1.Comparison of Wrist Arthroscopy Assisted Closed Reduction With External Fixation and Open Reduction With Steel Plate Internal Fixation for Intra-articular Comminuted Fractures of the Distal Radius
Zizhen LIU ; Guangxin CHEN ; Bin TANG ; Zhaoxin LUAN ; Zhengxun LI
Chinese Journal of Minimally Invasive Surgery 2025;25(9):550-556
Objective To explore the clinical efficacy of wrist arthroscopy assisted closed reduction with external fixation in the treatment of intra-articular comminuted fractures of the distal radius(type C3).Methods A retrospective analysis was conducted on 98 patients with type C3 intra-articular comminuted fractures of the distal radius admitted to our hospital from March 2022 to February 2023.Among them,48 patients underwent wrist arthroscopy assisted closed reduction with external fixation with a bracket(arthroscopy group),and another 50 patients underwent open reduction with steel plate internal fixation surgery(control group).The two groups were compared in terms of operation time,intraoperative bleeding volume,incision length,fracture union time,range of motion(ROM)of joint,Visual Analogue Scale(VAS),and Gartland-Werley wrist scores,as well as radiographic parameters(palmar tilt,ulnar inclination,and radial height)evaluated at various follow-up intervals.Results The arthroscopy group had an operation time of(55.3±10.5)min,which was significantly shorter than that in the control group[(83.4±14.6)min;t=-10.979,P=0.000].The intraoperative bleeding volume in the arthroscopy group was(42.3±8.6)ml,which was less than that in the control group[(71.4±10.5)ml;t=-14.953,P=0.000].The incision length of the arthroscopy group was(1.3±0.3)cm,which was shorter than that of the control group[(5.1±1.5)cm;t=-18.550,P=0.000].The arthroscopy group had a fracture healing time of(10.7±1.4)weeks,which was shorter than that in the control group[(12.2±1.6)weeks;t=-4.855,P=0.000].The palmar flexion ROM in the arthroscopy group was 68.8°±8.3°,which was significantly higher than that in the control group(61.5°±9.4°;t=4.002,P=0.000).The dorsiflexion ROM in the arthroscopy group was 63.9°±7.5°,which was significantly higher than that in the comtrol group(59.2°±8.3°;t=2.931,P=0.004).The pronation ROM in the arthroscopy group was 67.4°±10.3°,which was significantly higher than that in the control group(62.1°±9.9°;t=2.604,P=0.011).The supination ROM in the arthroscopy group was 70.5°±7.4°,which was significantly higher than that in the control group(64.4°±8.6°;t=3.777,P=0.000).The VAS score of the arthroscopy group was(1.3±0.6)points,which was significantly lower than that in the control group[(1.7±0.5)points;t=-3.941,P=0.000].After 6 months,the Gartland-Werley wrist scores of the arthroscopy group was significantly higher than that of the control group(Z=-2.614,P=0.009).The wrist joint imaging showed significantly higher radial height,palmar inclination angle,and ulnar deviation angle in the arthroscopy group than the control group(all P=0.000),while there were no significant differences in palmar inclination angle and ulnar deviation angle at different time points within each group(P>0.05).Except for significant differences in radial height at 3 d,1 month,and 6 months after surgery(P=0.015,P=0.035),there were no significant differences between any other time points(P>0.05).The interaction between time and group was not significant for palm inclination angle,ulnar deviation angle,and radial height(P>0.05).Conclusion Wrist arthroscopy assisted closed reduction with external fixation for intra-articular comminuted fractures of distal radius has advantages of short operation time,less intraoperative blood loss,and good recovery of wrist joint functions.
