Objective: To study the transdermal releasing rule of the preparation of Jiufen Spray.Methods: Improved Franz diffuser was applied to transdermal experiment with TLC scanning method.Results: Q T (quantum time) equations of Jiufen Spray are strychnine:Q=220.941t- 486.006 , brucine:Q=208.146t-454.629, ephedrine:Q=177.691t-247.826. The study on releasing rule suggests the accumulative amount of transdermal drug increases with time, but the releasing speed is roughly stable, the total releasing ratio of 12 hours is about 50%. Conclusion: Jiufen spray could surmise to maintain relatively stable plasma drug concentration.

Objective To compare the clinical application effect of bi -level positive airway pressure ( BiPAP) and nasal continuous positive airway pressure ( CPAP) in the treatment of preterm infants with neonatal respiratory distress syndrome ( NRDS) .Methods 64 neonates with NRDS were divided into the BiPAP group ( n=35) and NCPAP group (n=29) according to the randomized controlled study method.The blood gas index before and after 2,12 hours of treatment,the total time for ventilation,length of stay,and the incidence rate of abdominal disten-sion,gastroesophageal reflux,bronchopulmonary dysplasia,intraventricular hemorrhage,ventilator-associated pneumo-nia in the two groups were compared.Results The levels of pH and PaO2 in the BiPAP group were higher than those in the NCPAP group in noninvasive respiratory support for 2,12h(t=2.391,2.556,2403,2.355,P=0.020,0.013, 0.019,0.022).The incidence rates of hypoxemia,apnea,re-intubation mechanical ventilation rate after 12h in the BiPAP group were lower than those in the NCPAP group(t=5.049,4.988,4.215,P=0.025,0.026,0.040).The length of stay and mechanical ventilation time in the BiPAP group were shorter than those in the NCPAP group( t=-2.096,-2.669,P=0.041,0.010).Conclusion Compared with NCPAP,early application of BiPAP can obviously reduce the invasive respiratory support rate of children with NRDS with intubation.It is worthy of promotion.

Objective: To screen the receiving solution used to study the transdermal absorption of strychnine in Jiufen spray. Methods: With recovery,Q-T equation and transdermal speed constant as the parameters, the improved Franz-cell and the skin of SD rats were used to screen the receiving solution from saline, saline-alcohol (7∶3,v∶v), pH phosphate buffer solution-alcohol (88∶2,v∶v), pH 7.4 phosphate buffer containing 25% alcohol. Results: The comparison of the inter-day and intra-day recoveries,Q-T equation and transdermal speed constant showed that saline-alcohol(7∶3) was the best for the study. Conclusion: It is indicated that saline-alcohol(7∶3) was the best receiving solution for the transdermal absorption study of Jiufen spray.

Objective To observe the effect of volume resuscitation on the early grafted kidney function in opration.Methods 909 cases were divided into five groups according to the volume resuscitation in opration:group A:

[Objective]To investigate the effect of salvianoli acid B and hyperoxic solution on experimental ischemia reperfusion injury of extremities.[Method]Twenty-four healthy New Zealand rabbits were divided into 4 groups randomly.Equivalent 0.9%Nacl(group A),hyperoxic solution(group B),salvianoli acid B(group C),salvianoli acid B and hyperoxic solution(group D) were injected through ear edge vein before ischemia.The femoral artery and vein were blocked and the model of ischemia reperfusion injury of extremities was set up.Before ischemia and after 4 hour's reperfusion,the blood samples were taken to measure the levels of blood malondialdehyde(MDA) and superoxide dismutase(SOD).The gastrocnemius muscles were observed by pathologic test.[Result]Serum enzymatic activity of MDA improved obviously.Salvianoli acid B and hyperoxic solution could prevent the improvement of it(P

Objective To study the effects of intravenous immunoglobulin (IVIG) on neonatal lymphocyte immune function. Methods Cord blood mononuclear cells (CBMCs) and CD3+ T lymphocytes were isolated from 8 neonates and were cultured with IVIG and/or/without phytohemagglutinin (PHA)based on which divided into five groups, i. e. (l)control group, (2)PHA activation group, (3) IVIG pre-inhibition group, (4) PHA pre-activation group and (5) PHA+IVIG group. The production of IFN-? and IL-4 of CBMCs and cord blood CD3+ T lymphocytes was measured with ELISA and the expression of IL-2 and IL-4 mRNA was measured with RT-PCR. The results were compared with peripheral blood mononuclear cells(PBMCs) of 8 adults. Results The production of IFN-y by CBMCs and cord blood CD3+ T lymphocytes induced by PHA decreased after IVIG was given, while more remarkable decrease in the CBMCs was shown without any association with IVIG was given or not before or after PHA was used [Group 2; (313. 34?108. 00)pg/ml, Group 3: (17. 68?17. 98) pg/ml, group 4 : (170. 72?38. 25) pg/ml, group 5: (11. 10?8. 92)pg/ml]. For the CD3+ T lymphocytes, the decrease of IFN-? production was only observed in group 3, 4 and group 5 [(35. 30 ?12. 21)pg/ml, (8. 61?6. 21) pg/ml and (8. 54?1. 57)pg/ml]. The degree of decrease of IFN-? production by CBMCs was much lower [group 2: (1121. 11 ? 344. 58)pg/ml, group 3: (29. 08 ? 11. 20)pg/ml, group 4: (339. 63?47. 43) pg/ml, group 5: (26.40?21. 97)pg/ml]. The secretion for IL-4 did not changed when stimulated by PHA alone, but IL-2 and IL-4 mRNAs expression can be detected in CBMCs after repeated stimulation by PMA, PHA and rIL-2 which decreased after IVIG was given. Conclusions IVIG can suppress the production of IFN-? in cord blood T lymphocyte directly or mediated by other immune cells or molecules. IVIG can also decreased the expression of IL-2 mRNA and IL-4 mRNA in CBMCs. The effect of IVIG on the proliferation of cord blood lymphocytes and production of immunoglobulin in B lymphocytes might be associated with these inhibition.

