1.Relationship between Wnt/β-catenin signaling pathway and autophagy during hyperoxia-induced acute lung injury in infantile rats
Jine JIA ; Jianbo WANG ; Zhiqiang YU
Chinese Journal of Anesthesiology 2023;43(10):1232-1236
Objective:To evaluate the relationship between Wnt/β-catenin signaling pathway and autophagy during hyperoxia-induced acute lung injury in infantile rats.Methods:A total of 24 clean-stage healthy male infantile Sprague-Dawley rats, aged 14 days, weighing 40-50 g, were allocated into 4 groups ( n=6 each) using a random number table method: control group (C group), hyperoxic acute lung injury (HALI) group, HALI+ IWP-2 group (HI group) and HALI+ DMF group (HD group). HALI model was developed by inhaling oxygen at a concentration greater than 90% for 72 h. Starting from 30 min before developing the model, IWP-2 15 mg/kg was intraperitoneally injected every day for 3 consecutive days in HI group, the equal volume of DMF solution was injected every day for 3 consecutive days in HD group, and the equal volume of normal saline was intraperitoneally injected instead in C and HALI groups. Blood samples were taken from the common carotid artery for blood gas analysis at the end of developing the model, and oxygenation index (OI) was calculated. Then the infantile rats were sacrificed under deep anesthesia, and lungs were removed for examination of the pathological changes which were scored and for determination of the weight to dry weight ratio (W/D ratio), contents of interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-alpha (TNF-α) (by enzyme-linked immunosorbent assay) and expression of Wnt3a, β-catenin, microtubule-associated protein 1 light chain 3(LC3), Beclin1 and p62 (by Western blot). LC3Ⅱ/LC3Ⅰ ratio was calculated. Results:Compared with C group, the OI was significantly decreased, the W/D ratio, lung injury score and contents of IL-6, IL-1β and TNF-α were increased, and the expression of Wnt3a, β-catenin and Beclin1 was up-regulated, the expression of p62 was down-regulated, and LC3Ⅱ/LC3Ⅰ ratio was increased in HALI, HD and HI groups ( P<0.05). Compared with HALI group, the OI was significantly decreased, the W/D ratio, lung injury score and contents of IL-6, IL-1β and TNF-α were increased, and the expression of Wnt3a, β-catenin and p62 was down-regulated, the expression of Beclin1 was up-regulated, and LC3Ⅱ/LC3Ⅰ ratio was increased in HI group ( P<0.05), and no significant change was found in the parameters mentioned above in HD group ( P>0.05). Compared with HI group, the OI was significantly increased, the W/D ratio, lung injury score and contents of IL-6, IL-1β and TNF-α were decreased, and the expression of Wnt3a, β-catenin and p62 was up-regulated, the expression of Beclin1 was down-regulated, and LC3Ⅱ/LC3Ⅰ ratio was decreased in HD group ( P<0.05). Conclusions:In the pathophysiology of hyperoxia-induced acute lung injury in infantile rats, Wnt/β-catenin signaling pathway may be a negative regulator of autophagy. Wnt/β-catenin signaling pathway may be involved in the process of HALI through negative regulation of autophagy.
2.Transcriptome and proteome analysis of pregnancy and postpartum anoestrus ovaries in yak
Zhou CHEN ; Jine WANG ; Junyuan MA ; Shuyuan LI ; Shengdong HUO ; Yanmei YANG ; Yingpai ZHAXI ; Yongqing ZHAO ; Derong ZHANG
Journal of Veterinary Science 2022;23(1):e3-
Background:
Domestic yaks are the most important livestock species on the Qinghai-Tibetan Plateau. Adult female yaks normally breed in the warm season (July to September) and enter anestrous in the cold season (November to April). Nevertheless, it is unclear how ovarian activity is regulated at the molecular level.
Objectives:
The peculiarities of yak reproduction were assessed to explore the molecular mechanism of postpartum anestrus ovaries in yaks after pregnancy and parturition.
Methods:
Sixty female yaks with calves were observed under natural grazing in Haiyan County, Qinghai Province. Three yak ovaries in pregnancy and postpartum anestrus were collected. RNA sequencing and quantitative proteomics were employed to analyze the pregnancy and postpartum ovaries after hypothermia to identify the genes and proteins related to the postpartum ovarian cycle.
