Objective To investigate whether Porphyromonas gingivalis(Pg)in the oral cavity can influence the occurrence of perioperative neurocognitive disorder(PND)by modulating the TLR4/NF-κB signaling pathway.Methods In cell experi-ment,BV-2 cells in the logarithmic growth phase were randomly divided into two groups:the control group(Group C)and the LPS-Pg-treatedgroup(Group P).Western blot was used to detect the expression levels of signaling proteins(TLR4,NF-κB p65,NLRP3,TNF-α)in cellular proteins.Immunofluorescence assay was employed to observe thechanges in NF-κB p65 expres-sions inside and outside the nucleus.In animal experiments,C57BL/6 mice were randomly divided into six groups:the control group(Group C),the surgery group(Group S),the Pg infection group(Group P),the Pg infection+surgery group(Group PS),the TLR4inhibitor group(Group T),and the Pg infection+surgery+TLR4 inhibitor group(Group PST).After corresponding treatments,the Morris water maze(MWM)test was applied to observe theeffects of Pg infection on postoperativecognitive be-havior in mice.Immunohistochemical analysis was used to assess the impact of Pg infection on the activation of microglia in the hippocampal tissue of mice.Results In cell experiment,compared with Group C,expression levels of TLR4,NLRP3,and TNF-α increased significantly in Group P(all P＜0.05).The expression of NF-κB p65 in the nucleus increased(P＜0.05),while its expression outside the nucleus decreased(P＜0.05).Immunofluorescence results showed increased NF-κB expression and its translocation into the nucleus in Group P.As for animal experiment,no significant difference was observed in the preoperative training phase among the groups in Morris water maze experiment,with similar escape latencies and average swimming speeds(all P＞0.05).Therefore,therewas no notabledifferences in navigationand swimming abilities among thegroups before theex-periment.In the postoperative testing phase,compared with Group C,the target quadrant time and platform-crossing times were less in mice of Group S and Group PS(all P＜0.05).Compared with Group PS,the target quadrant time and platform-crossing times were elevated in mice of Group PST(both P＜0.05).Compared to Group C and Group PST,number of positive cells in hippocampal tissue was higher in Group PS.These positivecells exhibited enlarged somas,shortened processes,and an activated state of microglia.Conclusion Pg infection induces enhanced central nervous system inflammation during the perioperative peri-od in mice,leading to the occurrence of PND,possibly through the activation of the TLR4/NF-κB signaling pathway.

Objective To investigate the effects of Remimazolam combined with Esketamine on cellular immunity,cognitive function,and sleep quality in elderly patients with lung cancer during surgery.Methods A total of 106 elderly lung cancer patients who underwent elective thoracoscopic lobectomy from September 2023 to March 2025 were selected and divided into the control group(n=53)and the research group(n=53)according to random number table method.The control group received Midazolam combined with Esketamine for general anesthesia,while the research group received Remimazolam combined with Esketamine for general anesthesia.The intraoperative and postoperative conditions,hemodynamic parameters[heart rate(HR),mean arterial pressure(MAP)]before anesthesia induction(T0),at tracheal intubation(T1),at 1 min before one-lung ventilation(OLV)(T2),at skin incision(T3),at 1 h after OLV(T4),and immediately after extubation(T5),as well as pain and inflammatory factors[5-hydroxytryptamine(5-HT),prostaglandin E2(PGE2),high-sensitivity C-reactive protein(hs-CRP),tumor necrosis factor-α(TNF-α)],cellular immunity[peripheral blood natural killer(NK)cells,CD3+,CD4+,CD4+/CD8+],the levels of serum interleukin(IL)-2,interferon-γ(IFN-γ),cognitive function[Mini-Mental State Examination(MMSE)],sleep quality[Pittsburgh Sleep Quality Index(PSQI)],and recovery quality[Postoperative Quality of Recovery-15(QoR-15)score]were compared between the two groups of patients.The adverse reactions and complications were also compared between the two groups.Results There was no significant difference in duration of surgery,duration of anesthesia,intraoperative blood loss,and time to spontaneous breathing recovery of patients between the two groups(P>0.05).The total dosage of Sufentanil in the research group was less than that in the control group,and time to postoperative eye opening,duration of recovery room stay and length of postoperative hospitalization were shorter than those in the control group(P<0.05).At T0 and T5,there was no significant difference in HR and MAP between the two groups(P>0.05),and at T1-T4,HR and MAP in the research group were higher than those in the control group(P<0.05).At the time of recovery,the levels of serum 5-HT,PGE2,hs-CRP,and TNF-α in the research group were lower than those in the control group(P<0.05),and at the time of recovery,the peripheral blood NK cells,CD3+,CD4+,and CD4+/CD8+ratio,serum IL-2 and IFN-γ levels in the research group were higher than those in the control group(P<0.05).At 1 d after surgery,the MMSE and QoR-15 scores of the research group were higher than those of the control group,while the PSQI score was lower than that of the control group(P<0.05).The incidence of adverse reactions and complications in the research group[5.66%(3/53)and 3.77%(2/53)],was lower than that of the control group[18.87%(10/53)and 15.09%(8/53)](P<0.05).Conclusion The combination of Remimazolam and Esketamine in elderly patients undergoing thoracoscopic lobectomy can better maintain intraoperative hemodynamics,stabilize cellular immune function,reduce serum pain and inflammatory factor levels,improve postoperative cognitive function and sleep quality,reduce adverse reactions and complications,and promote postoperative recovery.

