ObjectiveTo investigate the therapeutic effect of paeoniflorin on metabolic dysfunction-associated fatty liver disease （MASLD） combined with insulin resistance （IR）， and to elucidate its regulatory mechanisms on the Toll-like receptor 4 （TLR4）/nuclear transcription factor （NF）-κB-insulin receptor substrate 1 （IRS1） inflammation-insulin signaling axis. MethodsMASLD mouse model was established using a 60% high-fat diet. Mice were randomly assigned to a normal control group， a MASLD model group， and paeoniflorin groups （low dose： 25 mg·kg^-1·d^-1， medium dose： 50 mg·kg^-1·d^-1， high dose： 100 mg·kg^-1·d^-1）， receiving intragastric administration for consecutive 12 weeks. Concurrently， a palmitic acid/oleic acid （PA/OA）-induced lipid deposition model was established in HepG2 cells. Cell counting kit-8 （CCK-8） assay was performed to determine the optimal concentrations of PA/OA and paeoniflorin for intervention. Mice body weight， liver index， and serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， triglycerides （TG）， and total cholesterol （TC） were measured. Fasting blood glucose （FBG） and fasting insulin （FINS） levels were measured， and the homeostatic model assessment of insulin resistance （HOMA-IR） was calculated， while an oral glucose tolerance test （OGTT） was performed to evaluate glucose metabolism. Hematoxylin and eosin （HE） staining was used to observe pathological changes and inflammatory cell infiltration in liver tissue， and oil red O staining was used to evaluate lipid droplet deposition in liver tissue and HepG2 cells. Potential targets were screened using network pharmacology and molecular docking. Western blot analysis was performed to detect the expression of HSP90AA1， TLR4， p-NF-κB p65， p-IRS1， and p-Akt proteins in liver tissue and HepG2 cells. ResultsCompared with the normal control group， mice in the MASLD model group exhibited significantly increased body weight， liver index， and serum levels of ALT， AST， TG， and TC， as well as markedly elevated FBG， FINS， and HOMA-IR， and impaired glucose tolerance （P<0.01）. HE staining revealed marked hepatic steatosis and inflammatory cell infiltration， while oil red O staining showed a significant increase in lipid droplet deposition. Compared with the MASLD model group， after intervention with various doses of paeoniflorin， the increase in body weight and liver index was reduced， serum ALT， AST， TG， and TC levels decreased， and FBG， FINS， and HOMA-IR were significantly lowered， with glucose tolerance markedly improved （P<0.05， P<0.01）， showing a certain dose-dependent trend. HE and oil red O staining results showed a marked reduction in hepatic steatosis and lipid deposition. In vitro experiments demonstrated that， compared with the control group， PA/OA treatment significantly induced increased lipid deposition in HepG2 cells. Compared with the model group， paeoniflorin intervention significantly reduced intracellular lipid droplets， and lipid deposition showed a dose-dependent decreasing trend. Mechanism studies indicated that， compared with the normal control group， the MASLD model group exhibited significantly elevated expression of TLR4， HSP90AA1， and p-NF-κB p65， while p-IRS1 （Ser307） expression was elevated and p-Akt （Ser473） expression was reduced （P<0.01）. Compared with the model group， the expression of the aforementioned proteins was significantly reversed in all treatment groups following intervention （P<0.05， P<0.01）. ConclusionPaeoniflorin significantly alleviates lipid deposition and insulin resistance in MASLD mice. Its mechanism of action may involve targeting HSP90AA1， TLR4， and NF-κB1 to regulate the TLR4/NF-κB-IRS1 inflammation-insulin signaling axis. This regulation suppresses inflammatory responses and restores insulin signaling， thereby ameliorating abnormalities in glucose and lipid metabolism.

