The relation between left atrial (LA) size and myocardial pathological changes, and clinical implications were studied in 25 patients with- rheumatic mitral stenosis. The results showed that LA size was significantly correlated with pathological severity of myocardium (P 100 ml/m2 and accompanied by moderate or severe pathological changes easily suffered from Af (P200ml/m2 and severe pathological changes. The results suggest that occurence of Af may be predicated based on the LA volume and Af cardioversion should be selectively performed to obtain better results.

Objective: To investigate whether CD137 signaling can promote angiogenesis via regulating macrophage M1/M2 polarization. Methods: (1) The primary peritoneal macrophages in mice induced by 3% thiglycollate broth were divided into three groups: control group, CD137 signaling activated group and CD137 signaling inhibited group. Various specific markers of M1 and M2 macrophages were detected to observe the phenotype change of macrophages, and the macrophages protein expression of CD137, CD86 and CD206 was detected by flow cytometry (FCM). The protein and mRNA expression of induced nitric oxide synthase (iNOS), arginase Ⅰ(Arg-1) was determined by Western blot and RT-PCR, respectively. The secretion levels of IL-12 and IL-10 in culture supernatant of macrophages were detected by ELISA. (2) Macrophages were co-cultured with the endothelial cells (bEnd.3), and macrophages were implanted in the upper chamber, endothelial cells were implanted in stromal glue of the lower chamber. The experiment was divided into three groups: the control group, CD137 signaling activated group and peroxisome proliferator-activated receptor-γ (PPAR-γ) inhibited group, and tube formation ability of endothelial cells in each group was determined. Results: (1) The purity of primary peritoneal macrophages in mice was (97.93±1.31)%. The expression of CD137 on the surface of macrophages was (97.40±2.70)%. (2) Compared with control group, the mRNA and protein expression levels of Arg-1 were significantly increased and the mRNA and protein expression of iNOS were significantly decreased in CD137 signaling activated group (all P<0.05). Compared with CD137 signaling activated group, the mRNA and protein expression of Arg-1 were significantly lower and the mRNA and protein expression levels of iNOS were significantly higher in CD137 signaling inhibited group (all P<0.05). FCM results showed that the average fluorescence intensity of CD206 was higher, while the average fluorescence intensity of CD86 was lower in CD137 signaling activated group than in control group (P<0.05, P<0.01, respectively); the expression of CD206 was significantly lower, while the expression of CD86 was higher, in the CD137 signaling inhibited group than in CD137 signaling activated group (P<0.05, P<0.01, respectively). ELISA results showed that the secretion of IL-10 was higher, and the secretion level of IL-12 was significantly lower in CD137 signaling activated group than in control group (both P<0.01); the secretion of IL-10 was significantly lower and the secretion of IL-12 was significantly higher in CD137 signaling inhibited group than in CD137 signaling activated group (both P<0.05). (3) Values of the formation of tube length and branch number were both longer in CD137 signaling activated group than control group (P<0.05). The formation of the tube length and branch number were less in PPAR-γ inhibited group than in CD137 signaling activated group (P<0.05). Conclusion: CD137 signaling can promote angiogenesis by regulating macrophage M1/M2 polarization.
Animals ; Coculture Techniques ; Endothelial Cells ; Macrophages ; Mice ; Neovascularization, Pathologic ; Signal Transduction

OBJECTIVETo explore the inhibitive effects of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on expression of tissue factor (TF) in sepsis.

