OBJECTIVE:To provide reference for rational drug use in pediatric department. METHODS:The utilization of pe-diatric prescription drugs in inpatient department of our hospital during 2008 to 2014 was analyzed in respects of number,consump-tion sum,DDDs,etc. RESULTS:The consumption sum of antimicrobial drugs increased from 657 000 yuan in 2008 to 1 453 000 yuan in 2014. The consumption sum of creatine phosphate sodium increased from 384 000 yuan in 2012 to 889 000 yuan in 2014. The consumption sum of Lysine hydrochloride and sodium chloride injection entered the top 5 in 2014,reaching 205 000 yuan. The consumption sum of essential medicines changed greatly due to varieties. Top 5 antimicrobial drugs in the list of DDDs mainly wereβ-lactam,showing a descreasing trend on the whole. CONCLUSIONS:There are some problems in use of part drugs. It is need to strengthen propaganda and intervention of rational drug use.

Objective:To evaluate the safety of the combination of cefotiam sodium and furosemide and to provide the reference for clinical medication. Methods:Three groups were established, including the blank control group, cefotiam sodium group at the dosage of 500 mg·kg-1 ·d-1 , cefotiam sodium combined with furosemide group at the respective dosage of 500 mg·kg-1 ·d-1 and 15 mg ·kg-1 ·d-1 . After the continuous administration for 12 days, the renal structure, serum uric acid, creatinine and urea nitrogen, u-rine of α1 microglobulin and β2 microglobulin in the rats were detected. Results:Cefotiam sodium at the dosage of 500 mg·kg-1 · d-1 showed no significant effects on the renal structure, serum uric acid, creatinine and urea nitrogen,urineα1 microglobulin andβ2 microglobulin in the rats. The combination group showed significantly increased urine β2 microglobulin (P<0. 05) and significantly decreased serum uric acid (P<0. 05). Conclusion:Short time use of cefotiam sodium exhibits no significant effect on the renal struc-ture and function in rats, while the combination of cefotiam sodium and furosemide has significant effects on urineβ2 microglobulin and serum uric acid in rats.

OBJECTIVETo develop a novel vaccine by immobilizing interleukin-21 (IL-21) on the surface of MB49 cells and evaluate its effect in inducing specific cytotoxic T lymphocytes (CTLs) and antitumor immunity in a mouse model of subcutaneous metastatic bladder cancer.

METHODSSA-IL-21 was immobilized on the surface of 30% ethanol-fixed MB49 cells to prepare the cell vaccine. C57BL/6 mice with subcutaneous implantation of MB49 bladder cancer cells were randomized into 5 groups to receive treatments with IL-21/MB49 vaccine, soluble IL-21, GFP surface-modified MB49 cells, ethanol-fixed MB49 cells, or PBS. The tumor growth and CTL were examined to assess the antitumor efficacy of the vaccine.

RESULTSIL-21 surface-modified MB49 cell vaccine significantly inhibited the tumor growth and generated a long-lasting memory response (P<0.05). At the same effector-target (E:T) ratio, the specific CTLs induced by IL-21/MB49 vaccine showed the most potent cytotoxicity against MB49 cells (P<0.05).

CONCLUSIONWith the protein-anchor technique, IL-21 can be efficiently immobilized on the surface of MB49 cells to prepare IL-21/MB49 cells vaccine. The novel vaccine can maintain its biological activity and significantly enhance the cytotoxicity of CTLs against bladder cancer cells.

Amino Acid Motifs ; Animals ; Cancer Vaccines ; therapeutic use ; Cell Line, Tumor ; Female ; Immunotherapy ; Interleukins ; immunology ; Lymphocyte Activation ; Mice ; Mice, Inbred C57BL ; Neoplasm Metastasis ; Neoplasms, Experimental ; therapy ; T-Lymphocytes, Cytotoxic ; immunology ; Urinary Bladder Neoplasms ; therapy

OBJECTIVETo construct full-length human bladder cancer-specific antibody libraries for efficient display of full-length antibodies on the surface of mammalian cells.

METHODSThe total RNA was isolated from peripheral blood mononuclear cells from patients with bladder cancer. The repertoires of IgG1 heavy chain variable region (VH) and Kappa light chain were amplified by RT-PCR using specific primers. The antibody genes were inserted into the vector pDGB-HC-TM to construct the bladder-cancer-specific antibody libraries of heavy chains and light chains. Ten clones from each library were randomly picked for gene sequencing and transient transfection into FCHO cells to analyze antibody display on mammalian cell surface by flow cytometry after staining with corresponding fluorescent labeled antibodies.

