day by complete extract of 4006 and culture medium (P>0.05), but there was significant difference between them when the cells had been cultured for 3 days(P<0.01). The extract of 45S5 had significantly higher cytotoxicity than extract of culture medium (P<0.01). The BMSCs adhered, spread, and proliferated throughout the pores of the scaffold 4006, and the amount of cells adhered to 4006 was more than to 45S5. The adhered cells to 4006 increased with the rising amount of cells seeded. And 2 x 10(7) cells.mL-1 suspension resulted inthe highest cell adherence during the comparative cells adherence test.

CONCLUSIONApatite/woolastonite bioac tive glass-ceramic has good bioactivity and cytocompatibility. Therefore, it may have the potential to be a new cell vehicle for bone tissue engineering. And the suitable seeding cell amount of apatite/wollastonite bioactive glass-ceramic should be 2x10(7) cells.mL-1 or even more than that.

Animals ; Biocompatible Materials ; Calcium Compounds ; Cell Adhesion ; Cell Differentiation ; Ceramics ; Glass ; In Vitro Techniques ; Mesenchymal Stromal Cells ; Osteoblasts ; Rabbits ; Silicates ; Tissue Engineering

Objective:To investigate the factors related to recurrence and prognosis of retroperitoneal liposarcoma.Method:The clinical data of patients with primary retroperitoneal liposarcoma who underwent surgical treatment in the First Affiliated Hospital of Zhengzhou University from June 2011 to January 2020 were analyzed retrospectively. There were 42 males and 47 females and patients’median age was 53 (26-78). Sixty-five cases were treated by operation in our hospital, and 24 cases were primarily treated by the operation in another hospital. The clinical manifestations of the initial diagnosis included retroperitoneal mass in 41 cases, abdominal distension in 12 cases, abdominal pain in 10 cases, fever in 11 cases, nausea, vomiting and poor appetite in 8 cases, frequent urination and dysuria in 6 cases, and bilateral lower limb edema in 1 case. Preoperative CT imaging showed that the tumor body was located in the retroperitoneal kidney area in 58 cases, while in the retroperitoneal space or the pelvic extraperitoneal space in 31 cases. There were 55 single cases and 34 multiple cases. The median tumor length was 20(3-52) cm. Among the primarily treated 65 patients, 47(72.3%) were considered as primary retroperitoneal liposarcoma by preoperative imaging examination. Among the 89 patients treated by surgery, 78 underwent endoscopic surgery, among which 21 underwent laparoscopic surgery, 38 cases of retroperitoneal laparoscopic surgery, 19 cases of Da Vinci robot-assisted laparoscopic surgery. Open operation was performed in 11 cases. There were 87 patients undergoing radical resection and 2 patients undergoing palliative resection. Forty-two patients underwent intraoperative combined resection of the adjacent organs. The recurrence and survival status of patients were followed up.Results:All the 89 patients underwent the operation successfully, with the median operative blood loss of 200 (10-2000) ml. There were 23 cases being diagnosed of well differentiated liposarcoma, 40 cases of dedifferentiated, 20 cases of myxoid/round, 5 cases of myxoid liposarcoma, and 1 cases of mixed type. Pathologically, there 42 cases with low grade histology and 47 cases with high grade histology. In this study, 89 patients were followed up for 3 to 108 months, and the median follow-up time was 28 months. The 5-year recurrence free survival rate, disease-free survival rate and overall survival rate of the patients were 16.7%, 16.1% and 52.6%, respectively. There were 57 patients presenting local recurrence, 1 patient of lung metastasis, and 1 patient of liver metastasis, and the median disease-free survival time was 24 months. There were 42 patients died of the disease, with a median survival time of 64 months. Univariate analysis showed that intraoperative blood loss( P<0.01), whether multiple cases( P<0.01), pathologic types( P<0.01), and histological grades ( P<0.01) were related to disease-free survival.The intraoperative blood loss( P<0.01), multiple cases( P<0.05), pathologic types ( P<0.05), and recurrence ( P<0.01)were related with overall survival. Gender, age, tumor size, tumor location, whether primary surgery, radical resection or combined resection of adjacent organ had no effect on the prognosis of patients ( P>0.05). Cox regression model multivariate analysis revealed that surgical bleeding ( RR=2.360, 95% CI 1.313-4.241, P=0.004), multiple tumor ( RR=1.899, 95% CI 1.068-3.375, P=0.029), and pathological type ( RR=4.976, 95% CI 1.622-15.264, P=0.005) were independent factors affecting disease-free survival. The recurrence was an independent factor affecting the overall survival of patients ( RR=31.495, 95% CI 1.062-933.684, P=0.046). Conclusions:Retroperitoneal liposarcoma is a rare disease with high recurrence rate. The intraoperative blood loss, whether multiplicity and pathological type are independent factors affecting the disease-free survival, and recurrence is independent factors affecting the overall survival.

OBJECTIVETo determine the optimized condition under which BMP expression vector will be constructed to transfect bone marrow stromal cells (MSCs), plasmid vector coding enhanced green fluorescence protein (EGFP) gene pEGFP was transferred into MSCs. The transfer efficiency and transient expression were subsequently tested.

METHODSpEGFP plasmid was amplified and tested by an enzyme cutting technique in vitro. MSCs, which were initially obtained from the bone marrow of rabbits, were cultured in vitro and transferred with pEGFP by means of lipofectamine media methods. The ratio of plasmid and lipofectamine was varied according to the experiment design. Transfer efficiency and transient expression were evaluated by fluorescent microscopy.

RESULTSTransfer efficiency was correlated with the ratio of plasmid and lipofectamine. The expression of EGFP began in 24 hours after transferring, reached maximum in 48-72 hours and decreased in 1 week, however there remained a weak expression for more than 3 weeks.

CONCLUSIONThe efficiency of transferring pEGFP into MSCs could achieve to 30% with proper ratio of plasmid and lipofactamine. pEGFP was an ideal transient expression vector for MSCs gene transference.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Bone Morphogenetic Proteins ; genetics ; Cells, Cultured ; Genetic Vectors ; Green Fluorescent Proteins ; Lipids ; genetics ; Luminescent Proteins ; biosynthesis ; genetics ; Plasmids ; genetics ; Rabbits ; Stromal Cells ; cytology ; metabolism ; Transfection