EZH2 Mediates the Regulation of S100A4 on E-cadherin Expression and the Proliferation, Migration of Gastric Cancer Cells.

BACKGROUND/AIMS: Several reports have showed the inverse correlation between S100A4 and E-cadherin expression, but the exact molecular mechanism remained unclear. It has been reported that EZH2 mediates transcriptional silencing of E-cadherin by trimethylating lysine 27 of histone H3 (H3K27me3). Therefore, we hypothesized that EZH2 might mediate the inhibition of S100A4 on E-cadherin and further affect the functions of S100A4 in gastric cancer cells. METHODOLOGY: RT-PCR and Western Blot were used to detect the expression of EZH2 and E-cadherin after inhibiting or increasing S100A4 expression. MTT and Transwell assay were performed to detect the proliferation and migration of gastric cancer cells. RESULTS: Inhibition or overexpression of S100A4 led to decreased or increased EZH2 expression, and increased or decreased E-cadherin expression. The SET domain was important for EZH2 in rescuing the decreased proliferation and migration of the cells after S100A4 inhibition. CONCLUSION: As a novel downstream target of S100A4, EZH2 mediates the inhibition of S100A4 on E-cadherin. The SET domain is important for EZH2 in mediating the cellular function of S100A4.

MicroRNA-27a promotes proliferation and suppresses apoptosis by targeting PLK2 in laryngeal carcinoma.

BACKGROUND: miRNA-27a has been confirmed as an important regulator in carcinogenesis and other pathological processes. Whether and how it plays a role in the laryngeal carcinoma is unknown. METHODS: Mature miRNA-27a expression in laryngeal cancer was detected by qRT-PCR. Gain-of-function studies using mature miR-27a were performed to investigate cell proliferation and apoptosis in the Hep2 cells. In silico database analysis and luciferase reporter assay were applied to predict and validate the direct target, respectively. Loss-of-function assays were performed to investigate the functional significance of the miR-27a target gene. qRT-PCR and Western blot were used to evaluate mRNA and protein levels of the target, respectively. RESULTS: miR-27a was significantly up-regulated in the laryngeal tumor tissues compared to the adjacent non-tumor tissues. In silico database analysis result revealed that PLK2 is a potential target of miR-27a. luciferase reporter assay result showed the direct inhibition of miR-27a on PLK2-3'UTR. In the cases with miR-27a up-regulation, PLK2 protein expression level was significantly lower in cancer tissues than that in the adjacent non-tumor tissues, which showed a negative correlation with miR-27a expression level. Both miR-27a and knockdown of PLK2 caused the increase of the cell viability and colony formation and inhibition of the late apoptosis in the Hep2 cell lines. Moreover, miR-27a but not PLK2 also repressed the early apoptosis in the Hep2 cells. Additionally, no alteration of the Hep2 cell cycle induced by miR-27a was detected. CONCLUSIONS: miR-27a acts as an oncogene in laryngeal squamous cell carcinoma through down-regulation of PLK2 and may provide a novel clue into the potential mechanism of LSCC oncogenesis or serve as a useful biomarker in diagnosis and therapy in laryngeal cancer.

An interpretable clustering approach to safety climate analysis: Examining driver group distinctions.

