[Interpretation of Chinese expert consensus on the surgical treatment for adenocarcinoma of esophagogastric junction(2024 edition)].

Due to the unique nature of its anatomical location, the adenocarcinoma of esophagogastric junction (AEG) has been a subject of controversy and disagreement including its definition, staging, and treatment strategies. Chinse expert Consensus on Surgical Treatment of Adenocarcinoma of Esophagogastric Junction in China (2018 Edition) had been released in September 2018 and had played a pioneering role in unifying thoracic and general surgeons in China on surgical treatment strategies for AEG. Over the past five years, the emergence of several clinical research results on AEG has provided new clinical evidence for the selection of key surgical treatment strategies. Therefore, to further standardize the surgical treatment of AEG in China, Chinese Expert Consensus on Surgical Treatment of Adenocarcinoma of Esophagogastric Junction in China (2024 Edition) was released in 2024 by Chinese expert panel including 25 gastrointestinal surgeons and 24 thoracic surgeons. Based on the highest-level clinical research evidence in recent 5 years, this consensus ultimately formulates 29 recommendations on hotspots and key points on surgical treatment of AEG and summary 5 issues that are still awaiting further exploration. This review will provide a summary and detailed interpretation of the recommendations outlined in this consensus.

[Expression of endosialin in human hypertrophic scars and its regulation on fibroblast phenotype].

Objective: To explore the expression of endosialin, i.e., CD248 in human hypertrophic scars (HSs) and its regulatory effect on the phenotype of hypertrophic scar fibroblasts (HSFs). Methods: The method of experimental research was used. From March to May, 2023, 3 pediatric patients with HS were admitted to the Department of Burns and Cutaneous Surgery of the First Affiliated Hospital of Air Force Medical University, including 2 females and 1 male, aged one year ten months to two years. The HS tissue resected during the surgery and the remaining full-thickness skin graft, i.e., normal skin tissue after full-thickness skin grafting were collected from the aforementioned pediatric patients for subsequent experiments. Using the aforementioned two types of tissue, the histological structures were observed by hematoxylin-eosin staining, collagen distribution was observed by Masson staining, and the expression of CD248 was observed and measured by immunohistochemical staining. The primary HSFs were isolated from HS tissue using explant culture technique, and the 3rd to 5th passages of HSFs were used in subsequent experiments. According to the random number table, HSFs were divided into immunoglobulin G78 (IgG78)-treated group and IgG control group, which were treated with 200 nmol/L human CD248 monoclonal antibody IgG78 and human IgG control antibody for 24 h, respectively. The mRNA expressions of collagen type Ⅰ (Col Ⅰ) and α-smooth muscle actin (α-SMA) in HSFs were measured by real-time fluorescence quantitative reverse transcription polymerase chain reaction, the protein expressions of Col Ⅰ and α-SMA in HSFs were detected by Western blotting, and the intracellular location and protein expressions of Col Ⅰ and α-SMA were detected by immunofluorescence method. The number of samples in each experiment was 3. Data were statistically analyzed with paired sample t test and independent sample t test. Results: Compared with those in normal skin tissue, the epidermis and dermis in HS tissue were significantly thicker, with massive accumulation and disordered arrangement of collagen in the dermis. The expression of CD248 in HS tissue was significantly upregulated compared with that in normal skin tissue (t=5.29, P<0.05). At post treatment hour 24, the mRNA expressions of Col Ⅰ and α-SMA of HSFs in IgG78-treated group were 0.39±0.05 and 0.56±0.09, respectively, which were significantly lower than 1.00±0.07 and 1.00±0.08 in IgG control group, respectively (with t values of 11.87 and 6.49, respectively, P values all <0.05). The protein expressions of Col Ⅰ and α-SMA of HSFs in IgG78-treated group were 0.617±0.011 and 0.67±0.14, respectively, which were significantly lower than 1.259±0.052 and 1.23±0.16 in IgG control group, respectively (with t values of 20.92 and 4.52, respectively, P values all <0.05). At post treatment hour 24, immunofluorescence staining showed that Col Ⅰ and α-SMA mainly located in the cytoplasm of HSFs in the two groups, and the protein expressions of Col Ⅰ and α-SMA of HSFs in IgG78-treated group were obviously downregulated compared with those in IgG control group. Conclusions: The expression of CD248 is significantly upregulated in human HS. Targeted blockade of CD248 can significantly inhibit the collagen synthesis by HSFs and the transdifferentiation of HSFs into myofibroblasts.