2.Comparison of Wrist Arthroscopy Assisted Closed Reduction With External Fixation and Open Reduction With Steel Plate Internal Fixation for Intra-articular Comminuted Fractures of the Distal Radius
Zizhen LIU ; Guangxin CHEN ; Bin TANG ; Zhaoxin LUAN ; Zhengxun LI
Chinese Journal of Minimally Invasive Surgery 2025;25(9):550-556
Objective To explore the clinical efficacy of wrist arthroscopy assisted closed reduction with external fixation in the treatment of intra-articular comminuted fractures of the distal radius(type C3).Methods A retrospective analysis was conducted on 98 patients with type C3 intra-articular comminuted fractures of the distal radius admitted to our hospital from March 2022 to February 2023.Among them,48 patients underwent wrist arthroscopy assisted closed reduction with external fixation with a bracket(arthroscopy group),and another 50 patients underwent open reduction with steel plate internal fixation surgery(control group).The two groups were compared in terms of operation time,intraoperative bleeding volume,incision length,fracture union time,range of motion(ROM)of joint,Visual Analogue Scale(VAS),and Gartland-Werley wrist scores,as well as radiographic parameters(palmar tilt,ulnar inclination,and radial height)evaluated at various follow-up intervals.Results The arthroscopy group had an operation time of(55.3±10.5)min,which was significantly shorter than that in the control group[(83.4±14.6)min;t=-10.979,P=0.000].The intraoperative bleeding volume in the arthroscopy group was(42.3±8.6)ml,which was less than that in the control group[(71.4±10.5)ml;t=-14.953,P=0.000].The incision length of the arthroscopy group was(1.3±0.3)cm,which was shorter than that of the control group[(5.1±1.5)cm;t=-18.550,P=0.000].The arthroscopy group had a fracture healing time of(10.7±1.4)weeks,which was shorter than that in the control group[(12.2±1.6)weeks;t=-4.855,P=0.000].The palmar flexion ROM in the arthroscopy group was 68.8°±8.3°,which was significantly higher than that in the control group(61.5°±9.4°;t=4.002,P=0.000).The dorsiflexion ROM in the arthroscopy group was 63.9°±7.5°,which was significantly higher than that in the comtrol group(59.2°±8.3°;t=2.931,P=0.004).The pronation ROM in the arthroscopy group was 67.4°±10.3°,which was significantly higher than that in the control group(62.1°±9.9°;t=2.604,P=0.011).The supination ROM in the arthroscopy group was 70.5°±7.4°,which was significantly higher than that in the control group(64.4°±8.6°;t=3.777,P=0.000).The VAS score of the arthroscopy group was(1.3±0.6)points,which was significantly lower than that in the control group[(1.7±0.5)points;t=-3.941,P=0.000].After 6 months,the Gartland-Werley wrist scores of the arthroscopy group was significantly higher than that of the control group(Z=-2.614,P=0.009).The wrist joint imaging showed significantly higher radial height,palmar inclination angle,and ulnar deviation angle in the arthroscopy group than the control group(all P=0.000),while there were no significant differences in palmar inclination angle and ulnar deviation angle at different time points within each group(P>0.05).Except for significant differences in radial height at 3 d,1 month,and 6 months after surgery(P=0.015,P=0.035),there were no significant differences between any other time points(P>0.05).The interaction between time and group was not significant for palm inclination angle,ulnar deviation angle,and radial height(P>0.05).Conclusion Wrist arthroscopy assisted closed reduction with external fixation for intra-articular comminuted fractures of distal radius has advantages of short operation time,less intraoperative blood loss,and good recovery of wrist joint functions.
