This paper points out the importance of intravenous thrombolysis with recombinant tissue plasminogen activator(r-tPA)according to the international guidelines in acute ischemic stroke.Some practical problems,such as the lower rate of thrombolysis for ischemic stroke,the price of r-tPA being too expensive to clinical using,the dosage of r-tPA in Chinese being not clear,the curative effect being not so good,and traditional Chinese medicine (TCM) applied in intravenous thrombolysis,and so on,should be further discussed.

Objective To evaluate in vivo tracking of swine mesenchymal stem cells (MSCs) la-beled with super paramagnetic iron oxide (SPIO) in intraportal transplantation by a clinical 1.5T MR.Methods MSCs were isolated from swine and cultured as well as expanded, which were then incuba-ted with SPIO (Feridex I. V.). Prussian blue staining was performed for showing intracelluar irons.To establish a swine model of acute liver necrosis, 0.5 g/kg of D-galactosamine was administrated to 10 pigs. MSCs(labeled cells in six, unlabeled cells in four)were injected into liver via portal veins. MR imaging was performed with a clinical 1.5T MR immediately before and at 6 h, 3 d, 7 d, 14 d after transplantation, respectively. Results Prussian blue staining of SPIO labeled MSCs could be effec-tively labeled and the labeling efficiency was almost 100%. Signal intensity loss in liver by SPIO labe-ling on FFE sequence persisted until 14 days after transplantation. Histological analysis by Prussian blue staining showed homing of labeled MSCs in liver after 14 days, primarily distributing in hepatic sinusoids and liver parenchyma. Conclusion MSCs can be labeled with SPIO in vitro successfully.MRI can monitor magnetically labeled MSCs transplanted into liver.

ObjectiveTo explore the diagnostic value of plasma (1, 3)-β-D-glucan test (G test) in diagnosis of invasive fungal infections (IFI) and the influence of albumin on G test.Methods A prospective observational study was conducted. 267 patients admitted to medical intensive care unit (MICU) of Dalian Municipal Central Hospital from January 21st, 2012 to October 31st, 2014 were enrolled. According to IFI guideline, the patients were divided into without IFI group (n= 35), possible IFI group (n = 70), hypotheticle IFI group (n = 145) and proven IFI group (n = 17). G test was examined routinely using microbiology kinetic rapid reader MB-80.The different threshold values were calculated on G test. The difference among G tests, fungal culture and clinical diagnosis were compared. The results of G test ahead of and post albumin administration in each group were compared, and the value of G test for diagnosis of IFI during albumin infusion was evaluated.Results When the cut-off value was 20 ng/L for IFI diagnosis, higher sensitivity (79.8%), specificity (87.9%), and Youden index (67.7%) were found. The positive rates of G test, fungal culture and clinical diagnosis of IFI were 57.7% (154/267), 60.7% (162/267) and 54.3%(145/267) respectively, without showing significant differences (allP> 0.05). The result of G test (ng/L) was not obviously changed after albumin administration compared with that before in without IFI, possible IFI, hypotheticle IFI, and proven IFI groups (without IFI group: 11.25±2.33 vs. 10.99±1.07,t= -1.723,P= 0.085; possible IFI group: 53.14±5.53 vs. 49.22±8.11,t= -0.395,P= 0.693; hypotheticle IFI group: 90.30±9.38 vs. 85.41±10.11, t= 710.500,P= 0.860; proven IFI group: 100.98±19.24 vs. 103.21±17.66,t= 653.000,P= 0.449). Prior to the administration of albumin, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and Youden index were 79.8%, 87.9%, 45.6%, 96.7%, 67.7%, respectively. However, after the administration of albumin, they were 81.5%, 85.7%, 44.8%, 96.5%, and 67.2%, respectively, without significant difference.Conclusions G test is method for early diagnosis of IFI. The sensitivity and specificity are higher with 20 ng/L as the critical value. The result of G test is not interfered by albumin administration.

