Objective To investigate the role of NF-kB in radiation-induced apoptosis ot human non-Hodgkin lymphoma (NHL) cells and the mechanism involved.Methods Three human non-Hodgkin lymphoma cell lines,Namalwa,Ramos,and Raji cells were divided into control,IR and IR + QNZ ( 10nmol/L) groups,respectively.Annexin-V kit was used to determine cell apoptosis. Protein expression levels of Survivin,Bax,Bcl-2 and cleaved Caspase-3 were evaluated by Western blot.Survivin mRNA was quantified by real-time PCR.Results Inhibition of NF-kB by QNZ pretreatment significantly enhanced X-ray induced apoptosis in human NHL cells in a dose-dependent manner (t =2.93 - 12.52,P ＜ 0.05).At the same time,QNZ significantly reversed the expression levels of Survivin protein and mRNA that were upregulated by radiation(t =3.29 - 16.72,P ＜ 0.05).QNZ also increased the levels of Caspase-3 and proapoptotic protein Bax,but reduced anti-apoptotic protein Bcl-2 expression level and hence the ratio of Bcl-2/Bax.Conclusions Inhibition of NF-kB could enhance radiation-induced cell apoptosis in human NHL cells through down-regulating Survivin expression and decreasing Bcl-2/Bax ratio.

BACKGROUND: Astragalus root can inhibit apoptosis through reducing the release and interstitial accumulation of excitatory amino acids, alleviating calcium overloading and antioxidative effect.OBJECTIVE: Astragalus root was used to treat anoxic-ischemic brain injury in immature brain. We evaluated the effect of astragalus root on caspase-3 mRNA expression, and meanwhile, labyrinth test was employed to investigate the intervention of astragalus root on learning and memory function of mature rats after anoxic-ischemic brain injury.DESIGN: Randomized and controlled study.SETTING: Pediatric Department, Zhongda Hospital Affiliated to the Medical College of Southeast University; Pathological Department, the Basic Medical Sciences Institute of Southeast University.MATERIALS: From October 2002 to June 2003, this study was conducted at the Experiment Center of the Medical College, Southeast University.A batch of 114 seven-day-old SD rats were selected from the same brood and divided into 3 groups, namely, sham-operation group (n=18), model group (n=48) and astragalus root group (n=48). Astragalus injection was produced by Chengdu DIAO Pharmaceutical Factory, with 10 mL astragalus injection corresponding to 20 g raw material.METHODS: Animal model of anoxic-ischemic brain injury was established in model group and astragalus root group, but was not established in sham-operation group. In astragalus root group, immediately after establishing anoxic-ischemic model and at the same time point each day, 0.08 mL astragalus injection was administered intraperitoneally until the 7th postoperative day. In model group, 0.08 mL normal saline was administered at the same time points. In sham-operation group, no treatment was given. In astragalus root group and model group, animals were decollatedat 24 hours and 5 days postoperatively to take out the brains. In sham-operation group,animals were decollated and their brains were taken out at 24 hours postoperatively. In all the groups, hippocampal brain injury was detected using histopathological method combined with semi-quantified RT-PCR methods for detecting caspase-3 mRNA. Adult rats aged 90 days were used in modified y maze to examine their learning and memory functions. All these three experiments were independent.MAIN OUTCOME MEASURES:① Hippocampal brain injury in each group was evaluated using pathological method.② Caspase-3 mRNA in the ligated side of hippocampus was detected.③ Results of modified Y maze test were analyzed.RESULTS:All of the 114 rats entered the statistical analysis.① Assessment ofhippocampal brain injury in each group with pathological method:In sham-operation group, the bilateral hippocampus showed no swelling or necrosis, and neural cells in this area had normal morphological features with a density of (87.7±0.6) × 103 per high amplification field. In model group, the ligated side of hippocampus was swollen with a widened spatium and the cell density decreased to (68.8±3.0) × 103 per high amplification field, which significantly differed from that in sham-operation group (P ＜ 0.01). At the fifth day, the volume of ligated side of hippocampus reduced with pyramid layer disorganized and neural cells sparse at a density of (48.7±2.2) × 103 per high amplification field. These changes were significantly different from those of sham-operation group and the same side at 24 hours (P ＜ 0.01). At 24 hours the ligated side of hippocampus was less swollen in astragalus root group than in model group.At day 5, the whole hippocampus was observed. At these two time points,cell death rate in astragalus root group was significant lower than that in model group(P ＜ 0.01).②Caspase-3 mRNA in the ligated side of hippocampus in all the groups: In sham-operation group, the expression of caspase-3 was low, with an absorbency value of 0.220±0.009. In model group, after ischemia and anoxia its expression increased. At 6 hours, it was 11% higher than that in sham-operation group. In astragalus root group, mRNA level reached its peak, which was 260% higher than that in sham-operation group (P ＜ 0.01). The peak of mRNA continued, decreased after 48 hours and returned to baseline at 5 days and 7 days. The fluctuation of mRNA was similar between astragalus root group and model group,but the peak value at 24 hours and 48 hours in astragalus root group was 44％-46％ lower than that in model group (P ＜ 0.01). ③ Results of modified Y maze test: As compared to model group, in astragalus root group, the number of training times for meeting the standard made by the Association was significantly smaller [(45.7±2.7), (16.1±2.5) times, P ＜ 0.01] and at 24 hours after anoxia and ischemia, memory retention was significantly higher [(48.3±11.7), (80.0±9.0)%, P ＜ 0.01].CONCLUSION: Astragalus root can effectively inhibit the apoptosis of neural cells in hippocampus in immature brain after anoxia and ischemia and enhance the survival rate of them. This protective effect may be related to its inhibitory effect on the expression of caspase-3. Meanwhile, astragalus root can dramatically improve learning and memory function of the immature brain after anoxia and ischemia.

