1.The mechanism of SAP overexpression in alleviating periodontitis in mice
HUANG Yinyin ; LIANG Dongliang ; ZOU Yaokun ; HAN Jingru ; GE Qing ; LIU Xueyan ; GUO Yadong ; HUANG Xinli ; YANG Lan
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(8):619-630
Objective:
To investigate the mechanism by which serum amyloid P component (SAP) alleviates periodontitis in mice, providing an experimental basis to establish SAP as a novel therapeutic agent for periodontitis.
Methods:
Ethical approval was obtained from the Institutional Animal Ethics Committee. Periodontitis models were established in wild-type (WT) mice and SAP-transgenic (SAP-Tg) mice, divided into four groups: WT control (WT group), WT periodontitis (WT+P group), SAP-Tg control (Tg group), and SAP-Tg periodontitis (Tg+P group). On day 7, the mice were euthanized, and periodontal tissues, teeth, and alveolar bone were collected. SAP protein expression was detected by enzyme-linked immunosorbent assay (ELISA). Micro-CT and HE staining were used to measure alveolar bone resorption (distance from the cementoenamel junction to the alveolar bone crest). Tartrate-resistant acid phosphatase (TRAP) staining was performed to assess osteoclast number, and immunohistochemistry (IHC) was employed to evaluate macrophage infiltration. The expression levels of inflammatory cytokines including interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were measured by qRT-PCR. Oral microorganism composition was analyzed using 16S ribosomal RNA (16S rRNA) gene sequencing. Additionally, macrophages from WT and SAP-Tg mice were isolated to establish an in vitro inflammation model, divided into WT+LPS and Tg+LPS groups. The expression of macrophage polarization-related genes including inducible nitric oxide synthase (iNOS), CD86, CD163, and CD206) were assessed by qRT-PCR. After the induction of osteoclast differentiation, TRAP staining was performed.
Results:
ELISA results demonstrated that periodontal tissues from Tg+P group mice exhibited higher levels of SAP expression compared to the WT+P group. Micro-CT and HE staining analyses revealed that the Tg+P group showed reduced alveolar bone resorption, indicated by a shorter distance between the cementoenamel junction and alveolar bone crest, compared to the WT+P group. Furthermore, TRAP staining results indicated a decrease in osteoclast numbers in the Tg+P group compared to the WT+P group. IHC and qRT-PCR results indicated reduced macrophage infiltration and decreased expression of IL-1β, IL-6, and TNF-α in the Tg+P group. Oral microorganism sequencing showed no significant difference in periodontitis-associated pathogenic bacteria between WT+P and Tg+P groups. In vitro experiments demonstrated that compared to the WT+LPS group, the Tg+LPS group exhibited downregulated M1 macrophage markers (iNOS and CD86) and upregulated M2 macrophage markers (CD163 and CD206). TRAP staining confirmed fewer osteoclasts in the Tg+LPS group.
Conclusion
SAP overexpression effectively alleviates periodontitis severity in mice by inhibiting M1 macrophage polarization, reducing pro-inflammatory cytokine expression, and suppressing osteoclast differentiation, thereby attenuating alveolar bone resorption.
2.Application progress of multi-omics technology in clinical nursing research
Jianmei YANG ; Guiru LIN ; Wanyi OU ; Dongliang CHEN ; Yuanfei WANG ; Aiping WU ; Zumin CHEN ; Ze ZENG ; Xiaomin LU ; Chenli LIN ; Yinji LIANG
Chinese Journal of Nursing 2024;59(16):2044-2048,后插1
The update of multi-omics technology is a key means to promote the rapid development of accurate health model in the whole life cycle.It can formulate dynamic and accurate nursing measures and provide massive data information from the perspective of nursing biology of health and disease.At present,clinical nursing research faces many challenges such as insufficient application and transformation ability of multi-omics technology.This paper introduces the multi-omics technology,reviews the application status of multi-omics technology in cancer nursing,maternal and child nursing,chronic metabolic disease nursing and symptom management,and puts forward the cross integration and prospect of multi-omics technology and nursing research,so as to strengthen the information mining ability of nurses at different levels of health and disease,and provide an important basis for accelerating the clinical transformation of precision nursing.