3.Effect of Modified Cangfu Daotantang on Glucose and Lipid Metabolism in Simple Obese Children with Phlegm Dampness and Stagnation
Zhenfang LUAN ; Fengru WANG ; Bin LUO ; Xiaochang WANG ; Tingting WANG ; Lizhen YANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(3):90-97
ObjectiveTo observe and compare the intervention effect of modified Cangfu Daotantang on glucose and lipid metabolism in simple obese children with phlegm dampness and stagnation. MethodA total of 60 children with simple obesity were randomly divided into two groups according to the simple randomization method of the random number table. The odd number was included in the test group, and the even number was included in the basic treatment group, with 30 cases in each group. On the basis of signing the informed consent notice, the treatment group was given modified Cangfu Daotantang combined with basic treatment, while the control group was only given basic treatment. After three months of treatment, the body mass index (BMI), glucose and lipid metabolism level [total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), fasting plasma glucose (FPG), fasting insulin (FINS), and homeostasis model assessment-insulin resistance (HOMA-IR)], the change in the total score of traditional Chinese medicine (TCM) syndromes, and the effective rate of treatment were observed and compared. ResultAfter treatment, the BMI of the observation group and the control group decreased significantly (P<0.01). Compared with the control group, the BMI level in the observation group decreased significantly (P<0.05). After treatment, the levels of TC, TG, and LDL-C in the observation group and the control group decreased significantly (P<0.01). Compared with the control group, the levels of TC, TG, and LDL-C in the observation group decreased significantly (P<0.05). In addition, the level of TC in the observation group improved significantly compared with that in the control group (P<0.01). The levels of FPG, FINS, and HOMA-IR in the observation group and the control group were significantly lower than those before treatment (P<0.05). After treatment, compared with the control group, the levels of FPG, FINS, and HOMA-IR in the observation group were significantly reduced (P<0.05). The level of FPG in the observation group was significantly improved compared with that in the control group (P<0.01). After treatment, the total score of TCM syndromes in the two groups decreased significantly (P<0.01). Compared with the control group, the total score of TCM syndromes in the observation group was lower (P<0.01). After treatment, the total effective rate of treatment was 86.67% (26/30) in the observation group and 73.33% (22/30) in the control group. By rank sum test, the total effective rate of the observation group was better than that of the control group (Z=-2.100, P<0.05). ConclusionModified Cangfu Daotantang combined with basic treatment can effectively reduce the BMI of obese children and improve their glucose and lipid metabolism. It has good clinical effects and high clinical application value, which is worth further in-depth research and promotion.
4.Preliminary evaluation of immunogenicity and protective effect of multicomponent recombinant protein vaccine EPRHP014 against tuberculosis
Bin CAO ; Xueting FAN ; Ruihuan WANG ; Xiuli LUAN ; Chengyu QIAN ; Jinjie YU ; Haican LIU ; Machao LI ; Guilian LI ; Xiuqin ZHAO ; Xiuqin YUAN ; Kanglin WAN
Chinese Journal of Epidemiology 2023;44(10):1653-1660
Objective:To evaluate the immunogenicity and protective effect of a multicomponent recombinant protein vaccine EPRHP014 constructed independently and provide a scientific basis for developing new tuberculosis (TB) vaccine and effective prevention and control of TB.Methods:Three full-length Mycobacterium ( M.) tuberculosis protein antigens (EsxH, Rv2628, and HspX) and two epitope-predicted and optimized epitope-dominant protein antigens (nPPE18 and nPstS1) were selected, from which five protein antigens were used to construct a protein antigen composition EPRHP014, including a fusion expression multi-component protein antigen (EPRHP014f) and a multi-component mixed protein antigen (EPRHP014m) formed with the five single protein using clone, purification, and purification respectively. Multicomponent protein vaccines EPRHP014f and EPRHP014m were prepared with aluminum adjuvant, and the BCG vaccine was used as a control. ELISA detected the titer of serum-specific antibodies, the secretion of various cytokines was detected by ELISpot and Luminex, and immune protection was observed by the M.tuberculosis growth inhibition test in vitro. The results were statistically analyzed by t-test or rank sum test, and P<0.05 was considered a statistically significant difference. Results:Mice Immunized with EPRHP014m and EPRHP014f could produce highly effective IgG antibodies and their subtypes IgG1 and IgG2a, and the antibody titers were similar to those of mice immunized with BCG, with no statistical significance ( P>0.05). The number of spot-forming cells (SFC) secreting IFN-γ and IL-4 induced by EPRHP014f group was significantly higher than those by EPRHP014m group and BCG group ( P<0.05), but there was no significant difference in the number of SFC for IFN-γ and IL-4 induced between EPRHP014m group and BCG group ( P>0.05). The secretion levels of GM-CSF and IL-12p70 induced by the EPRHP014m group were higher than those of the BCG group ( P<0.05), but there was no significant difference in the levels of IL-6 and IL-10 induced between EPRHP014m group and BCG group ( P>0.05). There was no significant difference in the secretions of IL-6, IL-10, IL-12, and GM-CSF between the EPRHP014f and BCG groups ( P>0.05). EPRHP014m group, EPRHP014f group, and BCG group had obvious antibacterial effects in vitro, and the difference was insignificant ( P>0.05). Conclusion:Both EPRHP014f and EPRHP014m can induce strong humoral and cellular immune responses in mice after immunization, and have a strong ability to inhibit the growth of M. tuberculosis in vitro, indicating that the antigen composition EPRHP014 has good potential in the development and application of TB vaccine.
