The novel HLA-DQB1*03:03:29 allele, identified by sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-DQB1*03:03:29 differs from HLA-DQB1*03:03:02:01 by one nucleotide in exon 2.

[A correlation study of moxibustion time and moxibustion effect of long snake moxibustion in intervention of recurrent exopathogenic diseases in patients with yang-deficiency constitution].

OBJECTIVE: To explore the time-effect relationship of long snake moxibustion in intervening recurrent exopathogenic diseases of patients with yang-deficiency constitution in different moxibustion periods and provide a scientific basis for the selection of long snake moxibustion in preventing and treating recurrent exopathogenic diseases of patients with yang-deficiency constitution. METHODS: Ninety patients with yang-deficiency constitution who met the inclusion criteria of recurrent exopathogenic diseases were randomly divided into a 30 min group, a 60 min group, and a 90 min group, with 30 cases in each group. Long snake moxibustion was applied once a week from Dazhui (GV14) to Yaoshu (GV2) for different periods (30, 60, and 90 min), 12 times (12 weeks) in total. The scores of yang-deficiency constitution quality scale and Fatigue Severity Scale (FSS) before treatment, after treatment, and six months after treatment, as well as attack times of exopathogenic diseases within one year before treatment and after treatment in the three groups, were observed and recorded. RESULTS: After treatment and 6 months after treatment, the yang-deficiency quality scale scores and FSS scores of the three groups were lower than those before treatment (P<0.01), which decreased in the sequence of the 30 min group, the 90 min group, and the 60 min group (P<0.05). Within one year after treatment, the attack times of exopathogenic diseases in the three groups was lower than that within one year before treatment (P<0.01), which decreased in the sequence of the 30 min group, the 90 min group, and the 60 min group (P<0.05). CONCLUSION: The optimal moxibustion time of long snake moxibustion on the recurrent exopathogenic diseases of patients with yang-deficiency constitution is 60 min.

[Diagnostic value of physical examination tests for supraspinatus tendon tears].

OBJECTIVE: To investigate the diagnostic value of 6 conventional physical examination tests for the diagnosis of supraspinatus tendon tears, and how well they could tell the difference between partial-and full-thickness tears. METHODS: A total of 91 patients with different shoulder symptoms who received shoulder arthroscopic procedure were enrolled in the study from June 2017 to September 2020. The intraoperative findings were compared with the results of the preoperative physical examination of 6 clinical tests, including the Hug-up test, the Jobe test, the 0°abduction test, the drop arm test, the Neer test, and the Hawkins test, to determine the sensitivity, specificity, positive and negative predictive value, accuracy, positive and negative likelihood ratio of each test. RESULTS: By arthroscopy, a total of 44 full-thickness tears, 34 partial-thickness tears, and 13 intact supraspinatus tendons were found in all 91 cases. The Hug-up and the Jobe tests significantly correlated with the intraoperative findings. The sensitivity of the Hug-up test, the Jobe test, the 0° abduction test, the drop arm test, the Neer test, and the Hawkins test was 0.90, 0.79, 0.64, 0.42, 0.49, 0.24 respectively;the specificity was 0.61, 0.69, 0.54, 0.38, 0.31, 0.77;the positive predictive value was 0.93, 0.94, 0.89, 0.80, 0.81, 0.86;the negative predictive value was 0.50, 0.36, 0.20, 0.10, 0.09, 0.14;the accuracy was 0.86, 0.78, 0.63, 0.42, 0.46, 0.32;the positive likelihood ratio was 2.30, 2.58, 1.39, 0.69, 0.71, 1.06;and the negative likelihood ratio was 0.16, 0.30, 0.67, 1.50, 1.65, 0.98. CONCLUSION: The Jobe test and the Hug-up test are both effective at accurately diagnosing supraspinatus tendon tears, the Hug-up test detects supraspinatus tears with a high sensitivity, and similar specificity. The tests assessed in this study are not capable of distinguish between partial-and full thickness supraspinatus tendon tears.