3.A comparative study of pathological results of the transperineal and transrectal cognitive targeted prostate biopsy based on bpMRI
Dongliang CAO ; Yifei CHENG ; Feng QI ; Minjie PAN ; Linghui LIANG ; Lei ZHANG ; Gong CHENG ; Lixin HUA
Chinese Journal of Urology 2022;43(3):187-192
Objective:To compare the differences of prostate cancer (PCa) and clinically significant prostate cancer (CsPCa) positive rate and postoperative complications between transperineal cognitive prostate biopsy (COG-TPBx) and transrectal cognitive prostate biopsy (COG-TRBx) based on biparametric magnetic resonance imaging (bpMRI).Methods:The data of 276 patients undergoing prostate biopsy from January 2019 to June 2021 in the First Affiliated Hospital of Nanjing Medical University were retrospectively reviewed. 157 patients underwent COG-TPBx(TPBx group) and 119 patients underwent COG-TRBx (TRBx group). The average age [(66.39 ± 8.31) vs. (66.30 ± 8.42)years], body mass index (BMI) [(23.85±2.49) vs. (23.68±2.61) kg/m 2], PSA values [9.43(1.47-19.80) vs. 8.94(0.66-19.99) ng/ml], prostate volume [37.92(13.99-167.40) vs. 40.78(11.67-188.21) cm 3], PSA density [0.21(0.04-1.17) vs. 0.20(0.04-1.04) ng/(ml·cm 3)], and suspicious digital rectal examination [17.20% (27/157) vs. 21.10% (25/119) ] were not significantly different between TPBx group and TRBx group. The positive rate of PCa, CsPCa, as well as post-biopsy complications of the two groups were compared. Results:There were no significant differences in the positive rate of PCa [49.68%(78/157) vs. 47.06%(56/119), P=0.666] and CsPCa [38.22%(60/157) vs. 34.45%(41/119), P=0.520] between the two groups. In stratification analysis, TPBx group has a significantly higher positive rate of both PCa [54.69%(35/64)] and CsPCa[43.75%(28/64)] in apex zone than TRBx group[39.62%(21/53) and 20.75%(11/53), all P<0.05). Moreover, the postoperative complications were not significantly different in TPBx group compared to that in TRBx group [10.19% (16/157) vs. 12.61%(15/119), P= 0.567]. Conclusions:Our investigations revealed that the overall positive rate of PCa, CsPCa, and the complications were not statistically different between COG-TPBx and COG-TRBx. COG-TPBx has a significantly higher positive rate of both PCa and CsPCa in apex zone.
4.The value of utilizing bpMRI in prostate biopsy in the detection of prostate cancer with PSA≤20 ng/ml
Minjie PAN ; Feng QI ; Yifei CHENG ; Dongliang CAO ; Linghui LIANG ; Lei ZHANG ; Gong CHENG ; Lixin HUA
Chinese Journal of Urology 2021;42(1):18-22
Objective:To detect the value of utilizing bpMRI in prostate biopsy in the detection of prostate cancer with PSA≤20ng/ml.Methods:The clinical data of 394 patients who underwent prostate biopsy in the First Affiliated Hospital of Nanjing Medical University from November 2017 to October 2019 were retrospectively analyzed. Of all the patients, 177 underwent modified systematic biopsy, named TRUS group, 217 patients accepted pre-biopsy bpMRI examination, undergoing modified systematic biopsy if Prostate Imaging Reporting and Data System (PI-RADS) score < 3 or MRI-TRUS cognitive fusion targeted prostate + systematic biopsy if PI-RADS score ≥ 3, named MRI group. The median age of TRUS group was 66 (61, 74) years old, prostate specific antigen (PSA) was 9.52 (7.26, 12.30) ng / ml, and prostate volume (PV) was 36.84 (28.95, 57.72)ml. The median age of MRI group was 66 (59, 72) years old, PSA was 8.84 (6.65, 12.16) ng/ml, and PV