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In this study, high-throughput sequencing technology and statistical analysis were employed to explore the effects of adding green tea at proportions of 10%, 20%, and 30% on physicochemical characteristics, bacterial community diversity and succession of Daqu during fermentation compared with pure wheat Daqu (control). The results showed that the addition of green tea significantly increased the bacterial community diversity of Daqu and improved its saccharification capacity, moisture content and acidity. However, the increase in tea addition did not exhibit a proportional relationship with the community diversity. The unique evolutionary structure of the bacterial communities in different types of Daqu was found to be the major factor affecting the physicochemical characteristics of Daqu. During the fermentation process, all four Daqu exhibited differentiation in bacterial community succession starting from the thermophilic stage, thereby leading to the formation of their unique community structure at the end of the fermentation process. The phyla Proteobacteria, Firmicutes, Bacteroidota, and Actinobacteria showed varying influences on the community succession of the four Daqu samples, thus making each of them have different physicochemical characteristics. Specifically, Pseudomonas was an important bacterial genus contributing to the physicochemical characteristics of the control and 10% tea Daqu, while Weissella and Pediococcus, were important contributors to the physicochemical characteristics of the 20% tea Daqu. Bacillus was found to be a significant bacterial genus contributing to the physicochemical characteristics of the 30% tea Daqu. This study provides a theoretical basis for process optimization in the production of tea-added Daqu and for the development of multi-raw material fermentation starters. It also has implications for improving the preparation process of Daqu and the application of tea in Daqu making.

Synthetic Biology, as an emerging discipline, has gained widespread attention and is developing rapidly, profoundly impacting the fields of life sciences and biotechnology. Concurrently, as emerging engineering education programs take shape, accelerated cultivation of multifaceted innovative talents represents a new mission and imperative for higher education in China. In the context of the flourishing development of Synthetic Biology, East China University of Science and Technology has established a curriculum cluster in Synthetic Biology, focusing on microbiological drug discovery and biomanufacturing. The teaching team initially reviewed the curriculum system related to Synthetic Biology and its upstream and downstream courses. Subsequently, they expanded the core courses in Synthetic Biology, creating a curriculum cluster that encompasses not only the theoretical foundations and cutting-edge technologies but also integrates with related disciplines. Moreover, the curriculum cluster leverages lectures from renowned domestic and international professors in the State Key Laboratory of Bioreactor Engineering, and harnesses the rich resources of the Program of Introducing Talents of Discipline to Universities (the "111 plan"), aiming to enhance students' innovation capabilities. With the support of this curriculum cluster and teaching team, undergraduate students actively participate in international Synthetic Biology competitions like international genetic engineering machine competition (iGEM), consistently achieving gold awards. Furthermore, many students have applied for patents and made contributions to research paper publications. This work stands as a valuable exemplar for cultivating multifaceted talents with exceptional innovative capabilities.

Objective: To construct a eukaryotic expression vector of bromodomain-containing protein 7 (BRD7) with deletion of bromodomain (BRD7△brd) using the homologous recombination and reverse PCR amplification techniques., Methods: The linear DNA fragments of bromodomain-deleted mutation of BRD7 (pIRES2-EGFP- 3Flag/BRD7△brd) were amplified by one pair of reverse PCR primers using high-fidelity enzyme, and then these fragments were transformed into E.coli to obtain the eukaryotic expression vector expressing BRD7△brd protein based on homologous recombination and plasmid cyclization., Results: Bromodomain-deleted clones were identified by digestion with restrictive enzymes, and then the sequence and protein expression were further confirmed by sequencing and Western blot assays. The results suggest that pIRES2-EGFP-3Flag/BRD7△brd was successfully constructed., Conclusion: We establish a simple and quick method to construct plasmids with pIRES2-EGFP- 3Flag/BRD7△brd using reverse PCR amplification and homologous recombination techniques. We also found that the concentration of template in PCR reaction system is one of the critical factors that affect the rate of homologous recombination. Of all, this improved technique could be widely used in the construction of gene mutations.

Objective: To evaluate the feasibility, safety and short-term clinical outcomes of robot-assisted minimally invasive esophagectomy (RAMIE)., Methods: Clinical data of 17 patients with esophageal cancer who received RAMIE between April 2016 and July 2016 were analyzed retrospectively., Results: The age of the patients ranged from 44 to 83. Six patients received neoadjuvant radiochemotherapy while 11 patients underwent surgery alone. All patients were performed by the robot-assisted thoraco-laparoscopic minimally invasive esophagectomy. In-hospital mortality was 0%. None was converted to open transthoracic or laparotomy approach. In the neoadjuvant radiochemotherapy group, 3 patients received pathological complete response while 2 patients were stage II(A and 1 patient was stage II(B. In the surgery alone group, 1 patient was stage I(A, 3 patients were stage II(A, 5 patients were stage II(B, 1 patient was stage III(A and 1 patient was stage III(B. The mean operation time was 195 minutes (range 145 to 305 minutes). The mean blood loss was 60 ml (range 30 to 200 ml). Mean lymph node harvest was 28 nodes. The rate of radical resection was 100%. Median ICU stay was 4.5 days (range 1 to 36 days), and median overall postoperative hospital stay was 15.2 days(range 9 to 45 days). Postoperative complication occurred in 4 (23.5%) patients, including 3 (17.6%) of lung lesion, 2 (11.8%) of hoarseness, 1 (5.9%) of chylothorax, while no anastomotic leakage and arrhythmia was observed., Conclusion: RAMIE for esophageal cancer is feasible and safe with favorable early outcomes.