Identification and expression analysis of NHX gene family in Chinese cabbage / 生物工程学报

Na+/H+ antiporter (NHX) gene subfamily plays an important role in plant response to salt stress. In this study, we identified the NHX gene family members of Chinese cabbage and analyzed the expression patterns of BrNHXs gene in response to abiotic stresses such as high temperature, low temperature, drought and salt stress. The results showed that there were 9 members of the NHX gene family in Chinese cabbage, which were distributed on 6 chromosomes respectively. The number of amino acids was 513-1 154 aa, the relative molecular weight was 56 804.22-127 856.66 kDa, the isoelectric point was 5.35-7.68. Members of BrNHX gene family mainly existed in vacuoles, the gene structure is complete, and the number of exons is 11-22. The secondary structures of the proteins encoded by the NHX gene family in Chinese cabbage had alpha helix, beta turn and random coil, and the alpha helix occurred more frequently. Quantitative real-time PCR (qRT-PCR) analysis showed that the gene family members had different responses to high temperature, low temperature, drought and salt stress, and their expression levels differed significantly in different time periods. BrNHX02 and BrNHX09 had the most significant responses to these four stresses, and their expression levels were significantly up-regulated at 72 h after treatments, which could be used as candidate genes to further verify their functions.

Construction of home-based cardiac rehabilitation intervention system for patients after percutaneous coronary intervention / 中国实用护理杂志

Objective:To construct a home-based cardiac rehabilitation intervention system for patients after percutaneous coronary intervention, and to provide reference for improving the self-management ability and family support of home-based cardiac rehabilitation of patients after PCI.Methods:Based on the literature study and group discussions, a draft of home-based cardiac rehabilitation intervention system for patients after PCI based on empowerment theory was constructed. From January to April 2021, the Delphi method was used to conduct 2 rounds of expert consultations among 18 experts from 9 hospitals, and the items were modified according to the experts′ advice.Results:The expert positive coefficients of the 2 rounds were 94.44% and 100.00%, the expert authority coefficients was 0.91, and the Kendall coefficients were 0.188 and 0.255. Finally, a home-based cardiac rehabilitation intervention system for patients after PCI was formed, including 5 first-level items, 19 second-level items and 21 third-level items.Conclusion:The home-based cardiac rehabilitation intervention system for patients after PCI is reliable, scientificity and practical, and has guiding significance for promoting the development of home-based cardiac rehabilitation for PCI patients.

knocking out mediated by CRISPR-Cas9 genome editing for PD-L1 attenuation and enhanced antitumor immunity

Blocking the programmed death-ligand 1 (PD-L1) on tumor cells with monoclonal antibody therapy has emerged as powerful weapon in cancer immunotherapy. However, only a minority of patients presented immune responses in clinical trials. To develop an alternative treatment method based on immune checkpoint blockade, we designed a novel and efficient CRISPR-Cas9 genome editing system delivered by cationic copolymer aPBAE to downregulate PD-L1 expression on tumor cells specifically knocking out Cyclin-dependent kinase 5 () gene . The expression of PD-L1 on tumor cells was significantly attenuated by knocking out , leading to effective tumor growth inhibition in murine melanoma and lung metastasis suppression in triple-negative breast cancer. Importantly, we demonstrated that aPBAE/Cas9-Cdk5 treatment elicited strong T cell-mediated immune responses in tumor microenvironment that the population of CD8 T cells was significantly increased while regulatory T cells (Tregs) was decreased. It may be the first case to exhibit direct PD-L1 downregulation CRISPR-Cas9 genome editing technology for cancer therapy. It will provide promising strategy for preclinical antitumor treatment through the combination of nanotechnology and genome engineering.