was 39.45 (29.25, 58.69)ml. There was no difference in above parameters between the two groups. The χ 2 test was used to compare the detection rate of prostate cancer and clinically significant prostate cancer (CsPCa) between the two groups. Results:There was no significant difference in the detection rates of prostate cancer between TRUS group and MRI group [51.41% (91/177) vs. 48.39% (105/ 217), P = 0.550], but the detection rates of CsPCa were significantly different [26.55% (47/177) vs. 36.41% (79/217), P = 0.037]. In patients with PSA ≤ 10 ng / ml, there was no significant difference in the detection rates of prostate cancer between the two groups [43.62% (41/94) vs. 43.08% (56/130), P = 0.936], but there was a significant difference in the detection rates of CsPCa [17.02% (16/94) vs. 28.46% (37/130), P = 0.047]. There was no significant difference in the detection rates of prostate cancer [60.24% (50/83) and 56.17% (48/87), P= 0.504] and the detection rates of CsPCa [37.35% (31/83) vs. 48.28% (42/87), P = 0.150] between the two groups. The total detection rates of the last two needles in TRUS group and MRI group were 23.16% (41/177) and 36.63% (86/217), respectively, with significant difference ( P=0.001); the detection rates of CsPCa in the last two needles were 11.86% (26/177) and 29.03% (63/ 217), respectively, with significant difference ( P < 0.001). In MRI group, the detection rates of prostate cancer in patients with PI-RADS score <3, 3, 4, 5 were 21.21% (7/33), 25.84% (23/89), 73.24% (52/71), 95.83% (23/24), respectively; the detection rates of CsPCa were 12.12% (4/33), 17.98% (16/89), 54.93% (39/71), 83.33% (23/24), respectively. Conclusions:In patients with PSA ≤ 20 ng / ml, prostate biopsy based on bpMRI may improve the detection of CsPCa, especially in patients with PSA ≤ 10 ng/ml.
5.A comparative study of prostate cancer detection rate between transperineal cognitive fusion targeted biopsy and software fusion targeted biopsy
Yifei CHENG ; Linghui LIANG ; Feng QI ; Lei ZHANG ; Dongliang CAO ; Shangqian WANG ; Gong CHENG ; Lixin HUA
Chinese Journal of Urology 2020;41(9):661-666
Objective:To investigate the difference of prostate cancer (PCa) detection rate between transperineal cognitive fusion targeted biopsy (COG-TB) and software fusion targeted (FUS-TB).Methods:We retrospectively analyzed 157 patients accepted transperineal targeted biopsies from December 2018 to December 2019, including 67 cases of COG-TB and 90 cases of FUS-TB. All patients were prostate biopsy na?ve, with PSA levels ≤ 20 ng/ml and prostate imaging reporting and data system version 2.1 (PI-RADS v2.1) scores ≥ 3. There was no significant difference between COG-TB and FUS-TB in the age [(70.78 ± 8.86) vs. (70.52 ± 8.79) years old], body mass index [(24.36 ± 2.69)vs. (24.14 ± 3.15) kg/m 2], prostate volume [36.69 (27.52, 47.40) vs. 38.81 (28.80, 53.46) cm 3], PSA level [8.27 (6.0, 11.65) vs. 8.88 (6.40, 13.54) ng/ml], PSAD [0.23 (0.15, 0.36) vs. 0.21 (0.14, 0.34) ng/ml 2], suspicious digital rectal examination findings [16 (23.9%) vs. 17 (18.9%)] and PI-RADS scores [24 (35.8%), 24 (35.8%), 19 (28.4%) and 21 (23.3%) vs. 21 (23.3%), 42 (46.7%), 27 (30.0%) for PI-RADS 3, 4, and 5, respectively]. There was no significant difference in baseline characteristics between the two groups (all P<0.05). The overall and stratified detection rates of PCa and clinically significant prostate cancer (CsPCa) were compared between the two