Objective : To investigate the effects of melatonin ( MEL) on the proliferation of hepatic stellate cells (HSCs) induced by platelet - derived growth factor (PDGF⁃BB) and explore its correlation with the regulation of autophagy levels . Methods : The HSC⁃T6 cells were divided into the following groups : control group , model group and MEL (low , medium and high) treatment groups . After 24 hours culture , the cells adhered to the wall and were changed into serum⁃free DMEM medium to synchronize the cells to the G0 stage . After 24 hours culture , all groups were given with PDGF ⁃ BB ( 10 ng/ml) excepted the control group . Besides , melatonin of different concentrations ( 1 nmol/L , 1 μmol/L and 0. 1 mmol/L) were added immediately in three treated groups . After incubated for 48 hours , the effect of MEL on the proliferation of hepatic stellate cells activated by PDGF⁃BB was detected by CCK8 method . The protein expression levels of LC3b and α ⁃SMA in hepatic stellate cells were determined by Western blot . The expression levels of LC3b mRNA and α ⁃SMA mRNA in hepatic stellate cells were determined by qRT⁃PCR . The ultrastructure of HSCs was observed by transmission electron microscopy to understand the autophagy level. Results : Compared with control group , PDGF⁃BB could induce the proliferation of HSCs ( P < 0. 01) . Compared with model group , MEL inhibited the proliferation of HSCs activated by PDGF⁃BB ( P < 0. 01) . Compared with the control group , LC3b and α ⁃SMA protein expressions significantly increased in the model group ( all P < 0. 05) , and LC3b mRNA and α ⁃SMA mRNA expressions significantly increased in the model group (P < 0. 05 , P < 0. 01) . Compared with the model group , MEL could inhibit such effects (LC3b : P < 0. 05 , P < 0. 01 ; α ⁃SMA : P < 0. 01) . Transmission electron microscopy ( TEM) showed that compared with the control group , autopolysosome significantly increased in the model group (P < 0. 05) . Compared with model group , autopolysosome significantly decreased in MEL treatment group (P < 0. 01) . Conclusion The up⁃regulation of autophagy level can promote the proliferation of hepatic stellate cells and the inhibition of hepatic stellate cell proliferation by MEL may be related to the down⁃regulation of autophagy level .

Objective: To investigate the oxidative stress injury of nano zinc oxide nanoparticles(ZnO NPs) on human myocardial cells (AC16) ，and to analyze the mechanism of ZnO NPs from the transcriptome level． Methods: Dynamic light scattering (DLS) was used to characterize and detect ZnO NPs．After AC16 cells were exposed to ZnO NPs at different doses and at different times，the cell survival rate was determined by CCK-8 method．AC16 cells were divided into control group，ZnO NPs (50，100，200 μmol /L) ，after 6 h treatment，the mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) were measured．AC16 cells were divided into control group，50 μmol /L ZnO NPs group and 200 μmol /L ZnO NPs group．After 6 h exposure，total RNA was extracted by TRIzol for transcriptome analysis ，and the differentially expressed genes were enriched by gene body ( GO) ，Kyoto Encyclopedia of Genes and Genomes (KEGG) . Results : The results of DLS showed that the hydrodynamic diameter was ( 192. 2 ± 1. 63 ) nm and the Zeta potential was ( －23. 26 ± 1. 05 ) mV． CCK-8 results showed that the survival rate of AC16 cells decreased with the increase of dose and time of exposure to ZnO NPs. Fluorescence quantification showed that with the increase of ZnO NPs exposure dose，MMP significantly decreased at 100 μmol /L ZnO NPs(P＜0. 05) ，and ROS significantly increased at 50 μmol /L ZnO NPs(P＜0. 05) ．Using the multifunctional microplate reader，it was observed that MMP and ROS were statistically significant at 100 and 50 μmol /L ZnO NPs，respectively，showing a decrease in MMP and an increase in ROS．Transcriptome analysis showed that 1 071 genes were enriched in the 50 μmol /L ZnO NPs group compared with the control group，including 561 up-regulated genes and 510 down-regulated genes．Compared with the control group，7 164 genes were enriched in 200 μmol /L ZnO NPs group，including 4 098 up-regulated genes and 3 066 down-regulated genes．GO and KEGG analysis showed that the differential genes were mainly concentrated in ROS，antioxidant activity，mitochondrial cytochrome C release，apoptosis and other signaling pathways． Conclusion ZnO NPs can decrease the survival rate of AC16 cells and induce mitochondrial damage and oxidative stress，among which ROS-mediated oxi- dative stress and mitochondrial function changes are important toxic mechanisms of ZnO NPs induced AC16 cytotoxicity.

