S670, an amide derivative of 3-O-acetyl-11-keto-ß-boswellic acid, induces ferroptosis in human glioblastoma cells by generating ROS and inhibiting STX17-mediated fusion of autophagosome and lysosome.

Glioblastoma (GBM) is the most common malignant tumor in the brain with temozolomide (TMZ) as the only approved chemotherapy agent. GBM is characterized by susceptibility to radiation and chemotherapy resistance and recurrence as well as low immunological response. There is an urgent need for new therapy to improve the outcome of GBM patients. We previously reported that 3-O-acetyl-11-keto-ß-boswellic acid (AKBA) inhibited the growth of GBM. In this study we characterized the anti-GBM effect of S670, a synthesized amide derivative of AKBA, and investigated the underlying mechanisms. We showed that S670 dose-dependently inhibited the proliferation of human GBM cell lines U87 and U251 with IC50 values of around 6 µM. Furthermore, we found that S670 (6 µM) markedly stimulated mitochondrial ROS generation and induced ferroptosis in the GBM cells. Moreover, S670 treatment induced ROS-mediated Nrf2 activation and TFEB nuclear translocation, promoting protective autophagosome and lysosome biogenesis in the GBM cells. On the other hand, S670 treatment significantly inhibited the expression of SXT17, thus impairing autophagosome-lysosome fusion and blocking autophagy flux, which exacerbated ROS accumulation and enhanced ferroptosis in the GBM cells. Administration of S670 (50 mg·kg-1·d-1, i.g.) for 12 days in a U87 mouse xenograft model significantly inhibited tumor growth with reduced Ki67 expression and increased LC3 and LAMP2 expression in the tumor tissues. Taken together, S670 induces ferroptosis by generating ROS and inhibiting STX17-mediated fusion of autophagosome and lysosome in GBM cells. S670 could serve as a drug candidate for the treatment of GBM.

Progress of studies on natural products for glioblastoma therapy.

Glioblastoma (GBM) is the most common, malignant, and lethal primary brain tumor in adults. Up to now, the chemotherapy approaches for GBM are limited. Therefore, more studies on identifying and exploring new chemotherapy drugs or strategies overcome the GBM are essential. Natural products are an important source of drugs against various human diseases including cancers. With the better understanding of the molecular etiology of GBM, the development of new anti-GBM drugs has been increasing. Here, we summarized recent researches of natural products for the GBM therapy and their potential mechanisms in details, which will provide new ideas for the research on natural products and promote developing drugs from nature products for GBM therapy.

Tspan5 promotes the EMT process to regulate the syncytialization of trophoblast cells by activating Notch signalling.

Placental trophoblastic cells play important roles in placental development and fetal health. However, the mechanism of trophoblastic cell fusion is still not entirely clear. The level of Tspan5 in the embryo culture medium was detected using enzyme-linked immunosorbent assay (ELISA). Fusion of BeWo cells was observed by immunofluorescence. Cell fusion-related factors and EMT-related factors were identified by qRT-PCR and western blotting. Notch protein repressor DAPT was used to verify the role of Tspan5 in BeWo cells. The expression of Tspan5 was significantly increased in embryo culture medium. The fusion of BeWo cells was observed after treatment with forskolin (FSK). Cell fusion-related factors (i.e. ß-hCG and syncytin 1/2) and Tspan5 were significantly increased after FSK treatment. In addition, FSK treatment promoted EMT-related protein expression in BeWo cells. Knockdown of Tspan5 inhibited cell fusion and EMT-related protein levels. Notch-1 and Jagged-1 protein levels were significantly upregulated, and the EMT process was activated by overexpression of Tspan5 in FSK-treated BeWo cells. Interestingly, blocking the Notch pathway by the repressor DAPT had the opposite results. These results indicated that Tspan5 could promote the EMT process by activating the Notch pathway, thereby causing cell fusion. These findings contribute to a better understanding of trophoblast cell syncytialization and embryonic development. Tspan5 may be used as a therapeutic target for normal placental development.

