A large number of documents will be generated in the process of new drug research,including lots of references.These data accumulate into institutional repository.Up to now,the management of document knowledge base in the process of new drug research has not yet been published.The paper uses the reference management software,combined with the document knowledge management system,to analyze and research the management model and method for establishing institutional repository.

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.

Objective To investigate the effect of mitogen-activated protein kinases (MAPKs) activation on the heat stressinduced apoptosis of pulmonary microvascular endothelial cells (PMVECs).Methods A mouse model of severe heat stroke was made and TUNEL and immunohistochemistry were employed to detect lung tissue damage.MACS separation was used for isolation of neonatal PMVECs,and TUNEL was utilized to detect the apoptosis of PMVECs.Western blotting was used for determining the MAPKs activation during heat stress recovery (0,2,6h).The monolayer permeability of endothelial cells was detected in terms of transmembrane resistance (TEER) and horseradish peroxidase (HRP).Cells were pretreated with MAPKs activation inhibitors to examine the effect of heat stress on the monolayer cell permeability and apoptosis.Results In mice with severe heat stroke,extensive apoptosis of PMVECs was found in their pulmonary tissues.TUNEL revealed that the number of apoptotic cells increased over time during heat stress recovery period and heat stress could activate MAPKs in PMVECs.Compared with heat stress group,in the cells pretreated with p38 or ERK activation inhibitor PD98059 and SB203580,the monolayer permeability and apoptosis increased while in cells pretreated withJNK inhibitor SP600125,the cellular permeability and apoptosis decreased.Conclusion In mice with severe heat stoke,PMVECs might experience apoptosis and p38 and ERK could inhibit apoptosis while JNK could promote apoptosis.

The aim of this paper is to compare the cytotoxicity and cellular uptake efficiency of three kinds of poly(b-benzyl-L-amino) block-poly(ethylene glycol) nanoparticles (PXA-PEG-NPs) using Calu-3 cells, and select one as a nasal drug delivery vector for curcumin (Cur). Poly(gamma-benzyl-L-glutamate) block-poly(ethylene glycol) nanoparticles (PBLG-PEG-NPs), poly(gamma-benzyl-L-lysine) block-poly(ethyleneglycol) nanoparticles (PZLL-PEG-NPs) and poly(gamma-benzyl-L-aspartate) block-poly(ethylene glycol) nanoparticles (PBLA-PEG-NPs) were prepared by emulsion-solvent evaporation method. MTT assays were used to evaluate the cytotoxicity of PXA-PEG-NPs against Calu-3 cells. The cellular uptake of nanoparticles was visualized by an inverted fluorescence microscope and quantified by a flow cytometer. The results indicated that even at high concentration of 2 mg x mL(-1) the three nanoparticles had no cytotoxicity on Calu-3 cells. Compared to the curcumin solution, the three curcumin-loaded PXA-PEG-NPs showed significantly higher cellular uptake efficiency on Calu-3 cells (at equal concentration of curcumin with 5 microg x mL(-1) Cur solution), PBLG-PEG-NPs group was the highest. The cellular uptake increased with incubation time, and has positive correlation with nanoparticle concentration. In brief, PXA-PEG-NPs are conducive to delivery Cur into cells, and PBLG-PEG-NPs might be provided as a good nasal drug delivery carrier.
Adenocarcinoma ; metabolism ; pathology ; Administration, Intranasal ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; metabolism ; Aspartic Acid ; chemistry ; toxicity ; Cell Line, Tumor ; Cell Survival ; drug effects ; Curcumin ; administration & dosage ; metabolism ; Drug Carriers ; Ethylene Glycol ; chemistry ; toxicity ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Lysine ; chemistry ; toxicity ; Nanoparticles ; Particle Size ; Polyethylene Glycols ; chemistry ; toxicity ; Polyglutamic Acid ; analogs & derivatives ; chemistry ; toxicity

OBJECTIVETo explore the clinical efficacy of the percutaneous kyphoplasty for the treatment of osteoporotic Kummell's disease.

METHODSFrom May 2010 to February 2012, 8 patients with osteoporotic Kummell's disease were treated with percutaneous balloon kyphoplasty. There were 2 males and 6 females, with a mean age of 73.4 years. All the patients suffered from lower back pain for 4.7 months, which affected seriously the patient's quality of life. The anterior vertebral height and Cobb angel was measured on standing lateral radiograph at pre- and post-operatively (2 days after operation) and during the final follow-up. Visual analog scale (VAS), Japanese Orthopedic Association (JOA) and Oswestry disability index (ODI) were used to evaluate pain and function.

