Objective While clinical trials exploring stem cells for regenerating periodontal tissues have demonstrated positive results, there is a limited availability of systematic literature reviews on this subject. To gain a more comprehensive understanding of stem cell interventions in periodontal regeneration, this meta-analysis is undertaken to assess the beneficial effects of stem cells in human periodontal regeneration.Methods "PubMed," "PubMed Central," "Web of Science," "Embase Scopus" "Wanfang," and "CNKI," were used to extract clinical studies related to the utilization of stem cells in repairing periodontal tissue defects. This search included studies published up until October 5, 2023. The inclusion criteria required the studies to compare the efficacy of stem cell-based therapy with stem cell-free therapy for regenerating periodontal tissues. Meta-analysis was conducted using Review Manager software (version 5.4).Results This meta-analysis synthesized findings from 15 selected studies investigating the impact of stem cell interventions on periodontal tissue regeneration. The "stem cell" group displayed a substantial reduction in clinical attachment level (CAL) compared to the "control" group within 3 to 12 months post-surgery. However, no significant differences in CAL gain were found between groups. Probing pocket depth (PPD) significantly decreased in the "stem cell" group compared to the "control" group, particularly for follow-up periods exceeding 6 months, and dental stem cell treatment exhibited notable improvements. Conversely, no significant differences were observed in PPD reduction. Gingival recession (GR) significantly decreased in the "stem cell" group compared to the "control" group at 3 to 12 months post-surgery. No significant differences were observed in GR reduction between groups. No significant differences were identified in cementoenamel junction-bone distance reduction, infrabony defect reduction, or bone mineral density increase between the two groups. Furthermore, no significant changes were observed in the gingival index, plaque index, or width of keratinized gingiva.Conclusion In conclusion, while stem cell-based therapy offers promising prospects for periodontal defect treatment, there are notable limitations in the current body of research. Larger, multicenter, double-blind RCTs with robust methodologies are needed to provide more reliable evidence for stem cell-based intervention in periodontitis.

Endometriosis is a common chronic gynecological disease with endometrial cell implantation outside the uterus. Angiogenesis is a major pathophysiology in endometriosis. Our previous studies have demonstrated that the prodrug of epigallocatechin gallate (ProEGCG) exhibits superior anti-endometriotic and anti-angiogenic effects compared to epigallocatechin gallate (EGCG). However, their direct binding targets and underlying mechanisms for the differential effects remain unknown. In this study, we demonstrated that oral ProEGCG can be effective in preventing and treating endometriosis. Additionally, 1D and 2D Proteome Integral Solubility Alteration assay-based chemical proteomics identified metadherin (MTDH) and PX domain containing serine/threonine kinase-like (PXK) as novel binding targets of EGCG and ProEGCG, respectively. Computational simulation and BioLayer interferometry were used to confirm their binding affinity. Our results showed that MTDH-EGCG inhibited protein kinase B (Akt)-mediated angiogenesis, while PXK-ProEGCG inhibited epidermal growth factor (EGF)-mediated angiogenesis via the EGF/hypoxia-inducible factor (HIF-1a)/vascular endothelial growth factor (VEGF) pathway. In vitro and in vivo knockdown assays and microvascular network imaging further confirmed the involvement of these signaling pathways. Moreover, our study demonstrated that ProEGCG has superior therapeutic effects than EGCG by targeting distinct signal transduction pathways and may act as a novel antiangiogenic therapy for endometriosis.

Endometriosis is a chronic, painful disease whose etiology remains unknown. The development of novel therapies and diagnostic tools for endometriosis has been limited due in part to challenges in studying the disease. Recently, a few reports have shown that immunosuppressive cells, such as myeloid-derived suppressor cell (MDSC), may promote the progression of endometriosis. MDSCs are a heterogeneous group of myeloid cells with potent immunosuppressive and angiogenic properties. Here, in this chapter, we provide a detailed protocol to phenotype MDSC as well as to isolate and assess the functionality from the peritoneal cavity of a mouse model of surgically induced endometriosis. Importantly, the current mouse model has been widely used to study how the immune system, hormones, and environmental factors affect endometriosis as well as the effects of endometriosis on fertility and pain.