5.Preliminary immunological evaluation of Mycobacterium tuberculosis multicomponent protein vaccine candidates EPDPA015f and EPDPA015m
Ruihuan WANG ; Xueting FAN ; Chengyu QIAN ; Bin CAO ; Jinjie YU ; Machao LI ; Guilian LI ; Xiuqin ZHAO ; Xiuli LUAN ; Haican LIU ; Kanglin WAN
Chinese Journal of Microbiology and Immunology 2023;43(4):294-303
Objective:To preliminarily evaluate the immunogenicity and efficacy of two novel tuberculosis vaccine candidates (a fusion multicomponent protein EPDPA015f and a mixed multicomponent protein EPDPA015m) and to provide a new antigen combination for the development of tuberculosis vaccines.Methods:Recombinant plasmids for the expression of EPDPA015f and EPDPA015m proteins were constructed. Six-week-old BALB/c mice were immunized with EPDPA015f or EPDPA015m in combination with aluminium adjuvant (50 μg/mouse) for three times with an interval of 10 d. The mice were sacrificed 10 d after the last immunization to collect blood and spleen samples. Serum antibody titers and cytokine levels were measured by ELISA, Luminex technique and enzyme-linked immunospot assay (ELISPOT). Mycobacterial growth inhibition assay (MGIA) was used to detect the ability of mouse splenocytes to inhibit the growth of Mtb in vitro. One-way analysis of variance and t-test were used for statistical analysis. Results:Both EPDPA015f and EPDPA015m could induce the production of various cytokines and IgG antibodies at a high level. The levels of cytokines related to Th1 (IL-2, TNF-α, IFN-γ), Th2 (IL-4, IL-6, IL-10) and Th17 (IL-17) as well as other proinflammatory cytokines (GM-CSF, IL-12) were higher in the EPDPA015f group than in the adjuvant group ( P<0.05). The titer of IgG antibody induced by EPDPA015f was as high as 1∶4×10 6. The results of MGIA showed that the numbers of Mtb (lgCFU) in the PBS, adjuvant, EPDPA015f and EPDPA015m groups were 3.46±0.11, 3.51±0.06, 2.98±0.09 and 3.19±0.08, respectively. The number of colonies in the EPDPA015f group was the least as compared with that in the other three groups ( P<0.001, P<0.001, P<0.01). Conclusions:The vaccine candidate EPDPA015f could elicit more comprehensive and high-level cellular and humoral immune responses, and exhibited superior in vitro inhibitory activity against the growth of Mtb. EPDPA015f had the potential to be used as a preventive vaccine or a booster vaccine
6.Spatiotemporal Dynamics of the Molecular Expression Pattern and Intercellular Interactions in the Glial Scar Response to Spinal Cord Injury.