[Identification the key factor of pulmonary fibrosis following silica nanoparticles exposure based on bioinformatics analysis].

Objective: To investigate the main mechanisms of pulmonary fibrosis following silica nanoparticles (SiNPs) exposure through constructing the macrophage-fibroblast model in vitro, which simulated the process of pulmonary fibrosis. Methods: In January 2021, human mononuclear leukemia cells (THP-1) were treated with 0, 25, 50, 100 µg/ml SiNPs for 24 h. The supernatant of THP-1 cells was collected and applied to human embryonic lung fibroblast cells (MRC-5) which divided into control and low, medium and high dose groups at the logarithmic growth stage for 24 h. MRC-5 cell viability was detected by CCK8. The hydroxyproline (Hyp), interleukin 6 (IL-6), interleukin 1 beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α) expression were detected in the supernatants of MRC-5. The changed proteins were detected by liquid-phase mass spectrometry in high dose group. GeneCard database were applied to identity the differential pulmonary fibrosis proteins in high dose group. Gene Ontology (GO) was performed to identity the key biological process in differential pulmonary fibrosis proteins of high dose group. The String database was used to construct the protein-protein interactions (PPI) network of differential pulmonary fibrosis proteins. The APP of CytoHubba was applied to calculate the key protein of differential pulmonary fibrosis proteins in PPI network. Correlation coefficients between key differential pulmonary fibrosis proteins were calculated using Pearson correlation analysis. Western blotting was applied to detect the expression of key proteins of differential pulmonary fibrosis proteins in different groups. Results: CCK8 results showed that MRC-5 cell viability was increasing in low, medium and high dose groups compared with control group (P<0.05). The expression levels of Hyp and IL-1ß in different group were increased compared with control group, the expression levels of IL-6 and TNF-α were increased in high dose group compared with control group (P<0.05). GeneCard database identified 26 differential pulmonary fibrosis proteins, which were mainly involved in extracellular matrix hydrolysis, cell inflammatory response, tissue repair, cell proliferation, inflammation response by GO analysis. The APP of CytoHubba was calculated that matrix metalloproteinase 9 (MMP9) and tissue inhibitor metalloproteinase 1 (TIMP1) played an important role in PPI network. The results of correlation analysis showed that MMP9 was correlated with the expression of matrix metalloproteinase 1 (MMP1), matrix metalloproteinase 3 (MMP3), TIMP1 and epidermal growth factor receptor (EGFR) (r=0.97, 0.98, 0.94, 0.93, P<0.05). Western blotting results showed that TIMP1 protein expression was increased in low, medium and high dose groups, while MMP9 protein expression was increased only in high dose group (P<0.05) . Conclusion: Differential expression proteins related with pulmonary fibrosis in MRC-5 cells mainly regulate biological processes of extracellular matrix hydrolysis, tissue repair, and cellular inflammation response following SiNPs exposure. MMP9 and TIMP1 may be the key proteins, which affected the fibrosis process in vitro pulmonary fibrosis model.

[Intravenous infusion of methylene blue to visualize the ureter in laparoscopic colorectal surgery].

Objective: Intraoperative localization of the ureter can contribute to accurate dissection and minimize ureteral injury in colorectal surgery. We aim to summarize a single center's experience of fluorescence ureteral visualization using methylene blue (MB) and explore its visualization efficiency. Methods: This is a descriptive case-series-study. Clinical data of patients who had undergone laparoscopic colorectal surgery and fluorescence visualization of the ureter in the Gastrointestinal Surgery Department of Guangdong Provincial People's Hospital from March 2022 to May 2022 were retrospectively collected. Patients with incomplete surgery videos, renal insufficiency, or allergic reactions were excluded. MB was infused with 0.9% NaCl at 1.0 mg/kg in 100 mL of normal saline for 5 to 15 minutes during laparoscopic exploration. Imaging was performed using a device developed in-house by OptoMedic (Guangdong, China) that operates at 660nm to achieve excitation of MB. Clinical information, MB dosage, rate of successful fluorescence, time to fluorescence, operation time, blood loss, intraoperative blood oxygen levels, pathological staging, changes in renal function, and post-operative complications were retrospectively analyzed. Results: The study cohort comprised 27 patients (24 men and 3 women) with an average age of (60.25±16.95) years and an average body mass index of (21.72±3.42) kg/m2. The dosage of MB was 0.3-1.0 mg/kg and the infusion time was 5-15 minutes. Fluorescence signals were detected in all patients. The median time to signal detection was 20 (range, 10 to 40) minutes after MB infusion. The range of intraoperative blood oxygen fluctuation averaged 2.5% (range, 0 to 7.0%). The median change in creatine concentration was -1.3 (range, -17.2 to 29.2) µmol/L. No patients had complications associated with use of MB. Fluorescence visualization of the ureter was very valuable clinically in two patients (thick mesentery, stage T4). Conclusion: MB is a safe and effective means of visualizing the ureter by fluorescence during laparoscopic colorectal surgery, especially when the procedure is difficult. MB in a dosage of less than 1 mg/kg can slowly infused for more than 5 minutes during laparoscopic exploration. During the infusion, attention must be paid to blood oxygen fluctuations.

