[摘 要] 目的：探究β-半乳糖苷α2,6-唾液酸转移酶1（ST6GAL1）和双特异性磷酸酶26（DUSP26）在结直肠癌（CRC）中的表达及其对巨噬细胞极化和肿瘤进展的影响。方法：选取CRC患者126例，采用免疫组织化学染色法检测肿瘤组织和癌旁组织中ST6GAL1、DUSP26蛋白表达。采用Western blotting、免疫共沉淀、外泌体鉴定、流式细胞术、ELISA、免疫荧光等方法，分析ST6GAL1、DUSP26及其相关信号通路对CRC细胞外泌体介导的M2型巨噬细胞极化、线粒体功能和内质网应激的影响。CRISPR/Cas9和慢病毒系统构建ST6GAL1敲除（ST6GAL1-KO）和DUSP26过表达（DUSP26-OE）细胞模型，并采用小鼠肿瘤模型评估ST6GAL1-KO + DUSP26-OE联合干预对肿瘤生长、细胞增殖及凋亡的影响。结果：CRC患者肿瘤组织中ST6GAL1蛋白阳性率显著高于癌旁正常组织，而DUSP26蛋白阳性率显著低于癌旁正常组织（均P < 0.001）；CRC细胞中ST6GAL1蛋白表达显著高于正常结肠上皮细胞，而DUSP26蛋白表达及p-JNK/JNK、p-c-JUN/c-JUN水平均显著低于正常结肠上皮细胞（均P < 0.05）。CRC细胞来源的外泌体显著促进M2巨噬细胞极化，并上调下游蛋白p-Syk、p-PI3K、p-AKT、线粒体功能障碍蛋白DRP1和内质网应激蛋白CHOP、GRP78蛋白表达（均P < 0.01）。DUSP26-OE组或ST6GAL1-KO组CRC细胞来源的外泌体可抑制上述效应（P < 0.05或P < 0.01）。动物实验显示，ST6GAL1-KO组和DUSP26-OE组肿瘤体积均明显小于Vector组，联合干预组抑瘤效果最为显著（均P < 0.001）；联合干预组细胞增殖标志物PCNA阳性率降低，TUNEL阳性细胞显著增加（P < 0.05或P < 0.001）。ST6GAL1-KO和DUSP26-OE可显著降低小鼠肿瘤组织GRP78和DRP1阳性比例及小鼠体内M2巨噬细胞比例。结论：CRC中ST6GAL1高表达可能通过影响外泌体唾液酸化水平，参与M2型巨噬细胞极化及应激相关蛋白表达调控，DUSP26低表达可能削弱JNK相关通路对ST6GAL1表达的负向调控；两者联合干预对移植瘤生长的抑制作用更明显。

Objective:To report the use of right internal mammary artery (RIMA) in coronary artery bypass grafting (CABG) in our center, summarize the purpose and configuration of RIMA graft in CABG.Methods:All clinical data of coronary artery bypass grafting patients in our center performed in the past 6 years were collected and analyzed retrospectively. Those patients were divided into RIMA group and non-RIMA group according to the use of RIMA. Propensity score matching had been performed before these data was compared. Surgical technique of use of RIMA was summarized.Results:1 537 CABG had been performed from January 1st, 2016 to October 31st, 2021 in our center. Of which, 128 cases were allocated to RIMA group. After propensity score matching having been performed, there was no difference in baseline data between the RIMA group and the non-RIMA group (128 cases), and the RIMA group had more grafts and arterial grafts than the non-RIMA group. The postoperative drainage volume in the RIMA group was more than that of the non-RIMA group. However, there was no statistical significance in difference of transfusion between two groups. Also, there was no difference in postoperative mechanical ventilation time, ICU time and length of stay postoperatively. The postoperative complications were similar between two groups. Postoperative patency rate of the RIMA graft was as high as 95.2%. The target vessels of RIMA included left anterior descending branch (45 cases), diagonal branch (19 cases), intermediate branch (12 cases). obtuse marginal or circumflex branch (16 cases), posterior descending branch (5 cases) and right coronary trunk (18 cases). 41 RIMA used as free grafts, 87 used as in situ grafts, of which 19 RIMA need lengthened by other graft vessels.Conclusion:The patency rate of RIMA graft is high and the application of RIMA do not increase the surgical risk significantly. However, there are versatile contour of RIMA grafts. It can be used as artery graft in selected patients.

