The baculovirus expression system has been used extensively for the expression of recombinant proteins in insect cells.Recently,recombinant baculoviruses containing mammalian cell-active promoter element have been used to transduce a broad spectrum of primary and established mammalian cells,including non-hepatic cells.Recombinant baculoviruses have been used successfully for transient or stable gene delivery in mammalian cells in vitro,while the efficiency of delivering gene in vivo is inhibited obviously by complements,but efforts have been made to overcome the problems,for instance,VSV-G-pseudotyped baculoviruses display complement resistance.The mechanism of the transduction is still not clearly understood,though many researches have been done.The baculovirus is able to replicate only in insect cells,but it is incapable of initiating replication cycle in mammalian cells,which guarantees high biosafety of this gene delivery system.In addition,this system is easily manipulated and able to carry large inserts.These attributes will undoubtedly lead to the increased application and continued development of this system for highly effective gene delivery into mammalian cells.

OBJECTIVE: To observe the effects of ginsenoside (Gs) and berberine (Ber), two kinds of active components of traditional Chinese herbal medicine, on transforming growth factor-beta1 (TGF-beta1) and prostaglandin E2 (PGE2) in PG cells. METHODS: Co-culture system of human lung carcinoma cell line PG and human T lymphocyte cell line Jurkat was established. PG cells were treated with Gs (100 microg/ml) and Ber (10 mug/ml) for twenty-four hours, and then cocultured with Jurkat cells. After 24-hour coculture, the state of Jurkat cells was observed with inverted microscope. The viable count of Jurkat cells was detected by trypan blue staining after 6- and 24-hour coculture, and the apoptosis of Jurkat cells was evaluated by flow cytometry. PG cells were treated with 100, 50, 25 microg/ml Gs and 10, 5, 2.5 microg/ml Ber respectively, and the content of TGF-beta1 and PGE(2) in PG cells was detected by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) method. RESULTS: After coculture with PG cells treated with Gs and Ber, the number of Jurkat cells was less than blank control group, and the apoptosis rates of Jurkat cells in Gs- and Ber-treated groups were higher than blank control group. Gs and Ber could promote the secretion of TGF-beta1 in PG cells, but could not change the level of PGE(2). CONCLUSION: Gs and Ber can promote the growth inhibition and apoptosis of Jurkat cells induced by PG cells, which may be related to the up-regulation of Gs and Ber on TGF-beta1 secretion in PG cells.

Purpose To observe therapeutic effect of electroacupuncture Neixiyan (Ex-LE 4 ) and Dubi (ST 35) in treating osteoarthritis of knee joint. Method All the 120 cases were randomly divided into electroacupuncture and control groups, 60 cases in each group,and they were given electroacupuncture and Ritalin slowreleased tablet respectively, and pain, mobility and swelling degree of knee joint were observed before and after treatments. Results In treatment group, the average score increased by 18, while in control group, it increased by 12.33 ( P < 0. 05 ) after treatment. Conclusion Therapeutic effect of electroacupuncture Neixiyan (Ex-LE 4) and Dubi (ST 35) in treating osteoarthritis of knee joint was better than that of administration of Ritalin slow-released tablet.

Objective To analyze and summarize the skills and complications of extracorporeal membrane oxygenation( ECMO) cannu-lation and vein intubation. Methods The clinical data of 21 patients of V-A or V-V ECMO in our hospital from January 2009 to July 2014 were analyzed retrospectively. And the techniques of different catheter sites were summarized. Results Three cases were successfully insert-ed catheter by jugular vein puncture with one time. Four patients with ascending aorta intubation died from uncontrolled severe hemorrhage. Eight peripheral catheter site had a small amount of bleeding,with no more bleeding after pressurized bandage. There were no complications like bleeding, hematoma, hemothorax and pneumothorax in the period of ECMO. Conclusion In the process of the ECMO catheter, the standardized operation could reduce the incidence of serious complicaions including bleeding.

