1.Eeffect of K65R site-mutagenesis of cataracts-related gene Hsf4b on downstream heat shock proteins expression
Qiang LOU ; Panpan XIE ; Xiukun CUI ; Yuanfang MA ; Yanzhong HU
Chinese Journal of Pathophysiology 2015;(9):1699-1703
AIM:To clarify the impact of heart shock factor 4b (Hsf4b) K65R mutation on the regulation of downstream protein expression .METHODS:Non-functional Lys mutant plasmid pWZL-blast-HA-Hsf4b/K65R was genera-ted by replacing single , homologous amino acids using KOD-Plus-Mutagenesis-Kit.Mouse lens epithelial mLEC stable cell lines expressing Hsf4b or Hsf4b/K65R were constructed by lentivirus infection .The expression of Hsf4b in the mutant and the wildtype mLEC cells was confirmed by Western blotting .The expression of Hsf4b downstream proteins such as heat shock protein ( Hsp)70, Hsp90, Hsp27 and CryAB was examined by Western blotting and real-time PCR.RESULTS:The results of PCR and DNA sequencing confirmed the successful construction of mLEC Hsf 4b/K65R mutant.The K65R mutation didn’t influence Hsf4b expression in the mLEC cells.After K65R mutation in Hsf4b, the expression levels of Hsp27 and CryAB were down-regulated and the expression of Hsp 70i and Hsp90a upregulated.CONCLUSION: pWZL-blast-HA-Hsf4b/K65R can be used to construct a stable cell line by infecting with lentivirus .Hsf4b/K65R mutation influ-ences the regulation of downstream heat shock proteins .
2.Endoscopic palliative treatment for advanced pancreatic cancer
Shuren MA ; Zhuo YANG ; Ning ZHANG ; Xiukun ZONG ; Xuefeng ZHANG ; Huiyong JIANG ; Fei GAO ; Zhifeng ZHAO ; Yunfeng ZHAO ; Feng GAO
Chinese Journal of Digestive Endoscopy 2011;28(8):452-455
Objective To investigate the therapeutic effects of endoscopy for palliative treatment of advanced pancreatic cancer. Methods A typical case of un-resectable advanced pancreatic cancer was reviewed, who underwent obstruction of upper gastrointestinal tract, obstructive jaundice and alimentary tract hemorrhage subsequently. The patient received multiple placement of intestinal tract stents, common bile duct stents and hemostatic treatment endoscopically. Because of the obstruction of upper gastrointestinal tract, jejunalostomy and retrograde endoscopy through the orificium fistulae were performed to place bile duct stents. Results The patient survived for 10 months with good life quality after diagnosis, obstruction of upper gastrointestinal tract, obstructive jaundice and alimentary tract hemorrhage were cured and didn't recur till death.Conclusion Therapeutic endoscopy, safe and effective, is the first choice for advanced pancreatic cancer complicated with obstruction of digestive tract (including gastrointestinal tract, bile duct and pancreatic duct).
3.Quality effect of systemic psychological intervention for patients with craniofacial trauma before MRI scanning
Xiukun WANG ; Pengfei ZHAO ; Junxia BAI ; Tong GAO ; Xiaoqing MA
Chinese Journal of Modern Nursing 2015;(15):1783-1785
Objective To investigate the psychological influence of anxiety and depression of patients with craniofacial trauma using psychological intervention before MRI scanning. Methods One hundred and forty hospitalized patients with craniofacial trauma were enrolled averagely divided into psychological experimental group and control group by time sequence. The patients of the experimental group received psychological intervention before MRI scanning, whereas the patients of the control group did not, but patients in both groups underwent routine nursing. The psychological state of all the patients were evaluated using self-rating anxiety scale ( SAS) and self-rating depression scale ( SDS) . Results There was no difference in either SAS or SDS between two groups before intervention (P>0. 05). After intervention, the anxiety score and depression score were (38. 96 ± 10. 15) and (46. 86 ± 11. 25), which were lower than (43. 53 ± 9. 32) and (52. 51 ± 6. 80) of the control group (t= -2. 776, -3. 601, respectively;P <0. 05). All patients in the experimental group completed examination, but two cases failed in the control group. The image quality control in the experimental group were excellent ( clear and no constructed defect ) , and there were three cases of good image for quality control in the control group. Conclusions Psychological intervention before MRI scanning may play an important role in improving the psychological state of patients with craniofacial trauma so as to improve the cooperation of examination, completion rate and quality of image.