groups. The upgrading rates of Gleason score after radical prostatectomy against biopsy Gleason score were compared between the two groups. Results:There was no significant difference between COG-TB and FUS-TB in the detection rate of PCa [76.1% (51/67) vs. 68.9% (62/90), P=0.32]. Also, no significant difference was found in the detection rate of PCa stratified by PSA [0-10ng/ml: 69.1% (29/42) vs. 57.1% (28/49); 10-20ng/ml: 88.0% (22/25) vs. 82.9% (34/41); all P>0.05] and PI-RADS score [3: 45.8% (11/24) vs. 23.8% (5/21); 4: 91.7% (22/24) vs. 81.0% (34/42); 5: 94.7% (18/19) vs. 85.2% (23/27); all P>0.05]. Similarly, there was no dramatically difference between COG-TB and FUS-TB in the detection rate of CsPCa [58.2% (39//67) vs. 50.0% (45/90), P>0.05]. No significant difference was found in the detection of CsPCa stratified by PSA [0-10ng/ml: 45.2% (19/42) vs.36.7% (18/49); 10-20 ng/ml: 80.0% (20/25) vs. 65.9% (27/41) ; all P>0.05] and PI-RADS score [3: 29.2% (7/24) vs. 9.5% (2/21), 4: 66.7% (16/24) vs. 57.1% (24/42), 5: 84.2%(16/19) vs. 70.4% (19/27) ; all P>0.05]. Additionally, the two technique was not different significantly in the upgrading rate [28.9% (13/45) vs. 26.2% (11/42), P=0.78]. Conclusions:There is no significant difference between FUS-TB and COG-TB in the detection rate of PCa and CsPCa, along with the upgrading rate after RP in patients with PSA ≤ 20 ng / ml and PI-RADS v2.1 score≥3.
6.Use of bpMRI-based cognitive fusion targeted biopsy and systematic biopsy in patients with PI-RADS≥3
Feng QI ; Yifei CHENG ; Linghui LIANG ; Lei ZHANG ; Dongliang CAO ; Gong CHENG ; Lixin HUA
Chinese Journal of Urology 2020;41(11):840-845
Objective:To investigate the use of bi-parametric magnetic resonance imaging (bpMRI)-based cognitive fusion targeted biopsy and systematic biopsy in patients with prostate imaging reporting and data system (PI-RADS)≥3.Methods:The clinical data of 220 patients with PI-RADS ≥3 who underwent bpMRI-transrectal ultrasound (TRUS) cognitive fusion targeted biopsy and systematic biopsy in the First Affiliated Hospital of Nanjing Medical University from May 2018 to November 2019 were retrospectively analyzed. The median age was 66 (60, 73) years old, median prostate specific antigen (PSA) was 8.73 (6.52, 11.93) ng/ml, medlian prostate volume was 39.25(29.26, 58.39) ml and the mean body mass index (BMI) was (24.02±2.60) kg/m 2. For each patient, bpMRI-TRUS cognitive fusion targeted biopsy and systematic biopsy were performed by two independent experienced urologists. The primary endpoint was the detection rate of CsPCa-A [clinically significant prostate cancer-A, defined as International Society of Urological Pathology (ISUP) grade group 2 or higher tumors]. The secondary endpoints were the detection rates of CsPCa-B (defined as ISUP grade group 3 or higher tumors) and CIPCa (clinically insignificant prostate cancer, defined as ISUP grade group 1 tumors). McNemar test and Chi-square test were used to compare the positive rates of CsPCa-A, CsPCa-B and CIPCa between targeted biopsy and systematic biopsy. Results:In this study, 112 patients (50.91%) were diagnosed with prostate cancer, and the detection was 42.73% (94/220) in targeted biopsy and 46.82% (103/220) in systematic biopsy.CsPCa-A was detected in 84 (38.18%) patients. Detection of CsPCa-A by targeted biopsy and systematic biopsy was not different significantly [30.00% (66/220) vs.34.09% (75/220), P=0.120]. CsPCa-A would have been