Quantitative 3D Characterization for Kinetics of Corrosion Initiation and Propagation in Additively Manufactured Austenitic Stainless Steel.

In situ X-ray computed tomography (X-ray CT) is used to investigate the effects of characteristic microstructural features on the pitting initiation and propagation in austenitic stainless steel specimens prepared with laser powder bed fusion (LPBF) additive manufacturing. In situ X-ray CT in probing the mechanism and kinetics of localized corrosion is demonstrated by immersing two LPBF specimens with different porosities in an aggressive ferric chloride solution for the evaluation of corrosion. X-ray CT images are acquired from the specimens after every 8 hours of immersion over an extended period of time (216 hours). Corrosion pit growth is then quantitatively analyzed with a data-constrained modeling method. The pitting growth mechanism of LPBF stainless steel is found to be different from that of conventional stainless steels. More specifically, the mechanism of corrosion pit initiation is closely correlated with the original lack of fusion porosity (LOF) distribution on the surface of the specimens and preferential pit propagation through the LOF pores inside the specimens. Pit growth kinetics are derived from pit volume changes determined through 3D data analysis. The pit growth kinetics in LPBF specimens are found to vary in the initial pit formation, competitive pit propagation, and the dominant pit growth stages.

[Effects of mechanical tension on the formation of hypertrophic scars in rabbit ears and transforming growth factor-ß1/Smad signaling pathway].

Objective: To explore the effects of mechanical tension on the formation of hypertrophic scars in rabbit ears and transforming growth factor-ß1 (TGF-ß1)/Smad signaling pathway. Methods: The experimental research method was adopted. Six New Zealand white rabbits, male or female, aged 3-5 months were used and 5 full-thickness skin defect wounds were made on the ventral surface of each rabbit ear. The appearance of all rabbit ear wounds was observed on post surgery day (PSD) 0 (immediately), 7, 14, 21, and 28. On PSD 28, the scar formation rate was calculated. Three mature scars in the left ear of each rabbit were included in tension group and the arch was continuously expanded with a spiral expander. Three mature scars in the right ear of each rabbit were included in sham tension group and only the spiral expander was sutured without expansion. There were 18 scars in each group. After mechanical tension treatment (hereinafter referred to as treatment) for 40 days, the color and texture of scar tissue in the two groups were observed. On treatment day 40, the scar elevation index (SEI) was observed and calculated; the histology was observed after hematoxylin eosin staining, and the collagen morphology was observed after Masson staining; mRNA expressions of TGF-ß1, Smad3, collagen Ⅰ, collagen Ⅲ, and α-smooth muscle actin (α-SMA) in scar tissue were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction; and the protein expressions of TGF-ß1, collagen Ⅰ, collagen Ⅲ, and α-SMA, and phosphorylation level of Smad3 in scar tissue were detected by Western blotting. The number of samples of each group in the experiments was 3. Data were statistically analyzed with independent sample t test. Results: On PSD 0, 5 fresh wounds were formed on all the rabbit ears; on PSD 7, the wounds were scabbed; on PSD 14, most of the wounds were epithelialized; on PSD 21, all the wounds were epithelialized; on PSD 28, obvious hypertrophic scars were formed. The scar formation rate was 75% (45/60) on PSD 28. On treatment day 40, the scar tissue of rabbit ears in tension group was more prominent than that in sham tension group, the scar tissue was harder and the color was more ruddy; the SEI of the scar tissue of rabbit ears in tension group (2.02±0.08) was significantly higher than 1.70±0.08 in sham tension group (t=5.07, P<0.01). On treatment day 40, compared with those in sham tension group, the stratum corneum of scar tissue became thicker, and a large number of new capillaries, inflammatory cells, and fibroblasts were observed in the dermis, and collagen was more disordered, with nodular or swirling distribution in the scar tissue of rabbit ears in tension group. On treatment day 40, the mRNA expressions of TGF-ß1, Smad3, collagen Ⅰ, collagen Ⅲ, and α-SMA in the scar tissue of rabbit ears in tension group were respectively 1.81±0.25, 5.71±0.82, 7.86±0.56, 4.35±0.28, and 5.89±0.47, which were significantly higher than 1.00±0.08, 1.00±0.12, 1.00±0.13, 1.00±0.14, and 1.00±0.14 in sham tension group (with t values of 5.36, 9.82, 20.60, 18.26, and 17.13, respectively, all P<0.01); the protein expressions of TGF-ß1, collagen Ⅰ, collagen Ⅲ, and α-SMA, and phosphorylation level of Smad3 in the scar tissue of rabbit ears in tension group were respectively 0.865±0.050, 0.895±0.042, 0.972±0.027, 1.012±0.057, and 0.968±0.087, which were significantly higher than 0.657±0.050, 0.271±0.029, 0.631±0.027, 0.418±0.023, and 0.511±0.035 in sham tension group (with t values of 5.08, 21.27, 15.55, 16.70, and 8.40, respectively, all P<0.01). Conclusions: Mechanical tension can inhibit the regression of hypertrophic scars in rabbit ears through stimulating the hyperplasia of scars, inhibiting the normal arrangement of dermal collagen fibers, and intensifying the deposition of collagen fibers, and the mechanism may be related to the activation of TGF-ß1/Smad signaling pathway by mechanical tension.