RESULTSIncision of all patients healed normally without infection. The level of back pain decreased remarkably after surgery. The mean time of ambulation was 4.3 days (ranged, 1 to 10 days). Cement leakage occurred in 1 case with no symptom. The anterior vertebral height and Cobb angel of the fractured vertebra recovered respectively from preoperative (30.4 +/- 7.4)% and (31.3 +/- 9.9) degree to (70.3 +/- 3.3)% and (9.1 +/- 3.0) degree at the 2nd day after operation. VAS and JOA scores, ODI improved from preoperative 8.7 +/- 1.2, 12.3 +/- 1.7 and (93.3 +/- 4.6)% to 3.1 +/- 1.1, 24.9 +/- 1.6 and (32.2 +/- 5.4)% respectively at the 2nd day after operation. All the patients were followed up, and the duration ranged from 3 to 24 months, with a mean of 4.7 months. At the latest follow-up, the anterior vertebral height and Cobb angel of the fractured vertebra were (69.9 +/- 3.2)% and (10.9 +/- 2.4) degree; the VAS and JOA scores and ODI were 2.2 +/- 1.0, 26.4 +/- 1.4 and (29.2 +/- 4.5)% respectively, which had no obvious difference compared to those results at the 2nd day after operation.

CONCLUSIONBalloon kyphoplasty is safe and effective treatment for osteoporotic Kummell's disease.

Aged ; Aged, 80 and over ; Female ; Humans ; Kyphoplasty ; methods ; Male ; Osteoporosis ; surgery ; Treatment Outcome

BACKGROUND:Schwann cells are important celllines in the process of repairing peripheral nerve injury, and human amniotic homogenate supernatant is shown to secrete a variety of cytokines, which could promote the proliferation of Schwann cells.
OBJECTIVE:To investigate the effect of different concentrations of human amniotic homogenate supernatant on the proliferation of rat Schwann cell96.
METHODS:Schwann cell96 was cultured with high-glucose DMEM containing 20%fetal bovine serum, and the second generation of Schwann cell96 was applied for experiments. The cultured cells were divided into five groups according to different volume fractions of human amniotic homogenate supernatant (0%, 10%, 15%, 20%, 25%) in the medium.
RESULTS AND CONCLUSION:The total protein concentration of human amniotic homogenate supernatant was 675μg/mL, in which the concentration of epidermal growth factor, basic fibroblast growth factor and vascular endothelial growth factor were respectively (470.625±2.546), (4.121±0.026) and (0.172±0.002) ng/L. At 1-7 days, the cellproliferation rate of the 10%and 15%concentration groups was greater than that in 20%and 25%concentration groups (P<0.05);10%and 15%concentrations promoted cellproliferation, while 20%and 25%concentrations inhibited cellproliferation. There were no significant difference in the viability of Schwann cell96 between the control group and the experimental group (P>0.05). Low concentrations (10%, 15%) of human amniotic homogenate supernatant promote the proliferation of Schwann cell96, while high concentrations (20%, 25%) of human amniotic homogenate supernatant inhibit cellproliferation.

Objective To establish and optimize a sensitive and specific quantitative realtime polymerase chain reaction(PCR)method for detection of hepatitis B virus covalently closed circular DNA(HBV cccDNA)in liver tissue. Methods Specific primers and probes were designed to detect HBV DNA(tDNA)and cccDNA. A series of plasmids(3.44 × 100-3.44 × 109 copies/μl)containing a full double-stranded copies of HBV genome(genotype C)were used to establish the standard curve of real-time PCR. Liver samples of 33 patients with HBV related hepatocellular carcinoma(HCC), 13 Chronic hepatitis B patients(CHB)and 10 non-HBV patients were collected to verify the sensitivity and specificity of the assay. A fraction of extracted DNA was digested with a Plasmid-Safe ATP-dependent Dnase(PSAD)for HBV cccDNA detection and the remaining was used for tDNA and β-globin detection. The amount(copies/cell)of HBV cccDNA and tDNA were measured by a real-time PCR, using β-globin housekeeping gene as a quantitation standard. Results The standard curves of real-time PCR with a linear range of 3.44 × 100 to 3.44 × 109 copies/μl were established for detecting HBV cccDNA and tDNA, and both of the lowest detection limits of HBV cccDNA and tDNA were 3.44 × 100 copies/μl. The lowest quantitation levels of HBV cccDNA in liver tissues tested in 33 HBV related HCC patients and 13 CHB patients were 0.003 copies/cell and 0.031copies/cell, respectively. HBV cccDNA and tDNA in liver tissue of 10 non-HBV patient appeared to be negative. The true positive rate was increasing through the digestion of HBV DNA by PSAD, and the analytic specificity of cccDNA detection improved by 7.24 × 102 times. Liver tissues of 2 patients were retested 5 times in the PCR for detecting cccDNA and the coefficience of variations on cycle threshold (Ct)were between 0.224%-0.609%. Conclusion A highly sensitive and specific quantitative real time PCR method for the detection of HBV cccDNA in liver tissue was established and could be used for clinical and epidemiological studies.

