Objective　To explore the phenomenon of neural apoptosis after chronic compressive spinal cord injury. 　Methods　A newly designed chronic progressive compressive spinal cord injury model of rat was adopted in this study. Forty-five Wister rats were divided into mild, moderate, and severe group according to the compressive degree of the spinal cord. The Feulgen stain and TUNEL were used to investigate the apoptosis and its characteristics in different kinds of neural cells. 　 Results　Apoptosis index in moderate injury group was the highest. Apoptotic cells largely located in ventral, lateral and dorsal column of the white matter. Most of them were oligodendrocytes. Positive neuron occasionally presented in laminae Ⅲ～Ⅸ part of the sections. Most of them located in dorsal gray horn. 　Conclusions　 Apoptosis is an important event in secondary pathophysiological process of chronic progressive compressive spinal cord injury. Apoptosis is one of the reasons for neural cell death. The apoptosis of oligodendrocytos may contribute to myelin sheath disruption of white matter.

Objective To investigate the effects of nimesulide, a selective cyclooxygenase-2 (COX-2) inhibitor, on human cholangiocarcinoma QBC939 cell line in vitro. Methods The effects of nimesulide on QBC939 cells were observed with the following techniques: the influence of nimesulide on the proliferation of QBC939 cells was determined by MTT assay; the apoptosis of QBC939 cells was viewed and measured by transmission electron microscopy and flow cytometry, respectively; the expressions of proliferation cell nuclear antigen (PCNA) and COX-2 of cholangiocarcinoma cells were detected by immunocytochemistry. Results Nimesulide inhibited the expressions of PCNA and COX-2 and the proliferation of cholangiocarcinoma QBC939 cells, whose effects intensified as the dose increased and time elongated. Flow cytometry showed that the apoptotic rates of QBC939 cells increased significantly as the dose of nimesulide increased. The typical morphologic features of apoptosis were also observed by transmission electron microscopy. Conclusion Nimesulide significantly inhibits the proliferation of QBC939 cells in vitro by inducting cell apoptosis, which may be associated with the downregulation of COX-2 expression, and it also presents the features of dose and time dependents.

Objective To evaluate the feasibility, safety and efficacy of selective portal vein embolization (SPVE) in treatment of liver tumor in rats and to provide the groundwork for its future clinical applications. Methods 24 healthy rats underwent the embolization. Pre and post SPVE portogram and liver chemical profile were obtained. Four rats were sacrificed at 10 min, 7,14, 21 and 28 days respectively following follow up portography. The liver, heart, lungs and kidneys were examined macroscopically and microscopically. Fifteen rats implanted with Walker 256 tumor sized from 3 to 10 mm in liver were scanned with MRI and portography pre SPVE taken. Post SPVE 3 rats were examined with MRI for each group at the same interval as above and the lives were examined microscopically. Results (1) The blood flow to the target portal branches were immediately halted after SPVE. These vessels remained occluded without collateral formation up to 28 days. (2) The liver indexes and BUN level increased after embolization, but returned to normal within 21 d. Macroscopic and microscopic changes were not found in the heart, lungs or kidneys. (3) In the healthy rats, the affected segment was atrophic and the remaining liver underwent compensatory hypertrophy. Histologic examination revealed that the targeted portal veins were coagulated, the endothelium were degenerated and the local hepatocytes were necrotic after embolization. (4) In the rats with implanted liver tumor, the affected segment including the tumor was necrotic and atrophic. The tumors were completely necrotic, and no viable tumor cell was seen under microscope in 12 among the 15 rats. Three tumors 10 mm in diameter were not completely necrotic. Part of tumor cells were still alive and infiltrated into the surrounding liver. Conclusion SPVE with ethanol is effective in the treatment of small liver tumor in rats. However,in case of bigger tumors involving several segments, SPVE should be combined with other treatment.

CD40 Ligand ; pharmacology ; HL-60 Cells ; drug effects ; Humans ; fas Receptor ; metabolism

Objective To investigate the effect of a selective inhibitor of COX-2 nimesulide on growth and apoptosis of human cholangiocarcinoma cell line QBC939 in vitro. Methods MTT assay was used to determine the influence of nimesulide on the proliferation of QBC939 cells, apoptosis of QBC939 was measured by transmission electron microscopy and flow cytometry.Expression of apoptosis related genes mRNA and bcl-2 ,bax, survivin were detected by RT-PCR and immunocytochemistry. Results Nimesulide effects a dose-dependent and time-dependent growth inhibition on QBC939 cells. High concentration of nimesulide (200 ?mol/L) not only inhibits the growth of QBC939 cells but also induces apoptosis cell nuclear condensation and apoptotic bodies were seen by transmission electron microscopy. Immunocytochemistry and RT-PCR shows upregulation of bax and down regulation of bcl-2 and survivin. Conclusion Nimesulide significantly inhibits the proliferation of QBC939 in vitro by induction of apoptosis in a dose- and time- dependent manner.

