AIM: To set up a method of inducing mouse embryonic stem cells (mESC) to differentiate into cardiomyocyte after treatment with 5-azacytidine. METHODS: Cytotoxicity of 5-azacytidine was measured by MTT assay. Treatment of mESC with conditioned culture mediums, which were composed of 5-azacytidine alone or combined with retinoic acid, induced the cell differentiation to cardiomyocytes. The cells induced were identified by detecting the expression of cardiac proteins (myosin, desmin, ?-actin and ?-actinin). Gene MLC-2v, a specific gene of ventricular-like cardiomyocyte, was also detected by RT-PCR. RESULTS: The non-cytotoxic dose of 5-azacytidine was 8 ?mol/L, which was able to induce mESC to differentiate into cardiac syncytiums. Cells induced expressed many cardiac proteins and MLC-2v mRNA. However, combined with retinoic acid inhibited mESC differentiation into cardiomyocyte. CONCLUSION: 5-azacytidine is able to promote mESC differentiation into cardiomyocytes. A method of inducing mESC to differentiate into cardiomyocytes in vitro has been established.

Objective To investigate the effect of oil with medium-long-chain triacylglycerols on blood lip- id and lipoproteins in hyperlipemic patients. Methods Totally, 112 patients with hypertriglyceridemia were en- rolled and randomly divided into MLCT group (consumed oil with medium-long-chain fatty acids) and LCT group (consumed oil with long-chain fatty acids) (both 25-30 g/d for 8 weeks). Patients in both two groups were also instructed to take exercises. Height and weight were measured at baseline and 8 weeks later. Blood glucose, serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterols (TC), triglyc- erides (TG), high density lipoprotein cholesterol, low density lipoprotein cholesterol, apolipoprotein Al (ApoAl), ApoB, ApoA II , ApoC2, ApoC3, and ApoE were measured and compared. Results At the end of study, 101 subjects were included. There were 50 subjects left in LCT group and 51 subjects left in MLCT group, respectively. There was no significant difference in weight, ALT, AST, TC, and TG at baseline between two groups (P>0.05). Eight weeks later, weight, serum TG, ApoC2, and ApoC3 were significantly lower and ApoAl level was significantly higher than those at baseline in MLCT group (P < 0.05). At the end of study, the decreases in body weight and blood biochemical variables including TG, ApoB, ApoA II , ApoC2, ApoC3 were significantly much greater in MLCT groups than those in LCT group (P < 0. 05). Conclusion When the diet is reasonably controlled, oil of medium-long-chain triacylglycerols may reduce the concentration of TG and improve the levels of apolipoproteins.

Objective To explore the feasibility,the safety,D2 lymph node dissection and clinical outcomes of laparoscope-assisted radical gastrectomy lot advanced gastric cancer.Methods The clinica data of 47 cases with advanced gastric treated with laparoscope-assisted radical gastrectomy were analyzed retrospectively.Results Laparoscope-assisted radical gastrectomy,proximal gastrectomy in 25 cases,distalgastrectomy in 11 cases,total gastrectomy in 10 cases;46 were performed Laparoscope-assisted surgery radical gastrectomy successfuly,and the other one was converted to abdominal opening.The average operative time for proximal gastrectomy,distal gastrectomy,and total gastrectomy was(220±55)min,(284±37)min,and (330±50)min,respectivel.The average blood loss in proximal gastrectomy,distal gastrectomy,and total gastrectomy was(150±87)ml,(120±70)ml,and(330±50)ml respectively.The average time of proximal gastrectomy for gastrointestinal function recovery,for the patients to stand up,and for the postoperative hospital stay was(5.1±0.5)d,(3.2±0.8)d and(9.0±1.5)d.The average time of distal gastrectomy for gastrointestinal function recovery,for the patients to stand up,and for the postoperative hospital stay was (4.0±0.8)d,(3.2±1.5)d and(9.0±2.0)d.The average time of total gastrectomy for gastrointestinal function recovery,for the patients to stand up,and for the postoperative hospital stay was(4.1±0.8)d,(3.2±0.8)d and(9.5±2.0)d.The mean total number of retrieved lymph nodes was(21.95±9.88),and the lengths of proximal and distal margins to the tumor were(6.41±2.13)cm and(6.22±1.98)cm respectively.No postoperative deaths or anastomtic fistulas were found.Its short-term outcomes were satisfactory.Conclusion Laparoscope-assisted radical gastrectomy with D2 lymphadenectomy is safe,feasible,which achieves adequate cancer clearance,but the long-term outcome is needed to be observed.

