1.Isolation and identification of placental exosomes from maternal serum
Yujing LI ; Zhenyu DIAO ; Pingping XUE ; Li SHEN ; Ping GONG ; Guijun YAN ; Yali HU
Journal of Medical Postgraduates 2015;(6):632-636
Objective During pregnancy , exosomes can be released from the placenta into maternal circulation and play im-portant roles in normal pregnancy or placenta-related diseases .We aimed to establish a simple and efficient method for isolating and i-dentifying placental exosomes from maternal serum and lay a foundation for the studies of pregnancy -related diseases . Methods Using sucrose gradient centrifugation with 8% PEG6000 precipitation twice , we isolated and purified placenta-derived exosomes from normal maternal serum and detected their molecular markers CD 63 , CD81 and PLAP by Western blot , followed by silver staining anal-ysis of the protein profile of the exosome pellet .We identified the morphology of the placenta-derived exosomes by transmission electron microscopy ( TEM) and measured the size and distribution of the particles by dynamic light scattering ( DLS) . Results Silver stai-ning of the protein profiles of the exosomes after sucrose gradient centrifugation clearly revealed the bands of the protein molecules . Western blot showed the expressions of CD 63, CD81, and PLAP in the 21-34%density layer, which demonstrated the presence of serum placental exosomes mainly in the 1.09-1.16 g/mL density layer.TEM exhibited that the placenta-derived exosomes were round or oval cup-shaped, specifically expressing PLAP, and the particles were uniform in size, with a mean diameter of (41.79 ±11.94) nm. Conclusion A simple, fast, and efficient method was successfully established for isolating placenta-derived exosomes from ma-ternal serum, which provides a basis for studying the roles of placental exosomes in normal pregnancy and placenta -related diseases.
2.Effectiveness of exercise intervention on sleep quality and negative emotion among female college students with anxiety
GONG Yujing, ZHANG Yifan, YANG Xinyi, GUAN Yuan, TANG Donghui
Chinese Journal of School Health 2019;40(4):542-545
Objective:
To investigate the effect of exercise intervention on sleep quality of female college students with anxiety, and to further explore the mediating effect of negative emotions in this association.
Methods:
Seventy female college students were randomly divided into experimental group (34 patients) and control group(36 patients). The experimental group received eight-week aerobic exercise and yoga training. The control group received no training. Sleep Quality Scale, Negative Emotion Scale and Anxiety Self-rating Scale were used to assess the level of sleep and negative emotion before and after exercise intervention.
Results:
After exercise intervention, sleep quality (4.31±1.26), anxiety (36.41±7.32) and negative emotion (2.37±0.50) in the experimental group significantly improved(P<0.01), no similar changes were found in the control group(P<0.05). By examining the mediating effects, negative emotions played a partial mediating role in the association between exercise and sleep quality(t=6.77, P<0.01).
Conclusion
Exercise intervention significantly improved the quality of sleep and negative emotion among female college students with anxiety. Negative emotions play a partial role in the positive effect of exercise on sleep quality.
3.Heme oxygenase-1 reduces inflammatory response by inhibiting thioredoxin interacting protein/NOD-like receptor protein 3 inflammasome activation in RAW264.7 cells
Xinyue GAO ; Rong BAO ; Yujing FENG ; Zhaozhao HUANG ; Zhan GONG ; Yujie ZHOU
Chinese Critical Care Medicine 2022;34(7):689-692
Objective:To investigate the inhibitory effect and mechanism of heme oxygenase-1 (HO-1) on the inflammatory response of macrophages.Methods:Mouse macrophage strain RAW264.7 was cultured in vitro, and the cells in the logarithmic growth phase were used for the experiment. The RAW264.7 cells were divided into four groups. In blank control group, the cells were continuously incubated and received no treatment (cultured at 37 ℃, 95% air, 5% CO 2). In lipopolysaccharide (LPS) model group, 1 mg/L LPS was added to the medium to prepare LPS challenge model. In HO-1 inducer group, the cells were incubated with 30 μmol/L HO-1 inducer hemin for 1 hour, and then 1 mg/L LPS was added for incubation. In HO-1 inhibition group, the cells were incubated with 5 μmol/L HO-1 specific antagonist Zinc protoporphyrin Ⅸ (ZnPPⅨ) for 0.5 hour, and then 1 mg/L LPS was added for incubation. After 48 hours of incubation with LPS, the supernatant of each group was taken, and the protein expressions of HO-1, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), thioredoxin interacting protein (TXNIP), NOD-like receptor protein 3 (NLRP3) and mitochondrial autophagy marker microtubule-associated protein 1 light chain 