Diagnostic value of physical examination tests for supraspinatus tendon tears / 中国骨伤

OBJECTIVE@#To investigate the diagnostic value of 6 conventional physical examination tests for the diagnosis of supraspinatus tendon tears, and how well they could tell the difference between partial-and full-thickness tears.@*METHODS@#A total of 91 patients with different shoulder symptoms who received shoulder arthroscopic procedure were enrolled in the study from June 2017 to September 2020. The intraoperative findings were compared with the results of the preoperative physical examination of 6 clinical tests, including the Hug-up test, the Jobe test, the 0°abduction test, the drop arm test, the Neer test, and the Hawkins test, to determine the sensitivity, specificity, positive and negative predictive value, accuracy, positive and negative likelihood ratio of each test.@*RESULTS@#By arthroscopy, a total of 44 full-thickness tears, 34 partial-thickness tears, and 13 intact supraspinatus tendons were found in all 91 cases. The Hug-up and the Jobe tests significantly correlated with the intraoperative findings. The sensitivity of the Hug-up test, the Jobe test, the 0° abduction test, the drop arm test, the Neer test, and the Hawkins test was 0.90, 0.79, 0.64, 0.42, 0.49, 0.24 respectively;the specificity was 0.61, 0.69, 0.54, 0.38, 0.31, 0.77;the positive predictive value was 0.93, 0.94, 0.89, 0.80, 0.81, 0.86;the negative predictive value was 0.50, 0.36, 0.20, 0.10, 0.09, 0.14;the accuracy was 0.86, 0.78, 0.63, 0.42, 0.46, 0.32;the positive likelihood ratio was 2.30, 2.58, 1.39, 0.69, 0.71, 1.06;and the negative likelihood ratio was 0.16, 0.30, 0.67, 1.50, 1.65, 0.98.@*CONCLUSION@#The Jobe test and the Hug-up test are both effective at accurately diagnosing supraspinatus tendon tears, the Hug-up test detects supraspinatus tears with a high sensitivity, and similar specificity. The tests assessed in this study are not capable of distinguish between partial-and full thickness supraspinatus tendon tears.

[Genotype Analysis of Pregnant Women with α- and ß- Thalassemia in Fuzhou Area of Fujian Province in China].

OBJECTIVE: To analyze the genotype of pregnant women with α- and ß- thalassemia in Fuzhou area of Fujian province in China. METHODS: Blood routine examination and hemoglobin electrophoresis were performed for pregnant women, and positive samples were examined by gap polymerase chain reaction and reverse dot blot hybridization. RESULTS: 412 cases were diagnosed as α-thalassemia (63.9%); 201 cases were diagnosed as ß-thalassemia (31.2%); 32 cases were diagnosed as α and ß-composite thalassemia. There were 12 genotypes in α-thalassemia, whose major genotypes were --SEA/αα, α3.7/αα, -α4.2/αα and αQSα/αα, with carrying rate of 64.32%, 20.14%, 7.77% and 1.94%, respectively. There were 10 genotypes in ß- thalassemia, whose major genotypes were CD41-42/N, CD17/N, IVS-II-654/N and -28/N, with carrying rate of 30.84%, 27.86%, 15.92% and 10.45%, respectively. There were 9 genotypes in α and ß-composite thalassemia, whose major genotypes were --SEA/αα composited CD41-42/N, -α3.7/αα composited CD41-42/N, --SEA/αα composited CD17/N, with carrying rate of 18.75%, 15.62%, 15.62% respectively. CONCLUSION: The major genotypes of pregnant women with α- and ß- thalassemia in Fuzhou area of Fujian province in China are --SEA/αα, α3.7/αα, CD41-42/N and CD17/N. Thalassemia screening and prenatal gene diagnosis should be strengthened in Fuzhou area of Fujian province in China.

