1.Thymosin ?1 increases proliferation and VEGF synthesis in human umbilical vein vessel endothelial cells in vitro
Shangkun TIAN ; Xiaohong TAO ; Hongzhou XIANG ; Yuanhu WANG
Journal of Third Military Medical University 2003;0(08):-
Objective To investigate the effects of thymosin ?1 on the proliferation of human umbilical veinssel endothelial cells(HUVECs) and synthesis of vascular endothelial growth factor(VEGF).Methods HUVECs(ECV-304) were cultured with the treatment of 100 ?l thymosin ?1 at 0,5 or 2.5 ?g/ml for 1 d in vitro.The proliferation of HUVECs were measured with MTT assay,cell cycle was tested with flow cytometry and the synthesis of VEGF was detected with Western blot analysis.Results Thymosinal promoted the proliferation of HUVECs,and the cells in S-phase was increased,and obviously promoted the synthesis of VEGF.Conclusion Thymosin ?1 promotes the proliferation of HUVECs and increases the synthesis of VEGF in HUVECs,and may have the effect of increasing the angiogenesis in the process of wound tissue.
2.Construction of lentiviral vector-mediated siRNA knockdown of ER-α36 and its action on gastric cancer cell growth
Xuming WANG ; Xuan HUANG ; Zhengqi FU ; Feng ZOU ; Shangkun ZHANG ; Zhaoyi WANG ; Lijiang LIU
Chinese Journal of Pathophysiology 2014;(12):2113-2119
[ ABSTRACT] AIM:To construct a lentiviral vector for stable delivery of the ER-α36 gene and to detect its effect on SGC7901 cell growth.METHODS: The efficient RNAi targeting sequences identified for the ER-α36 gene were screened.The Oligo DNA was synthesized with target sequences and annealed to form double-stranded DNA.Then it was digested by Xho I and EcoR I and connected with GV307 vector to produce LV-ER-α36-RNAi lentiviral vector.PCR was used to screen the positive clones and sequence.The LV-ER-α36-RNAi, pHelper 1.0 and pHelper 2.0 plasmids were co-transfected into 293T cells for producing lentiviral vector and infecting SGC7901 cell line.Fluorescence microscopy, real-time PCR and Western blotting were used to detect the transfection efficiency and gene silencing effect.17β-estrodial at concentration of 1 ×10 -10 mol/L was used to stimulate the recombinant cell line, and the action on the growth of gastric cancer cells and the expression of Src, ERK1/2 and cyclin D1 were determined.RESULTS: DNA sequencing analysis confirmed the identity of recombinant shRNA expression vectors.Immunofluorescence assay demonstrated that transfection efficiency was above 80%.Transfection of LV-ER-α36-RNAi significantly knocked down the expression of ER-α36 at mR-NA and protein levels with tetracycline ( TeT) simulating as revealed by real-time PCR and Western blotting.Compared with control group, the growth of the recombinant cell line declined and the expression of Src, ERK1/2 and cyclin D1 and the activation of Src decreased (P<0.05).CONCLUSION: Lentiviral vectors that silence ER-α36 expression are con-structed successfully and can be used to study the role of ER-α36 in gastric cancer.The ER-α36 is related with many kinds of cancer cell growth, including gastric cancer cells.
3.Effects of calcitonin gene-related peptide on aquaporin 1 and aquaporin 5 expressions in young rats with acute lung injury
Hongmin FU ; Wensha NIE ; Tingyun YUAN ; Kai YU ; Shangkun WANG ; Jiawu YANG ; Mao FAN ; Tiesong ZHANG
Chinese Journal of Applied Clinical Pediatrics 2017;32(18):1389-1392
Objective To explore the effect of calcitonin gene-related peptide (CGRP) on expressions of aquaporin (AQP)1 and AQP 5 in young rats with acute lung injury (ALI) caused by lipopolysaccharide (LPS).Methods Eighty-four young rats were randomly divided into control group,ALI model group and CGRP group.The rats in ALI model group were given intraperitoneal injection of LPS (5 mg/kg)for 2,6,12,24 hours;while the rats in CGRP group were given intraperitoneal CGRP (1 mg/kg) after 1 h injection of LPS.At 2 h,6 h,12 h and 24 h,all rats were sacrificed and lung tissues were obtained.The histopathological changes in lung tissues were evaluated by adopting hematoxylin-eosin staining,and wet/dry(W/D) was measured.The mRNA and protein levels of AQP1 and AQP5 in lung tissues were detected by adopting fluorogenic quantitative PCR (qPCR) and Western blot.Results Pathological stain showed that rats in control group had a normal lung tissue structure,and LPS made lung tissue edema,narrowing the alveolar cavity and inflammatory cell infiltration.CGRP attenuated the effect of LPS on rat's lung.The W/D ratio of lung tissue was significantly higher than that in the control group,and CGRP reduced the W/D ratio of lung tissue.qPCR showed that the mRNA levels of AQP1 and AQP5 from rats in ALI group (0.009 ±0.001 and 0.055 ±0.006)decreased compared with those in the control group (0.035±0.002 and 0.167 ±0.006) and CGRP group (0.024 ± 0.002 and 0.134 ± 0.012) (all P < 0.001).Western blot results showed after 24 h injection of LPS,both AQP1 and AQP5 levels from ALI group (0.397 ± 0.041 and 0.215 ± 0.029) were significantly lower than those in the control group (0.850 ± 0.020 and 0.741 ± 0.032) (all P < 0.001),and their levels in CGRP group (0.593-± 0.065 and 0.461 ± 0.039) were also lower than those in the control group,but higher than those in ALI group (all P < 0.001).Conclusion CGRP can enhance AQP1 and AQP5 levels and reduce pulmonary edema,and it has a protective effect on rats with acute lung injury.
4.Meta-analysis of the effect of acupuncture combined with antiemetics on prevention and treatment of nausea and vomiting after general anesthesia
Shangkun SI ; Hui LIU ; Fashuai WANG ; Weiliang ZHANG ; Dongbin ZHANG ; Fan SU
The Journal of Clinical Anesthesiology 2023;39(11):1178-1184
Objective To evaluate the effect of acupuncture(manual acupuncture,electroacu-puncture)on prevention and treatment of nausea and vomiting after general anesthesia.Methods By searching CNKI,VIP,China Biomedical Literature Database,Wanfang Medical,Embase,PubMed,Co-chrane Clinical Trial Registry,randomized controlled trial(RCT)on acupuncture combined with antiemetics for the prevention and treatment of nausea and vomiting after general anesthesia were performed.The retrieval period was from the establishment of the database to September 2022.RevMan 5.3 was used for statistical analysis.Results Fifteen studies involving 1 493 patients were included.There were 741 patients in the antiemetic group and 752 patients in the acupuncture combined antiemetic group.The incidence of postoperative nausea in the acupuncture combined with antiemetics group was significantly lower than that in the simple antiemetics group(OR=0.43,95%CI 0.34-0.54,P<0.001).The incidence of postopera-tive vomiting in the acupuncture combined with antiemetics group was significantly lower than that in the simple antiemetics group(OR=0.55,95%CI 0.45-0.66,P<0.001).Conclusion The incidence of postoperative nausea and vomiting was lower in patients treated with acupuncture(manual acupuncture,electroacupuncture)combined with antiemetics than with antiemetics alone.