RESULTSThe libraries of bladder-cancer-specific antibody heavy chain (IgG1) and light chain (LCk) were successfully constructed. Seven out of the 10 clones randomly selected from the heavy chain library and 9 out of the 10 clones from the light chain library showed correct open reading frame, coding for 7 unique VH and 9 unique LCk. The combinatory library size reached 3.32×10(11).

CONCLUSIONWe have successfully constructed a full-length human bladder-cancer-specific antibody library with a combinatory diversity of 3.32×10(11) based on mammalian display technology, which can be used for screening monoclonal antibodies against bladder-cancer-associated antigens.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; Cell Surface Display Techniques ; Gene Library ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin kappa-Chains ; genetics ; Peptide Library ; Urinary Bladder Neoplasms ; genetics ; immunology

OBJECTIVETo construct a personalized full-length fully human antibody mammalian display library for children with systemic lupus erythematosus (SLE).

METHODSThe total RNA was isolated from the PBMCs of SLE children. The heavy chain variable region and kappa light chain (VH and LCκ) of the antibody genes were amplified by RT-PCR and inserted into the pDGB-HC-TM vector separately to construct the heavy chain and light chain libraries. The library DNAs were transfected into 293T cells and the expression of full-length fully human antibody on the surface of 293T cells was analyzed by flow cytometry.

RESULTSUsing 0.8 µg total RNA as the template, the VH and LCκ were amplified and the full-length fully human antibody mammalian display library was constructed. The VH and LCκ gene libraries had a size of 9.4×10(4) and 8.4×10(4), respectively. Sequence analysis of 10 clones randomly selected from the VH and LCκ gene libraries each showed that 8 heavy chain clones and 7 light chain clones contained correct open reading frames, and flow cytometry demonstrated that all the 15 clones express full-length antibodies on 293T cell surfaces. 293T cells co-transfected with the VH and LCκ gene libraries expressed the full-length antibodies on the cell surface.

CONCLUSIONThe personalized full-length fully human antibody library for SLE children constructed allows display of the full-length antibodies on mammalian cell surfaces, thus providing a valuable platform for analyzing the autoantibodies, their etiological role, and their clinical implications in SLE.

Amino Acid Sequence ; Child ; Gene Library ; Genetic Vectors ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin kappa-Chains ; genetics ; Lupus Erythematosus, Systemic ; genetics ; immunology ; Membrane Proteins ; genetics

OBJECTIVE：To study the effect s of Bifidobacterium combined with L-carnitine on intestinal flora of dysbacteriosis diarrhea model rats. METHODS ：Totally 30 SD rats were randomly divided into blank control group ，model group ，probiotics group（Bifidobacterium triple viable enteric coated capsules 70 mg/mL），L-carnitine group （L-carnitine injection 50 mg/mL）and L-carnitine+probiotics group （L-carnitine injection 50 mg/mL+Bifidobacterium triple viable enteric coated capsules 70 mg/mL）. Except for blank control group ，the rats in other groups were given 50 mg/mL clindamycin phosphate intragastrically （2 mL/rat， once a day ，for 4 consecutive days ）to establish the model of dysbacteriosis diarrhea. On the 5th day of the experiment ，the rats in administration groups were given corresponding drugs intragastrically ，blank control group and model group were given equal volume of normal saline intragastrically ；with the dosage volume of 1 mL/rat，once a day ，for consecutive 7 days. The general situation of rats in each group was observed during the experiment. The feces of normal control group and model group at the end of the modeling and the feces of the rats in administration group after the last administration were collected for genomic DNA extraction，polymerase chain reaction amplification ，library construction and high-throughput sequencing. After processing ，the effective data were analyzed by operational taxonomic unitsclustering and species annotation ，as well as Alpha and Beta diversity of compared with blank control group ，grade 1 feces and grade 2feces were found in model group. The diversity and richness of intestinal flora ，the ratio of Firmicutes/Bacteroidetes and zhongjuanwang7@163.com the abundance of probiotics such as Lactobacillus， Bifidobacterium and Ackermann were significantly decreased （P＜0.05），while the abundance of pathogenic bacteria such as Enterococcus was significantly increased （P＜0.05）. At the end of the recovery period ，compared with model group ，the activity，fecal morphology and color of rats in probiotics group ，L-carnitine group and L-carnitine+probiotics group returned to normal，and the diversity and richness of intestinal flora had no significant difference （P＞0.05）. However ，the abundance of Lactobacillus in intestinal tract was increased to a certain extent ，and the abundance of Ackermann in intestinal tract of rats in L-carnitine+probiotics group was significantly increased （P＜0.05）. CONCLUSIONS ：Although Bifidobacterium combined with L-carnitine have no significant effect on improving the diversity and richness of intestinal flora in dysbacteriosis diarrhea model rats，it could increase the abundance of probiotics to a certain extent.