[CT-Based Weighted Radiomic Score Predicts Tumor Response to Immunotherapy in Non-Small Cell Lung Cancer].

Objective To develop a CT-based weighted radiomic model that predicts tumor response to programmed death-1(PD-1)/PD-ligand 1(PD-L1)immunotherapy in patients with non-small cell lung cancer.Methods The patients with non-small cell lung cancer treated by PD-1/PD-L1 immune checkpoint inhibitors in the Peking Union Medical College Hospital from June 2015 to February 2022 were retrospectively studied and classified as responders(partial or complete response)and non-responders(stable or progressive disease).Original radiomic features were extracted from multiple intrapulmonary lesions in the contrast-enhanced CT scans of the arterial phase,and then weighted and summed by an attention-based multiple instances learning algorithm.Logistic regression was employed to build a weighted radiomic scoring model and the radiomic score was then calculated.The area under the receiver operating characteristic curve(AUC)was used to compare the weighted radiomic scoring model,PD-L1 model,clinical model,weighted radiomic scoring + PD-L1 model,and comprehensive prediction model.Results A total of 237 patients were included in the study and randomized into a training set(n=165)and a test set(n=72),with the mean ages of(64±9)and(62±8)years,respectively.The AUC of the weighted radiomic scoring model reached 0.85 and 0.80 in the training set and test set,respectively,which was higher than that of the PD-L1-1 model(Z=37.30,P<0.001 and Z=5.69,P=0.017),PD-L1-50 model(Z=38.36,P<0.001 and Z=17.99,P<0.001),and clinical model(Z=11.40,P<0.001 and Z=5.76,P=0.016).The AUC of the weighted scoring model was not different from that of the weighted radiomic scoring + PD-L1 model and the comprehensive prediction model(both P>0.05).Conclusion The weighted radiomic scores based on pre-treatment enhanced CT images can predict tumor responses to immunotherapy in patients with non-small cell lung cancer.

ADH1C inhibits progression of colorectal cancer through the ADH1C/PHGDH /PSAT1/serine metabolic pathway.

Colorectal cancer (CRC) is the third most common cancer in men and the second most common cancer in women worldwide. CRC is the second leading cause of cancer-related deaths. Although some progress in the treatment of CRC has been achieved, the molecular mechanism of CRC is still unclear. In this study, alcohol dehydrogenase 1C(ADH1C) was first identified as a target gene closely associated with the development of CRC by the comprehensive application of transcriptomics, proteomics, metabonomics and in silico analysis. The ADH1C mRNA and protein expression in CRC cell lines and tumor tissues was lower than that in normal intestinal epithelial cell lines and healthy tissues. Overexpression of ADH1C inhibited the growth, migration, invasion and colony formation of CRC cell lines and prevented the growth of xenograft tumors in nude mice. The inhibitory effects of ADH1C on CRC cells in vitro were exerted by reducing the expression of PHGDH/PSAT1 and the serine level. This inhibition could be partially reversed by adding serine to the culture medium. These results showed that ADH1C is a potential drug target in CRC.

Benzimidazoles induce concurrent apoptosis and pyroptosis of human glioblastoma cells via arresting cell cycle.