3B (LC-3B) were detected by Western blotting. The expression of reactive oxygen species (ROS) was detected by immunofluorescence staining. Results:Compared with the blank control group, the cells in the LPS model group had a certain stress response, and autophagy occurred in mitochondria, but the expression of some inflammatory factors was restricted, which was related to the impairment of cell function. The protein expressions of HO-1, IL-1β, LC-3B, ROS were significantly increased, the protein expressions of TNF-α, TXNIP, and NLRP3 were decreased significantly, indicating that the cells were seriously injured after LPS challenge, and the model was successfully established. Compared with the LPS model group, HO-1 protein expression in the HO-1 inducer group was significantly increased (HO-1/GAPDH: 0.31±0.03 vs. 0.22±0.03, P < 0.05), the protein expressions of TNF-α, IL-1β, TXNIP, NLRP3, LC-3B and ROS were significantly inhibited [TNF-α protein (TNF-α/GAPDH): 0.08±0.01 vs. 0.45±0.05, IL-1β protein (IL-1β/GAPDH): 0.50±0.01 vs. 0.82±0.03, TXNIP protein (TXNIP/GAPDH): 0.21±0.02 vs. 0.28±0.02, NLRP3 protein (NLRP3/GAPDH): 0.11±0.01 vs. 0.17±0.02, LC-3B protein (LC-3B/GAPDH): 0.67±0.04 vs. 0.92±0.12, ROS (fluorescence intensity): 80.9±12.5 vs. 94.1±19.5, all P < 0.05], indicating that HO-1 could inhibit inflammatory response and oxidative stress, and reduce mitochondrial autophagy. Antagonizing HO-1 could increase inflammatory response, oxidative stress and mitochondrial autophagy, the inhibitory degree of TNF-α and IL-1β expression was significantly reduced as compared with the HO-1 inducer group [TNF-α protein (TNF-α/GAPDH): 0.26±0.02 vs. 0.08±0.01, IL-1β protein (IL-1β/GAPDH): 0.76±0.01 vs. 0.50±0.01, both P < 0.05], the protein expressions of TXNIP, NLRP3, LC-3B and ROS were significantly increased as compared with the LPS model group [TXNIP protein (TXNIP/GAPDH): 0.43±0.02 vs. 0.28±0.02, NLRP3 protein (NLRP3/GAPDH): 0.24±0.02 vs. 0.17±0.02, LC-3B protein (LC-3B/GAPDH): 1.12±0.07 vs. 0.92±0.12, ROS (fluorescence intensity): 112.0±17.0 vs. 94.1±19.5, all P < 0.05]. Conclusion:HO-1 can reduce the inflammatory response by inhibiting the activation of TXNIP/NLRP3 inflammasome and reducing the release of inflammatory mediators.
4.Clinicopathological analysis of 61 patients with rectal gastrointestinal stromal tumors.
Xiaojun WU ; Wu JIANG ; Rongxin ZHANG ; Peirong DING ; Gong CHEN ; Zhenhai LU ; Liren LI ; Yujing FANG ; Fulong WANG ; Lingheng KONG ; Junzhong LIN ; Zhizhong PAN ; Desen WAN
Chinese Journal of Gastrointestinal Surgery 2014;17(4):335-339
OBJECTIVETo explore the clinicopathological characteristics, efficacy, and prognostic factors for patients with rectal gastrointestinal stromal tumor(GIST).
METHODSClinicopathological and follow-up data of 61 patients with rectal GIST in our department from January 1990 to October 2012 were analyzed retrospectively and pathology specimens were reviewed. Kaplan-Meier method was used to calculate the survival. Univariate analysis and multivariate analysis were performed to investigate the influencing factors of prognosis with Log-rank test and Cox regression model.
RESULTSThere were 42 male and 19 female patients with a median age of 59 years old. Eighteen cases(29.5%) were confirmed preoperatively as GIST by biopsy and 46 cases were diagnosed as GIST by first pathological examination. Fifteen cases(24.6%) were revised as GIST after re-examination of specimes among whom 14 cases had been diagnosed as leiomyoma or sarcoma, and 1 as neurolemmoma. Tumor location was above peritoneal reflection in 12 cases(19.7%) and below peritoneal reflection in 49(80.3%). Fifty-two patients underwent surgery, including 21 extended resections(lymph nodes clearance and combined organs resection simultaneously) and 31 local resections(tumor rejection or partial resection of rectal wall). Eleven patients received preoperative imatinib(400 mg/d). Forty-one cases received imatinib therapy after operation or biopsy diagnosis, including 25 cases who received palliative treatment for postoperative recurrence. Median follow-up time was 55(6 to 391) months and follow-up longer than 2 years was carried out in 46 patients. Overall survival rates of 1-, 2-, 3- , 5-year were 98%, 95.6%, 86.0% and 73.7% respectively. There were no significant differences between local resection group(96.4%, 92%, 83.3% and 77.3%) and extended resection group (100%, 94.7%, 89.50% and 82.6%)(χ(2)=0.004, P=0.947). Univariate analysis showed that survival was only associated with recurrence and metastasis (χ(2)=4.292, P=0.038). Multivariate Cox analysis showed postoperative survival was not associated with any factors(all P>0.05). The 3-year survival rate of patients with postoperative recurrence or metastasis receiving imatinib therapy was better as compared to those who did not received imatinib(82.7% vs. 71.4%).