Objective:To determine the content of cyclohexane, ethyl acetate, methanol, methylene chloride and trichloromethane in rupatadine fumarate by headspace gaschromatography. Methods:A DB-WAXETRR capillary column(30 m × 0. 32 mm,0. 25 μm)was used and the carrier gas was nitrogen. The detector was an FID and the inlet temperature was 200℃ . The column temperature program was with the initial temperature of 35℃,maintained 10 min,and then risen to 220℃ with the rate of 20℃·min -1 ,and maintained 5 min. Results:Cyclohexane,ethyl acetate,methanol,methylene chloride and trichloromethane showed a good linear relationship within the range of 77. 590 1- 698. 310 9 μg·ml -1(r = 0. 999 7),102. 166 6- 919. 499 4 μg· ml -1(r = 0. 999 8),62. 744 7- 564. 703 2μg·ml -1(r = 0. 999 9),12. 011 2- 108. 101 1 μg·ml-1(r = 0. 999 6)and 1. 262 8-11. 365 6 μg·ml -1(r = 0. 999 6). The average recovery was 103. 9% ,103. 5% ,104. 9% ,107. 1% and 103. 4% and RSD was 2. 3% ,2. 6% ,3. 1% ,2. 8% and 4. 5%(n = 9),respectively. The five residual solvents were not detected out in rupatadine fumarate. Conclusion:The method is stable,simple,sensitive and accurate,and can be used for the determination of residual solvents in rupatadine fumarate.

Objective To explore the clinic effect of micro course in health education for prevention shoulder-hand syndrome in patients with stoke.Method The 42 stoke patients treated in our sector from October 2013 to September 2014 were observed (observation group) under health education by micro course,taking the 45 stoke patients treated in our sector from October 2012 to September 2013 as control group under traditional health education.Heahh education evaluation,occurrence rate of shoulder-hand syndrome,shoulder joint passive range of motion (ROM) and simple test for evaluating hand function (STEF) of the two groups of patients were compared after one and three months,respectively.Results Compared to the control group,the observation group made higher evaluation of health education method,showed lower occurrence rate of shoulder-hand syndrome,better shoulder joint passive ROM and upper limb movement function.In the time point of 1 and 3 months after the intervention,the incidence rate of shoulder-hand syndrome in the observation group was 2.5％(1/40) and 7.5％(3/40) respectively,which was significant lower than that of in the control group (19.05％,8/42 and 47.63％,20/40 respectively),thex2 value was 4.173 and 16.340 respectively,P＜0.05.In the time point of 1 and 3 months after the intervention,the passive ROM in the observation group was (172.80 ± 15.18)° and (174.76 ± 7.45)°,which was significant higher than that of in the control group,(143.81 ± 22.76)° and (132.12 ± 16.67)° respectively,the t value was 6.816 and 15.074respectively,P＜0.01.Conclusion Micro course used in health education of stroke patients could stimulate their study interests and activity,which played an important role in shoulder-hand syndrome occurrence prevention,shoulder injury reduction and upper limb function restoration acceleration.

BACKGROUND: Classic isolation method of bone marrow mesenchymal stem cells (BMSCs) is Percoll density gradient centrifugation. Blood cell component was removed. However, this method is complicated. Preparation density was needed when isolating dog bone marrow. Moreover, centrifugation was frequent, which had a great damage to cells. OBJECTIVE: To establish methods of the isolation, proliferation, culture and osteoinduction of canine BMSCs, and observe the in vitro proliferation and ability to osteoinduction differentiation. METHODS: 10 mL bone marrow was extracted from dog posterior superior iliac spine, heparin anticoagulation, diluted using Hanks juice, treated with 1.077 g/mL Ficoll solution 3 mL, and centrifuged at 2 000 r/min for 20 minutes. Karyocytes were absorbed to form white cloudlike layering interface, and then centrifuged twice using DMEM supplemented with fetal bovine serum, incubated at 12×10~4/cm~2 at 37 ℃ in a 5% CO_2 incubator. Following subculture, cells were incubated in DMEM containing dexamethasone, β-sodium phosphoglycerol and ascorbic acid 2-phosphate. Immunocytochemical staining and immunofluorescence staining were utilized to detect osteocalcin, osteopontin and type Ⅰ collagen expression in osteoblasts. Alkaline phosphatase staining and alizarin red staining were performed. RESULTS AND CONCLUSION: 1.077 g/mL Ficoll density gradient centrifugation was used to isolate karyocytes that were significant compared with Percoll solution. Obtained BMSCs had high purity, good growth and the mean doubling time was 24 hours. Following in vitro osteogenic incubation of dog BMSCs, osteocalcin, osteopontin and type Ⅰ collagen showed positive expression. Alkaline phosphatase staining demonstrated bluish-green cytoplasm. Alizarin red staining showed red nodes in extracellular matrix, and could differentiate into osteoblasts in vitro.