Objective:To discuss the indication, risk, postoperative complications and corresponding treatment strategies of minimally invasive coronary surgery.Methods:Reviewed 505 cases of minimally invasive single coronary artery bypass grafting completed in our center from November 2005 to March 2019, including 381 males(75.4%) and 124 females(24.6%). 479 cases were via left anterior small thoracotomy(MIDCAB), 21 cases were assisted by the Davinci system(RA-MIDCAB), and 5 cases were via distal mini-sternotomy(DIMS-CABG). Recorded and analyzed the patient's basic condition, surgical data, postoperative complications and treatment, and postoperative follow-up results.Results:7 cases of MIDCAB were converted into median sternotomy due to LIMA injury or pleural adhesion. Another 7 LIMA injury cases used RIMA or SVG anastomosed with LIMA to complete the revascularization. 6 cases underwent reoperation due to bleeding. 4 cases of postoperative myocardial infarction, 1 of which dead. 2 cases of stroke. 81 patients followed up with coronary CT or arteriography. And there were 8 occlusive grafts(9.88%). 405 patients completed telephone follow-up, all patients survived, good quality of life, no heart or cerebrovascular events.Conclusion:Minimally invasive single coronary artery bypass grafting is a safe and effective operation. Preoperative detailed assessment, careful operation during surgery, adequate contingency planning and reasonable response measures can further reduce the occurrence of complications.

In recent years, adaptive laboratory evolution (ALE) has emerged as a powerful tool for basic research in microbiology (e.g., molecular mechanisms of microbial evolution) and efforts on evolutionary engineering of microbial strains (e.g., accelerated evolution of industrial strains by bringing beneficial mutations). The ongoing rapid development of next-generation sequencing platforms has provided novel insights into growth kinetics and metabolism of microbes, and thus led to great advances of this technique. In this review, we summarize recent advances in the applications of long-term and short-term ALE techniques mainly for microbial strain engineering, and different modes of ALE are also introduced. Furthermore, we discuss the current limitations of ALE and potential solutions. We believe that the information reviewed here will make a significant contribution to further advancement of ALE.
High-Throughput Nucleotide Sequencing ; Laboratories ; Mutation

Objective:To investigate the safety and efficacy of simultaneous carotid endarterectomy (CEA) and coronary artery bypass grafting (CABG) for patients with concomitant severe carotid and coronary artery disease.Methods:The clinical data of 19 patients with concomitant severe carotid artery stenosis and coronary artery disease undergoing simultaneous CEA and CABG at Peking University People′s Hospital from Jan 2011 to Dec 2019 were retrospectively analyzed.Results:The mean ages was 69 years old.The operation adopted the strategy of CEA first and then CABG. Conventional CEA with carotid arterial shunting was performed. The primary composite end points were perioperative cardiovascular and neurological adverse event rates, as well as the late follow-up outcomes. The technical success rate was 100%. There were no adverse cardiovascular events during the perioperative period. Ischemic stroke occurred in 2 patients. No early death was observed. Seventeen cases were successfully followed up for 1-103 months. One patient developed cerebral infarction after 8 months, one developed acute myocardial infarction 43 months after surgery. No cases suffered from carotid artery restenosis. The 5-year overall survival rate was 91%.Conclusions:Simultaneous CEA and CABG treatment for patients with concomitant carotid and coronary artery disease is safe with few perioperative cardiovascular events and no deaths.

Objective:To analyze and compare difference of ultrasonic blood flow of left internal mammary artery(LIMA) and right internal mammary artery(RIMA)as graft to left anterior descending artery(LAD) in patients undergoing off-pump coronary artery bypass grafting(OPCABG).Methods:From October 2017 to October 2019, a total of 363 patients who underwent OPCAB including 329 patients in LIMA-LAD group and 34 patients in RIMA-LAD group, were included in this study. Transthoracic ultrasound examination on IMA was performed before OPCABG. The blood flow , the value of PI(pulsation index) and diastolic flow(DF) of LIMA and RIMA to left anterior descending were measured and recorded by intraoperative TTFM. Patients underwent coronary artery CT examinations at 1 week after OPCABG to discover the patency of grafts.Results:The pre-operative diameter of RIMA was larger and the blood flow and pulsation index of RIMA was better than that of LIMA before OPCABG( P<0.001). But there was no significant difference in the flow, pulsation index and DF value of graft between the two groups after IMA were anastomosed to LAD( P>0.05). In situ skeletonized graft of RIMA did not show the advantage of increased blood flow. Compared with the preoperative parameters of flow of internal mammary artery, both the left and right grafts of IMA were significantly better in blood flow and pulsation index( P<0.001). Considering the remarkable difference in age between the two groups, Flow, PI and DF were compared again after propensity score matching, and there was still no statistical difference between the two groups( P>0.05). A total of 313 patients in LIMA-LAD group completed coronary CTA examination one week after surgery, among which 4 grafts had different degrees of problems. A total of 34 patients in RIMA-LAD group completed coronary CTA examination, one case indicated that the CTA imaging in distal segment was not clear, and the other results had no obvious abnormality. There was no significant difference in postoperative rate of coronary CTA completion( P=0.381) and rate of graft failure( P=0.405) between the two groups. Conclusion:Compared with the preoperative parameters of blood flow of internal mammary artery, both the left and right IMA grafts are significantly better in flow and pulsation index. The pre-operative diameter of RIMA is larger and the blood flow and pulsation index of RIMA is better than that of LIMA before OPCABG. But there is no significant difference in the flow, pulsation index and DF value of graft between the two groups after IMA are anastomosed to LAD. In situ skeletonized graft of RIMA do not show the advantage of increased blood flow.