METHODSHuman umbilical vein endothelial cells (HUVEC) were cultured with trypsin digestion method, which were divided into NC group (with normal treatment), LPS group (with culture of 10 microg/mL LPS), LD group (with 10 microg/mL LPS and DMSO in co-culture), LC1 group (with 10 microg/mL LPS and 10 micromol/L CORM-2 in co-culture), LC2 group (with 10 microg/mL LPS and 50 micromol/L CORM-2 in co-culture), LC3 group (with 10 microg/mL LPS and 100 micromol/L CORM-2 in co-culture). After culture for 4 hours, TF activity, TF protein expression, nuclear factor-kappaB (NF-kappaB) activity were examined. Forty-five C57 BL/6 male mice were randomly divided into NC (without treatment, n = 5), CLP (n = 5) and CLP + CORM-2 (with treatment of 8 mg/kg CROM-2 after CLP, n = 5) groups. The serum samples in CLP, CLP + CORM-2 groups were collected at 2, 6, 12 and 24 post operation hour ( POH, 5 mice at each time point) for determination of TF and TFPI levels,which were also examined in NC group.

RESULTSCompared with those of NC group, TF activity increased (P < 0.01) , TF protein expression and NF-KB activity also increased in LPS group. Compared with those of LPS group, above indices were decreased in LC1, LC2, LC3 groups. The serum level of TF in CLP group at 6 POH was higher than that of NC group (80.0 +/- 11.9 pg/mL vs 58.4 +/- 6.9 pg/mL, P < 0.05), peaked at 12 POH, and still higher than that of NC group at 24 POH, while the serum level of TFPI showed no obvious difference in NC and CLP groups. Compared with that of NC group, TFPI levels obviously increased in CLP + CORM-2 group at 6, 12 POH (23.7 +/-3.5 ng/mL, 24.4 +/- 5.0 ng/mL vs 12.4 +/- 2.8 ng/mL, P < 0.05).

CONCLUSIONSExogenous CORM can obviously inhibit TF and NF-KB activity,decrease TF protein expression. Meanwhile, it can also decrease serum level of TF, and increase serum level of TFPI, preventing activation of procoagulant system, balancing procoagulant and anticoagulant system in sepsis.

Animals ; Cells, Cultured ; Humans ; Lipoproteins ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; metabolism ; Organometallic Compounds ; pharmacology ; Sepsis ; metabolism ; Thromboplastin ; metabolism

OBJECTIVETo study the inhibitive effect of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on the activation of Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway in sepsis.

METHODSRAW264.7 cells were divided into normal control group, LPS group (10 mg/mL LPS, the same concentration below), LPS + inactive CORM-2 (iCORM-2) group, LPS + 50 mmol/L CORM-2 group, and LPS + 100 mmol/L CORM-2 group. TNF-alpha level in the supernatant was determined with ELISA, and the phosphorylation levels of JAK1 and JAK3 were determined with Western blot. Thirty-five male BALB/c mice were divided into normal control group, cecal ligation and puncture (CLP) group, CLP + iCORM-2 (8.0 mg/kg) group and CLP + CORM-2 group (8.0 mg/kg) according to the random number table. Mice in CLP + CORM-2 group were treated the same as mice in CLP group except for administration of CORM-2 after CLP. The plasma levels of TNF-alpha, IL-1beta, and the phosphorylation levels of JAK1, JAK3 in liver tissue were determined with ELISA 24 hours post CLP. Data were processed with t test.

RESULTSCompared with that of normal control group [(1.9 +/- 0.3) pg/mL], the TNF-alpha level [(8.2 +/- 2.7) pg/mL, t = 2.844, P < 0.01] and phosphorylation levels of JAK1, JAK3 in LPS group increased significantly; while TNF-alpha levels in LPS + 50 mmol/L CORM-2 and LPS + 100 mmol/L CORM-2 groups decreased obviously as compared with that of LPS group [(5.7 +/- 1.4), (3.2 +/- 0.9) pg/mL, with t value respectively 2.104 and 2.363, P values all below 0.05], and it was the same with phosphorylation levels of JAK1, JAK3 in a dose-dependent manner. Compared with those of normal control group, plasma levels of TNF-alpha and IL-1beta and phosphorylation levels of JAK1, JAK3 in liver tissue significantly increased in CLP group (with t value respectively 2.916 and 2.796, and P values all below 0.05); while plasma levels of TNF-alpha and IL-1beta and the phosphorylation levels of JAK1, JAK3 in liver tissue decreased significantly in CLP + CORM-2 group (with t value respectively 2.115 and 2.398, and P values all below 0.05).