Leilei GONG ; Yun GU ; Xiaoxiao HAN ; Chengcheng LUAN ; Chang LIU ; Xinghui WANG ; Yufeng SUN ; Mengru ZHENG ; Mengya FANG ; Shuhai YANG ; Lai XU ; Hualin SUN ; Bin YU ; Xiaosong GU ; Songlin ZHOU
Neuroscience Bulletin 2023;39(2):213-244
Nerve regeneration in adult mammalian spinal cord is poor because of the lack of intrinsic regeneration of neurons and extrinsic factors - the glial scar is triggered by injury and inhibits or promotes regeneration. Recent technological advances in spatial transcriptomics (ST) provide a unique opportunity to decipher most genes systematically throughout scar formation, which remains poorly understood. Here, we first constructed the tissue-wide gene expression patterns of mouse spinal cords over the course of scar formation using ST after spinal cord injury from 32 samples. Locally, we profiled gene expression gradients from the leading edge to the core of the scar areas to further understand the scar microenvironment, such as neurotransmitter disorders, activation of the pro-inflammatory response, neurotoxic saturated lipids, angiogenesis, obstructed axon extension, and extracellular structure re-organization. In addition, we described 21 cell transcriptional states during scar formation and delineated the origins, functional diversity, and possible trajectories of subpopulations of fibroblasts, glia, and immune cells. Specifically, we found some regulators in special cell types, such as Thbs1 and Col1a2 in macrophages, CD36 and Postn in fibroblasts, Plxnb2 and Nxpe3 in microglia, Clu in astrocytes, and CD74 in oligodendrocytes. Furthermore, salvianolic acid B, a blood-brain barrier permeation and CD36 inhibitor, was administered after surgery and found to remedy fibrosis. Subsequently, we described the extent of the scar boundary and profiled the bidirectional ligand-receptor interactions at the neighboring cluster boundary, contributing to maintain scar architecture during gliosis and fibrosis, and found that GPR37L1_PSAP, and GPR37_PSAP were the most significant gene-pairs among microglia, fibroblasts, and astrocytes. Last, we quantified the fraction of scar-resident cells and proposed four possible phases of scar formation: macrophage infiltration, proliferation and differentiation of scar-resident cells, scar emergence, and scar stationary. Together, these profiles delineated the spatial heterogeneity of the scar, confirmed the previous concepts about scar architecture, provided some new clues for scar formation, and served as a valuable resource for the treatment of central nervous system injury.
Mice
;
Animals
;
Gliosis/pathology*
;
Cicatrix/pathology*
;
Spinal Cord Injuries
;
Astrocytes/metabolism*
;
Spinal Cord/pathology*
;
Fibrosis
;
Mammals
;
Receptors, G-Protein-Coupled
7.Short-term visual quality observation of femtosecond laser small incision lenticule extraction and evolution implantable collamer lens implantation in the correction of moderate myopia
Hao-Ying YAO ; Jing-Lin LUAN ; Shao-Bin ZHANG ; Hao-Run ZHANG ; Rui WANG ; Jing-Jing ZHAO ; Meng-Jun FU
International Eye Science 2023;23(4):567-572
AIM: To investigate the short-term visual quality outcomes after femtosecond laser small incision lenticule extraction(SMILE)and evolution implantable collamer lens(EVO-ICL)implantation for the correction of moderate myopia.METHODS: Prospective control study. A total of 51 cases(51 eyes)with moderate myopia who underwent SMILE or EVO-ICL implantation surgery at Weifang Eye Hospital from April 2021 to February 2022 were selected. They were divided into SMILE group(30 patients, 30 eyes)and EVO-ICL group(21 patients, 21 eyes)according to the surgical methods. The changes of visual acuity [uncorrected distance visual acuity(UDVA), corrected distance visual acuity(CDVA)], diopter [spherical equivalent(SE)] and related parameters of optical quality analysis system(OQAS Ⅱ)were observed before surgery and at 1wk, 1 and 3mo after surgery, and the quality of vision(QoV)questionnaire was completed.RESULTS: At 3mo after surgery, the safety index(postoperative CDVA/preoperative CDVA)of SMILE gruop and EVO-ICL group were 1.20(1.00, 1.20)and 1.20(1.00, 1.38), respectively, the efficacy index(postoperative UDVA/preoperative CDVA)were 1.00(1.00, 1.20)and 1.00(1.00, 1.20), respectively, and the percentage of SE within ±0.50D was 87% and 100%, respectively. In SMILE group, the objective scattering index(OSI)was increased after surgery, while modulation transfer function cutoff frequency(MTF cutoff), contrast visual acuity(VA)100%, and VA20% at 1wk and 1mo after surgery, and Strehl ratio(SR)and VA9% at each time point after surgery were all decreased compared with those before surgery(all P<0.05). The OSI, MTF cutoff, SR and VA of EVO-ICL group showed no difference at each time point after surgery compared with those before surgery(all P>0.05). The most common visual symptoms after SMILE and EVO-ICL implantation were visual haze and halos, respectively.CONCLUSION: Both SMILE and EVO-ICL implantation have good safety, efficacy and predictability in the short term after the correction of moderate myopia. Both groups had visual symptoms after surgery, but the overall satisfaction of patients was high. Furthermore, EVO-ICL implantation has better objective visual quality performance.