4.Effects of miR-17-5p targeting B7-H3 on biological characteristics and immunomodulatory effects of colorectal cancer cells
Yang HUA ; Xiukun MA ; Zhu HONG ; Xianglong LIU
Chinese Journal of Immunology 2024;40(7):1431-1435,1440
Objective:To investigate the regulatory role of miR-17-5p on B7-H3 and its immunomodulatory role in the develop-ment of colorectal cancer.Methods:Twenty-five pairs of human colorectal cancer tissues and adjacent normal tissues were collected,and the expression levels of miR-17-5p,B7-H3 and PD-L1 were analyzed by real-time fluorescence quantitative PCR,protein immuno-blotting and immunohistochemical assays.Human colorectal cancer HCT-116 cells were divided into four groups:miR-NC group,miR-17-5p mimic group,si-NC group and si-B7-H3 group.The growth,invasive ability and apoptosis rate of HCT-116 cells were ana-lyzed by cell colony formation assay,Transwell assay and flow cytometry.The regulatory effect of miR-17-5p on B7-H3 was analyzed by luciferase reporter gene assay.The levels of IL-2,IL-4,IL-6,IL-10,IFN-γ and TNF-α cytokines in the cell culture medium were analyzed by ELISA.Results:Compared with paracancerous tissues,miR-17-5p expression was decreased and B7-H3 expression was increased in colorectal cancer tissues.miR-17-5p mimic and si-B7-H3 were able to reduce the number of HCT-116 cell colony forma-tion and invasion,promote HCT-116 cell apoptosis,and decrease the levels of IL-2,IL-4,IL-6,IL-10,IFN-γ and TNF-α cytokines in culture medium.In addition,miR-17-5p mimic and si B7-H3 were able to inhibit PD-L1 expression in HCT-116 cells.Conclusion:miR-17-5p can targeting inhibit B7-H3 expression,affect the immunomodulatory role in colorectal cancer development,and inhibit metastasis and promote apoptosis of colorectal cancer cells.
5.Evolution and development of potent monobactam sulfonate candidate IMBZ18g as a dual inhibitor against MDR Gram-negative bacteria producing ESBLs.
Zhiwen LI ; Zhihao GUO ; Xi LU ; Xican MA ; Xiukun WANG ; Rui ZHANG ; Xinxin HU ; Yanxiang WANG ; Jing PANG ; Tianyun FAN ; Yonghua LIU ; Sheng TANG ; Haigen FU ; Jingpu ZHANG ; Yinghong LI ; Xuefu YOU ; Danqing SONG
Acta Pharmaceutica Sinica B 2023;13(7):3067-3079
A series of new monobactam sulfonates is continuously synthesized and evaluated for their antimicrobial efficacies against Gram-negative bacteria. Compound 33a (IMBZ18G) is highly effective in vitro and in vivo against clinically intractable multi-drug-resistant (MDR) Gram-negative strains, with a highly druglike nature. The checkerboard assay reveals its significant synergistic effect with β-lactamase inhibitor avibactam, and the MIC values against MDR enterobacteria were reduced up to 4-512 folds. X-ray co-crystal and chemoproteomic assays indicate that the anti-MDR bacteria effect of 33a results from the dual inhibition of the common PBP3 and some class A and C β-lactamases. Accordingly, preclinical studies of 33a alone and 33a‒avibactam combination as potential innovative candidates are actively going on, in the treatment of β-lactamase-producing MDR Gram-negative bacterial infections.