ObjectiveTo develop an antibody-array system for multiple detection of antibodies against Japanese B encephalitis virus (JEV),Tick-borne encephalitis virus (TBV),Dengue virus ( DENV ),West Nile virus (WNV),Western equine encephalitis virus (WEEV) and East Equine encephalitis virus (EEEV).MethodsRecombined antigens were spotted on array as capture antigens.Specific antibodies were detected by using a sandwich ELISA format.Rabbit antiserum was employed to select and confirm the specificity of antigens and to optimize the conditions of the assay.The detection efficiency of the system was validated by 40 clinical suspected serum samples and compared with the relative ELISA assays.ResultsEleven recombined antigens were selected as diagnostic antigens with high specificity.Better detection could be achieved when scale of antigen concentrations were within 0.125-0.900 mg/ml and the serum dilutions were 1:100-1:1000.When detecting the 26 clinical suspected TBE serum samples,20 were IgG positive (76.9％),and 17 were IgM positive (65.3％) which was 96.1％ and 84.6％ consistent with the relevant ELLSA tests,the 8 clinical suspected JEV serum samples,4 were IgG positive (50.0％),and 5 were IgM positive (62.0％),which was 86.3％ and 90.1％ consistent with the relevant ELLSA tests.As for the 22 DEN serum samples,13 were IgG positive (60％) and 15 were IgM positive (68％) which was 85％ and 93％ consistent with ELISA.The specificity of the assay was 100％ and the sensitivity was higher than the relative ELISAs.ConclusionThe developed antibody-array is highly specific and reliable,which could be used for the detection of antibodies against the 6 arboviruses.

Objective:To screen the peptide binding to human bladder carcinoma cells specifically by using phage display technology in vivo.Methods: Nude mice were inoculated with bladder carcinoma cells BIU87 for establishing tumor-bearing mice model.The Ph.D.-C7CTM Peptide Library was injected intravenously via tail vein.Then we screened Phage containing exogenous peptides binding to bladder transitional carcinoma cells specifically.The phage peptide homed to the tumor tissues was obtained after 3 rounds screening in vivo.The phage clones affinity to BIU87 were identified by immunohistochemistry and ELISA.The positive peptide was synthetized by chemical methods after sequencing the positive monoclonal phage DNA.The tumor cell specificity of target peptide was identified by confocal laser scanning microscope and flow cytometry.Results:After 3 rounds screening in vivo,enrichment rate of phage was 4.334×102 times.Immunohistochemistry results showed that the dyeing of the tumor tissue had a rising trend following each round of phage screening,while liver had a lot of non-specific binding phage because the phages were metabolized through liver and kid-ney.The 30 phage clones were identified by ELISA and 10 clones had a strong affinity on BIU87 among 24 positive clones.Three amino acid sequences of positive phage clones were obtained.The highest rate of repeat sequences CSSPIGRHC(8/10) named NYZL1 and the FITC-C6-NYZL1 peptide was synthesized.Our results showed that it could bind to bladder carcinoma cells BIU87 specifically.Conclusion:We obtained the small molecular peptide NYZL1 binding to human bladder carcinoma specifically by means of phage display in vivo,which provide a theoretical basis for bladder carcinoma early diagnosis and targeted therapy.