Polycystic ovary syndrome (PCOS) is a complex, heterogeneous syndrome of uncertain etiology characterized by hyperandrogenemia, hyperinsulinemia, chronic anovulation, and polycystic ovaries. Recent studies showed that the abnormalities of mitochondrial metabolism are related to PCOS. We hypothesized that mtDNA copy number is an important marker that can reflect mitochondrial function. In this study, 135 PCOS patients and 57 age-matched healthy participants were studied. Mitochondrial DNA copy number in peripheral blood and PCOS ovarian tissues, and some clinical parameters were assessed. From the single factor analysis, we can find some clinical parameters are different between PCOS and healthy women and the mitochondrial DNA copy numbers in peripheral blood in PCOS women were significantly lower than in healthy women. We also found that there was no correlation between mtDNA copy numbers in peripheral blood and in ovarian tissue. After multiple logistic regressions, we identified the occurrence of PCOS was significantly positively correlated with BMI and pulse, and negatively correlated with mitochondrial DNA copy numbers in peripheral blood. We also found the decreased mtDNA copy numbers in PCOS patients are independent of these clinical parameters. We constructed the ROC curve based on these risk factors and found if they have potential to predict the patient’s outcome. In conclusion, the changes in mtDNA copy number and some clinical parameters may provide new ideas for PCOS diagnosis. More studies are necessary for further validation of their use in PCOS diagnosis.

The mechanisms underlining pathogenesis of polycystic ovary syndrome (PCOS) remain largely unknown. Dysfunction of ovarian granulosa cells plays an important role. The present study performed the lncRNA and mRNA profiling by whole genome transcriptomic sequencing of ovary granulosa cells from women with PCOS and investigated the potential role of differentially expressed gens (DEGs) in the pathomechanism of PCOS. In total, 1,936 DEGs (30 upregulated and 1,906 downregulated mRNAs and lncRNAs) were identified in the ovary granulosa cells between control and PCOS group. Functional enrichment analysis showed that DEGs were mainly associated with cytokine-cytokine receptor interaction, neuroactive ligand-receptor interaction, and olfactory transduction. qRT-PCR validated the upregulation of DLGAP5 mRNA in ovary from PCOS group when compared to control group. Immunostaining and TUNEL assays showed that DLGAP5 protein level was increased while apoptosis was decreased in follicles of ovary in PCOS group. In vitro functional assays showed that DLGPA5 knockdown repressed viability and proliferation, but enhanced apoptosis and disrupted cell cycle in granulosa cells; while DLGAP5 overexpression had the opposite effects in granulosa cells. In conclusion, the study showed differentially expressed lncRNA and mRNA profile in the granulosa cells in ovaries of PCOS. Functional results demonstrated that DLGAP5 is a dysregulated candidate gene in the pathogenesis of PCOS, especially granulosa cell apoptosis and proliferation.

Peroxisome proliferator-activated receptor γ2 (PPARγ2) is a nuclear hormone receptor of ligand-dependent transcription factor with a key role in adipogenesis and insulin sensitization in diabetes mellitus. In this study, we investigated genetic variants in PPARγ2 promoter, its association with gestational diabetes mellitus (GDM), and its molecular regulation. PPARγ2 promoter and start codon (-2,091 to +82 bp) from 400 pregnancies with GDM and 400 gestational-age matched control pregnancies were sequenced. Association and linkage disequilibrium of the identified polymorphisms with GDM was determined. ChIP-seq, gene silencing, and dual-luciferase reporter assays were performed to confirm transcription factor binding sites and promoter activity of the variants. Transfection experiments were carried out to determine the effects of variants on gene expression and adipogenesis. Among 15 variants identified, 7 known variants were not significantly associated with the risk of GDM (odds ratio: 0.710–1.208, 95% confidence interval: 0.445–0.877 to 1.132–1.664, P > 0.05) while linkage disequilibrium was significant (D’ > 0.7, R2 > 0.9). However, T-A-A-T-G haplotype was not significantly associated with GDM (χ2 = 2.461, P = 0.117). Five rare variants and 3 novel variants (rs948820149, rs1553638909, and rs1553638903) were only found in GDM. Transcription factor glucocorticoid receptor β (GRβ) bound to -807A/C (rs948820149) and knockdown of GRβ suppressed PPARγ2 promoter activity. This mutation significantly down-regulated PPARγ2 expression and alleviated adipogenesis. In conclusion, a novel -807A/C in PPARγ2 promoter was identified in Chinese women with GDM and the mutation affected GRβ binding and transcription of PPARγ2 for adipogenesis.