The transportation industry, particularly the trucking sector, is prone to workplace accidents and fatalities. Accidents involving large trucks accounted for a considerable percentage of overall traffic fatalities. Recognizing the crucial role of safety climate in accident prevention, researchers have sought to understand its factors and measure its impact within organizations. While existing data-driven safety climate studies have made remarkable progress, clustering employees based on their safety climate perception is innovative and has not been extensively utilized in research. Identifying clusters of drivers based on their safety climate perception allows the organization to profile its workforce and devise more impactful interventions. The lack of utilizing the clustering approach could be due to difficulties interpreting or explaining the factors influencing employees' cluster membership. Moreover, existing safety-related studies did not compare multiple clustering algorithms, resulting in potential bias. To address these problems, this study introduces an interpretable clustering approach for safety climate analysis. This study compares five algorithms for clustering truck drivers based on their safety climate perceptions. It also proposes a novel method for quantitatively evaluating partial dependence plots (QPDP). Then, to better interpret the clustering results, this study introduces different interpretable machine learning measures (Shapley additive explanations, permutation feature importance, and QPDP). The Python code used in this study is available at https://github.com/NUS-DBE/truck-driver-safety-climate. This study explains the clusters based on the importance of different safety climate factors. Drawing on data collected from more than 7,000 American truck drivers, this study significantly contributes to the scientific literature. It highlights the critical role of supervisory care promotion in distinguishing various driver groups. Moreover, it showcases the advantages of employing machine learning techniques, such as cluster analysis, to enrich the scientific knowledge in this field. Future studies could involve experimental methods to assess strategies for enhancing supervisory care promotion, as well as integrating deep learning clustering techniques with safety climate evaluation.

Cross-Lingual Universal Dependency Parsing Only From One Monolingual Treebank.

Syntactic parsing is a highly linguistic processing task whose parser requires training on treebanks from the expensive human annotation. As it is unlikely to obtain a treebank for every human language, in this work, we propose an effective cross-lingual UD parsing framework for transferring parser from only one source monolingual treebank to any other target languages without treebank available. To reach satisfactory parsing accuracy among quite different languages, we introduce two language modeling tasks into the training process of dependency parsing as multi-tasking. Assuming only unlabeled data from target languages plus the source treebank can be exploited together, we adopt a self-training strategy for further performance improvement in terms of our multi-task framework. Our proposed cross-lingual parsers are implemented for English, Chinese, and 29 UD treebanks. The empirical study shows that our cross-lingual parsers yield promising results for all target languages, approaching the parser performance which is trained in its own target treebank.

Protocol for a distributed smart building solution using semi-physical simulation.

Honeycomb is a distributed smart building system that is robust, flexible, and portable. Here, we present a protocol that uses semi-physical simulation to construct a Honeycomb prototype. We describe steps for software and hardware preparation, as well as the implementation of a video-based occupancy detection algorithm. Besides, we provide examples and scenarios of distributed applications, including node failure and recovery. We further provide guidance on data visualization and analysis to facilitate the design of distributed applications for smart buildings. For complete details on the use and execution of this protocol, please refer to Xing et al.1.

Promoter hypermethylation-induced transcriptional down-regulation of the gene MYCT1 in laryngeal squamous cell carcinoma.

BACKGROUND: MYCT1, previously named MTLC, is a novel candidate tumor suppressor gene. MYCT1 was cloned from laryngeal squamous cell cancer (LSCC) and has been found to be down-regulated in LSCC; however, the regulatory details have not been fully elucidated. METHODS: Here, we sought to investigate the methylation status of the CpG islands of MYCT1 and mRNA levels by bisulfite-specific PCR (BSP) based on sequencing restriction enzyme digestion, reverse transcription and real-time quantitative polymerase chain reaction (RQ-PCR). The function of specific sites in the proximal promoter of MYCT1 in LSCC was measured by transient transfection, luciferase assays, electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation assay (ChIP). RESULTS: The results suggested hypermethylation of 12 CpG sites of the promoter in both laryngeal cancer tissues and the laryngeal cancer line Hep-2 cell. The hypermethylation of the site CGCG (-695 to -692), which has been identified as the c-Myc binding site, was identified in laryngeal cancer tissues (59/73) compared to paired mucosa (13/73); in addition, statistical analysis revealed that the methylation status of this site significantly correlated with cancer cell differentiation(p < 0.01). The mRNA level of MYCT1 increased in Hep-2 cells treated with 5-aza-C (p < 0.01). The luciferase activity from mutant transfectants pGL3-MYCT1m (-852/+12, mut-695-C > A, mut-693-C > G) was significantly reduced compared with the wild type pGL3-MYCT1 (-852/+12), while the luciferase activity from wild transfectants pGL3-MYCT1 (-852/+12) rose after 5-aza treatment in Hep-2 cells. Finally, EMSA and ChIP confirmed that the methylation of the CGCG (-695 to -692) site prevented c-Myc from binding of the site and demethylation treatment of the 5' flanking region of MYCT1 by 5-aza induced the increased occupation of the core promoter by c-Myc (p < 0.01). CONCLUSION: In summary, this study concluded that hypermethylation contributed to the transcriptional down-regulation of MYCT1 and could inhibit cancer cell differentiation in LSCC. DNA methylation of the CGCG site (-695 to -692) of MYCT1 altered the promoter activity by interfering with its binding to c-Myc in LSCC. Epigenetic therapy of reactivating MYCT1 by 5-aza should be further evaluated in clinical trails of LSCC.