CONCLUSIONSExogenous CORM-2 can obviously inhibit the phosphorylation of JAKs molecules and then inhibit the activation of JAK/STAT signal pathway in sepsis, and decrease the expression of downstream cytokines to effectively prevent cascade reaction in the inflammatory response after severe infection.

Animals ; Carbon Monoxide ; pharmacology ; Cells, Cultured ; Interleukin-1beta ; blood ; Janus Kinase 1 ; metabolism ; Janus Kinase 3 ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Organometallic Compounds ; pharmacology ; Phosphorylation ; Sepsis ; metabolism ; Signal Transduction ; Tumor Necrosis Factor-alpha ; blood

Objective: To investigate whether CD137-CD137L signaling could affect the secretion of mouse vascular smooth muscle cells (VSMCs) -derived exosomes through autophagy mediated Rab7 pathway. Methods: Primary thoracic aorta VSMCs from C57BL/6J mouse were obtained by tissue block adherence method. VSMCs between the third to fifth passages were used and VSMCs were divided into 4 groups: control group, CD137 agonist group, lentivirus control group, Rab7 lentiviral interference group. VSMCs in CD137 agonist group were treated with recombinant protein of CD137L (10 μg/ml), VSMCs in lentivirus control group were treated with lentiviral followed by recombinant protein of CD137L (10 μg/ml), VSMCs in Rab7 lentiviral interference group were treated with Rab7 lentiviral intervention followed by recombinant protein of CD137L (10 μg/ml). Western blot was used to detect the protein expression of LC3Ⅱ, p62, Rab7, CD9, CD81 and Hsc70. Fluorescence microscopy was used to track the changes of autophagy in cells infected with mRFP-GFP-LC3. Transmission electron microscope was used to observe the morphology and size of VSMCs-derived exosomes. The nanoparticle tracking analysis(NTA) was used to detect the concentration and size of exosomes in each group. Results: (1) The expressions of Rab7, LC3Ⅱ and p62 protein in VSMCs of CD137 activation group were significantly higher than those in control group (all P<0.05). The expressions of Rab7, LC3Ⅱ and p62 protein in Rab7 lentivirus interference group was lower than in CD137 activation group (all P<0.05), while the expressions were similar between the lentivirus control group and the CD137 activation group (all P>0.05). (2) The total number of fluorescent spots and yellow fluorescent spots in the VSMCs of the CD137 activation group were higher than those in the control group (all P<0.05), and the number of yellow fluorescent spots was higher than that of the red fluorescent spots in the VSMCs of the CD137 activation group ((50.3±0.9) vs. (10.3±1.5)/cell). The total numbers of fluorescent spots and yellow fluorescent spots in VSMCs of Rab7 lentivirus interference group were lower than those of CD137 activation group (both P<0.05), and the number of red fluorescent spots in VSMCs was higher than that of yellow fluorescent spots ((40.7±4.0) and (10.7±1.2)/cell) in the Rab7 lentiviral interference group. The total numbers of fluorescent spots and yellow fluorescent spots in the VSMCs were similar between the lentivirus control group and the CD137 activation group (all P>0.05). (3) Under transmission electron microscopy, the size of the VSMCs-derived exosomes was about 30-150 nm. The exosome markers (CD9, CD81) could be detected in vesicles by Western blot. NTA results showed that the concentration of VSMCs-derived exosomes was significantly higher in the CD137-activated group than in the control group (P<0.05), which was significantly lower in the Rab7 lentiviral interference group than in the CD137-activation group (P<0.05) and was similar between the lentivirus control group and the CD137 activation group (P>0.05). The expression of Hsc70 protein in exosomes secreted by CD137 activation group was higher than that in the control group (P<0.05). The expression of Hsc70 protein in exosomes was lower in Rab7 lentivirus interference group than in the CD137 activation group (P<0.05), which was similar between the lentivirus control group and the CD137 activation group (P>0.05). The expression of LC3Ⅱ protein in exosome was higher in CD137 activation group than in control group (P<0.05), which was lower in Rab7 lentivirus interference group than in CD137 activation group (P<0.05), which was similar between the lentivirus control group and the CD137 activation group (P>0.05). Conclusion The CD137-CD137L signaling may affect the secretion of mouse VSMCs-derived exosomes through modulating the Rab7 pathway mediated autophagy.

