Objective To investigate the efficient methods used for tracing endothelial progenitor cells(EPCs)after transplantation in injured lung tissue.Methods Thirty clean grade healthy syngeneic SD female rats were divided into three experimental groups(n =10):(1)sham group,rats treated with intravenous phosphate-buffered saline(PBS)instead of lipopolysaccharide(LPS)followed by EPCs graft;(2)PBS-treated group,rats treated with intravenous PBS after intravenous injection of LPS to produce acute lung injury;(3)EPC-treated group,rats treated with EPCs after intravenous injection of LPS to produce acute lung injury.The transplanted EPCs were got from the same genetic species of the SD male rats.Rats of each group were sacrificed 7 days after EPCs transplantation.Their whole lung tissues were harvested to detect the expression of Y-chromosome by using hybridization in situ and RT-PCR assay.Statistic package of SPSS16.0 was used for the data analysis and significant differences between means were evaluated by ANOVA analysis.Results Compared with the other two groups,positive signals of sex-determining region y were found in lung endothelium from the EPC-treated group.Conclusions Y chromosme specific probe can be one of efficient methods for tracing stem cells after transplantation.

Objective To investigate the effect of mild hypothermia preconditioning against ischemia/ reperfusion (I/R) injury of cultured primary cortical rats neurons,and to compare the protective effect of mild hypothermia only and with its combination with drugs.Methods Cortical neurons of neonatal Sprague-Dawley (SD) rat within 24 hours after birth were harvested and cultured in vitro.On the 3rd day,the cells were cultured in a medium containing 2.5 mg/L cytosine arabinoside to inhibit the growth of glial cells and fibroblast.Having cultured for 6 days they were randomly divided into blank control group,glutamate damaged group (cultured with 200 μmol/L glutamate for 0.5 hour after washing),mild hypothermia preconditioning group (cultured under 33.5 ℃ for 24 hours before injury induced by glutamate),mild hypothermia combining with edaravone preconditioning group,and the hypothermia combining with propofol preconditioning group (medium containing 100 μmol/L edaravone and 3 mg/L propofol).They were cultured under 33.5 ℃ for 24 hours before injury induced by glutamate.After 24 hours of culturing in various medium,apoptosis ratio was observed by flow cytometry.Cell surviving rate was determined with methylthiazolete trazolium (MTT),c-fos protein expression was assayed,and morphologic change of cells with hematoxylin-eosin (HE) staining under the microscope,and uhrastructure changes were observed after uranyl acetate and lead citrate staining under transmission electron microscope.Results The apoptosis ratio and c-fos protein in glutamate damaged group were significantly higher than those in blank control group [apoptosis ratio:(9.85 ± 0.76)％ vs.(0.94 ± 0.20)％,c-fos (ng/L):6.96 ± 0.75 vs.1.65 ± 0.59,both P＜0.01],the cell surviving rate was significantly lower than that in blank control group [(0.20 ± 0.02)％ vs.(0.97 ± 0.03)％,P＜0.01].Mild hypothermia preconditioning reversed surviving rate,apoptosis ratio and c-fos protein,and the effect of hypothermia combining with propofol preconditioning was obviously better [cell surviving rate:(0.80 ± 0.04)％ vs.(0.20 ± 0.02)％,apoptosis ratio:(2.26 ± 0.54)％ vs.(9.85 ± 0.76)％,c-fos (ng/L):2.98 ± 0.46 vs.6.96 ± 0.75,all P＜0.01].The morphology of cortical neurons in blank control group was normal.Most of the cells in glutamate damaged group showed bluish black cytoplasm with pyknic nuclei,with crimpled axons and of them were fractured,and cell number was obviously decreased.In each pre-conditional groups,decrease in cell number was unconspicuous,and only a few cells showed apoptosis.Under transmission electron microscope,it was found that cell membrane,mitochondria and rough endoplasmic reticulum were intact in blank control group,but with reduction in organelles,severely swollen mitochondria,even with formation of vacuole or pyknosis,serious dilation of rough endoplasmic reticulum,with loss of cristac with loss of vacuoles or pyknosis,and marked dilatation of intemal reticular endoplasm,and loss of cristac with vacuolation and chromatin were observed under electron microscope in glutamate damaged group.Compared with the glutamate damaged group,these pathologic changes were markedly alleviated in protected groups.Conclusions Mild hypothermia preconditioning can inhibit glutamate-induced injury to cortical neurons.The protective effect of mild hypothermia combined with propofol is better.