missed in 8.18% (18/220) patients had not performed systematic biopsy, and in 4.09% (9/220) patients had not performed targeted biopsy. CsPCa-B was detected in 26.36% (58/220) patients. Detection of CsPCa-B by targeted biopsy and systematic biopsy was not different significantly [20.00% (44/220) vs. 23.18% (51/220), P=0.190]. CsPCa-B would have been missed in 6.36% (14/220) patients had not performed systematic biopsy, and in 3.18% (7/220) patients had not performed targeted biopsy. In addition, there was no difference in the positive rates of CIPCa between targeted biopsy combined with systematic biopsy, targeted biopsy only or systematic biopsy only [all three were 12.73% (28/220), P=1.000]. Nine post-biopsy adverse events were reported, including 5 cases of infection, 2 cases of vagal reflex and 2 cases of urinary retention. All of them were improved after symptomatic treatment. Conclusions:No significant difference was identified in the detection rate of CsPCa between targeted biopsy and systematic biopsy. However, combination of targeted biopsy and systematic biopsy could further improve the detection rate of CsPCa without increasing the detection of CIPCa. Therefore, a pre-biopsy bpMRI did have significant importance in the biopsy-na?ve patients, but did not seem to skip the need for systematic biopsy.
7.Effects of Hippo signaling pathway on lung injury repair by mesenchymal stem cells in acute respiratory distress syndrome.
Lang LI ; Liang DONG ; Fei GAO ; Jiaojie HUI ; Yizhe CHEN ; Jie YAN
Chinese Critical Care Medicine 2019;31(3):281-287
OBJECTIVE:
To investigate the effects of Hippo signaling pathway on lung injury repair of mesenchymal stem cells (MSC) in acute respiratory distress syndrome (ARDS) and its mechanism.
METHODS:
Mouse bone marrow-derived MSC (mMSCs) cell lines with low expression of large tumor suppressor 2 (LATS2) were constructed by lentiviral vector transfection. Male C57BL/6 mice aging 6-8 weeks old were divided into four groups according to random number table (n = 36). The ARDS animal model (ARDS group) was reproduced by intratracheally injection of 2 g/L lipopolysaccharide (LPS) 50 μL, the normal saline (NS) control group was injected with an equal volume of NS. After 4 hours of model reproduction, 5×104 mMSCs transfected with blank lentivirus vector (MSC-shcontrol group) or shLATS2 lentivirus vector (MSC-shLATS2 group) were transplanted intratracheally, while NS control group and ARDS group were injected with equal volume of phosphate buffered saline (PBS). Mice were sacrificed at 3, 7, and 14 days after modeling, and lung tissue and bronchoalveolar lavage fluid (BALF) were harvested. Near-infrared fluorescence imaging, immunofluorescence staining and Western Blot were used to track mMSCs in lung tissue. Retension and proportion of mMSC differentiation into type II alveolar epithelial cells (AEC II) were evaluated. Lung tissue wet weight/body weight ratio (LWW/BW) and total protein (TP) and albumin (ALB) in BALF were determined to reflect pulmonary edema. The expression of Occludin protein in lung epithelium was tested by Western Blot to reflect permeability of epithelium. The levels of interleukins (IL-1β, IL-6, IL-10) in BALF were assessed by enzyme-linked immunosorbent assay (ELISA) to reflect lung inflammation. Hematoxylin-eosin (HE) staining and modified Masson staining were carried out, and the scores were assessed to reflect lung injury and evaluate pulmonary fibrosis.
