Objective To investigate the effect of over-expression of human ribonuclease inhibitor suppresses invasion and migration of transplanted bladder cancer .Methods The T24 cells were stably transfected with pIRES2-EGFP-RI and pIRES2-EG-FP plasmid respectively .Using the cell transfected with pIRES2-EGFP and untransfected cell as controls .and the positive clones were screened by G418 ,respectively ;Tumor cells of the three groups at 2 × 106 were respectively injected into the back of BALB/C nude mice to establish the xenograft models .Change of micro-blood vessels in tumor tissue and expression of CD31 were detected by Immunohisto-chemical and HE staining .Immunohisto-chemical assay was used to detect the expression of RI ,MMP-2 ,MMP-9 ,E-cadherin ,N-cadherin ,Vimentin ,Snail ,Slug ,Twist in the tumors .Results Animal experiment showed that the T24-RI cells group significantly inhibited the growth of bladder cancer compared with the other two control groups .Compared with the T24 and T24 vector cells groups ,the microvessel density in tumor tissue of T24-RI group was notably reduced and the expressions of MMP-2 , MMP-9 ,N-cadherin ,Vimentin ,Snail ,Slug ,Twist significantly were decreased simultaneously ,while the expressions of RI and E-cadherin were increased .Conclusion up-regulation RI can inhibit the growth of transplanted bladder cancer in nude mice by decrea-sing the expression of invasion protein and EM T protein .

Objective To describe the structure and the level of the stress of nursing students in clinic practice,and supply reference for reform of clinical teaching method.Methods Totally 100 undergraduate nursing students in clinic practice,who would graduate from nursing school and begin to work in 2013,were interviewed and investigated.Results The main stress sources were divided into five aspects:recognition,clinical nursing ability,career development,field response and exhaustion of body and mind.Conclusions The structure and level of the stress of nursing students changed a lot.Some new appropriate reform should be introduced into clinic education,in order to help nursing students to complete psychological change and role adaptation.

OBJECTIVETo observe the phenomenon of delayed neuron death secondary to traumatic brain injury, and the protective effect of p53 antisense oligonucleotide against neuron apoptosis secondary to traumatic brain injury.

METHODSThis study was based on the rat diffuse brain trauma model established by a simple weight-drop device. The behavior scales of neural function of rats and TUNEL that examined the injury extent of DNA in the cortex were used as a general assessment of brain injury. Electron microscope was used to observe the histological and cellular morphology. mRNA and protein expression of p53 were detected by in situ hybridization and immunohistochemistry. In this model, the therapeutic effect of p53 antisense drug were observed.

RESULTSFollowing trauma, the behavior scores of rats decreased rapidly and remarkably. Apoptotic neurons appeared in the traumatized cortex as early as 2 hours after impact, and peaked at 24 hours. As early as 2 hours after impact, p53 mRNA and p53 protein in the cortex were expressed increasingly.

CONCLUSIONNeural apoptosis plays an important role in delayed neuron death and is responsible for neural dysfunction following traumatic brain injury. So the inhibition of neural apoptisis would turn into a new therapic measure against delayed neuron death following traumatic brain injury.

Animals ; Apoptosis ; drug effects ; Brain Injuries ; physiopathology ; Female ; Immunohistochemistry ; In Situ Hybridization ; In Situ Nick-End Labeling ; Male ; Microscopy, Electron ; Neurons ; drug effects ; pathology ; ultrastructure ; Oligonucleotides, Antisense ; pharmacology ; Rats ; Rats, Wistar ; Tumor Suppressor Protein p53 ; genetics ; metabolism

From ancient times to the present,acupuncture has made tremendous contributions to women' s reproductive health.Based on traditional Chinese medicine of kidney-menstruation-Chong and Ren-uterus axis and Western medicine of hypothalamus-pituitary-ovarian axis comparative analysis study,try to explore possible ways from the acupuncture point to molecular changes in the process of acupuncture and moxibustion treatment.