OBJECTIVE: To investigate the mechanism of the in vitro toxicity of doxycycline to myeloma cell line H929 and also the possible pathway involved its toxicity. METHODS: Myeloma cell line H929 was treated with DOX, MEK inhibitor U0126 or RAS agonist ML-098, either alone or in combination. Then, the expression of p-MEK, caspase-3, caspase-9 and c-Jun in H929 were used to detected by Western blot; the cells proliferation and apoptosis were detected by CCK-8 assay and flow cytometry, respectively. RESULTS: DOX significantly increased the levels of cleaved caspase-3 and caspase-9, and down-regulated the level of p-MEK in H929 (P<0.05). MEK antagonist U0126 significantly increased the levels of cleaved caspase-3 and caspase-9, and down-regulated the level of p-MEK (P<0.05). After Dox combined with ML-098 treatment of H929 cells, the apoptosis rate of H929 cells was lower than that of DOX alone treatment group(P<0.05). Compared with DOX alone treatment group, the expressions of p-MEK and p-ERK1/2 in DOX+ML-098 combined treatment group were increased, and the levels of cleaved caspase-3,9 in H929 cells were decreased (P<0.05). The levels of c-Jun mRNA and protein increased in H929 when treated by DOX alone (P<0.05). CONCLUSION DOX can induce apoptosis of H929 via intrinsic apoptosis pathway, and MEK/ERK pathway and c-Jun possibly play a role in this process.
Apoptosis ; Caspase 3 ; Caspase 9/pharmacology* ; Cell Line, Tumor ; Cell Proliferation ; Doxycycline/pharmacology* ; Humans ; Mitogen-Activated Protein Kinase Kinases/pharmacology* ; Multiple Myeloma

Asiatic acid (AA), a pentacyclic triterpene found in, displays significant anti-proliferative effects on cancer cellsalthough the underlying mechanism of this effect remains unknown. This study investigated the efficacy and mechanism of action of AA against lung cancer bothand. Using the MTT assay, AA was found to induce apoptosis in a dose- and time-dependent manner, an effect enhanced by pretreatment with an autophagy inhibitor. It also elevated expression of microtubule-associated protein 1 light chain 3 (LC3) and decreased the expression of p62. Furthermore, exposure to AA resulted in collapse of the mitochondrial membrane potential and generation of reactive oxygen species (ROS), suggesting mitochondria are the target of AA. In the mouse lung cancer xenograft model, oral administration of AA significantly inhibited tumor volume and weight accompanied by significant apoptosis of lung cancer cells. In addition, it led to a significant decrease in the expression of proliferating cell nuclear antigen (PCNA). In summary, the results show that AA significantly reduces lung cancer cell growth bothandand that the associated apoptosis is mediated through mitochondrial damage.

