Objective To study the effect of the inhibitor of apoptosis protein,Livin on proliferation and multi?drug resistance of lung adenocarcino?ma cells A549. Methods A549 cells were transfected with the eukaryotic expression vector pcDNA3.1?Livin. A549 cell clone with stable expres?sion of Livin was obtained through G418 screening. Expressions of Livin mRNA and protein in the transfected cells were respectively measured by re?verse transcription polymerase chain reaction(RT?PCR)and Western blot. The distribution of cell cycle phase was determined using flow cytometry. The level of P?gp mRNA and protein in A549 cells transfected with pcDNA3.1?Livin was detected by RT?PCR and Western blot. The analysis of multi?drug resistance of A549 treated with different chemotherapeutics was performed by MTT. Results The mRNA and protein expressions of Liv?in were both significantly increased in the transfected A549 cells. The flow cytometry analysis showed there was higher percentage of S phase and low?er percentage of G0/G1 phase in A549 cells transfected with pcDNA3.1?Livin. Compared with control groups,the expression of P?gp mRNA and pro?tein was increased in A549 cells transfected with pcDNA3.1?Livin,which showed a higher drug resistance and lower sensitivity to chemotherapic drugs such as ADM,MTX,CTX,and DDP(P<0.05). Conclusion Overexpression of Livin could enhance the proliferation of A549 cells,and high expression of P?gp caused by Livin could serve as one of the causes for multi?drug resistance in lung adenocarcinoma against chemotherapies.

Objective To investigate the effects of small interfering RNA (siRNA) silencing Nanog gene on the ability of migration and invasion of the human lung adenocarcinoma A549 cells. Methods The human lung adenocarcinoma A549 cells were transfected with siRNA targeting Nanog gene , and three experiment groups were set up. The expression level of Nanog was detected using reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. Cell migration was examined by wound healing assay and cell invasion was detected by Transwell assay. Results The Nanog silencing cell group (A549-siNanog) showed much lower level of Nanog mRNA and protein (0.40 ± 0.06, 0.50 ± 0.03) than A549-siNC cell group (0.97 ± 0.03, 0.85 ± 0.02; P < 0.05) under RT-PCR and Western blot analysis. Meanwhile, the wounded area filled rate and the number of invaded cells of A549-siNanog cell group (57% ± 0.04, 69.60 ± 17.14) were decreased significantly compared to A549-siNC cell group (95% ± 0.02, 209.60 ± 15.40; P < 0.05). Conclusion siRNA targeting human Nanog could specially suppress the expression of Nanog gene in lung adenocarcinoma A549 cells. In this way, it couldsignificantly reduce the capability of migration and invasion of A549 cells.

Objective To evaluate the efficacy and safety of enoxacin in the treatment of lower respiratory tract bacterial infection in the old patients.Methods 48 cases were given enoxacin and 46 cefotaxime.Results There was no significant difference between the two groups in cure rate,effective rate,bacterial clearance and side effects incidence.Conclusion Enoxacin is effective and safe in the treatment of lower respiratory tract bacterial infection of the old patients.

Objective The evaluate action of serum L-arginine levels for predicting development of pregnant women with PIH,and analyze its effectiveness as clinical predictor.Methods Collecting 186 patients was performed a retrospective study.The PIH pregnant women was the experiment group,and the health pregnant women was the control group.In order to analyse the effect,the levels of L-arginine were measured in the early,middle and late period of pregnant women.Compared with the serum L-arginine in different groups with x2-test and t-test based on data type.Results Experimental group and control group subjects in age (x2 =2.426,P=0.119;t=1.218,P=0.229) and smoking before pregnancy (x2 =2.088,P=2.088),there was no significant differences,but the two groups of patients with BMI (x2 =8.772,P=0.003) and parity (x2 =6.083,P=0.014) was statistically.In different stages of pregnancy,the concentration of serum L-arginine had no statistical differences,and in the concentration of umbilical cord blood,serum L-arginine also had no statistical difference.There was a statistical differences in the serum L-arginine concentration of the cord blood for different number of pregnancies,but the concentrations of L-arginine in cord blood and in serum L-arginine.There were no significant difference in age,BMI and smoking pregnant.According to ROC curve analysis,for the diagnostic of concentration in serum L-arginine and in umbilical cord blood,the results indicate low efficiency to the diagnosis of pregnancy hypertension during pregnancy.Conclusion L-arginine level and the development of PIH and body mass index and maternal correlation for the pregnant women.Because the sample size limitations,L-arginine in the diagnosis of PIH also needs to be further research to determine the effectiveness of predicting.