Objective To investigate whether Porphyromonas gingivalis(Pg)in the oral cavity can influence the occurrence of perioperative neurocognitive disorder(PND)by modulating the TLR4/NF-κB signaling pathway.Methods In cell experi-ment,BV-2 cells in the logarithmic growth phase were randomly divided into two groups:the control group(Group C)and the LPS-Pg-treatedgroup(Group P).Western blot was used to detect the expression levels of signaling proteins(TLR4,NF-κB p65,NLRP3,TNF-α)in cellular proteins.Immunofluorescence assay was employed to observe thechanges in NF-κB p65 expres-sions inside and outside the nucleus.In animal experiments,C57BL/6 mice were randomly divided into six groups:the control group(Group C),the surgery group(Group S),the Pg infection group(Group P),the Pg infection+surgery group(Group PS),the TLR4inhibitor group(Group T),and the Pg infection+surgery+TLR4 inhibitor group(Group PST).After corresponding treatments,the Morris water maze(MWM)test was applied to observe theeffects of Pg infection on postoperativecognitive be-havior in mice.Immunohistochemical analysis was used to assess the impact of Pg infection on the activation of microglia in the hippocampal tissue of mice.Results In cell experiment,compared with Group C,expression levels of TLR4,NLRP3,and TNF-α increased significantly in Group P(all P＜0.05).The expression of NF-κB p65 in the nucleus increased(P＜0.05),while its expression outside the nucleus decreased(P＜0.05).Immunofluorescence results showed increased NF-κB expression and its translocation into the nucleus in Group P.As for animal experiment,no significant difference was observed in the preoperative training phase among the groups in Morris water maze experiment,with similar escape latencies and average swimming speeds(all P＞0.05).Therefore,therewas no notabledifferences in navigationand swimming abilities among thegroups before theex-periment.In the postoperative testing phase,compared with Group C,the target quadrant time and platform-crossing times were less in mice of Group S and Group PS(all P＜0.05).Compared with Group PS,the target quadrant time and platform-crossing times were elevated in mice of Group PST(both P＜0.05).Compared to Group C and Group PST,number of positive cells in hippocampal tissue was higher in Group PS.These positivecells exhibited enlarged somas,shortened processes,and an activated state of microglia.Conclusion Pg infection induces enhanced central nervous system inflammation during the perioperative peri-od in mice,leading to the occurrence of PND,possibly through the activation of the TLR4/NF-κB signaling pathway.

Objective To investigate the effects of Remimazolam combined with Esketamine on cellular immunity,cognitive function,and sleep quality in elderly patients with lung cancer during surgery.Methods A total of 106 elderly lung cancer patients who underwent elective thoracoscopic lobectomy from September 2023 to March 2025 were selected and divided into the control group(n=53)and the research group(n=53)according to random number table method.The control group received Midazolam combined with Esketamine for general anesthesia,while the research group received Remimazolam combined with Esketamine for general anesthesia.The intraoperative and postoperative conditions,hemodynamic parameters[heart rate(HR),mean arterial pressure(MAP)]before anesthesia induction(T0),at tracheal intubation(T1),at 1 min before one-lung ventilation(OLV)(T2),at skin incision(T3),at 1 h after OLV(T4),and immediately after extubation(T5),as well as pain and inflammatory factors[5-hydroxytryptamine(5-HT),prostaglandin E2(PGE2),high-sensitivity C-reactive protein(hs-CRP),tumor necrosis factor-α(TNF-α)],cellular immunity[peripheral blood natural killer(NK)cells,CD3+,CD4+,CD4+/CD8+],the levels of serum interleukin(IL)-2,interferon-γ(IFN-γ),cognitive function[Mini-Mental State Examination(MMSE)],sleep quality[Pittsburgh Sleep Quality Index(PSQI)],and recovery quality[Postoperative Quality of Recovery-15(QoR-15)score]were compared between the two groups of patients.The adverse reactions and complications were also compared between the two groups.Results There was no significant difference in duration of surgery,duration of anesthesia,intraoperative blood loss,and time to spontaneous breathing recovery of patients between the two groups(P>0.05).The total dosage of Sufentanil in the research group was less than that in the control group,and time to postoperative eye opening,duration of recovery room stay and length of postoperative hospitalization were shorter than those in the control group(P<0.05).At T0 and T5,there was no significant difference in HR and MAP between the two groups(P>0.05),and at T1-T4,HR and MAP in the research group were higher than those in the control group(P<0.05).At the time of recovery,the levels of serum 5-HT,PGE2,hs-CRP,and TNF-α in the research group were lower than those in the control group(P<0.05),and at the time of recovery,the peripheral blood NK cells,CD3+,CD4+,and CD4+/CD8+ratio,serum IL-2 and IFN-γ levels in the research group were higher than those in the control group(P<0.05).At 1 d after surgery,the MMSE and QoR-15 scores of the research group were higher than those of the control group,while the PSQI score was lower than that of the control group(P<0.05).The incidence of adverse reactions and complications in the research group[5.66%(3/53)and 3.77%(2/53)],was lower than that of the control group[18.87%(10/53)and 15.09%(8/53)](P<0.05).Conclusion The combination of Remimazolam and Esketamine in elderly patients undergoing thoracoscopic lobectomy can better maintain intraoperative hemodynamics,stabilize cellular immune function,reduce serum pain and inflammatory factor levels,improve postoperative cognitive function and sleep quality,reduce adverse reactions and complications,and promote postoperative recovery.