Objective To investigate the effects and mechanisms of 1,25-Dihydroxyvitamin D3 [1,25(OH)2 D3] on hyperoxia-induced lung injury of neonatal rats.Methods Neonatal Sprague-Dawley rats were randomly divided into air group,hyperoxia group and 1,25(OH)2D3 group within 12 hours after birth,eight in each group.Rats in air group were exposed to air,while those in hyperoxia and 1,25 (OH) 2 D3 group were exposed to hyperoxia (≥85 ％ oxygen concentration).Rats in 1,25(OH)2D3 group were injected with 1,25(OH)2D3 0.5 μg/(kg · d) intraperitoneally once a dayfor seven days,meanwhile the rats in the other two groups received 0.9 ％ saline in the same way.All rats were sacrificed on day 7.Lung tissue sections were HE stained in order to assess lung histological changes and lung radical alveolar count (RAC).The levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β and IL-6 mRNA were measured by real time-polymerase chain reaction.Analysis of variance and LSD test were applied for statistics.Results Compared with the air group,the weight of rats in the hyperoxia group was significantly lower on day 7 [(8.48±1.34) g vs (12.51±0.47) g,t=8.05,P＜0.05],while the weight of rats in 1,25(OH)2D3 group [(10.29±1.00) g] was higher than that in the hyperoxia group (t=3.61,P＜0.05).Lung tissue structure was normal in the air group.In the hyperoxia group,inflammatory exudation was observed in pulmonary interstitial,the alveolar size was uneven,and the RAC was lower than that in the air group (5.6±0.1 vs 6.8±0.2,t=21.45,P＜0.05).The RAC in 1,25(OH)2D3 group (6.2±0.1) was significantly increased compared with that in the hyperoxia group (t=11.76,P＜0.05),but still lower than that in the air group (t=9.69,P＜0.05).The expressions of TNF-α,IL-1β and IL-6 mRNA in hyperoxia group (0.0348±0.0006,0.0269±0.0003 and 0.0368 ± 0.0006) were higher than those in the air group (0.0111±0.0007,0.0040±0.0003 and 0.0162 ±0.0007,t=56.54,111.12 and 49.26,P＜0.05,respectively).The expressions of TNF-α,IL-1β and IL-6 mRNA in the 1,25 (OH)2D3 group (0.0203±0.0009,0.0141±0.0004 and 0.0251±0.0009) were lower than those in the hyperoxia group (t=34.44,61.93 and 27.99,P＜0.05,respectively),but higher than those in the air group (t=22.10,49.19 and 21.27,P＜0.05,respectively).Conclusions 1,25(OH)2D3 could attenuate hyperoxia-induced lung injury by inhibiting the expression of inflammatory cytokines.

Purpose To study the malignant transformation after treating rat oval cell line ( WB-F344 ) with chemical carcinogen N-methyl-N′-nitro-N-nitrosoguanidine ( MNNG) . Methods WB-F344 cells were cultured with MNNG for severe times. The biological characteristics of induced cells were detected through the following methods:to check proliferation activity by flow cytometry analysis, to examine malignant transformation degree of induced cells by soft agar assay and tumor formation in NOD/SCID mice, and to investi-gate the transcriptional and protein levels of hepatocellular carcinoma marker GGT, GST-P by real time-PCR. Results Oval cells in-duced by MNNG showed changes in biological characteristics and malignant molecular markers. Conclusion Hepatic oval cells model is successfully established, which can be confirmed by tumor formation in NOD/SCID mice.

Objective To explore the changes of Sema3A and it′s receptor Npl in temporal lobe epilepsy(TLE)rat brain and the roles in epileptogenesis mechanism.Methods TLE model was established with male healthy SD rats,in which mossy fiber sprouting(MFS)was verified using Neo-Timm staining method.Sema3A mRNA,Npl mRNA and protein was respectively analyzed by immunohistochemistry and in situ hybridization in the entorhinal cortex(EC)or dentate gyrus(DG)at different time after LiCL-PILO induced TLE.Results There were Mossy fiber sprouting(7d:0.70?0.42,15d:1.50?0.52,30 d:2.20 ?0.41,60 d:2.50?0.51)in DG inner molecular layer(IML)of TLE rat compared with those of controls (P

