Objective:To construct a recombinant adenoviral vector carrying the specific siRNA of Survivin gene,and to observe its effect on the expression of Survivin gene and on the radiosensitivity of cancer cells in tumor-bearing mice.Methods:The specific siRNA of Survivin gene was designed and synthesized,and a recombinant adenovirus AdEGFP-siRNA was subsequently constructed.SMMC 7721 xenograft models were established with nude mice and were divided into the following 5 groups:siRNA+radiotherapy and siRNA groups(intratumoral injection of AdEGFP-siRNA),siRNA(-)group(injected every other day with AdEGFP-siRNA[-],2?108 pfu/100 ?l per time,total 5 times),pure radiotherapy group and blank control groups(injected with the same volume of normal saline).On day 10,12,14,and 16,the mice in siRNA+radiotherapy and pure radiotherapy groups were given 5 Gy/time radiotherapy.The tumor volumes were measured regularly.The expression of Survivin in tumor tissues was determined immunohistochemically.Results:Adenovirus AdEGFP-siRNA harboring the specific siRNA of Survivin gene and enhanced green fluorescent protein gene(EGFP)was successfully recombined.The growth of SMMC 7721 xenografts in nude mice was inhibited after injecting AdEGFP-siRNA,with the inhibition rate being 56.2%.The inhibition rate in AdEGFP-siRNA therapy + radiotherapy increased to 82.6%.Immunohistochemistry study showed that the specific siRNA markedly silenced the expression of Survivin gene in hepatocarcinoma cells.Conclusion:The specific siRNA can markedly silence Survivin gene and subsequently inhibit the growth of cancer;meanwhile,it can also increase the radiosensitivity of cancer cells so as to improve the treatment effect.

OBJECTIVE:To prepare core-shell nanocapsules loading nicardipine hydrochloride and to investigate its pharmaceutical characteristics. METHODS: Core-shell nanocapsules were prepared using layer-by-layer electrostatic self-assembly technique. The indexes including the shape and particle size and the loaded drug amount of the nanocapsules were evaluated, and its accumulative release rates in artificial gastric juice and intestinal juice were computed and compared with those of its crude drug. RESULTS: The results showed that the nanocapsules were spherical with a mean particle size of 200 nm and a maximum loaded drug amount of 2.512%. The drug release rate within 12 h reached 18.64% in artificial gastric juice and 70% in artificial intestinal juice, whereas within 3 h the drug release rate of its crude drug in artificial intestinal juice reached 87%. CONCLUSIONS: The prepared core-shell nanocapsules containing nicardipine hydrochloride had a good pharmaceutical property.

Purpose To explore the CT features of extra-adrenal ganglioneuroma (GN), and to improve the diagnostic accuracy of the disease. Materials and Methods CT features of 13 patients with extra-adrenal GN confirmed by surgical pathology were retrospectively analyzed, and compared with pathological findings. Results Two of 13 lesions were located in neck, four in posterior mediastinum, four in retroperitoneal space, two in front of sacrum, one in the region of renal hilus. All these masses were well defined, two appeared as round, three as oval, 8 as irregular shape. Of eight cases with irregular shape, six lesions presented a tendency of wedging into the space encasing and lapsing the vessel, with no evidence of invasion of the surrounding organs or vessels. Two tumors grew along the vertebrae and invaded intervertebral foramen. On plain CT imagings, the attenuations of the tumors were less than that of muscle, the masses with homogeneous and nonhomogeneous density were found in four and nine cases, respectively. Two patients associated with scattered and patchy cafilication. After contrast administration, eight lesions showed inhomogeneous enhancement:slight enhancement were seen in the arterial phase and gradually more strong enhancement during the portal venous and delayed phase. Enhancing lines or patchy were observed inside these tumors. Pathologically, the tumors appeared on a large amount mucus background, a few of scattering or nesting ganglion cells were distributed in many mature spindle cells, which lined in weave or fasciculation. Conclusion CT manifestations of extra-adrenal GN have certain characteristics, which are correlated closely with pathological features, therefore is helpful for the diagnosis and differential diagnosis.

The Interleukin-2 gene cDNA was cloned into the Pichia pastoris expression vector pPICZB,which is under the control of the alcohol oxidase promoter AOX1. The linearized recombinant plasmid of BoIL2-pPICZB,digested by Sac I ,was transformed into X-33 strains by electroporation. The multi-copy insert transformants were screened by Zeocin-resistance and induced by 1% methanol. The intracellular expression products were tested by SDS-PAGE analysis and Western blotting. Purified recombinant BoIL2 was gained by metal-chelating affinity chromatographic (MCAC). Assay with murine CTLL-2 cells showed that the recombinant BoIL2 exhibited the biological activity.