Glioblastoma multiforme (GBM) is the most malignant and lethal primary brain tumor in adults accounting for about 50% of all gliomas. The only treatment available for GBM is the drug temozolomide, which unfortunately has frequent drug resistance issue. By analyzing the hub genes of GBM via weighted gene co-expression network analysis (WGCNA) of the cancer genome atlas (TCGA) dataset, and using the connectivity map (CMAP) platform for drug repurposing, we found that multiple azole compounds had potential anti-GBM activity. When their anti-GBM activity was examined, however, only three benzimidazole compounds, i.e. flubendazole, mebendazole and fenbendazole, potently and dose-dependently inhibited proliferation of U87 and U251 cells with IC50 values below 0.26 µM. Benzimidazoles (0.125-0.5 µM) dose-dependently suppressed DNA synthesis, cell migration and invasion, and regulated the expression of key epithelial-mesenchymal transition (EMT) markers in U87 and U251 cells. Benzimidazoles treatment also dose-dependently induced the GBM cell cycle arrest at the G2/M phase via the P53/P21/cyclin B1 pathway. Furthermore, the drugs triggered pyroptosis of GBM cells through the NF-κB/NLRP3/GSDMD pathway, and might also concurrently induced mitochondria-dependent apoptosis. In a nude mouse U87 cell xenograft model, administration of flubendazole (12.5, 25, and 50 mg · kg-1 · d-1, i.p, for 3 weeks) dose-dependently suppressed the tumor growth without obvious adverse effects. Taken together, our results demonstrated that benzimidazoles might be promising candidates for the treatment of GBM.

Apolipoprotein C1 promotes glioblastoma tumorigenesis by reducing KEAP1/NRF2 and CBS-regulated ferroptosis.

Glioblastoma (GBM), a malignant brain tumor, is a world-wide health problem because of its poor prognosis and high rates of recurrence and mortality. Apolipoprotein C1 (APOC1) is the smallest of apolipoproteins, implicated in many diseases. Recent studies have shown that APOC1 promotes tumorigenesis and development of several types of cancer. In this study we investigated the role of APOC1 in GBM tumorigenesis. Using in silico assays we showed that APOC1 was highly expressed in GBM tissues and its expression was closely related to GBM progression. We showed that APOC1 protein expression was markedly increased in four GBM cell lines (U251, U138, A172 and U87) compared to the normal brain glia cell lines (HEB, HA1800). In U251 cells, overexpression of APOC1 promoted cell proliferation, migration, invasion and colony information, which was reversed by APOC1 knockdown. APOC1 knockdown also markedly inhibited the growth of GBM xenografts in the ventricle of nude mice. We further demonstrated that APOC1 reduced ferroptosis by inhibiting KEAP1, promoting nuclear translocation of NRF2 and increasing expression of HO-1 and NQO1 in GBM cells. APOC1 also induced ferroptosis resistance by increasing cystathionine beta-synthase (CBS) expression, which promoted trans-sulfuration and increased GSH synthesis, ultimately leading to an increase in glutathione peroxidase-4 (GPX4). Thus, APOC1 plays a key role in GBM tumorigenesis, conferring resistance to ferroptosis, and may be a promising therapeutic target for GBM.

Insertion/deletion polymorphism at angiotensin-converting enzyme gene in PTSD individuals and their reciprocal effects on blood pressure.

OBJECTIVES: The aim of the present study was to investigate relationships between insertion/deletion (I/D) polymorphism at angiotensin-converting enzyme gene (ACE) and post-traumatic stress disorder (PTSD), as well as their interactions on blood pressure. METHODS: Variants of ACE I/D were identified by polymerase chain reaction method and verified by DNA sequencing. PTSD symptoms were assessed by the PTSD Checklist-Civilian Version (PCL-C) based on DSM-IV-TR criteria among high school students at 6 months after the 2008 Wenchuan earthquake. RESULTS: Female subjects were found to have higher prevalence of PTSD and PCL-C scores than male counterparts in the II homozygotes (p = .038 for PTSD and p = .003 for PCL-C scores) and the ID heterozygotes (p = .000 for PTSD and p = .000 for PCL-C scores), but not in the DD homozygotes. Male subjects with the ID (p = .046) or the DD genotype (p = .039) had lower pulse pressure (PP) than the male II homozygotes, while the female II homozygotes had lower diastolic blood pressure (DBP) than the female DD homozygotes (p = .036). ACE I/D, PTSD, or PCL-C scores, as well as gender and BMI, were found to be the predictors of PP. CONCLUSIONS: These results indicate that there are interactions of ACE I/D and PTSD, together with gender and BMI, on PP. This finding may be the additional explanation for the heterogeneous relationships between PTSD and blood pressure, and suggest psychiatry care and different medication strategies for patients with comorbidities of PTSD and hypertension and with different genotypes of ACE I/D.

