Objective To investigate the differential expression of synaptophysin(SYN)in the developing rat cochlea,to analyze the relationship between the expression of SYN and auditory functional development,and to explore the source of the adenosine triphosphate(ATP)in the cochlea.Methods The immunofluorescence was applied to observe the differential expression of SYN in the cochlea of SD rats on different days(p1,p5,p10,p14 and p28)after parturition.Results No significant stain was observed in the Kolliker organ or the Corti's organ of P1,P5 and the top turn of the cochlea in P10.The SYN expression was turned on the outer spiral bundle,the inner spiral bundle,and the medial wall of the Deiters' cell of the cochlea in P14,P28 and the second or the third turn of Corti's organ of P10.All the spiral ganglion neurons(SGN)were stained in different aged cochlea.Conclusion The expression of SYN was different during development of rat cochlea.This difference was in favor of the configuration constructed between nerve endings and target cells.It also played a key role in the formation of correct auditory coding during auditory system development.The hair cells and the supporting cells,possible the source of ATP in the cochlea,contained ATP that maybe existed in a non-vesicular form or specific staining of non-SYN.

OBJECTIVE: To study the apoptosis/proliferation of Kölliker organ supporting cells and to understand the prompting apoptosis factors in vivo in the supporting cells in the Kölliker organ by changing the environment of the cultured supporting cells in the Kliker organ in vitro, via the separation, culture and purification of the supporting cells in the K6lliker organ. METHOD: A combinatorial approach of enzymatic digestion and mechanical separation was employed to isolate and culture in vitro pure Kölliker organ supporting cells. The purity was tested by flow cytometry assay. And K6lliker organ supporting cells were harvested to detect the rate and cycle of apoptosis by flow cytometry after Annexin V/PI staining, to test the cell growth curve by MTT assay, and to observe the differential expressions of the Bcl-2, Caspase-3, Caspase-8 and Caspase-9 through the Realtime PCR and Western blot. The calcium, potassium and glutamate concentrations in the culture medium of these cells in vitro were changed to detect the survival rate of cells by MTT assay. RESULT: The purity of K6lliker organ supporting cells by flow cytometry assay was 96. 56%. And these cells showed no significant difference in apoptosis, but an evident linear growth. The results of Realtime PCR and Western blot showed that the expression of Bcl-2, Caspase-3, Caspase-8 and Caspase-9 mRNA and protein in all different time points kept stable. Furthermore, the elevation of extracellular Ca2+ might contribute to decrease the cell viability of supporting cells. And K+ participated regulation of cell viability in a concentration-depending way. However, glutamate appeared to be a protective factor in high concentration. CONCLUSION There is no significant apoptosis in vitro of the supporting cells in the Kölliker organ of rats, showing a linear growth. The Ca2+ in high concentration might contribute to the apoptosis factor of these cells. However, the K+ and glutamate appear to be protective factors in high concentration.
Animals ; Animals, Newborn ; Apoptosis ; Caspase 3 ; Cell Cycle ; Cell Proliferation ; Cell Survival ; Cochlea ; cytology ; growth & development ; Flow Cytometry ; In Vitro Techniques ; Rats

Objective: To investigate the effects and mechanism of transhinone on calcium overloaded injuried models.Methods: Three injured models induced by caffeine, KCI, and NMDA, respectively, were used to assay the action of tanshinone in cultured primary cortex neurone of baby rats. Results: It was found that tanshinone possessed obvious protective effects on primary neurone in injured models by the way of morphological examination. Crystal violet staining and lactic dehydrogenase (LDH) measurement in supernate also indicated that tanshinone increased number of live neurone and reduced the extent of cell injury significantly.Conclusion: Tanshinone protected rat cortex cells from three kinds of calcium overloadied injured effectively in vitro.

Objective To explore the clinical features of nephrotic syndrome combined with H1N1 influenza. Methods The clinical manifestations, laboratory and image examinations, treatment, and prognosis of nephrotic syndrome combined H1N1 influenza were retrospectively analyzed in 15 children with. Results All of 15 children with nephrotic syndrome met the diagnostic criteria of H1N1 influenza. The median age of all children was 4-year-8-month old (2-year-2-month to 6-year-9-month). All children were treated with hormone alone or combined with other immunosuppressive drugs. Three cases were severe and another 5 cases were critically ill. Four cases were complicated with recurrence of nephrotic syndrome, 2 of which suffered from acute renal insufficiency. All children were given oseltamivir as antiviral treatment at admission. Four cases took oseltamivir within 48 hours of onset and showed mild symptoms. Fourteen children with H1N1I influenza were cured, their urinary proteins were significantly decreased or converted to negative, and the median hospital stay was 8 days (1 to 25 days). One child died of acute necrotizing encephalopathy and brain herniation. Conclusions Children with nephrotic syndrome are susceptible to severe or critical H1N1 influenza infections. During the epidemic of H1N1 influenza, the clinical preventive measures should be taken in children with nephrotic syndrome.