Results:
The results revealed 841 differentially expressed genes during the postpartum hypoestrus cycle; 347 were up-regulated and 494 genes were down-regulated. Fifty-seven differential proteins were screened: 38 were up-regulated and 19 were down-regulated. The differential genes and proteins were related to the yak reproduction process, rhythm process, progesterone-mediated oocyte maturation, PI3K/AKT signaling pathway, and MAPK signaling pathway categories.
Conclusions
Transcriptome and proteomic sequencing approaches were used to investigate postpartum anestrus and pregnancy ovaries in yaks. The results confirmed that BHLHE40, SF1IX1, FBPX1, HSPCA, LHCGR, BMP15, and ET-1R could affect postpartum hypoestrus and control the state of estrus.
3.Schisandrin B induces apoptosis of human breast cancer MDA-MB-231 cells through ROS mediated endoplasmic reticulum stress
Weiting WANG ; Xueqin YIN ; Jine XIA ; Xiayan ZHANG
Journal of Pharmaceutical Practice 2021;39(6):499-503
Objective To study the effects of schisandrin B (Sch B) on the apoptosis of human breast cancer MDA-MB-231 cells and its mechanism. Methods Cell counting reagent (CCK-8) was used to detect the effect of Sch B on the survival rate of MDA-MB-231 cells. MDA-MB-231 cells were treated with Sch B (10, 20, 40 μmol/L) for 24 hours. The cell death was detected by Annexin V-FITC/PI. The levels of intracellular reactive oxygen species (ROS) were detected by DCFA-DA fluorescent probe. Apoptosis and the expression of endoplasmic reticulum stress related proteins (Bcl-2、Bax、CHOP、GPR78、PERK、p-PERK、p-eIF2α、eIF2) were detected by Western blot. Results Compared with the blank group, the cell survival rate decreased significantly (P<0.01) with the increase of Sch B concentration, and its IC50 was 19.16 μmol/L. Compared with the control group, Sch B groups (10, 20, 40 μmol/L) inhibited cell clone formation in a dose-dependent manner (P<0.05). Sch B groups (10, 20, 40 μmol/L) induced apoptosis (P<0.05), significantly reduced the expression of anti-apoptotic protein Bcl-2 and significantly increased the expression of pro-apoptotic protein Bax (P<0.05). Sch B groups (10, 20, 40 μmol/L) significantly increased the level of intracellular ROS in a dose-dependent manner (P<0.05). Sch B groups (10, 20, 40 μmol/L) stimulated endoplasmic reticulum stress and increased the expressions of endoplasmic reticulum stress-related proteins CHOP, GPR78 and p-eIF2α in a dose-dependent manner (P<0.05). Conclusion Sch B induces apoptosis of MDA-MB-231 cells through ROS mediated endoplasmic reticulum stress.
4.High-resolution 3.0T magnetic resonance imaging in diagnosis of ligament and tendon injuries of the forefoot
Yaxiong LI ; Heng ZHANG ; Rongjie BAI ; Jine WANG ; Yupeng CAO ; Huili ZHAN ; Zhanhua QIAN
Chinese Journal of General Practitioners 2021;20(6):676-681
Objective:To evaluate the application high-resolution 3.0-T magnetic resonance imaging (MRI) in diagnosis of ligament and tendon injuries of the forefeet.Methods:From September 2014 to August 2020, 40 patients with forefeet injury (40 forefoot) and 20 healthy subjects (40 forefoot) meeting the including and excluding criteria were enrolled into this retrospective study. All subjects underwent MRI T1-weighted image (T1WI) and fat suppression proton density-weighted imaging (PD-FS) scan on coronal planes, sagittal planes and axial planes, respectively. Then the MRI characteristics of tendons in forefoot of healthy subjects and patients with forefoot injuries were compared. In patients the MR features of ligament and tendons injury were compared with the surgical findings or follow-up results.Results:In 20 normal subjects the tendons and ligament of the forefoot showed homogeneous low-signal-intensity on T1WI and PD-FS images. In the 40 patients with forefoot injuries, 22 cases were confirmed by surgical findings and 18 cases were confirmed by follow-up results. There were 8 cases of collateral ligament injuries, including 2 cases with complete tear, 3 cases with partial tear and 3 cases with sprain; 2 cases of extensor tendon with partial tear; 8 cases of flexor tendon injuries, including 3 cases with complete tear, 3 cases with partial tear and 2 cases with sprain; 3 cases of abductor tendon with partial tear; 13 cases of plantar plate injuries; and 6 cases of Lisfranc ligament injuries including 5 cases with complete tear and 1 case with partial tear. The injured ligament and tendons presented thicker and discontinuity in T1WI; and demonstrated heterogeneously increased signal intensity with edema in the surrounding soft tissues in PD-FS. The partially torn ligament and tendons showed discontinuity and thickening; and the completely torn ligament and tendons showed that they were interrupted and retracted, fiber structure disappeared and was filled with liquid, and edema in the surrounding soft tissue.Conclusion:High resolution magnetic resonance imaging of the forefoot can demonstrate the anatomy and features of ligament and tendon injuries in forefoot accurately, which has clinical application value for the early diagnosis and accurate treatment of the injury.
