To investigate the inhibition of HBV expression by multi-target ribozyme-expressing adenovirus vector in vivo, the ribozyme and muta-ribozyme were cloned into adenovirus vector pAd-CMV. By using the adenovirus expression system, the two plasmids were co-transfected into 293 cell line with adenovirus gene recombinant plasmids pJM17. The recombinant adenovirus were detected by plaque assay. Then the recombinant adenovirus infected the 2.2.15 cells. Using ELISA, dot-bolt hybridization and image analysis system, the inhibition of HBV expression was detected. The ribozyme and muta-ribozyme were cloned into the adenovirus vector pAd-CMV. The recombinant adenovirus could infect 2.2.15 cells and express the ribozyme. The ribozyme could decrease the expression of HBeAg by 87.1%. All these results strongly indicate that ribozyme can inhibit HBV expression in the cell, which represents a potential gene therapy for HBV infection.

Objective:To explore the mediating role of mindfulness in part-time nursing faculty between intrinsic motivation and work engagement.Methods:A total of 376 part-time nursing faculty were investigated with Work Preference Inventory, Five Facet Mindfulness Questionnaire-Short Form and Utrecht Work Engagement Scale. SPSS 25.0 statistical software was used to perform t test on the data. Results:The intrinsic motivation of part-time nursing faculty positively predicted work engagement ( β=0.74, P<0.001). The positive predictive effect of intrinsic motivation on job engagement was weaker than before ( β=0.55, P<0.001). Mindfulness as an intermediary variable reduced the positive predictive effect of intrinsic motivation on work engagement from 0.979 to 0.504. The indirect effect of intrinsic motivation on work engagement through mindfulness was significant (path coefficient=0.50, P<0.001), and the intermediary effect accounted for 48.51% of the total effect. Conclusion:Mindfulness plays an intermediary role between intrinsic motivation and work engagement of part-time nursing faculty. Paying attention to the mindfulness level of part-time nursing faculty plays a positive role in improving their work engagement level.

BACKGROUNDTo investigate the biological effects of anti VEGF₁₆₅ ribozyme on human lung adenocarcinoma cell.

METHODSHammerhead ribozyme (VRz) against VEGF₁₆₅ gene transcripts (site 212) and its paired mutant ribozyme (mVRz) were designed and synthesized, and the cleavage activity of the ribozymes on target RNA in a cell-free system was observed. The replication-incompetent adenovirus-mediated eukaryotic expression vectors (rpAdVRz) containing VRz and mVRz gene were constructed and identified. Then the human lung adenocarcinoma cells (A549) were infected with recombinant adenovirus. The biological characteristics of A549 cell before and after infection in vitro were inspected by Northern blot, laser confocal imaging system analysis, flow cytometry and transmission electron microscopy.

RESULTSVRz specifically and efficiently cleaved the VEGF₁₆₅ mRNA. The rpAdVRz was successfully constructed and infected A549 cell. The level of VEGF₁₆₅ expression decreased 87% in rpAdVRz infected cells compared with the other groups, but their biological characteristics were not influenced by the expression of the exogenous gene.

CONCLUSIONSThe adenovirus mediated hammerhead ribozyme against VEGF₁₆₅ can significantly decrease the expression of VEGF₁₆₅. This provides an experimental basis for human lung cancer gene therapy with antiangiogenesis method.