Adult ; Anemia, Aplastic ; chemically induced ; immunology ; therapy ; Benzene ; poisoning ; Cyclosporine ; therapeutic use ; Female ; Humans ; Immunity, Cellular ; Immunosuppressive Agents ; therapeutic use ; Male ; Middle Aged

Objective:To culture canine dental pulp stem cells(cDPSCs)in vitro.Methods:Canine pulp cells were isolated and cultured by enzyme digestion and explanted tissue culture respectively.Cell morphology was observed under phase-contrast micro-scope.The clone forming unit(CFU)of the cells was examined by plate clone formation assay.Cell markers and protein-expression were examined by flow cytometry(FC)and immunofluorescence.Odontogenic and adipogenic potential were evaluated by alizarin red staining and oil red O staining.Results:Short spindle fibroblast-like and steadily growing cells were obtained by both methods.The clone assay showed that CFU was 1 5.1 7% ±2.79%.FC observasion showed that the CD90,STRO-1 and CD24 positive cells were 24.43% ±7.1 0%,20.67% ±1 .42% and 2.03% ±0.06% respectively,but CD34 was negative.Immunofluorescence analysis showed positive expression of Nestin,Vimentin,weak expression of ALP and negative expression of DSP of the cells.Differentiation ex-periment confirmed the odontogenic and adipogenic differentiation potential of the cells.Conclusion:cDPSCs can be cultured in vitro.

BACKGROUND:Neural stem cel (NSC) transplantation is a common method for various ischemicencephalopathies, but inability to survive in the transplantation region limits its further use in clinical practice. OBJECTIVE:To explore the effect of vascular endothelial progenitor cel s (VEPCs) on the proliferation and apoptosis of co-cultured NSCs as wel as vascular remodeling in rats with ischemia reperfusion injury. METHODS:125 Sprague-Dawley rats were randomly divided into five groups, 25 rats in each group, including sham operation, ischemia, NSCs, co-culture, and VEPCs groups. Rat models of ischemia reperfusion injury were made in al groups except for the sham operation group, fol owed by corresponding interventions. The proliferation and apoptosis of neural stem cel s were detected, and vascular remolding in the ischemic region was observed in each group. RESULTS AND CONCLUSION:At different time points after transplantation, BrdU positive cel s were not observed in VEPCs, ischemia and sham operation groups;the number of BrdU positive cel s in the co-culture group was significantly higher than that in the NSCs group (P<0.05);BrdU+/Caspase-3+cel were observed in both co-culture and NSCs groups, and the apoptosis rate of the co-culture group was significantly lower than that in the NSCs group (P<0.05);there were new blood vessels in al the groups except for the sham operation group, and the number of new bone vessels was highest in the co-culture group. To conclude, our experimental results show that VEPCs promotes the proliferation of co-cultured NSCs, inhibits cel apoptosis and and promote angiogenesis in the ischemic penumbra of rats with ischemia reperfusion injury.

To study the level of blood selenium(Se)and nitrogen monoxide(NO)of oral lichen planus patients.Tested the contents of blood selenium(Se)and nitrogen monoxide(NO)of oral lichen planus patients and healthy cases.The level of blood selenium(Se)of oral lichen planus patients was lower than that of the control group,the situation of nitrogen monoxide(NO)was opposite,which indicated that Se and oxyradical may play an important role in the mechanism of oral lichen planus.

Objective To evaluate the significance of the panel of 6 microsatellites in detection of bladder cancer. Methods In the tumor tissue and urine sediment of 32 cases of bladder cancer 10 microsatellites were chosen and PCR-SSLP silver staining assay was conducted according to the methods described in the literature and our previous study.15 cases of non-bladder cancer served as controls. Results Microsatellite alternate (MA) was found in 30 out of 32 cases of bladder cancers,the sensitivity being 93.8%.The MA of urine sediment of 15 cases of non-bladder cancer was negative,the specifity being 100.0%.Among the 10 microsatellites, 6 ones were chosen;the MA positivity of the panel of the 6 ones was 90.6% (29/32).This result was not significantly different from that of the panel of 10 microsatellites. Conclusions MA assay is a sensitive,effective method for detection of bladder cancer.Compared with the panel of 10 microsatellites,the panel of 6 microsatellites may be a better tool for detection of bladder cancer.

Objective To evaluate the efficacy of retroperitoneoscopic adrenalectomy for pheochromocytoma.Methods From January 2000 to October 2006,a total of 16 patients(aged from 32 to 65 with a mean of 42 years) with pheochromocytoma received retroperitoneoscopic adrenalectomy in our hospital.Among the cases,6 had the tumor on the right side,and 10 on the left.The size of the tumors ranged from 2.5 to 4.6 cm in diameter(mean,3.1 cm).Results The mean preoperative preparation time in this series was 11 days(range,6 to 28).The retroperitoneoscopic adrenalectomy was completed in all but one of the patients,who were converted to open surgery because of extensive adhesion of the tumor to surrounding tissues and massive bleeding.The mean operation time was 110 minutes(90 to 170),and the mean blood loss was 135 ml(80 to 650).Three cases,who had normal blood pressure and thus received no noradrenalin immediately after the surgery,was given noradrenalin emergently 4,6,or 56 hours later owing to a sudden drop of systolic pressure(from 135 mm Hg to 80 mm Hg in 2,and from 140 mm Hg to 85 mm Hg in 1).Postoperative examination showed benign pheochromocytoma in 15 of the cases,and low-grade malignant pheochromocytoma with local invasion of the capsule in the patient who was converted to open surgery.The mean postoperative hospital stay was 12 days(9 to 20).The patients were followed up for 3 to 24 months(mean,13),during which only one received antihypertensive drugs;the others restored normal blood pressure spontaneously.No patient had abnormal levels of 24-hour urine noradrenalin,adrenalin,and catecholamine.Conclusions Retroperitoneoscopic surgery is an effective and minimally invasive treatment for patients with adrenal pheochromocytoma.The patients have a few complications and recover quickly after the operation.Preoperative preparation and postoperative treatment are important for the outcomes of the disease.