Objective To optimize prescription for double-layered Erhuang sustained-release suppository. Methods Amounts of PEG400, PEG4000, HPMC were selected as influence factors for L9(34) orthogonal experiment. A comprehensive assessment was conducted by setting the cumulative release degree at three different time points as index, and the inner and outer layers of double-layered Erhuang sustained-release suppository were optimized. Results The best prescription was the inner HPMC∶PEG4000∶PEG400=1.5∶10∶4;outer HPMC∶PEG4000∶PEG400=0.5∶10∶4. Conclusion Prescription for double-layered Erhuang sustained-release suppository has good forming property and a good sustained-release effect according to the optimized prescription, which has certain reference value for researches and development of TCM suppository.

Objectves To investigate the effectiveness and safety of bronchoalveolar lavage for the treatment of neonatal pulmonary atelectasis under ultrasound monitoring.Methods Thirty-two neonates diagnosed with pulmonary atelectasis by lung ultrasound,and hospitalized in the Neonatal Intensive Care Unit of Bayi Children's Hospital,the General Hospital of the Chinese People's Liberation Army between July 2014 and June 2016,were included in this study.Bronchoalveolar lavage was performed in all the patients by injection of lavage fluid (0.9％ NaC1 or 0.9％ NaCl plus ambroxol hydrochloride and/or exogenous pulmonary surfactant) 1.5 to 3.0 ml via tracheal intubation.Lung ultrasonography was performed immediately after each lavage to reveal the status of lung recruitment.Repeated lavage one to three times made up of one course of treatment.The bronchoalveolar lavage could be performed for one to two courses daily according to the status of atelectasis recovery.Medical records were reviewed to analyze descriptively the effectiveness,side effects and complications of the bronchoalveolar lavage.Results Bronchoalveolar lavage was significantly effective in 25 patients (78.1％) with disappearance of pulmonary atelectasis after one course of treatment;effective in five cases (15.6％) with disappearance or reduction of atelectasis after two or three courses;with a total effectiveness rate of 93.8％(30/32).Bronchoalveolar lavage was ineffective in two cases (6.2％) with no remarkable change in atelectasis after three courses of treatment.Vital signs were stable in all the infants during the bronchoalveolar lavage,and no adverse effects and complications occurred.Conclusions Bronchoalveolar lavage is effective for the treatment of neonatal pulmonary atelectasis under ultrasound monitoring,and it is easy to operate and with no adverse effects and complications,and thus worth of clinical application.

A new fluorescent method was developed based on the ulifloxacin-europium (Ⅲ)-sodium dodecylbenzene sulfonate system for the determination of ulifloxacin,the active metabolite of prulifloxacin.Sodium dodecylbenzene sulfonate formed a ternary complex with ulifloxacin-europium (Ⅲ) and significantly enhanced the characteristic fluorescence of europium (Ⅲ).The enhanced fluorescence intensity showed a good linear relationship with the concentration of ulifloxacin in the range of 5.0× 10 8 -2.0× 10-6M with a detection limit of 2.0× 10-10 M (3σ).This method is rapid and sensitive,and has been successfully applied to the determination of ulifloxacin in human urine and serum samples.

A new fluorescent method was developed based on the ulifloxacin-europium（Ⅲ）-sodium dodecylbenzene sulfonate system for the determination of ulifloxacin,the active metabolite of prulifloxacin.Sodium dodecylbenzene sulfonate formed a ternary complex with ulifloxacin-europium（Ⅲ）and significantly enhanced the characteristic fluorescence of europium（Ⅲ）.The enhanced fluorescence intensity showed a good linear relationship with the concentration of ulifloxacin in the range of 5.0×10^-8-2.0×10^-6M with a detection limit of 2.0×10^-10 M（3σ）.This method is rapid and sensitive,and has been successfully applied to the determination of ulifloxacin in human urine and serum samples.

Objective To explore the differences of the joint function and the patient's satisfaction between two different methods to reconstruct limb length for unilateral Crowe Ⅳ developmental dysplasia of the hip (DDH).Methods The clinical controlled study were used.21 cases with unilateral DDH Crowe Ⅳ treated with total hip arthroplasty were divided into 2 groups randomly.The patients in the first group were reconstructed the limb with the compensatory length and those in the second group were did with the real length.The patients were followed up 10 years.Two independent sample t-test was used to compare these two operations.Harris scores and SF-36 scores were used in the test.Survival analysis with Kaplan-Meier method was involved in the survival of the prosthesis and then test with Log-rank test.X-ray films of different period were compared to confirm the prosthetic loosening.Results Seventeen patients were followed up 8-10 years.There were no significant differences both in Harris score and most of the SF-36 scores at the median follow-up of 10 years.The item mental healthy of SF-36 expressed significant difference.Five patients required revision.There were 10 cases suffered with polyethylene wear,6 cases with severe osteoporosis in greater trochanter,and 3 cases with osteolytic reaction.Conclusion There are no significant differences between the two surgical method in the survival rate of prosthesis and the joint function in the median follow up.The satisfaction of the control groups is lower than the trial groups both at the early stage after the operation and the median follow up.The revision rate between the two groups is similar but the reason is different.