CONCLUSIONSRectal GIST are more common in the lower rectum. Surgery is the main treatment for rectal GIST. Local complete resection is the mainstay treatment. Extensive resection and lymph node clearance may not improve survival. Imatinib can improve the prognosis of patients with recurrence or metastasis.
Benzamides ; Female ; Gastrointestinal Stromal Tumors ; therapy ; Humans ; Imatinib Mesylate ; Male ; Neoplasm Recurrence, Local ; Piperazines ; Prognosis ; Pyrimidines ; Rectal Neoplasms ; pathology ; therapy ; Retrospective Studies ; Survival Rate
5.Effects of early postoperative enteral nutrition in combination of micro-ecological feed preparations on intestinal flora and immune function in elderly colorectal cancer patients after surgery
Jiefeng LIU ; Miao HE ; Xinyu ZENG ; Yujing GONG
Chinese Journal of Geriatrics 2020;39(4):435-438
Objective:To investigate effects of early postoperative enteral nutrition in combination of micro-ecological feed preparations on intestinal flora and immune function in elderly colorectal cancer patients after surgery.Methods:Eighty-eight elderly colorectal cancer patients treated at our hospital from November 2017 to August 2019 were enrolled in the study.They were randomly divided into two groups(n=44, each): the observation group receiving micro-ecological feed preparations and enteral nutrition, and the control group receiving conventional parenteral nutrition.The intestinal flora index, cellular immune function, humoral immune function and complications were compared between the two groups.Results:There was no significant difference in the indices of intestinal flora between the two groups before operation(all P>0.05). In the control group, Bifidobacteria and Lactobacillus decreased, E. coli increased, and B/E value decreased after surgery compared with before surgery( P<0.05). In the observation group, Bifidobacteria and Lactobacillus decreased, E. coli increased and B/E value decreased after surgery compared with before surgery( P<0.05). Compared with the control group, the observation group showed that Bifidobacteria and Lactobacillus increased, E. coli decreased and B/E value increased after surgery( P<0.05). Serum inflammatory factors had no statistically significance difference between the two groups before operation( P>0.05). Levels of interleukin-6, C-reactive protein and tumor necrosis factor-α were increased in the two groups after surgery compared with before surgery( P<0.05), and the indexes were lower in the observation group than in the control group( t=4.841, 7.247 and 4.231, all P=0.000). Levels of IgG, IgM, IgA, CD3 + , CD4 + , CD8 + and the ratio of CD4 + and CD8 + were higher in the observation group than in the control group[(12.53±1.84) g/L vs.(11.47±1.92)g/L, (1.45±0.22) g/L vs.(1.33±0.17) g/L, (2.28±0.24) g/L vs.(2.13±0.12) g/L, (52.84±4.12)% vs.(50.43±3.91)% , (34.53±3.51)% vs.(30.75±3.62)%, (19.09±2.41)% vs.(18.24±0.53)% and (1.68±0.22)% vs.(1.43±0.15)%, t=2.644, 2.863, 3.120, 2.814, 4.973, 2.285 and 6.228, P=0.010, 0.005, 0.003, 0.006, 0.000, 0.025 and 0.000]. Conclusions:Early micro-ecological feed preparation in combination of enteral nutrition can improve the surgical effect and quality of life and promote disease recovery with a safety in patients with colorectal cancer.
