Role of HSF1 in endogenous protective mechanism underlying mechanical ventilator-induced lung injury in mice: relationship with HMGB1 / 中华麻醉学杂志

Objective:To evaluate the role of heat shock transcription factor 1 (HSF1) in the endogenous protective mechanism underlying mechanical ventilator-induced lung injury (VILI) in mice and the relationship with high mobility group box 1 (HMGB1).Methods:Forty SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=10 each) by the random number table method: control group (group C), VILI group (group VILI), negative control siRNA + VILI group (group NV) and HSF1 siRNA + VILI group (group siRNA). At 48 h before mechanical ventilation, negative control siRNA 5 nmol and HSF1 siRNA 5 nmol were intratracheally injected in NV and siRNA groups respectively, and the solution was diluted to 50 μl with the sterile phosphate buffer in both groups. Group C kept spontaneous breathing for 4 h, and the rest animals were mechanically ventilated (tidal volume 35 ml/kg, respiratory rate 75 breaths/min, inspiratory/expiratory ratio 1∶2, fraction of inspired oxygen 21%) for 4 h. Blood samples from the femoral artery were collected for arterial blood gas analysis immediately after endotracheal intubation and at 4 h of ventilation, and PaO 2 was recorded. Then the mice were sacrificed under deep anesthesia to collect lung tissues and bronchoalveolar lavage fluid (BALF). The concentrations of interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α) and HMGB1 in BALF were determined by enzyme-linked immunosorbent assay. The pathological results were observed by hematoxylin-eosin staining, and lung injury was assessed and scored. The wet/dry (W/D) weight ratio of lung tissues was calculated. The expression of HMGB1 and HSF1 mRNA in lung tissues (by quantitative real-time polymerase chain reaction) and expression of HMGB1 and HSF1 protein in lung tissues (by Western blot) were determined. Results:Compared with group C, PaO 2 was significantly decreased at 4 h of ventilation, the concentrations of TNF-α, IL-1β and HMGB1 in BALF, W/D ratio and lung injury score were increased, and the expression of HMGB1 protein and mRNA in lung tissues was up-regulated in group VILI, group NV and group siRNA ( P<0.05 or 0.01). Compared with group VILI and group NV, PaO 2 was significantly decreased at 4 h of ventilation, the concentrations of TNF-α, IL-1β and HMGB1 in BALF, W/D ratio and lung injury score were increased, and the expression of HMGB1 protein and mRNA in lung tissues was up-regulated, and the expression of HSF1 protein and mRNA was down-regulated in group siRNA ( P<0.05 or 0.01). There was no significant difference in the parameters mentioned above between group VILI and group NV ( P>0.05). Conclusions:HSF1 is involved in the endogenous protective mechanism underlying VILI in mice, which may be related to the down-regulation of HMGB1 expression and attenuation of inflammatory responses in lung tissues.

Changes of high mobility group protein B1 in condyloma acuminatum and its clinical significance / 中国医师进修杂志

Objective:To investigate the changes and clinical significance of high mobility group protein B1 (HMGB1) in condyloma acuminatum (CA).Methods:Sixty four patients with initial CA(initial group) and 48 patients with recurrent CA(recurrent goup) treated in the Second Affiliated Hospital of PLA Air Force Military Medical University Hospital from January 2019 to November 2020 were included. In the same period, 31 patients who underwent circumcision and 19 female underwent sexual organ cosmetic plastic surgery were taken as the control group, and the normal foreskin and healthy vulva were collected. The expression of HMGB1 in wart was detected by real-time quantitative polymerase chain reaction(RT-PCR), and the expression of soluble apoptosis related factor ligand (sFasL), cell lymphoma-2 gene (Bcl-2), soluble apoptosis related factor (SFAS) and Survivin, caspase-3 were detected. At the same time, serum interleukin (IL) - 6, IL-17, IL-23 and tumor necrosis factor - α (TNF- α) were detected by enzyme-linked immunosorbent assay(ELISA).Results:The expression of HMGB1 mRNA in the warts of patients in the initial group, recurrence group and control group were 1.96 ± 0.20, 1.53 ± 0.14, 1.05 ± 0.11, there was statistical difference ( F = 15.20, P<0.05) ; the expression of HMGB1 mRNA in the warts of patients in the initial group was significantly higher than that in the recurrence group and the control group ( P<0.05), and the recurrence group was also significantly higher than that in the control group ( P<0.05). The mRNA expressions of sFas, Bcl-2, sFasL and caspase-3 in the warts of patients in the initial group were significantly lower than those in the recurrence group and the control group: 0.52 ± 0.08 vs. 0.82 ± 0.16, 1.10 ± 0.19; 0.50 ± 0.05 vs. 0.79 ± 0.13, 1.08 ± 0.21; 0.47 ± 0.06 vs. 0.81 ± 0.15, 1.01 ± 0.19; 0.35 ± 0.04 vs. 0.68 ± 0.09, 0.91 ± 0.16, P<0.05; and the recurrence group were also significantly lower than those in the control group ( P<0.05). The expression of Survivin mRNA in the warts of patients in the initial group was significantly higher than those in the recurrence group and the control group: 2.14 ± 0.40 vs. 1.60 ± 0.27, 0.99 ± 0.18, P<0.05, and the recurrence group was also significantly higher than that in the control group ( P<0.05). The serum levels of TNF-α and IL-6 in the initial group were significantly lower than that in the recurrence group and the control group: (20.08 ± 1.95) μg/L vs. (26.93 ± 2.74), (37.65 ± 3.83) μg/L; (31.05 ± 3.24) μg/L vs. (38.13 ± 3.76), (45.98 ± 4.69) μg/L, P<0.05; and the recurrence group were also significantly lower than those in the control group ( P<0.05). The serum levels of IL-17 and IL-23 in the primary group were significantly higher than those in the recurrence group and the control group: (423.71 ± 28.68) ng/L vs. (384.26 ± 21.70) and (335.43 ± 19.65) ng/L; (289.50 ± 18.53) ng/L vs. (251.07 ± 15.96) and (214.67 ± 13.20) ng/L, P<0.05; and the recurrence group were also significantly higher than those in the control group ( P<0.05). The results of correlation analysis showed that the mRNA expression of HMGB1 in the warts of CA patients were negatively correlated with the mRNA expression of caspase-3, sFas, Bcl-2 and sFasL in the warts ( r = - 0.602, - 0.734, - 0.692, - 0.717, P<0.05), and was positive correlation with Survivin mRNA expression ( r = 0.645, P<0.05); and were positive correlation with the contents of IL-17 and IL-23 in serum ( r = 0.673, 0.685, P<0.05), and negatively correlated with the contents of TNF-α and IL-6 ( r = - 0.698, - 0.764, P<0.05). Conclusions:HMGB1 is obviously abnormal in the warts of patients with condyloma acuminatum, and is closely related to apoptosis, immune and inflammation-related factors, and may be jointly involved in the occurrence and recurrence of CA.