Honeycomb: An open-source distributed system for smart buildings.

Restricted by the hierarchical and centralized system architecture, smart buildings face challenges such as limited adaptability and robustness, single application functionalities, and complex configurations. To address the above shortcomings, we learn from the activity patterns of natural bee swarms and propose Honeycomb, an open-source smart-building solution with fully distributed architecture. Honeycomb is a robust, flexible smart-building solution without any central server or global leader. An asynchronous leaderless spanning tree-based communication pattern is developed to generate and maintain the communication topology of Honeycomb in real time. Benefiting from this communication pattern, Honeycomb has plug-and-play ability. Various distributed applications are designed for building operating tasks and are deployed in a real Honeycomb prototype. The prototype demonstrates significant energy efficiency improvement from the control of the heating, ventilation, and air conditioning (HVAC) system with video-based occupancy information. Feedback on our Honeycomb prototype through questionnaires of users shows high acceptance of the controlled indoor environment.

MiR-125b targets BCL3 and suppresses ovarian cancer proliferation.

Micro-RNAs (miRNAs) important for post-transcriptional gene expression as negative regulators are endogenous 21- to 23-nucleotide noncoding RNAs. Many miRNAs are expressed in ovarian cancer (OC). In this study, we reported that miR-125b was underexpressed in human OC specimens. Ectopic expression of miR-125b in OC cells induced cell cycle arrest and led to reduction in proliferation and clonal formation. This inhibitory effect on OC cell growth was mediated by miR-125b inhibition of the translation of an mRNA encoding a proto-oncogene, BCL3. Furthermore, expression of miR-125b suppressed the tumor formation generated by injecting OC cells in nude mice. Our results suggest that aberrantly expressed miR-125b may contribute to OC development.

S100A4 protects gastric cancer cells from anoikis through regulation of αv and α5 integrin.

Detachment from the extracellular matrix induces a form of programmed cell death termed anoikis. Resistance to anoikis permits cancer cells to survive in systemic circulation and facilitates their metastasis to distant organs. It is well known that S100A4 is overexpressed in many tumors and involved in tumor metastasis, but the mechanisms of the metastasis-promoting function of S100A4 are not fully understood. We hypothesized that S100A4 might play a role in anoikis of gastric cancer cells and further affects their metastasis. To test this hypothesis, we changed the expression of S100A4 by means of RNA interference or experimental overexpression and investigated the effect on anoikis. We found that knockdown of S100A4 by RNA interference led to significantly increased anoikis, whereas overexpression of S100A4 resulted in inhibition of anoikis. Furthermore, we provide evidence that inhibition of S100A4 resulted in the downregulation of α5 and αv integrin expression. These findings suggest that S100A4 protects gastric cancer cells from anoikis by regulation of αv and α5 integrin expression, which sheds a novel light for the role of S100A4 in cancer metastasis.

Regulation of histone acetylation and nucleosome assembly by transcription factor JDP2.