6-Shogaol from ginger shows anti-tumor effect in cervical carcinoma via PI3K/Akt/mTOR pathway.

PURPOSE: 6-Shogaol, an active phenolic compound from ginger (Zingiber officinale), can inhibit the growth of a variety of human cancer cells. Nevertheless, its underlying molecular mechanisms in cervical cancer remain unclear. In this study, we systematically examine the inhibitory effect of 6-shogaol on cervical cancer in vitro and in vivo. METHODS: Cell proliferation was assessed by CCK8 assay and colony formation assay in HeLa and SiHa cells. We analyzed cell cycle and apoptosis through flow cytometry. GFP-LC3 puncta and transmission electron microscopy were used to observe autophagic bodies. Wound-healing assay and transwell assay were used for evaluating the migration of cells. Western blot was applied to detect protein expression levels. RESULTS: 6-Shogaol could suppress cell proliferation and migration, cause cell cycle arrest in the G2/M phase in HeLa and SiHa cells. Moreover, 6-shogaol triggered the apoptosis process through the mitochondrial pathway by downregulating the expression levels of p-PI3K, p-Akt and p-mTOR. Further research indicated that the induction of apoptosis by 6-shogaol was remarkably decreased after the treatment of ROS scavenger and PI3K agonist. Additionally, 6-shogaol increased the number of LC3-positive puncta and autophagic bodies per cell in both HeLa and SiHa cells. Pretreatment of cells with Bafilomycin A1, an autophagy inhibitor, accelerated 6-shogaol mediated cell apoptosis, suggesting that induction of autophagy by 6-shogaol is suppressive to apoptosis. Furthermore, in vivo data revealed that 6-shogaol significantly inhibited tumor growth and cell proliferation in tumor tissues. CONCLUSION: These findings suggested that 6-shogaol could be developed as a functional food ingredient, which is potentially used as therapeutic agents for patients with cervical cancer.

Remarkable inhibition effects of afatinib alone or combining with paclitaxel in esophageal squamous cell carcinoma.

BACKGROUND AND AIM: Chemotherapy drugs do not work well in esophageal squamous cell carcinoma (ESCC), and none of the targeted drugs have been applied in clinic. This study aims to identify effective targeted drugs and related biomarkers for the treatment of ESCC. METHODS: The effect of 40 Food and Drug Administration-approved small-molecule inhibitors was first tested in five ESCC cell lines. CCK8 assays and xenografts derived from ESCC cell lines were performed to evaluate the anti-ESCC effects of inhibitors or chemotherapeutic agents in vitro and in vivo, respectively. Immunohistochemistry was utilized to analyze the p-EGFR expression in tissues. Western blot combining with gray analysis was conducted to detect the expression of interest protein. Flow cytometry and immunofluorescence assay were used to analyze apoptosis, cell cycle, and mitotic changes after drug treatment. RESULTS: Afatinib showed remarkable effects on inhibiting ESCC cells with higher expression of p-EGFR. Results from combinatorial screening in ESCC cells expressing lower phosphorylation level of EGFR showed that paclitaxel and afatinib presented a significant synergistic inhibitory effect (P < 0.001). Molecular analysis revealed that paclitaxel sensitized afatinib by activating EGFR, and afatinib in combination with paclitaxel effectively blocked MAPK pathway and induced G2/M cell arrest and apoptosis that is an indicator of mitotic catastrophe. CONCLUSIONS: Our data demonstrate that afatinib is an effective drug for patients with ESCC expressing higher phosphorylation level of EGFR. And for patients with lower p-EGFR in tumors, paclitaxel in combination with afatinib might be a promising therapeutic strategy in ESCC.

