Objective To explore the role of Adra1a in regulating the LPS-induced inflammation response in primary hepatocytes of lipopolysaccharide-binding protein knockout(Lbp-/-)mice.Methods Primary hepatocytes were extracted from WT and Lbp-/-mice using a two-step perfusion method,and an inflammation model was established using LPS induction.Expression of Adra1a in primary hepatocytes of Lbp-/-mice was suppressed by administering the inhibitor prazosin and transfection with si-Adra1a.The cells were divided into three groups under inhibitor conditions:control group A,LPS group A,and prazosin group.For siRNA transfection,cells were also divided into groups:control group B,LPS group B,si-NC group,and si-Adra1a group.WT primary hepatocytes were divided into two groups:control group(blank)and LPS group(12 h stimulation).Changes in the Adra1a response to LPS stimulation were verified by Western blot.Other method ologies,such as CCK-8,qRT-PCR,and Western blot assays,were used to confirm improvements in cell inflammation and the survival rate by prazosin and si-Adra1a.Results Significant elevation in Adra1a protein expression in Lbp-/-primary hepatocytes was observed post-LPS stimulation(P＜0.01),whereas no notable change was found in the wildtype.A remarkable increase in the cell survival rate was noted in prazosin and si-Adra1a groups(P＜0.01,P＜0.05).Furthermore,prazosin and si-Adra1a groups exhibited significantly reduced expression of proinflammatory factors TNF-αand IL-1 β(P＜O.01),p-p38,p-ERK,and p-JNK(P＜0.01),which are associated with cell damage and inflammation.Conclusions Following LPS stimulation,upregulation of Adra1a and proinflammatory cytokine expression was observed in Lbp-/-primary hepatocytes.Specific downregulation of Adra1a expression using prazosin and si-Adra1a significantly decreased LPS-induced proinflammatory cytokines in Lbp-/-primary hepatocytes.Adra1a is implicated in the regulation of the LPS-induced inflammation response in primary hepatocytes of Lbp-/-mice.

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.

Objective To investigate the clinical features of pulmonary lymphoma and to get a better understanding of this disease. Methods Clinical data of 253 lymphoma patients in the Department of Lymphoma and Hematology in Jilin Cancer Hospital from October 2014 to March 2017 were retrospectively analyzed. The patients were divided into 30 cases of pulmonary lymphoma (lung lymphoma group) and 223 cases of non-pulmonary lymphoma (the control group). Rate assay and latex turbidimetry was used to detect lactic dehydrogenase (LDH) and β2macroglobulin (β2-MG) respectively. The expressions of programmed death 1 (PD-1), programmed death ligand 1 (PD-L1) and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) in peripheral blood CD4 +CD8 +T lymphocytes were detected by using flow cytometry. The count and measurement data of both groups were compared by using χ 2test and t test respectively. Results The patients in pulmonary lymphoma group showed secondary lesions. The proportion of smoking people in pulmonary lymphoma group was higher than that in the control group [43.3 % (13/30) vs. 24.2 % (54/223), χ 2＝ 4.964, P＝ 0.026]. The proportion of the patients in Ⅲ-Ⅳ stage in pulmonary lymphoma group was higher than that in the control group [93.3 % (28/30) vs. 57.0 % (127/223), χ2＝ 14.750, P < 0.001]. The proportion of the patients with higher international prognostic index (IPI) score in pulmonary lymphoma group was higher than that in the control group (χ2＝ 21.888, P < 0.001). The proportion of the patients with increased expression of β2-MG in pulmonary lymphoma group was higher compared with the control group [66.7 % (20/30) vs. 50.2 % (112/223), χ2＝6.682, P ＝0.091]. The proportion of the patients with the increased LDH was higher compared with the control group [63.3 % (19/30) vs. 41.5 % (86/223)], and the difference was statistically significant (χ2＝ 6.682, P ＝ 0.010). Diffuse large B-cell lymphoma (DLBCL) was the common pathological type in pulmonary lymphoma group (15 cases), followed by Hodgkin lymphoma (7 cases); imaging showed single mass or nodular type, multiple masses or nodular type, bilateral pulmonary infiltration, pleural effusion were 36.7 % (11/30), 30.0 % (9/30), 63.3 % (19/30) and 36.7 % (11/30), respectively. There were no statistical differences in the protein expression of immune check points such as PD-1, PD-L1 and CTLA-4 in both groups (all P ＞ 0.05). Conclusions Pulmonary DLBCL should be considered a secondary disease, but not a primary lesion. Smoking history is a risk factor for lymphoma patients with pulmonary involvement. Pulmonary lymphoma is similar to other extra-nodal lymphoma with high IPI scores, advanced stage and elevated LDH.