5.Effects of fermented cordyceps powder on the expression of NLRP3 in myocardium of diabetic rats
Yuanjun LYU ; Zhuo WANG ; Zhuang CUI ; Jine LI ; Xiaofeng TAN ; Changping LI
Tianjin Medical Journal 2017;45(8):856-859,前插2
Objective To investigate the therapy effect of fermented cordyceps powder on diabetic cardiomyopathy, and the effect on expressions of NLRP3, Caspase-1 and interleukin (IL)-1βin myocardium of diabetic rats. Methods A total of 45 male SD rats were randomly divided into normal control group (group CON), diabetes group (group DM) and fermented cordyceps powder administration group (group CS, 1 g · kg-1 · d-1). Diabetes rat model was induced by tail vein injection with streptozotocin (45 mg/kg) in group DM and group CS. The rats in group CS were given fermented cordyceps powder solution by gavage at a single dose of 1 g·kg-1·d-1 for 18 weeks. Rats of group CON and group DM were given 0.9%NaCl in the same way. All rats were sacrificed after 18 weeks. The pathological changes in myocardium were observed by HE staining and electron microscope. The expression of NLRP3 in myocardium was detected by Western blotting. The expression levels of NLRP3, Caspase-1 and IL-1βin myocardium were detected by immunohistochemistry. Results (1) Myocardial necrosis and fibrosis and mitochondrial damages were observed under electron microscope in group DM. The above-mentioned pathological changes were improved in the group CS. (2) The result of Western blotting showed that the expression of NLRP3 in myocardium was significantly increased in group DM and group CS than that in group CON, and the expression of NLRP3 was significantly lower in group CS than that in group DM. (3) The result of immunohistochemistry showed that, compared with the group CON, the expressions of NLRP3, Caspase-1 and IL-1β in myocardium were significantly increased in group DM and group CS, and which were significantly lower in group CS than those in group DM. Conclusion Fermented cordyceps powder can improve diabetic cardiomyopathy through decreasing the expressions of NLRP3, Caspase-1 and IL-1βin myocardium.