Genotype Analysis of Pregnant Women with α- and β- Thalassemia in Fuzhou Area of Fujian Province in China / 中国实验血液学杂志

OBJECTIVE@#To analyze the genotype of pregnant women with α- and β- thalassemia in Fuzhou area of Fujian province in China.@*METHODS@#Blood routine examination and hemoglobin electrophoresis were performed for pregnant women, and positive samples were examined by gap polymerase chain reaction and reverse dot blot hybridization.@*RESULTS@#412 cases were diagnosed as α-thalassemia (63.9%); 201 cases were diagnosed as β-thalassemia (31.2%); 32 cases were diagnosed as α and β-composite thalassemia. There were 12 genotypes in α-thalassemia, whose major genotypes were --/αα, α/αα, -α/αα and αα/αα, with carrying rate of 64.32%, 20.14%, 7.77% and 1.94%, respectively. There were 10 genotypes in β- thalassemia, whose major genotypes were CD41-42/N, CD17/N, IVS-II-654/N and -28/N, with carrying rate of 30.84%, 27.86%, 15.92% and 10.45%, respectively. There were 9 genotypes in α and β-composite thalassemia, whose major genotypes were --/αα composited CD41-42/N, -α/αα composited CD41-42/N, --/αα composited CD17/N, with carrying rate of 18.75%, 15.62%, 15.62% respectively.@*CONCLUSION@#The major genotypes of pregnant women with α- and β- thalassemia in Fuzhou area of Fujian province in China are --/αα, α/αα, CD41-42/N and CD17/N. Thalassemia screening and prenatal gene diagnosis should be strengthened in Fuzhou area of Fujian province in China.

Evidence for and against Liquid-Liquid Phase Separation in the Nucleus.

Enclosed by two membranes, the nucleus itself is comprised of various membraneless compartments, including nuclear bodies and chromatin domains. These compartments play an important though still poorly understood role in gene regulation. Significant progress has been made in characterizing the dynamic behavior of nuclear compartments and liquid-liquid phase separation (LLPS) has emerged as a prominent mechanism governing their assembly. However, recent work reveals that certain nuclear structures violate key predictions of LLPS, suggesting that alternative mechanisms likely contribute to nuclear organization. Here, we review the evidence for and against LLPS for several nuclear compartments and discuss experimental strategies to identify the mechanism(s) underlying their assembly. We propose that LLPS, together with multiple modes of protein-nucleic acid binding, drive spatiotemporal organization of the nucleus and facilitate functional diversity among nuclear compartments.

Interleukin-17 plays a role in pulp inflammation partly by WNT5A protein induction.

OBJECTIVE: Our study aimed to investigate the role of interleukin (IL)-17 in dental pulp inflammation and the relationship between WNT5A and IL-17. METHODS: Immunohistochemical staining was used to detect the expression of tumor necrosis factor-α (TNF-α), WNT5A and IL-17 in pulp tissues. Anti-IL-17 neutralizing antibody was used in rat pulpitis model and to study the role of IL-17 in pulpitis. TNF-α, WNT5A or IL-17 recombinant protein were used to treat human dental pulp cells. RT-PCR, Western blot, and Enzyme linked immunosorbent assay were used to detect the expression of mRNA and protein. Transwell assay was used to measure the migration of THP-1 cells, which is a human monocytic cell line. RESULTS: IL-17 and WNT5A are co-expressed in TNF-α high-expressed region in human and rat pulpitis tissue. IL-17 mainly contributes to its positive regulatory role in inflammation through up regulate cytokines and mediated macrophages migration. Anti-IL-17 neutralizing antibody can suppress the inflammatory cell infiltration and TNF-α expression in dental pulpitis. TNF-α promotes the expression of IL-17 partly through WNT5A and WNT5A regulates IL-17 expression by mitogen-activated protein kinase (MAPK)-(P38 and ERK) pathway. CONCLUSIONS: IL-17 acts as an inflammatory mediator in dental pulp inflammation. The expression of IL-17 can be partially regulated by WNT5A.

Antibacterial activities of a series of novel 5-O-(4', 6'-O-dimodified)-mycaminose 14-membered ketolides.