Objective:To explore the effects and underlying mechanism of human adipose mesenchymal stem cells (ADSC)-derived exosomes on acute lung injury in septic mice.Methods:The study was an experimental study. Human ADSC of passages 4-5 were selected, and exosomes in their supernatant were isolated and extracted by differential ultracentrifugation. Exosomes were then used after identification. Twenty-four adult male BALB/c mice were selected and divided into normal control group, simple cecal ligation and puncture (CLP) group, and CLP+ADSC-exosome group according to the random number table method (the grouping method was the same below), with 8 mice in each group. The mice in simple CLP group were injected with phosphate buffer after CLP surgery (to establish an animal model of acute lung injury in septic mice), the mice in CLP+ADSC-exosome group were treated according to the corresponding group name, and the mice in normal control group were only injected with phosphate buffer. At 24 hours after surgery, the morphology of lung tissue was observed by hematoxylin-eosin staining, the apoptosis of lung tissue cells was detected by in-situ end-labeling method, the content of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the serum of mice was detected by enzyme-linked immunosorbent assay, the content of malondialdehyde and superoxide dismutase (SOD) in lung tissue was detected by microplate reader, and the expressions of CD86 and CD206 in mouse lung tissue cells was detected by immunofluorescence method. Mouse macrophage RAW264.7 was taken and divided into blank control group, simple lipopolysaccharide (LPS) group, and LPS+ADSC-exosome group. The cells of LPS+ADSC-exosome group and simple LPS group were cultured by adding LPS+ADSC-exosome and LPS, respectively, and cells in blank control group were routinely cultured. Twelve hours after culture, the ATP content, the percentage of mitochondrial reactive oxygen species positive cells, as well as mitochondrial membrane potential in cells were detected by related detection kits. The mRNA expression levels of M1 polarization marker inducible nitric oxide synthase (iNOS), M2 polarization marker arginase-1 (Arg1), and inflammatory factors TNF-α and IL-1β in cells were detected by real-time fluorescence quantitative reverse-transcription polymerase chain reaction method. Three samples were used for mRNA expression detection, and four samples were used for the detection of the other indicators.Results:At 24 hours after surgery, the structure of mouse lung tissues in normal control group was clear and intact without inflammatory cell infiltration. Compared with that in normal control group, the lung tissue edema as well as the infiltration of inflammatory cells of mice was much more obvious in simple CLP group. However, compared with that in simple CLP group, the lung tissue edema of mice in CLP+ADSC-exosome group was significantly alleviated, the infiltration of inflammatory cells was significantly reduced, and the cell apoptosis and necrosis were significantly improved. Twenty-four hours after surgery, compared with that in normal control group, the levels of TNF-α and IL-1β in the serum of mice in simple CLP group were significantly increased (with t values of 50.82 and 30.81, respectively, P<0.05); compared with that in simple CLP group, the levels of TNF-α and IL-1β in the serum of mice in CLP+ADSC-exosome group were significantly decreased (with t values of 16.36 and 19.25, respectively, P<0.05). Compared with that in normal control group, the content of malondialdehyde in the lung tissue of mice in simple CLP group was significantly increased ( t=9.89, P<0.05); and the content of SOD was significantly decreased ( t=5.01, P<0.05); compared with that in simple CLP group, the content of malondialdehyde in the lung tissue of mice in CLP+ADSC-exosome group was significantly decreased ( t=4.38, P<0.05), and the content of SOD was significantly increased ( t=2.97, P<0.05). Twenty-four hours after surgery, compared with that in normal control group, the proportion of CD86 positive cells in the lung tissue of mice in simple CLP group was significantly increased, and the proportion of CD206 positive cells was significantly decreased; compared with that in simple CLP group, the proportion of CD86 positive cells in the lung tissue of mice in CLP+ADSC-exosome group was significantly decreased, and the proportion of CD206 positive cells was significantly increased. After 12 hours of culture, compared with that in blank control group, the ATP content of RAW264.7 cells in simple LPS group was significantly decreased ( t=6.28, P<0.05); compared with that in simple LPS group, the ATP content of RAW264.7 cells in LPS+ADSC-exosome group was significantly increased ( t=4.01, P<0.05). After 