Hmga2 protein loss alters nuclear envelope and 3D chromatin structure.

BACKGROUND: The high-mobility group Hmga family of proteins are non-histone chromatin-interacting proteins which have been associated with a number of nuclear functions, including heterochromatin formation, replication, recombination, DNA repair, transcription, and formation of enhanceosomes. Due to its role based on dynamic interaction with chromatin, Hmga2 has a pathogenic role in diverse tumors and has been mainly studied in a cancer context; however, whether Hmga2 has similar physiological functions in normal cells remains less explored. Hmga2 was additionally shown to be required during the exit of embryonic stem cells (ESCs) from the ground state of pluripotency, to allow their transition into epiblast-like cells (EpiLCs), and here, we use that system to gain further understanding of normal Hmga2 function. RESULTS: We demonstrated that Hmga2 KO pluripotent stem cells fail to develop into EpiLCs. By using this experimental system, we studied the chromatin changes that take place upon the induction of EpiLCs and we observed that the loss of Hmga2 affects the histone mark H3K27me3, whose levels are higher in Hmga2 KO cells. Accordingly, a sustained expression of polycomb repressive complex 2 (PRC2), responsible for H3K27me3 deposition, was observed in KO cells. However, gene expression differences between differentiating wt vs Hmga2 KO cells did not show any significant enrichments of PRC2 targets. Similarly, endogenous Hmga2 association to chromatin in epiblast stem cells did not show any clear relationships with gene expression modification observed in Hmga2 KO. Hmga2 ChIP-seq confirmed that this protein preferentially binds to the chromatin regions associated with nuclear lamina. Starting from this observation, we demonstrated that nuclear lamina underwent severe alterations when Hmga2 KO or KD cells were induced to exit from the naïve state and this phenomenon is accompanied by a mislocalization of the heterochromatin mark H3K9me3 within the nucleus. As nuclear lamina (NL) is involved in the organization of 3D chromatin structure, we explored the possible effects of Hmga2 loss on this phenomenon. The analysis of Hi-C data in wt and Hmga2 KO cells allowed us to observe that inter-TAD (topologically associated domains) interactions in Hmga2 KO cells are different from those observed in wt cells. These differences clearly show a peculiar compartmentalization of inter-TAD interactions in chromatin regions associated or not to nuclear lamina. CONCLUSIONS: Overall, our results indicate that Hmga2 interacts with heterochromatic lamin-associated domains, and highlight a role for Hmga2 in the crosstalk between chromatin and nuclear lamina, affecting the establishment of inter-TAD interactions.

miR-142-3p simultaneously targets HMGA1, HMGA2, HMGB1, and HMGB3 and inhibits tumorigenic properties and in-vivo metastatic potential of human cervical cancer cells.