ObjectiveTo investigate the therapeutic effect of paeoniflorin on metabolic dysfunction-associated fatty liver disease （MASLD） combined with insulin resistance （IR）， and to elucidate its regulatory mechanisms on the Toll-like receptor 4 （TLR4）/nuclear transcription factor （NF）-κB-insulin receptor substrate 1 （IRS1） inflammation-insulin signaling axis. MethodsMASLD mouse model was established using a 60% high-fat diet. Mice were randomly assigned to a normal control group， a MASLD model group， and paeoniflorin groups （low dose： 25 mg·kg^-1·d^-1， medium dose： 50 mg·kg^-1·d^-1， high dose： 100 mg·kg^-1·d^-1）， receiving intragastric administration for consecutive 12 weeks. Concurrently， a palmitic acid/oleic acid （PA/OA）-induced lipid deposition model was established in HepG2 cells. Cell counting kit-8 （CCK-8） assay was performed to determine the optimal concentrations of PA/OA and paeoniflorin for intervention. Mice body weight， liver index， and serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， triglycerides （TG）， and total cholesterol （TC） were measured. Fasting blood glucose （FBG） and fasting insulin （FINS） levels were measured， and the homeostatic model assessment of insulin resistance （HOMA-IR） was calculated， while an oral glucose tolerance test （OGTT） was performed to evaluate glucose metabolism. Hematoxylin and eosin （HE） staining was used to observe pathological changes and inflammatory cell infiltration in liver tissue， and oil red O staining was used to evaluate lipid droplet deposition in liver tissue and HepG2 cells. Potential targets were screened using network pharmacology and molecular docking. Western blot analysis was performed to detect the expression of HSP90AA1， TLR4， p-NF-κB p65， p-IRS1， and p-Akt proteins in liver tissue and HepG2 cells. ResultsCompared with the normal control group， mice in the MASLD model group exhibited significantly increased body weight， liver index， and serum levels of ALT， AST， TG， and TC， as well as markedly elevated FBG， FINS， and HOMA-IR， and impaired glucose tolerance （P<0.01）. HE staining revealed marked hepatic steatosis and inflammatory cell infiltration， while oil red O staining showed a significant increase in lipid droplet deposition. Compared with the MASLD model group， after intervention with various doses of paeoniflorin， the increase in body weight and liver index was reduced， serum ALT， AST， TG， and TC levels decreased， and FBG， FINS， and HOMA-IR were significantly lowered， with glucose tolerance markedly improved （P<0.05， P<0.01）， showing a certain dose-dependent trend. HE and oil red O staining results showed a marked reduction in hepatic steatosis and lipid deposition. In vitro experiments demonstrated that， compared with the control group， PA/OA treatment significantly induced increased lipid deposition in HepG2 cells. Compared with the model group， paeoniflorin intervention significantly reduced intracellular lipid droplets， and lipid deposition showed a dose-dependent decreasing trend. Mechanism studies indicated that， compared with the normal control group， the MASLD model group exhibited significantly elevated expression of TLR4， HSP90AA1， and p-NF-κB p65， while p-IRS1 （Ser307） expression was elevated and p-Akt （Ser473） expression was reduced （P<0.01）. Compared with the model group， the expression of the aforementioned proteins was significantly reversed in all treatment groups following intervention （P<0.05， P<0.01）. ConclusionPaeoniflorin significantly alleviates lipid deposition and insulin resistance in MASLD mice. Its mechanism of action may involve targeting HSP90AA1， TLR4， and NF-κB1 to regulate the TLR4/NF-κB-IRS1 inflammation-insulin signaling axis. This regulation suppresses inflammatory responses and restores insulin signaling， thereby ameliorating abnormalities in glucose and lipid metabolism.