A series of novel 5-O-(4',6'-O-dimodified)-mycaminose 14-membered ketolides were assessed for their in vitro antibacterial activities against a panel of sensitive and resistant pathogens. Compound 1 and compound 2, two ester analogs, showed the best antibacterial activities against several macrolide-sensitive and macrolide-resistant strains. These results indicated that introducing ester to 6-OH and a small volume ether substituent to the 4-OH of mycaminose could improve the antibacterial activities of ketolides.

A Bioinformatic Profile of Gene Expression of Colorectal Carcinoma Derived Organoids.

Colorectal carcinoma is one of the common cancers in human. It has been intensely debated whether the in vitro cancer cell lines are closely enough for recapitulating the original tumor in understanding the molecular characteristic of CRC. Organoid as a new in vitro 3D culture system has sprang out in CRC study for the capability in reviving the original tissue. The aim of this study is to profile the gene expression of CRC organoid. The gene expression GSE64392 was from GEO database contained 20-patients-derived 37 organoid samples, including 22 colorectal tumor organoid samples and 15 paired healthy samples. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were applied for classifying differentially expressed genes (DEGs). Protein interaction among DEGs was analyzed by Search Tool for the Retrieval of Interacting Genes (STRING) and Cytoscape software. In total, 853 gene sequences were identified. GO analysis revealed that DEGs were extensively involved in various biological process (BP), like proliferation, cell cycle, and biosynthesis. KEEG pathway analysis showed that WNT, MAPK, TGF-ß, SHH, ECM-receptor interaction, and FGF pathways were altered. DEGs which were identified with protein interactions were major response for extracellular matrix organization and the GPCR pathway. In conclusion, our study profiled the DEGs in CRC organoids and promotes our understanding of the CRC organoids as a new model for colorectal cancer research.

EZH2 promotes DNA replication by stabilizing interaction of POLδ and PCNA via methylation-mediated PCNA trimerization.

BACKGROUND: Proliferating cell nuclear antigen (PCNA), a ring-shaped homotrimer complex, promotes DNA replication via binding to DNA polymerase. Trimerized PCNA is critical for DNA replication. Enhancer of zeste homologue 2 (EZH2), which primarily acts as a histone methyltransferase, is essential for proliferation. However, how EZH2 promotes proliferation by controlling DNA replication through PCNA remains elusive. RESULTS: Here, we showed that low EZH2 levels repressed the proliferation of human dental pulp cells (hDPCs). The EZH2 protein level was dramatically upregulated in hDPCs at S phase in the absence of H3K27 trimethylation. Molecularly, EZH2 interacted with PCNA via the PIP box and dimethylated PCNA at lysine 110. Dimethylation of PCNA is essential for stabilization of the PCNA trimer and the binding of DNA polymerase δ to PCNA. CONCLUSIONS: Our data reveal the direct interaction between PCNA and EZH2 and a novel mechanism by which EZH2 orchestrates genome duplication.

Design, Synthesis and Antibacterial Evaluation of 1-[(1R,2S)-2-Fluorocyclopropyl]ciprofloxacin-1,2,4-triazole-5(4H)-thione Hybrids.

A new class of 1-[(1R,2S)-2-fluorocyclopropyl]ciprofloxacin (CPFX)-1,2,4-triazole-5(4H)-thione hybrids 6a - 6o was designed, synthesized and evaluated for their in vitro antibacterial activities against a panel of clinically important drug-sensitive and drug-resistant Gram-positive and Gram-negative pathogens. Our results revealed that all hybrids 6a - 6o had great potency against the tested strains, especially Gram-negative pathogens. The synthesized hybrids were more potent than the parent 1-[(1R,2S)-2-fluorocyclopropyl]CPFX (1) and comparable to CPFX and levofloxacin against the majority of the tested pathogens, worth to be further investigated.

Inhibition of SOX9 Promotes Inflammatory and Immune Responses of Dental Pulp.