Objective To compare the effects of oil with medium-long-chain triacylglycerols (MLCT) on lipid metabolism in hypertriglyceridemic patients with different sex. Method One hundred and one subjects with hypertriglyceridemia were enrolled and divided into two groups. One group consumed oil with MLCT (50 subjects) and another group with long-chain fatty acid triglyceride LCT (51 subjects) for 8 w. No sex difference was noted between two groups. All subjects were informed to consume 25-30g oils daily with fixed energy and perform exercise informed. Anthropometric measurements of body weight,waist circumference (WC) body fat weight, total visceral and subcutaneous fat area in abdomen and blood biochemical variables of aspartate amino- transferase (AST), alanine aminotransferase (ALT), glucose, total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein A1 (ApoA1), ApoB, ApoAⅡ, ApoC2, ApoC3 and ApoE were measured at initial and final time of the experiment. Results The decreases in body weight, BMI, WC, body fat weight, total and subcutaneous fat area, and blood TG, LDL-C, ApoAⅡ, ApoB, ApoC2, ApoC3, ApoE in male subjects consuming MLCT were much greater than those consuming LCT after 8 w. However, there were no significant differences in these parameters in female subjects between two groups. Conclusion Consumption of oil with MLCT reduces body weight, body fat and blood TG and improves apolipoprotein metabolism in male hypertriacylglycerolemic subjects, but not in female.

Objective:To explore the risk factors of hemothorax after rib fracture and evaluate its predictive value for hemothorax.Methods:A retrospective case control study was made on the data of 449 patients with rib fracture admitted to Jiangsu Shengze Hospital affiliated to Nanjing Medical University from January 2018 to November 2019. There were 308 males and 141 females，with the age range of 19-97 years［（57.4±14.0）years］. The hemothorax was defined as pleural effusion on chest CT or X examination on admission or within one week after admission. There were 330 patients in hemothorax group and 119 patients in non-hemothorax group. Indices were compared between the two groups，including gender，age，occupation，weight，height，underlying diseases［diabetes，chronic obstructive pulmonary disease（COPD），hypertension，hyperlipidemia］，causes of injury，imaging findings［number of rib fracture，flail chest，bilateral rib fractures，locations of rib fracture and intramural injuries（pneumothorax，pulmonary contusion，mediastinal emphysema and myocardial contusion）］，thoracic cavity drainage，injury to admission time，vital signs（blood pressure and heart rate），routine blood［white blood cell，hemoglobin（Hb），platelet，hematocrit（Hct）］，blood type，urine routine（urinary occult blood，urinary protein，urinary ketone body），biochemical examination［total cholesterol（TCHO），triglyceride（TG），high density lipoprotein（HDL-C），low-density lipoprotein（LDL-C），albumin（ALB），total bilirubin（TBIL），glutamic oxalacetic transaminase（AST），alanine transaminase（ALT），urea nitrogen（BUN），creatinine（CRE），glycosylated hemoglobin（HbA1C）］，coagulation tests［prothrombin time（PT），fibrinogen（FIB），plasma D-dimer（D-D），thrombin time（TT）］after admission，trauma score［chest wall injury score（CIS），injury severity score（ISS），new injury severity score（NISS）］and length of hospital stay. The univariate analysis was used to observe the correlation between each factor and hemothorax after rib fracture and to screen the significant correlation factors，followed by multivariate logistic regression analysis to further identify the independent risk factors. The receiver operating characteristic（ROC）curve was used to analyze the predictive value of continuous variables in independent risk factors and to calcuate the optimal threshold.Results:The two groups showed no significant differences in gender，occupation，weight，height，diabetes，COPD，hyperlipidemia，injury to admission time，blood pressure，heart rate，platelet，urine protein，urine ketone body，TCHO，HDL-C，TBIL，ALT，CRE，HbA1C or PT（ P>0.05）. The hemothorax group showed significantly decreased Hb，Hct，TG，LDL-C and TT and significantly increased age，number of rib fracture，white blood cell count，AST，FIB，D-D，trauma score（CIS，ISS，NISS）and length of hospital stay when compared to non-hemothorax group（ P<0.05）. There were significant differences in hypertension，causes of injury，flail chest，bilateral rib fractures and locations of rib fracture and urinary occult blood between the two groups（ P<0.05）. The univariate analysis showed that age，hypertension，number of rib fractures，flail chest，bilateral rib fractures，locations of rib fracture（upper chest anterolateral segment，middle chest anterolateral segment，middle chest posterolateral segment，middle chest proximal spinal segment，lower chest posterolateral segment，lower chest proximal spinal segment），pneumothorax，pulmonary contusion，myocardial contusion，thoracic cavity drainage，white blood cell count，urinary occult blood，BUN，FIB，trauma score（CIS，ISS，NISS）and length of hospital stay were significantly associated with hemothorax（ P<0.05）. The multivariate Logistic regression analysis showed that locations of rib fracture（including middle chest posterolateral segment，middle chest proximal spinal segment，lower chest posterolateral segment and lower chest proximal spinal segment），pulmonary contusion，thoracic cavity drainage，BUN and trauma score（CIS，ISS，NISS）were significantly associated with hemothorax after rib fracture（ P<0.05）. The ROC curve analysis of continous variables in independent risk factors showed BUN area under the curve（AUC）of 0.587（95% CI 0.529-0.645），CIS AUC of 0.824（95% CI 0.779-0.870），ISS AUC of 0.789（95% CI 0.739-0.840）and NISS AUC of 0.876（95% CI 0.835-0.917）（ P<0.05），and the optimal thresholds for the above variables were 5.0 mmol/L，2.5 points，15 points and 21.5 points，respectively. Conclusion:Locations of rib fracture（including the middle chest posterolateral segment，middle chest proximal spinal segment，lower chest posterolateral segment，lower chest proximal spinal segment），pulmonary contusion，thoracic cavity drainage，BUN，trauma score（CIS，ISS，NISS）are independent risk factors for hemothorax after rib fracture. BUN>5.0 mmol/L and trauma score（CIS>2.5 points，ISS>15 points，NISS>21.5 points）have significant values in predicting hemothorax.