A patient with impaired kidney function after kidney transplantation and received treatment at the First Hospital of Jilin University was retrospective analyzed. The patient was male, 45 years old, and was diagnosed hemolytic uremic syndrome by transplanted kidney biopsy. The patient received cyclosporine A (CsA) as maintenance centered immunosuppression therapy postoperatively. He was admitted because of 1 week acratia followed by 1 day increased serum creatinine level at 1.5 years after transplantation. At 1 day after admission, he was received renal needle biopsy, and underwent 2 days Prednlsolone treatment. After hemolytic-uremic syndrome was diagnosed, CsA was transferred to Tacrolimus (Fk506) with dose of 2 mg/d, and Azathioprine was replaced by mycophenolate, Prednisone was taken orally for 20 mg/d. The function of the transplanted kidney and the change of routine blood tests were observed. After 1 week treatment of the changed Immunosuppression therapy, the function of the transplanted kidney was improved obviously, and the hemoglobin and platelets was decreased during the treatment. The results demonstrated that kidney biopsy is a key method to diagnose hemolytic-uremic syndrome, and adjustment of immunosuppressive agents, replacing CsA with FK506 are effective for postoperative hemolytic-uremic syndrome.

AIM:To observed the protective effect of diminazene aceturate ( DIZE) , an angiotensin-converting enzyme 2 (ACE2) activator, on diabetic nephropathy (DN) rats.METHODS:Male Wistar rats (n=30) were randomly divided into normal control (NC) group, DN group and DIZE group (each group consisted of 10 rats).The rats in DN group and DIZE group were induced by intraperitoneal injection of streptozotocin at dose of 65 mg/kg.After 12 weeks, the rats in DIZE group and DN group received subcutaneous injection of DIZE (15 mg· kg -1 · d-1 ) or vehicle for 4 weeks. The samples of blood and urine were collected at week 16, and ratio of kidney weight to body weight (KW/BW), plasma glucose (GLU), 24 h urinary protein (24UP) and serum creatinine (SCr) were measured.The renal pathological changes in each group were observed by periodic acid-Schiff ( PAS) staining and immunohistochemistry .The levels of AngⅡ and Ang-(1-7) in the plasma, and TGF-β1 and VCAM-1 in the renal tissues were measured by ELISA .The mRNA and protein levels of collagen I and FN were determined by quantification real-time PCR and immunohistochemistry .The effects of DIZE on the expression of ACE2 in DN rats were determined by Western blot .RESULTS:DIZE remarkably increased the expression of ACE2 and Ang-(1-7) in DN rats.Compared with NC group , the GLU, KW/BW, 24UP, SCr, and the ex-pression of collagen I , FN, TGF-β1 and VCAM-1 in DN group and DIZE group were increased .However , after treatment of the DN rats with DIZE, these indicators were decreased except the KW/BW.The GLU showed no significant change . CONCLUSION:DIZE raised the activity of ACE2 and increased the expression of Ang-(1-7), thus alleviating fibrosis and inflammation in the kidney and having therapeutic potential for diabetic nephropathy .

Objective An effective reproducible protocol for complete plant regeneration via somatic embryogenesis has beendeveloped for Herpetospermum pedunculosum,an endangered Tibetan medicinal herb.Methods The cotyledonexplants used in this study were excised from seedlings germinated in vitro.Callus was induced from cotyledonexplants on Murashige and Skoog's medium,supplemented with 2,4-dichlorophenoxyacetic acid(2,4-D,0.1-1.0mg/L)alone or in combination with 6-benzylaminopurine(BA,0.5,1.0,and 2.0 mg/L).Results The calli showeddifferentiation of globular embryos after three weeks of incubation on MS medium supplemented with variouscombinations of BA and NAA.Sixty-two percent of the embryogenic calli produced somatic embryos in MS basalmedium supplemented with BA(1.0 mg/L)+NAA(2.0 mg/L).The addition of KN(0.5 mg/L)to MS mediumcontaining both BA and NAA(2.0 mg/L each)significantly increased the frequency of somatic embryogenesis.Themaximum percentage of embryogenic calli formation was 83%,and globular embryos formed and germinatedsuccessfully in this medium.Then,transferring the regenerated plants from this medium to hormone-free MSmedium will further enhanced the development of the plants,and the healthy plantlets are formed successfullywithin four weeks.The plantlets were transferred to soil to acclimatize under greenhouse conditions and 75%survived.Conclusion Somatic embryogenesis protocol as reported here can play a key role in the propagation andconservation of this endangered species.