Objective To investigate the developmental feasibility of early human fetal testes (<3 months) using xenografting technique and to acquire an accessible donor derivation that is essential for studying human germ cell development. Methods Nine testes from 10-13 weeks aborted fetus were grafted under the back skin of 6 castrated nude mice. Grafts were collected at different time point according to the growth of the donor tissues and the health condition of the recipients. Morphological and histological analyses were performed for the observation of the development of grafted immature testicular tissues. Results The mass of grafts was increased from about 5-7mg to 84.1mg (the biggest). Six of 9 testes were to be in developing. Histological observations showed a significant expansion of seminiferous tubules from (44.26±3.14)μm to (77.69±7.47)μm. Cells dispersedly distributed in seminiferous cords at the time of grafting migrated towards the basal part of seminiferous epithelium. Some germ cells with spermatogonium-like characteristics located on the basement membrane. Sertoli cells were in stages from immature into matured with abundant cytoplasm which were orderly arranged around spermatogonia forming a niche-like structure. Conclusion Testes from early aborted human fetus grafted under the back skin of castrated nude mice showed further development and therefore could be used as an easier accessible donor tissues for the investigation of human spermatogenetic mechanism.

Objective:Under the new development of breast cancer diagnosis and treatment technology, the relationship between nuclear morphology and different clinicopathological characteristics of breast cancer was analyzed, and its clinical significance was evaluated.Methods:In TCGA (the Cancer Genome Atlas) BRCA (fast invasive carcinoma) data, 443 patients were divided into three groups according to the nuclear morphology: ⑴ the nuclear size and morphology were basically normal; ⑵ the nuclear size slightly increased; ⑶ the nuclear size slightly increased or the difference was obvious. Practical clinicopathological features including American Joint Committee on Cancer (AJCC) tumor stage, AJCC tumor score, AJCC lymph node score, estrogen receptor (ER) status by immunohistochemistry (IHC), progesterone receptor (PR) status by IHC, human epidermal growth factor receptor 2 (HER2) status by IHC and PAM50 subtypes are compared among groups defined by nuclear size. And nuclear size related morphometric parameters, including nuclear area, perimeter, roundness, width and height, are subsequently retrieved in 132 samples and analyzed for their relationship with clinicopathological features.Results:The nuclear size in breast cancer tissues was significantly correlated with survival, especially disease-free survival ( P=0.039). Other clinicopathological parameters, except AJCC T stage ( P=0.006), the immunohistochemical results of molecular markers ER ( P=0.002), PR ( P=0.047) and molecular typing of PAM50 ( P<0.001) were not significantly associated with the nuclear size of cancer tissues. The area, perimeter, roundness, length and width were correlated with each other; the roundness was the most stable parameter and negatively correlated with other parameters; the perimeter was the most sensitive index for identification, while roundness was not. Conclusions:Our research indicates that nuclear size, especially nuclear morphometric parameter, perimeter, provides a valuable clinicopathological index, which is useful not only in differentiating breast cancer cells from normal cells, but in differentiating molecular subtypes.

Objective:To study and verify the molecular mechanism of Duzhong Pills for osteoporosis (OP) by means of network pharmacology and molecular docking.Methods:The main chemical components of Duzhong Pills were mined by TCMSP database and the related targets were predicted. The potential targets of osteoporosis in GeneCards, DisGeNET and OMIM databases were searched and the common targets of both were obtained. The STRING platform was used for protein interaction analysis and PPI network diagram was made. The common targets were imported into the David database for enrichment analysis of GO and KEGG pathways, and molecular docking of the main components and core targets was performed. Eighteen Sprague-Dawley rats were divided into control group, model group and Duzhong Pills group according to random number table method, with 6 rats in each group. Ovariectomy was used to make osteoporosis model in model group and Duzhong Pills group. Duzhong Pills group was intragaxed with Duzhong Pills extract of 5 g/kg, and control group and model group were intragaxed with normal saline of the same volume, once a day for 8 weeks. Serum levels of TNF-α, IL-6 and IL-1β were detected by ELISA, and femur PI3K and Akt were detected by Western blot.Results:34 active components were obtained from Duzhong Pills, corresponding to 243 targets, and 1 059 targets for osteoporosis. The core targets included TNF-α, IL-6, AKT1, TP53, IL-1β and others regulated osteoporosis through PI3K-Akt and TNF pathway. The experimental results indicated that compared with model group, the levels of serum TNF-α, IL-6 and IL-1β in Duzhong Pills group decreased ( P<0.05), and the expressions of PI3K and Akt in femur decreased ( P<0.05). Conclusion:Through β-sitosterol, quercetin, kaempferol and other active components, Duzhong Pills can act on TNF, IL-6, AKT1, TP53, IL-1β and other targets, regulating PI3K-Akt signaling pathway, TNF signaling pathway and other signaling pathways to play a role in the treatment of osteoporosis.