12 hours of culture, compared with (22±4)% in blank control group, (40±6)% of positive cells of mitochondrial reactive oxygen species in RAW264.7 cells in simple LPS group was significantly increased ( t=5.04, P<0.05); compared with that in LPS group, (30±5)% of positive cells of mitochondrial reactive oxygen species in RAW264.7 cells in LPS+ADSC-exosome group was significantly decreased ( t=2.65, P<0.05). After 12 hours of culture, compared with that in blank control group, the mitochondrial membrane potential of RAW264.7 cells in simple LPS group was significantly decreased; the mitochondrial membrane potential of RAW264.7 cells in LPS+ ADSC-exosome group was between those in blank control group and simple LPS group. After 12 hours of culture, compared with that in blank control group, the mRNA expressions of TNF-α, IL-1β, and iNOS in RAW264.7 cells in simple LPS group were significantly increased (with t values of 16.51, 31.04, and 7.70, respectively, P<0.05), and the decrease in the mRNA expression of Arg1 was not statistically significant ( P>0.05); compared with that in simple LPS group, the mRNA expressions of TNF-α, IL-1β, and iNOS in RAW264.7 cells in LPS+ADSC-exosome group were significantly decreased (with t values of 11.38, 22.58, and 5.28, respectively, P<0.05), and the mRNA expression of Arg1 was significantly increased ( t=7.66, P<0.05). Conclusions:Human ADSC-exosomes may play a role in improving lung injury in septic mice by improving LPS-induced mitochondrial dysfunction in mice macrophages, inhibiting the polarization of macrophages toward M1, and reducing the inflammatory response.

Objective:To explore the effects and underlying mechanism of human adipose mesenchymal stem cells (ADSC)-derived exosomes on acute lung injury in septic mice.Methods:The study was an experimental study. Human ADSC of passages 4-5 were selected, and exosomes in their supernatant were isolated and extracted by differential ultracentrifugation. Exosomes were then used after identification. Twenty-four adult male BALB/c mice were selected and divided into normal control group, simple cecal ligation and puncture (CLP) group, and CLP+ADSC-exosome group according to the random number table method (the grouping method was the same below), with 8 mice in each group. The mice in simple CLP group were injected with phosphate buffer after CLP surgery (to establish an animal model of acute lung injury in septic mice), the mice in CLP+ADSC-exosome group were treated according to the corresponding group name, and the mice in normal control group were only injected with phosphate buffer. At 24 hours after surgery, the morphology of lung tissue was observed by hematoxylin-eosin staining, the apoptosis of lung tissue cells was detected by in-situ end-labeling method, the content of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the serum of mice was detected by enzyme-linked immunosorbent assay, the content of malondialdehyde and superoxide dismutase (SOD) in lung tissue was detected by microplate reader, and the expressions of CD86 and CD206 in mouse lung tissue cells was detected by immunofluorescence method. Mouse macrophage RAW264.7 was taken and divided into blank control group, simple lipopolysaccharide (LPS) group, and LPS+ADSC-exosome group. The cells of LPS+ADSC-exosome group and simple LPS group were cultured by adding LPS+ADSC-exosome and LPS, respectively, and cells in blank control group were routinely cultured. Twelve hours after culture, the ATP content, the percentage of mitochondrial reactive oxygen species positive cells, as well as mitochondrial membrane potential in cells were detected by related detection kits. The mRNA expression levels of M1 polarization marker inducible nitric oxide synthase (iNOS), M2 polarization marker arginase-1 (Arg1), and inflammatory factors TNF-α and IL-1β in cells were detected by real-time fluorescence quantitative reverse-transcription polymerase chain reaction method. Three samples were used for mRNA expression detection, and four samples were used for the detection of the other indicators.Results:At 24 hours after surgery, the structure of mouse lung tissues in normal control group was clear and intact without inflammatory cell infiltration. Compared with that in normal control group, the lung tissue edema as well as the infiltration of inflammatory cells of mice was much more obvious in simple CLP group. However, compared with that in simple CLP group, the lung tissue edema of mice in CLP+ADSC-exosome group was significantly alleviated, the infiltration of inflammatory cells was significantly reduced, and the cell apoptosis and necrosis were significantly improved. Twenty-four hours after surgery, compared with that in normal control group, the levels