Objective To deepen the understanding of chronic eosinophilic leukemia (CEL).Methods The course of diagnosis and treatment in a case of FIP1L1/PDGFRα fusion gene negative CEL was reported. Flow cytometry was used to analyze the immunophenotype of the cells in peripheral blood and pleural fluid. Karyotype was analyzed with G-banding. The expression of FIP1L1/PDGFRα fusion gene was detected by RT-PCR technique. Routine pathological examination of the tissues from bone marrow, lung and spleen were performed. Result A sixteen-year-old girl had severe anemia, fever, splenomegaly,thrombocytopenia and dominant hypereosinophilia lasting for 22 months. Trephine biopsy showed a hypercellular marrow with eosinophilic proliferation and moderate reticular fibrosis. Eosinophilic infiltration was found in lung and spleen and embolism was also found in spleen. She had a clonal chromosomal abnormality t(5;12)(q31;p13). The expression of FIP1L1/PDGFRα was negative. An abnormal clone of T cells expressing CD3-,CD4-,CD8- was found in peripheral blood and pleural fluid, in which the cional T cell accounted for 5.43% and 1.66% of the total lymphocytes respectively. The patient was refractory to treatment with hydroxyurea, prednisone and interferon alpha. She had poor response to a combination of therapy with low dose cytosine arabinoside, mitoxantrone, vincristine, cyclophosphamide, methotrexate and prednisone. She did not respond to imatinib and died of multiple organ failure. Conclusion The present case fulfilled the WHO diagnostic criteria of FIP1L1/PDGFRα(-) CEL which did not respond to routine treatment and imatinib. Allogenic stem cell transplantation should be considered as early as possible in this case. It is noteworthy that clonal CD3-,CD4-,CD8- T-cell abnormality is related to the pathogenesis of CEL.

Abstract: Objective　To investigate the clinical characteristics and diagnosis key points of brain abscess caused by Nocardia asiatica, and provide a clinical basis for diagnosing and treating intracranial infection caused by Nocardia. Methods　A case of pulmonary Nocardia asiatica complicated with brain abscess diagnosed at the Second Affiliated Hospital of Hainan Medical University was selected to analyze the clinical manifestations, cerebrospinal fluid characteristics, pulmonary and cranial imaging features, and treatment plan, and to summarize the diagnosis and treatment experience. Results　The patient was an elderly woman with a history of diabetes, dry cough was the first symptom without fever or headache. At the beginning of the course, it was diagnosed as pulmonary infection and tuberculosis in the local hospital, and received conventional antimicrobial and anti-tuberculosis therapies, but showed no improvement. The patient developed progressive limb weakness, followed by consciousness disorders, and coma. Cerebrospinal fluid (CSF) adenosine deaminase and lactate dehydrogenase were not abnormal, CSF pressure, protein and white blood cells were high, mainly with multiple nuclear cells. CSF glucose and chloride were normal in the early stage of the disease, but decreased significantly in the later stage. Metagenomic analysis of cerebrospinal fluid indicated Nocardia asiatica with a specific sequence number of 537. Lung CT showed exudation, abscess, and cavity in the right lung. Skull MRI scan + enhancement suggested multiple scattered abscesses in both cerebral hemispheres. The abscesses were of different sizes and showed ring enhancement, with extensive surrounding edema, and ventricular compression. After treatment with meropenem, linezolid, and compound sulfamethoxazole tablets, the cerebrospinal fluid recovered, and the lesions in the lungs and intracranial structures improved. Conclusions　Brain abscess caused by Nocardia asiatica is similar to the tuberculous brain in clinical symptoms, cerebrospinal fluid examination, craniocerebral imaging, so we should be alert to the possibility of Nocardia infection in patients with diabetes. At the same time, metagenomic testing of the cerebrospinal fluid can help confirm the diagnosis. The mortality and disability rates of brain abscess caused by Nocardia are high. Early diagnosis and treatment can improve the prognosis.

Objective To compare the differences and consistency of three criteria of metabolic syndrome(MS)including the third Report of the Adult Treatment Panel Ⅲ of National Cholesterol Education Program(NCEP-ATPⅢ),Chinese Diabetes Society(CDS)and International Diabetes Federation(IDF)used for evaluating in children and adolescents.Methods Four hundred and thirty-two overweight or obesity children and adolescents from Chuansha district of Pudong New Area of Shanghai were selected.Data of investigation of the risk factors of cardiovascular disease were reviewed.Anthropometric measurements and fast plasma glucose(FPG),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C)and dynamic blood pressure monitoring were performed.MS were identified according to NCEP-ATPⅢ criteria modified by Cook,CDS criteria modified by Liang,and IDF criteria,respectively.The consistency of the 3 criteria were studied.Results The number of subjects who met the criteria of MS were 31.0%,9.5% and 25.7% using Cook,Liang and IDF criteria,respectively.The conformity rate between Cook and Liang,Cook and IDF,Liang and IDF were 78.5%,89.1% and 83.8%,and the Kappa value were 0.378 20,0.719 18 and 0.465 37,respectively.Conclusions There are differences in consistency among the 3 criteria of MS.The agreement in the diagnosis of MS using Cook and IDF definition is higher than using Liang and other 2 definitions.The number of subjects who met the Cook criteria of MS is the most.The Cook definition may be more suitable for MS diagnosis in children and adolescents.A high conformity rate between Cook and and IDF criterion is proved.It is necessary to work out a universally accepted criterion adapted to children and adolescents.

