Spleen cells of BALB/C mice immunized with medical urokinase were fused with FO and NS-1 myeloma cells three times, and six hybridoma cell lines(I-UB5, I-UD2, 3-UA2, 3-UD12,3-UG2 and 3-UH7)secreting monoclonal antibodies against urokinase were obtained after screening.Of them, I-UB5 and I-UD2 v/ere cloned, frozen, stored, resuscitated and analysed chromosomally.It was found that I-UB5 and I-UD2 were clones obtained by fussing spleen cells of BALB/C mice with FO myeloma cells.Their subclasses are IgG1 and IgG2a respectively determined by immunodifussion and enzyme-linked immunosorbent assay(ELISA).The titers dctc-termined by ELISA were in the range of 10-2-10-3 in culture supernatants, and 10-6-10-7 in ascitic fluid.The I-UB5 and I-UD2 were proved to be monoclonal antibodies against high molecular weight subunit of urokinase, since they did not react with the purified low molecular weight subunit of, but reacted with mixed subunit of urokinase

Objective To generate an human interleukin-17 receptor-like molecule (IL-17RLM) recombinant plasmid with 6?myc tag and detect its specific expression in eukaryotic cells. Methods Design two specific primers(including the enzyme sites of EcoRⅠand XhoⅠ), reextract hIL-17RLM-L DNA fragment after PCR and insert it into the 6?myc tagged pcDNA3.0 vector, then detect its expression by Western blot after transfecting COS7 cells. Results The 6?myc tagged recombinant plasmid pcDNA3.0- 6?myc /hIL-17RLM-L was generated successfully and its expression can be detected by Western blot in eukaryotic cells. Conclusion The eukaryotic expressing plasmid pcDNA3.0-6?myc /hIL-17RLM-L was generated successfully and its specific expression was realized, which may provide the basis for further research of its biological function.

Objective To explore the changes of matrix metalloproteinase-9 and blood brain barrier in cardiopulmonary resuscitation rats and effects of MMP-9 inhibitor on them.Method One hundred and twenty Sprague-Dawley(SD) rats were randomly divided into 3 groups:the sham-operated group,the resuscitation with treatment group and the resuseimfion without treatment group as control.The experiment was made in the animal experiment center of Sun Yat-sen University in Gtlangzhou.The rat eardiopulmonary resuscitation model was made by clipping trachea until asphyxia,and the restoration of spontaneous circulation(ROSC)Was defined by restoration of superventricular rhythm and mean artery pressure (MAP)≥60 mmHg for more than 5 min utes.The rats of sham-operated group were anesahetized only and endotracheal intubation WaS performed.In the resuscitation with treaUnent group ss-3cr(25,ng/ks body weight)Was given intraperitoneally after ROSC.The rats were sacrificed and samples of the brain tissue were taken inmaediately and 3 h,9 h,24 h and 48 h later.After that,the expression of MMP-9 and MMP-9 mRNA in brain tissue were detected.Water oontent and Evans blue in brain tissue Were observed.The uhmmicrostructure of brain tissue was observed under electron microscope.Analysis ofvariance wilE, done with Spssll.0 software.Results 11le expressions of MMP.9 and MMP-9m RNA ofbraintissueiUthe shanloperated group didn't show significant changees in all specimens taken at different intervals and neither the water content and tvans blue did.The Pvalue were 1.0000,0.6831,0.7124 and 0.99r75,respectively.There was no u1.tramicrostruclure change in the sham-operated group.The expressions of MMP_9 and MMP-9 mRNA in the resuscitation control group obviously increased after eardiopulmonary resuscitation,80 did the water content and Evans blue content.Compared with sham-operated group,the P value were 0.0264,0.0163,0.0000 and 0.0412,respee.tively.111e elge of ultmmicrostmeture in the resuscitation control group at different intervals were obvious.The changes of obove biomarkers in the resuscitation treatment group Was siroilar to but less in magnitude than those in the resuscitation control group.The P valHe were 0.0392,0.0373,0.O004 and 0.0180,respectively.Conclusions The expressions of MMP-9 and MMP.9 mRNA obviously increases in the cerebral ischemia model of rats with CPR,and reaches peak at 24 h.Water content and Evans blue content in brain risque obviously increases in the cerebral ischemia model of rats with CPR.BBB iS destroyed.and the peak time iS at 24 h.The injury of ultrami.crostructure of brain tissue under electron microscope iS obvious,and the peak time is at 24 h.The SB-3CT.specif-iC inhibitor of MMP-9 could decrease the expression of MMP-9 and decrease cerebral edema in the cerebral is.chemia modeJ of rats with CPR,and the protection from cerebral isehemia/reperfusion injury after CPR is obvious.

