Zishen Qingre Lishi Huayu recipe promotes proliferation and inhibits apoptosis of GCs of PCOS via KLF4-C/EBPß pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Zishen Qingre Lishi Huayu recipe (ZQLHR) is a herbal recipe created on the basis on the theory of traditional Chinese medicine and clinical practice, and is mainly used in the treatment of polycystic ovary syndrome (PCOS). However, the underlying mechanism for this fact has not been clearly elucidated. AIM OF THE STUDY: To verify whether ZQLHR regulates granulosa cells (GCs) proliferation and apoptosis through the Krüppel-like factor 4 (KLF4) - CCATT enhancer-binding proteinß (C/EBPß) pathway, and to provide in vitro molecular mechanism supporting for the effects of ZQLHR to enhance follicular development and treat patients with PCOS. MATERIALS AND METHODS: Based on previous experiments, we performed the following experiments. Firstly, we treated KGN cells (a steroidogenic human granulosa-like tumor cell line) for 48 h using different concentrations of ZQLHR in order to observe apoptosis in each group. Secondly, the mRNA and protein expression levels of KLF4 and C/EBPß in KGN cells after administrated with ZQLHR were examined by quantitative real-time PCR(q-PCR) and Western blot assay. Thirdly, after knocking down KLF4 and C/EBPß using siRNAs, the relationship between KLF4 and C/EBPß in KGN cells was detected. Further, cell counting kit-8 assay, colony formation assay and flow cytometry were used to verify whether ZQLHR promotes proliferation and facilitates apoptosis in KGN cells through the KLF4-C/EBPß pathway. Finally, q-PCR and Western blot were used to test whether ZQLHR mediated proliferation and apoptosis-related factors such as B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (BAX), proliferating cell nuclear antigen (PCNA) and cleaved caspase-3 to affect the proliferation and apoptosis of KGN cells through the KLF4-C/EBPß pathway. CONCLUSIONS: ZQLHR, containing 0.2% by volume, administered to KGN cells resulted in the lowest rate of apoptosis. The expression levels of KLF4 and C/EBPß were increased in KGN cells following ZQLHR treatment. Additionally, ZQLHR promoted proliferation and inhibited apoptosis of KGN cells by modulating proliferation and apoptosis-related factors via the KLF4-C/EBPß pathway. Furthermore, we confirmed that KLF4 and C/EBPß regulate each other in KGN cells. These findings indicate that ZQLHR enhances the proliferation of GCs and suppresses their apoptosis, which constitutes a therapeutic mechanism for treating patients with PCOS.

Temporal and spatial features of the palace building space of Qinghai's Kumbum Monastery and its evolution.

Kumbum Monastery is the centre of Tibetan Buddhism and religious culture in Qinghai. Its palace buildings are the typical examples of Tibetan Buddhist monastery buildings and living fossils of Tibetan social history. This study selected 12 palace buildings of Kumbum Monastery as the study objects, used typological approaches and numerical method to analyse their spatial features, classified these features into four types (the ring road surrounding Dugang-style palace space, single-sided eaves of the Duguang-style palace space, three-stage palace space with cloisters connecting to Dugang, and other variants) according to their spatial structures, and discussed the temporal evolution of the spatial features from spatial and temporal perspectives to obtain the development process for the palace buildings of Kumbum Monastery. The analysis showed that: 1) Because the Han people migrated to Qinghai, the architectural space of the monastery followed the practices of Tibet in the same period and began to adopt the practices of the Han people under the effect of religious indoctrination and sociopolitical influence of the Ming dynasty. 2) Due to the influence of religious development facilitated by the political environment in the Qing dynasty, all the palace buildings of Kumbum Monastery adopted the practice of monasteries in Lhasa. After being implicated by the political rebellion, the monastery initiated to add the spatial layout elements of the buildings in Qinghai. Therefore, the monastery was obtaining the cues from the Han culture. 3) The dominance of religious significance over the spatial designing of the palace buildings of Kumbum Monastery gradually shifted to the political dominance. This paper revealed the spatial and temporal features and evolution of the palace building space, explored the generation process of the palace buildings space of Tibetan Buddhist monasteries in Qinghai, and provided a reference for the static conservation and dynamic development of Tibetan Buddhist monastery buildings in Qinghai.

Protection and Planning of Historic Districts Based on Internet of Things Perception.

With the development of the Internet of Things era, the Internet of Things technology has been gradually accepted by people and gradually entered the category of block management, but the specific practical application scope is not wide enough. At present, the community management of residents in blocks generally adopts closed management, and there are some problems such as insufficient scope of property management, weak strength, and serious waste of human resources. Incomplete development of the hidden value is of cultural tourism blocks. At the same time, there are still some problems in the traffic management of blocks, such as incomplete monitoring scope and weak supervision. Therefore, this study proposes a bilevel programming model to solve the optimization scheme of block road network, adopts the complete information game model to analyze the imbalance between property and residents, and adopts the AHP fuzzy evaluation method to detect the current life happiness coefficient of urban residents and collect residents' suggestions for the future planning of historical blocks. Because the perception layer of the Internet of Things has the advantages of rapidity, accuracy, and unmanned operation, it can contribute a practical force to the protection and planning of historical blocks quickly and at low cost.

Genome-Wide Analysis of Coding and Long Non-Coding RNAs Involved in Cuticular Wax Biosynthesis in Cabbage (Brassica oleracea L. var. capitata).

Cuticular wax is a mixture of very long chain fatty acids (VLCFAs) and their derivatives, which determines vital roles for plant growth. In cabbage, the cuticular wax content of leaf blades is an important trait influencing morphological features of the head. Understanding the molecular basis of cuticular wax biosynthesis can help breeders develop high quality cabbage varieties. Here, we characterize a cabbage non-wax glossy (nwgl) plant, which exhibits glossy green phenotype. Cryo-scanning electron microscope analysis showed abnormal wax crystals on the leaf surfaces of nwgl plants. Cuticular wax composition analyzed by GC-MS displayed severely decreased in total wax loads, and individual wax components in nwgl leaves. We delimited the NWGL locus into a 99-kb interval between the at004 marker and the end of chromosome C08 through fine mapping. By high-throughput RNA sequencing, we identified 1247 differentially expressed genes (DEGs) and 148 differentially expressed lncRNAs in nwgl leaves relative to the wild-type. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that the DEGs and cis-regulated target genes for differentially expressed lncRNAs were significantly enriched in wax and lipid biosynthetic or metabolic processes. Our results provide the novel foundation to explore the complex molecular basis of cuticular wax biosynthesis.

[Optimization of SRAP & ISSR technology and its application in the identification of seeds of Brassica oleracea L].

In this study, the molecular marker technology of SRAP and ISSR were applied in rapid identification of seeds from eight species of Brassica oleracea L. Firstly, using the genomic DNA of cabbage as template, SRAP and ISSR reaction systems were optimized through testing every factor, respectively, that affects PCR amplification. Then, using the optimized reaction systems, 30 SRAP primer pairs and 15 ISSR primers were applied to amplify genomic DNA of cabbage, savoy, purple cabbage, borecole, cauliflower, broccoli, Brussels sprouts, and kohlrabi The results showed that high polymorphisms were exhibited among the eight species of Brassica oleracea L. by SRAP primer pairs of M3-E5 and M4-E5, as well as ISSR primers of 844 and 888, especially primer 844 which can identify all eight materials efficiently.