AIMS: High-mobility group (HMG) proteins are oncogenic in different cancers, including cervical cancer; silencing their individual expression using sh-RNAs, siRNAs, and miRNAs has had anti-tumorigenic effects, but the consequences of their collective downregulation are not known. Since multiple gene targeting is generally very effective in cancer therapy, the present study highlighted the consequences of silencing the expression of HMGA1, A2, B1, and B3 using sh-RNAs or miR-142-3p (that can potentially target HMGA1, A2, B1, and B3) in cervical cancer cell lines. MAIN METHODS: 3' UTR luciferase reporter assays were performed to validate HMGA1, A2, B1, and B3 as targets of miR-142-3p in human cervical cancer cells. Annexin V/PI dual staining and flow cytometry analyses were used to detect apoptotic cells. miR-142-3p-mediated regulation of cell death, colony formation, migration, and invasion was investigated in human cervical cancer cells together with in vivo metastasis in zebrafish. KEY FINDINGS: Concurrent knockdown of HMGA1, A2, B1, and B3 through their corresponding sh-RNAs inhibited cell viability and colony formation but induced apoptosis, and these effects were relatively reduced upon their individual knockdown. miR-142-3p targeted HMGA1, A2, B1, and B3 by binding to their 3'UTRs and induced apoptosis but inhibited proliferation, migration, and invasion of human cervical cancer cells. In addition, miR-142-3p expression decreased phospho-p65 and EMT-related proteins in cervical cancer cells and their in vivo metastatic potential upon implantation in zebrafish. SIGNIFICANCE: These findings suggest that miR-142-3p acts as a tumor-suppressive miRNA by targeting HMGA1, A2, B1, and B3 and may serve as a potential therapeutic agent in human cervical cancer.

Effect of lidocaine on expression of high mobility group box 1 protein mRNA during lidocaine-induced attenuation of pulmonary vascular endothelial dysfunction in septic rats / 中华麻醉学杂志

Objective:To evaluate the effect of lidocaine on the expression of high-mobility group box 1 protein (HMGB1) mRNA during lidocaine-induced attenuation of pulmonary vascular endothelial dysfunction in septic rats.Methods:Thirty clean-grade healthy male Sprague-Dawley rats, aged 8-12 weeks, weighing 250-300 g, were divided into 3 groups ( n=10 each) by the random number table method: sham operation group (S group), sepsis group (C group) and lidocaine group (L group). Sepsis model was developed by cecal ligation and puncture in anesthetized animals.In group S, only surgery was performed without ligation.In group L, lidocaine 10 mg/kg was injected into the tail vein immediately after model preparation, followed by 10 mg·kg -1·h -1 infusion for 3 h. Groups S and C received the equal volume of normal saline instead.At 24 h after model preparation, rats were sacrificed and blood samples were collected from the inferior vena cava, and lung tissues were harvested.Serum tumor necrosis factor-alpha (TNF-α) and syndecan-1 levels were measured by enzyme-linked immunosorbent assay, HMGB1 mRNA expression in lung tissues was detected using quantitative real-time polymerase chain reaction, wet/dry lung weight ratio was measured, and pulmonary vascular endothelial structure was observed with a transmission electron microscope. Results:Compared with group S, the serum TNF-α and syndecan-1 concentrations and wet/dry lung weight ratio were significantly increased, and the expression of HMGB1 mRNA in lung tissues was up-regulated in C and L groups ( P<0.05). Compared with group C, the serum TNF-α and syndecan-1 concentrations and wet/dry lung weight ratio were significantly decreased, and the expression of HMGB1 mRNA in lung tissues was down-regulated in group L ( P<0.05). The results of transmission electron microscope showed that the pulmonary vascular endothelium was intact and continuous, and the structure was dense in group S; the pulmonary vascular endothelium was discontinuous, and the structure was significantly loose in group C; the pulmonary vascular endothelium was basically intact, and the structure was slightly loose in group L. Conclusions:Lidocaine may reduce pulmonary vascular endothelial dysfunction in septic rats by inhibition of HMGB1 mRNA expression.

The expression of high mobility group protein 1 and interleukin-17 in peripheral blood and membrane tissues of pregnant women with premature rupture of membranes and its relationship with intrauterine infection / 中国医师进修杂志

Objective:To investigate the expression of high mobility group protein 1 (HMGB1) and interleukin-17 (IL-17) in peripheral blood and membrane tissues of pregnant women with premature rupture of membranes (PROM) and its relationship with intrauterine infection.Methods:Seventy-four pregnant women with PROM from January 2019 to June 2021 were selected as the study group, and 58 healthy pregnant women at the corresponding period were selected as the healthy control group. The levels of HMGB1 and IL-17 in peripheral blood and membrane tissues and serum CD 8+ were compared between the two groups. The pregnant women with PROM were divided into the chorioamnionitis group, subclinical chorioamnionitis group and normal group according to their intrauterine infection, the expression levels of HMGB1 and IL-17 in peripheral blood and membrane tissues of patients with different infection degrees were compared, and the correlation with the severity of intrauterine infection were analyzed. Results:The levels of peripheral blood HMGB1, membrane tissues HMGB1, peripheral blood IL-17, membrane tissues IL-17 and serum CD 8+ in the study group were higher than those in the control group: (28.34 ± 5.16) μg/L vs. (22.51 ± 4.09) μg/L, 0.79 ± 0.12 vs. 0.34 ± 0.05, (13.05 ± 2.57) ng/L vs. (8.16 ± 1.38) ng/L, 0.37 ± 0.06 vs. 0.12 ± 0.02, 0.386 ± 0.052 vs. 0.252 ± 0.044, there were statistical differences ( P<0.05). The levels of HMGB1 and IL-17 in peripheral blood and membrane tissues and serum CD 8+ were increased with the severity of severity of intrauterine infection ( P<0.05). The results of Spearman correlation analysis showed that the level of peripheral blood HMGB1, membrane tissues HMGB1 and IL-17 had positively correlated with the severity of intrauterine infection ( r = 0.336, 0.316, 0.311, P<0.05). The results of receiver operating characteristic curve analysis showed that combined detection of HMGB1 and IL-17 levels in peripheral blood and membrane tissues and serum CD 8+ levels in evaluating the severity of intrauterine infection had higher area under the curve than that of each index alone ( P<0.05). Conclusions:Pregnant women with PROM have abnormal HMGB1 and IL-17 levels in peripheral blood and membrane tissues, and HMGB1 levels in peripheral blood and mRNA expressions of HMGB1 and IL-17 in membrane tissues are positively correlated with the severity of intrauterine infection, which has evaluation value for the severity of the disease.