of TNF-α and IL-1β in the serum of mice in simple CLP group were significantly increased (with t values of 50.82 and 30.81, respectively, P<0.05); compared with that in simple CLP group, the levels of TNF-α and IL-1β in the serum of mice in CLP+ADSC-exosome group were significantly decreased (with t values of 16.36 and 19.25, respectively, P<0.05). Compared with that in normal control group, the content of malondialdehyde in the lung tissue of mice in simple CLP group was significantly increased ( t=9.89, P<0.05); and the content of SOD was significantly decreased ( t=5.01, P<0.05); compared with that in simple CLP group, the content of malondialdehyde in the lung tissue of mice in CLP+ADSC-exosome group was significantly decreased ( t=4.38, P<0.05), and the content of SOD was significantly increased ( t=2.97, P<0.05). Twenty-four hours after surgery, compared with that in normal control group, the proportion of CD86 positive cells in the lung tissue of mice in simple CLP group was significantly increased, and the proportion of CD206 positive cells was significantly decreased; compared with that in simple CLP group, the proportion of CD86 positive cells in the lung tissue of mice in CLP+ADSC-exosome group was significantly decreased, and the proportion of CD206 positive cells was significantly increased. After 12 hours of culture, compared with that in blank control group, the ATP content of RAW264.7 cells in simple LPS group was significantly decreased ( t=6.28, P<0.05); compared with that in simple LPS group, the ATP content of RAW264.7 cells in LPS+ADSC-exosome group was significantly increased ( t=4.01, P<0.05). After 12 hours of culture, compared with (22±4)% in blank control group, (40±6)% of positive cells of mitochondrial reactive oxygen species in RAW264.7 cells in simple LPS group was significantly increased ( t=5.04, P<0.05); compared with that in LPS group, (30±5)% of positive cells of mitochondrial reactive oxygen species in RAW264.7 cells in LPS+ADSC-exosome group was significantly decreased ( t=2.65, P<0.05). After 12 hours of culture, compared with that in blank control group, the mitochondrial membrane potential of RAW264.7 cells in simple LPS group was significantly decreased; the mitochondrial membrane potential of RAW264.7 cells in LPS+ ADSC-exosome group was between those in blank control group and simple LPS group. After 12 hours of culture, compared with that in blank control group, the mRNA expressions of TNF-α, IL-1β, and iNOS in RAW264.7 cells in simple LPS group were significantly increased (with t values of 16.51, 31.04, and 7.70, respectively, P<0.05), and the decrease in the mRNA expression of Arg1 was not statistically significant ( P>0.05); compared with that in simple LPS group, the mRNA expressions of TNF-α, IL-1β, and iNOS in RAW264.7 cells in LPS+ADSC-exosome group were significantly decreased (with t values of 11.38, 22.58, and 5.28, respectively, P<0.05), and the mRNA expression of Arg1 was significantly increased ( t=7.66, P<0.05). Conclusions:Human ADSC-exosomes may play a role in improving lung injury in septic mice by improving LPS-induced mitochondrial dysfunction in mice macrophages, inhibiting the polarization of macrophages toward M1, and reducing the inflammatory response.

Objective:To observe the effects of pyrroloquinoline quinine (PQQ) on the mitochondrial function and cell survival of rat bone marrow mesenchymal stem cells (BMSCs) under oxidative stress, and to explore its mechanism.Methods:BMSCs of rats were cultured in vitro with Dulbecco′s minimum essential medium/F12 medium containing fetal bovine serum in the volume fraction of 10% (hereinafter referred to as normal medium). The rat BMSCs of third to fifth passages in logarithmic growth phase were selected for the following experiments. (1) The cells were divided into normal control group, normal control+ PQQ group, hydrogen peroxide (H 2O 2) alone group, and H 2O 2+ PQQ group. The cells in normal control group were cultured in normal medium for 24 hours; the cells in normal control+ PQQ group were cultured in normal medium containing 100 μmol/L PQQ for 24 hours; the cells in H 2O 2 alone group were cultured in normal medium containing 200 μmol/L H 2O 2 for 24 hours; the cells in H 2O 2+ PQQ group were pre-incubated with normal medium containing 100 μmol/L PQQ for 2 hours, and then with H 2O 2 added to the concentration of 200 μmol/L and cultured for 24 hours. The cell morphology of each group was observed under the inverted phase contrast microscope, and the cell survival rate was detected by cell count kit 8 method. (2) Five batches of cells were collected, and the cells of each batch were divided into normal control group, H 2O 2 alone group, and H 2O 2+ PQQ group. The cells in each group