Avasimibe exerts anticancer effects on human glioblastoma cells via inducing cell apoptosis and cell cycle arrest.

Glioblastoma (GBM) is the most common and lethal primary brain tumor in adults, but there is no effective drug available for GBM. Avasimibe is a potent inhibitor of acyl-coenzyme A: cholesterol acyltransferase-1 (ACAT-1), which was used to treat atherosclerosis. Experimental evidence and bioinformatics have shown that avasimibe has anticancer activity. In this study we investigated the anticancer effects of avasimibe on human glioblastoma cells and the underlying mechanisms. Our results showed that avasimibe dose-dependently inhibited the proliferation of U251 and U87 human glioblastoma cells with IC50 values of 20.29 and 28.27 µM, respectively, at 48 h. Avasimibe (7.5, 15, 30 µM) decreased the DNA synthesis, and inhibited the colony formation of the tumor cells. Treatment of avasimibe also dose-dependently increased the apoptotic rate of tumor cells, decreased the mitochondrial membrane potential, induced the activity of caspase-3/7, and increased the protein expression of cleaved caspase-9, cleaved PARP and Bax in U251 and U87 cells. RNA-sequencing analyses revealed that avasimibe suppressed the expression of CDK2, cyclin E1, CDK4, cyclin D, CDK1, cyclin B1, Aurora A, and PLK1, while induced the expression of p53, p21, p27, and GADD45A, which was validated by Western blot analysis. These results demonstrated that avasimibe induced mitochondria-dependent apoptosis in glioblastoma cells, which was associated with arresting the cell cycle at G0/G1 phase and G2/M phase by regulating the p53/p21 pathway, p53/GADD45A and Aurora A/PLK1 signaling pathways. In U87 xenograft nude mice model, administration of avasimibe (15, 30 mg·kg-1·d-1, ip, for 18 days) dose-dependently inhibit the tumor growth. Taken together, our results demonstrated that avasimibe might be a promising chemotherapy drug in the treatment of GBM.

EZH2 regulates expression of FOXC1 by mediating H3K27me3 in breast cancers.

Triple-negative breast cancer (TNBC) is characterized by low expression of human epidermal growth factor receptor-2 (HER2), estrogen receptor (ER), and progesterone receptor (PR), which is the most aggressive subtype with poor outcome among breast cancers. The underlying mechanisms of TNBC remain unclear and there is a lack of biomarkers. In this study we conducted an in silico assay and found that FOXC1 was highly expressed in ER-/PR-/HER2- breast cancers, which was confirmed by qRT-PCR, immunohistochemistry, and Western blot analysis. FOXC1 was more highly expressed in TNBCs than the other breast cancers. Kaplan-Meier plotter revealed that expression of FOXC1 was associated with overall survival (OS) of patients with breast cancers. Expression of FOXC1 was reversely associated with level of H3K27me3, which was methylated by EZH2. In MCF-7 and T47D cells, inhibition of EZH2 by DZNeP or GSK343 concentration- and time-dependently increased expression of FOXC1. Finally, we demonstrated that the expression of FOXC1 was associated with resistance of doxorubicin treatment of breast cancer cells. In conclusion, these results suggest that FOXC1 may be a potential biomarker or drug target for TNBCs, and that downregulation of FOXC1 could have therapeutic value in treatment of TNBCs.

Dynamic monocyte chemoattractant protein-1 level as predictors of perceived pain during first and second phacoemulsification eye surgeries in patients with bilateral cataract.