Objective:To analyze the histopathological characteristics of peri-implant soft tissue in re-constructed jaws and the changes after keratinized mucosa augmentation(KM A)with free gingival graft(FGG).Methods:Twenty patients were enrolled in this study.Five patients of them,who were perio-dontal and systemic healthy and referred for crown lengthening before restoration with healthy keratinized gingiva collected were enrolled as healthy controls.15 patients of them were with fibula or iliac bone flaps jaw reconstruction(10 with fibula flap and 5 with iliac flap),who were referred to FGG and implant exposures before restoration.Soft tissue was collected before FGG in reconstructed jaws,and in 5 patients(3 with fibula flap and 2 with iliac flap)8 weeks after FGG if a second surgery was conducted.Histologi-cal analysis with hematoxylin-eosin stain and immunological analysis to interlukin-1(IL-1),interlukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were performed.Results:Thickness from the bottom of stratum basale to the top of stratum granulosum and thickness of keratinized layer in reconstructed jaws were significantly lower compared with that of natural healthy keratinized gingiva[0.27(0.20,0.30)mmvs.0.36(0.35,0.47)mm,P＜0.05;16.49(14.90,23.37)μm vs.26.37(24.12,31.53)μm,P＜0.05].In the reconstructed area,thickness from the bottom of stratum basale to the top of stra-tum granulosum increased after KMA with FGG[0.19(0.16,0.25)mm vs.0.38(0.25,0.39)mm,P=0.059]and the thickness of keratinized layer significantly increased after KMA with FGG[16.42(14.16,22.35)μm vs.28.57(27.16,29.14)μm,P＜0.05],which was similar to that in the con-trol group.Furthermore,the number of positive cells of IL-1,IL-6 and TNF-α significantly increased after KMA[0.67(0.17,8.93)vs.11.00(9.16,18.00);13.00(8.50,14.14)vs.21.89(15.00,28.12);0.22(0.04,0.63)vs.2.83(1.68,5.00),respectively,P＜0.05]as well as the average optical density value[0.15(0.14,0.17)vs.0.18(0.17,0.21);0.28(0.26,0.33)vs.0.36(0.33,0.37);0.23(0.22,0.29)vs.0.30(0.28,0.42),respectively,P＜0.05],which was similar to that in the healthy keratinized gingiva.Conclusion:The lack of rete pegs and inflammato-ry factors were common in soft tissue with jaw reconstruction.FGG can improve the quality of the epithe-lium and may improve the stability of the mucosa around implants.

Objective:To analyze the concentration of formic acid,propionic acid and butyric acid in gingival crevicular fluid(GCF)of patients with stages Ⅲ and Ⅳ periodontitis,and their relationship with periodontitis.Methods:The study enrolled 37 systemically healthy patients with periodontitis and 19 healthy controls who visited Department of Periodontology,Peking University School and Hospital of Sto-matology from February 2008 to May 2011.Their GCFs were collected from the mesial-buccal site of one molar or incisor in each quadrant.Periodontal clinical parameters,including plaque index(PLI),probing depth(PD),bleeding index(BI),and attachment loss(AL).Concentrations of formic acid,propionic acid and butyric acid in the supernatant of the GCFs were analyzed by high-performance capil-lary electrophoresis(HPCE).The prediction ability of formic acid,propionic acid and butyric acid with the risk of periodontitis and the differences between grade B and grade C periodontitis were analyzed.Results:In this study,32 patients with stage Ⅲ and 5 patients with stage Ⅳ were enrolled,including 9 patients with grade B and 28 patients with grade C.Clinical periodontal variables in the patients with pe-riodontitis were significantly higher than those in the control group(P＜0.001).Formic acid was signifi-cantly lower in periodontitis than that in the control group[5.37(3.39,8.49)mmol/L vs.12.29(8.35,16.57)mmol/L,P＜0.001].Propionic acid and butyric acid in periodontitis were significantly higher than those in the control group:Propionic acid,10.23(4.28,14.90)mmol/L vs.2.71(0.00,4.25)mmol/L,P＜0.001;butyric acid,2.63(0.47,3.81)mmol/L vs.0.00(0.00,0.24)mmol/L,P＜0.001.There was no significant difference in formic acid,propionic acid and butyric acid concentrations between grade B and grade C periodontitis(P＞0.05).Propionic acid and butyric acid in the deep pocket were significantly higher than in the shallow pocket,while the concentration of formic acid decreased with the increase of PD.Propionic acid(OR=1.51,95％CI:1.29-1.75)and butyric acid(OR=3.72,95％CI:1.93-7.17)were risk factors for periodontitis,while formic acid(OR=0.87,95％CI:0.81-0.93)might be a protective factor for periodontitis.Propionic acid(AUC=0.852,95％CI:0.805-0.900),butyric acid(AUC=0.889,95％CI:0.841-0.937),f(formic acid,AUC=0.844,95％CI:0.793-0.895)demonstrated a good predictive capacity for the risk of periodontitis.Conclusion:The concentration of formic acid decrease in the GCF of periodontitis patients,which is a protective factor for periodontitis,its reciprocal have good predictive capacity.However,propionic acid and butyric acid increase,which are risk factors for periodontitis and have good predictive capacity.The concentration of formic acid,propionic acid,and butyric acid vary with probing depth,but there is no significant difference between grade B and grade C periodontitis.