8.Inhibitory effect and molecular mechanism of sinomenine on human hepatocellular carcinoma HepG2 and SK-HEP-1 cells.
Ying-Ying TIAN ; Bei-Bei MA ; Xin-Yue ZHAO ; Chuang LIU ; Yi-Lin LI ; Shang-Yue YU ; Shi-Qiu TIAN ; Hai-Luan PEI ; Ying-Nan LYU ; Ze-Ping ZUO ; Zhi-Bin WANG
China Journal of Chinese Materia Medica 2023;48(17):4702-4710
This study aimed to investigate the effect and molecular mechanism of sinomenine on proliferation, apoptosis, metastasis, and combination with inhibitors in human hepatocellular carcinoma HepG2 cells and SK-HEP-1 cells. The effect of sinomenine on the growth ability of HepG2 and SK-HEP-1 cells were investigated by CCK-8 assay, colony formation assay, and BeyoClick~(TM) EdU-488 staining. The effect of sinomenine on DNA damage was detected by immunofluorescence assay, and the effect of sinomenine on apoptosis of human hepatocellular carcinoma cells was clarified by Hoechst 33258 staining and CellEvent~(TM) Cystein-3/7Green ReadyProbes~(TM) reagent assay. Cell invasion assay and 3D tumor cell spheroid invasion assay were performed to investigate the effect of sinomenine on the invasion ability of human hepatocellular carcinoma cells in vitro. The effect of sinomenine on the regulation of protein expression related to the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription 3(STAT3) signaling pathway in HepG2 and SK-HEP-1 cells was examined by Western blot. Molecular docking was used to evaluate the strength of affinity of sinomenine to the target cysteinyl aspartate specific proteinase-3(caspase-3) and STAT3, and combined with CCK-8 assay to detect the changes in cell viability after combination with STAT3 inhibitor JSI-124 in combination with CCK-8 assay. The results showed that sinomenine could significantly reduce the cell viability of human hepatocellular carcinoma cells in a concentration-and time-dependent manner, significantly inhibit the clonogenic ability of human hepatocellular carcinoma cells, and weaken the invasive ability of human hepatocellular carcinoma cells in vitro. In addition, sinomenine could up-regulate the cleaved level of poly ADP-ribose polymerase(PARP), a marker of apoptosis, and down-regulate the protein levels of p-Akt, p-mTOR, and p-STAT3 in human hepatocellular carcinoma cells. Molecular docking results showed that sinomenine had good affinity with the targets caspase-3 and STAT3, and the sensitivity of sinomenine to hepatocellular carcinoma cells was diminished after STAT3 was inhibited. Therefore, sinomenine can inhibit the proliferation and invasion of human hepatocellular carcinoma cells and induce apoptosis, and the mechanism may be attributed to the activation of caspase-3 signaling and inhibition of the Akt/mTOR/STAT3 pathway. This study can provide a new reference for the in-depth research and clinical application of sinomenine and is of great significance to further promote the scientific development and utilization of sinomenine.
Humans
;
Carcinoma, Hepatocellular/genetics*
;
Proto-Oncogene Proteins c-akt/metabolism*
;
Caspase 3/metabolism*
;
Liver Neoplasms/genetics*
;
Molecular Docking Simulation
;
Sincalide/pharmacology*
;
Cell Line, Tumor
;
Cell Proliferation
;
Hep G2 Cells
;
TOR Serine-Threonine Kinases/metabolism*
;
Apoptosis
9.Morin induces autophagy and apoptosis in hepatocellular carcinoma cells through Akt/mTOR/STAT3 pathway.