INTRODUCTION: The process of pulpitis is characterized by extracellular matrix imbalance and inflammatory cell infiltration. As an essential transcription factor, sex-determining region Y-box 9 (SOX9) is significantly inhibited by tumor necrosis factor alpha in inflammatory joint diseases. The aim of this study was to explore the role of SOX9 in extracellular matrix balance, cytokine expression, and the immune response in dental pulp. METHODS: The expression of SOX9 in normal and inflamed pulp tissue/human dental pulp cells (HDPCs) was detected by immunohistochemistry, Western blot, and quantitative polymerase chain reaction (qPCR). SOX9 small interfering RNA was used to knock down SOX9 expression of dental cells in vitro; extracellular matrix imbalance was analyzed by qPCR, Western blot, and gelatin/collagen zymography, and the secretion of cytokines was scanned by antibody arrays. The immune response of THP-1 was investigated by cell migration assay, cell attachment assay, phagocytosis assay, and enzyme-linked immunosorbent assay. The interaction of SOX9 with target genes was explored by chromatin immunoprecipitation (ChIP). RESULTS: SOX9 was strongly expressed in normal dental pulp tissue and HDPCs and reduced in inflamed pulp. SOX9 knockdown could inhibit the production of type I collagen, stimulate the enzymatic activities of MMP2 and MMP13, and regulate the production of interleukin (IL) 8 of HDPCs. SOX9 knockdown also effectively suppressed the differentiation and functional activities of THP-1. ChIP showed that the binding of the SOX9 protein with matrix metalloproteinase (MMP)-1, MMP-13, and IL-8 gene promoters was reduced after being treated with recombinant human tumor necrosis factor alpha. CONCLUSIONS: SOX9 was inhibited in inflamed dental pulp and may participate in the regulation of extracellular matrix balance, the inflammatory process, and the immune response.

EZH2 regulates dental pulp inflammation by direct effect on inflammatory factors.

OBJECTIVE: Pulpitis is a multi-factorial disease that could be caused by complex interactions between genetics, epigenetics and environmental factors. We aimed to evaluate the role of Enhancer of Zeste Homolog 2 (EZH2) in the inflammatory response of human dental pulp cells (HDPCs) and dental pulp tissues. METHODS: The expressions of inflammatory cytokines in HDPCs treated by EZH2 complex or EZH2 siRNA with or without rhTNF-α were examined by quantitative real-time polymerase chain reaction (q-PCR). The levels of secreted inflammatory cytokines including IL-6, IL-8, IL-15, CCL2 and CXCL12 in culture supernatants were measured by Luminex assay. In rat pulpitis model, the effects of EZH2 on dental pulp tissues were verified by histology. We invested the mechanisms of the effect of EZH2 on the inflammatory factors by ChIP assay. RESULTS: EZH2 down-regulation inhibited the expression of inflammatory factors, including IL-6, IL-8, IL-15, CCL2 and CXCL12 in HDPCs. EZH2 complex promoted the expression and secretion of these inflammatory factors in HDPCs, while EZH2 silencing could attenuate the promotion of inflammatory factors that were induced by rhTNF-α. In pulpitis models of rats, EZH2 down-regulation inhibited the inflammatory process of dental pulp while EZH2 complex showed no significant facilitation of pulpal inflammation. In addition, EZH2 could bind on the promoters of IL-6, IL-8 and CCL2, but not IL-15 and CXCL12, to affect the transcription of these proinflammatory cytokines. CONCLUSIONS: In HDPCs, EZH2 could induce inflammation, while EZH2 down-regulation could attenuate the inflammatory responses. EZH2 plays an important role in this inflammatory process of dental pulp.

[Effect of Complicated Hemophagocytic Syndrome on Clinical Prognosis of Patients with Non-Hodgkin's Lymphoma and Analysis of Its Affecting Factors].

