Objective:To investigate the generative or regenerative hair cell and studes of molecular and genetic in the inner ear,the long-term culture systems of utricular sensory epithelial cell of the rats were established.Method:Utricular sensory epithelial of postnatal day 1 rats was isolated by mechanical dissociation.The explants were digested by thermolysin,then transfrred to an aliquot containing trypsin and collagenase for incubation to harvest the pure utricular sensory epithelial cell. USEC wes cultured in Dulbecco modified Eagle medium(DMEM) and passaged.USEC of the 25 passages observed by inverted microscope and ultrastrctural examination with transmission electron microscope.Immunocytochemical method with cytokeratin 18 ,vimentin,Brn3.a and Calretinin; reverse transcription PCR with mRNA of AchRa9 and Myosin Ⅶa.The markers of hair cells were used identify the characteriztion of USEC of the 25 passages.Result:USEC have been cultured for more than 6 months and passaged 25 passages.USEC of the 25 passages showed a large,flat,polygonal epichlial morphotype with big,round neuclei.The cells grew into monolayer,cobblestone-likeappearance and showed Dome formation. The cells expressed cytokeratin 18 , not expressed vimentin,has rich microvilli and complex tight junction,which indicated the epithlial origination of USEC.Coexpressed of the hair cell characteristic markers Brn3.a,Calretinin and mRNA of AchRa9,Myosin Ⅶa was identified the culture cell that may represent rat progenitor hair cell.Conclusion:The long-term culture systems of utricular sensory epithelial cell of the rats were successfully established. The long-term culture USEC coexpressed the characteristic markers of the hair cell ,which identified the culture cell may represent progenitor cell of rats hair cell .It may provide valuable cell sources for in-depth investigation the mechanisms of hair cell generation or regeneration and studes of molecular and genetic in the inner ear.

BACKGROUND:It is reported that vitamin C can induce bone marrow mesenchymal stem cel s differentiating into osteoblasts, and promote bone repair and regeneration. However, vitamin C solution is unstable, so a carrier is necessary. OBJECTIVE:To observe the loading and control ed-release abilities of calcium phosphate used as the carrier ofvitamin C. METHODS:Calcium phosphate particles loaded with vitamin C were fabricated using chemical precipitation method, and the final concentration of vitamin C was 0, 0.1, 2 and 4 mmol/L, respectively. The drug-loaded capacity was detected. The release of vitamin from calcium phosphate nanoparticles in the simulate body fluid and ultrasonic environment was respectively evaluated. MC3T3-E1 cel s were co-cultured with calcium phosphate nanoparticles loaded with 2 mmol/L vitamin C, or calcium phosphate nanoparticles only. The cel proliferation was detected at 1, 3, 5 and 7 days of culture, and the alkaline phasphatase activity was detected at 1, 5, 10 and 15 days of culture. RESULTS AND CONCLUSION:The drug-loaded contents of calcium phosphate nanoparticles loading 0, 0.1, 2 and 4 mmol/L vitamin C were (59.9±5.4)%, (87.2±1.2)%and (28.4±26.3)%, respectively. Under normal environment, al samples could release vitamin C persistently, but the initial release speed of the particles carrying 0.1 and 2 mmol/L vitamin C was lower than that of particles carrying 4 mmd/L vitamin. Under ultrasonic environment, 2 mmol/L vitamin C-loaded calcium phosphate particles exhibited a quick release speed firstly that reached 5-15%, fol owed by a slow release speed. When ultrasonic powers kept at 75, 105 and 150 W, the release duration of vitamin C was 220, 340 and 260 minutes, respectively. MC3T3-E1 cel proliferation did not change after co-cultured with 2 mmol/L vitamin C-loaded calcium phosphate particles but the alkaline phosphatase activity was improved. These results suggest that calcium phosphate particles can be used as the carrier of vitamin C.