RESULTS:
The signal intensity of isolated lung fluorescence images at 3 days in the MSC-shLATS2 group was significantly higher than that in the MSC-shcontrol group (fluorescence intensity: 0.039±0.005 vs. 0.017±0.002, P < 0.05), the number of green fluorescent protein (GFP)-positive cells in lung tissue was also significantly higher than that in the MSC-shcontrol group (cells/HP: 29.65±6.98 vs. 17.50±4.58, P < 0.05), but they all decreased with time; and the proportion of mMSCs differentiated into AEC II was significantly increased [(64.12±15.29)% vs. (19.64±3.71)%, P < 0.05]. Compared with the NS control group, the levels of surface active protein C (SPC) and Occludin protein in the ARDS group were significantly decreased, LWW/BW ratio and TP, ALB and inflammatory factors levels in BALF were significantly increased; extensive alveolar and interstitial edema, hemorrhage and diffuse inflammatory cell infiltration were found in lung tissue, and the lung injury score was significantly increased; collagen fibers were deposited in alveolar septum and alveolar cavity, and pulmonary fibrosis score was also increased significantly. Compared with the ARDS group, the expression levels of SPC and Occludin at 14 days in the MSC-shcontrol group and the MSC-shLATS2 group were significantly increased (SPC/β-actin: 0.51±0.12, 0.68±0.10 vs. 0.27±0.08, Occludin/β-actin: 0.49±0.19, 0.79±0.11 vs. 0.25±0.08, all P < 0.05), TP, ALB, IL-1β, IL-6 levels in BALF at 3 days were significantly decreased [TP (g/L): 8.08±1.72, 5.12±0.87 vs. 12.55±2.09; ALB (g/L): 0.71±0.21, 0.44±0.18 vs. 1.18±0.29, IL-1β (ng/L): 99.26±14.32, 60.11±8.58 vs. 161.86±25.17, IL-6 (ng/L): 145.54±13.29, 101.74±11.55 vs. 258.79±27.88, all P < 0.05], and IL-10 was significantly increased (ng/L: 190.83±22.61, 316.65±37.88, both P < 0.05). Furthermore, all the above parameters in the MSC-shLATS2 group were significantly improved as compared with those in the MSC-shcontrol group (all P < 0.05). LWW/BW ratio in the MSC-shLATS2 group was significantly lower than that in the ARDS group and the MSC-shcontrol group (mg/g: 9.85±1.51 vs. 16.78±1.92, 14.88±1.74, both P < 0.05).
CONCLUSIONS
Inhibiting Hippo signaling pathway by low expression of LATS2 could promote the retention of mMSCs in lung tissue and differentiation into AEC II cells of ARDS mice, improve pulmonary edema and alveolar epithelial permeability, regulate pulmonary inflammatory response, and alleviate pathological damage and fibrosis of lung tissue.
Animals
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Hippo Signaling Pathway
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Lung Injury/prevention & control*
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Male
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Mesenchymal Stem Cells/metabolism*
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Mice
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Mice, Inbred C57BL
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Protein Serine-Threonine Kinases/metabolism*
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Respiratory Distress Syndrome/metabolism*
;
Signal Transduction
8.Large tumor suppressor gene 2-mediated Hippo signaling pathway regulates the biological behavior of mesenchymal stem cells in vitro.
Chinese Critical Care Medicine 2019;31(9):1143-1148
OBJECTIVE:
To investigate the regulatory effect of Hippo signaling pathway mediated by large tumor suppressor gene 2 (LATS2) on biological behavior of mice bone marrow mesenchymal stem cells (BMSCs) in vitro.
METHODS:
BMSCs of C57BL/6 mice were cultured in vitro and passed to the 6th to 7th generations for experiment. BMSCs with activated and inactivated LATS2 were constructed with lentiviral vectors transfections. The BMSCs were allocated to blank control group (MSC group), empty vector control group (MSC-eGFP group), LATS2-overexpressing group (MSC-LATS2 group), empty vector without LATS2 shRNA control group (MSC-shcontrol group) and LATS2-underexpressing group (MSC-shLATS2 group). The transduction efficiencies mediated by the lentiviral vectors were evaluated by flow cytometry. The mRNA and protein expressions of LATS2, phosphorylation of Yes associated protein (p-YAP) and 14-3-3 were quantified by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot respectively. Osteogenic and adipogenic differentiation of BMSCs were evaluated through alizarin red and oil red O staining. Proliferation of BMSCs was evaluated using the CCK-8 assay. The effect of Hippo pathway on horizontal and vertical migration ability of BMSCs was measured by the scratch test and Transwell chamber test.