The correlation of IL-8 signaling pathway and EGFR pathway in MDA-231 cells of breast carcinoma / 中华微生物学和免疫学杂志

Objective To study the effect of IL-8 on cell proliferation and invasion,and to analyze the correlations between chemokine and epidermal growth factor receptor(EGFR)signaling pathways in breast carcinoma cells.Methods IL-8 secretion responded to treatment with rhEGF and anti-EGFR and expression of its receptors CXCR1,CXCR2 in MDA-231 cells were measured by ELISA and immunocytochemistry,respectively.Effect of rhIL-8 and neutralizing antibody on cell proliferation and invasion were analyzed by using MTT and matrigel invasion assay.EGFR transactivation stimulated with rhIL-8 and neutralizing an tibody was assessed by Western blot using anti-phosphotyrosine antibody.Results MDA-231 cells released hish level of IL-8 and two receptom of IL-8(CXCR1 and CXCR2)both expressed on cell membrane.Exogenous IL-8 and its neutralizing antibody did not significantly influence the proliferation of breast carcinoma cells,but rhIL-8 stimulated invasive activity in MDA-231 cells and its neutralizing antibody inhibited the in vasive activity(P＜0.05).EGF and anti-EGFR both inhibited the secretion of IL-8 in breast carcinoma cells,and IL-8 had no effect on EGFR phosphorylation,but anti-IL-8 induced transactivation of EGFR after 24h.Conclusion IL-8 contributes to tumor progression in breast carcinoma through its enhancement of in vasive activitv but not act as an autocrine growth factor.The correlation of competitive inhibition rather than cross-talk is found between G protein coupled receptor(GPCR)-mediated IL-8 signaling pathway and EGFR pathway in breast carcinoma.

Protein-chip for autoantibodies profile detection / 生物工程学报

We selected 12 antigens corresponding to commonly used autoantibodies in clinical practice to prepare antigen microarray. We chose NBT/BCIP color reaction as the end detection strategy to develop a new autoantibody protein chip detection system. Using this system, we optimized the best spotting solution, spotting concentration of the 12 antigens and the dilution of serum. We prepared a protein chip that could detect 12 autoantibodies simultaneously using the optimized antigen concentration. We established a new method to determine the cutoff of each autoantibodies by evaluation of 678 positive and 120 negative serum of clinical sample. We also evaluated the sensitivity and specificity of our new detection system. The optimal spotting solution was 0.1% TBST, the dilution of serum was 1:4 and the best spotting concentration of the 12 antigens were ANA 520 microg/mL, Ro-60/SSa 465 microg/mL, La/SSb 530 microg/mL, Jo-1 530 microg/mL, Scl-70 525 microg/mL, Sm 520 microg/mL, Ro-52/SSa 615 microg/mL, RF 340 microg/mL, CCP 465 microg/mL, ulRNP 410 microg/mL, CENP-B 490 microg/mL and dsDNA 580 microg/mL respectively. It had higher coincidence rate compared to current clinical used methods. We have developed a 12 antigens protein chip for the detection of autoantibodies based on the NBT/BCIP color reaction system. This system was fast, convenient, efficient, and cost-effective.

Predication of secondary structures and epitopes of fusion protein pp150/MDBP / 生物医学工程学杂志

The secondary structures of fusion protein pp150/MDBP, including alpha-helix, beta-sheet, turn regions, were analyzed by Garnier-Robson's and Chou-Fasman's methods; the antigenic epitopes of B cells were analysed by using hydrophilicity plot. The results showed that the fusion protein pp150/MDBP might have less alpha-helix, but be rich in beta-sheet and turn regions. The epitopes recognized by B cells may be at 7-56 amino acid residues or adjacent to 137-192 amino acid residues.

Brain-type natriureric peptide and cardiovascular diseases / 基础医学与临床

It is reported by current investigations,that BNP can suppress heart hypertrophy but it can also depress compensatory hypertrophic response.BNP is an useful marker in the early diagnosis,prognosis,therapy guiding of heart failure and coronary heart disease,but comorbid conditions should be considered in the diagnosis of heart failure.The polymorphism in the 5'-flanking region of the NPPB gene is found to be related with essential hypertension.On the basis of some investigation,it is supposed that the instillation of BNP at the early stage of hypertension will contribute to therapy of hypertension.