Abstract High risk Human Papillomavirus (HPVs) is a very important etiological agent in cervical cancer. HPV types have been subdivided into low-risk types and high-risk types, which are frequently associated with invasive cervical cancer. HPV16 and HPV18 count for 70% of cervical cancer. The E6, E7 genes in the early region of HPV genome are proved to have a significant impact on progression of cervical cancer. miRNA is a kind of short non-coding RNA which regulates the gene expression on post-transcriptional and translational level, and it has been proved to have a significant impact on tumor development. Our group previously investigated the correlation of miRNAs and HPV oncogenes, and identified several differentially expressed miRNAs in cervical cancer cell lines and clinical specimens. The expression level of miR-145, miR-195 and miR-142-3p were down-regulated in cervical cancer patients. However, whether the HPV oncogenes regulate miRNAs is still unknown. In order to determine the relationship between HPV oncogenes and miRNAs. We did several experiments. We found that up/down-regulate HPV 16/18 E6 and E7 caused an decrease/increase of BRCA1, miR-145 and miR-195, and BRCA1 could increase the expression of miR-145 and miR-195 in different cell lines transfected with HPV 16/18 E6 and E7 by using qRT-PCR and Western-blot. We also found the binding activity of E2F1 and MYC to the promoter region of BRCA1, which validated by ChIP-PCR and Luciferase assay. These pathways were also validated in tumor xenograft nude mice. We also tried to find the target genes of miR-145 and miR-195, and investigated how they affect tumorigenesis in cervical cancer. These findings provided a solid foundation that HPV 16/18 E6 and E7 regulated miR-145 and miR-195 via BRCA1 in cervical cancer. It can be an important pathway for the pathogenesis of cervical cancer induced by high risk HPV. These findings could provide new ways in diagnosis and treatment for cervical cancer. They also give a new insight to investigate molecular pathways for HPV-associated cancers. We believe that further investigation will be done for the mechanisms in the future. Citation Format: Yi Song, Yan Deng, Tao Tang, Chi Chiu Wang. The mechanism of HPV oncogenes in the regulation of upstream microRNA pathway in cervical cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2531.

Background Previous studies have shown a major green tea polyphenol (−)-epigallocatechin-3-gallate ((−)-EGCG) as a powerful anti-cancer agent. However, its poor bioavailability and requirement of a high dosage to manifest activity have restricted its clinical application. Recently, our team synthesized a peracetate-protected derivative of EGCG, which can act as a prodrug of (−)-EGCG (ProEGCG) with enhanced stability and improved bioavailability in vitro and in vivo. Herein, we tested the therapeutic efficacy of this novel ProEGCG, in comparison to EGCG, toward human endometrial cancer (EC). Methods In this study, the effects of ProEGCG and EGCG treatments on cell growth, cell survival and modulation of intracellular signaling pathways in RL95–2 and AN3 CA EC cells were compared. The antiproliferative effect was evaluated by cell viability assay. Apoptosis was measured by annexin/propidium iodide staining. Expression of mitogen-activated protein kinases, markers of proliferation and apoptosis were measured by immunoblot analysis. In addition, the effects of ProEGCG and EGCG on tumor growth, vessel formation and gene expression profiles on xenograft models of the EC cells were investigated. Results We found that treatment with ProEGCG, but not EGCG, inhibited, in a time- and dose-dependent manner, the proliferation and increased apoptosis of EC cells. Treatment with low-dose ProEGCG significantly enhanced phosphorylation of JNK and p38 MAPK and inhibited phosphorylation of Akt and ERK which are critical mediators of apoptosis. ProEGCG, but not EGCG, elicited a significant decrease in the growth of the EC xenografts, promoted apoptotic activity of tumour cells in the EC xenografts, and decreased microvessel formation, by differentially suppressing anti-apoptotic molecules, NOD1 and NAIP. Notably, no obvious adverse effects were detected. Conclusions Taken together, ProEGCG at a low dose exhibited anticancer activity in EC cells through its anti-proliferative, pro-apoptotic and anti-tumor actions on endometrial cancer in vitro and in vivo. In contrast, a low dose of EGCG did not bring about similar effects. Importantly, our data demonstrated the efficacy and safety of ProEGCG which manifests the potential of a novel anticancer agent for the management of endometrial cancer.

