Objective To explore the molecular mechanism of N6-methyladenosine(m6A)methylation mediated by methyltransferase 3(METTL3)in regulating lipopolysaccharide(LPS)-induced endothelial cell permeability changes.Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro.HUVECs were treated with LPS 50,125,250,500,1 000,2 000 ng/ml for 24 h.METTL3 mRNA expression was detected by Real-time PCR.After HUVECs were intervened with 500 ng/ml for 24 h,the methylation level of m6A was detec-ted,and cell permeability was measured by cell permeability test.Real-time PCR and Western blot were used to detect mRNA and protein expression of intercellular junction proteins(Claudin-5,Occludin and VE-caherin).METTL3 overexpressed stable cell lines were constructed to measure the changes of m6A methylation level and per-meability of endothelial cells during METTL3 overexpression.Results Compared to the control group,LPS inhibi-ted the expression of HUVECs METTL3 mRNA,decreased the methylation of m6A,increased the cell permeabili-ty,and decreased the mRNA and protein expression of intercellular junction proteins(Claudin-5,Occludin and VE-Caherin).When METTL3 was overexpressed,the m6A methylation levels of endothelial cells were enhanced,and the increase of endothelial cell permeability induced by LPS was reversed.Conclusion METTL3-mediated m6A methylation can improve the permeability of endothelial cells induced by sepsis.

Objective : To investigate the molecular mechanism of Toll⁃like receptor 4 ( TLR4)/Ras homologue A (RhoA) signaling pathway involved in regulating the effect of septic serum on vascular endothelial cell permeability before and after continuous hemofiltration. Methods : The serum of 5 patients with sepsis before and after continuous hemofiltration treatment was collected , and the levels of inflammatory cytokines in serum before and after hemofiltration were detected. Human umbilical vein endothelial cells (HUVEC) were treated with serum before and after continuous hemofiltration for 24 hours. The expression of VE⁃cadherin , F ⁃actin , TLR4 and RhoA in vascular endothelial cells were detected by Western blot. A TLR4 low expression cell line was constructed to detect the effect of TLR4 low expression on the expression of VE⁃cadherin , F ⁃actin and RhoA and the permeability of endothelial cells. Results : After continuous blood treatment , the serum levels of TLR4 , RhoA , interleukin⁃1 (IL⁃1) , interleukin⁃6 (IL⁃6) and tumor necrosis factor⁃α (TNF⁃α ) significantly decreased. The expression levels of VE⁃cadherin , F ⁃actin , TLR4 and RhoA in the serum intervention group after continuous hemofiltration treatment significantly decreased , and the cell permeability significantly decreased. Low expression of TLR4 significantly promoted the expression of VE⁃cadherin and F ⁃actin , and inhibited the expression of RhoA protein. Conclusion TLR4/RhoA signaling pathway is involved in the regulation of changes in vascular endothelial cell permeability induced by septic serum after continuous hemofiltration treatment.

Objective:To investigate the application value of domestic robotic surgical system in radical resection of hilar cholangiocarcinoma (hCCA).Methods:The retrospective and descriptive study was constructed. The clinicopathological data of a 66-year-old male patient with hCCA who was admitted to the Second Hospital of Shandong University in November 2022 were collected. Radical resection of hCCA was performed using the Toumai? laparoscopic surgical robot system. Observation indicators: (1) intraoperative conditions; (2) postoperative conditions; (3) follow-up. Count data were represented as absolute numbers and (or) percentages.Results:(1) Intraoperative conditions. The patient underwent radical resection of hCCA successfully using robotic surgical system, including tumor resection, lymph node dissection, and gastrointestinal reconstruc-tion. The operation time and volume of intraoperative blood loss were 400 minutes and 100 mL, respectively, and no intraoperative blood transfusion was required. (2) Postoperative conditions. The patient began ambulation on postoperative day 1 and began taking liquid food on postoperative day 4. Liver function examination and abdominal computed tomograph (CT) on postoperative day 5 showed a decrease in serum bilirubin, no biliary or intestinal leakage, and no edema or necrosis at the anastomotic site. The abdominal pre-anastomotic and post-anastomotic drainage tubes were removed on the 7th and 9th day after the surgery respectively, and the patient was discharged on the 10th day after surgery. Results of postoperative pathological examination showed moderately to poorly differentiated adenocarcinoma of the bile duct. Immunostaining was positive for CK7 and CK19. The Ki-67 proliferation index was 35%. The proximal and distal resection margin of bile duct were negative. The tumor diameter was 30 mm. Perineural invasion was positive. The surgical specimen margin was negative. Two lymph nodes were identified containing tumor cells positive for malignancy. No tumor cell metastasis was detected in the No.8, No.12 or gastric lesser curvature lymph nodes submitted for pathological examination. (3) Follow-up. The patient was followed up at postoperative 1-, 3-, 5-month after discharge. During follow-up period, results of liver function examination and abdominal CT showed liver function restore to normal levels, no complication such as biliary fistula, intestinal fistula, gastroparesis or tumor metastasis.Conclusion:The Domestic Toumai ? laparoscopic surgical robot system can be applied to radical resection of hCCA.