Objective To study the contents of puerarin in decoction with different compatibility ratios of Astragali Radix and Puerariae Lobatae Radix; To verify the rationality of compatibility ratios of Huangqi Gegen Decoction and Yu y e Decoction; To provide references for clinical medication. Methods Different compatibility ratios of Astragali Radix and Puerariae Lobatae Radix were set as 0:1, 1:1, 2:1, 3:1, 4:1, 1:2, 1:3, and 1:4. SinoChrom ODS-BP (4.6 mm × 250 mm, 5 μm) was used as the chromatographic column to detect the contents of puerarin;methanol-0.1% phosphoric acid (35:65) was used as the mobile phase with 0.5 mL/min flow rate; sample volume was 20 μL; the wavelength was 250 nm. Results The regression equation of puerarin was Y=66 449.269 1X-175 665.663 1 (r=0.999 6) in the range of 5 to 300 μg/mL, showing a good linear relationship. The repeatability, stability and recovery rate were fine as well. The contents of puerarin were (2.506 7±0.025 8)%, (2.526 7±0.071 2)%, (2.863 3± 0.086 4)%, (2.956 7±0.119 6)%, (2.835 0±0.078 7)%, (2.480 0±0.072 4)%, (2.530 0±0.064 8)%, and (2.183 3±0.128 9)% in different compatibility ratios of Astragali Radix and Puerariae Lobatae Radix with 0:1, 1:1, 2:1, 3:1, 4:1, 1:2, 1:3, and 1:4, respectively. Conclusion When the compatibility ratios of Astragali Radix and Puerariae Lobatae Radix are 2:1. 3:1, and 4:1, the contents of puerarin in decoction are the highest. This study verifies that the compatibility ratios of Astragali Radix and Puerariae Lobatae Radix in classical prescriptions are rational, which can be the optimal compatibility ratios in clinic.

ONYX-015 and H101 are E1B 55-kDa protein-deficient replicating C group adenoviruses that are currently in clinical trials as antitumor agents. However, their application in cancer gene therapy is limited by the native tropism of C group adenoviruses. This is in part due to low expression of the C group adenovirus receptor (coxsackievirus-adenovirus receptor, CAR) on malignant tumors. An H101-based chimeric virus vector containing sequences encoding the Ad35 fiber domain instead of the Ad5 fiber (H101-F35) was constructed. This modification allowed infection of tumor cells through CD46, a membrane protein over-expressed on tumors. The CAR and CD46 RNA expression was evaluated by RT-PCR method. H101-F35 conferred a stronger cytocidal effect than H101 and ONYX-015 in tumor cell lines that lacked CAR expression (MDA-MB-435 and MCF-7), while the cytocidal effect of H101-35, H101 and ONYX-015 was similar in high-level CAR expressing cancer cell lines (A549, NCI-H446, Hep3B, LNCaP, ZR-75-30 and Bcap-37). In an MDA-MB-435 xenograft mouse tumor model, tumor growth in mice receiving H101-F35 was significantly inhibited compared with mice injected with H101. These results suggest that the chimeric oncolytic adenovirus H101-F35 vector might be a useful candidate for gene therapy of cancer.

OBJECTIVETo compare therapeutic effects of local injection with autoallergic platelet rich plasma (PRP) or autoallergic whole blood (AWB) for the treatment of chronic tennis elbow.

METHODSFrom January 2011 to January 2014, 40 patients with chronic tennis elbow were divided into 2 groups, 20 cases in each group: PRP group and AWB group. There were 20 patients in PRP group treated with local injection of autoallergic platelet rich plasma, including 5 males and 15 females, with an average age of (47.50 ± 9.86) years old; and the average course of disease was (4.67 ± 3.27) months. Among the 20 patients in AWB group treated with local injection of autoallergic whole blood, 3 patients were male and 17 patients were female, with an average age of (46.50 ± 9.96) years old;and the average course of disease was (4.53 ± 2.27) months. The elbow joint was fixed with elastic stockings after injection. All the patients were guided to do strengthening and extension exercises during the follow-up period. Visual analog scale (VAS), Mayo scores for elbow and pressure pain threshold (PPT) were used to evaluate clinical effects after injection immediately and 4,8 weeks after treatment. Results:All the patients were followed up,there were no infections and swelling occurred. The VAS, Mayo and PPT scores of patients in PRP group were improved from pre-therapy 7.22 ± 1.32, 56.71 ± 10.90 and 17.47 ± 4.62 to 2.73 ± 1.00, 91.59 ± 6.95 and 21.35 ± 4.80 respectively 8 weeks after treatment. The VAS, Mayo and PPT scores of patients in AWB group were improved from pre-therapy 7.16 ±1.27, 54.72 ± 8.36 and 17.06 ± 4.83 to 3.81 ± 1.36, 82.06 ± 7.89 and 20.12 ± 4.97 respectively 8 weeks after treatment. All the pain and functional variables including VAS, PPT, and Mayo scores were improved significantly in both groups 4 weeks after injection. On the 4th week after injection, there was no statistically significant difference in PPT between two groups; while the VAS and Mayo score of AWB group were lower than those of PRP group. On the 8th week after injection, the VAS of AWB group was higher than that of PRP group; but the Mayo and PPT scores of AWB group were lower than those of PRP group.

CONCLUSIONPRP and AWB injections are both effective to treat chronic lateral epicondylitis. Compared with AWB injection, PRP injection may be more effective in releasing pain and improving function for a longer time.

Adult ; Blood Transfusion ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Pain Threshold ; Platelet-Rich Plasma ; Tennis Elbow ; therapy ; Visual Analog Scale