Jun dimerization protein-2 (JDP2) is a component of the AP-1 transcription factor that represses transactivation mediated by the Jun family of proteins. Here, we examine the functional mechanisms of JDP2 and show that it can inhibit p300-mediated acetylation of core histones in vitro and in vivo. Inhibition of histone acetylation requires the N-terminal 35 residues and the DNA-binding region of JDP2. In addition, we demonstrate that JDP2 has histone-chaperone activity in vitro. These results suggest that the sequence-specific DNA-binding protein JDP2 may control transcription via direct regulation of the modification of histones and the assembly of chromatin.

Subcellular distribution of S100A4 and its transcriptional regulation under hypoxic conditions in gastric cancer cell line BGC823.

It is well known that S100A4 is overexpressed in many tumors and involved in tumor invasion and metastasis. But the regulation of it is ill understood. We previously found that hypoxia mimicking cobalt chloride (CoCl(2)) enhanced the mRNA and protein expressions of the S100A4 gene in the gastric cancer cell line BGC823. In this study we found that S100A4 also displayed increased expression in BGC823 cells after exposure to real hypoxia (2.5% O(2)) as that by CoCl(2) treatment. Moreover, S100A4 protein showed different subcellular distribution under real hypoxia compared with that by CoCl(2) treatment or in normoxic conditions. To investigate the underlying molecular mechanism by which hypoxia regulates the expression of S100A4, we analyzed the regulatory sequences of the genes by bioinformatics and found a putative hypoxia responsive element (HRE) motif in the first intron of S1004. Furthermore, luciferase reporter assay showed that it is responsive to hypoxia. Electrophoretic mobility shift assay and chromatin immunoprecipitation assays demonstrated that hypoxia-inducible factor 1 (HIF-1) binds to the functional HRE in vitro and in vivo. The results provide evidence that S100A4 is a hypoxia-inducible gene, whose transcription is stimulated at least partly through the interaction of HIF-1 and HRE located at +329 to +334 of S100A4.

14-3-3epsilon contributes to tumour suppression in laryngeal carcinoma by affecting apoptosis and invasion.

BACKGROUND: 14-3-3epsilon regulates a wide range of biological processes, including cell cycle control, proliferation, and apoptosis, and plays a significant role in neurogenesis and the formation of malignant tumours. However, the exact function and regulatory mechanism of 14-3-3epsilon in carcinogenesis have not been elucidated. METHODS: The expression of 14-3-3epsilon was assessed by RT-PCR and western blotting. The invasiveness and viability of Hep-2 cells were determined by the transwell migration assay and MTT assay, respectively. Cell cycle and apoptosis of Hep-2 cells were detected by flow cytometry. RESULTS: The mRNA and protein expression of 14-3-3epsilon in larynx squamous cell carcinoma (LSCC) tissues were significantly lower than those in clear surgical margin tissues. Statistical analysis showed that the 14-3-3epsilon protein level in metastatic lymph nodes was lower than that in paired tumour tissues. In addition, the protein level of 14-3-3epsilon in stage III or IV tumours was significantly lower than that in stage I or II tumours. Compared with control Hep-2 cells, the percentages of viable cells in the 14-3-3epsilon-GFP and negative control GFP groups were 36.68 +/- 14.09% and 71.68 +/- 12.10%, respectively. The proportions of S phase were 22.47 +/- 3.36%, 28.17 +/- 3.97% and 46.15 +/- 6.82%, and the apoptotic sub-G1 populations were 1.23 +/- 1.02%, 2.92 +/- 1.59% and 13.72 +/- 3.89% in the control, negative control GFP and 14-3-3epsilon-GFP groups, respectively. The percentages of the apoptotic cells were 0.84 +/- 0.25%, 1.08 +/- 0.24% and 2.93 +/- 0.13% in the control, negative control GFP and 14-3-3epsilon-GFP groups, respectively. The numbers of cells that penetrated the filter membrane in the control, negative control GFP and 14-3-3epsilon-GFP groups were 20.65 +/- 1.94, 17.63 +/- 1.04 and 9.1 +/- 0.24, respectively, indicating significant differences among the different groups. CONCLUSIONS: Decreased expression of 14-3-3epsilon in LSCC tissues contributes to the initiation and progression of LSCC. 14-3-3epsilon can promote apoptosis and inhibit the invasiveness of LSCC.