[Research progress of mast cell activation syndrome].

Mast cells are the main effector cells in allergic diseases. Allergic diseases are mostly a direct result of mast cell mediator release effects, while allergen activation is only one of many triggers for mast cell mediator secretion. Increased mast cell number, high mast cell reactivity, or both can lead to abnormal mast cell activation. Mast cell activated syndrome (MCAS) refers to a group or a"spectrum"of mediator-related, symptomatically similar diseases in which mast cells are stimulated by multiple factors. The symptoms and signs of mast cell disease overlap with allergic diseases, but the etiology is different, which requires clinical attention. This article summarizes the research progress on mast cell activation syndrome in recent years thus increase awareness of the differential diagnosis.

Oridonin Enhances the Anti-Metastasis Effect of Oxaliplatinliplatin on Colorectal Cancer Liver Metastasis.

The anti-metastasis effect of oridonin in combination with oxaliplatin on colorectal cancer liver metastasis was studied using a BALB/c nude mouse model. The liver condition, bloody ascites, cholestasis, and liver metastasis scores in the three groups receiving oxaliplatin combined with oridonin were significantly milder than in the control group and importantly the anti-migratory effect of oxaliplatin combined with oridonin was obviously the strongest (p<0.05). Oridonin possessed no hepatotoxicity; instead, it effectively alleviated liver injury caused by oxaliplatin. Oridonin alone or in combination with oxaliplatin significantly decreased serum levels of α-fetoprotein and carcinoembryonic antigen. Therefore, oridonin combined with oxaliplatin displays great potential to markedly increase the anti-metastasis effect of oxaliplatin in the treatment of liver metastases of colorectal cancer.

[Patterns of recurrence after neoadjuvant chemoradiotherapy compared with surgery alone in esophageal cancer].

Associated with improvement in survival, the neoadjuvant therapy had become the mainstay of therapy for patients with locally advanced esophageal cancer. Despite a significantly better survival, the recurrence risk after neoadjuvant therapy remains considerably high, with recurrence rate of>40%. Thus, it's important to gain a thorough understanding of the recurrence patterns for developing effective tertiary prevention and follow-up strategies. The aim of this review was to compare the patterns of recurrence in patients with esophageal cancer who received preoperative therapy followed by surgery or surgery alone. It is found that the most frequent recurrence pattern was distant metastasis in esophageal cancer regardless receipt of neoadjuvant therapy or not, and the major effect of neoadjuvant therapy appears to be an improvement in local regional disease control without a reduction in systemic. This frustrating fact may explain the poor survival of esophageal cancer patients receiving neoadjuvant therapy.

