Objective To investigate the changes of serum and urine fluorion organic fluoride poisoning by inhaling, and to probe into the clinical application value of concentrations in different degrees and at different time in patients with acute evaluating the sertm and urine fluorion concentration in acute organic fluoride poisoning by inhaling. Method A study was carried out in 23 patients, who suffered from acute organic fluoride poisoning by inhaling and were admitted Zhejiang Quhua Hospital, from December 2000 to December 2008. According to the occupational acute organic fluoride poisoning diagnostic criteria(GBZ66-2002),23 patients were divided into mild poisoning group,moderate poisoning group and severe poisoning group. Serum and urine fluorion concentration of patients at 1,2,3,4,5 d after poisoning were measured by using Ion-Selective Electrodes. Fluorion concentration of 10 staffs of Fluorine chemical company was also measured at the same period as the control group. The values of serum and urine fluorion concentration were analyzed. Differences in serum and urine fluorion concentration between groups at different time points were compared by repeated measures ANOVA and variability were deemed as statistical significance when P ＜ 0.05. Results Compared with mild poisoning group, there was no statistically significant difference ( P ＞ 0.05) in serum and urine fluorion concentration at the same time point in 1 to 5 days after poisoning in moderate poisoning group, but there was statistically significant differences ( P ＜ 0.05 or P ＜0.01) in severe poisoning group. Compared with moderate poisoning group, there was statistically significant difference ( P ＜ 0.05) in serum and urine fluorion concentration at the same time point in 1 to 5 days after poisoning in severe poisoning group. Serum fluorion concentration in 1 to 5 days after poisoning in each poisoning groupswere statistically higher than those in control group ( P ＜ 0.05), but there was statistically significant elevation ( P＜ 0.05) in urine fluorion concentration only in 1 day in mild poisoning group, in 1 to 3 days in moderate poisoning groups, in 1 to 5 days in severe poisoning group. Conclusions Serum fluorion concentration can be used as the severity index of diagnosis and determine the extent in 5 days after acute organic fluoride poisoning by inhaling,and urine fluorion concentration can also be used as diagnostic indicators of intoxication, but only in earlier stage or severe poisoning.

Objective To investigate the effects of sevoflurane postconditioning on intestinal ischemiareperfusion (I/R) injury in rats.Methods Thirty-six adult male Sprague-Dawley rats,weighing 200-220 g,were randomly divided into 4 groups (n =9 each):sham operation group (group Sham),group I/R,ischemic postconditioning group (group Ipo) and sevoflurane postconditioning group (group Sevo).Intestinal I/R was induced by clamping the superior mesenteric artery (SMA) for 60 min followed by 120 min of reperfusion in groups I/R,Ipo and Sevo.In group Ipo the animals were subjected to 3 cycles of 30 min reperfusion-30 min ischemia starting from the beginning of reperfusion.The animals inhaled 1.15％ sevoflurane for 30 min starting from the beginning of reperfusion in group Sevo.The animals were sacrificed at 120 min of reperfusion and then the small intestines were removed for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity (by colorimetric method) and caspase-3 protein expression in intestinal tissues (by Western blot).The density of apoptotic cells was calculated by TUNEL.Results Compared with group Sham,the intestinal injury score,density of apoptotic cells and MDA content were significantly increased,SOD activity was decreased,and caspase-3 protein expression was up-regulated in groups I/R,Ipo and Sevo (P ＜ 0.05).Compared with group l/R,the intestinal injury score,density of apoptotic cells and MDA content were significantly decreased,SOD activity was increased,and caspase-3 protein expression was down-regulated in groups Ipo and Sevo (P ＜ 0.05).There was no significant difference in the intestinal injury score,density of apoptotic cells,SOD activity,MDA content and caspase-3 protein expression between Sevo and Ipo groups (P ＞ 0.05).Conclusion Sevoflurane postconditioning can attenuate intestinal I/R injury through reducing lipid peroxidation and cell apoptosis in rats,and the protective effect is similar to that of ischemic post-conditioning.