6.Clinicopathological analysis of 61 patients with rectal gastrointestinal stromal tumors
Xiaojun WU ; Wu JIANG ; Rongxin ZHANG ; Peirong DING ; Gong CHEN ; Zhenhai LU ; Liren LI ; Yujing FANG ; Fulong WANG ; Lingheng KONG ; Junzhong LIN ; Zhizhong PAN ; Desen WAN
Chinese Journal of Gastrointestinal Surgery 2014;(4):335-339
Objective To explore the clinicopathological characteristics, efficacy, and prognostic factors for patients with rectal gastrointestinal stromal tumor (GIST). Methods Clinicopathological and follow-up data of 61 patients with rectal GIST in our department from January 1990 to Oc tober 2012 were analyzed retrospectively and pathology specimens were reviewed. Kaplan-Meier method was used to calculate the survival. Univariate analysis and multivariate analysis were performed to investigate the influencing factors of prognosis with Log-rank test and Cox regression model. Results There were 42 male and 19 female patients with a median age of 59 years old. Eighteen cases (29.5%) were confirmed preoperatively as GIST by biopsy and 46 cases were diagnosed as GIST by first pathological examination. Fifteen cases (24.6%) were revised as GIST after re-examination of specimes among whom 14 cases had been diagnosed as leiomyoma or sarcoma, and 1 as neurolemmoma. Tumor location was above peritoneal reflection in 12 cases (19.7%) and below peritoneal reflection in 49 (80.3%). Fifty-two patients underwent surgery, including 21 extended resections(lymph nodes clearance and combined organs resection simultaneously) and 31 local resections (tumor rejection or partial resection of rectal wall). Eleven patients received preoperative imatinib(400 mg/d). Forty-one cases received imatinib therapy after operation or biopsy diagnosis, including 25 cases who received palliative treatment for postoperative recurrence. Median follow-up time was 55 (6 to 391) months and follow-up longer than 2 years was carried out in 46 patients. Overall survival rates of 1-, 2-, 3-, 5-year were 98%, 95.6%, 86.0%and 73.7%respectively. There were no significant differences between local resection group (96.4%, 92%, 83.3%and 77.3%) and extended resection group (100%, 94.7%, 89.50%and 82.6%) (χ2=0.004, P=0.947). Univariate analysis showed that survival was only associated with recurrence and metastasis (χ2=4.292, P=0.038). Multivariate Cox analysis showed postoperative survival was not associated with any factors (all P>0.05). The 3-year survival rate of patients with postoperative recurrence or metastasis receiving imatinib therapy was better as compared to those who did not received imatinib (82.7%vs. 71.4%). Conclusions Rectal GIST are more common in the lower rectum. Surgery is the main treatment for rectal GIST. Local complete resection is the mainstay treatment. Extensive resection and lymph node clearance may not improve survival. Imatinib can improve the prognosis of patients with recurrence or metastasis.
7.Clinicopathological analysis of 61 patients with rectal gastrointestinal stromal tumors
Xiaojun WU ; Wu JIANG ; Rongxin ZHANG ; Peirong DING ; Gong CHEN ; Zhenhai LU ; Liren LI ; Yujing FANG ; Fulong WANG ; Lingheng KONG ; Junzhong LIN ; Zhizhong PAN ; Desen WAN
Chinese Journal of Gastrointestinal Surgery 2014;(4):335-339
Objective To explore the clinicopathological characteristics, efficacy, and prognostic factors for patients with rectal gastrointestinal stromal tumor (GIST). Methods Clinicopathological and follow-up data of 61 patients with rectal GIST in our department from January 1990 to Oc tober 2012 were analyzed retrospectively and pathology specimens were reviewed. Kaplan-Meier method was used to calculate the survival. Univariate analysis and multivariate analysis were performed to investigate the influencing factors of prognosis with Log-rank test and Cox regression model. Results There were 42 male and 19 female patients with a median age of 59 years old. Eighteen cases (29.5%) were confirmed preoperatively as GIST by biopsy and 46 cases were diagnosed as GIST by first pathological examination. Fifteen cases (24.6%) were revised as GIST after re-examination of specimes among whom 14 cases had been diagnosed as leiomyoma or sarcoma, and 1 as neurolemmoma. Tumor location was above peritoneal reflection in 12 cases (19.7%) and below peritoneal reflection in 49 (80.3%). Fifty-two patients underwent surgery, including 21 extended resections(lymph nodes clearance and combined organs resection simultaneously) and 31 local resections (tumor rejection or partial resection of rectal wall). Eleven patients received preoperative imatinib(400 mg/d). Forty-one cases received imatinib therapy after operation or biopsy diagnosis, including 25 cases who received palliative treatment for postoperative recurrence. Median follow-up time was 55 (6 to 391) months and follow-up longer than 2 years was carried out in 46 patients. Overall survival rates of 1-, 2-, 3-, 5-year were 98%, 95.6%, 86.0%and 73.7%respectively. There were no significant differences between local resection group (96.4%, 92%, 83.3%and 77.3%) and extended resection group (100%, 94.7%, 89.50%and 82.6%) (χ2=0.004, P=0.947). Univariate analysis showed that survival was only associated with recurrence and metastasis (χ2=4.292, P=0.038). Multivariate Cox analysis showed postoperative survival was not associated with any factors (all P>0.05). The 3-year survival rate of patients with postoperative recurrence or metastasis receiving imatinib therapy was better as compared to those who did not received imatinib (82.7%vs. 71.4%). Conclusions Rectal GIST are more common in the lower rectum. Surgery is the main treatment for rectal GIST. Local complete resection is the mainstay treatment. Extensive resection and lymph node clearance may not improve survival. Imatinib can improve the prognosis of patients with recurrence or metastasis.