Objective To observe the protective effect of ghrelin on hippocampal injury induced by global cerebral ischemia/reperfusion (I/R) and explore its effect mechanisms.Methods The male Sprague-Dawley (SD) rats were randomly divided into four groups,namely sham group,I/R group,normal saline (NS)+I/R group and Ghrelin+I/R group,with 42 rats in each group.The model of I/R was reproduced by clipping bilateral carotid artery of rats 15 minutes and then releasing them for 60 minutes.There were no challenges for rats in sham group,just exposed their carotid artery.Ghrelin+I/R group and NS+I/R group were challenged by injecting 1 μ.L ghrelin or NS into lateral ventricle before I/R.Some of brain tissue in the rats was harvested after experiment to determine the levels of malonaldehyele (MDA),myeloperoxidase (MPO) and glutathione (GSH) in hippocampus by using chemical colorimetry and observe infarct sizes and histopathology.Single extracellular neuron discharge in other rats was recorded to observe the activity of glutamic sensitive neurons (Glu-N) and γ-aminobutyric acid (GABA) sensitive neurons (GABA-N) in hippocampus CA1 region of rats suffered I/R.Results Compared with sham group,the levels of MDA and MPO in hippocampus of rats in the I/R group were raised markedly,the level of GSH was decreased significantly,the infarct sizes was increased significantly and pycnosis neurons were increased markedly.All sorts of indexes between NS+I/R group and I/R group showed no significantly statistical significance.Compared with NS+I/R group,the levels of MDA and MPO in hippocampus of rats in the Ghrelin+I/R group were decreased significantly [MDA (nmol/g):16.4 ± 4.2 vs.24.5 ± 6.7,MPO (nmol/g):6.4 ± 1.8 vs.10.2 ± 2.9,both P ＜ 0.05],the activity of GSH was risen remarkably (μmol/g:2.65 ± 0.72 vs.1.66 ± 0.50,P ＜ 0.05),the infarct sizes of hippocampus were reduced markedly [(43.9 ± 9.5)％ vs.(77.0 ± 12.7)％,P ＜ 0.01],the number of pycnosis neuron was reduced markedly (cells:36.2±4.5 vs.47.1 ±6.1,P ＜ 0.01).The results of electrophysiology showed that the discharge frequency of Glu-N and GABA-N in hippocampus CA1 region of rats in I/R group increased markedly as compared with sham group,and no significant difference in the discharge frequency of Glu-N and GABA-N between NS+I/R group and I/R group.Compared with NS+I/R group,injected ghrelin could make the discharge frequency of Glu-N in hippocampus CA1 region of rats decreased markedly (Hz:3.81 ±0.67 vs.4.98±0.33 at ischemia,3.01 ±0.37 vs.3.77 ± 0.41 at reperfusion,both P ＜ 0.05),and the discharge frequency of GABA-N increased markedly (Hz:5.62 ± 0.54 vs.3.62±0.39 at ischemia,4.81±0.48 vs.3.71±0.21 at reperfusion,both P ＜ 0.05).Conclusion Ghrelin might protect hippocampal neuron after I/R iniury,and neuron excitability decrease might be related.

Objective To research the functional role of thyroidal motilin and the effects of electric excitation of the paraventricular nuclei(PVN) on gastric motility and the levels of motilin in thyroid and plasma.Methods The expression of motilin in rat and human thyroid was detected by immunofluorescence staining.A phase Ⅲ-like contraction was recorded before and after thyroidectomy and after PVN excitation.The changes in concentrations of plasma FT3,FT4 and motilin were determined via radioimmunoassay (RIA).c-Fos expression of PVN after thyroidectomy and motilin expression in thyroid after PVN excitation were observed by immunohistochemical staining.Results There were motilin immunoreactive cells in rat and human thyroid.The phase Ⅲ-like contraction and concentration of motilin in plasma decreased significantly when measured on the second and fourth days after thyroidectomy(2d,P＜0.01 ;4d,P＜0.05).The expression of c-Fos in PVN after thyroidectomy was significantly increased(P＜0.05).An electric excitation of PVN could increase the concentration of motilin in plasma and thyroid and increase corresponding gastric motility in rats (P ＜0.05).The increased phase Ⅲ-like contraction by PVN excitation could be partially inhibited by administration of motilin receptor antagonist,GM-109 (P＜0.05).Excitation of PVN in thyroidectomized rats resulted in lower plasma motilin and less intense phase Ⅲ-like contraction of stomach,as compared with the sham operated control group(P＜0.05).Conclusion Motilin from the thyroid may be secreted into the peripheral plasma to affect gastric motility and PVN may modulate gastric motility and motilin expression in the thyroid.