Objective:To compare the efficacy and prognosis between fluorescent laparoscopic and conventional laparoscopic radical resection of colorectal cancer.Methods:A total of 114 colorectal cancer patients in Bishan District People's Hospital of Chongqing from June 2019 to January 2021 were selected as the research objects, and they were divided into two groups according to the diagnosis and treatment time. Forty-four patients receiving treatment from June 2019 to February 2020 were enrolled as group A, and 70 patients receiving treatment from March 2020 to January 2021 were set as group B. Patients in group A received fluorescent laparoscopic radical resection of colorectal cancer, and patients in group B received conventional laparoscopic radical resection of colorectal cancer. The operation time, intraoperative blood loss, metastatic lymph node clearance rate, clinical efficacy (the overall response rate was calculated as complete remission + partial remission), postoperative complication rate and recurrence rate within 1 year were compared between the two groups.Results:The clearance rate of metastatic lymph nodes was 86.95% (40/46) in group A, which was higher than 72.22% (52/72) in group B, and the difference was statistically significant ( χ2 = 3.55, P = 0.049). There were no statistical differences in operation time and intraoperative blood loss between the two groups (all P > 0.05). The overall response rate was 86.36% (38/44) in group A, which was higher than 74.28% (52/70) in group B, and the difference was statistically significant ( χ2 = 5.03, P = 0.024). The incidence of complications in group A was 9.09% (4/44), which was not significantly different from that in group B (11.42%,8/70) ( χ2 = 0.16, P = 0.692). The recurrence rate within 1 year of group A was 11.36% (5/44), which was lower than that of group B (24.42%, 15/70), but the difference was not statistically significant ( χ2 = 1.89, P = 0.169). Conclusions:Compared with conventional laparoscopy, fluorescent laparoscopic radical resection of colorectal cancer has better clinical efficacy, higher clearance rate of metastatic lymph nodes, and does not increase the incidence of postoperative metastasis and complications.

We aimed to obtain the recombinant aminopeptidase encoded by Listeria monocytogenes (L. monocytogenes) gene lmo1711, and characterized the enzyme. First, the amino acid sequences of Lmo1711 from L. monocytogenes EGD-e and its homologues in other microbial species were aligned and the putative active sites were analyzed. The putative model of Lmo1711 was constructed through the SWISS-MODEL Workspace. Then, the plasmid pET30a-Lmo1711 was constructed and transformed into E. coli for expression of the recombinant Lmo1711. The his-tagged soluble protein was purified using the nickel-chelated affinity column chromatography. With the amino acid-p-nitroaniline as the substrate, Lmo1711 hydrolyzed the substrate to free p-nitroaniline monomers, whose absorbance measured at 405 nm reflected the aminopeptidase activity. The specificity of Lmo1711 to substrates was then examined by changing various substrates, and the effect of metal ions on the catalytic efficiency of this enzyme was further determined. Based on the bioinformatics data, Lmo1711 is a member of the M29 family aminopeptidases, containing a highly conserved catalytic motif (Glu-Glu-His-Tyr-His-Asp) with typical structure arrangements of the peptidase family. The recombinant Lmo1711 with a size of about 49.3 kDa exhibited aminopeptidase activity and had a selectivity to the substrates, with the highest degree of affinity for leucine-p-nitroaniline. Interestingly, the enzymatic activity of Lmo1711 can be activated by Cd²⁺, Zn²⁺, and is strongly stimulated by Co²⁺. We here, for the first time demonstrate that L. monocytogenes lmo1711 encodes a cobalt-activated aminopeptidase of M29 family.