Objective To investigate the expression and significance of hypoxia-inducible factor-1α (HIF-1α) in the retinal neovascularization by metabolic acidosis in newborn rats. Methods One hundred and twenty newborn SD rats were randomly divided into acidosis (experiment) and normoxia (control) groups. A total of 60 newborn rats in experiment group underwent tubal feeding day for 6 days and followed by a period of recovery. The rats in the two groups were sacrificed at the 3rd, 5th, 8th, 10th, 13th and 20th day after birth, respectively. The morphologic changes of retinal vessels were estimated by observing the vascular pattern in adenosine diphosphatase stained retina flat mounts. The newborn vessels were quantified by HE staining. Immunohistochemical method was used to detect HIF-1α expression. Results In experiment group, numerous neovascularization and un-perfused area at the periphery of vessels occurred on the 10th day. The result of HE staining showed that in experiment group of 10-day old,the number of neovascular nuclei extending into the vireo was 28.78±7.53, and that of the control group was 1.22±1.48 (t=11.169,P<0.01). The results of immunohistochemistry revealed that the expression of HIF-1α protein were stronger in the experiment group than in the control group on the 8th, 10th and 13th day, and there were significant differences between the two groups (108.87±15.21, 183.68±26.58 and 129.42±9.85 vs 74.98±4.50, 76.38± 3.38 and 74.78±1.86, t=4.625, 9.023 and 9.672,P<0.05). Conclusions HIF-lα might play an important role in retinal neovascularization.

BACKGROUND:Currently, discectomy, fusion or decompression is considered an effective and conventional method for the treatment of spinal disease. Although there have been many reports on the adverse effects of diabetes on spinal surgery, but there are stil some differences. OBJECTIVE:To systematical y evaluate the observational studies and case-control studies about the effect of diabetes on the complications of spinal surgery. METHODS:The control ed and comparative studies regarding the effect of diabetes on the results and complications of spinal surgery were searched from the database according to the inclusion criteria. The observed indicators including mortality, revision rate, surgical site infection, the incidence of venous thrombosis, blood loss, operative time and hospitalization time. Two authors participated in extracting the data and evaluating the methodology and quality of the included studies. Meta-analysis was conducted according to the guidelines of epidemiological observational studies (MOOSE). The risk assessment of the extracted data was conducted using RevMan 5.2 software. RESULTS AND CONCLUSION:Eighteen literatures, involving 2 824 063 patients, were eventual y enrol ed. The experimental result showed that the mortality, surgical site infection, incidence of venous thrombosis of diabetic patients after the spinal surgery were significantly higher than those of non-diabetic patients;the hospital stay was significantly longer than that of non-diabetic patients (P<0.05). There were no significant differences in the risk of revision, intraoperative blood loss and operation time between diabetic patients and non-diabetic patients (P>0.05). These results suggest that diabetic patients take a higher risk once accepting the spinal surgery than the non-diabetic patients. Diabetes increases the risks of postoperative mortality, surgical site infection, venous thrombosis and hospitalization time after spinal surgery.