Objective To observe the effect of sinomenine (SIN) on the expression of cyclooxygenase (COX2),alpha-7 nicotinic acetylcholine receptor(α7nAChR) and adenosine receptor(A2A) in A549 cells,and to explore the relative mechanism for cell proliferation.Methods The effect of SIN and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) on the proliferation of A549 cells was determined by methyl thiazolyl tetrazolium (MTT) assay.The effect of SIN and NNK on the migration of A549 cells was detected by cell wound scratch assay.The effect of SIN and NNK on COX2 expression in A549 cells was determined by Western blotting method.The effect of SIN and NNK on the expression of α7nAChR and A2A mRNA and protein was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting method.Results NNK increased the proliferation and migration of A549 cells,while SIN inhibited the proliferation and migration of A549 cells.COX2 expression level was increased in NNK group but was decreased in SIN group.The expression levels of α7nAChR and A2A were up-regulated in NNK group but were down-regulated in SIN group.Conclusion SIN plays a role in inhibiting the proliferation and migration of A549 cells by suppressing COX2 expression.SIN has an inhibitory effect on the expression of α7nAChR and A2A.

Objective To find the optimal scanning time window and then set up the normal binding potentials (BPs) of 2β-carbomethoxy-3β-(4-fluorophenyl)-(N-11C-methyl) tropane (11C-CFT) DAT PET/CT imaging.Methods Thirty-one healthy volunteers (20 males,11 females,average age:(55.7±2.3) years),who all gave written informed consent,were divided into three age and gender-matched groups according to block randomization.Each group underwent static PET/CT scan in different time windows from 40-60 min,60-80 min to 80-100 min after 11C-CFT injection.To determine the best scanning time window,the ratios of caudate and putamen of all volunteers were analyzed using automatic ROI method (caudate (putamen)/parieto-occipital cotex-1) and compared by one-way analysis of variance and the least significant difference (LSD) t test.The ratio of the same area between different age-groups and gender-groups was compared with independent two-sample t test.Results Ratios of left caudate (2.08±0.06,1.75±0.07and 1.77±0.12 respectively),right anterior putamen (2.33±0.06,1.95±0.09 and 2.08±0.12 respectively)and bilateral posterior putamen (left:1.88±0.66,1.55±0.88 and 1.72±0.09; right:1.98±0.07,1.61±0.09and 1.69±0.12) were all different in three time windows (F=3.588,3.345,4.479,3.557,all P＜0.05).There were significant differences in ratios of left caudate,right anterior and bilateral posterior putamen between 40-60 min and the 60-80 min (all P＜0.05),as well as the ratios of left caudate between 40-60 min and the 80-100 min group (P＜0.05).While no valid differences in ratios of those areas were shown between the groups of 60-80 min and 80-100 min scanning time window (all P＞0.05).DAT densities in right and left side of caudate,anterior and posterior putamen were significantly lower in the group over 60 years of age than those under 60 years (t=-3.260,-3.090,-3.270,-3.190,-2.270,-3.110,all P＜0.05),but were not different between gender-groups (t =0.367,0.522,0.144,0.524,0.166,0.004,all P＞0.05).Conclusions Sixty min after injection of 11C-CFT,the BPs achieve stable stage,so the best scanning time window of 11C-CFT DAT PET imaging was 60-80 min after injection.Identification of normal DAT distribution and understanding of normal BPs of 11C-CFF are necessary before its clinical application.

Adult ; Aged ; Antigens, CD34 ; metabolism ; Chondrosarcoma, Mesenchymal ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Hemangiopericytoma ; metabolism ; pathology ; radiotherapy ; surgery ; Humans ; Male ; Meningeal Neoplasms ; metabolism ; pathology ; radiotherapy ; surgery ; Meningioma ; metabolism ; pathology ; Middle Aged ; Neoplasm Recurrence, Local ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Solitary Fibrous Tumors ; metabolism ; pathology ; Vimentin ; metabolism ; Young Adult