Objective: To investigate the influencing factors of depressive symptoms among rural elderly patients with chronic diseases in China, so as to provide insights into depression prevention and control among the rural elderly patients with chronic diseases. Methods: The basic demographics, health status, and lifestyle of rural residents at ages of 65 years and older who had at least one chronic disease were retrieved from The Chinese Longitudinal Healthy Longevity Survey (CLHLS) database in 2018, and participants' depressive symptoms were assessed with The Center for Epidemiological Studies Depression-10 (CES-D-10) scale. Factors affecting the depressive symptoms were identified with a multivariable logistic regression model. Results: Totally 5 146 rural elderly patients with chronic diseases were enrolled, including 2 373 men (46.11%) and 2 773 women (53.89%). The prevalence of depressive symptoms was 27.13%. Multivariable logistic regression analysis identified having two and more children (OR=0.614, 95%CI: 0.387-0.975), living alone (OR=1.450, 95%CI: 1.192-1.764), life satisfaction (general, OR=1.933, 95%CI: 1.651-2.264; low, OR=5.366, 95%CI: 3.488-8.254), self-assessed health status (general, OR=2.697, 95%CI: 2.284-3.185; poor, OR=5.338, 95%CI: 4.262-6.685), disability in instrumental activities of daily living (OR=1.592, 95%CI: 1.328-1.908), sleep duration (normal, OR=0.502, 95%CI: 0.429-0.586; too long, OR=0.494, 95%CI: 0.405-0.603), exercise (OR=0.721, 95%CI: 0.607-0.856), watching TV (OR=0.787, 95%CI: 0.664-0.933), and gardening activities (OR=0.781, 95%CI: 0.626-0.975) as factors affecting depressive symptoms among rural elderly patients with chronic diseases. Conclusions The prevalence of depressive symptoms was 27.13% among rural elderly patients with chronic diseases. Number of children, living style, life satisfaction, health status, sleep duration, exercise, watching TV, and gardening activities are associated with the development of depressive symptoms among rural elderly patients with chronic diseases.

