Mitral effective regurgitant orifice area was measured by using color Doppler flow convergenee method,and its value on quantilarian of mitral insufficiency Was evaluated.The results showed that the parameter was correlated well with regurgitant volume,regurgitant fraction,color Doppler jet beginning width and jet area(r=0.89、0.84、0.82 and 0.67 respectively,P<0.001～0,0001).The accurate rate in differentiating mild and moderate from severe mitral regurgitation was 95.7% with the effective regurgitant orifice area≥30mm2.The effective regurgitant orifie area is a good parameter in quantification of naitral insufficiency.

The value of flow convergence and jet parameters in quantification of mitral regurgitation was estimated.The resultS showed that there were excellent correlations between mitral regurgitant flow convergence volume and mitral regurgitant volume,regurgitant fraction(r=0.87～0.93,P<0.0001).The correlations between jet parameters and regurgtant volume,regurgitant fraction were different,jet beginning width:r=0.88 and 0.81(P<0.0001),jet area : r=0.72 and O.50 and 0.61(P<0.05)and the ratio of jet area to left atrial area:r=0.27 and 0.29(P>0.05).The accurate rates of regurgitant flow convergence volume and jet beginning width in differentiating the grades of mitral regurgitation were 91.3～95.7%and 82.6～95.7%respectively.Mitrai regurgitant flow convergence volume is an accurate and reliable parameter in the quantification of mitral regurgitation.Jet parameters are of relative value in semiquantitation of mitral regurgitation,especially the jet beginning width.

Flow convergence is a new method of color Doppler flow quantification.The effect of instrumental settings on flow convergence was investigated in an in-vitro model.The result showed that the flow convergence volume increased from 1.67±1.01 l/min to 2.20±1.35～2.30±1.43 l/min(P< 0.0001)when aliasing limit(same concept as Nyquist frequency)was changed from 23 cm/s to 12cm/s. There was no significant effect on flow convergence volume with the change of other instrumental settings,including Color gain,frame frequency,paket size,transmit power,transducer frequency,etc. Color Doppler flow convergence is influenced by aliasing limit.mainly because that the isovelocity surfaces in acceleration region developed by different aliasing limits are not always in the shape of hemisphere.The result suggested that besides the selection of aliasing limit fits to a hemispherical isovelocity surface,flow convergence is rarely affected by the other instrumental settings,and therefore is a relatively stable parameter of flow quantification.

Objective To isolate,culture and identify epidermal stem cells(ESCs).The gene of enhanced green fluorescent protein(eGFP) was introduced via recombinant adeno-associated virus(rAAV) infection into the epidermal stem cells in vitro and the transfection efficiency under various tite of culture was determined. Methods 1.Getting fetal Wistar rats'dissociated single epidermal cells.2.Making laminin and CollagenⅣ the substitution of basal memrane,ESCs were isolated by adhering to murine laminin and CollagenⅣ.3.Epidermal stem cells were cultured in twenty-four-well plate,and cells number was controled 5?10~4 or so.After epidermal stem cells adhered to plate,diluted rAAV_2/eGFP virus fluid with serum-free medium.The diluted rAAV_2/eGFP virus fluid was added to the twenty-four-well plate according to the different MOI(viru gene/cells).The number of green fluorescence cells were counted under fluorescence microscope.The transfection efficiency was determined. Results 1.ESCs had better adhesive ability to laminin and higher colony formation efficiency(CFE) than that of keratinocytes.2.ESCs wer strongly positive with immunocytochemical staining of integrin ?1 and keration 19(K19).3.After rAAV_2/eGFP genetic transfection of ESCs,the positive cloning expressed highly eGFP gene along with time.Conclusion 1.These results suggest that rats'epidermal stem cells could been successfully isolated and enriched in vitro by means of rapid adherence to laminin and CollagenⅣ.After rAAV_2/eGFP genetic transfection of epidermal stem cells,the positive cloning expressed highly eGFP gene stably along with time.

To explore new teaching method of functional experiment,we opened the functional laboratory to encourage students' creativity and train their practical ability and develop their innovative ability.The experiment teaching innovation was attempted and discussed the functional speciality.