Research progress in mechanism of high mobility group box 1 regulating inflammatory response / 中华创伤杂志

Inflammatory reaction dominated by defense response will arise against infection and trauma. As an important proinflammatory cytokine, high mobility group box 1 (HMGB1) is widely expressed in all nuclear cells to mediate the inflammatory response. However, the biological functions of HMGB1 in inflammation vary depending on the type of HMGB1 protein modification and the localization in the cell. HMGB1 protein will be modified as acetylation of lysine residues, methylation of lysine residues, oxidation of cysteine residues, phosphorylation of serine residues, glycosylation of asparagine residues, adenosine diphosphate-ribosylation and lactylation of the protein in the nucleus, migrate from the nucleus to the cytoplasm, and release into the extracellular compartment. Extracellular HMGB1 can bind to receptors for advanced glycation end products (RAGE) and Toll-like receptors, activate cells and regulate inflammatory responses. The authors review the research progress in regulatory mechanism of HMGB1 in inflammation response from aspects of its post-translational modifications, releases, biological roles and binding receptors, hoping to provide theoretical basis for finding the targets of inflammation intervention.

Relationship between high mobility group protein box-1 level in peripheral blood and no-reflow in patients with acute myocardial infarction after interventional therapy / 中国基层医药

Objective:To investigate the relationship between high mobility group protein box-1 (HMGB1) level in peripheral blood and no-reflow in patients with acute myocardial infarction after interventional therapy.Methods:120 patients with acute myocardial infarction patients who received interventional treatment in Liuzhou People's Hospital, China between October 2019 and October 2020 were included in this study. They were divided into normal blood flow group ( n = 78) and no-reflow group ( n = 42) according to the situation of coronary reflow after interventional treatment. The clinical data and laboratory test results were compared between the two groups. The level of HMGB1 in peripheral blood was detected using enzyme-linked immunosorbent assay and compared between the two groups. The diagnostic value of HMGB1 in no reflow was analyzed by the receiver operating characteristic (ROC) curve. Results:There were no significant differences in age, gender, history of hypertension, history of smoking, history of coronary heart disease, and peak value of creatine kinase MB between no reflow and normal flow groups (all P > 0.05). The number of patients developing diabetes mellitus, the proportion of patients developing lesions of multiple vessels, C-reactive protein level and brain natriuretic peptide level in no-reflow group were significantly higher than those in the normal blood flow group (all P < 0.05). Enzyme-linked immunosorbent assay results revealed that there was significant difference in HMGB1 level between no reflow and normal flow groups [(5.2 ± 0.85) mg/L vs.(3.2 ± 0.9) mg/L, t = -2.38, P = 0.02). The ROC curve was used to compare the diagnostic values of HMGB1, brain natriuretic peptide and C-reactive protein for no reflow. The results showed that the area under the ROC curve values of HMGB1, brain natriuretic peptide and C-reactive protein were 0.746 (0.661-0.830), 0.605 (0.504-0.705) and 0.688 (0.595-0.781), respectively. The area under the ROC curve value of HMGB1 was the highest. Conclusion:The level of HMGB1 is obviously increased in patients with acute myocardial infarction presenting with no reflow, which has a high diagnostic value for no reflow.

Effects of anesthesia depth control on cognitive function and high mobility group protein B-1 level in older adult patients with breast cancer / 中国基层医药