[Correlation between preoperative inflammatory biomarkers and postoperative pneumonia or long-term prognosis in patients with esophageal cancer after neoadjuvant therapy].

Objective: To examine the correlation between neutrophil-lymphocyte ratio (NLR), lymphocyte-monocyte ratio (LMR) and neutrophil-monocyte ratio (NMR) for postoperative pneumonia or long-term overall survival in patients with esophageal cancer after neoadjuvant therapy. Methods: The clinical data of 137 patients, including 111 males and 26 females, with the age of (M(QR))61(10) years (range: 45 to 75 years), undergoing radical resection of esophageal cancer after neoadjuvant therapy admitted at Department of Thoracic Surgery, West China Hospital from January 2016 to May 2019 were analyzed retrospectively. The blood routine one or two days before surgery and the occurrence of pneumonia after surgery were collected via hospital information system. The absolute count of neutrophils, lymphocytes and monocytes was recorded, to calculate NLR, LMR and NMR. The survival of patients was recorded systematically via follow-up. In the first part, the influencing factors of postoperative inflammation were analyzed, to group the patients into two groups according to the occurrence of postoperative pneumonia. χ2 test, t-test or rank-sum test were conducted for inter-group comparison. In the second part, cutoff values of inflammatory biomarkers were obtained with the receiver operating characteristic (ROC) curve and grouped, with postoperative pneumonia as endpoint criteria. Independent factors correlated with postoperative pneumonia were determined through univariate and multivariate Logistic regression analysis. In the third part, the analysis on prognosis factors was carried on, with the survival as endpoint criteria. Cutoff values of inflammatory biomarkers were obtained with X-Tile software and grouped. The survival analysis was carried on with univariate and multivariate Cox proportional hazards regression model, and the Kaplan-Meier curve was drawn finally. The results of survival analysis were verified by Log-rank test. Results: Median follow-up time was 614 (299) days (range: 382 to 1 612 days). Cutoff values of NLR, LMR, and NMR obtained via the ROC curve were 3.0, 3.9, and 6.2, respectively. According to the multivariate Logistic regression analysis, NLR>3.0 (OR=2.740, 95% CI: 1.221 to 6.152, P=0.015) and LMR>3.9 (OR=0.140, 95% CI: 0.022 to 0.890, P=0.037) were independent prognosis factors for postoperative pneumonia in patients with esophageal cancer after neoadjuvant therapy. Cutoff values of NLR, LMR, and NMR obtained with X-Tile software were 3.3, 4.2, and 7.2, respectively. Through multivariate Cox proportional risk regression analysis, late tumor ypTNM staging (8th AJCC) (HR=2.087, 95% CI:1.079 to 4.038, P=0.029), poor pathologic response (HR=2.251, 95% CI: 1.117 to 4.538, P=0.023), and LMR>4.2 (HR=0.347, 95% CI: 0.127 to 0.946, P=0.039) could be independent prognosis factors for overall survival. Kaplan-Meier survival analysis indicated that the overall survival of patients with LMR ≤4.2 was worse (P=0.002), with the 1-year overall survival rate of 82.9%, and the 1-year overall survival rate of patients with LMR>4.2 was 94.6%. Conclusion: Preoperative LMR ≤3.9 and NLR>3.0 can be considered as independent prognosis factors for postoperative pneumonia, while LMR≤4.2 as one of independent prognosis factors for overall survival.

Transmembrane Protease TMPRSS4 Promotes the Formation and Development of Mismatch Repair Deficient Colon Cancer Liver Metastasis.

We studied metastasis-promoting effect of transmembrane protease TMPRSS4 on mismatch repair (MMR)-deficient colorectal cancer liver metastasis in BALB/c nude mouse model. Histomorphological and histopathological studies showed that the number of liver metastases in the study group were significantly higher than that in the control group (p<0.05). The expression of TMPRSS4 mRNA and protein in the study group were obviously higher than in the control group (p<0.05). These findings suggest that TMPRSS4 possesses a metastasis-promoting effect and its low expression can effectively block the progression of MMR-deficient colon cancer liver metastasis.

No pulsed radio emission during a bursting phase of a Galactic magnetar.