Preoperative treatment of colorectal cancer includes neoadjuvant therapy for initial resectable patients and conversion therapy for initial unresectable patients. In locally advanced rectal cancer, on the basis of neoadjuvant chemoradiotherapy or radiotherapy, increasing the intensity of concurrent chemotherapy, or raising postoperative adjuvant chemotherapy before surgery, or combining with immunotherapy can increase pathological downstaging, contribute to organ preservation, and improve survival of patients. In locally advanced colon cancer, neoadjuvant chemotherapy can improve surgical outcomes. In patients with resectable colorectal liver metastases, neoadjuvant chemotherapy is recommended in patients with unfavorable prognostic factors, but it remains controversial whether it should be combined with targeted therapy. However, in patients with initially unresectable colorectal liver metastases, under the guidance of molecular typing, chemotherapy, especially triple-drug chemotherapy, combined with targeted therapy, is expected to achieve higher objective response rate and convertible rate, thus accepting surgical resection, which improves long-term survival. In addition, for the patients with mismatch repair deficient/micro-satellite instability-high metastatic colorectal cancer, programmed death-1 monoclonal antibody (mAb) and/or cytotoxic T lymphocyte-associated antigen-4 mAb have become the standard first-line treatment option.

Objective: To explore the mechanism of maternal vitamin D deficiency(VDD)-induced fetal intrauterine growth restriction( IUGR) during pregnancy. Methods: Four weeks female CD-1 mice were divided into two groups: control mice( CTRL) and vitamin D deficiency mice( VDD). VDD mice were fed with diet with low concentration of vitamin D and mated with normal male mice. Several pregnant mice were killed on gestational sixteenth day,and the placentas,serum of maternal and fetal mice were collected. Proteins of placental key enzymes of steroid hormone synthesis and signaling pathways of oxidative stress were measured using Western blot. Serum estrogen,progesterone and 25-( OH)-D were measured via radioimmunoassay. The remaining pregnant mice were killed. Placentas and fetus were weighted,fetus height and placental diameter were calculated. The pregnancy outcomes were observed. Results: Diet with low concentration of vitamin D were used and VDD mice model were established after five weeks. Maternal VDD during pregnancy decreased the weight of placenta and fetus,reduced the height of fetus and the diameter of placentas. The expression of placental aromatase cytochrome p450( CYP19) and the level of estrogen were increased in VDD pregnant mice. Besides,maternal VDD during pregnancy increased the expressions of placental heme oxygenase 1( HO-1),NADPH oxidase 4( NOX-4) and 3-nitrotyrosine( 3-NT).VDD promoted placental nuclear factor E2-related factor 2( Nrf-2) nuclear translocation. Conclusion Maternal vitamin D deficiency during pregnancy causes IUGR partially through evoking placental oxidative stress and promoting estrogen synthesis.

