Clinical application of modified Crain classification in the Design of Anterior Cruciate Ligament Reconstruction with remnant preservation.

BACKGROUND: To investigate the clinical application of modified Crain classification in anterior cruciate ligament (ACL) reconstruction (ACLR) with remnant preservation. METHODS: The subjects were 70 patients with ACL injury who underwent ACLR from May 2016 to June 2018, and their general data were recorded. They were randomly divided into modified remnant-preserved ACLR group (group M, n = 35) and non remnant-preserved ACLR group (group N, n = 35). ACLR program with remnant preservation was designed based on modified Crain classification in group M, while ACL remnants were completely cleaned during ACLR in group N. Subsequently, the two groups were compared in terms of operation time, complications, as well as Lysholm score, international knee documentation committee (IKDC) score and positive rate of Lachman test of knee joint before operation and at 3, 6 and 12 months after operation. RESULTS: Both the groups showed good postoperative efficacy, and none had complications like limited knee extension or cyclops lesion. The comparison results found that group M (72.49 ± 7.64 min) required longer operation time than group N (66.06 ± 6.37 min) (P < 0.05). Lysholm score and IKDC score at 3, 6 and 12 months after operation in the two groups were significantly higher than those before operation (P < 0.05); group M had higher Lysholm score and IKDC score at 3 months and 6 months after operation compared with group N (P < 0.05). Additionally, the positive rate of Lachman test at 3, 6 and 12 months after operation in both groups was significantly lower than that before operation (P < 0.05), but there was no significant difference between group M and group N. CONCLUSION: With the modified Crain classification, many remnant-preserved reconstruction techniques can be rationally used to completely preserve the remnant ligament tissue during operation and improve knee joint function and joint stability with few complications.

Microbial Cell Factory of Baccatin III Preparation in Escherichia coli by Increasing DBAT Thermostability and in vivo Acetyl-CoA Supply.

Given the rapid development of genome mining in this decade, the substrate channel of paclitaxel might be identified in the near future. A robust microbial cell factory with gene dbat, encoding a key rate-limiting enzyme 10-deacetylbaccatin III-10-O-transferase (DBAT) in paclitaxel biosynthesis to synthesize the precursor baccatin III, will lay out a promising foundation for paclitaxel de novo synthesis. Here, we integrated gene dbat into the wild-type Escherichia coli BW25113 to construct strain BWD01. Yet, it was relatively unstable in baccatin III synthesis. Mutant gene dbat S189V with improved thermostability was screened out from a semi-rational mutation library of DBAT. When it was over-expressed in an engineered strain N05 with improved acetyl-CoA generation, combined with carbon source optimization of fermentation engineering, the production level of baccatin III was significantly increased. Using this combination, integrated strain N05S01 with mutant dbat S189V achieved a 10.50-fold increase in baccatin III production compared with original strain BWD01. Our findings suggest that the combination of protein engineering and metabolic engineering will become a promising strategy for paclitaxel production.