Objective To investigate re-innervation in the neovaginal mucosa of patients underwent sigmoid colon vaginoplasty in treatment of Mayer-Rokitansky-Kistner-Hauser Syndrome (MRKHS).Methods Biopsies in the upper third of the posterior neovagina were taken in 20 patients treated by sigmoid colon vaginoplasty at 1, 2 and 3 years after surgery, respectively. Protein gene product 9. 5 ( PGP 9. 5 ),vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY) were detected by immunohistochemical method and compared with those in intact sigmoid colon mucosa. Results ( 1 ) Density of nerve fiber:abundant distribution of PGP 9. 5 nerve fibers were observed in the mucosal muscle layer, submucosa, and smooth muscle layer of the neovagina. The nerve fibers of VIP and NPY immunoreactivity were mainly distributed around blood vessels and in the smooth muscles. In the neovagina, the density of nerve fibers of PGP 9. 5 of 17 ± 6 were much more than VIP of 2. 9 ± 1.0 and NPY of 2. 5 ± 0. 8 significantly ( P ＜ 0. 05 ).( 2 ) Expression of PGP 9. 5 in neovagina: at 1 year after surgery, PGP 9. 5 positive expression of 14 ± 4 was significantly lower in the neovagina than 28 ± 7 in the intact sigmoid colon( P ＜ 0. 05 ). However, after 2 to 3 years, its expression displayed an upgrade tendency in the neovagina and was significantly higher at the 3 year postoperatively than that at the 1 years postoperatively ( 22 ± 7 vs. 14 ± 4, P ＜ 0. 05 ). The changes were much more obvious in submucosa. (3) The expression of VIP and NPY in neovagina: at 1 year after surgery, VIP and NPY positive nerve fibers were also decreased in the neovagina when compared with those in the intact sigmoid colon ( 2. 3 ± 0. 7 vs. 5.3 ± 1.4, P ＜ 0. 05; 2.5 ± 1. 1 vs. 5.5 ± 1.1, P ＜ 0. 05 ) . At 2 to 3 years after surgery, the positive VIP fiber showed initially decreased and subsequently increased tendency. The density of VIP of 3.7 ± 0. 7 in the neovagina at 3 years postoperatively was higher than 2. 3 ±0. 7 at 1 years postoperatively (P ＜ 0. 05 ). No significant up-regulation was observed in NPY-positive expression in the neovagina within 3 years after operation. Conclusions Distribution of sensory PGP 9. 5,VIP and NPY immunoreactive nerve fibers was similar to the pattern observed within the intact sigmoid colon wall. The number of nerve fibers in the neovagina decreased after surgery and then increased subsequently within 3 years after surgery.

quamous metaplasia occurred, the expression of occludin and ZO-1 in cells were similar to those in control group, which might play a role in the defense function of neovaginal mucosa.