AIM: To explore the promoting effect of 2-APB on skin wound healing in mice and its potential mechanism. METHODS: KM mice were divided into 5 groups: control group, DMSO group, low (50 mg/L), medium (100 mg/L) and high (200 mg/L) concentration 2-APB group. On the back of each mouse's skin use a circular punch about 1 cm on both sides of the midline of the spine to make a skin wound with a diameter of 10 mm and as deep as the fascia. The control group was only wrapped with gauze and no drugs were applied; the DMSO group was applied 1 g DMSO/Vaseline ointment per day; in the 2-APB group, apply 1 g of 2-APB/Vaseline ointment at a corresponding concentration every day. Pictures were taken the next day to observe the healing, and the material was taken on the 21st day, HE staining was used to observe the pathological morphology of the wound and western blot to detect TRPM7, TGF-β, collagen-I and IL-1β expression. RESULTS: Compared with the control group and the DMSO group, different concentrations of 2-APB could significantly promote skin wound healing in mice (P<0.01), but there was no significant difference in wound healing rate between the DMSO group and the control group group. The results of HE staining showed that, compared with the control group group and the DMSO group, 2-APB could increase the collagen content of the wound and the thickness of the dermis (P<0.01), but there was no significant difference between the DMSO group and the control group group. At the same time, 2-APB could also significantly increase the expression of TGF-β and Col-I on the wound, and inhibit the expression of TRPM7 and IL-1β. CONCLUSION: Different concentrations of 2-APB (50, 100 and 200 mg/L) can promote skin wound healing, and its mechanism may be related to the inhibition of TRPM7.

Objective To compare the clinical efficacies of transanal total mesorectal excision(TaTME) and laparoscopic total mesorectal excision (LapTME)for rectal cancer (RC).Methods The case-control matching method and retrospective cohort study were conducted.The clinicopathological data of 100 RC patients who were admitted to the Sixth Affiliated Hospital of Sun Yat-sen University between July 2014 and January 2016 were collected.Of 100 patients,50 undergoing TaTME and 50 undergoing LapTME were respectively allocated into the TaTME and LapTME groups by case-control matching method.Observation indicators:(1) operation situations:operation time,volume of intraoperative blood loss,cases with intraoperative complications and preventive stoma;(2) postoperative recovery:time for diet intake,time for out-of-bed activity,occurrence of complications within 30 days postoperatively and duration of hospital stay;(3) postoperative pathological examinations:postoperative pathological specimen length,number of lymph node harvest,distance from lower boundary of tumor to distant margin and cases with positive circumferential margin;(4) follow-up.Follow-up using outpatient examination and network tracing was performed to detect local tumor recurrence and distant metastasis up to December 2016.Measurement data with normal distribution were represented as x±s and comparison between groups was analyzed using the paired-samples t test.Measurement data with skewed distribution were represented as M (range).Comparisons of count data were analyzed using the chi-square test.Comparisons of measurement data with skewed distribution and ranked data were done by the nonparametric test.Results (1) Operation situations:operation time,volume of intraoperative blood loss,cases with intraoperative complications and preventive stoma were (259±111)minutes,100 mL (range,20-2 000 mL),2,28 in the TaTME group and (220± 80)minutes,50 mL (range,20-1 000 mL),1,33 in the LapTME group,respectively,with no statistically significant difference (t=1.90,Z=-0.30,x2 =0.34,0.01,P＞0.05).(2) Postoperative recovery:time for diet intake and time for out-of-bed activity were (1.6±0.5) days,(2.6±0.6) days in the TaTME group and (2.4±0.5)days,(3.5 ±0.6)days in the LapTME group,respectively,with statistically significant differences (t =8.90,11.30,P＜0.05).Cases with anastomotic fistula,bleeding and stenosis,intestinal obstruction,abdominal abscess and wound infection within 30 days postoperatively were 6,1,1,0,1,0 in the TaTME group and 5,1,2,2,1,2 in the LapTME group,respectively,with no statistically significant difference (x2=0.10,0.00,0.30,2.00,0.00,2.00,P＞0.05).Cases with urinary retention within 30 days postoperatively were 3 and 0 in the TaTME and LapTME groups,respectively,with a statistically significant difference (x2 =3.00,P＜0.05).Two and 2 patients with anastomic fistula underwent reoperation in the TaTME and LapTME groups respectively,and other patients were improved by symptomatic treatment.Duration of hospital stay was 7 days (range,5-36 days)and 8 days (range,6-29 days) in the TaTME and LapTME groups,respectively,with no statistically significant difference (Z =-0.90,P ＞ 0.05).(3) Postoperative pathological examinations:postoperative pathological specimen length,number of lymph node harvest,distance from lower boundary of tumor to distant margin and cases with positive circumferential margin were (11±3)cm,13±5,(1.3±0.7)cm,0 in the TaTME group and (12±3) cm,13±5,(1.3±0.7)cm,1 in the LapTME group,respectively,with no statistically significant difference (t=0.50,0.20,0.10,x2=1.00,P＞0.05).(4) Follow-up:100 patients were followed up for 9-27 months,with an average time of 18 months.During the follow-up,distant metastasis and local tumor recurrence were detected in 2,3 patients of TaTME group and in 2,2 patients of LapTME group,respectively,with no statistically significant difference (x2 =0.00,0.20,P＞0.05).Conclusions TaTME for RC is safe and feasible.Compared with LapTME,TaTME not only achieves identical pathological quality without increasing intra-and postoperative complications,but also benefits postoperative recovery of patients.