Reduced ACTC1 expression might play a role in the onset of congenital heart disease by inducing cardiomyocyte apoptosis.

BACKGROUND: The Cardiac α actin 1 gene (ACTC1) has been related to familial atrial septal defects. This study was set to explore a potential role of this gene in the formation of sporadic congenital heart disease (CHD). METHODS AND RESULTS: Assessment of cardiac tissue samples from 33 patients with sporadic CHD (gestational age (GA) 18 weeks-49 months) with real-time RT-PCR, Western blotting and immunohistochemistry has revealed a markedly decreased ACTC1 expression in the majority of samples (78.8%) compared with autopsied normal heart tissue from aged-matched subjects (GA 17 weeks-36 months). Also, as shown by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay, the proportion of apoptotic cardiomyocytes in samples featuring down-regulated ACTC1 expression (Group 1) was significantly greater than those with normal expression (Group 2) and the controls (P<0.01). The proportion of apoptotic cells strongly correlated with the expression of ACTC1 (r=-0.918, P<0.01). A study of 2 essential genes involved in apoptosis, Caspase-3 and Bcl-2, confirmed that the former has significantly increased expression, whilst the latter has decreased expression in Group 1 than in the other groups (P<0.01). Transfection of a small interfering RNA targeting, Actc1 (Actc1-siRNA), to a cardiomyocyte cell line, H9C2, also detected more apoptotic cells. CONCLUSIONS: Reduced ACTC1 expression might play a role in the onset of CHD through induction of cardiomyocyte apoptosis.

SH3GL2 gene participates in MEK-ERK signal pathway partly by regulating EGFR in the laryngeal carcinoma cell line Hep2.

BACKGROUND: The human Src homology 3 (SH3) domain GRB2-like 2 (SH3GL2) gene, a novel tumor suppressor gene in laryngeal squamous cell carcinoma (LSCC), induces apoptosis of tumor cells by regulating intra-cellular signal transduction networks. The objective of this study was to investigate the molecular mechanism of SH3GL2 in laryngeal carcinogenesis. MATERIAL/METHODS: RNA interference inhibited the expression of level of SH3GL, and RT-PCR and Western blotting were applied to evaluate the expression level of SH3GL2 after RNA interference. After RNA interference, flow cytometry and 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay were used to detect the biological effects, and Western blotting was used to determine the expression of EGFR and phosphorylated ERK1/2. The Hep2 cells transfected with siRNA-SH3GL2 were treated by U0126 (selective MEK1/2 Inhibitor), and the phosphorylated ERK1/2 proteins were detected by Western blotting; cell proliferation and apoptosis were detected subsequently. RESULTS: Our results show that the expression level of epidermal growth factor receptor (EGFR) and phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2) were up-regulated after down-regulation of SH3GL2. Additionally, SH3GL2 promoted apoptosis while decreasing cell proliferation. However, if ERK1/2 was inhibited by U0126, the apoptosis rate increased and proliferation decreased inversely. CONCLUSIONS: SH3GL2 participates in the regulation of apoptosis through the MEK-ERK signal pathway by adjusting EGFR in the laryngeal carcinoma cell line Hep2.

Low expression of Sp100 in laryngeal cancer: correlation with cell differentiation.