ObjectiveTo study the cognitive function and observe the changes of event-related potential of epileptic children.Methods45 epileptic children and 45 normal children were put in as the test group and the control group.The cognitive function and event-related potential were evaluated by means of Raven's standard Progressive Matrices (SPM) and evoked potential instrument respectively.ResultsAbout 33.3% epileptic children presented cognitive deficits.For children with cognitive deficits, their scores of B, E were significantly different from those in the control group(P<0.05).The latency of event-related potential P300 showed significantly prolonged than that of the control group (P<0.01).ConclusionEpilepsy may cause cognitive function deficit, especially in the ability of analog, analysis and abstract. The latency of event-related potential P300 is a very good objective to assess the cognitive function of children.

BACKGROUND: Beside direct trauma, a series of secondary pathological changes would occur in local injured spinal cord area during spinal cord injury. It has been reported that recombinant human erythropoietin (rhEPO)could inhibit cell apoptosis and inflammatory reaction, and possess neuroprotective role.OBJECTIVE: To explore the neuroprotective role of rhEPO in spinal cord injury by observing the nerve cell apoptosis and related cytokine expression in traumatic spinal cord.DESIGN: Randomized and controlled study.SETTING: Orthopedic Surgery Laboratory of Affiliated Cooperation Hospital of Tongji Medical College, and Department of Pathology of Tongji Medical College, Central China Science and Technology University.PARTICIPANTS: The study was conduced at Orthopedic Surgery Laboratory of Affiliated Cooperation Hospital of Tongji Medical College, Central China Science and Technology University and Department of Pathology of Tongji Medical College from September 2003 to May 2004. Thirty healthy female adult rats were randomly divided into 4 groups: ① Six rats in blank control group only subjected to spinal cord exposure without injury. ②Eight rats in injury group were subjected to spinal cord injury without medication. ③ Eight rats in medication A group were treated with rhEPO.④ Eight rats in medication B group were treated with thEPO and βaeacine sodium.METHODS: ① Animal model preparation: Spinal cord injury model was established on 24 rats by using improved Allen method. ② Administration: Rats in medication A group were treated with rhEPO in dosage of 300 U/(kg·d) at postoperative 1, 3, 5, 8, 11 days, while rats in medication B group were given additional β-aeacine sodium in dosage of 0.1 mg/kg,once a day for consecutive 11 days. Rats in blank control group and injury group were injected with the same volume physical saline from tail veins.③ Neurological function: The neurological function was scored by the same examiner at 1 day and 12 days after model establishment. Behavioral observation: Basing on improved Gale's neurological functional behavioral analysis, 0 score presented severer dysfunction and 6 scores represents normal. Slope test: The gradient was determined by the grasping capability of rat, which could reflect the recovery of neurological function. ④ Pathological examination: Rat was put to death at postoperative 12 days; traumatic spinal cord was made into slices for HE staining. ⑤ Expression of apoptosis cytokine bcl-2, bax and fas in nerve cells: Specimen was collected for IHC stain and brown colored positive cells was counted for calculating positive expressing rate. ⑥ Apoptosis nerve cells: In situ end-labeling techniques was used to calculate apoptosis index (the number of apoptosis nucleus/total number). All data were analyzed by using paired chi-test.MAIN OUTCOME MEASURES: ① Neurological functional behavioral score and results of slope test. ② Histological observation of traumatic spinal cord. ③ Expression of apoptosis nerve cell cytokines. ④ Examination of apoptostic nerve cells.RESULTS: ① Neurological functional behavioral score and results of slope test: There was no significant difference between 1 day and 12 days in blank control group, while the gradient in traumatic group was significantly larger in 12 days than in 1 day (P ＜ 0.05). In both therapeutic A and B groups, the behavioral scores and slope gradient were found significantly larger at 12 days than 1 day (P ＜ 0.05) and that of traumatic group (P ＜ 0.05). ② Histological observation of traumatic spinal cord: Traumatic spinal cord became slim with a majority of necrosis and glial cell hyperplasia in it; most of neurological tissues were found normal in medication A and B groups, with the neural structure of medication B group better than A group. ③Expression of nerve cell apoptosis cytokines: bax and fas positive cells in traumatic group were more than medication group A and B [traumatic group of (25.75±3.37)% and (41.37±2.83)% vs medication group A of (19.87±3.56) and (26.00±3.29)% vs medication group B of (12.00±2.97)and (17.50±2.20)％, P ＜ 0.05]; bcl-2 was significantly lower in medication group A and group B [(9.75±1.83)％, (14.63±2.83)％, (21.63±5.34)％,P ＜ 0.05]. ④ Examination of apoptotic nerve cells: Cell apeptosis index in traumatic group was significantly higher than medication group A and group B (50.75±5.39, 34.75±3.01, 24.00±3.46, P ＜ 0.05).CONCLUSION: Cell apoptosis is an important kind of neuronal death following spinal cord injury. rhEPO can inhibit nerve cell apoptosis and possess neuroprotective effect for traumatic spinal cord.