Objective To investigate the probability and dosimetric risk factors of lung injury after sequential immune checkpoint inhibitors (ICIs) and thoracic radiotherapy. Methods A retrospective analysis was conducted on 139 patients who received sequential ICIs and thoracic radiotherapy in Xuzhou Cancer Hospital and Affiliated Hospital of Xuzhou Medical University between February 2020 and February 2024. The relationships of clinical factors and lung and heart volume dose parameters with grade ≥ 2 acute lung injury (ALI) in patients with thoracic tumors were studied using univariable (χ² test, t test, nonparametric test) and multivariable (binary logistic regression analysis) methods. The thresholds of dosimetric risk factors were determined using the receiver operating characteristic curves. Clinical factors included age, gender, smoking history, type of ICIs, cycle of ICI application, and the interval between ICI application and thoracic radiotherapy. Dose parameters included total radiotherapy dose, single dose, planning target volume, maximum dose of planning target volume, average dose of planning target volume, total lung volume, heart volume, and the V₅, V₁₀, V₁₅, V₂₀, V₂₅, V₃₀, V₃₅, and V₄₀ of lung and heart. Results The incidence of grade ≥ 2 ALI in the included cases was 36% (50/139). The χ² test did not find any statistically significant clinical factors. In the univariable and binary Logistic regression analysis, lung V₁₅ and V₂₀, heart V₁₅ and V₂₀, and lung volume were independent risk factors for the occurrence of grade ≥ 2 ALI in sequential ICIs and thoracic radiotherapy. The thresholds were 18.51% for lung V₁₅, 14.43% for lung V₂₀, 32.41% for heart V₁₅, and 17.74% for heart V₂₀. Conclusion For patients who are going to receive thoracic radiotherapy after ICIs, the thresholds of lung V₁₅ and V₂₀ and heart V₁₅ and V₂₀ in the radiotherapy plan are recommended to be less than 18.51%, 14.43%, 32.41%, and 17.74%, respectively, which can effectively reduce the occurrence of grade ≥ 2 ALI.

Exosomes are nanometer-sized vesicles that contain various types of biologically active components, including proteins, nucleic acids, carbohydrates, and lipids, which vary with the type and physiological state of the cell. In recent years, several studies have showed that exosomes can provide new non-invasive diagnostic and prognostic biomarkers in patients affected by cancers, including bladder cancer (BC), and the lipid bilayer membrane structure makes exosomes as promising delivery vehicles for therapeutic applications. Exosomes have the characteristics of high abundance, high stability, tissue specificity, and wide distribution in body fluids, and are secreted as various types by cells in different states, thereby possessing great potential as biomarkers for BC. Herein, we briefly summarize the functions and roles of exosomes in the occurrence and development of BC and the current progress of research on exosomes in BC, while focusing on potential clinical applications of the diagnosis, treatment, and prognosis of BC.

OBJECTIVE: To explore the bidirectional regulation of acupuncture based on a subgroup analysis of multicenter randomized controlled trial of acupuncture with METHODS: A total of 519 patients were included in the analysis, including 137 patients with constipation type irritable bowel syndrome (IBS-C) (92 cases in the acupuncture group and 45 cases in the polyethylene glycol [PEG] group), and 382 patients with diarrhea type irritable bowel syndrome (IBS-D) (252 cases in the acupuncture group and 130 cases in the pinaverium group). The patients in the acupuncture group were given acupuncture at Baihui (GV 20), Yintang (GV 29), Tianshu (ST 25), Shangjuxu (ST 37), Zusanli (ST 36), Sanyinjiao (SP 6) and Taichong (LR 3) once every other day, 3 times a week. The patients in the PEG group received polyethylene glycol 4000 powder orally, and the pinaverium group received pinaverium bromide tablets orally. All were treated for 6 weeks. The IBS symptom severity score (IBS-SSS) was assessed at baseline, treatment period (2, 4, 6 weeks of treatment) and 12 weeks of follow-up, and the IBS quality of life (IBS-QOL) score was evaluated at the baseline period, 6 weeks of treatment and 12 weeks of follow-up. RESULTS: The total IBS-SSS scores of the two groups of IBS-C patients at 2, 4, 6 weeks of treatment and follow-up of 12 weeks were lower than those in the baseline period ( CONCLUSION Acupuncture with
Acupuncture Points ; Acupuncture Therapy ; Diarrhea ; Humans ; Irritable Bowel Syndrome/therapy* ; Quality of Life ; Treatment Outcome