Objective:To investigate the effects of anesthesia depth control on cognitive function and high mobility group protein B1 (HMGB-1) level in older adult patients with breast cancer.Methods:Eighty-six female older adult patients with breast cancer who received mastectomy between June 2019 and June 2020 in the First Hospital of China Medical University, China were included in this study. They were randomly assigned to undergo anesthesia with sodium phenobarbital and atropine at deep (bispectral index 30-45, deep anesthesia group, n = 43) or superficial level (bispectral index 45-60, superficial anesthesia group, n = 43). The mean arterial pressure, heart rate, HMGB-1 level, Mini-Mental State Examination (MMSE) score were assessed in each group. Results:There were no significant differences in mean arterial pressure and heart rate recorded during each time period between the deep anesthesia and superficial anesthesia groups (all P > 0.05). No significant difference in HMGB-1 level was found between the two groups before anesthesia induction and at the end of surgery (both P > 0.05). At 1 and 2 days after surgery, HMGB-1 level in the deep anesthesia group was (75.46 ± 3.33) pg/mL and (93.98 ± 4.32) pg/mL, respectively, which was significantly lower than that in the superficial anesthesia group [(87.89 ± 5.13) pg/mL and (121.01 ± 4.36) pg/mL, t = 13.327, 28.878, both P < 0.05)]. At 1 day before surgery, there was no significant difference in MMSE score between the two groups ( P > 0.05). In the deep anesthesia group, MMSE score was (26.73 ± 1.11) points, (28.16 ± 0.72) points, and (28.97 ± 0.88) points at 1, 3 and 6 days after surgery respectively, which was significantly higher than that in the superficial anesthesia group [(21.03 ± 1.46) points, (22.39 ± 1.24) points, and (24.69 ± 0.57) points, t = 20.380, 26.388, 26.768, all P < 0.05]. Conclusion:Deep anesthesia for mastectomy in older adult patients can reduce cognitive impairment and decrease HMGB-1 level after surgery, and plays a positive role in postoperative recovery.

Comparison of perioperative plasma HMGB1 concentrations in patients undergoing laparoscopic radical resection of cervical cancer using different anesthetic regimens / 中华麻醉学杂志

Objective:To compare the perioperative plasma high-mobility group box 1 protein (HMGB1) concentrations in the patients undergoing laparoscopic radical resection of cervical cancer using different anesthetic regimens.Methods:Sixty-eight American Society of Anesthesiologists physical status Ⅰor Ⅱ patients, aged 34-68 yr, with body mass index of 19-24 kg/m 2, undergoing elective laparoscopic radical resection of cervical cancer, were divided into 2 groups ( n=34 each) using a random number table method: general anesthesia group (G group) and general anesthesia combined with epidural anesthesia group (GE group). In group G, anesthesia was induced with midazolam, etomidate and cisatracurium and maintained with remifentanil, propofol and cisatracurium.In group GE, an epidural catheter was placed at L 1, 2 interspace before induction of anesthesia, general anesthesia was performed after the anesthesia level reached T 6, and the method was similar to those previously described in group G. Patient-controlled intravenous analgesia was used after operation to maintain visual analog scale score ≤ 3 points.Peripheral venous blood samples were collected at 10 min before anesthesia (T 0), at the end of operation, and at 1, 24 and 48 h after operation (T 1-4) for determination of plasma concentrations of HMGB1, interferon-gamma (IFN-γ) and interleukin-4 (IL-4) (by enzyme-linked immunosorbent assay) levels of T lymphocyte subsets CD3 + , CD4 + and CD8 + and CD4 + /CD8 + ratio (by flow cytometry). Results:Compared with group G, the plasma concentrations of IFN-γ and IL-4 were significantly decreased at T 2, 4, the plasma concentration of HMGB1 was decreased at T 2-4, and the levels of CD3 + at T 2-4, CD4 + at T 2 and CD8 + at T 2, 3 and CD4 + /CD8 + ratio were increased in group GE ( P<0.05). Conclusion:The plasma HMGB1 concentration is lower, which has less impact on immune function of the patients undergoing laparoscopic radical resection of cervical cancer with the combination of general anesthesia and epidural anesthesia than that with general anesthesia alone.

miRNA-34a reverses cisplatin resistance of osteosarcoma through targeted inhibition of high mobility group box 1 / 肿瘤研究与临床