Fast radio bursts (FRBs) are millisecond-duration radio transients of unknown physical origin observed at extragalactic distances1-3. It has long been speculated that magnetars are the engine powering repeating bursts from FRB sources4-13, but no convincing evidence has been collected so far14. Recently, the Galactic magnetar SRG 1935+2154 entered an active phase by emitting intense soft γ-ray bursts15. One FRB-like event with two peaks (FRB 200428) and a luminosity slightly lower than the faintest extragalactic FRBs was detected from the source, in association with a soft γ-ray/hard-X-ray flare18-21. Here we report an eight-hour targeted radio observational campaign comprising four sessions and assisted by multi-wavelength (optical and hard-X-ray) data. During the third session, 29 soft-γ-ray repeater (SGR) bursts were detected in γ-ray energies. Throughout the observing period, we detected no single dispersed pulsed emission coincident with the arrivals of SGR bursts, but unfortunately we were not observing when the FRB was detected. The non-detection places a fluence upper limit that is eight orders of magnitude lower than the fluence of FRB 200428. Our results suggest that FRB-SGR burst associations are rare. FRBs may be highly relativistic and geometrically beamed, or FRB-like events associated with SGR bursts may have narrow spectra and characteristic frequencies outside the observed band. It is also possible that the physical conditions required to achieve coherent radiation in SGR bursts are difficult to satisfy, and that only under extreme conditions could an FRB be associated with an SGR burst.

[The progress in management of esophagogastric variceal bleeding in cirrhotic portal hypertension].

Esophagogastric variceal bleeding (EVB) is the most dangerous complication of cirrhotic portal hypertension.With the continuous emergence of research findings on EVB, multiple disciplinary team, including internal medicine department, surgery department, intervention therapy department, radiology department, has become a new mode for the prevention and treatment of EVB. This article first reviewed the classification of esophageal varices and gastric varices, and then reviewed the recent research findings of EVB from three aspects: primary prophylaxis, active variceal bleeding treatment, and secondary prophylaxis.The aim was to provide new ideas for the individualized prevention and treatment of EVB.

[Effects and mechanism of pyrroloquinoline quinine on mitochondrial function and cell survival of rat bone marrow mesenchymal stem cells under oxidative stress].

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(2)O(2)) alone group, and H(2)O(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(2)O(2) alone group were cultured in normal medium containing 200 µmol/L H(2)O(2) for 24 hours; the cells in H(2)O(2)+ PQQ group were pre-incubated with normal medium containing 100 µmol/L PQQ for 2 hours, and then with H(2)O(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(2)O(2) alone group, and H(2)O(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(2)O(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(2)O(2) alone group, the cell morphology of H(2)O(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(2)O(2) alone group was significantly increased to (37.1±2.0)% (t=10.57, P<0.01). Compared with that in H(2)O(2) alone group, the apoptosis rate of cells in H(2)O(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(2)O(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(2)O(2) alone group, the mitochondrial membrane potential of cells in H(2)O(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(2)O(2) alone group was disordered, with disappeared mitochondrial cristae and decreased mitochondrial matrix density. Compared with that in H(2)O(2) alone group, the mitochondrial structure of cells in H(2)O(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(2)O(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(2)O(2) alone group, the CAT activity of cells in H(2)O(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(2)O(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(2)O(2) alone group, the protein expression of Epac1 and the AMPK phosphorylation level of cells in H(2)O(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.

[Effect of silent information regulator 1 on the LPS induced lncRNA expression of macrophages in mice].

Objective: To investigate the effect of Silent information regulator 1 (Sirt1) on the expression profile of long non-coding RNA (lncRNA) in macrophages upon lipopolysaccharide (LPS) treatment. Methods: Peritoneal macrophages (PM) were isolated from nine wild-type C57BL/6 male mice (wild-type group) and nine myeloid-specific Sirt1 knock-out mice (knock-out group). RNA samples were extracted from macrophages stimulated with 1 µg/ml LPS. Sequencing and the differentially expressed lncRNA were screened after the RNA was quantified. The threshold set for up-and down-regulated genes was a fold change (wild-type group/knock-out group) ≥2 and P≤0.05. Afterwards, gene ontology (GO) and pathway enrichment analysis were conducted and co-expression network map was constructed. Results: Four hundred and forty five lncRNA genes were differentially expressed (185 lncRNA genes were up-regulated and 260 lncRNA genes were down-regulated). Two hundred mRNA genes were differentially expressed (113 mRNA genes were up-regulated and 87 mRNA genes were down-regulated). It was found that the differentially expressed lncRNA genes and the predicted corresponding target genes were mainly distributed in the regions of biological processes of macrophage inflammatory response, macrophage chemotaxis and cell metabolism by GO and pathway enrichment analysis. Conclusion: lncRNA expression profile changes significantly in LPS induced macrophages isolated from Sirt1 knock out mice, which is closely related to the function of macrophages.