Objective:To compare the effect of three-dimensional CT angiography (CTA) and color Doppler ultrasound (CDU) in reconstruction of soft tissue defects after oral cancer surgery with anterolateral thigh flap (ALTF).Methods:Patients who underwent reconstruction of postoperative defects with ALTF in Department of Oral and Maxillofacial Surgery, the Affiliated Stomatological Hospital of Southwest Medical University from January 2013 to January 2020 were retrospectively reviewed. According to the imaging method used for examining perforating artery, all patients were divided into two groups. In group A, CTA and three-dimensional vascular reconstruction were used to locate the lateral femoral circumflex artery and its perforating vessels, and the individualized ALTF was designed. In group B, the lateral circumflex femoral artery and its perforating vessels were located by CDU. The clinical features, operation time, flap survival rate, postoperative complication rate, image quality of angiography, difference of superficial skin points of perforating vessels before and during operation, and diameter of perforators were compared between the two groups. The measurement data were expressed by Mean±SD, and the differences between the two groups were compared by independent sample t-test; paired sample t-test was used to compare the intra-group differences; patients’ clinical features and other enumeration data were expressed in the number of cases (%), and analyzed by chi-square test; reliability analysis was adopted for the image quality score of two doctors at different times, taking Cronbach’s α value; correlations between body mass index (BMI) and flap survival rate were tested by Pearson correlation, whereas the correlation between flap survival rate and underlying diseases, drug combination, bad smoking and drinking habits, and surgical complications were tested by Spearman correlation, which were all performed using SPSS version 20.0 at significance level P<0.05. Results:A total of 50 patients with oral cancer were collected, 25 patients in each group. There were no significant differences between the two groups in clinical features such as sex distribution, age, TNM stage, BMI, underlying diseases, drug combination, bad smoking and drinking habits, tumor location and so on ( P>0.05). The mean operation time of group A was significantly shorter than that of group B[(67.64±5.94) min vs. (70.88±4.88) min, P<0.05]. All flaps in group A survived; one case in group B had complete flap necrosis. There was no significant difference in flap survival rate between the two groups(100% vs. 96%, P>0.05). One case in group A and two cases in group B had complications such as effusion of donor site wound, and there was no significant difference ( P>0.05). Compared with the image quality of the two groups, the angiographic quality score of group A was significantly higher than that of group B, the difference was statistically significant [(3.08±0.64) scores vs. (2.56±0.65) scores, P<0.05]. In group A, the distance difference between the position of the skin superficial point of perforator vessel positioned before operation and during operation was significantly shorter than that in group B, and the difference was statistically significant [(1.32±0.50) mm vs. (1.75±0.84) mm, P<0.05]. In group A, the diameter of perforating artery measured by imaging before operation was (1.68±0.17) mm, which had no significant difference with the actual value[(1.70±0.18) mm] ( P>0.05); whereas, in group B, the diameter of perforating artery measured by imaging before operation was (1.77±0.14) mm. The actual measured value during operation was (1.66±0.15) mm, the difference was statistically significant ( P<0.05). A significant correlation emerged between surgical complications and flap survival rate ( r=0.57, P<0.001), however, there were no significant correlations between BMI, combined with basic diseases, combined medication, bad smoking and drinking habits and flap survival ( P>0.05). Conclusions:Compared with CDU, CTA combined with three-dimensional vascular reconstruction is a feasible and reliable method for preoperative evaluation, flap design. The flap preparation time is shorter. Therefore, it is a good method of preoperative vascular localization.

Objective:To compare the effect of three-dimensional CT angiography (CTA) and color Doppler ultrasound (CDU) in reconstruction of soft tissue defects after oral cancer surgery with anterolateral thigh flap (ALTF).Methods:Patients who underwent reconstruction of postoperative defects with ALTF in Department of Oral and Maxillofacial Surgery, the Affiliated Stomatological Hospital of Southwest Medical University from January 2013 to January 2020 were retrospectively reviewed. According to the imaging method used for examining perforating artery, all patients were divided into two groups. In group A, CTA and three-dimensional vascular reconstruction were used to locate the lateral femoral circumflex artery and its perforating vessels, and the individualized ALTF was designed. In group B, the lateral circumflex femoral artery and its perforating vessels were located by CDU. The clinical features, operation time, flap survival rate, postoperative complication rate, image quality of angiography, difference of superficial skin points of perforating vessels before and during operation, and diameter of perforators were compared between the two groups. The measurement data were expressed by Mean±SD, and the differences between the two groups were compared by independent sample t-test; paired sample t-test was used to compare the intra-group differences; patients’ clinical features and other enumeration data were expressed in the number of cases (%), and analyzed by chi-square test; reliability analysis was adopted for the image quality score of two doctors at different times, taking Cronbach’s α value; correlations between body mass index (BMI) and flap survival rate were tested by Pearson correlation, whereas the correlation between flap survival rate and underlying diseases, drug combination, bad smoking and drinking habits, and surgical complications were tested by Spearman correlation, which were all performed using SPSS version 20.0 at significance level P<0.05. Results:A total of 50 patients with oral cancer were collected, 25 patients in each group. There were no significant differences between the two groups in clinical features such as sex distribution, age, TNM stage, BMI, underlying diseases, drug combination, bad smoking and drinking habits, tumor location and so on ( P>0.05). The mean operation time of group A was significantly shorter than that of group B[(67.64±5.94) min vs. (70.88±4.88) min, P<0.05]. All flaps in group A survived; one case in group B had complete flap necrosis. There was no significant difference in flap survival rate between the two groups(100% vs. 96%, P>0.05). One case in group A and two cases in group B had complications such as effusion of donor site wound, and there was no significant difference ( P>0.05). Compared with the image quality of the two groups, the angiographic quality score of group A was significantly higher than that of group B, the difference was statistically significant [(3.08±0.64) scores vs. (2.56±0.65) scores, P<0.05]. In group A, the distance difference between the position of the skin superficial point of perforator vessel positioned before operation and during operation was significantly shorter than that in group B, and the difference was statistically significant [(1.32±0.50) mm vs. (1.75±0.84) mm, P<0.05]. In group A, the diameter of perforating artery measured by imaging before operation was (1.68±0.17) mm, which had no significant difference with the actual value[(1.70±0.18) mm] ( P>0.05); whereas, in group B, the diameter of perforating artery measured by imaging before operation was (1.77±0.14) mm. The actual measured value during operation was (1.66±0.15) mm, the difference was statistically significant ( P<0.05). A significant correlation emerged between surgical complications and flap survival rate ( r=0.57, P<0.001), however, there were no significant correlations between BMI, combined with basic diseases, combined medication, bad smoking and drinking habits and flap survival ( P>0.05). Conclusions:Compared with CDU, CTA combined with three-dimensional vascular reconstruction is a feasible and reliable method for preoperative evaluation, flap design. The flap preparation time is shorter. Therefore, it is a good method of preoperative vascular localization.