OBJECTIVETo explore the learning curve of transanal total mesorectal excision (taTME) for rectal cancer.

METHODSClinical data of 60 rectal cancer patients undergoing taTME from July 2014 to April 2016 were retrospectively analyzed. According to the sequence of operation date, 60 patients were divided into four groups (A, B, C, D) with 15 cases in each group. General information and perioperative, especially the operative indexes were compared among four groups.

RESULTSThere were no significant differences in age, sex, preoperative staging, BMI, tumor size among four groups (all P>0.05). The distance from tumor to anal verge in A group was(6.7±2.5) cm, which was significantly different with B group (4.6±1.2) cm, C group (4.5±1.0) cm and D group (4.0±1.0) cm (P=0.000, P=0.000, P=0.001). Ratio of receiving neoadjuvant therapy was 0, 60.0%(9 cases), 26.7%(4 cases) and 26.7%(4 cases) in A, B, C, D groups respectively with significant difference (P=0.004). Ratio of receiving complete taTME was 73.3%(11/15) in A group, 26.7%(4/15) in B group, 13.3%(2/15) in C group and 26.7%(4/15) in D group, while other patients underwent laparoscopy-assisted procedures. This ratio of A group was significantly higher as compared to B, C, D groups (P=0.003). The operation time was significantly different among four groups [A group (223.0±105.2) minutes, B group (299.0±131.0) minutes, C group(278.0±44.8) minutes, D group (246.0±34.0) min, P=0.035]. Fluctuation of operation time was more common in A and B groups, which became stable in C and D groups. Though intra-operative blood loss was not significantly different among four groups [A group (249.0±559.6) ml, B group (288.0±568.1) ml, C group (87.0±43.3) ml, D group (69.0±64.5) ml, P=0.225], but it presented a decline trend in C and D groups. Number of harvested lymph node from postoperative pathological specimen was 10.9±5.9 in A group, 9.6±2.7 in B group, 15.8±4.8 in C group, and 14.2±5.1 in D group, with significant difference among groups (P=0.008; A group vs. C group, P=0.010; B group vs. C group, P=0.002; B group vs. D group, P=0.021). There were no significant differences in specimen length, postoperative complication rate, distal margin distance and hospital stay.

CONCLUSIONA well-skilled laparoscopic colorectal surgeon, by following the standard surgical procedures, are likely to overcome the learning curve smoothly after performing approximately 30 cases of taTME for rectal cancer.

