OBJECTIVE: To explore the degree, mechanism and clinical significance of three-dimensional tumor microvascular architecture phenotype heterogeneity (3D-TMAPH) in non-small cell carcinoma (NSCLC). METHODS: Twenty-one samples of solitary pulmonary nodules were collected integrally. To establish two-dimensional tumor microvascular architecture phenotype (2D-TMAP) and three-dimensional tumor microvascular architecture phenotype (3D-TMAP), five layers of each nodule were selected and embedded in paraffin. Test indices included the expressions of vascular endothelial growth factor (VEGF), proliferating cell nuclear antigen (PCNA), EphB4, ephfinB2 and microvascular density marked by anti-CD34 (CD34-MVD). The degrees of 3D-TMAPH were evaluated by the coefficient of variation and extend of heterogeneity. Spearman rank correlation analysis was used to investigate the relationships between 2D-TMAP, 3D-TMAP and clinicopathological features. RESULTS: 3D-TMAPH showed that 2D-TMAP heterogeneity was expressed in the tissues of NSCLC. The heterogeneities in the malignant nodules were significantly higher than those in the active inflammatory nodules and tubercular nodules. In addition, different degrees of heterogeneity of CD34-MVD and PCNA were found in NSCLC tissues. The coefficients of variation of CD34- MVD and PCNA were positively related to the degree of differentiation (all P<0.05), but not related to the P-TNM stages, histological type or lymphatic metastasis (all P>0.05). The level of heterogeneity of various expression indexes (ephrinB2, EphB4, VEGF) in NSCLC tissues were inconsistent, but there were no significant differences in heterogeneity in NSCLC tissues with different histological types (P>0.05). CONCLUSION 3D-TMAPH exists widely in the microenvironment during the genesis and development of NSCLC and has a significant impact on its biological complexity.
Adult ; Aged ; Capillaries ; ultrastructure ; Carcinoma, Non-Small-Cell Lung ; blood supply ; Ephrin-B2 ; metabolism ; Female ; Humans ; Lung Neoplasms ; blood supply ; Male ; Middle Aged ; Neovascularization, Pathologic ; pathology ; Phenotype ; Proliferating Cell Nuclear Antigen ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

Objective:To investigate the correlation between time within the glucose target range(TIR) and hyperuricemia(HUA) in patients with type 2 diabetes mellitus(T2DM).Methods:A total of 215 patients with T2DM in the First Affiliated Hospital of Bengbu Medical College from June 2021 to May 2022 were selected and divided into HUA group and non-HUA group according to serum uric acid level. The clinical characteristics and biochemical indicators of the patients were collected. The association of 72 h glucose monitoring system(FGMS) related indicators TIR, mean blood glucose fluctuation range(MAGE), blood glucose variability(CV), blood glucose standard deviation(SDBG), and mean blood glucose(MBG) with serum uric acid level was analyzed. The influencing factors of T2DM combined with HUA were analyzed with binary logistic regression, and receiver operating characteristic(ROC) curve was drawn to evaluate their predictive values.Results:TIR of HUA group was significantly decreased compared with non-HUA group, while HbA 1C, MAGE, CV, SDBG, and MBG were increased( P<0.001). Spearman correlation analysis showed that serum uric acid levels were negatively correlated with TIR, but positively correlated with MAGE, CV, SDBG, and MBG( P<0.001). After dichotomous logistic regression analysis, TIR was found to be an independent protective factor for T2DM with HUA. The ROC curve results showed that the area under the curve(AUC) of TIR for predicting HUA in T2DM was 0.856(95% CI 0.803-0.909, P<0.001), with the best cut-off value being 64.5%, the sensitivity being 76.8%, and the specificity being 90.3%. Conclusion:TIR in patients with T2DM combined with HUA was significantly decreased. TIR is an independent protective factor for T2DM combined with HUA, and TIR shows a certain predictive value for T2DM combined with HUA.

Objective : To study the effect of leucine rich repeat kinase 2 ( LRRK2) on pain sensitivity in neuro- pathic pain (NP) rats and explore its possible mechanism． Methods : 48 SD rats were randomly divided into four groups : sham surgery (Sham) ，model，LRRK2 inhibitor(MLi-2) ，and LRRK2 inhibitor + p38 mitogen activated pro- tein kinase (MAPK) agonist (MLi-2 + Anisomycin) ，with 12 rats in each group．The NP rat model was induced by chronic constriction injury ( CCI) of the sciatic nerve．Intrathecal injection of MLi-2 ( 1 mg / kg，10 μl) or Anisomy- cin (20 μmol / L，10 μl) was started from the 8 th day after surgery，once a day for 7 consecutive days．Pain sensi- tivity tests were conducted before surgery (day 0) and on postoperative days 7 and 14，respectively．The changes in mechanical withdrawal threshold (MWT) and paw withdraw thermal latency (PWTL) were analyzed in each group of rats．ELISA was used to detect the levels of interleukin-1 β (IL-1 β) ，IL-6 and tumor necrosis factor-α (TNF-α) in the dorsal horn of the rat spinal cord．Nissl staining was used to observe the pathological changes of neurons in rat spinal cord tissue．Immunofluorescence staining was used to observe the expression levels of ionized calcium-binding adapter molecule-1 (Iba-1) ，a marker of microglia in the spinal cord of rats ．Western blot was used to detect theprotein expression levels of LRRK2，p-p38 mitogen activated protein kinase (MAPK) ，p38 MAPK，and Iba-1 in the dorsal horn of the rat spinal cord. Results : Compared with the sham group，the model group showed a significant decrease in MWT and PWTL in the right hind limb of rats (P＜0. 01) ．The levels of IL-1，IL-6，and TNF in the spi- nal dorsal horn tissue，as well as the expression levels of LRRK2，Iba-1 proteins and p-p38 MAPK / p38 MAPK pro- tein ratio significantly increased (P＜0. 01) ．The proportion of Iba-1 positive cells in the spinal cord tissue signifi- cantly increased (P＜0. 01) ，while Nissl bodies were significantly reduced (P＜0. 01) ．Compared with the model group，the MLi-2 group showed a significant increase in MWT and PWTL in the right hind limb of rats (P＜0. 01) ， a significant increase in Nissl bodies (P＜0. 01) ，a significant decrease in the proportion of Iba-1 positive cells in the spinal cord tissue (P＜0. 01) ，and a significant decrease in the levels of IL-1，IL-6，and TNF and the expression levels of LRRK2，Iba-1 proteins and p-p38 MAPK / p38 MAPK protein ratio (P＜0. 01) ．However，Anisomychin in- tervention could activate the p38 MAPK signaling pathway and partially reverse the beneficial effects of MLi-2 on pain sensitivity and neuroinflammation in rats with neuropathic pain． Conclusion Inhibiting the expression of LRRK2 can alleviate pain sensitivity in NP rats induced by microglia activation mediated neuroinflammation，and its mechanism of action may be related to regulating the p38 MAPK signaling pathway.