[Treatment of intrauterine adhesions in rats with hypoxia-cultured BMSC-derived exosomes].

Objective: To perform intrauterine adhesion modeling, and to investigate the repair effect of hypoxic treated bone marrow mesenchymal stem cells (BMSC) and their derived exosomes (BMSC-exo) on endometrial injury. Methods: BMSC and their exosomes BMSC-exo extracted from rats' femur were cultured under conventional oxygen condition (21%O2) or hypoxia condition (1%O2). Intrauterine adhesion modeling was performed on 40 healthy female SD rats by intrauterine injection of bacterial lipopolysaccharide after curettage. On the 28th day of modeling, 40 rat models were randomly divided into five groups, and interventions were performed: (1) NC group: 0.2 ml phosphate buffered solution was injected into each uterine cavity; (2) BMSC group: 0.2 ml BMSC (1×106/ml) with conventional oxygen culture was injected intrauterine; (3) L-BMSC group: 0.2 ml of hypoxic cultured BMSC (1×106/ml) was injected intrauterine; (4) BMSC-exo group: 0.2 ml of BMSC-exo cultured with conventional oxygen at a concentration of 500 µg/ml was injected into the uterine cavity; (5) L-BMSC-exo group: 0.2 ml hypoxic cultured BMSC-exo (500 µg/ml) was injected intrauterine. On the 14th and 28th day of treatment, four rats in each group were sacrificed by cervical dislocation after anesthesia, and endometrial tissues were collected. Then HE and Masson staining were used to observe and calculate the number of glands and fibrosis area in the endometrium. The expressions of angiogenesis related cytokines [vascular endothelial growth factor A (VEGFA) and CD31], and fibrosis-related proteins [collagen-Ⅰ, collagen-Ⅲ, smooth muscle actin α (α-SMA), and transforming growth factor ß1 (TGF-ß1)] in endometrial tissues were detected by western blot. Results: (1) HE and Masson staining showed that the number of endometrial glands in L-BMSC group, BMSC-exo group and L-BMSC-exo group increased and the fibrosis area decreased compared with NC group on the 14th and 28th day of treatment (all P<0.05). Noteworthily, the changes of L-BMSC-exo group were more significant than those of BMSC-exo group (all P<0.05), and the changes of BMSC-exo group were greater than those of BMSC group (all P<0.05). (2) Western blot analysis showed that, compared with NC group, the expressions of collagen-Ⅲ and TGF-ß1 in BMSC group, L-BMSC group, BMSC-exo group and L-BMSC-exo group decreased on the 14th and 28th day of treatment (all P<0.05). As the treatment time went on, the expressions of fibrosis-related proteins were different. Compared with BMSC group, the expressions of collagen-Ⅲ, α-SMA and TGF-ß1 in the BMSC-exo group and L-BMSC group decreased on the 28th day (all P<0.05). Moreover, the expressions of collagen-Ⅲ and TGF-ß1 in L-BMSC-exo group were lower than those in BMSC-exo group on the 28th day (all P<0.05). And the expressions of collagen-Ⅰ, α-SMA and TGF-ß1 in L-BMSC-exo group were lower than those in L-BMSC group on the 28th day (all P<0.05). (3) The results of western blot analysis of VEGFA and CD31 showed that, the expressions of VEGFA and CD31 in BMSC group, L-BMSC group, BMSC-exo group and L-BMSC-exo group increased on the 14th and 28th day of treatment compared with NC group (all P<0.05). Treatment for 28 days, the expressions of VEGFA and CD31 in BMSC-exo group and CD31 in L-BMSC group were higher than those in BMSC group (all P<0.05). Moreover, the expressions of VEGFA and CD31 in L-BMSC-exo group were higher than those in BMSC-exo group and L-BMSC group on the 28th day (all P<0.05). Conclusions: Treatment of BMSC and their exosomes BMSC-exo with hypoxia could promote endometrial gland hyperplasia, inhibit tissue fibrosis, and further repair the damaged endometrium in rats with intrauterine adhesion. Importantly, hypoxic treatment of BMSC-exo is the most effective in intrauterine adhesion rats.

[Multivariate analysis of the effect of two liver resection methods on the survival outcome of patients with intrahepatic cholangiocarcinoma].