Objective:To investigate the related mechanism of miRNA-34a (miR-34a) reverses cisplatin resistance of osteosarcoma through targeted inhibition of high mobility group box 1 (HMGB1).Methods:The MG-63 cisplatin-resistant cell line (MG-63/DDP) was constructed by using dose escalation and intermittent action, and then the successfully constructed MG-63/DDP cells were treated with different concentrations of cisplatin (0, 1, 5, 10, 20, 30 μg/ml), and CCK-8 method was used to detect cell survival rate. The MG-63/DDP cells were transfected respectively and then randomly divided into two groups: the miR-34a overexpression vector group and the miR-34a empty expression vector (miR-34a-NC) group. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of miR-34a. Transfected cells were treated with different concentrations of cisplatin (0, 1, 5, 10, 20, 30 μg/ml), and CCK-8 method was used to detect cell survival rate, flow cytometry was used to detect cell apoptosis. The dual luciferase reporter gene experiment was used to verify whether miR-34a regulated the expression of HMGB1, and Western blotting method was used to detect the HMGB1 protein expression level of the transfected cells. MG-63/DDP cells were transfected respectively and then randomly divided into two groups: HMGB1 gene silencing vector (si-HMGB1) group and its negative control vector (si-NC) group. Western blotting method was used to detect HMGB1 protein expression level and CCK-8 method was used to detect cell survival rate.Results:The MG-63/DDP cell line was successfully constructed. The survival rate of MG-63/DDP cells was higher than that of MG-63 cells when cells were treated with different concentrations of cisplatin (all P < 0.01), and half inhibitory concentration ( IC50) value of MG-63/DDP cells and MG-63 cells on cisplatin was 25.80 μg/ml and 0.47 μg/ml, respectively. qRT-PCR results showed that the relative expression level of miR-34a in MG-63/DDP cells was lower than that in MG-63 cells (0.46±0.04 vs. 1.02±0.05, t = 15.14, P < 0.01); compared with miR-34a-NC group, the relative expression of miR-34a in MG-63/DDP cells was increased in miR-34a overexpression vector group (1.67±0.09 vs. 1.00±0.02, t = -12.58, P < 0.01). Cell survival rate of miR-34a overexpression vector group and miR-34a-NC group was decreased with the increase in the concentration of cisplatin; cell survival rate of miR-34a overexpression vector group was lower than that of miR-34a-NC group when cells were treated with different concentrations of 5- 30 μg/ml cisplatin (all P < 0.01). The apoptotic rate of MG-63/DDP cells in miR-34a-NC group and miR-34a overexpression vector group was (25.1±1.7)% and (42.3±2.3)%, respectively when cells were treated with 20 μg/ml cisplatin; and in miR-34a overexpression vector group, MG-63/DDP cells had a higher rate of apoptosis ( P < 0.01). The dual luciferase reporter gene experiment results showed that compared with miR-34a-NC group, miR-34a overexpression vector group could inhibit the luciferase activity of PGL3- wild-type-HMGB1, and the difference was statistically significant ( t = 6.37, P < 0.01), while miR-34a overexpression vector group had no significant inhibitory effect on the luciferase activity of PGL3- mutant-HMGB1 ( t = 0.35, P = 0.74). The relative expression level of HMGB1 protein in miR-34a overexpression vector group was lower than that in miR-34a-NC group (0.43±0.02 vs. 1.00±0.14, t = 6.98, P < 0.01). Compared with si-NC group, the relative expression level and IC50 value of HMGB1 protein in si-HMGB1 group were reduced (all P < 0.05). Conclusion:Overexpression of miR-34a can enhance the chemosensitivity of osteosarcoma cells MG-63/DDP to cisplatin, and its mechanism may be related to the inhibition of HMGB1 expression.

HMGA Proteins in Stemness and Differentiation of Embryonic and Adult Stem Cells.

HMGA1 and HMGA2 are chromatin architectural proteins that do not have transcriptional activity per se, but are able to modify chromatin structure by interacting with the transcriptional machinery and thus negatively or positively regulate the transcription of several genes. They have been extensively studied in cancer where they are often found to be overexpressed but their functions under physiologic conditions have still not been completely addressed. Hmga1 and Hmga2 are expressed during the early stages of mouse development, whereas they are not detectable in most adult tissues. Hmga overexpression or knockout studies in mouse have pointed to a key function in the development of the embryo and of various tissues. HMGA proteins are expressed in embryonic stem cells and in some adult stem cells and numerous experimental data have indicated that they play a fundamental role in the maintenance of stemness and in the regulation of differentiation. In this review, we discuss available experimental data on HMGA1 and HMGA2 functions in governing embryonic and adult stem cell fate. Moreover, based on the available evidence, we will aim to outline how HMGA expression is regulated in different contexts and how these two proteins contribute to the regulation of gene expression and chromatin architecture in stem cells.

Disrupting the Homeostasis of High Mobility Group Protein Promotes the Systemic Movement of Bamboo mosaic virus.

Viruses hijack various organelles and machineries for their replication and movement. Ever more lines of evidence indicate that specific nuclear factors are involved in systemic trafficking of several viruses. However, how such factors regulate viral systemic movement remains unclear. Here, we identify a novel role for Nicotiana benthamiana high mobility group nucleoprotein (NbHMG1/2a) in virus movement. Although infection of N. benthamiana with Bamboo mosaic virus (BaMV) decreased NbHMG1/2a expression levels, nuclear-localized NbHMG1/2a protein was shuttled out of the nucleus into cytoplasm upon BaMV infection. NbHMG1/2a knockdown or even overexpression did not affect BaMV accumulation in inoculated leaves, but it did enhance systemic movement of the virus. Interestingly, the positive regulator Rap-GTPase activation protein 1 was highly upregulated upon infection with BaMV, whereas the negative regulator thioredoxin h protein was greatly reduced, no matter if NbHMG1a/2a was silenced or overexpressed. Our findings indicate that NbHMG1/2a may have a role in plant defense responses. Once its homeostasis is disrupted, expression of relevant host factors may be perturbed that, in turn, facilitates BaMV systemic movement.