Objective To observe the expression of gamma-amino butyric acid A receptor(GABAA receptor)?3、?1、? subunits in rat flocculus following unilateral labyrinthectomy(UL).Methods 24 wistar rats were randomly divided into two groups.18 animals received unilateral labyrinthectomy while the others maintained labyrinthine well.After removing left labyrinthine,the change of GABAA receptor ?3、?1、? subunits was detected by immunohistochemistry.Results GABAA receptor ?1、? subunits in flocculus was induced on the operated side after unilateral labyrinthectomy.The most expression was on the 1st day flocculus of following UL.The expression is descending from the 3rd day to 7th day flocculus of following UL.The GABAA receptor ?3 subunit immunostaining in flocculus was weak and did not change in the process of vestibular compensation.Conclusion GABAA receptor ?1、? subunits were induced increase in the flocculus after unilateral labyrinthectomy.The alteration in the resting discharge of the central vestibular neurons may be caused by the increase of GABAA receptor ?1、? subunits in the flocculus.But the significance of the change of GABAA receptor ?1、? subunits in the vestibular compensation is still unknown.Our study suggests that GABAA receptor ?3 subunits may not participate in constructing GABAA receptor in the flocculus.

Objective To investigate the cell origination of progenitor cells of the mammalian regenerative hair cell.Methods Utricular sensory epithelial of postnatal day 1 wister rats was isolated by mechanical dissociation.The explants were digested by thermolysin,then were transfrred to an aliquot containing 0.125% trypsin and 0.125% collagenase for incubation to harvest the pure utricular sensory epithelial cell(USEC).USEC were cultured in Dulbecco Modified Eagle Medium(DMEM),and observed daily by inverted microscope.Immunocytochemical staining with cytokeratin 18,vimentin,Brn3a and Calretinin,ultrastrctural examination with transmission electron microscope,and reverse transcription PCR with mRNA of p27kipll,AchRa9 and Myosin Ⅶa.The markers of hair cells and supporting cells were used to identify the origination and characteriztion of USEC.Results USEC showed a large,flat,polygonal epithelial morphotype with big,round neuclei.USEC monolayer showed "cobblestone-like" appearance.Some cells showed "Dome" formation,probably due to fluid collection underneath the cell monolayer.USEC expressed cytokeratin 18 but did not express vimentin,and had rich microvilli and complex tight junction,which indicated the epithelial origination of USEC.USEC expressed Brn 3a,calnevimin and mRNA of p27kipl,AchRa9,myosin Ⅶ.Conclusion USEC coexpressed the characteristic markers of the hair cell and supporting cells.This indicates that the culture cells may represent progenitor cell of hair cell and the progenitor cell may come from supporting cells.

Objective To investigate the possible association of mtDNA deletion in spiral ganglion of cochlear and cochlear nucleus of brain in young and elder rat with presbyacusis.Methods The mtDNA 4834 deletion in spiral ganglion of cochlear and cochlear nucleus of brain in rat was analyed by PCR.Results The deletion rate of mtDNA 4834 in rat with presbyacusis was 66.7% in spiral ganglion, 100% in cochlear nucleus of brain.While no deletion was detected in young rat.Conclusion Deletion in mtDNA of the spiral ganglion of cochlear and cochlear nucleus of brain was found in rat with presbyacusis, and it may play an important role in the pathogens of presbyacusis.

ObjectiveTo investigate the cause, treatment and prevention of splenic injury in radical nephrectomy for left renal cell carcinoma.MethodsThe clinical data of 27 cases of splenic injury in 458 cases of radical nephrectomy for renal cell carcinoma were retrospectively analyzed.ResultsThe injured spleen was saved in 21 cases (18 gradeⅠ injury,3 grade Ⅱ).The injured spleen was sewed up and pressed with hemostatic ganze in 7,pressed with medical sponge laid with spurting argon on biological glue in 11 and the same was carried out in 3 grade Ⅱ injury.Spleenectomy was conducted in the other 6 cases (4 grade Ⅱ injury,2 grade Ⅲ).The patients have been followed up for 6 months to 5 years.The outcome has been good in all except 1 patient died of tumor recurrence 1 month after operation.No late bleeding or adverse side effects has been noted.Conclusionssplenic injury may occur in radical nephrectomy for renal cell carcinoma especially the tumor is lasge or adhered to spleen in the upper past of left kidney.The occurrence of splenic injury is also related to the tumor stage.Attention should be called to the mentioned surgical complication.