OBJECTIVE: To investigate the effect of complicatal hemophagocytic syndrome on clinical prognosis of patients with non-Hodgkin's lymphoma (NHL) and analyze its factors affecting prognosis. METHODS: Ninety cases of NHL were selected and divided into 2 groups: 61 cases of NHL without hemophagocytic syndrome as group A and 29 cases of NHL with hemophagocytic syndrame as group B. The survival analysis of Kaplan-Meter method and the Cox regression model were used for univariate and multivariate analyses of related factors. RESULTS: The patients in group B were more likely to start with fever, moreover, the hemophagocytes could be found in bone marrow samples of 89.66% (26/29) patients; the levels of total bilirubin, triglycerides, serum ferritin, serum soluble CD25, DNA copies of epstein-barr virus (EBV) and lactate dehydrogenase level in the group B were significantly higher than those in the group A(P<0.05). And the patients in group B had worse physical state, later disease stage, worse disease status and lower overall prognosis as compared with patients in the group A. The complicased hemophagocytic syndrome, incomplete improvemant of deseases state after treatment and EBV infection were the independent risk factors for the poor prognosis of patients with NHL. CONCLUSION: The complicated hemophagocytic syndrome can increase the severity of NHL, there fore significantly influences the clinical prognosis of patients, while the complicated hemophagocytic syndrome, poor therapatic efficacy for patients and EBV infection are independent risk factors affecting the prognosis of NHL patients.

Synthesis and antibacterial activities of some novel 17, 18-unsaturated carbonyl compounds derivated from josamycin.

Some novel josamycin derivatives bearing an arylalkyl-type side chain were designed and synthesized. By HWE or Wittig reaction, 16-aldehyde group of josamycin analogs were converted into unsaturated carbonyl compounds. They were evaluated for their in vitro antibacterial activities against a panel of respiratory pathogens. 8b and 8e exhibited comparable activities against a panel of respiratory pathogens, especially to resistant ones in the series of desmycarosyl josamycin analogs. Among of all the target molecules, 21 showed the best antibacterial activities.

Universal quantum gates for hybrid system assisted by atomic ensembles embedded in double-sided optical cavities.

We propose deterministic schemes for controlled-NOT (CNOT), Toffoli, and Fredkin gates between flying photon qubits and the collective spin wave (magnon) of an atomic ensemble inside double-sided optical microcavities. All the gates can be accomplished with 100% success probability in principle and no additional qubit is required. Atomic ensemble is employed so that light-matter coupling is remarkably improved by collective enhancement. We qualified the performance of the gates and the results show that they can be faithfully constituted with current experimental techniques.

Effect of Complicated Hemophagocytic Syndrome on Clinical Prognosis of Patients with Non-Hodgkin's Lymphoma and Analysis of Its Affecting Factors / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of complicatal hemophagocytic syndrome on clinical prognosis of patients with non-Hodgkin's lymphoma (NHL) and analyze its factors affecting prognosis.</p><p><b>METHODS</b>Ninety cases of NHL were selected and divided into 2 groups: 61 cases of NHL without hemophagocytic syndrome as group A and 29 cases of NHL with hemophagocytic syndrame as group B. The survival analysis of Kaplan-Meter method and the Cox regression model were used for univariate and multivariate analyses of related factors.</p><p><b>RESULTS</b>The patients in group B were more likely to start with fever, moreover, the hemophagocytes could be found in bone marrow samples of 89.66% (26/29) patients; the levels of total bilirubin, triglycerides, serum ferritin, serum soluble CD25, DNA copies of epstein-barr virus (EBV) and lactate dehydrogenase level in the group B were significantly higher than those in the group A(P<0.05). And the patients in group B had worse physical state, later disease stage, worse disease status and lower overall prognosis as compared with patients in the group A. The complicased hemophagocytic syndrome, incomplete improvemant of deseases state after treatment and EBV infection were the independent risk factors for the poor prognosis of patients with NHL.</p><p><b>CONCLUSION</b>The complicated hemophagocytic syndrome can increase the severity of NHL, there fore significantly influences the clinical prognosis of patients, while the complicated hemophagocytic syndrome, poor therapatic efficacy for patients and EBV infection are independent risk factors affecting the prognosis of NHL patients.</p>

[Inhibitory effect of bone marrow mesenchymal stem cells on lymphoma cell proliferation].