Objective:To investigate the neuroprotective effect and possible mechanism on rats with low dose Lipopoly -saccharide ( LPS) preconditioning after spinal cord injury.Methods:120 female SD rats were randomly divided into the empty virus (EV) group,LPS+empty virus (LPS+EV) group,Nrf2 interference virus (NIV) group,LPS+Nrf2 interference virus (LPS+NIV) group.The model of traumatic spinal cord injury ( TSCI) was established by the modified Allen′s method,motor function of the rat hind limb was assessed by the Basso Beattie and Bresnahan (BBB) score at 1,3,7,14 and 28 d after the operation.The injured spinal cord tissue samples were harvested at each time ,and the pathological changes of rat spinal cord were observed by HE staining ,the Nissl body and neuron survival index were observed by Nissl staining ,the expressions of Nrf2 and GCLC protein level were detected by immunohis-tochemical staining and Western blot.Results:The rat BBB score of LPS+EV group increased significantly than EV group at 7,14,28 d after operation ( P<0.05 ,P<0.01 );The NIV group between LPS+NIV group have no statistical significance at each time.As compared with EV group:the Nrf2 protein of LPS+EV group was expression increased significantly and Nissl staining showed that the neurons survival index was increased at 1,3 and 7 d(P<0.05,P<0.01);The GCLC protein of LPS+EV group was expression increased significantly at 1-14 d( P<0.05 );HE staining showed that the injured spinal cord pathological changes of LPS +EV group was obviously improved.Conclusion:Low dose lipopolysaccharide preconditioning can accelerate the nerve function recovery on rats with traumatic spinal cord injury ,the mechanism may be regulated by activating the Nrf 2 antioxidant stress pathway.

Objective:To assess the possible role of expression of TGFα and EGFR in nasal polyps and its re-lationship with PCNA labeling index. Method: Specimens from 20 patients of nasal polyps were studied with im-munohistochemical technique. Result: The expression of TGFα,EGFR and PCNA were increased in the epitheli-um, gland cells and inflammatory cells of nasal polyps. There was a close correlation between the intensities ofTGFα,EGFR and PCNA. Conclusion: TGFα may play a key role in epithelial cell proliferation in nasal polyps.

0.05). The valve competence recovery rate in severe PDVI group was significantly lower than that of the mild and moderate PDVI groups (both P

Background and purpose:Ovarian cancer is associated with a high recurrence and mortality due to the existence of cancer stem cells. The purpose of this study was to investigate the effect of casticin (CAS) on the capability of self-renewal in ovarian cancer stem cell like cells (OCSLCs) derived from SKOV3 cell line. Methods:SKOV3 cell line cells were cultured in vitro, and OCSLCs were obtained and amplified through suspended culture with conditioned medium of the stem cells. The phosphorylation level of FoxO3a was analyzed using Western blot. The protein expression of FoxO3a was inhibited by FoxO3a speciifc siRNA transfection, and then the ratio of sphere-formation was detected. Results: Compared with parental cells, OCSLCs over-expressed phosphorylated FoxO3a (pFoxO3a) and had elevated ratio of sphere-formation [(3.1±0.3)% vs (34.8±6.8)%, P<0.05]. CAS significantly inhibited the capability of sphere-formation in OCSLCs and down-regulated the expression level of pFoxO3a. And the transfection of FoxO3a speciifc siRNA suppressed the protein expression of FoxO3a and attenuated the inhibitory effect of CAS on the sphere-formation of OCSLCs. Conclusion: Reduced expression level of pFoxO3a is involved in the effect that CAS inhibits sphere-formation of OCSLCs derived from SKOV3 cell line.