Objective To probe the effect of Cpd danshen tablet combined with vitamin E in liver fibrosis by chronic hepatitis B.Methods 44 patients with CHB-related live fibrosis were allocated randomly into control group and Cpd danshen tablet combined with vitamin E treatment group for 6 months' treatment.Serum ALT,AST level and fibrosis markers including hyaluronic acid (HA),Ⅳ collagen(Ⅳ-C),amino terminal propeptide of type Ⅲ procollagen (PⅢP),and laminin(LN) and serum MMP1,2,TIMP1,2 level were determined before and after 6 months.Liver biopsies before and after 6 months were assessed.Results ALT,AST level in the two groups were decreased after 6 months' treatment(.P.

Objective To investigate perinatal outcomes of twin pregnancy.Methods Perinatal outcomes of 658 pregnancies with twins hospitalized and born at Beijing Obstetrics and Gynecology Hospital during 2002 to 2005 were retrospective analyzed to compare the influence of type of twin,delivery mode, gestational week,birth time interval between twins,as well as specific complications of twin pregnancies on their outcomes.Results Pefinatal fatality of monzygous twins was 53.69 per thousand(16/298),higher than that of dizygous twins(27.11 per thousand,9/332),with U=2.35,P0.05.Perinatal fatality was higher in twins with different development (birth weight).Perinatal fatality in dizygous twins(12.5 percent,2/16)was lower than that in monozygous ones(14.3 percent,6/42),and that in twins with different development(birth weight)(0.6 percent, 2/316)was lower than that with same development(1.2 percent,3/256),with ?~2=16.944,P

Objective To retrospectively investigate the impact of postoperative radiotherapy ( RT) on the relationship between molecular subtype and survival in patients with breast cancer ( BC ) . Methods A total of 716 women who were admitted to our hospital in 2008 and newly received unilateral mastectomy were divided into Luminal A ( LA ) , Luminal B?HER?2?negative ( LB1 ) , Luminal B?HER?2?positive ( LB2) , HER?2 overexpression ( HER?2+) , triple?negative ( TN) , and unassigned subtypes according to the 2011 St. Gallen Consensus. The Cox model was used to analyze the differences in overall survival ( OS) and disease?free survival ( DFS ) rates between subtypes in all patients, RT group, or non?RT group. The Kaplan?Meier method was used to calculate OS and DFS rates. The Cox model was used to perform the factor analysis. Results In all patients, the median follow?up time was 71?4 months;the overall mortality rate was 10?5%;the incidence of treatment failure ( death+relapse+metastasis) was 14?9%;217 patients ( 30?3%) received RT. The multivariate analysis showed that there was no significant difference in OS between subtypes in any group ( all P>0?05 ) . In all patients, patients with LB1 subtype or unassigned subtype had significantly poorer DFS rates than those with LA subtype ( HR= 1?881, P= 0?035;HR= 1?907, P=0?049) . In the non?RT group, patients with LB1 subtype had significantly poorer DFS rates than those with LA subtype (HR=3?324, P=0?01). In the RT group, there was no significant difference in DFS rate between subtypes ( all P>0?05) . The two?dimensional cross analyses of RT and subtype demonstrated that patients with LB1 subtype in the non?RT group had lower OS and DFS rates than patients with LA subtype in the RT group ( P=0?09,0?06) . Conclusions Patients with LB1 subtype have lower OS and DFS rates than patients with LA subtype, especially in the non?RT patients. RT has no impact on the relationship between subtype and prognosis.

Taxus canadensis distributed mainly in North America, such as northern Minnesota, Newfoundland, south to Wisconsin and Pennsylvania. Its composition has been shown to be very different from other species, and in recent years, some new skeletons also have been found in Canada yew. Through analysis of the taxanes content on various Taxus plants containing taxanes, the results showed a higher content of taxol in T. canadensis. Based on the current research on T. canadensis (from the research results of the author in recent years, as well as from studies of scholars in the field), the paper outlined the research progress in recent years on the chemical constituents of taxane of T. canadensis and the spectral characteristics of various types of compounds. Besides, this paper analyzed the present research about solutions for the taxol drug source crisis.
Animals ; Bridged-Ring Compounds ; chemical synthesis ; chemistry ; pharmacology ; Humans ; Molecular Structure ; Plant Extracts ; chemical synthesis ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry ; Taxoids ; chemical synthesis ; chemistry ; pharmacology ; Taxus ; chemistry