Objective To investigate the tracking neurofibra pathway from the hippocamlpal neurons to septal nucleus,and to explore the effects and mechanisms of ghrelin on the learning and memory in septal nucleus lesion rats.Methods Retrogradely tracing method was used to observe Fluorogold (FG) reaching sites in hippocampus.The septal nucleus was destructed by direct current using stereotactic technique.Step-down test and morris water maze were used to test the effects of learning and memory ability by means of microinjecting ghrelin into hippocampal CA1 area in rats.Results After injection of FG into septal nucleus,retrogradely labeled neurons and neurofibra were found in the hippocampal neurons with FG.Ghrelin injection of hippocampal CAI area could promoter learning and memory ability in rats.It showed that the escaped latent period was significantly lengthened ( (3.2 ± 0.9) s vs (6.9 ± 1.1 ) s,P ＜ 0.05) and the wrong numbers in 5 min were obviously decreased in escape response test; and the latency of looking for the plat was significantly shorter in morris water maze test ( 1.8 ±0.4vs 0.8 ± 0.1,P ＜ 0.05 ).However the effects above-mentioned on learning and memory was significantly weak after septal nucleus lesioning.It showed that the escaped latent period was markedly shortened ( ( 19.5 ±3.2)s vs ( 10.5 ± 2.1 ) s,P ＜ 0.05 ) and the wrong numbers in 5 min were obviously increased ( ( 3.9 ± 0.8 ) s vs ( 1.8 ±0.5 ) s,P＜0.05 ) in escape response test.The latency of looking for the plat was significantly lengthened in morris water maze (P＜0.05).Conclusion Ghrelin could elevate the learning and memory ability in hippocampus,and the effects may be related to the septal-hippecampus pathway.

Adenomyomatosis of the gallbladder (GAM) is an acquired,benign proliferative lesion of the gallbladder which is characterized by mucosal proliferation with invaginations and diverticula penetrating into the thickened muscular layer (Rokitansky-Aschoff sinuses).GAM consists of 3 types:diffuse,segmental and fundus GAM.There is no specific presentation of GAM,and computed tomography is helpful for the diagnosis of this disease.From July 2010 to May 2013,16 patients with fundus GAM were admitted to the Second People's Hospital of Wuxi.Rokitansky-Aschoff sinuses and calotte sign at the thickened muscular layer of the fundus of the gallbladder are the typical presentation of the fundus GAM.Enhanced computed tomography examination is of great importance for the diagnosis of the fundus GAM.

Objective:This study aimed to explore the effects ofnesfatin-1 on gastric distension (GD)-sensitive neurons in the basomedial amygdala (BMA) and the potential mechanism for nesfatin-1 to regulate gastric motility through the arcuate nucleus (Arc).Methods:The projection of nerve fiber and expression of nesfatin-1 were observed by retrograde tracing and fluo-immunohistochemistry staining;The nuclei microinjection and nuclei electrical stimulation,extracellular discharges of single unit neuron were used to observe the effects ofnesfatin-1 on the GD neurons;Gastric motility recording in vivo were used to monitor the effects ofnesfatin-1 on the amplitude of constriction and frequency of gastric motility in conscious rats.Results:NUCB2/Nesfatin-1/fluorogold-double labeled neurons were from ARC to BMA;Nesfatin-1 could excited the firing rate of most of the GD-E neurons (4.25± 1.02 Hz vs.5.32± 1.17 Hz,P＜0.01) and decreased the firing rate of most of the GD-I neurons (3.73± 0.92 Hz vs.2.64± 0.86 Hz,P＜0.01),inhibited the gastric motility,amplitude and frequency,SHU9119 could weaken the responses induced by nesfaton-1;Electrical stimulation of the Arc,the firing rate of nesfatin-1-induced GD-response neurons (GD-E:5.14± 1.32 Hz vs.6.75± 1.84 Hz,P＜0.05;GD-I:2.84± 0.86 Hz vs.4.05± 1.12 Hz,P ＜0.05) and the gastric amplitude and frequency were increase.Conclusion:It was suggested that nesfatin-1 in the BMA plays an important role in decreasing gastric motility and the Arc may be involved in this regulation process.

Objective:To investigate the effects of Orexin peptides on feeding and energy metabolism in mice.Methods:The mice were divided into two groups:feeding group and metabolic group.The feeding group were injected with different doses (1,3 and 10 nmol) of orexin-A and orexin-B to observe their effects on feeding and the activity of tyrosine hydroxylase in liver.We used the metabolic cages and observed the changes of respiration rate and respiration rate of mice were under light condition,dark condition and fasting condition.Results:Compared with the control group,1 nmol and 10 nmol orexin-A significantly stimulated mice to feed (P ＜0.05) within 4 hours after injection,and the effect of 3 nmol orexin-A on feeding was not obvious,but increase the activity of tyrosine hydroxylase.Any dose of orexin-B did not show a stimulating effect on mice feeding.(P ＞0.05).In the light cycle,orexin-A could significantly reduce the respiration rate (RQ),the metabolic rate was significantly increased (P ＜0.05);In the dark cycle,orexin-A had no effect on RQ,but the metabolic rate was significantly rised (P ＜0.05);But the injection of orexin-A in fasting mice induced a brief increase in RQ and a significant increase in metabolic rate (P ＜0.05).Conclusion:Orexins may play an important role in regulating feeding and energy metabolism in mice.