Objective:To introduce a novel anatomical supra-pectineal quadrilateral surface buttress plate, and explore its advantages, disadvantages and clinical efficacy in the treatment of complex acetabular fractures.Methods:Data of eighteen cases of acetabular fractures treated in our department from March 2017 to March 2019were retrospectively analyzed. There were 15 males and 3 females with an average age of 49.6 years (range, 27-62 years). According to the Letournel-Judet’s classification, there were 3 cases of anterior column fractures (involving quadrilateral surface), 2 cases of "T" shaped, 8 of cases anterior column and posterior hemi-transverse (ACPHT) and 5 cases of double columns fractures. The time from injury to operation was 5-14 days (mean, 7 days). Before operation, all patients were taken pelvic anterior-posterior X-ray and CT scan. All patients were adopted supra-ilioinguinal approach and fixed by supra-pectineal quadrilateral surface buttress plate. Surgical time, intraoperative blood loss and postoperative complications were collected. After operations all patients were taken pelvic X-ray in anterior-posterior and Judet position, as well as CT scan. The radiographic quality of reduction was evaluated by Matta criteria and hip functions were evaluated by Matta Modified Merle d’Aubigné-Postel scoring system.Results:The average surgical time was 130 min (range, 100-200 min). The average blood loss was 560 ml (range, 400-900 ml). An additional shaping was unnecessary in 18 patients who were implanted with the anatomical supra-pectineal quadrilateral surface buttress plate and all patients were operated successfully. All patients were followed up for an average time of 9 months (range, 6-15 months). The quality of fracture reduction was evaluated according to Matta standard, of which there were 11 cases excellent, 4 cases good, and 3 cases poor, with an excellent rate of 83.3% (15/18). At the latest follow-up, the function of the hip joint was evaluated according to the Matta Modified Merle d’Aubigné-Postel scoring system for a mean of 15.8 scores (range, 12-18 scores), of which there were 10 cases excellent, 6 cases good, 2 cases fair, with an excellent rate of 88.9% (16/18). All patients recovered well, and there were 2 cases of lateral femoral cutaneous nerve injury postoperative who had recovered one month later. There was no other postoperative complications.Conclusion:The anterior and posterior column and quadrilateral surface could be stabilized simultaneously by the anatomical supra-pectineal quadrilateral surface buttress plate without intraoperative shaping. Moreover, it is more likely to obtain a satisfactory clinical outcome by using this special plate with lower complications and rigid fixation. Therefore, it is a kind of internal fixation method worthy of promotion in the treatment of complex acetabular fractures.

Objective:To investigate the clinical features and therapeutic efficacy of patients with hereditary leiomyomatosis and renal cell carcinoma(RCC) syndrome-associated RCC (HLRCC-RCC) in China.Methods:The clinical data of 119 HLRCC-RCC patients with fumarate hydratase (FH) germline mutation confirmed by genetic diagnosis from 15 medical centers nationwide from January 2008 to December 2021 were retrospectively analyzed. Among them, 73 were male and 46 were female. The median age was 38(13, 74) years. The median tumor diameter was 6.5 (1.0, 20.5) cm. There were 38 cases (31.9%) in stage Ⅰ-Ⅱand 81 cases (68.1%) in stage Ⅲ-Ⅳ. In this group, only 11 of 119 HLRCC-RCC patients presented with skin smooth muscle tumors, and 44 of 46 female HLRCC-RCC patients had a history of uterine fibroids. The pathological characteristics, treatment methods, prognosis and survival of the patients were summarized.Results:A total of 86 patients underwent surgical treatment, including 70 cases of radical nephrectomy, 5 cases of partial nephrectomy, and 11 cases of reductive nephrectomy. The other 33 patients with newly diagnosed metastasis underwent renal puncture biopsy. The results of genetic testing showed that 94 patients had FH gene point mutation, 18 had FH gene insertion/deletion mutation, 4 had FH gene splicing mutation, 2 had FH gene large fragment deletion and 1 had FH gene copy number mutation. Immunohistochemical staining showed strong 2-succinocysteine (2-SC) positive and FH negative in 113 patients. A total of 102 patients received systematic treatment, including 44 newly diagnosed patients with metastasis and 58 patients with postoperative metastasis. Among them, 33 patients were treated with tyrosine kinase inhibitor (TKI) combined with immune checkpoint inhibitor (ICI), 8 patients were treated with bevacizumab combined with erlotinib, and 61 patients were treated with TKI monotherapy. Survival analysis showed that the median progression-free survival (PFS) of TKI combined with ICI was 18 (5, 38) months, and the median overall survival (OS) was not reached. The median PFS and OS were 12 (5, 14) months and 30 (10, 32) months in the bevacizumab combined with erlotinib treatment group, respectively. The median PFS and OS were 10 (3, 64) months and 44 (10, 74) months in the TKI monotherapy group, respectively. PFS ( P=0.009) and OS ( P=0.006) in TKI combined with ICI group were better than those in bevacizumab combined with erlotinib group. The median PFS ( P=0.003) and median OS ( P=0.028) in TKI combined with ICI group were better than those in TKI monotherapy group. Conclusions:HLRCC-RCC is rare but has a high degree of malignancy, poor prognosis and familial genetic characteristics. Immunohistochemical staining with strong positive 2-SC and negative FH can provide an important basis for clinical diagnosis. Genetic detection of FH gene germ line mutation can confirm the diagnosis. The preliminary study results confirmed that TKI combined with ICI had a good clinical effect, but it needs to be confirmed by the results of a large sample multi-center randomized controlled clinical study.