received the same treatment as that in the corresponding group of experiment (1). After 24 hours of culture, one batch of cells was collected for apoptosis detection by flow cytometry, and the apoptosis rate was calculated. One batch of cells was subjected to mitochondrial membrane potential assay and JC-1 fluorescent staining observation using the JC-1 mitochondrial membrane potential detection kit and the inverted phase contrast fluorescence microscope, respectively. One batch of cells was collected for mitochondrial morphology observation under the transmission electron microscope. One batch of cells was subjected to catalase (CAT) and superoxide dismutase (SOD) activity assay by CAT activity assay kit and SOD activity assay kit, respectively. One batch of cells was subjected to Western blotting for determination of protein level of Epac1, adenine monophosphate activated protein kinase (AMPK), phosphorylated AMPK, cysteinyl aspartate-specific proteinase 3 (caspase-3), and cleaved caspase-3, and the phosphorylation level of AMPK and cleaved caspase-3/caspase-3 ratio were calculated. Six replicates were measured in each group for each index except for morphological observation. Data were statistically analyzed with one-way analysis of variance and independent sample equal variance t test. Results:(1) After 24 hours of culture, compared with those in normal control group (the cell survival rate was set to 100.0%), there was an increase in cell vacuole and a decrease in cell number in H 2O 2 alone group, and the cell survival rate was significantly reduced to (74.3±2.9)% ( t=6.39, P<0.01). Compared with those in H 2O 2 alone group, the cell morphology of H 2O 2+ PQQ group was significantly improved, and the cell survival rate was significantly increased to (116.9±4.2)% ( t=6.92, P<0.01); the cell survival rate in normal control+ PQQ group was (101.2±1.1)%, close to that of control group ( t=1.06, P>0.05). (2) After 24 hours of culture, compared with (13.6±1.0)% in normal control group, the apoptosis rate of cells in H 2O 2 alone group was significantly increased to (37.1±2.0)% ( t=10.57, P<0.01). Compared with that in H 2O 2 alone group, the apoptosis rate of cells in H 2O 2+ PQQ group was significantly declined to (17.0±0.7)% ( t=9.49, P<0.01). (3) After 24 hours of culture, compared with those in normal control group, the mitochondrial membrane potential of cells in H 2O 2 alone group was depolarized, the JC-1 fluorescent dye mainly existed in the cytoplasm in the form of monomer, which emitted green fluorescence, and a significant decrease in mitochondrial membrane potential was shown ( t=4.18, P<0.01). Compared with those in H 2O 2 alone group, the mitochondrial membrane potential of cells in H 2O 2+ PQQ group was increased to normal level ( t=4.43, P<0.01), and the JC-1 fluorescent dye accumulated in mitochondria following the polarized mitochondrial membrane potential and emitted red fluorescence. (4) After 24 hours of culture, compared with that in normal control group, the mitochondrial structure of cells in H 2O 2 alone group was disordered, with disappeared mitochondrial cristae and decreased mitochondrial matrix density. Compared with that in H 2O 2 alone group, the mitochondrial structure of cells in H 2O 2+ PQQ group was regular and intact, with clearly visible mitochondrial cristae and increased mitochondrial matrix density. (5) After 24 hours of culture, compared with those in normal control group, the CAT activity of cells in H 2O 2 alone group was significantly increased ( t=4.54, P<0.05), and the SOD activity was significantly decreased ( t=3.93, P<0.05). Compared with those in H 2O 2 alone group, the CAT activity of cells in H 2O 2+ PQQ group was obviously increased ( t=8.65, P<0.01), while there was no significant change in the SOD activity ( t=0.72, P>0.05). (6) After 24 hours of culture, compared with those in normal control group, the protein expression of Epac1 of cells in H 2O 2 alone group was significantly decreased ( t=4.67, P<0.01), while the AMPK phosphorylation level and the cleaved caspase-3/caspase-3 ratio were significantly increased ( t=7.88, 3.62, P<0.01). Compared with those in H 2O 2 alone group, the protein expression of Epac1 and the AMPK phosphorylation level of cells in H 2O 2+ PQQ group were both significantly increased ( t=4.34, 16.37, P<0.01), while the cleaved caspase-3/caspase-3 ratio was significantly declined ( t=3.17, P<0.05). Conclusions:Pretreatment with PQQ can improve the mitochondrial function, reduce cell apoptosis rate, and enhance cell survival rate of rat BMSCs under oxidative stress, which may be related to the up-regulation of Epac1 protein expression, activation of AMPK signaling pathway, and down-regulation of cleaved caspase-3 protein level.