A case of benign follicular neoplasm-trichogerminoma-is reported.A 48-year-old man presented with a 10-year history of asymptomatic subcutaneous nodule in the chest.Histological examination revealed a well-circumscribed lesion composed of variously sized lobuli and cysts in the deep dermis and separated from the surrounding tissue by a fibrous capsule.Most lobuli consisted of concentrically arranged clear cells in the central area and basophilic cells in a palisade arrangement in the peripheral area.The tumor cells displayed a multi-directional differentiation toward hair bulb,inner root sheath,outer root sheath and infundibulum of hair follicles.Immunohistochemically,the tumor cells expressed AE1/AE3,CK5/6 and CK17,but were negative for CK20 or CK7.There was a sharp contrast in immunohistochemical findings between the central clear cells and peripheral basophilic cells.Based on the histological and immunohistochemical features,a diagnosis of trichogerminoma was made.

BACKGROUND: The transplantation of allogeneic cartilage has local immunological rejection, and it is necessary to further reduce the rejection to promote its application in clinic, thus it is significant to perform a series of experiments to induce local immune privilege.OBJECTIVE: To observe the in vivo growth of tissue engineered allogeneic cartilage reconstructed by chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL.DESIGN: A randomized controlled observation.SETTING: Shanghai Jiao Tong University.MATERIALS: Thirty-six allogeneic New Zealand rabbits as recipients and 45 1-week-old chinchillas as donors, either sex,were purchased by the experimental animal center of Chinese Academy of Sciences. Amphotropic recombinant retrovirus coated cell line PT67 was purchased from Clontech Company; Dulbecco's modified eagle medium (DMEM), fetal bovine serum (FBS), G418 and Polybrene were bought from GIBCO BRL.METHODS: The experiment was carried out in original Shanghai Second Medical University from January 2000 to July 2005. The New Zealand rabbits were randomly divided into three groups: FasL-transfected group (n =12), untransfected group (n =12) and blank control group (n =12). The rabbit allogeneic cartilages were constructed by the compound of pLNCX2-FasL transfected chondrocytes and tissue engineered material of pluronic F-127. ① Gross observation and mass changes of the grafts: Corresponding materials were infused subcutaneously, the grafts were removed at 1, 2 and 3 months after transplantation for gross observation and the mass changes. ② Staining observation: The grafts were removed at 1, 2 and 3 months after transplantation, then prepared into sections, and observed by hematoxylin and eosin (HE), Safranin'O and Masson's trichrome stainings. ③ Antibody detection: Blood samples (1 mL) were collected at 1 and 2 months after transplantation, the chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, and separated by electrophoresis in agarose medium, then acted with serum of recipient to observe whether corresponding antibody generated. ④ Complement dependent cytotoxicity (CDC) test: The chondrocytes of chinchillas were prepared into cell suspension (2×109/L), and then seeded into 96-well plate, attached grew for 24 hours, then recipient serum was added for the CDC test, and the percentage of apoptotic cells was counted under microscope.MAIN OUTCOME MEASURES: ① Gross observation and mass changes of the grafts;② Histological changes; ③ Results of the antibody detection; ④ Percentage of apoptotic cells.RESULTS: All the 81 rabbits were involved in the analysis of results. ① Gross observation and mass changes of the grafts: Two weeks after inoculation, there were obvious nod formations at the inoculated sites, but no nod formed in the blank control group. The new cartilage tissues became smaller gradually and completely disappeared at 4 months in the untransfected group, whereas those in the FasL-transfected group became smaller, but still existed after 7 months. The masses of grafts in the FasL-transfected group were higher than those in the untransfected group (P ＜ 0.05). ②Histological observation: Plenty of lymphocytic infiltrations around cartilage tissue could be observed in the untransfected group, and obviously decreased in the FasL-transfected group. No lymphocyte was observed inside the chondrocytes.Masson's trichrome staining was performed, and it was observed under light microscope that the small white parts in the middle were immature chondrocytes, and there were green collagen around most of the mature chondrocytes. Safranin O staining showed strong positive reaction, suggested that there were rich glycosaminoglycan in matrix. ③ Antibody detection: The chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, then acted with serum of recipient, and the results showed that no corresponding antibody generated. ④ Percentage of apoptotic cells: The percentages of serum CDC apoptotic cells in the FasL can ransfected group, untransfected group and blank control group were 5%, 6% and 1%, which were all negative.CONCLUSION: Rabbit allogeneic chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL can reconstruct tissue engineered cartilage, and can postpone the degeneration by 3 months.