BACKGROUND:Schwann cells transplantation can change the local micro-environment and help to repair the injured neural tissue, so getting a large number of highly purified and active Schwann cells is the key of the study. OBJECTIVE: To search for a simple and rapid method to extract and purify the Schwann cells.METHODS: Rats were divided randomly into two groups, namely, in vivo pre-degeneration of sciatic nerve resection group and untreated control group, with 20 rats in each group. Under sterile conditions, the rat sciatic nerves were cut off at post-operative 7 days, Schwann cells were extracted by using mixed enzyme digestion and tissue mass transplantation; through low enzyme digestion and twice inoculation to differential adhesion, Schwann cells were purified. Cell morphology was observed under phase contrast microscope and identified by mmunofluorescence staining; cell purity was calculated; MTT method assay was used to determine the capacity of cell proliferation.RESULTS AND CONCLUSION: At 7 days of the culture, the experimental group showed the typical bipolar or bipolar Schwann cells, with connections between cells; in control group, cell processes were shorter and less associated with the surrounding cells. Following S-100 immunofluorescence staining, cells were positive for green expression.Cells proliferated rapidly in the experimental group and formed a swirling shape at 15 days, there were a relatively small number of fibroblasts, at the purity of 96.1%; in the control group, the cells proliferated slowly, with many fibroblasts at a low purity. MTT assay showed that primary cultured Schwann cell proliferated weakly in both groups; compared with the control group, the proliferation of subcultured Schwann cells in the experimental group was markedly increased (P < 0.05 or 0.01), and reached a peak 3 4 days later. The results confirmed that in vivo denaturing, in vitro hybrid enzyme digestion, tissue mass transplantation combined with low enzyme digestion, separation of double-differential adhesion of Schwann cells is a simple and rapid method to extract and purify Schwann cells.

BACKGROUND: Schwann cells are the seed cells of neural repair, and it is a key to harvest a large number of Schwann cells with high purity and activity. OBJECTIVE: To compare the in vitro culture, purification, and morphology of Schwann cells between neonatal and adult rats, and investigate a simple and feasible culture method to harvest high-purity Schwann cells. METHODS: Totally 30 Sprague-Dawley rats, comprising 20 neonatal (1-3 days after birth, neonatal group) and 10 adult (weighing 150-200 g, adult group) rats, were included. Following double-enzyme digestion and two incubations, Schwann cells were isolated and purified by differential attachment. Cell morphology and attaching speed were determined through the use of inverted microscope. Cells were counted and cell purity was calculated. Cell proliferative ability was detected by MTT microcolorimetry. Curves of cell proliferation in each group were depicted to determine proliferative speed. Schwann cells were identified by S-100 immunochemistry.RESULTS AND CONCLUSION: Compared with fibroblasts, neonatal rat Schwann cells exhibited faster, while adult rat Schwann cells showed slower, attaching speed. Both neonatal and adult groups yielded over 96% cell purity. MTT microcolorimetry results revealed that Schwann cells proliferated actively in neonatal and adult groups. Cell proliferative curves show that neonatal rat Schwann cells proliferated faster than adult rat Schwann cells (P < 0.05). S-100 immunochemistry results showed positive results in both groups. All these findings suggest that double-enzyme digestion and two incubations followed by differential attachment is a satisfactory method to harvest considerable Schwann cells with high purity and activity. Neonatal rat Schwann cells show stronger proliferative, attaching capacities than adult rat Schwann cells.

ObjectiveTo introduce a simple and quick method of adherent monoculture to primary neural stem cells depending on the mainly peptic characteristic of collagenase. MethodsThe fetus rat telencephalons of a pregnant 14-day-old SD rat were isolated, and tissues were treated with collagenase type Ⅰ contained EDTA following trituration. The cells were raised in polylysine culture plates with serum-free medium, and assessed with Nestin immunofluorescence. ResultsThe Nestin positve cells were obtained, and the purity was over 99%. ConclusionThis method may simply and quickly yields the primary neural stem cells that are monolayer and adherent.

Objective To study the expression of BCL10 and associated chromosomal aberration in primary cutaneous marginal zone B-cell lymphoma (PCMZL). Methods Tissue specimens were collected from 17 patients with PCMZL. Immunohistochemistry was used to detect the expression of BCL10. Fluorescence in situ hybridization (FISH) was performed to examine the presence of API2-MALT1 fusion gene and chromosomal aberration in BCL10, MALT1 as well as IgH genes in these cases. Results Of these patients,94.1% (16/17) expressed BCL10 protein. The cytoplasmic expression of BCL10 was observed in 64.7% (11/17) of the patients, and nuclear expression in 29.4% (5/17). As shown by FISH test, neither API2-MALT1 fusion gene nor chromosomal aberration in BCL10, MALT1 or IgH genes was present in these patients. Conclusions Compared with MALT lymphomas originating from tissues other than skin, PCMZL is uncommonly associated with chromosomal abnormalities; it is possible that there are unknown factors contributing to its tumorigenesis. Nuclear BCL10 is unrelated to the presence of chromosomal aberration in BCL10, MALT1 or IgH genes. Further follow-up is required to clarify the association between nucle ar BCL10 and poor prognosis of PCMZL.