Objective: To explore the effects of microRNA-34a on regulating silent information regulator 1 (SIRT1) and influence of SIRT1 on myocardial damage of rats with severe burns at early stage. Methods: (1) Twenty-four Sprague-Dawley (SD) rats were divided into sham injury (SI) group, simple burns (SB) group and SIRT1 agonist (SA) group according to the random number table (the same grouping method below), with 8 rats in each group. Rats in groups SB and SA were inflicted with 30% total body surface area full-thickness scald (hereinafter referred to as burns) on the back, and rats in group SI were sham injuried on the back. Immediately after injury, rats in groups SI and SB were intraperitoneally injected with normal saline of 50 mL/kg, and rats in group SA were intraperitoneally injected with normal saline of 50 mL/kg and 1 mg/mL resveratrol of 50 mg/kg. At 6 h post injury, abdominal aortic blood was collected to make serum and myocardial tissue of rats was collected. (2) Myocardial cells of twelve neonatal SD rats were collected and divided into microRNA-34a mimic control (MMC) group, microRNA-34a mimic (MM) group, microRNA-34a inhibitor control (MIC) group, and microRNA-34a inhibitor (MI) group, which were respectively transfected with gene sequences of mimic control, mimic, inhibitor control, and inhibitor of microRNA-34a. The microRNA-34a expression level and protein expression level of SIRT1 in myocardial cells were respectively detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. Another batch of myocardial cells were divided into microRNA-34a inhibitor control+ burn serum (MCB) group, microRNA-34a inhibitor+ burn serum (MB) group, and microRNA-34a inhibitor+ burn serum + EX527 (MBE) group. Myocardial cells in group MCB were transfected with gene sequence of inhibitor control, and myocardial cells in the later groups were transfected with gene sequence of inhibitor of microRNA-34a. After transfection of 48 h, myocardial cells in group MBE were cultured in Dulbecco′s modified Eagle′s medium (DMEM) solution for 6 hours, with serum in group SB of volume fraction of 10% and final amount-of-substance concentration of 1 mol/L, and myocardial cells in the other 2 groups were cultured in DMEM solution with serum from rats of group SB of volume fraction of 10%. The protein expression levels of myocardial cells of SIRT1, cleaved-caspase-3, and Bax were detected by Western blotting. (3) Myocardial tissue from (1) was collected to detect expression levels of microRNA-34a and mRNA of SIRT1 in groups SI and SB by real-time fluorescence quantitative RT-PCR. Morphology of myocardial tissue of rats in groups SI, SB, and SA was observed with biological image navigator. The mRNA expression levels of interleukin 1β (IL-1β) and tumor necrosis factor (TNF-α) of rats in groups SI, SB, and SA were detected by real-time fluorescence quantitative RT-PCR. The expression levels of cleaved-caspase-3, and Bax of myocardial tissue of rats in groups SI, SB, and SA were detected by Western blotting. Data were processed with one-way analysis of variance and least-significant difference test. Results: (1) After transfection of 48 h, the expression level of microRNA-34a of myocardial cells in group MM was 4.67±0.92, significantly higher than 1.03±0.04 in group MMC (P<0.01); the protein expression level of SIRT1 of myocardial cells in group MM was 0.35±0.06, significantly lower than 1.12±0.11 in group MMC (P<0.01). After transfection of 48 h, the expression level of microRNA-34a of myocardial cells in group MI was 0.26±0.07, significantly lower than 1.33±0.07 in group MIC (P<0.01); the protein expression level of SIRT1 of myocardial cells in group MIC was 1.12±0.16, significantly lower than 1.74±0.34 in group MI (P<0.01). At 6 h after culture, compared with those in group MCB, the SIRT1 protein expression level of myocardial cells in group MB was significantly increased (P<0.05), while cleaved-caspase-3 and Bax protein expression levels of myocardial cells in group MB were significantly decreased (P<0.05). Compared with those in group MB, the SIRT1 protein expression level of myocardial cells in group MBE was with no significantly statistical difference (P>0.05), and cleaved-caspase-3 and Bax protein expression levels were significantly increased (P<0.05). (2) At 6 h post injury, compared with that in group SI, the microRNA-34a expression level of myocardial tissue in group SB was significantly increased (P<0.01), and the mRNA expression level of SIRT1 of myocardial tissue in group SB was significantly decreased (P<0.01). At 6 h post injury, myocardial cells in group SI arranged neatly with normal nucleus and no inflammatory cells infiltration; myocardial cells in group SB arranged disorderly, with no abnormal nucleus, and obvious inflammatory cells infiltration; myocardial cells in group SA arranged neatly, with normal nucleus and little inflammatory cells infiltration. At 6 h post injury, compared with those in group SB, the mRNA expression levels of IL-1β and TNF-α, and the protein expression levels of cleaved-caspase-3 and Bax of myocardial tissue in groups SI and SA were significantly decreased (P<0.01). Conclusions The microRNA-34a expression level of myocardial tissue of rats with severe burns at early stage increases, which decreases the expression level of SIRT1, and increases the expression levels of IL-1β, TNF-α, cleaved-caspase-3 and Bax, leading to obvious myocardial damage. Activation of SIRT1 can alleviate myocardial damage of rats with severe burns at early stage through decreasing expression levels of IL-1β, TNF-α, cleaved-caspase-3, and Bax.