To modify the surface property of poly lactide-co-glycolide (PLGA) by biomimetic mineralization to construct a new kind of artificial bone. PLGA films and 3-diamensional (3-D) porous scaffolds hydrolyzed in alkaline solution were minerilized in SBF for 14 days. The morphology and composition of the mineral grown on PLGA were analyzed with SEM, FTIR and XRD. The porosity of the scaffolds was detected by using the liquid displacement method. The compressive strength of the scaffolds was detected by using a Shimadzu universal mechanic tester. An obvious mineral coating was detected on the surface of films and scaffolds. The main component of the mineral was carbonated hydroxyapatite (HA) similar to the major mineral component of bone tissues. The porosity of the un-mineralized and mineralized porous scaffolds was (84.86 +/- 8.52) % and (79.70 +/- 7.70) % respectively. The compressive strength was 0.784 +/- 0.156 N/mm2 in un-mineralized 3-D porous PLGA and 0.858 +/- 0.145 N/mm2 in mineralized 3-D porous PLGA. There were no significant differences between the mineralized and un-mineralized scaffolds (P > 0.05) in porosity and biomechanics. Biomimetic mineralization is a suitable method to construct artificial bone.
Biocompatible Materials ; Bone Substitutes ; Calcification, Physiologic ; Durapatite/metabolism ; Lactic Acid/*chemistry ; Polyglycolic Acid/*chemistry ; Polymers/*chemistry ; Porosity ; Tissue Engineering

ObjectiveTo observe the expression of apoptosis factor, the mechanism of neuronal apoptosis and profective effect of recombinant Human Erythropoietin (rHu EPO) after spinal cord injury (SCI) in SD rats.Methods30 SD rats were divided into four groups including control group, SCI group, treatment groups A and B.The animal SCI model was established with Allen's method. The changes of nerve functions of rats of 4 groups were observed before and after treating with rHu EPO. The expressions of apoptosis factors (Bcl-2,Bax,Fas) were tested with immunocytochemistry technique, and apoptosis neurones were labeled with TUNEL dyeing.ResultsThe grade of nerve function was improved distinctly in treatment groups, but group B was better than group A.The number of the positive cells for Fas and Bax in SCI group was more than that in treatment groups (P<0.05), and group A was more than group B. The number of Bcl 2 positive cells in the treatment groups was greater than that in SCI group (P<0.05).ConclusionApoptosis is a important death mode of neuron after SCI. rHu EPO can distinctly improve the comeback of the nerve function and protect the nerve tissue.

Objective To study the expression of Livin and its relalionship with expression of p53 in small cell hmg cancer(SCLC).Methods Immunohistochemical S-P method was used to detect the ex-pression of Livin and p53 protein in 30 SCLC tissues and 16 para-cancerous lung tissues.Results Livin protein was expressed in 17 of 30 SCLC tissues(56.67%),but Livin protein showed low levels in para-can-cerous lung tissues(12.50%),P＜0.01.There was no significant correlalion between positive Livin protein expression and age,sex,TNM staging,lymph node metastasis and lumor diameter(P＞0.05).p53 protein was expressed in 14 of 30 SCLC tissues(46.67%),but p53 protein was no expressed in para-caneerous lung tissues,P＜0.01.The expression of Livin protein was positively related to the expression of p53 protein(P＜0.01).Conclusions The aberrant expression of Livin may be a new target for diagnosis and gene treatment of SCLC.The aberrant expression of Livin and p53 may play synergetic role in process of carcinogenesis of SCLC.

To study the expression of neurocyte apoptosis and the changes of caspase-3 and Fas after spinal cord injury (SCI) in rats, improved Allen's method was used to make model of acute SCI at the level of T9 and T10. The animals were divided into six groups: a control group and 5 injury groups. The segments of injured spinal cords were taken 6, 24, 48 h and 7, 15 days after injury for morphological studies, including HE staining, Hoechst33258 staining and TUNEL labeling. The expression of caspase-3 was detected by immunohistochemical staining and RT-PCR. TUNEL-positive cells began to appear in the compression region 6 h after the injury, mostly located in the gray matter. TUNEL-positive cells were found in both gray and white matter, reaching a peak at the 3rd day. They began to decrease at the 7th day, distributed mostly in the white matter. Fas increased at the 6th h and peaked at the 3th day. Caspase-3 mRNA increased at the 6th h, peaking 48 h after the trauma, and decreased after 7 days. The protein expression of caspase-3, as revealed by immunohistochemical staining, was similar to TUNEL in time. It is concluded that apoptosis takes place after spinal cord injury, and caspase-3 mRNA and protein expressions were enhanced in the apoptosis. The expression of caspase-3 has a positive correlation with Fas expression.