Objective:To investigate the risk factors for lymph node metastasis in T1 colorectal cancer and application value of its nomogram prediction model.Methods:The retrospective case-control study was conducted. The clinicopathological data of 914 patients with T1 colorectal cancer who underwent radical resection in the Zhongshan Hospital of Fudan University from June 2008 to December 2019 were collected. There were 528 males and 386 females, aged from 25 to 87 years, with a median age of 63 years. Observation indicators: (1) clinicopathological data of patients with T1 colorectal cancer; (2) follow-up; (3) analysis of influencing factors for lymph node metastasis; (4) development and internal validation of a nomogram predition model. Patients were regularlly followed up once three months within postoperative 2 years and once six months thereafter to detect tumor recurrence and survival. The endpoint of follow-up was at postoperative 5 years. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. The Kaplan-Meier method was used to calculate survival rates and draw survival curves. The Log-rank test was used for survival analysis. Univariate and multivariate analyses were performed using the Logistic regression analysis. Based on results of multivariate analysis, a Logistic regressional nomogram for prediction of lymph node metastasis probability was constructed using R language software. The calibration curve was used to evaluate the consistency between probability predicted by the nomogram model and actual observation probability, which was reprensented by a consistency index. The Bootstrap method was used for evaluation of the model performance to receive the calibration curve. The Hosmer-Lemeshow test was used to calculate the goodness of fit in model. Results:(1) Clinicopathological data of patients with T1 colorectal cancer: 687 of 914 patients underwent direct surgery and 227 underwent remedial operation after endoscopic resection. All the 914 patients were confirmed as pT1NxM0 colorectal cancer by pathological examination. The tumor diameter was (2.3±1.2)cm. The pathological catogaries of 914 patients included 865 cases of adenocarcinoma and 49 cases of mucinous adenocarcinoma. The tumor differentiation degree of 914 patients included 727 cases of high or middle differentiation and 187 cases of low differentiation or undifferentiation. Of the 914 patients, 633 cases had submucosal infiltration depth ≥1 000 μm and 281 cases had submucosal infiltration depth <1 000 μm. There were 110 cases with nerve vessel invasion and 804 without nerve vessel invasion. The number of intraoperative lymph node dissection was 13 (range, 1-48). There were 804 cases in stage N0 of N staging, 98 cases in stage N1 and 12 cases in stage N2. There was no perioperative death. (2) Follow-up: 886 of 914 patients were followed up for 25 months (range, 1-129 months). During the follow-up, 24 patients had tumor recurrence or metastasis. The 5-year cumulative tumor recurrence rate of 914 patients was 4.8% and the median recurrence time was 17.0 months. Liver was the main site of tumor recurrence, accounting for 58.3%(14/24). The 5-year recurrence-free survival rate of 914 patients was 95.2%. The 5-year recurrence-free survival rate was 96.3% of 804 patients without lymph node metastasis, versus 86.6% of 110 patients with lymph node metastasis, showing a significant difference between the two groups ( χ2=6.83, P<0.05). (3) Analysis of influencing factors for lymph node metastasis: results of univariate analysis showed that preoperative carcinoembryonic antigen (CEA), preoperative CA19-9, tumor differentiation degree, submucosal infiltration depth, nerve vessel invasion were related factors for lymph node metastasis in T1 colorectal cancer ( odds ratio=2.56, 3.25, 2.21, 2.68, 3.39, 95% confidence interval as 1.41-4.67, 1.22-8.66, 1.43-3.41, 1.56-4.88, 2.10-5.48, P<0.05). Results of multivariate analysis showed that preoperative CEA ≥5 μg/L, preoperative CA19-9 ≥37 U/mL, poor differentiation or undifferentiation, submucosal infiltration depth ≥1 000 μm and nerve vessel invasion were independent risk factors for lymph node metastasis in T1 colorectal cancer ( odds ratio=2.23, 3.47, 2.01, 2.31, 2.91, 95% confidence interval as 1.02-4.15, 1.08-10.87, 1.03-3.27, 1.40-4.47, 1.64-5.13, P<0.05). (4) Development and internal validation of a nomogram predition model: based on results of multivariate Logistic analysis, a nomogram prediction model for lymph node metastasis in T1 colorectal cancer was developed. The nomogram score was 59 for preoperative CEA >5 μg/L, 100 for preoperative CA19-9 ≥37 U/mL, 48 for poor differentiation or undifferentiation, 67 for submucosal infiltration depth ≥1 000 μm and 92 for nerve vessel invasion, respectively. The total of different scores for different clinicopathological factors corresponded to the probability of lymph node metastasis. The receiver operating characteristic curve was drawed to evaluate the predictive performance of nomogram for lymph node metastasis in T1 colorectal cancer, with the area under curve of 0.70(95% confidence interval as 0.64-0.75, P<0.05). The Bootstrap internal validation of predictive performance in the nomogram predition model showed a consistency index of 0.70 (95% confidence interval as 0.65-0.75). The calibration chart showed a good consistency between the probability predicted by the nomogram model and actual probability of lymph node metastasis. The Hosmer-Lemeshow test showed a good fitting effect in model ( χ2=1.61, P>0.05). Conclusions:Preoperative CEA ≥5 μg/L, preoperative CA19-9 ≥37 U/mL, poor differentiation or undifferentiation, submucosal infiltration depth ≥ 1 000 μm and nerve vessel invasion are independent risk factors for lymph node metastasis in T1 colorectal cancer. The constructed nomogram model can help predict the probability of lymph node metastasis in T1 colorectal cancer.