6.Analysis of the Difference of β-Lactamase-Resistant Phenotypes by Pseudomonas Aeruginosa from Different Specimens
Jing WANG ; Wei CHEN ; Xiaoyan ZENG ; Yi ZHANG ; Jine LEI
Journal of Modern Laboratory Medicine 2017;32(3):63-66,70
Objective To compare clinical Pseudomonas aeruginosa different sources of β-lactamase-resistant phenotype differences,as to provide theoretical basis for guiding clinical rational use of antibiotics.Methods Isolation of 478 strains of Pseudomonas aeruginosa were collected from clinical specimens in the First Affiliated Hospital of Xi'an Jiaotong University from January to December 2015,by VITEK 2 Compact bacteria identification and drug sensitivity analysis of advanced expert system for β-lactamase-resistant phenotype,statistical analysis of drug resistance phenotype and antibiotic resistance.Results 478 strains of Pseudomonas aeruginosa were mainly composed of phenotype 5 and phenotype 3.Sputum,drainage fluid and bile duct bile specimens of Pseudomonas aeruginosa were based on phenotype 5,accounted for 31.08%,34.71% and 38.46%.Multiple comparison x2 were 3.893,4.071 and 5.595,There was no statistical difference between groups compare significance (P>0.05).Urine,secretions and whole blood samples of Pseudomonas aeruginosa were phenotype 3,accounted for 34.88 %,27.78 %,45.45 %;Multiple comparison x2 were 6.654,9.956 and 9.852.There was no statistical difference between groups compare significance (P>0.05).Sputum,drainage of liquid,bile duct bile and urine,secretion,whole blood specimens respectively source of Pseudomonas aeruginosa resistant phenotype distribution of two comparative difference was statistically significant (x2 =15.056~22.050,P<0.05).Comparing the resistance of different β-lactamase-resistant phenotypes in Pseudomonas aeruginosa isolates from different sources:the sputum specimen source in imipenem,meropenem,piperacillin and piperacillin/tazobactam had significant difference (x2 =22.225~39.025,P<0.05).There was statistical significance in department of hepatobiliary surgery only ceftazidime and meropenem differences (x2 =21.890~22.872,P<0.05).Conclusion The phenotypic analysis of β-lactamase-resistant phenotypes of Pseudomonas aeruginosa from different specimens was different,which provided a theoretical basis for guiding the clinical application of antibiotics and the control of nosocomial infection of Pseudomonas aeruginosa.
7.Engineering of Thiamin Pyrophosphate Fluorescent Biosensors Based on Ribozyme Switches in Mammalian Cells
Yuanyuan ZHANG ; Hui CHENG ; Yan SUN ; Jine WANG ; Zhengyan WU ; Renjun PEI
Chinese Journal of Analytical Chemistry 2017;45(2):157-162
Thiamin pyrophosphate (TPP) is a thiamine (vitamin B1) derivative and an essential cofactor in oxidative metabolism of the sugars,fatty acids and amino acids in living cells.By now,numerous TPP-dependent artificial riboswitch systems have been developed to regulate target gene expression but limited in bacteria,fungi or plant cells.Herein,the activating (switch-on) and inhibiting (switch-off) TPP-depended hammerhead ribozyme switches,which are from previous reported structures of prokaryotes screening,were investigated in mammalian cells.These ribozyme switches were inserted into the 3'UTR of the enhanced green fluorescence protein (EGFP) gene to construct the efficient ribozyme-based artificial switches through overlap extension PCR cloning.The HEK293 cells were transfected with the engineered ribozyme switches at increasing concentration of TPP.The EGFP gene-regulatory ability was analyzed with fluorescent microscope and flow cytometry.These TPP-inducible gene regulation devices showed the obvious ligand dose-dependency and excellent specificity.Two switch-on and one switch-off constructs demonstrated 3.1-fold or 1.9-fold increment and 2.3-fold reduction of EGFP level respectively with 150 μ mol/L TPP.The ligand-responsive ribozyme switches,by tuning the change of TPP concentration into the visual reporter genetic expression in cells,enable an efficient development of label-free,noninvasive and high-specific biosensors in living mammalian cells.
8.Clinical efficacy of raltitrexed combined with oxaliplatin and FOLFOX 4 protocol in treatment of patients with middle and advanced primary liver cancer
Yongqi SHEN ; Jun HUANG ; Chaoting CHEN ; Tao SI ; Zhixiang WANG ; Huadong XIE ; Xiangying KONG ; Jine LIU ; Chaowen HAN
Journal of Clinical Medicine in Practice 2017;21(7):39-42,46
Objective To explore the clinical efficacy and drug-toxic reactions of raltitrexed combined with oxaliplatin (RALOX protocol) and 5-fluorouracil + calciumfolinate + oxaliplatin (FOLFOX 4 protocol) in the treatment of patients with middle and advanced primary liver cancer (PLC).Methods A total of 72 patients with PLC were selected and randomly divided into RALOX group (n =34) and FOLFOX 4 group (n =38).The objective response rate (RR) was evaluated every 6 weeks after chemotherapy,while objective remission rate (OR),disease-control rate (DCR),median survival rate (mOS),median progression-free survival (mPFS),1-year survival rate (SR) as well as toxic and adverse reactions were observed.Results In RALOX group,31 patients were evaluable,with OR,DCR,mOS,mPFS,and 1-year SR being 19.4%,51.6%,7.2 months,3.4 months,and 22.6%,respectively.In FOLFOX 4 group,29 patients were evaluable,with OR,DCR,mOS,mPFS,and 1-year SR being 13.8%,48.3%,6.9 months,3.3 months and 20.7%,respectively.RALOX group was significantly lower than FOLFOX 4 group in the incidence rates of gastrointestinal reactions,liver toxicity,cardiac toxicity,peripheral nervous toxicity and hand-foot syndrome,but there were no significant differences in the incidence rates of renal toxicity and myelosuppression between two groups.Conclusion RALOX is safe and effective in the treatment of patients with middle and advanced PLC,and is superior to FOLFOX 4 protocol in clinical efficacy with mild adverse reactions.