BACKGROUND: The purpose of the study was to investigate whether dynamic monocyte chemoattractant protein-1 (MCP-1) level might be as predictors of perceived pain during the first and second phacoemulsification eye surgeries in patients with bilateral cataract. METHODS: Consecutive bilateral cataract patients undergoing bilateral sequential phacoemulsification were retrospectively enrolled. Patients' preoperative anxiety score and intraoperative pain score were registered. Aqueous humor samples were obtained during surgery. MCP-1 level in the aqueous humor was measured by enzyme linked immunosorbent assay (Elisa). Patients were assigned to seven subgroups based on the interval between first-eye and second-eye cataract surgery. Comparisons were performed for a subjective sensation and MCP-1 levels among different subgroups. RESULTS: pain score during second-eye surgery was significantly higher than during first-eye surgery. Whereas there was no statistical difference in anxiety score between both surgeries. Result from subgroups comparison showed that the visual analog scale (VAS) pain score was statistically greater in 1-group and 6-group during the second eye surgery. Anxiety score did not statistically differ in subgroups. Additionally, the second-eye MCP-1 level was significantly higher at week 1and 6 intervals. Preoperative MCP-1 level was positively correlated with perceiving pain score during both surgeries. CONCLUSIONS: MCP-1 level in aqueous humor significantly correlated with perceived pain during cataract surgery. Dynamic MCP-1 level could function as predictors of perceived pain during the first and second phacoemulsification eye surgeries in patients with bilateral cataract, which might support clinicians in treatment optimization and clinical decision-making.

Validation of the Chinese version of the Malocclusion Impact Questionnaire (MIQ).

OBJECTIVE: To translate and cross-culturally adapt the Malocclusion Impact Questionnaire (MIQ) into Chinese and to assess the psychometric properties of the Chinese version of the MIQ (MIQ/C) for use among adolescents with malocclusion in China. MATERIALS AND METHOD: First, the MIQ/C was developed according to international guidelines. Then, the MIQ/C was filled out by 536 adolescents between 10 and 16 years of age. This study used exploratory factor analysis (EFA), confirmatory factor analysis (CFA), convergent validity, internal consistency, and test-retest reliability to evaluate the psychometric properties of the MIQ/C. RESULTS: Following EFA, three domains were extracted, accounting for 65.950% of the total variance. The CFA results showed that the fit indices of each factor in the three-factor model all reached the standard (chi-square/DF = 2.591, GFI = 0.919, TLI = 0.926, CFI = 0.928, RMSEA = 0.076). The scale evidenced a good relationship with the two global questions, indicating good convergent validity. The Cronbach alpha value and the intraclass correlation coefficients (ICC) value of the MIQ/C were 0.929 and 0.893, respectively. CONCLUSION: The MIQ/C demonstrated good reliability and validity and can be further studied and applied in Chinese adolescents with malocclusion. CLINICAL RELEVANCE: The MIQ/C can be applied to assess the psychosocial impact of malocclusion among Chinese adolescents.

Determining the minimal important difference of the Oral Health Impact Profile for Chronic Periodontitis.

AIM: Building on previous psychometric work, we aimed to further assess the minimally important difference (MID) of the Oral Health Impact Profile for Chronic Periodontitis (OHIP-CP). METHODS: In total, 240 consecutive patients with chronic periodontitis were recruited in the study. The OHIP-CP was completed at baseline and after six weeks. Methodology testing included the confirmatory factor analysis (CFA) and MID. Confirmatory factor analysis (CFA) was performed to assess the fit of the previously proposed three-factor model. The MID of this questionnaire was determined by applying anchor-based and distribution-based approaches. RESULTS: The CFA supported a three-factor model for the OHIP-CP with acceptable fit to the data. The fit indices were χ2 /df = 2.231, GFI = 0.935, TLI = 0.969 and CFI = 0.976, RMSEA = 0.076. The OHIP-CP scores showed significant improvements after treatment (p < .001). The anchor-based MIDs of OHIP-CP for "oral function restriction," "oral pain" and "psychological and social impact," and total score were 2, 1, 4 and 7 points, respectively. The effect sizes (ES) and standardized response mean (SRM) for the OHIP-CP were moderate to large. CONCLUSIONS: The MID of the OHIP-CP is recommended for interpreting clinically meaningful change in oral health-related quality of life (OHRQoL) over time.