Rheumatoid arthritis (RA) is an autoimmune disease and is mainly characterized by abnormal proliferation of fibroblast-like synoviocytes (FLS). The up-regulated cellular membrane expression of G protein coupled receptor kinase 2 (GRK2) of FLS plays a critical role in RA progression, the increase of GRK2 translocation activity promotes dysfunctional prostaglandin E4 receptor (EP4) signaling and FLS abnormal proliferation. Recently, although our group found that paeoniflorin-6'-

The Corona Virus Disease 2019 (COVID-19) since December, 2019 has a wide range of infection due to the strong infectious characteristics. Both medical staff and patients are at increased risk of infection. It is an urgent clinical problem for specialist doctors to work with diagnosis and treatment of cancer patients during the epidemic situation. Based on the colorectal cancer diagnosis and treatment guidelines (2019 CSCO guideline), combined with their own experience, the authors propose the overall management strategies for colorectal cancer patients. This strategies cover the key diagnosis and treatment of colorectal cancer, and provide targeted clinical practice. These work will be helpful for colorectal cancer specialists to carry out the diagnosis and treatment of colorectal cancer effectively under the epidemic of COVID-19.

Objective:To investigate the risk factors associated with anastomotic leakage after robotic surgery in mid-low rectal cancer.Methods:A retrospective case-control study method was conducted. Inclusion criteria: (1) 18 to 80 years old; (2) pathologically confirmed rectal cancer; (3) distance <10 cm from tumor to anal margin; (4) robotic anterior rectal resection. Patients with previous history of colorectal cancer surgery, distant metastases or other malignant tumors, undergoing emergency surgery, with severe abdominal adhesions or those receiving combined organ resection were excluded. Based on the above criteria, 636 patients undergoing robotic radical sphincter-preserving surgery for mid-low rectal cancer in Zhongshan Hospital from January 2015 to December 2018 were included in this study, including 398 males (62.6%) and 238 females (37.4%) with a mean age of (61.9±11.3) years. Sixty-eight cases (10.7%) received neoadjuvant chemoradiotherapy. Amony the 636 included patients, 123(19.3%) underwent natural orifice specimen extraction surgery (NOSES) and 15 (2.3%) underwent preventive stoma. According to the cirteria developed by the International Rectal Cancer Research Group in 2010, the anastomotic leakage was classified as grade A (no requirement of intervention), B (requirement of intervention), and C (requirement of operation). Logistic regression was used to analyze the relationship between anastomotic leakage and clinicopathological factors. Factors in univariate analysis with P<0.05 were included in the multivariate analysis. Results:Anastomotic leakage occurred in 38 cases (6.0%). The grading of anastomotic leakage was grade A in 13 cases (2.0%), grade B in 19 cases (3.0%), and grade C in 6 cases (0.9%). The 3-year disease-free survival rate of patients with anastomotic leakage and without anastomotic leakage was 83.5% and 83.6% respectively ( P=0.862); the 3-year overall survival rate of the two group was 85.1% and 87.5% respectively ( P=0.296). The results of univariate logistic regression analysis showed that male ( P=0.011), longer operation time ( P=0.042), distance ≤5 cm from tumor to anal margin ( P=0.012), more intraoperative blood loss ( P=0.048) were associated with anastomotic leakage (all P<0.05). NOSES was not associated with anastomotic leakage ( P=0.704). Multivariate analysis confirmed that male (OR=3.03, 95%CI: 1.37 to 7.14, P=0.010), operation time ≥180 minutes (OR=2.04, 95%CI: 1.03 to 3.99, P=0.040), distance ≤5 cm from tumor to anal margin (OR=2.56, 95%CI:1.28 to 5.26, P=0.008) were independent risk factors for anastomotic leakage. Conclusion:Male, short distance from tumor to anal margin, and long operation time are independent risk factors for anastomotic leakage in patients undergoing robotic mid-low rectal cancer radical surgeries. These patients need to be cautiously treated during surgery.