Objective To analyze the clinical efficacy and relative influence factors of self-expanding metallic stents (SEMS) in treating colorectal cancer obstruction.Methods Information of 47 patients received SEMS to treat colorectal cancer obstruction from Mar.2012 to Dec.2017 in Beijing Haidian Hospital were collected,who were then followed up in outpatient or by telephone.Effective rate,survival rate and complications were recorded and calculated by the software of SPSS 17.0.Chi-square test was used to analyze relative influence factors.Results Results shown that the clinical efficacy was 100％ (47/47).Patients' 30-day survival rate and 6-month survival rate were 87.2％ (41/47) and 68.1％ (32/47),respectively.After surgery,2 patients presented with perforation,3 patients presented with migration and 5 patients presented with reobstruction.Clinical stage of tumor and stent length were related with complications and survival.Therein,complications presented more in patients with advanced cancer.And the longer stent length,the higher mortality.Conclusions Clinical stage of tumor and stent length maybe risk factors of complications and survival of patients after SEMS surgery of colorectal cancer obstruction.

Objective: To investigate the effects of activating silent information regulator 1 (SIRT1) on the early kidney damage in rats with severe burn. Methods: Thirty healthy male SD rats were divided into sham injury group (SI), pure burn group (PB), and SIRT1 activator group (SA) according to the random number table, with 10 rats in each group. Rats in groups PB and SA were inflicted with 30% total body surface area full-thickness scald (hereinafter referred to as burn) on the back. Immediately after injury, rats in group PB were intraperitoneally injected with normal saline in the dosage of 50 mL/kg, and those in group SA with 1 mg/mL (final mass concentration) resveratrol in the dosage of 50 mL/kg. Rats in group SI were sham injured and intraperitoneally injected with normal saline in the dosage of 50 mL/kg immediately after injury. Kidney tissue and abdominal aorta blood of rats in the three groups were collected at 24 hours after injury. The morphology of kidney tissue was observed after HE staining. The serum content of creatinine and urea nitrogen was determined with enzyme-linked immunosorbent assay. Protein expressions of SIRT1, Bax, and Bcl-2 in kidney tissue were determined with Western blotting. mRNA expressions of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and IL-10 in kidney tissue were determined with real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were processed with one-way analysis of variance and LSD-t test. Results: (1) In rats of group SI, structures of kidney tubules and glomeruli were intact. In rats of group PB, structures of kidney tubules were not clear with casts in them, and glomeruli showed pyknosis. In rats of group SA, structures of kidney tubules were relatively intact, and the pyknosis of glomeruli were slighter as compared with that of group PB with fewer glomeruli showing pyknosis. (2) The serum content of creatinine and urea nitrogen in rats of group PB was (67±14) μmol/L and (22.0±4.4) mmol/L, respectively, which was significantly higher than that of group SI [(28±7) μmol/L and (5.5±1.2) mmol/L respectively, with t values respectively 6.07 and 11.53, P values below 0.01]. The serum content of creatinine and urea nitrogen in rats of group SA was (39±9) μmol/L and (14.1±1.7) mmol/L, respectively, significantly lower than that of group PB (with t values respectively 4.09 and 4.17, P values below 0.01). (3) Compared with those of group SI, protein expressions of SIRT1 and Bcl-2 in kidney tissue of rats in group PB were significantly decreased (with t values respectively 16.32 and 19.58, P values below 0.01), while the protein expression of Bax was significantly increased (t=5.98, P<0.01). Compared with those of group PB, protein expressions of SIRT1 and Bcl-2 in kidney tissue of rats in group SA were significantly increased (with t values respectively 6.94 and 5.37, P values below 0.01), while the protein expression of Bax was significantly decreased (t=3.44, P<0.01). (4) mRNA expressions of TNF-α, IL-1β, and IL-10 in kidney tissue of rats in group PB were 17.0±4.0, 2.27±0.59, and 2.5±0.9, respectively, significantly higher than those of group SI (1.0, 1.00, and 1.0, respectively, with t values from 3.27 to 8.93, P<0.05 or P<0.01). mRNA expressions of TNF-α and IL-1β in kidney tissue of rats in group SA were 6.8±1.2 and 1.18±0.26, respectively, significantly lower than those of group PB (with t values respectively 4.59 and 4.32, P values below 0.01). mRNA expression of IL-10 in kidney tissue of rats in group SA was 5.0±1.0, significantly higher than that of group PB (t=5.51, P<0.01). Conclusions Activating SIRT1 on early stage of severe burn in rats can decrease levels of creatinine and urea nitrogen, thus improving the kidney function. It can down-regulate the protein expression of Bax and up-regulate the protein expression of Bcl-2, thus reducing the apoptosis in kidney tissue. Meanwhile, it can inhibit expressions of TNF-α and IL-1β and promote the expression of IL-10, thus alleviating the inflammatory response in kidney.