Xin-Yue ZHAO ; Ying-Ying TIAN ; Chuang LIU ; Yi-Lin LI ; Ying-Nan LYU ; Shang-Yue YU ; Shi-Qiu TIAN ; Hai-Luan PEI ; Ze-Ping ZUO ; Zhi-Bin WANG
China Journal of Chinese Materia Medica 2023;48(16):4475-4482
This study investigated the effect and mechanism of morin in inducing autophagy and apoptosis in hepatocellular carcinoma cells through the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription protein 3(STAT3) pathway. Human hepatocellular carcinoma SK-HEP-1 cells were stimulated with different concentrations of morin(0, 50, 100, 125, 200, and 250 μmol·L~(-1)). The effect of morin on the viability of SK-HEP-1 cells was detected by Cell Counting Kit-8(CCK-8). The effect of morin on the proliferation and apoptosis of SK-HEP-1 cells was investigated using colony formation assay, flow cytometry, and BeyoClick~(TM) EdU-488 with different concentrations of morin(0, 125, and 250 μmol·L~(-1)). The changes in the autophagy level of cells treated with morin were examined by transmission electron microscopy and autophagy inhibitors. The impact of morin on the expression levels of proteins related to the Akt/mTOR/STAT3 pathway was verified by Western blot. Compared with the control group, the morin groups showed decreased viability of SK-HEP-1 cells in a time-and concentration-dependent manner, increased number of apoptotic cells, up-regulated expression level of apoptosis marker PARP, up-regulated phosphorylation level of apoptosis-regulating protein H2AX, decreased number of positive cells and the colony formation rate, an upward trend of expression levels of autophagy-related proteins LC3-Ⅱ, Atg5, and Atg7, and decreased phosphorylation levels of Akt, mTOR, and STAT3. These results suggest that morin can promote apoptosis, inhibit proliferation, and induce autophagy in hepatocellular carcinoma cells, and its mechanism of action may be related to the Akt/mTOR/STAT3 pathway.
Humans
;
Proto-Oncogene Proteins c-akt/metabolism*
;
Carcinoma, Hepatocellular/pathology*
;
Liver Neoplasms/pathology*
;
TOR Serine-Threonine Kinases/metabolism*
;
Apoptosis
;
Autophagy
;
Cell Proliferation
;
Cell Line, Tumor
;
STAT3 Transcription Factor/metabolism*
10.Scutellarin alleviates lipopolysaccharide-induced renal injury via mediating cysteine-rich protein 61-connective tissue growth factor-nephroblastoma overexpressed gene 1 expression to inhibit nuclear factor-κB signaling pathway
Xue LIU ; Zhenzhi QIN ; Chen GUAN ; Lingyu XU ; Jie DAI ; Chengyu YANG ; Bin ZHOU ; Hong LUAN ; Long ZHAO ; Wei ZHANG ; Congjuan LUO ; Yan XU
Chinese Critical Care Medicine 2022;34(4):400-406
Objective:To explore the protective effect and mechanism of scutellarin (Scu) on sepsis associated-acute kidney injury (SA-AKI).Methods:① In vivo experiment: 36 male C57BL/6 mice were divided into normal saline (NS) control group, lipopolysaccharide (LPS) induced SA-AKI model group (LPS group), 20 mg/kg Scu control group (Scu 20 control group), and 5, 10, 20 mg/kg Scu pretreatment groups by random number table with 6 mice in each group. The SA-AKI model was reproduced by intraperitoneal injection of 10 mg/kg LPS. The NS control group was injected with NS intraperitoneally. The Scu pretreatment groups were intraperitoneally injected with different doses of Scu every day before LPS injection for 1 week. Scu 20 control group was injected with 20 mg/kg Scu for 1 week. After 24 hours of LPS treatment, mice in each group were sacrificed, kidney tissues were collected, and kidney injury was detected by hematoxylin-eosin (HE) staining. Western blotting was used to detect the protein expression levels of