ObjectiveTo evaluate the effect of Rhubarb associated preparations (rhubarb or prescriptions of traditional Chinese medicine including rhubarb) on sepsis patients with acute gastrointestinal dysfunction (AGI).Methods The retrieval of databases from libraries including PubMed, Medline, Cochrane Central Register of Controlled Trials, CNKI, CBMdisc, Wan Fang Database, VIP database were searched to identify randomized controlled trials (RCTs) about Rhubarb associated preparations for treatment of sepsis patients with AGI from the foundation of the various databases to March 2016. And in the mean time, the references of the studies accepted were also retrieved. The retrieving and screening of literatures were performed independently by two researchers, the methodological quality and data extraction of the enrolled literatures were assessed by Jadad scale, and Cochrane Collaboration 5.3 software was used to perform Meta analyses to observe the effects of rhubarb associated preparations on gastrointestinal function score, acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ) score and 28-day mortality in sepsis patients with AGI; the bias of published literatures was evaluated by funnel plot.Results ① Finally, 16 studies involving 1 171 patients (610 in rhubarb preparation group and 561 in the control group) were identified and enrolled. 12 studies had a Jadad score ≥ 3 and 4 studies < 3. The random method was used in classification of groups in all the studies in which the intergroup baseline data being comparable was clearly indicated. The blind method was applied in 5 contained RCTs.② The results of Meta-analyses showed that rhubarb associated preparation could improve gastrointestinal function score [mean difference (MD) = -0.52, 95% confidence interval (95%CI) = -0.55 to -0.48, P < 0.000 01], reduce the APACHEⅡ score (MD = -3.66, 95%CI = -5.00 to -2.33,P < 0.000 01) and 28-day mortality [odds ratio (OR) = 0.46, 95%CI = 0.30 to 0.71,P < 0.000 01] compared with those in the control group, the differences being statistically significant. No publication bias was seen in 16 literatures containing RCTs from the funnel plot.Conclusions Compared with the control group, the rhubarb associated preparations combined with conventional theraph can significantly improve the gastrointestinal function score, reduce APACHE Ⅱ score and 28-day mortality of sepsis patients with AGI, which suggests the rhubarb associated preparations have better efficacy. In addition, the result of sensitivity analysis has not substantially changed the results of Meta-analysis.

Objective To analyze the positive rate of specific distribution characteristics in Rh blood group antibody,analyze the clinical significance of Rh blood group antibody and rules.Methods The micro column gel anti globulin technique was used to screen and identify irregular red blood cell antibodies,for patients with Rh blood group antibody,monoclonal anti-D,anti-C,anti-c, anti-E,anti-e were used to identify Rh blood group antigen to confirm the accuracy of detection.The antibody titer,Ig-type and 37℃ reactive were used to determine its clinical significance.Through asking pregnancy history,history of blood transfusion,under-standing whether the same specificity of the antibody in maternal plasma if the patient was newborn,the causes of antibody were an-alyzed.Results In 109 000 patients,Rh blood group antibodies were detected in 96 cases,the positive rate was 0.088%,which has a history of pregnancy in 68 cases,5 cases had history of blood transfusion,both pregnancy history and history of blood transfusion in 6 cases,1 7 cases of neonatal maternally derived antibody.Antibody specificity:65 cases of anti-E(67.710%),12 cases of anti-cE (12.500%),8 cases of anti-D (8.330%),7 cases of anti-c(7.291%),2 cases of anti-C (2.083%),2 cases of anti-e(2.083%).96 cases of Rh blood group antibodies were IgG or IgG+IgM class,37 ℃ reaction could be with the corresponding antigen of red blood cell,antibody titer between 4-2 048.Conclusion Anti-D detection rate shows a trend of gradually decreasing.In Rh blood group antibody detection,anti-E and anti-cE account for an absolute majority.Alloimmune caused by pregnancies and blood transfusion is the main reason of Rh blood group antibody production from Rh blood group antibody.Neonatal maternal passive getisa Rh-HDN is the main pathogenic antibody.