Adult ; Carcinosarcoma ; pathology ; Female ; Heart Neoplasms ; pathology ; Humans ; Lung Neoplasms ; pathology ; Myocardium ; pathology

Objective To observe pathological process of intestinal epithelial cells subjected to ischemia,ischemia/reperfusion injury and inflammation simulated hypoxia/reoxygenation (H/R) and lipopolysaccharide (LPS) challenged human fetal normal colonic cell (FHC) line in vivo,and to observe the changes when the assaulted intestinal epithelial cells were treated with emodin,in order to explore the possible intervention targets of emodin.Methods Normoxia group:the FHC cells were cultured in 95％ air and 5％ CO2 at 37 ℃.Hypoxia (H) group:the cells were cultured with a mixed anaerobic gas of 1％ O2,5％ CO2 and 94％ N2 at 37 ℃ for 1,2,3,4 hours.H + LPS group:the cells were cultured in hypoxic condition as H group with simultaneous challenge of LPS (1 mg/L).H/R group:the cells were cultured in hypoxia for 3 hours followed by reoxygenation for 1,2,3 and 4 hours,respectively.H/R + LPS group:the cells were cultured in H/R as H/R group and LPS (1 mg/L) simultaneously.Emodin intervention group:the cells were cultured in H3 h/R2 h + LPS and emodin (20,40,60,80 μmol/L) simultaneously.The variation trends of phosphorylation nuclear factor-κB profilin-o (pIκB-α),phosphorylation NF-κBp65 (pNF-κBp65) and their downstream target gene cyclooxygenase-2 (COX-2),and hypoxia-inducible factor-1α (HIF-1 α) were determined by Western Blot.The morphological changes in intestinal epithelium in different groups were observed using light microscope.The effect of emodin on the proliferation of intestinal epithelial cell was measured by methyl thiazolyl tetrazolium (MTT) assay.Results ① H group:the expressions of pIκB-α,pNF-κBp65 and COX-2 were upregulated,peaking at H1 h (0.350 ± 0.018,1.083 ± 0.054,0.903 ± 0.045),and then they gradually lowered (F value was 3.011,7.247,5.754,P value was 0.013,0.000,0.005,respectively).The expression of HIF-1 α peaked at H3 h (1.511 ± 0.076),but there was no significant difference among different groups (F=1.881,P=0.062).H + LPS group:the expressions of pIκB-α,pNF-κBp65,COX-2,HIF-1α were increased with elongation of duration of hypoxia,and a maximal induction was observed at H3 h (0.504 ± 0.025,1.255 ± 0.063,0.812 ± 0.041,1.209 ± 0.075,F value was 2.683,8.774,9.765,2.432,and P value was 0.011,0.000,0.000,0.026,respectively).H/R group:with the prolonged duration of reoxygenation,the expressions of NF-κB signaling pathway proteins (pIκB-α,pNF-κBp65,COX-2) were decreased and dropped to nadir at H3 h/R4 h (0.712 ± 0.034,1.202 ± 0.048,0.691 ± 0.042,F value was 1.923,6.765,2.719,and P value was 0.063,0.000,0.016,respectively).Compared with H group,HIF-1α was decreased with a prolonged duration of reoxygenation in H/R group,but there was no significant difference in value among different time points (F=1.280,P=0.081).H/R + LPS group:pIκB-o,pNF-κBp65,COX-2,HIF-1α showed no sign of degradation with the prolonged duration of reoxygenation,and their expression increased to maximum analogously at R2-3 h (3.302 ± 0.061,2.315 ± 0.055,2.017 ± 0.043,2.413 ± 0.098,Fvalue was 4.614,1.652,5.970,2.076,and Pvalue was 0.001,0.067,0.000,0.037,respectively).Emodin group:emodin when co-treated with H/R + LPS inhibited the expression of HIF-1o and NF-κB pathways with a dose-effect relationship (P＜0.05 or P＜0.01).Emodin at the dose of 80 μmol/L showed most marked inhibition (2.599 ± 0.130,1.772 ± 0.089,2.590 ± 0.129,2.518 ± 0.125).However,after treatment of emodin did not show such effect.② After treatment with H/R + LPS,there were morphological changes in cells:vacuoles,deformation and fusion.The speed of cell growth became much slower compared with H group.③ Emodin (20-80 μmol/L) had no significant effect on cell proliferation.Although emodin produced biological effect in this concentration range,it had no cellular toxicity.Conclusions Both hypoxia and inflammation can activate the hypoxia pathway of HIF-1α and the pro-inflammatory pathway of NF-κB,but different stimuli cause varying degrees of activation in these two pathways.In H/R group,both pathways were weakened during reoxygenation.However,in H/R + LPS group,the proteins remained to show a relatively high expression during the process of reoxygenation.This may be related to the pathophysiological mechanism of intestinal ischemia/reperfusion injury:hypoxia/reperfusion injury and LPS act together to destroy the intestinal epithelial cells and induce gut-derived sepsis.Emodin may inhibit inflammation by blocking HIF-1α/NF-κB-COX-2 signaling pathways.

