Expression and localization of Copine-3-enhanced green fluorescent protein fusion protein / 解剖学报

Objective To construct the eukaryotic expression vector plasmid enhanced green fluorescent protein (pEGFP)-N1-CPNE3, and identify the expression and localization of Copine-3 protein in cells. Methods The Copine-3 coding sequences (CPNE3) was amplified by RT-PCR from human bronchial epithelial (HBE) cells and inserted into eukaryotic expression vector pEGFP-Nl. The recombinant plasmid pEGFP-Nl-CPNE3 was confirmed by endonuclease digestion and sequencing before it was transfected into 293T and H1299 cells. Cellular localization of Copine-3-EGFP fusion protein was detected by con-focal laser scanning. Expression of Copine-3 in 293T and H1299 cells was detected by Western blotting analysis. Localization of Copine-3 in clinical samples of the lung adenocarcinoma patients was detected by immunohistochemistry. Results CPNE3 was successfully constructed into the eukaryotic expression vector pEGFP-Nl and expressed in 293T and H1299 cells. Furthermore, the location of Copine-3 protein in cytoplasm and nucleus was determined by immunofluorescence staining, immuno Western blotting and immunohistochemistry in those cells and clinical samples. Conclusion The eukaryotic expression vector pEGFP-Nl-CPNE3 is constructed successfully, and Copine-3 protein is localized in cytoplasm and nucleus.

Analysis of optimal dose of thoracic radiotherapy for patients with extensive stage small cell lung cancer / 中华放射肿瘤学杂志

Objective To investigate the optimal dosage of thoracic radiotherapy in patients diagnosed with extensive stage small cell lung cancer (ES-SCLC).Methods Clinical data of ES-SCLC patients admitted to Tianjian Medical University Cancer Institute& Hospital between February 2010 and October 2015 were retrospectively analyzed.All patients received the first-line induction chemotherapy.Subsequently,216 patients without progression after the first-line induction chemotherapy were apportioned to the thoracic radiotherapy (n=180) group and chemotherapy alone group (n=36).According to the distribution characteristics of the biological equivalent dose,all patients were assigned into the A (31.3-40.2 Gy,n=23),B (46.0-46.8 Gy,n=38),C (49.5-53.7 Gy,n=43) and D groups (55.1-60.6 Gy,n=76).For the subgroup analysis,the low (31.3-46.8 Gy,n=61) and high dose groups (49.5-60.6 Gy,n=119) were divided.Kaplan-Meier survival analysis was used for prognostic analysis.Cox's regression model was conducted for multivariate prognostic analysis.Propensity score matching was utilized to control the confounding variables.Results The median overall survival of all patients was 13.2 months,and 8.3,11.0,15.8,17.8 and 8.1 months for patients in the A,B,C,D and chemotherapy alone groups,respectively (all P=0.000).The median overall survival did not significantly differ between A and B/ chemotherapy groups (P=0.172,P=0.495),and similar results were obtained between the C and D groups (P=0.624).The median overall survival in the B group was significantly longer than that in the chemotherapy alone group (P=0.020).Statistical significance was noted between C and D groups,and A and B groups (all P＜0.05).The median progression-free survival for all patients was 8.7 months,and 6.5,7.6,11.8,12.4 and 6.1 months in the A,B,C,D and chemotherapy alone groups,respectively (all P=0.000).The median progression-free survival did not significantly differ between A and B chemotherapy groups (P=0.588,P=0.668).The progression-free survival in the B group was slightly longer than that in the chemotherapy group without statistical significance (P=0.070).No statistical significance was observed between the C and D groups (P=0.627).Statistical significance was noted between C and D groups,and A and B groups (all P＜0.02).Uni-and multi-variate analyses prompted that the number of metastatic lesions and dose of thoracic radiotherapy were the independent predictors of the overall survival and progression-free survival (both P＜0.05).Concurrent chemoradiotherapy was the independent predictor of the overall survival (P=0.018).After the propensity score matching,the median overall survival and progression-free survival significantly differed between the low (n=50) and high dose groups (n=50) (10.9 vs.17.5 months,P=0.045;7.4 vs.10.7 months,P=0.014).Conclusions A relatively high dose ranging from 49.5 to 53.7 Gy is recommended during thoracic radiotherapy for ES-SCLC patients.An excessively low dose (≤ 40.2 Gy) probably fails to prolong the survival time,and an extremely high dose (≥55.1 Gy) cannot enable the patients to obtain survival benefits.