High risk Human Papillomavirus (HPVs) infection is the etiological agent in cervical cancer. HPV types have been subdivided into low-risk types, which are found mainly in genital warts, and high-risk types, which are frequently associated with invasive cervical cancer. HPV16 and HPV18 count for 70% of cervical cancer. The E6, E7 genes in the early region of HPV genome are proved to have a significant impact on progression of cervical cancer. miRNA is a kind of short (about 22nt) non-coding RNA which regulates the gene expression on post-transcriptional and translational level, and it has been proved to have a significant impact on tumor development. Our group previously investigated the correlation of miRNAs and HPV oncogenes, and identified several differentially expressed miRNAs in cervical cancer cell lines (HeLa, SiHa, C33A, ME180, Caski, CC2 and CC3) as well as clinical specimens, including 2 up-regulated and 9 down-regulated. We further investigate the direct relationship between the oncogenes and miRNAs. C33A cells were transfected with single E6/E7 vector or E6/E7 vector + E6/E7 siRNA, and the expression of miRNAs were tested through qRT-PCR. The result of the relative expression level of miRNAs shows some consistency. The HPV oncogenes could down-regulate the expression of miR-145, miR-195 and miR-142-3p. We also have examined the oncogenetic effects of miR-145, miR-195 and miR-142-3p by MTT assay, which shows they all reduce the cell viability. Further investigation will be done for the mechanisms. The work could provide new insight in diagnosis and treatment for cervical cancer.

Accumulating experimental evidence has shown that the aberrant expression of microRNAs (miRNAs) is involved in the development and progression of human cervical cancer. Previously, we identified miR-182 as an oncomiRNA in cervical cancer. However, the mechanism by which miR-182 is regulated and the interaction between human papillomavirus (HPV) and miR-182 in cervical cancer development remains unknown. In the present study, we explored the link between HPV E7 and miR-182 and verified that high-risk HPV E7 upregulated miR-182 expression through TGF-β/Smad4 signaling pathway in cervical cancer. By contrast, low-risk HPV E7 did not affect the expression of TGF-β and miR-182. Mechanistically, as high-risk HPV E7 bound to pRb, E2F was released from the complex and bound to the TGF-β promoter region, resulting in TGF-β overexpression. Furthermore, the Smad4 signaling pathway was activated upon TGF-β overexpression, which led to an interaction between Smad4 and the miR-182 promoter region, subsequently inducing the upregulation of miR-182 in both cervical cancer cells and the surrounding normal cells. In conclusion, this newly identified high-risk HPV E7/TGF-β/miR-182 regulatory network might inform the development of specific therapeutic strategies for cervical cancer.

OBJECTIVE:This study was conducted to explore chromosomal copy number variations (CNV) and transcript expression and to examine pathways in cervical pathogenesis using genome-wide high resolution microarrays.METHODS:Genome-wide chromosomal CNVs were investigated in 6 cervical cancer cell lines by Human Genome CGH Microarray Kit (4x44K). Gene expression profiles in cervical cancer cell lines, primary cervical carcinoma and normal cervical epithelium tissues were also studied using the Whole Human Genome Microarray Kit (4x44K).RESULTS:Fifty common chromosomal CNVs were identified in the cervical cancer cell lines. Correlation analysis revealed that gene up-regulation or down-regulation is significantly correlated with genomic amplification (P=0.009) or deletion (P=0.006) events. Expression profiles were identified through cluster analysis. Gene annotation analysis pinpointed cell cycle pathways was significantly (P=1.15E-08) affected in cervical cancer. Common CNVs were associated with cervical cancer.CONCLUSION:Chromosomal CNVs may contribute to their transcript expression in cervical cancer.