Objective To explore the effect of online health education by family physician using the wearable devices, and provide the basis in theory and practice to improve the level of primary health care in community. Methods A total of 132 community residents (78 for male and 36 for female, all of them are WeChat user) with hypertension in one residential area who had regularly participated in the activity of volunteer blood pressure measurement organized by community health service center in wujiaochang street from November 2015 to February were chosen as the participants. The residents who wore the wearable device given by the community hospital uniformly were set as the study group (60 people), while the remaining 72 residents were set as the control group. Both groups received the daily online health education provided by the family physicians through their smartphone. We compared the diversity of health cognition, controlling behavior and controlling results to blood pressure of both groups before and after the studyrelatively, then we comprehensively analyzed the effect of use of wearable devices in family physicians ' online health education. SPSS 18.0 was used for data analysis. The categorical data of two groups were compared with chi-square test and the comparison of continuous data was performed with t test. Results Totally 114 community residents were qualified in the study, and the initial backgrounds of study group and control group had no significant difference. After receiving the online health education information for three months, both groups achieved improvement on the cognition and related self-control behavior of hypertension except for several items, and the study group did better than control group in most items. The physical examination showed that the systolic blood pressure of the study group after the intervention [(131.46 ± 12.89) mmHg] (1 mmHg=0.133 kPa) was lower than before [(137.24 ± 12.27) mmHg] and the difference was significant (t=9.2, P<0.01); the diastolic blood pressure of the study group after the intervention [(78.29±8.91) mmHg] was lower than that of before intervention [(80.75±10.25) mmHg] and the difference was significant (t=2.3, P<0.05). Meanwhile, after intervention the systolic blood pressure of study group [(131.46 ± 12.89) mmHg] was lower than that of control group [(133.27 ± 12.7) mmHg] and the intervention was significant (t=2.1, P<0.05). Conclusion Long-term use of wearable devices help family physicians improve the effect of health education, consequently the community hospitals are responsible to enhance the input and management of the informatization of family physicians' service, and assist them to broaden the form and content of health education.

Objective To analyze the clinical significance and features of patient s' with inconsistent antineutrophil cytoplasmic antibodies (ANCA) test results between indirect immune fluorescence (IIF) and enzyme-linked immunosorbent assay (ELISA).Methods ANCA were detected with IIF and ELISA method jointly among 12 386 in-patients and a retrospective analysis on the proportion of clinical features and significance was made in 589 cases with inconsistent results using Microsoft Excel 2007 statistical software.Results Among the 589 patients,68 (11.5％) were diagnosed as vasculitis,in which 51 cases as ANCA-associated vasculitis,and 521(88.4％) were diagnosed as non-vasculitis including 181 connective disease and 340 non-connective diseases in which hypertension and cardiopathy were common.The common inconsistent results of ANCA were p-ANCA/ELISA (-),IIF (-)/anti-MPO (+),IIF (-)/anti-PR3 (+),IIF (-)/anti-PR3 (+) anti-MPO (+) accounted for 24.4％ (144/589),29.5％ (174/589),15.9％ (94/589),18.5％ (108/589) respectively,these accounted for 88.3％(520/589) of total inconsistency.Conclusion The spectrum of diseases and clinical characteristics varies widely and often presents with multiorgan involvement in patients with inconsistent ANCA results.These reasons make it easy to be misdiagnosed.Attention should be paid to identify and different these inconsistency.