Therapeutic efficacy of three bispecific antibodies on rheumatoid arthritis mice models / 药学学报

In order to obtain the lead compound for treatment of rheumatoid arthritis (RA), in this study, therapeutic efficacy of three bispecific antibodies (BsAB-1, BsAB-2 and BsAB-3) against both hIL-1beta and hIL-17 were compared on CIA model mice. First, by ELISA method we compared the binding capacity of the three bispecific antibodies to the two antigens. The results showed that all three antibodies could simultaneously bind both antigens, among these antibodies, BsAB-1 was superior over BsAB-2 and BsAB-3. CIA model was established with chicken type II collagen (CII) and developed RA-like symptoms such as ankle swelling, skin tight, hind foot skin hyperemia. The CIA mice were treated with three antibodies once every two days for total of 29 days. Compared with the CIA model mice, the RA-like symptoms of the antibody treated-mice significantly relieved, while the BsAB-1 treated-mice were almost recovered. CII antibody level in the serum and cytokines (IL-2, IL-1beta, IL-17A and TNF-alpha) expression in the spleen were examined. Compared with the CIA model mice, all three antibodies could significantly reduce CII antibody and cytokine expression levels. BsAB-1 antibody was more potent than BsAB-2 and BsAB-3. In summary, BsAB-1 is superior over BsAB-2 and BsAB-3 in amelioration of RA symptoms and regulation of CII antibody production and pro-inflammatory cytokine expression, therefore, BsAB-1 can be chosen as a lead compound for further development of drug candidate for treatment of RA.

Expression of serum cytokines in patients with pneumoconiosis by antibody chip / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the changes in expression of serum cytokines in patients with pneumoconiosis using cytokine antibody chips (CACs).</p><p><b>METHODS</b>The CAC technology was applied to measure the serum levels of 60 cytokines in 12 patients with pneumoconiosis and 3 normal controls.</p><p><b>RESULTS</b>In the patients with pneumoconiosis, the highly expressed serum cytokines included interleukin (IL)-1α, IL-1β, IL-2, ILs 4-16, macrophage colony-stimulating factor, interferon-γ, tumor necrosis factor (TNF)-α, TNF-β, human bone morphogenetic protein-6, fibroblast growth factor-7, neurotrophin-3, and stem cell factor, and the lowly expressed serum cytokines included recombinant human I-309, monocyte chemoattractant protein (MCP)-1, MCP-2, MCP-3, MCP-4, macrophage inflammatory protein (MIP)-1-δ, and MIP-3-α.</p><p><b>CONCLUSION</b>Patients with pneumoconiosis have changes in the expression of most serum cytokines.</p>

Prokaryotic expression and immunogenicity analysis of glycoprotein from infectious hematopoietic necrosis virus / 病毒学报

In order to detect Infectious hematopoietic necrosis virus with immunological methods, the surface glycoprotein of a recent IHNV-Sn isolated from farmed rainbow trout ( Oncorhynchus mykiss ) in China was amplified and cloned into pET27b(+) vector (designated as pET27b-G ). The expression of recombinant plasmid pET27b-G in E. coli BL21(DE3) was induced and determined by SDS-PAGE analysis. The predicted molecular weight of glycoprotein protein was approximately 55 kD and was confirmed in this study. The inclusion body of glycoprotein was treated with urea at different urea concentrations, and dialyzed into PBS buffer. Purified glycoprotein with high concentration was obtained after dialyzed in the PBS buffer. Antisera against glycoprotein were produced from immunized rabbits. The prepared antisera could react specifically with both the recombinant glycoprotein and natural glycoprotein of the IHNV-Sn isolated in the test of indirect ELISA, and the titer against the recombinant glycoprotein was 1:20,000. IFA showed that the antisera can recognize the glycoprotein located on the surface of IHNV-Sn and IHNV reference strain. These results indicated that the expressed glycoprotein was immunogenical and antigenical and could be functional as the natural IHNV glycoprotein. These results established a foundation for further study on vaccine and rapid diagnosis of IHNV.

Expression of GADD153 in follicular tumors of thyroid and comparison with CK19, Galectin-3 and HBME-1 / 中华病理学杂志