Growing evidences show that mesenchymal stem cells (MSC) home to tumorgenesis and inhibit tumor cells, however, their molecular mechanisms are unclear. The purpose of this study was to explore the effect of B7-H4 in the influence of the mouse MSC on the proliferation of lymphoma cell line EL-4. The expression of B7-H4 on the MSC cell line C3H10T1/2 (C3H10) was detected by using immunofluorescence. FAM-siRNA was synthesized and transfected into C3H10 cells by INTERFER in (TM) siRNA Transfection Reagent. The transfection efficiency was determined by fluorescent microscopy and flow cytometry. The mRNA expression of B7-H4 was detected by RT-PCR after transfection of siRNA-B7-H4 into C3H10 cell line. The EL-4 was co-cultured with C3H10 siRNA-NC or C3H10 siRNA-B7-H4 for 48 hours, then was compared with EL-4 cultured alone, after 48 hours the cells were harvested under the confocal microscopy and measured by means of CCK-8 Kit. The results showed that the siRNA transfection efficiency in C3H10 cells reached to 72.43%, B7-H4 expressed highly on C3H10, the B7-H4 mRNA expression was down-regulated by transfection with different concentrations of siRNA into C3H10 cells. The proliferation of EL-4 was inhibited by C3H10 cells, and the effects were weakened and even disappeared after down-regulation of B7-H4. It is concluded that C3H10 can inhibit the proliferation of EL-4 through the expression of B7-H4, and this study provides new targets for the clinical treatment of lymphoma.

EZH2, a potential regulator of dental pulp inflammation and regeneration.

INTRODUCTION: Dental pulp has limited capability to regenerate, which happens in the early stage of pulpitis. An ambiguous relationship exists; inflammation may impair or support pulp regeneration. Epigenetics, which is involved in cell proliferation and inflammation, could regulate human dental pulp cell (HDPCs) regeneration. The aim of this study was to determine the role of the epigenetic mark, enhancer of zeste homolog 2 (EZH2), in the inflammation, proliferation, and regeneration of dental pulp. We used trimethylated histone H3 lysine 27(H3K27me3) and its lysine demethylase 6B (KDM6B) to monitor functional effects of altered EZH2 levels. METHODS: We detected epigenetic marks (EZH2, H3K27me3, and KDM6B) in pulp tissue by immunohistochemistry and immunofluorescence. EZH2 levels in HDPCs in inflammatory responses or differentiation were analyzed by quantitative polymerase chain reaction and Western blot. Quantitative polymerase chain reaction was used to assess the effects of EZH2 inhibition on interleukins in HDPCs upon tumor necrosis factor alpha stimulation. Cell proliferation was tested by cell counting kit-8, cell cycle, and apoptosis analysis. HDPC differentiation was investigated by quantitative polymerase chain reaction, alkaline phosphatase activity, and oil red O staining. RESULTS: EZH2 and H3K27me3 were decreased, whereas KDM6B was increased in infected pulp tissue and cells, which were similar to HDPC differentiation. EZH2 inhibition suppressed IL-1b, IL-6, and IL-8 messenger RNA (mRNA) in HDPCs upon inflammatory stimuli and impeded HDPC proliferation by decreasing cell number, arresting cell cycle, and increasing apoptosis. Suppressed EZH2 impaired adipogenesis, peroxisome proliferator-activated receptor r (PPAR-r), and CCAAT-enhancer binding protein a (CEBP/a) mRNA in adipogenic induction while enhancing alkaline phosphatase activity, Osx, and bone sialoprotein (BSP) mRNA in mineralization induction of HDPCs. CONCLUSIONS: EZH2 inhibited HDPC osteogenic differentiation while enhancing inflammatory response and proliferation, suggesting its role in pulp inflammation, proliferation, and regeneration.