RESULTS:
The transduction efficiencies mediated by the lentiviral vectors were 93.1%-97.1%. Compared with MSC-eGFP group, the expressions of LATS2, p-YAP and 14-3-3 in MSC-LATS2 group were significantly elevated [LATS2 mRNA (2-ΔΔCT): 2.55±0.13 vs. 1.08±0.05, LATS2/GAPDH: 2.63±0.11 vs. 1.06±0.08, p-YAP/total YAP: 1.67±0.11 vs. 1.00±0.04, 14-3-3/β-actin: 2.22±0.20 vs. 0.98±0.06, all P < 0.05], however, compared with MSC-shcontrol group, the expressions in MSC-shLATS2 group were significantly reduced [LATS2 mRNA (2-ΔΔCT): 0.10±0.01 vs. 1.01±0.05, LATS2/GAPDH: 0.09±0.01 vs. 1.05±0.06, p-YAP/total YAP: 0.10±0.02 vs. 1.10±0.09, 14-3-3/β-actin: 0.05±0.01 vs. 0.90±0.08, all P < 0.05]. It suggested that high and low expression of LATS2 could activate or inhibit Hippo pathway. The osteogenic and adipogenic differentiation and proliferation rate of BMSCs in MSC-LATS2 group were significantly lower than those in MSC-eGFP group, however, those in MSC-shLATS2 group were significantly higher than MSC-shontrol group (all P < 0.05). It suggested that high and low expression of LATS2 could inhibit or promote osteogenesis, adipogenesis and cell proliferation of BMSCs. Scratch test and Transwell chamber test showed that the degree of scratch healing in MSC-LATS2 group was significantly lower than that in MSC-eGFP group [(22.11±3.02)% vs. (45.99±6.58)%], while the number of cells migrating to the subventricular layer of Transwell was significantly reduced (cells/MP: 20.82±3.05 vs. 111.33±13.28, both P < 0.05); the degree of scratch healing in MSC-shLATS2 group was significantly higher than that in MSC-shcontrol group [(70.32±7.17)% vs. (39.28±2.98)%], the number of cells migrating to the subventricular layer of Transwell was increased significantly (cells/MP: 206.19±30.58 vs. 120.10±25.10, both P < 0.05). It suggested that high and low expression of LATS2 could inhibit or promote the horizontal and vertical migration of BMSCs.
CONCLUSIONS
LATS2-mediated alteration of Hippo pathway could modulate differentiation, proliferation, and migration of mesenchymal stem cells in vitro.
Animals
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Genes, Tumor Suppressor
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Mesenchymal Stem Cells
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Mice
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Mice, Inbred C57BL
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Protein Serine-Threonine Kinases
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Signal Transduction
9.miRNA-24-3P affect RAW264.7 cell function by modulating SP1
Dongliang CHENG ; Yuan LIANG ; Yanchen CHENG ; Di QING
Chongqing Medicine 2018;47(17):2257-2260,2265
Objective To discuss the mechanism of miR-24-3P in the process of acute respiratory distress syndrome (ARDS).Methods miR-24-3P mimics,miR-24-3P inhibitor and their negative controls were transfected in RAW264.7 cells respectively:The relative expression of miR-24-3P in each cell was detected.The relative expression of TNF-α,IL-6,SP1 and NF-κB were detected in each cell,and the regulation of miR-24-3P on the target gene SP1 was detected by dual luciferase reporter gene.Results The expression level of TNF-α mRNA and IL-6 mRNA in each group was statistically significant (P<0.05).The relative expression level of SP1 mRNA and NF-κB mRNA in each group was not statistically significant (P>0.05).The expression of SP1 in each group was statistically significant (P<0.05).The gene analysis of dual luciferase reporter showed that the fluorescence activity was significantly inhibited after cotransfection of miR-24-3P and SP1 in HEK293 cells(P<0.05).Conclusion miR-24-3P can play an important role in the process of ARDS by influencing the translation of SP1 gene and affecting the release of inflammatory mediators.
