Objective To analyse the cause for delaying diagnosis of WG and improve the diagnosis level.Methods A retrospective study was done on patients who were admitted to the People's Hospital of Peking University and were diagnosed with WG from 1997 to 2007,by searching the computer system of the hospital for patients with WG,who were misdiagnosed and reported in Chinese Journal of Rheumatology and Clinical Misdiagnosis ﹠ Mistherapy from 2003 to 2007.Results WG included multiple systems and organs ivolvement.The lung,kidney and upper airway were the most commonly involved organs,and ear,eye,external salivary gland and oral cavity were also involved.Most initial symptom and clinical manifestation were not typical manifestation of WG trilogy,which led to misdiagnosis and mistreatment.Anti-neurophil cytoplasmic antibody(ANCN)positive rate was high.The most common pathologic features were necrotizing granulomatous inflammation and vasculitis.Conclusion Clinical manifestation of WG is complicated and it injures multiple system or organs.When the symptoms are atypical,it is easy to lead to misdiagnosis,especially in early stage.We suggest running requisite revision for diagnostic criteria of WG,in order to make early diagnosis and treatment of WG.

Objective To analyze quantitatively the safety and efficacy of statin therapy in acute phrase for acute ischemic stroke with the method of meta-analysis.Methods We performed a systematic literature search including the Cochrane Library,MEDLINE and EMBASE for published trials about statin therapy and the outcomes of acute ischemic stroke.Then we performed a meta-analysis with included studies to investigate the association between statin therapy and clinical outcome and mortality.All of the data were pooled and meta-analyzed by Cochrane Collaboration RevMan 5.3 meta-analysis software.Statistical heterogeneity between studies was evaluated by the chi-square and I-square tests.Forest plots were used to summarize study data and Egger tests were used to assess publication bias.Results A total of 27 studies including 52 034 patients,comprising 19 212 statin users and 32 822 non-statin users met the inclusion criteria,4 studies were randomized controlled trials (RCTs),and 23 were observational trials (OTs).Both pre-or post-stroke statin use was associated with reduced mortality.Statin use is associated with favorable functional outcome at hospital discharge and on the ninetieth day regardless of initiation time for pre-stroke group and post-stroke group.The results from observational trials were consistent with randomized controlled trials.There was no evidence of publication bias for all comparisons by Egger tests.Conclusions Statin therapy before or after AIS is safe and effective.

ObjectiveTo construct human embryonic kidney cells (HEK293) modified with human preproenkephalin (hPPE) gene.MethodshPPE gene fragments were obtained from recombinant plasmid pcDNA3.1( + )/hPPE by using restriction endonuelease Hind Ⅲ and Not Ⅰ.Homologous recombination of lentivirus and hPPE gene was produced by using recombinant DNA technology.HEK293 cells were then transfected with the recombinant lentivirus vectors.The expression of hPPE gene in HEK293 cells was detected by Western blot.ResultsThe results of DNA sequencing indicated that the positive clone of recombinant lentivirus was completely consistent with sequencing of hPPE in Genebank.The titer of the concentrated virus was 2.07 × 108 TU/ml.GFP fluorescence was not seen in HEK293 cells transfected with the lentiviral vector under fluorescence microscope.A strong fluorescence was seen in HEK293 cells transfected with Ubc-GFP-L.V.empty viral vector.Positive expression of hPPE was demonstrated in HEK293 cells transfected with lentiviral vector by Western blot.Conclusion HEK293 cells modified with hPPE gene were successfully constructed and the target gene hPPE was stably expressed in HEK293 cells.

Objective Self-made docetaxel-carrying microbubbles were developed and evaluated as a new ultrasound contrast agent for diagnosis and targeted-chemotherapeutic drug delivery.Methods Docetaxel was mixed with an aqueous suspension of phospholipids in vials,which were put into 40℃ water for 30 minutes,after cooling,gas in vials was replaced with perfluoropropane gas,then vials were agitated for 45 s on a shaking device.Properties were studied contained concentration,size,zeta potential,drug entrapment efficiency and drug-loading amount.Drug released with ultrasound and imaging for VX2 carcinoma in rabbits were observed.Results Docetaxel-carrying microbubbles had a concentration of approximately 2.2×109～3.2×109/ml,a mean size of 623.1 am and a zeta potential of -(3.1±0.9)mV.Size distribution was 473.4～706.6 nm.Drug entrapment efficiency was more than 70% and drug-loading amount was(17.5±0.8)%.Fixed amount of ultrasound energy ruptured microbubbles and released docetaxel.Liver imaging of rabbits could be enhanced obviously and persistently,"fast in and out"phenomena was typical of VX2 carcinoma.Conclusions Liposome microbubbles represent a new class of acoustically active drug delivery vehicles.Docetaxel-carrying microbubbles can promote the value of ultrasound in tumor diagnosis,and docetaxel-carrying microbubbles combined with ultrasound have a hope to establish a kind of real-time monitoring,targeted system for tumor therapy.