1H NMR metabonomics approach was used to reveal the chemical difference of urine between patients with Xiao-Chaihu Tang syndrome (XCHTS) and healthy participants (HP). The partial least square method was used to establish a model to distinguish the patients with Xiao-Chaihu-Tang syndrome from the healthy controls. Thirty-four endogenous metabolites were identified in the 1H NMR spectrum, and orthogonal partial least squares discriminant analysis showed the urine of patients with Xiao-Chaihu Tang syndrome and healthy participants could be separated clearly. It is indicated that the metabolic profiling of patients with Xiao-Chaihu Tang syndrome was changed obviously. Fifteen metabolites were found by S-pot of OPLS-DA and VIP value. The contents of leucine, formic acid, glycine, hippuric acid and uracil increased in the urine of patients, while threonine, 2-hydroxyisobutyrate, acetamide, 2-oxoglutarate, citric acid, dimethylamine, malonic acid, betaine, trimethylamine oxide, phenylacetyl glycine, and uridine decreased. These metabolites involve the intestinal microbial balance, energy metabolism and amino acid metabolism pathways, which is related with the major symptom of Xiao-Chaihu Tang syndrome. The patients with Xiao-Chaihu Tang syndrome could be identified and predicted correctly using the established partial least squares model. This study could be served as the basis for the accurate diagnostic and reasonable administration of Xiao-Chaihu-Tang syndrome.
Humans ; Least-Squares Analysis ; Medicine, Chinese Traditional ; Metabolome ; Metabolomics ; Proton Magnetic Resonance Spectroscopy ; Syndrome ; Urinalysis

OBJECTIVETo observe human neuronal apoptosis secondary to traumatic brain injury, and to elucidate its regulative mechanism and the change of expression of apoptosis-related genes.

METHODSSpecimens of brain were collected from cases of traumatic brain injury in humans. The histological and cellular morphology was examined by light and electron microscopy. The extent of DNA injury to cortical neurons was detected by using TUNEL. By in situ hybridisation and immunohistochemistry the mRNA changes and protein expression of Bcl-2, Bax, p53, and caspase 3 p20 subunit were observed.

RESULTSApoptotic neurons appeared following traumatic brain injury, peaked at 24 hours and lasted for 7 days. In normal brain tissue activated caspase 3 was rare, but a short time after trauma it became activated. The activity peaked at 20-28 hours and remained higher than normal for 5-7 days. There was no expression of Bcl-2 mRNA and Bcl-2 protein in normal brain tissue but 8 hours after injury their expression became evident and then increased, peaked at 2-3 days and remained higher than normal for 5-7 days. The primary expression of Bax-mRNA and Bax protein was high in normal brain tissue. At 20-28 hours they increased and remained high for 2-3 days; on the 7th days they returned to a normal level. In normal brain tissue, p53mRNA and P53 were minimally expressed. Increased expression was detected at the 8th hour, and decreased at 20-28 hours but still remained higher than normal on the 5th day.

CONCLUSIONSFollowing traumatic injury to the human brain, apoptotic neurons appear around the focus of trauma. The mRNA and protein expression of Bcl-2, Bax and p53 and the activity of caspase 3 enzyme are increased.

Adult ; Brain Injuries ; pathology ; physiopathology ; Caspase 7 ; Caspases ; metabolism ; Female ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Male ; Necrosis ; Neurons ; pathology ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein

OBJECTIVETo investigate the specificity of the acupoints and related factors on acupuncture effects.

METHODSThe rat model of cerebral median artery occlusion (MCAO) was established with thread ligation according to Zea-Longa method. The acupoint group and the non-acupoint group were stimulated by acupuncture with 9 different parameters (marked group No. I - VI) combined by frequency (60 Hz, 120 Hz, 180 Hz) and time (5 s, 60 s, 180 s). The acupoint of "shuigou" (GV 26) was selected for the acupoint group, the fixed point which was at a distance of 10 mm from the iliac crest below the ribs in the affected side. The acupuncture effect was evaluated with the percentage of brain infarct area after TTC dyeing.

RESULTS(1) Acupoint groups could obviously reduce the brain infarction area in MCAO rats. Under 9 intervention parameters, acupoint groups had much more effectiveness tendency than that of the non-acupoint groups. (2) Acupoint group VI was the most effective in reducing the brain infarction area among 9 acupuncture parameter groups.

CONCLUSION(1) Acupoint has specificity effect on reduction of brain infarct area in MCAO rats. (2) Acupoint group VI (180 Hz; 5 s) could significantly reduce the percentage of cerebral infarction area by TTC dyeing.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Brain Infarction ; therapy ; Disease Models, Animal ; Humans ; Male ; Random Allocation ; Rats ; Rats, Wistar