[The role of facial nerve motor evoked potentials in predicting facial nerve function in vestibular schwannoma surgery].

Objective: To analyze the role of facial nerve motor evoked potentials in predicting facial nerve function in vestibular schwannoma surgery. Methods: In a retrospective clinical study of 226 patients with acoustic neuroma, admitted to our hospital from January 2016 to May 2019, were investigated by facial nerve motor evoked potentials (FNMEP) elicited by multi-pulse transcranial electrical motor cortex stimulation from. For recording the same electrode set-up was used as for continuous EMG monitoring of the orbicularis oculi,oris muscles and mentalis. Pre-surgical (opening dural), intraoperative and post-surgical (closing dural) FNMEP amplitudes and latencies were recorded. End (closing dura) to start (opening dura) amplitude ratios were compared to early-term(3 day after surgery) facial nerve function by House-Brackmann(HB) Grading. Results: 201 patients(88.9%) get a total tumor resection, 15 patients (6.6%) were a subtotal resection, 10 patients(4.4%) were a partial resection. 100 percent of patients had a integrated anatomical preservation of facial nerves, there were four (1.8%) death cases in this group. Reliable FNMEPs were obtained in all patients. The ratio of end-operative to start-operative FNMEP-amplitude showed a negative correlation with early facial nerve function. Correlation was especially close with early function: an amplitude preservation rate of 85.3% led to HB Ⅰ or Ⅱ in 190(84.1%) patients, of 45.6% to HB Ⅲ in 17(7.5%) patients, of 23.1% to HB Ⅳ in 13(5.8%) patients and of 6.7% to HB Ⅴor Ⅵ in 6(2.7%) patients. There was a negative correlation between FNMEP amplitude ratio and post-surgical early HB grading(r=-0.895, P=0.000). Conclusion: FNMEP was highly reliable in predicting early postoperative facial function of the resection of vestibular schwannoma, was a valid protection technique of facial nerve.

Effect of IL-1ß on apoptosis of synovial cells in rheumatoid arthritis rats via the NF-κB pathway.

OBJECTIVE: The aim of this study was to investigate the role of interleukin-1ß (IL-1ß) in the apoptosis of synovial cells in rheumatoid arthritis (RA) rats, and to explore the underlying mechanism. MATERIALS AND METHODS: The apoptosis of the synovial cells in RA rats in the IL-1ß group and the control group was analyzed by scoring under an electron microscope. The expressions of cleaved-poly (ADP-ribose) polymerase (PARP), PARP and anti-apoptosis gene products in synovial cells of IL-1ß treated RA rats were explored as well. Meanwhile, the expressions of B-cell lymphoma 2 (Bcl-2), Bcl-xL, and Active-Caspase3 in the synovial cells of RA rats with IL-1ß treatment were evaluated by the Western blotting. To further clarify the relationship between IL-1ß and the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway in the synovial cells of RA rats, the expressions of NF-κB regulated the gene products of matrix metalloproteinase-3 (MMP-3), MMP-9, cyclooxygenase-2 (Cox-2), and vascular endothelial growth factor (VEGF) in synovial cells of RA rats after that we investigated the treatment with IL-1ß (was investigated). In addition, the expression of NF-κB in the synovial cells of RA rats treated with IL-1ß was determined. RESULTS: The results showed that, compared with the control group, IL-1ß treatment significantly increased the number of apoptotic cells. This meant that IL-1ß treatment could promote the apoptosis of the synovial cells (p<0.05). IL-1ß treatment significantly promoted the expression level of cleaved-PARP (p<0.05). However, it remarkably reduced the expressions of Bcl-2 and Bcl-xL (p<0.05). Meanwhile, the level of the active-Caspase3 in the synovial cells of RA rats treated with IL-1ß was significantly enhanced (p<0.01). In comparison with the control group, the IL-1ß group exhibited significantly elevated expressions of NF-κB-regulated gene products in the synovial cells of RA rats (p<0.01). Besides, the positive markers of the activated NF-κB were detected in the synovial cells of RA rats in the IL-1ß group and the control group. The results demonstrated that they were mainly located in the nucleus of the IL-1ß group. CONCLUSIONS: IL-1ß can promote the apoptosis of the synovial cells in RA rats via the NF-κB pathway.