Objective To investigate the targeted capacity of ultrasound contrast agent with a galactosepoly-L-lysine ligand,and lay the foundation for targeted imaging in vivo and the treatment of liver cancer.Methods The small molecular targeted ligand was synthetized by a method of reductive amination,the targeted lipid microbubbles were performed by combining lipid microbubbles with the targeted ligand,the targeted effectiveness was observed by fluorescence microscopy.Results Galactose and poly-L-lysine were combined effectively after 24 h with the molar ratio 1:1 00,the average particle size of the targeted lipid microbubbles was 2 micron,the targeted combine of the HepG2 liver cancer cells with the targeted ultrasound contrast agent was observed by laser scanning confoeal microscope.Conclusions The targeted lipid miembubbles can target effectively to the HepG2 cells.

OBJECTIVETo summarize the clinical characteristics of an infant with chronic myelogenous leukemia (CML) and the effects of imatinib on the case.

METHODThe clinical features of an infant with CML, who was treated with imatinib in the Norman Bethune International Peace Hospital at June 2009, were retrospectively analyzed and the reports in literature were reviewed. The 1-year-old boy suffered from recurrent low-degree fever and pallor. He had a moderate anemia, distended abdomen and marked splenomegaly. Bone marrow aspiration revealed CML in chronic phase)CP). The t (9; 22))q34; q11) could be detected and BCR-ABL (p210) was positive. The boy was diagnosed as CML-CP and treated with imatinib 100 mg per day. There were 10 related papers and more than 100 child CML patients were reported as retrieved from CNKI)from its establishment to August 2014) and Wanfang Database)from its establishment to August 2014) when "Child", " Chronic" and "Leukemia" were used as keywords. And there were 30 related papers including 400 cases from PubMed Database (from its establishment to August 2014) and one detailed report of an infant with CML was retrieved when "childhood" and "chronic myeloid leukemia" "imatinib" were used as keywords. The clinical effects of imatinib in infant CML cases were analyzed and summarized based on the literature.

RESULTThe boy obtained a complete hematologic response (CHR) at the 6th week of diagnosis, a complete cytogenetic response (CCyR) at the 3rd month and a complete molecular response)CMR) at the 12th month without side effect. This boy grows very well and after a 62-month follow-up, his disease was stable. According to the domestic literature, 5 children CML cases aged 6 -12 years were treated with imatinib without side effects and got complete hematologic response (CHR) after 2-month-therapy. The dose, metabolic characteristics and clinical observation of imatinib can be found in foreign literature and imatinib showed good response with good tolerance in children with CML. Imatinib is regarded as the first line drug for children CML. But it may affect the development of the children.

CONCLUSIONThe children with CML-CP had a good response to imatinib, but more experience in the treatment of children with CML with iniatinib is needed.

Anemia ; Antineoplastic Agents ; therapeutic use ; Fusion Proteins, bcr-abl ; Humans ; Imatinib Mesylate ; therapeutic use ; Infant ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; Male ; Remission Induction ; Retrospective Studies

Objective To evaluate the release of leucine-enkephalin (L-EK) in the cerebrospinal fluid induced by intrathecal human embryonic kidney (HEK293) cells modified with human preproenkephalin (hPPE)gene and the analgesic efficacy of L-EK for bone cancer pain.Methods Forty CIBP female Sprague-Dawley rats were randomized into transplantation (CIBP+ hPPE/HEK293,n =20) and control (CIBP + HEK293,n =20)groups using a random number table.At 1 day before inoculation of cancer cells (T1,baseline) and 8,15,21,25,32 and 35 days after inoculation (T2-7),thermal paw withdrawal latency (TWL) was measured,and the number of licking/biting the claw on the transplantated side and degree of hindlimb limping during free activities were recorded.After observation at T4,10 rats were chosen from each group and sacrificed and the cerebrospinal fluid of rats was collected in an ice bath for detection of hPPE expression using radioimmunoassay.Results Compared with control group,TWL was significantly prolonged,the concentration of L-EK in the cerebrospinal fluid was increased,and the number of licking/biting the claw on the transplantated side and degree of hindlimb limping during free activities were decreased at T4-7 in transplantation group.Conclusion Intrathecal HEK293 cells modified with hPPE gene can continuously secrete L-EK and mitigate bone cancer pain.