Objective:To explore the risk factors of acute renal injury (AKI) in exertional heat radiation disease (EHS) .Methods:In november 2019, the clinical data of 69 EHS patients admitted from July 2015 to September 2019 were reviewed. The general data, laboratory indexes, Glasgow score (GCS) at admission, 24-hour acute physiology and chronic health score Ⅱ (APACHE Ⅱ) , exposure time rate and physical labor intensity were collected. According to the occurrence of AKI, the patients were divided into AKI group and non-AKI group, 31 and 38 in each group. The differences of general data and laboratory indexes between the two groups were compared, and the t and Mann-Whitney U test were used to compare the two groups. The enumeration data are expressed by examples and constituent ratio (%) . Independent sample χ 2 test is used for inter-group comparison, and multiple test is used for multi-sample comparison. The correlation was analyzed by linear regression. Risk factors were analyzed by Logistic regression analysis. Results:At discharge, 31 of 69 EHS patients developed AKI. Compared with the non-AKI group, the heart rate, white blood cell count, lactic acid, D-dimer and myoglobin were higher; MAP, platelet count and PH were lower in the AKI group. The difference was statistically significant ( P<0.05) . APACHE Ⅱ score, core temperature, time to drop to 38.5 ℃, contact time rate, platelet count, pH, lactic acid, D-dimer and myoglobin were all correlated with creatinine ( r=0.57, 0.42, 0.80, 0.78, 0.57, 0.43, 0.51, 0.55, 0.79) . APACHE Ⅱ score, time to drop to 38.5C, Lac and MYO are the risk factors of AKI in EHS patients. Multivariate Logistic regression analysis showed that the time required to drop to 38.5C was an independent risk factor for the occurrence of AKI. Conclusion:AKI is a serious complication of EHS. EHS complicated with AKI, should be identified early and effective intervention measures should be taken.

Objective:To explore the risk factors of acute renal injury (AKI) in exertional heat radiation disease (EHS) .Methods:In november 2019, the clinical data of 69 EHS patients admitted from July 2015 to September 2019 were reviewed. The general data, laboratory indexes, Glasgow score (GCS) at admission, 24-hour acute physiology and chronic health score Ⅱ (APACHE Ⅱ) , exposure time rate and physical labor intensity were collected. According to the occurrence of AKI, the patients were divided into AKI group and non-AKI group, 31 and 38 in each group. The differences of general data and laboratory indexes between the two groups were compared, and the t and Mann-Whitney U test were used to compare the two groups. The enumeration data are expressed by examples and constituent ratio (%) . Independent sample χ 2 test is used for inter-group comparison, and multiple test is used for multi-sample comparison. The correlation was analyzed by linear regression. Risk factors were analyzed by Logistic regression analysis. Results:At discharge, 31 of 69 EHS patients developed AKI. Compared with the non-AKI group, the heart rate, white blood cell count, lactic acid, D-dimer and myoglobin were higher; MAP, platelet count and PH were lower in the AKI group. The difference was statistically significant ( P<0.05) . APACHE Ⅱ score, core temperature, time to drop to 38.5 ℃, contact time rate, platelet count, pH, lactic acid, D-dimer and myoglobin were all correlated with creatinine ( r=0.57, 0.42, 0.80, 0.78, 0.57, 0.43, 0.51, 0.55, 0.79) . APACHE Ⅱ score, time to drop to 38.5C, Lac and MYO are the risk factors of AKI in EHS patients. Multivariate Logistic regression analysis showed that the time required to drop to 38.5C was an independent risk factor for the occurrence of AKI. Conclusion:AKI is a serious complication of EHS. EHS complicated with AKI, should be identified early and effective intervention measures should be taken.