Objective To explore the clinical features of thoracolumbar disc herniations and to improve the quality of the diagnostic procedure.Methods Clinical data of 65 patients with thoracolumbar disc herniations confirmed by X-ray examinations,CT,MRI,and operations from September 1995 to January 2004 were retrospectively reviewed.The 65 patients were divided into three groups: lower thoracic disc herniations(T_(10-11)~T_(12)L_1) in 43 patients,upper lumbar disc herniations(L_(1-2)~L_(2-3)) in 16 patients,and multiple levels of herniations in 6 patients.Results Paresthesia and lower extremity weakness were the most frequent symptoms,with their occurrence proportions being 89.2%(58/65) and 83.1%(54/65),respectively.Among the 65 patients,9.2%(6/65) showed the presentation of upper motoneuron involvement,47.7%(31/65) manifested symptoms of lower motoneuron impairment,and 43.1%(28/65) presented as mixed motoneuron disorders.Neurological deficits were usually extensive and the cauda equino syndrome was commonly seen,while isolated radicular impairment was noticed only in 3 patients.Back pain(44.6%,29/65) and lower extremity weakness(40.0%,26/65) were the most common initial complaints.Lower thoracic disc herniations were characterized by mixed motoneuron disorders at the occurrence proportion of 58.1%(25/43),with a tendency leading to ambulatory dysfunction,drop foot,increased lower extremity muscle tension,and positive pathologic reflexes.By contrast,most upper lumbar disc herniations were manifested as lower motoneuron disorders at the occurrence proportion of 93.8%(15/16),with back pain,lower extremity pain,and the cauda equino syndrome frequently encountered.Conclusions The clinical presentation of thoracolumbar disc herniations is complicated with the large-scale distribution and diversity of the symptoms and the complexity of clinical signs.We put forward four circumstances under which a high suspicion of thoracolumbar disc herniation was recommended: ①if there is a sensory disturbance at the anterior and lateral aspect of the thigh or at the groin area;②if there is a lower extremity weakness,especially in the quadriceps and the tibialis anterior muscle(drop foot);③if an extensive and irregular range of sensory and motion disturbances exists,with a lack of typical radicular distribution;or ④if there are mixed motoneuron disorders,or lower motoneuron disorders unexplained by lumbar disc herniations.

Objective To prepare and identify monoclonal antibodies (mAb) against histidine-tag (His-tag ) .Methods Balb/c mice were immunized with polypeptides containing 15 histindine(His)-coupled BSA and this fusion was prepared according to con-ventional methods .Indirect ELISA was used to screen the positive clones and limited dilution for further cloning .After purified with Protein G affinity chromatography ,these antibodies for His-tag were detected for antibody titer ,relative affinity as well as subtypes by using ELISA .The specificity of these mAb was identified by ELISA and Western blotting analysis .Double-antibody sandwich ELISA was composed of these mAb paired with the corresponding recombinant protein of specific antibody ,which was used to de-tect the recombinant protein quantitatively .Results 5 hybridoma cell lines stably secreting anti-His-tag IgG1(κ) were screened .A-mong these antibodies ,1-G2 was of the highest affinity ,reaching 1 .27 × 1010 ,which could combine with different recombinant pro-teins containing His-tag in the experiment of ELISA and Western blotting .1-G2 also could be used to detect recombinant PCT pro-tein containing His-tag quantitatively by using double-antibody sandwich ELISA with antibody of PCT ,the sensitivity of detection reached 4 .6 ng/mL .Conclusion The mAb against for His-tag with high specificity ,affinity and secreted stably are successfully prepared .This prepared antibody could be used in ELISA and Western blotting to detect a variety of His-tag .

To study photosynthetic characteristics and its influencing factors in leaves of medicinal plant Mirabilis himalaica, and provide an evidence for guiding artificial planting and improving the quantity. The light-response and diurnal photosynthesis course of leaves at the booting stages of 1-3 year old M. himalaica were measured with LI-6400 system. The Results showed that the light response curves were fitted well by non rectangle hyperbola equation (R2 > or = 0.98). The values of the maximum photosynthetic rate (Pmax) and light use efficiency of three-year old M. himalaica leaves were higher than those of 1-2 year old individuals. The diurnal variation of net photosynthetic rate (Pn) and stomatal conductance (Gs) of 2-3 year old M. himalaica were typical double-peak curves determinately regulated by stomatal conductance. However, transpiration rate (Tr) of 1-3 year old plants leaves were single-peak curve, which was self-protection of harm reduction caused by the higher temperature at noontime. Correlation analysis showed that the changes between photosynthetic active radiation (PFD), air temperature (T ) and Pn, were significant positive related. Therefore, M. himalaica is a typical sun plant, which should be planted under the sufficient sunshine field and prolong the growing ages suitably in order to improve the yield.
Breeding ; Mirabilis ; growth & development ; metabolism ; Models, Biological ; Photosynthesis ; Temperature ; Time Factors