Objective To study the value of the semiquantitative-parameter analysis of wash out index of time-intensity curve (Swash-out) in evaluating the therapeutic effect of neoadjuvant chemotherapy for locally advanced breast cancer (LABC).Methods Fifty-nine women with LABC underwent dynamic contrast enhancedt MRI examination before chemotherapy,after the 2nd cycle and the 4th cycle of chemotherapy.All patients were divided into major histological response group (MHR) and non-major histological response group (NMHR) according to the final pathologic response.Swash-out and the variancetrends of Swash-out before NAC,after the 2nd cycle of NAC and after the 4th cycle of NAC were compared in each group and between the two groups.According to the gold standard of Miller & Payne criterion,Receiver operating characteristic curve (ROC) analysis was performed to evaluate the predicting effect of Swash-out for NAC response,and to compare it with Semi-quantitative TIC curve indicators Smax (steepest slope) and PPE (peak percent enhancement).Results Fifty-nine patients of LABC patients were divided into a MHR group of 34 patients and a NMHR group of 25 patients.Swash before NAC of MHR group was-16.99 (-56.72-41.20),Swash-out after the 2nd cycle of NAC was 5.66(-69.45-53.08),Swash-out after 4th cycle of NAC was 15.95 (-7.80-54.23).Swash-out before NAC of NMHR group was-23.08 (-64.24-34.39),Swash-out after the 2nd cycle of NAC of NMHR group was-23.01 (-52.72-28.70),Swash-out after 4th cycle of NAC of NMHR group was-11.45 (-50.49-50.93).Swash-out variance rate of MHR group after the 2nd and the 4th cycle of NAC were-1.18 (-31.32-60.86) and 1.50 (-86.27-3.61),respectively.Swash-out variance rate of NMHR group after the 2nd and the 4th cycle of NAC were-0.28(-3.24-9.46) and 0.27 (-5.34-3.11),respectively.Swash-out was not significantly different between the two groups before NAC (Z =-0.97,P ＞0.05).Swash-out and Swash-out variance rate of MHR group after the 2nd cycle of NAC were significant higher than that of NMHR group (Z =-3.97 and-3.02,P ＜0.01).Swash-out and Swash-out variance rate of MHR group after the 4th cycle of NAC were significant higher than that of NMHR group (Z =-3.96 and-3.16,P ＜ 0.01).Area under curve (Az) after the 2nd and the 4th cycle of NAC were 0.805 and 0.804,respectively,and no significant difference was found between them (Z =0.019,P ＞0.05).Diagnostic cut-off points were-8.670 for the 2nd cycle of NAC and 4.105 for the 4th cycle of NAC.Diagnostic sensitivity was 79.42％,specificity was 76.00％ and Youden index was 0.554,for after the 2nd and the 4th cycle of NAC.Conclusion Swash-out of TIC curve before NAC cannot predict the response of NAC,Swash-out of TIC curve after the 2nd cycle of NAC and after the 4th cycle of NAC are efficient in predicting the response of NAC.

Objective To explore whether Forkhead O transcription factor-3a (FoxO3a) activity affects the capability of sphere-formation of ovarian cancer SKOV3 cell line.Methods Sphere-forming cells (SFCs) were obtained and amplified through suspended culture with conditioned medium of the stem cells in SKOV3 cell line.After SKOV3 cells were transfected with FoxO3a specific siRNA,the protein expressions of FoxO3a and Bmi1 and the ratio of sphere-formation were compared with Western blot and sphere-forming assay,respectively.Results Compared to parental cells,SFCs from SKOV3 cell line had higher ratio of sphere-formation and over-expressed Bmi1 and pFoxO3a.Transfection of FoxO3a specific siRNA down-regulated the protein expression of FoxO3a and upregulated expression of Bmi1 in SKOV3 cells,and enhanced the capability of sphere-formation.Conclusions Silence of FoxO3a leads to enhanced capability of sphere-formation in SKOV3 cell line.