Objective:To observate the changing trend of flow of in-situ bilateral internal mammary artery as grafts during perioperative period in patients undergoing coronary artery bypass grafting(CABG), and to compare the parameters of flow between LIMA and RIMA.Methods:A total of 57 patients , 50 males and 7 femails, mean aged(57.8±10.1) years, underwent bilateral IMA CABG in our hospital from August 2016 to January 2019. Transthoracic ultrasound of IMA examination was performed before CABG. The blood flow, the PI(pulsation index) and other parameters were measured and recorded by intraoperative TTFM.Results:There was no significant difference of the average diameter and PI between LIMA and RIMA, but the preoperative flow of RIMA is higher( P=0.026) in our study. There was no significant difference of the average flow and PI between LIMA and RIMA recorded by TTFM( P>0.05), but the higher diastolic flow(DF) in LIMA grafts( P=0.022) compared with RIMA grafts may be associated with the different target sites( P<0.05). Compared with the preoperative flow and PI of IMA, the intraoperative flow and PI of both LIMA and RIMA grafts were better( P<0.001). Conclusion:Compared with the preoperative flow of IMA, both flow of LIMA and RIMA are better. In spite of the targets exist difference , there are no significant difference of the average flow and PI between LIMA graft and RIMA graft recorded by TTFM.

Objective:To investigate the early clinical outcomes and experience of minimally invasive coronary surgery-bilateral internal mammary artery(MICS-BIMA) through the left anterior small incision.Methods:132 consecutive patients with 2-vessel disease receiving off-pump coronary artery bypass surgery from October 2015 to June 2019 were enrolled in this study, 20 of which performed MICS-BIMA while 112 cases the routine OPCAB through median sternal incision. Propensity Score Match was used to balance the two groups with the ratio of 1∶3, 60 cases selected as the control group from OPCAB in consequence. After the match, two groups were compared in terms of the baseline and the perioperative data. The conduit patency was evaluated in MICS-BIMA group.Results:No statistically significant difference was shown in the baseline between the two groups( P>0.05). The operation time in MICS-BIMA group was longer than that in OPCAB group[(256.0±57.7)min vs.(227.8±46.8) min, t=2.20, P=0.03)], and the postopende hospital stag nas colso short(nedim 7 days vs. medion 10 ays, P=0.002), the differences were statistically signific cint.. The perioperative blood transfusion in MICS-BIMA group seemed less, though no significant difference was performed between the two groups[(0.5±1.2)U vs.(1.1±2.0)U, t=10.0, P=0.072]. There were no differences in MACCE, perioperative IABP use and wound infection between the two groups( P>0.05). In MICS-BIMA group, 40 IMA grafts from 20 patients were performed, with 8 in-situ grafts and 12 Y-graft. Except from LAD in all cases, 10 Diagonals, 4 RAMUSs, 4 OM/LCXs, 1 RCA and 1 PDA were bypassed. Operative angiography by CT scan showed a good patency of BIMA in MICS-BIMA group. Conclusion:Minimally invasive coronary surgery with bilateral internal mammary artery is a safe and effective approach for 2-vessel disease. Satisfied early clinical outcomes were shown in the study.

Objective To study the value of parameters of Transit-Time Flow Measurement(TTFM) as predictor and judge for one-year after coronary artery bypass transplantation in patency rate of vessel grafts .Methods Parameters of quantity of blood flow and the value of PI( pulsation index) were measured and recorded by intraoperative TTFM in CABG .One-year of follow-ing up, coronary artery CT or coronary angiography examination were accomplished.Results Our study include artery grafts 46(including LIMA grafts 44, RIMA graft 1 and radial artery graft 1) with the patency rate of grafts 91.3％ for one-year after CABG, and vein grafts 48 with the patency rate of grafts 66.7％ .There exists statistically significant difference between above two groups(P<0.001).High PI is independent risk factor(P =0.037) in patency rate of vessel grafts for one-year after CABG by binary logistics regression analysis but no other parameters have the statistical significance (P>0.05) .The results from logistic regression were summarized using the area under the ROC curve(AUC) .The pulsation index has been found hav-ing moderate judgment ability(P=0.016) for the dysfunction of grafts after CABG for one-year with optimal cut-off value of 2. 45.Conclusion Intraoperative high value of PI is the independent risk factor in patency rate of vessel grafts for one-year after CABG.