BACKGROUND: Studies have shown that adipose-derived stem cells have pluripotent differentiation potential, but only 30%-40% of cells can differentiate into mature adipocytes with low adipogenic differentiation potential. Therefore, how to improve the adipogenic differentiation ability of adipose-derived stem cells is a key problem to be solved in the process of soft tissue regeneration. OBJECTIVE: To observe the relationship between the surface marker CD54 of rabbit adipose-derived stem cells and their adipogenic capacity, and to explore the adipogenic differentiation of CD54+/CD54- adipose-derived stem cells underthe same induction. METHODS: We successfully isolated and cultured the adipose-derived stem cells from inguinal subcutaneous fat pads (3 ml) of New Zealand white rabbits, aged 8-12 weeks, which were induced into multi-differentiation and used to detectsurface markers. We sorted the passage 3 adipose-derived stem cells by immunomagnetic beads and divided into two categories including CD54+ and CD54- adipose-derived stem cells. After 14 days of adipogenic induction, the cells in the two groups were subjected to oil red O staining and were compared by detecting the density of mature adipocytes and lipid droplet contenT.RESULTS AND CONCLUSION: The cultured adipose-derived stem cells possessed the characteristics of mesenchymal stem cells that could differentiate into mature adipocytes, osteoblasts and chondrocytes, with CD29, CD44, CD49d, CD54, CD73, CD90 and CD105 positive expression while CD31, CD34 and CD45 negative expression. Fourteen days after adipogenic induction, the density of mature adipocytes and the intracellular lipid droplet content in the CD54+ group were significantly higher than those in the CD54- group (P < 0.05). We also found that the mRNA expressions of PPARγ,ADD1, C/EBPα related to adipogenic differentiation in the CD54+ group were significantly higher than those in the CD54- group (P < 0.05). Taken together, CD54+ adipose-derived stem cells have excellent adipogenic differentiation capacity.

Objective To discuss the variance of working expectation values and actual satisfaction degree in nursing college graduates from various aspects and evaluate their actual career mentality.Methods We investigated the working expectation values and actual satisfaction degree in nursing college graduates who graduated from Chengde medical college in 2004 and 2005.Results The working expectation values and actual satisfaction degree had statistical difference in 5 aspects(P<0.05).We subsequently compared the expectation values between each two groups and drew out the conclusion that these nursing graduates had great requirement of being understood and respected.Conclusion The nursing graduates had higher working expectation values and lower satisfaction degree.They paid more attention to be understood and respected and were in great need of emotional support and mental satisfaction.Hospital administrators should help them accurately orient theirselves,establish career development plan,attach importanee to their mental requirement and adopt effective measures to improve their satisfaction degree and promote the healthy growth of nursing college graduates.

Objective To evaluate the myocardial protective effects of Histidine-TryptophanKetoglutarate (HTK) solution on infants with tetralogy of fallot in cardiac operation through comparison with St.Thomas Ⅱ cardioplegia(STH) and HTK cardioplegia in the operation of tetralogy of fallot.Methods Forty infants with tetralogy of fallot(TOF) were enrolled in this study.Their age ranged from 7 to 35 months,and body mass from 5.3 to 9.5 kg.The infants were randomly divided into HTK (n =20) group and STH (n =20) group who received HTK or STH solution respectively.Then 3 ml blood sample were got at 1,2,4,8,24 and 48 h after the opening of ascending aorta.The serum levels of cardiac troponin Ⅰ(cTnI),creatine kinase(CK) and creatine kinase MB(CK-MB) were measured.Results There was significant difference between two groups in terms of the level of cTnI at different time (F(inner group)=49.94,P＜0.001;F(between group) =10.23,P＜0.001;F (across group) =28.49,P＜0.001),and the level of cTnI in HTK group was lower than that of STH group at 1,2,4,8,24 and 48 h after the opening of ascending aorta (P＜0.05).There was significant difference between two groups in terms of the level of CK at different time (F(inner group) =58.85,P＜0.001;F(between group) =16.43,P＜0.001;F(across group)=18.32,P＜0.001),and the level of CK in HTK group was lower than that of STH group at 1,2,4,8,24 and 48 h after the opening of ascending aorta (P＜0.05).There was significant difference between two groups in terms of the level of CK-MB at different time (F(inner group)=34.51,P ＜0.001;F(between group)=11.03,P＜0.001;F(across group)=10.28,P＜0.001),and the level of CK-MB in HTK group was lower than that of STH group at 1,2,4,8,24 and 48 h after the opening of ascending aorta (P ＜0.05).Conclusion HTK is more valid than STK for improving the ability of anti ischemia of myocardium and cardiac function,reducing arrhythmia and ischemia reperfusion injury on infants with TOF in cardiopulmonary bypass.