BACKGROUND:Studies have shown that Schwann cells form a Bunger band in the basement tube and guide the extension of regenerating axons after peripheral nerve injury, but the exact mechanism remains to be explored.
OBJECTIVE:To explore the effect of Wal erian degeneration on biological characteristics and secretory function of Schwann cells in rats with sciatic nerve injury.
METHODS:A rat model of sciatic nerve injury was established and divided into two groups:sciatic nerve transection group and surgical control group. Schwann cells were isolated and cultured from sciatic nerve segments by one enzyme digestion. The cellmorphology was observed under light microscope and S-100 protein expression was determined by immunofluorescence staining. After subculture, the first generation of Schwann cells were chosen to draw the growth curve by the counting method within 14 days. The cellactivity was detected by MTT assay. The adhesion of Schwann cells was examined by acid phosphatase analysis and the concentration of nerve growth factor was detected by ELISA method.
RESULTS AND CONCLUSION:At 14 days after primary culture, a great number of Schwann cells were observed near the edges of nerve segments in the sciatic nerve transection group, but only smal number of Schwann cells scattered around nerve segments in the control group. Schwann cells in both groups showed S-100 positive expression. At 3 days after subculture, Schwann cells reached the logarithm proliferative phase, the cellnumber and proliferation absorbance values in both groups were increased along with time extension. Furthermore, the number of Schwann cells and absorbance value in the sciatic nerve transection group were significantly higher than those of control group (P<0.05). The adhesion ability in the sciatic nerve transection group was also significantly higher than those in the control group (P<0.05). ELISA results showed that, the concentrations of nerve growth factor in the sciatic nerve transection group were significantly higher than those in the control group at 4, 6, 8, 10, 12 and 14 days (P<0.05). After sciatic nerve injury, Wal erian degeneration can induce Schwann cells dedifferentiate into the precursors, significantly influence the biological function of Schwann cells, promote the proliferation of Schwann cells within the short term, secrete large amounts of neurotrophic factors, enhance celladhesion, and provide a suitable microenvironment for regenerated axons. In addition, it creates the necessary microenvironment for peripheral nerve regeneration.

Objective To observe morphological characteristics and activity distribution of T. asahii biofilm. Methods The morphological characteristics of T. asahii biofilm were observed under an inverted microscope and scanning electron microscope, and activity was measured and quantitatively analyzed by 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazo-lium hydroxide (XTT) assay and viable count, respectively. Spatial distribution of dead/vital cells, activity and thickness of biofilm at different layers were assessed under a confocal laser scanning microscope (CLSM) following double staining with FDA/PI. Results T. asahii formed a biofilm in vitro on the surface of polystyrene materials. Under a scanning microscope, the biofilm displayed a complex three-dimensional structure which composed of spores, pseudohy-pha and true hypha. As time prolonged, the activity and quantity of biofilm increased. The results of XTT assay were correlated with those of viable count (r = 0.94, P < 0.01). The activity was of no obvious difference between different layers of the biofilm. The thickness of biofilm varied from 14.3 μm to 31 μm. Conclusions The structure of T. asahii biofilm in vitro is more complex than that of planktonic T. asahii. The activity is of no significant difference between different layers of T. asahii biofilm.

Objective To investigate the CT features and different risk CT findings of intestinal stromal tumor. Methods The CT imaging data of 25 cases of intestinal stromal tumor confirmed by pathology and compared with operative and pathologic findings were retrospectively studied. Analyzing the CT features based on Histopathologieal classification of the different risk groups and using chi-square test to compare the differences. Results There were 9 cases which tumors originated from the jejunum, and 13 cases from ileum, only 3 cases from duodenum. Among them, 2 cases were submucosal type, 13 cases were intramural type, and 10 cases were subserous type. The pathologic patterns of different risk which included high-risk, intermediated-risk, low-risk, and very low-risk were 12 cases, 7 cases, 5 cases and 1 cases respectively. A typical CT manifestations of intestinal stromal tumors were a outward growth of irregular or round soft tissue mass originated in small intestine which had clear boundary and the non-homogeneous density, which corresponding to necrosis, cystic change, mucoid degeneration and sinus or cavity. Mesenteric fat invaded by tumor showed high-density lines or points shape. The Enhancement of lesion was obvious and not homogeneous which showed little change in peak of enhancement between arterial phase and venous phase. Intestinal stromal tumors took 5 cm as the boundary which including different size, shape, density, and appearance vessel-like artery shadow at arterial phase between different risk groups(low, very low-risk group and intermediated, high-risk group) were statistically different (P ＜0.05), while enhanced degree without significant difference (P ＞ 0.05). Conclusion CT findings of small intestine stromal tumor have characteristics and CT features have significant difference between different risk groups. These features of more than 5 cm in diameter, non-homogeneous density, irregular shape and chaos appearance like vascular enhancement are showed in intermediated-high-risk group.