Abdomen ; Aged ; Anal Canal ; Blood Loss, Surgical ; Digestive System Surgical Procedures ; methods ; Female ; Humans ; Laparoscopy ; Learning Curve ; Length of Stay ; Lymph Nodes ; Male ; Middle Aged ; Neoadjuvant Therapy ; Operative Time ; Postoperative Complications ; Rectal Neoplasms ; surgery ; Retrospective Studies

Objective To investigate the clinical efficacy of unidirectional-loop caudal-medial approach for laparoscopic-assisted radical resection of right colon cancer.Methods The retrospective and descriptive study was performed.The clinical data of 37 patients who underwent laparoscopic-assisted radical resection of right colon cancer through unidirectional-loop caudal-medial approach at the Sixth Mfiliated Hospital of Sun Yat-sen University from January 2015 to March 2016 were collected.Tumor-free principle was followed and unidirectional-loop caudal-medial approach was conducted.Observation indicators included:(1) surgical situations:operation time,volume of intraoperative blood loss,(2) postoperative recovery:time to initial anal exsufflation,time of draining tube removal,postoperative complications,duration of postoperative hospital stay,(3) postoperative pathological examination:number of lymph node dissection,number of positive lymph node,length of specimen,incision margin,tumor pathological staging and type,(4) follow-up.All the patients were followed up using outpatient examination and telephone interview up to June 2016.Measurement data with normal distribution were presented as x ± s and measurement data with skewed distribution were presented as average (range).Results (1) Surgical situations:37 patients received successful operation,without conversion to open surgery and perioperative death.Operation time and volume of intraoperative blood loss in 37 patients were (170 ± 50)minutes and 50 mL (range,20-300 mL).(2) Postoperative recovery:time to initial anal exsufflation,time of draining tube removal and average duration of postoperative hospital stay were (3.5 ± 1.0) days,(4.3 ± 1.1) days and 10 days (range,6-21 days),respectively.Two patients with postoperative wound liquefaction were improved by symptomatic treatment,and the other patients had no complication.(3) Postoperative pathological examination:number of lymph node dissection,number of positive lymph node,number of central lymph node dissection and length of specimen in 37 patients were 22 ±8,0 (range,0-6),6 ±5 and (32 ±9)cm,respectively,with negative incision margins.Postoperative tumor pathological staging showed that stage pT1,pT2,pT3 and pT4a were detected in 0,1,33 and 3 patients,and stage pN0,pN1 and pN2 in 23,12 and 2 patients,respectively.Postoperative tumor pathological type showed that 3,7,23 and 4 patients were respectively diagnosed with mucinous adenocarcinoma,high-differentiated adenocarcinoma,moderate-differentiated adeno-carcinoma and low-differentiated adenocarcinoma.(4) Follow-up:37 patients were followed up for 3-17 months with a median time of 11 months.During the follow-up,1 patient was complicated with anastomotic recurrence and 4 with distant metastases,the other 32 patients had tumor-free survival.Conclusion Unidirectional-loop caudal-medial approach for laparoscopicassisted radical resection of right colon cancer is safe and feasible,with a good short-term outcome,and it should be widely spread.

Objective To investigate the clinical features, effective treatment, survival and prognostic factors of second primary tongue squamous cell carcinoma (SPTSCC) after nasopharyngeal carcinoma (NPC) radiotherapy. Methods The clinical data of 35 cases with SPTSCC after NPC radiotherapy were analyzed retrospectively. Kaplan-Meier method, Log-Rank test and COX proportional hazard mode was performed for statistical analysis. Results 3-year and 5-year overall survival rates were 55 % and 47 %, respectively, lymph node metastasis rate was 5.71 %. Univariate analysis indicated that gender (χ2 = 8.89, P = 0.00), T classification (χ2= 5.58, P= 0.02), clinical stage (χ2 = 8.51, P= 0.04) and treatment methods (χ2 = 29.37, P = 0.00) were important factors of prognosis. Multivariate analysis showed that treatment methods (P = 0.00) and T classification (P = 0.03) were independent prognostic factors. Operative treatment group had better prognosis than the non-operative treatment group, the difference was statistically significant (P <0.05), male patients in the risk of SPTSCC was higher than the female patients, and the incidence of SPTSCC was increased along with extension of the time after NPC radiotherapy. Conclusion The rate of the lymph node metastasis is lower for SPTSCC after NPC radiotherapy and treatment patterns and T stage are independent prognostic factors. Long-term follow-up after NPC radiotherapy is necessary to the early diagnosis of SPTSCC, so that to give surgery or combined therapy with surgery in order to achieve a good effect.