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 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 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 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 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 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 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 evaluate the clinical value of holographic image navigation in urological laparoscopic and robotic surgery.Methods The data of patients were reviewed retrospectively for whom accepted holographic image navigation laparoscopic and robotic surgery from Jan.2019 to Dec.2019 in Beijing United Family Hospital and other 18 medical centers,including 78 cases of renal tumor,2 cases of bladder cancer,2 cases of adrenal gland tumor,1 cases of renal cyst,1 case of prostate cancer,1 case of sweat gland carcinoma with lymph node metastasis,1 case of pelvic metastasis after radical cystectomy.All the patients underwent operations.In the laparoscopic surgery group,there were 27 cases of partial nephrectomy,1 case of radical prostatectomy,2 cases of radical cystectomy and 2 cases of adrenalectomy.In the da Vinci robotic surgery group of 54 cases,there were 51 cases of partial nephrectomy,1 case of retroperitoneal lymph node dissection,1 case of retroperitoneal bilateral renal cyst deroofing and 1 case of resection of pelvic metastasis.There were 41 partial nephrectomy patients with available clinical data for statistic,with a median age of 53.5 years (range 24-76),including 26 males and 15 females.The median R.E.N.A.L score was 7.8 (range 4-11).Before the operation,the engineers established the holographic image based on the contrast CT images and reports.The surgeon applied the holographic image for preoperative planning.During the operation,the navigation was achieved by real time fusing holographic images with the laparoscopic surgery images in the screen.Results All the procedures had been complete uneventfully.The holographic images helped surgeon in understanding the visual three-dimension structure and relation of vessels supplying tumor or resection tissue,lymph nodes and nerves.By manipulating the holographic images extracorporeally,the fused image guide surgeons about location vessel,lymph node and other important structure and then facilitate the delicate dissection.For the 41 cases with available clinical data including 23 cases of robotic-assisted partial nephrectomy and 18 cases of laparoscopic nephrectomy,the median operation time was 140 (range 50-225) min,the median warm ischemia time was 23 (range 14-60) min,the median blood loss was 80(range 5-1 200) ml.In the robotic surgery group,the median operation time was 140 (range 50-215)min,the median warm i schemia time was 21 (range 17-40)min,the median blood loss was 150(range 30-1 200)ml.In the laparoscopic surgery group,the median operation time was 160(range 80-225)min,the median warm ischemia time was 25 (range 14-60)min,the median blood loss was 50 (range 5-1 200) ml.All the patients had no adjacent organ injury during operation.There were 2 cases with Clavien Ⅱ complications.One required transfusion and the other one suffered hematoma post-operation.However,the tumors were located in the renal hilus for these 2 cases and the R.E.N.A.L scores were both 11.Conclusions Holographic image navigation can help location and recognize important anatomic structures during the surgical procedures..This technique will reduce the tissue injury,decrease the complications and improve the success rate of surgery.