<p><b>OBJECTIVE</b>To study immunohistochemical expression of GADD153 and assess its usefulness as markers in the differential diagnoses in follicular tumors of the thyroid.</p><p><b>METHODS</b>Immunohistochemical staining was performed on formalin-fixed paraffin-embedded tissue of 34 cases of follicular thyroid adenomas (FTA), 46 cases of follicular thyroid carcinomas (FTC), 29 cases of follicular variant papillary carcinomas (FVPC).</p><p><b>RESULTS</b>(1) GADD153 was expressed in cell nucleus with positive or strong positive expression in FTC, and no or weak expression in FTA and FVPC. The positive expressions of GADD153 were present in 38 of 46(82.6%) in FTC, 11 of 34(32.4%) in FTA and three of 29(10.3%) in FVPC, the positive expression rate in FTC was obviously higher than that in FTA and in FVPC, the differences were statistically significant (χ² = 20.80 and 37.48; P < 0.01). (2) CK19, Galectin-3 (Gal-3) and HBME-1 were all expressed in the cytoplasm, the positive expressions of CK19, Gal-3 and HBME-1 were present in 54.3% (25/46), 67.4% (31/46) and 58.7% (27/46) in FTC; 50.0% (17/34), 29.4% (10/34) and 32.4% (11/34) in FTA; 100% (29/29), 93.1% (27/29) and 89.7% (26/29) in FVPC, the differences were statistically significant as well (χ² = 21.20 and 8.22; P < 0.01). (3) According to the expressions of CK19, Gal-3, HBME-1 and GADD153, we divided the results into low expression group (0 or 1+) and high expression group (2+ or 3+), the sensitivity and the specificity were calculated. in FTA, the sensitivity were 26.5%, 8.8%, 2.9% and 11.8%; the specificity were 50.7%, 52.0%, 54.7% and 58.7%. in FTC, the sensitivity were 19.6%, 26.1%, 23.9% and 65.2%; the specificity were 41.3%, 57.1%, 62.0% and 92.1%. in FVPC, the sensitivity were 96.6%, 82.8%, 79.3% and 3.4%; the specificity were 77.5%, 81.3%, 85.0% and 57.5%.</p><p><b>CONCLUSIONS</b>The sensitivity and the specificity of GADD153 expression are well for diagnosing FTC, and CK19, Gal-3, HBME-1 are well for FVPC. The four markers when used in combination, are better to identify the follicular tumors of the thyroid.</p>

Clinical study on multi-segmental pedicle screw implantation in the treatment of middle-upper thoracic spine fracture and dislocation / 中国骨伤

<p><b>OBJECTIVE</b>To review a group of clinical data of pedicle screw implantation in the treatment of middle-upper thoracic fracture and dislocation and explore these clinical effect.</p><p><b>METHODS</b>Through the retrospective analysis of 23 patients (31 vertebraes) of middle-upper thoracic fracture and dislocation included 21 male and 2 female,aged from 20 to 47 years in average of 33.5 years. The location and type of fractures were as follows: 2 in T2, 2 in T3, 5 in T4, 6 in T5, 10 in T6, 4 in T7, 2 in T8. Three cases were compression fractures, 7 burst fractures, 9 fracture-dislocations, 4 burst-dislocations.</p><p><b>RESULTS</b>After a followed-up from 3 to 48 months, average 25.5 months, all cases had no serious complications. Anterior height of affected vertebral bodies had been restored from 40.4% to 90.3% after operation. Patients with incomplete spinal cord injuries improved one to two grades according to ASIA grade of spinal cord injury. Patients with complete spinal cord injuries improved 21.7 scores on average according to Sensory and Motion Scores.</p><p><b>CONCLUSION</b>Multi-segmental pedicle screw implantation for the injuries of vertebral body may get satisfactory reduction and fixation effects, and prevent the long-term loss of the vertebral body height and implanting failure.</p>

Effect of in vitro and in vivo treatment with mitomycin C on activities of CYP2D1/2, CYP2C11, and CYP1A2 in rat liver / 药学学报

<p><b>AIM</b>To evaluate the effect of in vitro and in vivo treatment with mitomycin C (MMC) on activities of CYP2D1/2, CYP2C1 , and CYP1A2 in the liver of male rats.</p><p><b>METHODS</b>Using HPLC to determine the activities of the three isoenzymes in rat liver microsomes by detecting the specific metabolites of their substrates after treatment with inducers in vivo or inhibitors in vitro.</p><p><b>RESULTS</b>In vitro, MMC inhibited the activity of CYP2D1/2, CYP2C11, and CYP1A2 in dexamethasone-induced microsomes by (19 +/- 6)% (P < 0.05), (85 +/- 10)% (P < 0.01), and (36 +/- 6)% (P < 0.05), respectively, and decreased the activity of CYP1A2 in beta-naphthoflavone-induced microsomes by (58 +/- 6)% (P < 0.01). Rats were injected intraperitoneally with 20% of the LD50 of MMC for 3 or 6 d. The treatment showed no significant effect on microsomal activities of CYP2D1/2, CYP2C11 or CYP1A2.</p><p><b>CONCLUSION</b>MMC can inhibit the activities of CYP2D1/2, CYP2C11, and CYP1A2 in rat liver microsomes in vitro, but it showed no significant effect on the activities of the three isoenzymes in vivo.</p>