OHRQoL changes among Chinese preschool children following dental treatment under general anesthesia.

OBJECTIVE: To assess dental treatment under dental general anesthesia (DGA) among Chinese preschool children by investigating changes in their oral health-related quality of life (OHRQoL), the incidence of postoperative complications, and parental satisfaction. METHOD: A single-center prospective cohort study was conducted. A total of 190 children who received treatment for early childhood caries (ECC) under DGA were included. The primary outcome was a change in the children's OHRQoL at 1 month after the operation compared to that at baseline, which was measured by the Early Childhood Oral Health Impact Scale (ECOHIS). The secondary outcomes were the incidence of complications within 1 day after treatment and parental satisfaction with the DGA treatment. RESULTS: In total, 180 participants were successfully reevaluated after the operation, yielding a 94.7% follow-up response rate. The total ECOHIS score decreased by 76.3% (P < 0.01) after treatment, demonstrating a large effect. Approximately 74.4% of the children complained of at least one complication, including sleepiness (43.3%), emergence agitation (38.9%), nausea/vomiting (13.9%), dizziness (10.6%), and fever (3.3%), on the first day. Approximately 85.5% of the parents were satisfied with the DGA treatment. CONCLUSION: DGA treatment has a positive effect on Chinese preschool children's OHRQoL and is evaluated highly by their parents. CLINICAL RELEVANCE: Dental treatment under general anesthesia improved the OHRQoL of Chinese preschool children.

Validation of a web-based version of the Craniofacial Pain and Disability Inventory.

OBJECTIVE: To translate and cross-culturally adapt the Craniofacial Pain and Disability Inventory (CF-PDI) into Chinese and, to evaluate measurement properties of the web-based version of the CF-PDI. METHODS: The Chinese version (CF-PDI/C) was first produced according the guidelines. Then, the web-based version of CF-PDI/C (eCF-PDI/C) was developed by our team and a third-party survey provider. The eCF-PDI/C was distributed to patients with painful temporomandibular disorders (TMD) with or without headache to evaluate its psychometric properties. The reliability of the eCF-PDI/C was detected by internal consistency and test-retest reliability. The validity of the measure was performed through construct validity and convergent validity. RESULTS: A total of 206 patients were recruited. The Cronbach's α value and the intraclass correlation coefficients (ICC) value of the eCF-PDI/C was .912 and .82, respectively. Following exploratory factor analysis (EFA), three factors were extracted, accounting for 77.153% of the total variation. With regard to the convergent validity, the measure evidenced a good relationship with the global health question. CONCLUSIONS: The eCF-PDI/C displays good reliability and validity through rigorous performance tests. This can be recommended for use among patients with painful TMD with or without headache.

CRISPR/Cas9 facilitates genomic editing for large-scale functional studies in pluripotent stem cell cultures.

Pluripotent stem cell (PSC) cultures form an integral part of biomedical and medical research due to their capacity to rapidly proliferate and differentiate into hundreds of highly specialized cell types. This makes them a highly useful tool in exploring human physiology and disease. Genomic editing of PSC cultures is an essential method of attaining answers to basic physiological functions, developing in vitro models of human disease, and exploring potential therapeutic strategies and the identification of drug targets. Achieving reliable and efficient genomic editing is an important aspect of using large-scale PSC cultures. The CRISPR/Cas9 genomic editing tool has facilitated highly efficient gene knockout, gene correction, or gene modifications through the design and use of single-guide RNAs which are delivered to the target DNA via Cas9. CRISPR/Cas9 modification of PSCs has furthered the understanding of basic physiology and has been utilized to develop in vitro disease models, to test therapeutic strategies, and to facilitate regenerative or tissue repair approaches. In this review, we discuss the benefits of the CRISPR/Cas9 system in large-scale PSC cultures.