BACKGROUND: At present,enhanced green fluorescent protein(EGFP)is proved to be the best labeled molecule,with unique advantages,such as high fluorescence specificity,easy to be detected,and so on.Recombined retroviral vector EGFP-pLNCX2,which can stably express EGFP,can be construct using gene-engineering technique.Transfecting allogenic chondrocytes is indeed very useful to investigate the target gene expression and process of constructing tissue engineered cartilage in vivo.OBJECTIVE:To construct recombined retroviral vector EGFP-pLNCX2 which can stably express EGFP,and investigate optimal conditions for retrovirus transfection of chondrocytes.DESIGN:Randomized controlled observation.SETTING:Shanghai Jiao Tong University.MATERIALS:Thirty New Zealand rabbits of either gender,1 week of age,were purchased from Experimental Animal Center of Chinese Academy of Sciences.Amphotropic retrovirus package cell line PT67 pLNCX2 and pEGEP-C1 were purchased from Clontech Company;NIH 3T3 cell line was purchased from ATCC Company;DH5a Bacterium coli was preserved by the laboratory of Shanghai Jiao Tong University;Retroviral vector pLNCX2 was purchased from Clontech Company;pEGFP-C1 plasmid with EGFP were donated by professor Cong Xiao-qian from Chinese Academy of Sciences.METHODS:This experiment was carried out in the Shanghai Jiao Tong University in August 2005.Chondrocytes of New Zealand rabbits were isolated and cultured.The recombinant retroviral vector EGFP-pLNCX2,which can stably express EGFP,was constructed to transfect cultured chondrocytes from New Zealand rabbits by using gene engineering technique.Transfection results were observed under fluorescence microscope.Altogether 6×105 chondrocytes incubated in 10 cm-diameter flat plate were used to transfect retrovirus-EGFP immediately and at 12,24 and 48 hours after inoculation.One week later,EGFP expression efficiency was measured with flow cytometer,and best occasion for retrovirus transfecting primary chondrocytes.Enzyme-digested chondrocytes were inoculated for 24 hours to transfect retrovirus.After 250 mg/L G418 was added,chondrocytes were screened on the 2nd,3rd,4th,5th and 6th days separately.After phosphate buffer solution(PBS)washings,the transfection efficiency of chondrocytes was detected by flow cytometer,and the best occasion for G418 screening was observed.MAIN OUTCOME MEASURES:①EGFP-pLNCX2 transfection efficiency.②The best occasions for retrovirus transfecting primary chondrocytes and G418 screening.RESULTS: ①Retroviral vector EGFP-pLNCX2 transfected primary chondrocytes of rabbits,and high expression of transfected chondrocytes could be obtained through preliminary screening of G418.After being screened and expressing EGFP,chondrocytes kept normal morphology,with pseudopod adhering to the wall and matrix secreting vigorously.②The best occasion for retrovirus transfecting primary chondrocytes was at 24 hours after cell inoculation.The transfection efficiency determined with flow cytometer was 19.14% on the 5th day.The best occasion for G418 screening was on the 5th day after culture.Transfection efficiency of G418 screening was 55.75% on the 7th day.CONCLUSION:Recombinant retroviral vector EGFP-pLNCX2 can effectively transfect chondrocytes.The best occasion for retrovirus transfecting primary chondrocytes is at 24 hours after inoculation, and the best occasion for G418 screening is on the 5th day after culture.

Objective To estimate the diagnostic value of interstitial infiltration pattern for early morphea.Methods Twenty-five cases of early morphea pathologically characterized by interstitial infiltration of inflammatory cells were collected from 2010 to 2012.The clinicopathological features of these cases were retrospectively analyzed.Results The average clinical course was 7.5 months.The primary manifestation was edematous dark erythematous plaques,and interstitial or mixed infiltrate of inflammatory cells was the characteristic histopathological presentation.After anti-inflammatory treatment,lesions markedly improved or disappeared in 70％ of these patients.Conclusions Interstitial infiltration of inflammatory cells is a rare histologic pattern in early morphea.To learn and recognize this pattern may be beneficial to the diagnosis and treatment of early morphea.