This study was aimed to explore the mechanism of baicalin on high altitude cerebral hypoxia-ischemia on mice and its influence on related target protein expressions. Morris water maze was used to screen 50 Kunming mice, which were randomly divided into the model group, control group, the low dose (0.05 mg·kg-1), middle dose (0.20 mg·kg-1) and high dose (0.60 mg·kg-1) baicalin group, with 10 rats in each group. The space memory and learning ability of mice were tested. The animal cabin with low oxygen (simulating at 4 000 m altitude) was used to establish the stable high altitude cerebral hypoxia-ischemia mouse model. Changes on SOD content, GSH-PX activities and MDA content in hippocampal tissues of mice were detected. The expressions of different target proteins, including cleaved-caspase 3, P-AKT, GFAP, Bax and Bcl-2 in brain stem of mice were detected by western blot. The results showed that the latent period of the model group was obviously longer than that of the control group (P < 0.05). The latent period of high dose baicalin group was shorter than the model group with significant difference (P< 0.05). Therefore, the best effective dose of baicalin was 0.60 mg·kg-1. Compared with the control group, the content of MDA in the hippocampal tissues of mice in the model group was significantly increased; the SOD and GSH-PX activity were obviously reduced (P < 0.05). Compared with the model group, the SOD and GSH-PX activity were obviously increased in the brain tissues of mice in the high dose baicalin group; and the content of MDA was obviously reduced (P < 0.05). From the level of protein changes, the stripes of cleaved-caspase 3, P-AKT, GFAP protein expressions in the model group were strengthened compared to the control group; the ratio of Bax/Bcl-2 was also obviously increased (P < 0.05). The expression of the baicalin group was lower than that of the model group (P < 0.05). Among them, the expression of the high dose baicalin group was the lowest. It had certain dose-response relationship. It was concluded that baicalin had protective effect on high altitude cerebral hypoxia-ischemia. Its mechanism may be related to its powerful oxidation resistance and its inhibition on expression of different target proteins, including cleaved-caspase 3, P-AKT, GFAP, Bax, Bcl-2 for the change of apoptotic pathway.

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.

This study was aimed to establish an HPLC method for simultaneous determination of the content of echinacoside, acteoside and isoacteoside in Total Cistanchis glycosides Capsules. Waters C18 column (4.6 mm ×150 mm, 5 μm) was used with the mobile phase of methanol-0.1% formic acid at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 330 nm. The column temperature was maintained at 30℃ . The results showed that the linear ranges of echinacoside, acteoside and isoacteoside were in the range of 27.792-277.92 μg·mL-1 (r = 0.9996,n = 6), 2.4184-24.184 μg·mL-1 (r = 0.9996, n = 6), 5.106-51.06 μg·mL-1 (r = 0.9998, n = 6). The average recoveries of three components were in accordance with the determination requirement. It was concluded that the method was simple and accurate, which can be used in the content determination of echinacoside, acteoside and isoacteoside in Total Cistanchis glycosides Capsules.

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 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

To investigate the modulation on the P-glycoprotein in the jejunum by combined use of Glycyrrhiza inflata and Kansui with ussing chamber and rt-pcr, Rhodamine 123 (R123), a P-gp substrate and fluorescein sodium (CF), a model drug of non-P-gp substrate transported by a passive diffusion were taken as investigational drugs. Because these two drugs can be easily assayed and widely used in various research fields. The permeability of R123 or CF via Wistar rat jejunum membranes was evaluated by in vitro ussing chamber after oral administration of four different decoctions of Glycyrrhiza inflata and Kansui for 1 week. And the concentration of R123 or CF was determined by the fluorospectrophotometry in the receiving solution. Meanwhile the expression of mdr1a in P-glycoprotein was detected by real-time fluorescent quantitative PCR. After oral administration of combined decoction of the single drug, the absorptive directed permeability of R123 increased significantly (P < 0.01). On the other hand, Kansui and combine decoction of the two drugs also decrease the permeability of secretory directed transport (P < 0.05). No action of Glycyrrhiza inflata was found on the secretory transport of R123 [Papp = (2.56 +/- 0.38) x 10(-5), cm x s(-1)] across the jejunum tissues, while Papp of control group was found [Papp = (2.35 +/- 0.27) x 10(-5), cm x s(-1)]. After oral administration of Kansui decoction for 1 week and 2 weeks, the levels of mdr1a expression in Wistar rats were lower than that of the control group, but there were no significant difference in the results. Meanwhile, Glycyrrhiza inflata had no effect on transport of CF across the jejunum tissues, though the other three groups could decrease the permeability of CF, as compared with control group. Kansui may slightly inhibit P-glycoprotein function in the intestinal membrane. For another, some compositions in Kansui inhibit P-glycoprotein function, and some others strengthen the tight junction between cells in the intestinal membrane to decrease permeability of CF. As the inhibitory action to P-glycoprotein was enhanced by combination of Glycyrrhiza inflata and Kansui, based on the results, it may be one of the mechanisms of creating toxicity once co-administration of Glycyrrhiza inflata and Kansui.