Objective To investigate the CT imaging features of pneumonic type Bronchio-loalveolar carcinoma(BAC) and the value of CT-guided percutaneous puncture biopsy in pneumonic type BAC.Methods Twenty-five cases of suspected pneumonic type BAC were biopsied with CT-guided percutaneous puncture.GE 64-slices plain and post contrast CT scans were performed in all patients.Two radiologists finished imaging determination.Site of puncture was selected in suspicious areas of the tumor.The negative pressure aspiration needle were used to be biopsy in the ideal location after vertically into the needle and again performed CT scanning were performed to observe the complications such as hemorrhage,pneumothorax.Specimens were fixed by formalin.Results Twenty-five samples were successfully got from 25 patients.Pathology data showed that 14 cases were pneumonia with BAC,6 cases with pneumonia,2 cases with offungus infection,and 3 cases with caseous pneumonia.Of 14 cases (71.4％) with pneumonia type BAC,CT images of 10 cases showed air bronchogram of lung tissue,5 cases(35.7％) with alveolar gas cavity or cavity of lung tissue,5 cases(35.7％) with ground glass opacity and multiple nodules.Enhanced CT scanning the area of consolidation showed that 11 cases(78.6％) were with mixed low density area angiography and the mean peak time was 90 s,9 cases (64.2％) were with the time density curve of speed up and slow down type.Conclusion The method of CT guided lung biopsy combined with typical imaging findings can enhance early diagnosis rate of pneumonia type BAC.Meanwhile CT feature of pneumonia type BAC shows honeycomb air cavity,void,angiographic sign,dead branches syndrome,multiple peripheral acinar nodules and ground glass density.Enhanced scan area of consolidation shows visible angiogran sign,slow drop type of time density curve of a fast rise.

This investigation was made on 30 adult female rats weighing approximately 150～200g. during estrus as judged from vaginal smear examination. Ovaries which investigated with electron microscope, were fixed in 3％ glutaraldehyde. Other ovaries were embeded in paraffin and examined under light microscope.Theca folliculi of large antral follicles were examined with electron microscope. Three kinds of theca cells were distinguished. They were secretory cell, fibroblast and smooth muscle cell. The secretory cell showed the characterastics of the steroidproducing cell, having smooth endoplasmic reticulum, mitochondria and lipid droplets. The fibroblast contained rough endoplasmic reticulum, free ribosomes and mitochondria in the cytoplasm. The smooth muscle cell was located in the outer layer of theca folliculi, and its structural features were typical according to the criteria of Somlyo and Somlyo: (1) pinocytotic vesicles of the cytoplasmic membranes; (2) myofilaments in the cytoplasm; (3) dense bodies intimately related to the myofilaments; and (4) location of mitochondria and endoplasmic reticulum at the nuclear poles.The ultrastructure of smooth muscle cell, its differentiation and other problems were discussed.

The report of ultrastructure of dog ovarian follicle wall has not been seen yet. This investigation was made on 5 adult dogs in the estrus stage of the cycle and examined with electron microscope. There are granular cells, fibroblasts and secretory cells in the ovarian follicle wall, The typical smooth muscle cell is absent.

Objective To identify the waves of abnormal myocardial Doppler tissue imaging (DTI) spectra and their alterations.Methods In 13 open chest dogs left ventricular (LV) wall motion velocity patterns at the mid wall portion of the middle of the LV posterior walls in the short axis direction were recorded with DTI before and after arteries infarct.Simultaneously the LV pressure (P) curves,their first derivative (dP/dt) curves,electrocardiography (ECG) and phonocardiogram (PCG) were recorded.Compared with ECG,PCG,P curves,and dP/dt curves the waves of abnormal myocardial DTI spectra and their alterations could be identified.Nineteen patients with myocardial infarction and six cardiac allograft recipients entered the clinical study.The motions of LV walls and mitral annuli were studied with DTI spectra and tissue velocity imaging (TVI) curves.Basing on ECG the velocity waves of DTI spectra and TVI curves were identified.Results In dogs the acutely ischemic walls motioned negatively(IVC 2) while the normal LV wall motions were mainly positive (IVC 1) during isovolumic contraction and these negative motions could prolong into injecting phase.Before and after coronary arteries occlusion,acute ischemia resulted in significant reduction of peak IVC 1S,and A [( 16.60 ? 4.11 )cm/s vs ( 4.60 ? 5.38 )cm/s,( 11.30 ? 1.58 )cm/s vs ( 7.16 ? 1.80 )cm/s,( 11.08 ? 3.35 )cm/s vs ( 6.71 ? 4.81 )cm/s,respectively,P