Objective To study the effect of mitogen-activated protein kinas/extracellular signalregulated kinase (MAPK/ERK) signaling pathway on cell proliferation modulated by Sonic Hedgehog (Shh) signaling in fibroblast-like synoviocytes (FLS) isolated from patients with active rheumatoid arthritis (RA).Methods The synovial tissue were collected by the synovial arthroscopic debridement or arthroscopic synovectomy of RA patients with active disease activity [disease activity score(DAS)28 ≥3.2].The RA-FLS were primarily cultured by the explanted culture,and then were treated with Shh agonist purmorphamine,inhibitor cyclopamine or MAPK/ERK signaling pathway inhibitor U0126,respectively.Western blotting was used to examine the phosphorylation level of ERK 1/2 (p-ERK1/2),which was the critical protein of MAPK/ERK signaling.The cell proliferation activity was detected using cell proliferation and cytotoxicity kit-8 (CCK8),and the cell proliferation rate was detected using a flow cytometry.Analysis of variance and Kruskal-Wallis H(K) test were used for statistical analysis.Results Compared with the control group,purmorphamine transiently increased p-ERK1/2 protein at the concentration of 1 μmol/L,and the peak activations of p-ERK1/2 took place at 15 min (P＜0.01).Cyclopamine and U0126 decreased the expression ofp-ERK1/2 protein (P＜0.01).After the RA-FLS treated with purmorphmine(1 μmol/L)for 48 hours,the cell proliferation activity was (114±4)％ and the percentage of S phase cells was (8.39±0.60)％,which was significantly higher than those of the control group (100±0)％ (P＜0.01) and (3.29±0.69)％ (P＜0.01).After treated with cyclopamine (10 μmol/L) for 48 hours,the cell proliferation activity of RA-FLS was (89±1)％ (P＜0.05) and the percentage of S phase cells was (1.53±0.22)％ (P＜0.05).When co-treated with purmorphamine (1 μmol/L) and U0126 (10 μmol/L),the cell proliferative activity was (89±2)％ (P＜0.05) and the percentage of S phase cells was(1.07±0.25)％(P＜ 0.05).Conclusion Shh may promote proliferation of RA-FLS via modulating MAPK/ERK signaling,which in turn contributes to hyperplasia of synovium and ultimately leading to RA.

OBJECTIVETo explore clinical value of circular DNA in acute myocardial infarction.

METHODSVenous blood (2 ml/head) of 40 healthy control and 40 patients with acute myocardial infarction within 48h of onset of illness and convalescent period was collected. The level of plasma circular DNA was detected by duplex real-time polymerase chain reaction assay. The levels of myocardial enzyme spectrum and cardiac troponin T (cTnT) were detected by biochemistry method.

RESULTSThe level of circular DNA in control group and group of acute myocardial infarction before treatment was (21.5 +/- 10.7) ng/ml and (253.6 +/- 45.7) ng/ml, respectively (P = 0.000). The levels of serum myocardial enzyme spectrum and cTnT before treatment in patients with acute myocardial infarction were significantly higher than those of control group (P < 0.05). The level of circular DNA after treatment in patients with acute myocardial infarction was significantly decreased compared with that before treatment (P = 0.000), the levels of myocardial enzyme spectrum and cTnT were also significantly reduced (P < 0.05). There was significant correlation between the level of circular DNA and those of CK-MB and cTnT, r = 0.613, 0.654, P = 0.032, 0.021.

CONCLUSIONCircular DNA can be used as a marker of sensitively reflecting myocardial cell injury.

Aged ; Case-Control Studies ; DNA, Circular ; blood ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; blood ; diagnosis

Objective:To explore any effect of electroacupuncture (EA) at the Zusanli point on the scorch death of duodenal cells in rats with functional dyspepsia (FD) and possible mechanisms.Methods:Twenty-four 7-day-old Sprague-Dawley rats were randomly divided into a blank group, a model group and an EA group, each of 8. FD was induced in both the model and EA group rats using iodoacetamide gavage with tail-clip stimulation. After successful modeling the EA group was given acupuncture at the Zusanli point and then connected with a Korean acupuncture point nerve stimulator for 2 weeks. The other 2 groups were not given any intervention. The rats′ body weight was recorded before and after the modeling, as well as 7 and 14 days later. The gastric emptying rate and the small intestine propulsion rate of the three groups were detected right after the EA intervention, and the serum expression levels of interleukin-1β (IL-1β) and interleukin-6 (IL-6) were measured using enzyme-linked immunoassays. Real-time fluorescence quantitative polymerase chain reactions were used to detect the transcription levels of IL-1β and IL-6 in the rats′ duodenums, while western blotting was employed to assess the expression of caspase-1 P20 and dermatin D (GSDMD) in their duodenums.Results:After successful modeling, the average body weight of the rats in the model and EA groups was significantly different from that in blank group, and after 7 and 14 days the average body weight of the former groups was significantly different from that of the blank group, with significant differences between the two groups as well. After the EA intervention significant differences were observed in gastric reside and small intestine propulsion rate between the EA group and the model group, as well as between the model and the blank group. After the intervention, there were significant differences between the blank group and the other two groups in the average expression of IL-1β and IL-6 in serum, IL-1β and IL-6 mRNA in the duodenum, as well as the GSDMD and caspase-1 p20 proteins in the duodenum. There were significant differences between the model and EA groups in all of the above measurements.Conclusions:EA at the Zusanli point can significantly reduce the level of scorch death in the duodenum of FD rats, as well as relieve low-grade duodenal inflammation and the clinical symptoms of FD. Its mechanism may be related to the down-regulation of the expression of caspase-1 P20 and GSDMD-N protein, and of inflammatory factors such as IL-1β and IL-6, relieving low-grade duodenal inflammation.