Objective: To investigate the effect of anatomical hepatectomy and non-anatomic hepatectomy in the treatment of elderly patients with intrahepatic cholangiocarcinoma (IHCC) and their impact on survival outcomes. Methods: In this study, a retrospective method was used to select elderly patients with IHCC who were surgically treated in Shangqiu First People's Hospital from April 2014 to April 2018, and were divided into anatomic resection group and non-anatomical resection group according to the surgical methods they received.The factors affecting the survival outcome of IHCC in the two liver resection methods were analyzed and compared, as well as the effects of liver cirrhosis rate, TNM stage, ascites rate, lymph node metastasis rate, and vascular invasion rate on survival. Results: A total of 181 cases were included in this study, including 87 cases in the anatomical resection group, with 54 males and 33 females, aged (71.4±5.2) years old;There were 94 cases in the non-anatomical resection group, including 49 males and 45 females, aged (70.8±4.8) years.The 3-year survival rate of the anatomical resection group was 41.4% (36/87), which was higher than that of the non-anatomical resection group (25.5% (24/94), the difference was statistically significant (P<0.05);The median survival time of the anatomic resection group was longer than that of the non-anatomical resection group, and the difference was statistically significant P<0.05;The patient's TNM stage was stage III [OR (95%CI): 2.168 (1.245-3.776)], lymph node metastasis [1.664 (1.087-2.545)], and vascular invasion [1.883 (1.167-3.038)] was an independent risk factor for death 3 years after surgery (P<0.05), The patient's anatomical liver resection was a protective factor for the 3-year follow-up survival (P<0.05). Conclusion: The postoperative survival of elderly patients with IHCC is affected by many factors, but anatomic liver resection is beneficial to prolong the survival time of patients.

[Study on the differentiation of rat bone marrow mesenchymal stem cells into chondrocytes induced by TGF-beta 3].

Objective: To explore the methods for the differentiation of rat bone marrow mesenchymal stem cells (BMMSCs) into chondrocytes induced by transforming growth factor (TGF)-beta 3 in vitro, and search for a reliable seed cell for the regeneration and repair of articular cartilage in osteoarthritis. Methods: SD rat femoral and tibial BMMSCs were cultured by the whole bone marrow adherent method, and then the purity was identified by flow cytometry. P3 generation cells were induced and differentiated into chondrocytes by the induction of differentiation medium containing TGF-beta 3, and cell chondrogenic differentiation ability at different induction time points was detected. Results: The primary and passage cultured BMMSCs were spindle-shaped, and partly triangular. After passage, the proliferation rate of P3 generation cells was fast, like the growth of fish shoal or eddy. After chondrogenic induction, the cells were polygonal and triangular in the form of cluster growth, which was similar to chondrocyte morphology, and the cell proliferation was decreased. Immunofluorescence staining and Western blotting method showed that the cells had a large number of col Ⅱ fluorescent expression, and cells and extracellular matrix was stained blue by Alcian blue staining, with no significant difference at day 7 and day 14. After using Wnt signal blocker, col Ⅱ protein expression was significantly reduced, with statistically significant difference (P<0.05). Conclusion: TGF-beta 3 can rapidly induce the differentiation of BMMSCs into cartilage cells, which provides a good carrier for BMMSCs transplantation and the repair of articular cartilage, and thus to treat osteoarthritis.

[Regulatory role of galanin on prolactin and beta-endorphin release from anterior pituitary lobe of rat].

The role of brain-gut peptide galanin in the regulation of prolactin (PRL) and beta-endorphin (beta-EP) release from anterior pituitary lobe was studied in vivo in conscious male rats and in vitro with cultured anterior pituitary cells of the rat. Galanin (1 microgram or 3 micrograms/rat) injected into the third cerebral ventricle of rats produced highly significant, dose-related increases of PRL resting secretion, but did not alter resting secretion of beta-EP and restraint stress-induced release of PRL and beta-EP. However, galanin (0.05, 0.5 and 1.0 micrograms/5 x 10(5) cells.ml-1) induced highly significant, dose-related increase of beta-EP secretion from dispersed anterior pituitary cells, but failed to alter significantly PRL secretion from the cultured cells. These results indicate that central galanin has a stimulatory role in pituitary PRL resting secretion via the hypothalamus, whereas peripheral galanin stimulates beta-EP secretion only via direct action of this peptide in anterior pituitary cells.

[Pancreatic polypeptide in the control of beta-endorphin and prolactin release from rat anterior pituitary in vivo and in vitro].

This study was designed to examine the possible effects of pancreatic polypeptide (PP) on beta-endorphin (beta-EP) and prolactin (PRL) release from rat anterior pituitary in vivo and in vitro. Injection of 0.5 microgram or 2.0 micrograms PP into the third ventricle of the brain (3rd v.i.) produced a significant decrease of the beta-EP and PRL resting secretion. 0.5 microgram PP (3rd v.i.) did not affect restraint stress-induced release of beta-EP, but partially lowered stress induced release of PRL. 2.0 micrograms PP (3rd v.i.) partially reduced restraint stress-induced release of beta-EP and completely suppressed stress-induced release of PRL. In order to investigate a possible direct action of PP on beta-EP and PRL secretion from the anterior pituitary gland, we incubated dispersed anterior pituitary cells with synthetic PP (0.05, 0.625 and 1.00 micrograms) for 1 n, the secretion of beta-EP was not affected at any dosage tested, but 0.625 and 1.00 micrograms PP significantly decreased the PRL secretion. These data indicate that PP may have an inhibitory role in the control of beta-EP secretion at the level of the hypothalamus, and an inhibitory role in the control of PRL secretion at the level of either hypothalamus or anterior pituitary.