nuclear factor-κB (NF-κB) signaling pathway related molecules, apoptosis-related proteins and cysteine-rich protein 61-connective tissue growth factor-nephroblastoma overexpressed gene 1 (CCN1). ② In vitro experiment: human renal tubular epithelial cell line HK-2 was cultured in vitro and used for experiment when the cells fused to 80%. In the cells without LPS treatment and after 100 g/L LPS treatment, pcDNA3.1-CCN1 and small interfering RNA (siRNA) CCN1 sequence were transfected to overexpress and inhibit CCN1 expression, respectively, to observe whether CCN1 was involved in NF-κB signaling pathway activation and apoptosis. In addition, 100g/L LPS and 20 μmol/L Scu were added into HK-2 cells transfected with and without CCN1 siRNA to investigate the mechanism of protective effect of Scu on LPS-induced HK-2 cells injury. Results:① The results of in vivo experiment: the renal function of SA-AKI mice induced by LPS was significantly decreased, and had kidney histological damage and severely damaged renal tubules. Scu could alleviate renal function and histological damage in a dose-dependent manner. Western blotting results showed Scu could reduce the protein expression of NF-κB signaling pathway related molecules and CCN1 in the renal tissue, and had a significant alleviating effect on apoptosis, indicating that CCN1 was involved in NF-κB signaling pathway activation and apoptosis. ② The results of in vitro experiment: in HK-2 cells not treated with LPS, CCN1 overexpression had no effect on apoptosis related protein and pro-inflammatory factors of NF-κB signaling pathway. In HK-2 cells treated with LPS, overexpression of CCN1 significantly inhibited the mRNA expressions of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCP-1), with significant differences as compared with cells stimulated only by LPS [IL-1β mRNA (2 -ΔΔCT): 3.20±0.57 vs. 4.88±0.69, TNF-α mRNA (2 -ΔΔCT): 2.99±0.44 vs. 5.00±0.81, MCP-1 mRNA (2 -ΔΔCT): 2.81±0.50 vs. 5.41±0.75, all P < 0.05], and the apoptosis-related protein was significantly down-regulated. However, when siRNA was used to inhibit the expression of CCN1, the mRNA expressions of pro-inflammatory factors were significantly increased as compared with cells stimulated only by LPS [IL-1β mRNA (2 -ΔΔCT): 6.01±1.13 vs. 4.88±0.69, TNF-α mRNA (2 -ΔΔCT): 5.15±0.86 vs. 5.00±0.81, all P < 0.05], and apoptosis-related protein was significantly up-regulated. In the LPS-induced HK-2 cells, the mRNA expressions of pro-inflammatory factors were significantly down-regulated after Scu treatment as compared with cells stimulated only by LPS [IL-1β mRNA (2 -ΔΔCT) : 2.55±0.50 vs. 6.15±1.04, TNF-α mRNA (2 -ΔΔCT): 2.58±0.40 vs. 3.95±0.52, MCP-1 mRNA (2 -ΔΔCT): 2.64±0.44 vs. 6.21±0.96, all P < 0.05], and apoptosis-related protein was also significantly reduced. When the expression of CCN1 was inhibited by siRNA, the protective effect of Scu on cells was weakened, which showed that the mRNA expressions of pro-inflammatory factors in cells was significantly up-regulated compared with the cells without inhibition of CCN1 expression [IL-1β mRNA (2 -ΔΔCT): 5.34±0.76 vs. 2.55±0.50, TNF-α mRNA (2 -ΔΔCT): 3.66±0.54 vs. 2.58±0.40, MCP-1 mRNA (2 -ΔΔCT): 5.15±0.79 vs. 2.64±0.44, all P < 0.05], and the expression of apoptosis related protein was also significantly up-regulated. Conclusions:Scu could protect the renal function in SA-AKI mice, and the protective effect is associated with NF-κB signaling pathway and CCN1. Thus, Scu could alleviate LPS-induced kidney injury by regulating the NF-κB signaling pathway.

Result Analysis
Print
Save
E-mail