9.Clinical efficacy of raltitrexed combined with oxaliplatin and FOLFOX 4 protocol in treatment of patients with middle and advanced primary liver cancer
Yongqi SHEN ; Jun HUANG ; Chaoting CHEN ; Tao SI ; Zhixiang WANG ; Huadong XIE ; Xiangying KONG ; Jine LIU ; Chaowen HAN
Journal of Clinical Medicine in Practice 2017;21(7):39-42,46
Objective To explore the clinical efficacy and drug-toxic reactions of raltitrexed combined with oxaliplatin (RALOX protocol) and 5-fluorouracil + calciumfolinate + oxaliplatin (FOLFOX 4 protocol) in the treatment of patients with middle and advanced primary liver cancer (PLC).Methods A total of 72 patients with PLC were selected and randomly divided into RALOX group (n =34) and FOLFOX 4 group (n =38).The objective response rate (RR) was evaluated every 6 weeks after chemotherapy,while objective remission rate (OR),disease-control rate (DCR),median survival rate (mOS),median progression-free survival (mPFS),1-year survival rate (SR) as well as toxic and adverse reactions were observed.Results In RALOX group,31 patients were evaluable,with OR,DCR,mOS,mPFS,and 1-year SR being 19.4%,51.6%,7.2 months,3.4 months,and 22.6%,respectively.In FOLFOX 4 group,29 patients were evaluable,with OR,DCR,mOS,mPFS,and 1-year SR being 13.8%,48.3%,6.9 months,3.3 months and 20.7%,respectively.RALOX group was significantly lower than FOLFOX 4 group in the incidence rates of gastrointestinal reactions,liver toxicity,cardiac toxicity,peripheral nervous toxicity and hand-foot syndrome,but there were no significant differences in the incidence rates of renal toxicity and myelosuppression between two groups.Conclusion RALOX is safe and effective in the treatment of patients with middle and advanced PLC,and is superior to FOLFOX 4 protocol in clinical efficacy with mild adverse reactions.
10.Differences of brain mechanism between selective attention and sustained attention:evidences from event-related potentials
Chinese Journal of Tissue Engineering Research 2016;20(33):4993-4998
BACKGROUND:At present, researches about attention mainly concentrate on the relationship between attention and other psychological phenomena, but studies concerning different types of attention itself (sustained attention versus selective attention) are few. OBJECTIVE:To investigate the brain mechanisms of sustained and selective attentions which have different roles in the cognitive process. METHODS:Event-related potentials (ERPs) were measured when 18 col ege students’ participants (half male and half female) were performing selective and sustained attention tasks. The program was written in E-Prime and displayed on a computer, and the experimental task was adapted from Karl et al. reported experimental paradigm, including selective attention and sustained attention tasks. The time of fixation point given was 500 ms, and the time of three randomly abreast pictures given was 300 ms. Al the participants were asked to make rapid and accurate response as much as possible. RESULTS AND CONCLUSION:The ERPs elicited by the two attention tasks were different in the aspects of time process, wave crest and latency. The brain regions activated by the sustained attention were wider than those by selective attention. The ERPs elicited by the sustained attention mainly presents as positive waves, whereas those elicited by the selective attention were reflected as both positive and negative waves. The EPRs elicited by the sustained attention were N100 and N250, but in the metaphase, selective attention elicited the P180. These may indicate that event choice appears in the middle stage of the task. Besides, the reversion of EPRs elicited by the two attention tasks in the anterior and posterior scalp is likely to be a generality of al attentions.

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