Nonsteroidal anti-inflammatory drugs and the risk of polyposis, colon carcinoma and rectal carcinoma / 中华预防医学杂志

<p><b>OBJECTIVE</b>To probe the risk of colorectal polyp, colon and rectal carcinoma and the intake of NSAIDs.</p><p><b>METHODS</b>Case-control study participants were from patients who underwent colonoscopy at different hospitals, the persons with the above disease was as cases, and those without the above diseases was as controls. Use of NSAIDs was assessed by interviewing the participants with a questionnaire which include a list of NSAIDs and related dietary and life style factors and family history.</p><p><b>RESULTS</b>There are 37 cases of colorectal polyp, 105 cases of colon carcinoma and 142 cases of rectal carcinoma and 66 controls. Adjusted for potential confounders, the risk of colorectal polyposis, colon carcinoma and rectal carcinoma were markedly reduced by NSAIDs. The OR values were 0.21 (95% CI 0.07-0.65, P = 0.007), 0.13 (95% CI 0.05-0.35, P < 0.001), 0.15 (95% CI 0.11-0.58, P < 0.001) respectively. The risk of the above diseases were also reduced markedly by aspirin, the OR values were 0.265 (95% CI 0.07-0.96, P = 0.044), 0.10 (95% CI 0.03-0.35, P < 0.001), 0.15 (95% CI 0.04-0.49, P = 0.002) respectively. The risk of colon carcinoma was also reduced by profen, with the OR being 0.11 (95% CI 0.02-0.64, P = 0.014).</p><p><b>CONCLUSIONS</b>Aspirin and other NSAIDs could reduced the risk of colorectal polyp, colon carcinoma and rectal carcinoma markedly. Aspirin was the most prospective chemopreventive agents for colorectal polyp, colon and rectal carcinoma for its capability of reducing the risk of cardio-cerebral vascular disease as well.</p>

Preparation conditions optimization and preliminary application of the microarray for detecting autoantibodies / 中华风湿病学杂志

Objective To optimize several key factors in preparing protein microarray,and establish microarrays to detect autoantibodies.Methods Four factors,including incubation time of the microarray after spotting,the blocking reagent,the length of blocking time,the length of time conjugating with the second-an- tibody,were selected to carry out orthogonal optimization,then we determined the optimal spotting concentra- tion of antigen and the best dilution of serum with the optimized conditions obtained from the above.On the basis of the optimized conditions,we prepared microarray to detect six autoantibodies simultaneously,which were compared with IIF and ELISA.Results The best conditions we determined from the experiment were: incubating the spot microarray for 120 min,PBST containing 1% BSA and 2.5% saccharose as the blocking reagent and blocking for 30 min,reacting with the second-antibody for 30 min,the appropriate spotting con- centrations of six antigens differed from 100?g/ml to 300?g/ml and the best serum dilution was 1:2.Com- pared with IIF for ANA detection,the sensitivity of the microarray was 88.6%,the specificity was 83.3%,and compared with ELISA in detecting the other five autoantibodies,the sensitivity of the microarray was between 80.0% and 88.2%,and the specificity was between 90.2% and 98.6%.Conclusion The optimization condi- tions we obtained are suitable for preparing protein microarray,and the detection sensitivity and specificity of autoantibodies by microarray are consistent with the conventional methods in general.The microarray for de- tecting autoantibodies is applicable in the clinical diagnosis of autoimmune disease.

Inhibitory effects of piroxicam on the transplanted sarcoma S180 of mice and its effect on the expression of COX-2,VEGF, FGF-2 and MVD / 中国药理学通报

AIM To investigate the effects of piroxicam on transplanted Sarcoma S180 and the expression of COX 2,VEGF,FGF 2 and microvessel density (MVD) in tumor tissue METHODS Kunming mice were randomizedly divided into control group, FT207 positive group and 5, 2 5, 1 mg?kg -1 piroxicam groups One day after inoculation of 0 2 ml S180 cell suspension, FT207 and piroxicam were given by gastric intubation for 9 days The inhibitory rate on S180 was calculated routinely The expression of COX 2,VEGF,FGF 2 and MVD was detected by immunohistochemistry RESULTS The growth of S180 was significantly inhibited by piroxicam at the doses of 5, 2 5, 1 mg?kg -1 with the inhibitory rate of 31 4%,40 7% and 34 9% respectively The expression of COX 2 in the tumor tissue was also inhibited by piroxicam. Accordingly the expression of VEGF,FGF 2 and MVD was markedly inhibited in dose dependent manner by piroxicam CONCLUSIONS The results suggest that piroxicam has inhibitory effects on S180,and it also decreases the expression of COX 2 in tumor tissue. There is a relation ship between the expression of COX 2 and angiogenesis related factor Antiangiogenesis may be another mechanism for piroxicam to exert its chemopreventive and treatment effects.