Objective To investigate the effect of dynamic compression and rotation motion on chondrogenesis of the 3rd passage cell-loaded three-dimensional scaffold in a joint-specific bioreactor in vitro so as to provide theoretical basis of the autologous chondrocyte transplantation in clinical practice. Methods Primary chondrocytes were isolated and cultured from the knee cartilage of 3-4 months old calves. The 3rd passage cells were seeded onto fibrin-polyurethane scaffolds (8 mm × 4 mm). Experiment included 5 groups: unloaded culture for 2 weeks (group A), direct load for 2 weeks (group B), unloaded culture for 4 weeks (group C), direct load for 4 weeks (group D), and unload for 2 weeks followed by load for 2 weeks (group E). The cell-scaffold was incubated in incubator (unload) or in a joint-specific bioreactor (load culture). At different time points, the samples were collected for DNA and glycosaminoglycan (GAG) quantification detect; mRNA expressions of chondrogenic marker genes such as collagen type I, collagen type II, Aggrecan, cartilage oligomeric matrix protein (COMP), and superficial zone protein (SZP) were detected by real-time quantitative PCR; and histology observations were done by toluidine blue staining and immunohistochemistry staining. Results No significant difference was found in DNA content, GAG content, and the ratio of GAG to DNA among 5 groups (P gt; 0.05). After load, there was a large number of GAG in the medium, and the GAG significantly increased with time (P lt; 0.05). The mRNA expression of collagen type I showed no significant difference among 5 groups (P gt; 0.05). The mRNA expression of collagen type II in group B was significantly increased when compared with group A (P lt; 0.01), and groups D and E were significantly higher than group C (P lt; 0.01); the mRNA expression of Aggrecan in groups D and E were significantly increased when compared with group C (P lt; 0.01), and group E was significantly higher than group D (P lt; 0.01); the mRNA expression of COMP in group B was significantly increased when compared with group A (P lt; 0.01), and group E was significantly higher than group C (P lt; 0.01); and the mRNA expression of SZP in group E was significantly increased when compared with groups C and D (P lt; 0.05). The toluidine blue staining and immunohistochemistry staining displayed that synthesis and secretion of GAG could be enhanced after load; no intensity changes of collagen type I and collagen type II were observed, but intensity enhancement of Agrrecan was seen in groups D and E. Conclusion Different dynamic loads can promote chondrogenesis of the 3rd passage chondrocytes. Culture by load after unload may be the best culture for chondrogenesis, while the 3rd passage chondrocytes induced by mechanical load hold less capacity of chondrogenesis.
Objective To explore a better method in obtaining iris pigment epithelium(IPE) specimen for autologous transplantation in rabbits． Methods IPE was obtained from 20 black rabbits with method A，i.e.surgical peripheral iridectomy at 12:00 position obtaining a triangle iris tissue with the hemline of 4-5 mm in left eyes，and method B,i.e.surgical peripheral iridectomy at 11:00 and 1:00 positions obtaining two triangle iris tissues with the hemlines of 2-2.5 mm in right eyes . The IP E cells were isolated precisely with enzyme microdissection-enzyme isolation method, cultured in vitro, observed with light and electronic microscope, and ident ified with immunocytochemical staining.ResultsThe success ra te of cells culture were 65% for method A and 95% for method B. After 3-4 generations of culturing，the amount of IPE cells was enough for transplantation, and most of the functions of primary clutured IPE cells were kept still. Viability of IPE cells was 85%-93%. Conclusion The success rate of cells culture for method B is higher than that for method A. The third generation of cultured cells is available for autologous transplantation.(Chin J Ocul Fundus Dis，2003，19：201-268)
Objective To analyze the learning curve of unilateral biportal endoscopic lumbar interbody fusion (UBE-LIF). Methods Fifty-five patients with single-segment lumbar degenerative disease treated with UBE-LIF between December 2020 and February 2022 were selected as the research subjects. The patients were grouped according to the operation sequence, the first 27 cases were in the early group, and the last 28 cases were in the late group. There was no significant difference between the two groups in age, gender, disease type, and surgical segment distribution (P>0.05). The operation time, the amount of hemoglobin loss (the difference between 1 day before operation and 3 days after operation), the hospital stay after operation, and the incidence of perioperative complications were recorded; the learning curve of UBE-LIF was analyzed by log-curve regression analysis. Results All the operations were successfully completed without changing to other operations. The operation time, the amount of hemoglobin loss, and hospital stay in the early group were significantly more than those in the late group (P<0.05). Complications occurred in 2 cases (7.4%) in the early group, including 1 case of dural tear during operation and 1 case of epidural hematoma after operation, and 1 case (3.6%) with transient radiculitis in the late group. There was no significant difference in the incidence of complications between the two groups (P=0.518) . The log-curve regression analysis showed that the operation time decreased significantly with the increase of the number of patients (P<0.05). The operation time tended to be stable after the surgeon completed 17 cases. ConclusionFor single-level lumbar degenerative disease, the operation time of UBE-LIF can decrease gradually with the increase of the number of patients, and tend to be stable after 17 cases.
Objective To investigate preoperative blood neutrophil-to-lymphocyte ratio （NLR） in patients with gastrointestinal stromal tumor （GIST） and analyze the relationship of the NLR to prognosis. Methods The data of NLR of peripheral blood samples on 3d before surgery and the results of immunohistochemistry of 42 patients with GIST were analyzed respectively，the relation between the NLR and the prognosis of patients with GIST was understood by the survival analysis. Results The patients with high NLR （NLR≥2.5） was found in 22 cases， low NLR （NLR＜2.5） in 20 cases.The NLR was related to mitotic figures （χ2=9.45，P=0.002） and tumor size （P=0.041）. The 3-year survival rate of the patients with high NLR was shorter than that of the low NLR （χ2=5.44，P=0.022）. The 3-year survival rate was associated with NLR，mitotic figures，and tumor size （P＜0.05） in univariate analysis. The NLR and mitotic figures were independent prognostic indicators of 3-year survival （P=0.018，P=0.000） in Cox multivariate analysis. Conclusion Blood NLR and mitotic figures have some predictive value for the prognosis of patients with GIST.