Effects of oxymatrine on the expression and release of high mobility group box 1 in pancreatic acinar cells stimulated by hydrogen peroxide / 中华胰腺病杂志

Objective@#To observe the effects of oxymatrine(OM) on the expression and release of high mobility group box 1(HMGB1) in rat pancreatic acinar cell line AR42J stimulated by hydrogen peroxide.@*Methods@#MTT method was used to detect the effects of H2O2 in different concentrations on the survival of AR42J cells. AR42J cells cultured in vitro were divided into control group, H2O2 group and H2O2+ OM group. An equal volume of H2O2(final concentration 0.16 mmol/L) was added in H2O2 group and H2O2+ OM group, respectively, while an equal volume of triple distilled water was added in control group. In H2O2+ OM group, OM(final concentration 0.5 g/L)was added 0.5 h before the addition of H2O2, and cell samples and supernatant were collected after 24 h culture. The expression of HMGB1 protein was detected by Western blotting, the level of HMGB1 protein in cell supernatant was detected by enzyme-linked immunosorbent assay, and the intracellular distribution of HMGB1 protein was detected by immunofluorescence.@*Results@#In the H2O2 group, the expression of HMGB1, the secretion of HMGB1 in the supernatant and the proportion of cytoplasmic HMGB1 in the total HMGB1 were significantly higher than those in the control group[1.04±0.04 vs 0.69±0.02, (4.84±0.13)μg/L vs (2.68±0.07)μg/L, (35.7±2.5)% vs (10.7±1.9)%], and all the differences were statistically significant (all P<0.05). After OM intervention, HMGB1 protein expression, secretion and cytoplasmic proportion were [0.82±0.02, (3.97±0.10)μg/L and (27.3±1.7)%], respectively, which were obviously lower than those in the H2O2 group, and all the differences were statistically significant (all P<0.05).@*Conclusions@#H2O2 can induce the expression and release of HMGB1 in rat pancreatic acinar cells; OM treatment could alleviate the severity of oxidative stress injuries induced by H2O2 in AR42J cells.

[Relationship between expression of high-mobility group box-1 and inflammatory cytokines in patients with bone cancer pain].

Objective: To investigate the change and relationship between serum high-mobility group box-1(HMGB1) and related inflammatory cytokines level in patients suffer with bone metastatic pain. Methods: Collection of the bone cancer pain patients who received analgesic therapy the department of pain in The First Affiliated Hospital of Jiaxing University from November 2016 to August 2016. Serum concentration of HMGB1, the Receptor of Advanced Glycation Endproducts (RAGE), monocyte chemotactic protein-1(MCP-1), tumor necrosis factor -α (TNF-α), interleukin-1ß (IL-1ß), interleukin-10 (IL-10), interleukin-13 (IL-13), and transforming growth factor-ß (TGF-ß) levels were determined in 15 healthy individuals as healthy donor and 15 patients with bone metastatic pain by enzyme-linked immunosorbent (ELISA) . The healthy individuals and patients with bone metastatic pain were collected before treatment and on 7 d after the treatment. Results: The serum concentration of HMGB1 and RAGE were significantly increased in tumorous group compared with healthy group[(8.8±2.3) vs (1.9±1.1) µg/L,(231±16) vs (46±20) ng/L); t=7.10,12.44, both P<0.05], then decreased after analgesic therapy [(4.77±1.36) µg/L, (129.80±29.32) ng/L, t=7.10, 12.44, both P<0.05]. The serum concentration of proinflammatory cytokines such as MCP-1, TNF-α, and IL-1ß were significantly increased in tumorous group when compared with healthy group, and decreased after analgesic therapy (all P<0.05). The expression of anti-inflammatory cytokines such as IL-10, IL-13, and TGF-ß were significantly increased in tumorous group when compared with healthy group, and decreased after analgesic therapy (all P<0.05).Compared with healthy group, the levels of MCP-1/IL-10, MCP-1/IL-13, MCP-1/TGF-ß, TNF-α/IL-10, TNF-α/IL-13, TNF-α/TGF-ß, IL-1ß/IL-10, IL-1ß/IL-13, IL-1ß/TGF-ß were significantly increased in tumorous group (all P<0.05). Conclusion: HMGB1 may adjust the proinflammatory-anti-inflammatory system homeostasis to participate in the development of bone metastatic pain.

Clinical significance of serum high mobility group box-1 protein in patients with acute cholangitis / 中华普通外科杂志

Objective@#To analyze perioperative serum high mobility group box-1 protein (HMGB1) levels in patients with acute cholangitis and its clinical significance.@*Methods@#118 cases of choledocholithiasis with acute cholangitis were retrospectively analyzed, admittd in Minhang Hospital from Jan 2017 to Dec 2017. Enzyme linked immunosorbent assay (ELISA) was used to detect serum HMGB1 levels before and after ERCP. The relationship between serum HMGB1 levels and severity of the disease was analyzed.@*Results@#The serum HMGB1 levels in the healthy controls, mild cholangitis group, moderate cholangitis group and severe cholangitis group were(1.74±0.79) μg/L, (9.19±4.86) μg/L, (12.62±4.13) μg/L, (18.02±3.84) μg/L, respectively. The serum HMGB1 levels were significantly different in these four groups (F=63.348, P<0.05). The serum HMGB1 levels in the three groups after ERCP were significantly lower than those before ERCP(t=10.978, t=35.682, t=42.649; P<0.05). The serum HMGB1 levels were positively correlated with WBC, CRP, total bilirubin, direct bilirubin and alanine aminotransferase.@*Conclusion@#Serum HMGB1 levels were significantly higher in patients with acute cholangitis, and the more serious the disease, the higher the HMGB1 levels. After ERCP, serum HMGB1 levels were significantly lower than before. HMGB1 is an effective parameter for evaluating the severity of acute cholangitis.