Objective:To compare the differences of short and long-term outcomes between laparoscopic surgery and open surgery treatment of intrahepatic cholangiocarcinoma patients.Methods:A retrospective cohort study was conducted to collect the clinical data of 118 patients with intrahepatic cholangiocarcinoma who underwent surgery in Qilu Hospital of Shandong University from January 2015 to June 2020. They were divided into laparoscopy group and open group according to the operation methods. The perioperative data, such as intraoperative surgical conditions, hospital costs, postoperative complications, postoperative blood biochemical tests, and the follow-up data of the two groups were compared.Results:In the laparoscopic group, there were 40 patients, 18 males and 22 females, aged (61.5±9.1) years. There were 78 patients in the open group, 48 males and 30 females, aged (61.2±8.3) years. The tumor size of the laparoscopic group was (4.4±1.8) cm, which was smaller than that of the open group (6.0±3.3) cm, and the differences were statistically significant ( P<0.05). In the laparoscopic group, 4 cases (10%) were converted to open surgery. The intraoperative blood loss, intraoperative blood transfusion proportion, 3 or more liver segments resection proportion and hospital costs of laparoscopic group were lower than those of open group [200.0(100.0, 261.8) ml vs. 300.0(100.0, 400.0) ml, 5.0%(2/40) vs. 26.9%(21/78), 37.5%(15/40) vs. 66.7%(52/78), (6.2±2.0) wan yuan vs. (7.2±2.3) wan yuan], the differences were statistically significant (all P<0.05). There were no significant differences in the incidence of postoperative complications between the two groups ( P>0.05). On the first post-operative day, ALT serum level and the third post-operative day TBil serum level in the laparoscopic group were lower than those in the open group [188.5(130.5, 274.0) U/L vs. 320.0(144.0, 427.0) U/L, 26.4(18.3, 26.4) μmol/L vs. 31.6(18.8, 37.5) μmol/l], the differences were statistically significant ( P<0.05). There were no significant differences in 1-year and 2-year overall survival rate and disease-free survival rate between the two groups ( P>0.05). Conclusion:Compared with open surgery, laparoscopic surgery in the treatment of intrahepatic cholangiocarcinoma has better short-term outcomes, and can achieve similar results in medium- or long-term outcomes.

Objective:To evaluate the prognostic value of different critical care scoring systems in 28-day survival rate of patients with heat stroke.Methods:A retrospective analysis was conducted on the clinical data of 71 patients with heat stroke admitted to the department of emergency medicine of Beijing Luhe Hospital. Capital Medical University from July 2015 to September 2018. The general information and the worst values of vital signs and related pathophysiological indicators within 24 hours were collected and the sequential organ failure assessment (SOFA) , multiple organ dysfunction (MODS) , simplified acute physiological scoreⅡ (SAPS Ⅱ) and acute physiology and chronic health evaluationⅡ (APACHE Ⅱ) were calculated. The patients were divided into the survival group ( n=45) and the non-survival group ( n=26) according to 28-day prognosis, and the clinical data and scores of the two groups were compared.The ROC curve was drawn to analyze the evaluation value of each scoring system on the survival rate of patients at 28-day. Kaplan-Meier method was used to plot the survival curve of patients. Results:There were no significant differences in age, sex, vital signs and laboratory parameters between two groups ( P>0.05) . In non-survival patients, SOFA, SAPS Ⅱ, APACHE Ⅱ scores were significantly elevated in the survival group ( P<0.05) . ROC curve analysis showed that the area under ROC curve (AUC) of SOFA score for predicting 28-day survival rate was the highest, which was significantly higher than the APACHE Ⅱ, SAPS Ⅱ, MODS score. When the best cut-off value of SOFA score was 9.0, the sensitivity was 84.6%, and the specificity was 71.1%. Kaplan-Meier survival analysis showed that 28-day survival rate after hospital discharge in patients with SOFA score<9 ( n=27) was significantly higher than that in patients with SOFA score ≥9.0 (χ 2=1.0, P<0.01) . Conclusion:SOFA, APACHE Ⅱ, SAPS Ⅱ on admission have been proved to have good prognostic ability to predict 28-day prognosis in heat stroke patients. Among them, SOFA score system has more accurate prediction value.