10.Relationship between MIF expression and obese-induced abolition of sevoflurane preconditioning-in-duced cardioprotection in mice
Xiyan ZHANG ; Fanqing MENG ; Li ZHANG ; Dongliang LI ; Jiali WANG ; Qiya HU ; Bingfeng SONG ; Liang LI
Chinese Journal of Anesthesiology 2018;38(8):911-915
Objective To evaluate the relationship between macrophage migration inhibitory factor ( MIF) expression and obese-induced abolition of sevoflurane preconditioning-induced cardioprotection in mice. Methods Forty-eight male C57BL∕6J mice, aged 4 weeks, were divided into 2 groups ( n=24 each) using a random number table method: normal diet group ( Lean group ) and high-fat diet group ( Obese group) . Lean group were fed a normal diet ( 10% kcal) for 12 weeks, while Obese group were fed a high-fat diet ( 60% kcal) for 12 weeks. The weight of mice was measured. Blood samples were collected from the tail vein for determination of blood glucose concentrations, and plasma concentrations of total cho-lesterol, triglyceride, insulin and leptin. After measurement of the parameters mentioned above, Lean group and Obese group were divided into 3 subgroups ( n=8 each) using a random number table method:sham operation groups (L-Sham group, O-Sham group), myocardial ischemia-reperfusion groups (L-IR group, O-IR group) and sevoflurane preconditioning groups (L-IR+Sev group, O-IR+Sev group). The mice were anesthetized and their hearts were immediately removed and retrogradely perfused in a Langendorff apparatus with an oxygenated K-H solution at 37 ℃. Hearts were continuously perfused with K-H solution for 115 min in L-Sham and O-Sham groups. Hearts were subjected to global ischemia for 25 min, followed by 60-min reperfusion after being retrogradely perfused with K-H solution in L-IR and O-IR groups. In L-IR+Sev and O-IR+Sev groups, hearts were subjected to 3 cycles of 5-min perfusion with sevoflurane-contai-ning K-H solution ( final concentration 0. 6 mmol∕L) and 5-min washout, and then hearts were subjected to global ischemia for 25 min, followed by 60-min reperfusion. Left ventricular developed pressure ( LVDP ) , left ventricular end-diastolic pressure ( LVEDP ) , and the maximum rate of increase or decrease in left ventricular pressure ( ±dp∕dtmax) were recorded at the end of reperfusion. Hearts were obtained at the end of reperfusion for determination of myocardial infarct size and expression of MIF ( by Western blot) . Results Compared with Lean group, the weight, blood glucose, levels of plasma total cholesterol, tri-glyceride, insulin and leptin were significantly increased in Obese group (P<0. 05). Compared with L-Sham group, the LVDP and +dp∕dtmax were significantly decreased, LVEDP and -dp∕dtmax were in-creased, myocardial infarct size was increased, and the expression of myocardial MIF was up-regulated in L-IR and L-IR+Sev groups, and the expression of myocardial MIF was up-regulated in O-Sham group ( P<0. 05) . Compared with L-IR group, LVDP and +dp∕dtmax were significantly increased, LVEDP and-dp∕dtmax were decreased, myocardial infarct size was decreased, and the expression of myocardial MIF was up-regulated in group L-IR+Sev, and the expression of myocardial MIF was significantly up-regulated in group O-IR (P<0. 05). Compared with O-Sham group, LVDP and +dp∕dtmax were significantly de-creased, LVEDP and-dp∕dtmax were increased, and myocardial infarct size was increased, and no signif-icant change was found in the expression of MIF in O-IR and O-IR+Sev groups ( P>0. 05) . Conclusion The mechanism by which obese abolishes sevoflurane preconditioning-induced cardioprotection may be relat-ed to inducing MIF over-expression in mice.


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