Objective:To investigate the short-term outcomes of laparoscopic simultaneous resection of primary colorectal cancer and liver metastases in patients with resectable synchronous colorectal liver metastases (SCRLM).Methods:A descriptive case series study was performed. Clinicopathological data of patients with SCRLM who underwent laparoscopic simultaneous resection of colorectal cancer and liver metastases in Zhongshan Hospital between December 2015 and September 2018 were retrieved from a prospective colorectal cancer database. Perioperative presentations and short-term outcomes were analyzed.Results:A total of 53 patients were enrolled with average age of(61.7±11.3) years. Among them, 32 were male (60.4%) and 21 were female (39.6%). Twenty-five patients (47.2%) were American Society of Anesthesiologists (ASA) grade I and 28 (52.8%) were grade II. All the patients completed laparoscopic simultaneous resection without conversion. The average operation time was (320.2±114.5) min. The estimated blood loss was 150.0 (45.0-2000.0) ml, and only 2 cases (3.8%) received intraoperative transfusion. Postoperative pathologic results revealed that the average primary tumor size was (5.4±1.9) cm; 4 cases (7.5%) were T1-2 stage and 48 cases (90.6%) were T3-4 stage; 40 patients (75.5%) had lymph node metastasis; 19 (35.8%) had vascular involvement; 24 (45.3%) had neural invasion. The median number of liver metastases was 1.0 (1-8), and the average size of largest liver metastases was (3.0±1.9) cm. The median margin of liver metastases was 1.0 (0.1-3.5) cm, and only 1 case was R1 resection. The average time to the first postoperative flatus was (67.9±28.9) h, and the average time to the liquid diet was (107.0±33.8) h. The average postoperative indwelling catheterization time was (85.6±56.4) h. The average postoperative hospital stay was (9.2±4.4) d, and the average cost was (82±26) thousand RMB. No death within postoperative 30-day was found. The morbidity of postoperative complication was 32.1% (17/53) and 3 patients developed grade III to IV complications which were improved by conservative treatment. The median follow-up period was 23.2 months. During follow-up, 19 patients (35.8%) developed recurrence or metastasis, and 4 (7.5%) died. The 1- and 2-year disease-free survival (DFS) rates were 68% and 47% respectively, and the 1- and 2-year overall survival rates were 95% and 86% respectively.Conclusions:Laparoscopic simultaneous resection of primary colorectal cancer and liver metastases is safe and feasible in selected patients with SCRLM. Postoperative intestinal function recovery is enhanced, and morbidity and oncological outcomes are acceptable.

Objective:To investigate the risk factors associated with anastomotic leakage after robotic surgery in mid-low rectal cancer.Methods:A retrospective case-control study method was conducted. Inclusion criteria: (1) 18 to 80 years old; (2) pathologically confirmed rectal cancer; (3) distance <10 cm from tumor to anal margin; (4) robotic anterior rectal resection. Patients with previous history of colorectal cancer surgery, distant metastases or other malignant tumors, undergoing emergency surgery, with severe abdominal adhesions or those receiving combined organ resection were excluded. Based on the above criteria, 636 patients undergoing robotic radical sphincter-preserving surgery for mid-low rectal cancer in Zhongshan Hospital from January 2015 to December 2018 were included in this study, including 398 males (62.6%) and 238 females (37.4%) with a mean age of (61.9±11.3) years. Sixty-eight cases (10.7%) received neoadjuvant chemoradiotherapy. Amony the 636 included patients, 123(19.3%) underwent natural orifice specimen extraction surgery (NOSES) and 15 (2.3%) underwent preventive stoma. According to the cirteria developed by the International Rectal Cancer Research Group in 2010, the anastomotic leakage was classified as grade A (no requirement of intervention), B (requirement of intervention), and C (requirement of operation). Logistic regression was used to analyze the relationship between anastomotic leakage and clinicopathological factors. Factors in univariate analysis with P<0.05 were included in the multivariate analysis. Results:Anastomotic leakage occurred in 38 cases (6.0%). The grading of anastomotic leakage was grade A in 13 cases (2.0%), grade B in 19 cases (3.0%), and grade C in 6 cases (0.9%). The 3-year disease-free survival rate of patients with anastomotic leakage and without anastomotic leakage was 83.5% and 83.6% respectively ( P=0.862); the 3-year overall survival rate of the two group was 85.1% and 87.5% respectively ( P=0.296). The results of univariate logistic regression analysis showed that male ( P=0.011), longer operation time ( P=0.042), distance ≤5 cm from tumor to anal margin ( P=0.012), more intraoperative blood loss ( P=0.048) were associated with anastomotic leakage (all P<0.05). NOSES was not associated with anastomotic leakage ( P=0.704). Multivariate analysis confirmed that male (OR=3.03, 95%CI: 1.37 to 7.14, P=0.010), operation time ≥180 minutes (OR=2.04, 95%CI: 1.03 to 3.99, P=0.040), distance ≤5 cm from tumor to anal margin (OR=2.56, 95%CI:1.28 to 5.26, P=0.008) were independent risk factors for anastomotic leakage. Conclusion:Male, short distance from tumor to anal margin, and long operation time are independent risk factors for anastomotic leakage in patients undergoing robotic mid-low rectal cancer radical surgeries. These patients need to be cautiously treated during surgery.