Clinical significance of serum high mobility group box-1 protein in patients with acute cholangitis / 中华普通外科杂志

Objective To analyze perioperative serum high mobility group box-1 protein (HMGB1) levels in patients with acute cholangitis and its clinical significance.Methods 118 cases of choledocholithiasis with acute cholangitis were retrospectively analyzed,admittd in Minhang Hospital from Jan 2017 to Dec 2017.Enzyme linked immunosorbent assay (ELISA) was used to detect serum HMGB1 levels before and after ERCP.The relationship between serum HMGB1 levels and severity of the disease was analyzed.Results The serum HMGB1 levels in the healthy controls,mild cholangitis group,moderate cholangitis group and severe cholangitis group were(1.74 ±0.79) μg/L,(9.19 ±4.86) μg/L,(12.62 ± 4.13) μg/L,(18.02 ±3.84) μg/L,respectively.The serum HMGB1 levels were significantly different in these four groups (F =63.348,P ＜ 0.05).The serum HMGB1 levels in the three groups after ERCP were significantly lower than those before ERCP (t =10.978,t =35.682,t =42.649;P ＜ 0.05).The serum HMGB1 levels were positively correlated with WBC,CRP,total bilirubin,direct bilirubin and alanine aminotransferase.Conclusion Serum HMGB1 levels were significantly higher in patients with acute cholangitis,and the more serious the disease,the higher the HMGB1 levels.After ERCP,serum HMGB1 levels were significantly lower than before.HMGB1 is an effective parameter for evaluating the severity of acute cholangitis.

Effect of propofol on HMGB1/TLR4 signaling pathway during hepatic ischemia-reperfusion injury in rats / 中华麻醉学杂志

Objective To evaluate the effect of propofol on high-mobility group box 1 protein (HMGB1)/Toll-like receptor 4 (TLR4) signaling pathway during hepatic ischemia-reperfusion (I/R) injury in rats.Methods Thirty-six clean-grade healthy male Sprague-Dawley rats,aged 3 months,weighing 250 -300 g,were divided into 3 groups (n=12 each) using a random number table method:sham operation group (group S),hepatic I/R group (group I/R) and propofol group (group P).Hepatic I/R injury was induced by occluding the portal vein and hepatic artery supplying the left and middle lobes of the liver for 1 h followed by 6-h reperfusion in anesthetized rats.Propofol was infused via the tail vein at a rate of 12 mg ·kg-1 · h-1 starting from 20 min before ischemia until 6 h of reperfusion in group P.The rats were sacrificed at 6 h of reperfusion,and the left lobe of the liver was removed for microscopic examination of the pathological changes which were scored and for determination of the expression of HMGB1,TLR4,tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-6) in liver tissues (by Western blot).Results Compared with group S,pathological scores of liver tissues were significantly increased,and the expression of HMGB1,TLR4,TNF-α and IL-6 was up-regulated in I/R and P groups (P＜0.05).Compared with group I/R,pathological scores of liver tissues were significantly decreased,and the expression of HMGB1,TLR4,TNF-α and IL-6 was down-regulated in group P (P＜ 0.05).Conclusion The mechanism by which propofol reduces liver I/R injury is associated with blocking HMGB-1/TLR4 signaling pathway and inhibiting inflammatory responses in rats.

Biological characteristics of high-mobility group box-1 and its role in the development, progression, and treatment of hepatitis B virus-related liver diseases / 临床肝胆病杂志

High-mobility group box-1 (HMGB1) is a member of the high-mobility group proteins and is present in eukaryotic cells. HMGB1 is not only a nuclear protein but also a pro-inflammatory factor, and the increase in HMGB1 level in the body indicates cell destruction and inflammatory response. Hepatitis B virus (HBV) infection is a major cause of chronic hepatitis B, liver cirrhosis, and hepatocellular carcinoma in the world. In recent years, the role of HMGB1 in HBV-related liver diseases has attracted more and more attention, especially the important role of HMGB1 in the progression of liver inflammation and liver cancer. This article reviews the recent research advances in the role of HMGB1 in the development, progression, and treatment of HBV-related liver diseases.

Noninvasive biomarkers of premature delivery: Research progress and application prospect / 第二军医大学学报

Preterm birth is one of the most common obstetric complications. In recent decades, the incidence of premature birth remains high throughout the world, and even shows a rising trend in some countries and areas. Therefore, understanding the mechanisms of human parturition and developing the effective prevention and treatment strategies of premature delivery are urgent to improve maternal and fetal health and overall quality of population. Due to the limitation of ethics and testing methods, the prediction and early diagnosis of premature birth have been the primary problem that perplexes obstetricians. Various body fluids, including amniotic fluid, cervicovaginal fluid, urine, saliva and blood, provide rich sources of putative biomarkers that may be causative or reflective of preterm labor. In recent years, the exploration of novel biomarkers for noninvasive detection is in the ascendant, which sheds new lights for the prediction and early diagnosis of preterm labor. This review compares the biomarkers for the detection of preterm birth, and discusses the future prospect.