BACKGROUND: Sp100 is a permanent ProMyelocytic Leukaemian nuclear bodies (PML NB)-associated protein, and has been reported to participate in the regulation of transcriptional activity, apoptosis and other cellular biological processes. The aim of the present study was to explore the expression of Sp100 and its potential clinical implications in laryngeal cancer. MATERIAL/METHODS: The mRNA and protein levels of Sp100 in 96 laryngeal cancer samples and paired normal epithelium were examined by RT-PCR, Western blot and immuno-histochemical staining. The correlation of Sp100 expression with clinicopathological features of these patients was assessed by Chi-squared test. RESULTS: The coherent low Sp100 expression of both transcriptional and translational levels were confirmed in the malignant tissues compared to the normal mucosa, and the expression was down-regulated among the well-, moderately- and poorly-differentiated cancer cells accordingly. Moreover, immuno-histochemical staining demonstrated that Sp100 showed a predominantly nuclear pattern in well-differentiated cancer cells, and diffuse cellular distribution in cytoplasm in poorly-differentiated cancer cells. Besides histological type of cancer cells, other clinicopathological characteristics, including age, sex, T classification, lymph node metastasis, distant metastasis and clinical stage, showed no significant correlation with Sp100 low expression. CONCLUSIONS: Our finding provides an important clue to further understanding the role of Sp100 in the initiation and progression of tumorgenesis.

[Teaching experience in integrated course of human development and genetics].

Establishment of integrated course system in human development and genetics is an important part of course reformation, and the improvement of this system is achieved by integrating the content of course, stabilizing teaching force, building teaching materials and applying problem-based learning. Integrity-PBL teaching model is founded and proved to be feasible and effective by teaching practice. Therefore, it maybe play an important role in improving teaching effect and cultivating ability of students to analyse and solve problems.

[The mechanism of TBX5 abnormal expression in simple congenital heart disease].

To explore the mechanism of TBX5 abnormal expression in simple congenital heart disease (CHD), 100 CHD venous blood, 50 CHD heart tissues, and 5 non-CHD heart tissues were involved in this study. The mutation and methylation in the 1 200 bp region upstream of TBX5 gene were detected by high-performance liquid chromatography (DHPLC) and methylation-sensitive restriction endonuclease (MS-RE), respectively. The binding site of NKX2-5 to Tbx5 predicted by P-MATCH software was validated by EMSA (Electrophoretic mobility shift assay). Tbx5 gene expression in mouse cardiac muscle cell H9C2(2-1) transfected with NKX2-5 expression vector was evaluated. No mutation was found in all patients. Both non-CHD and CHD heart tissues had the same methylation in the two CpG islands. Exogenous Nkx2-5 efficiently activated the transcription of the endogenous Tbx5 gene in H9C2 (2-1) cells. EMSA showed that the special binding band appeared when Nkx2-5 existed. These results indicates that the down expression of TBX5 might not be caused by mutation and methylation in the 1 200 bp region upstream of gene, and might be regulated by abnormal expression of NKX2-5 gene in heart muscle of CHD.

HLA-B gene participates in the NF-kappaB signal pathway partly by regulating S100A8 in the laryngeal carcinoma cell line Hep2.

Human leukocyte antigen B (HLA-B), a novel member of the NF-kappaB signal pathway in laryngeal squamous cell carcinoma (LSCC), mediates immunological surveillance of tumor cells by presenting peptides to cytotoxic T-lymphocytes (CTLs) together with S100 calcium binding protein A8 (S100A8). The objective of this study was to investigate the molecular mechanism of HLA-B and S100A8 in laryngeal carcinogenesis. Flow cytometry, 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay and cell invasion in vitro were used to detect the biological effect of the Hep2 cell line induced by HLA-B RNA interference. RT-PCR and Western blotting were applied to evaluate the expression level of the S100A8 gene after HLA-B RNA interference. Our results showed that HLA-B had negative effects on Hep2 cells by inhibiting apoptosis and cell invasion while decreasing cell proliferation. Additionally, the expression level of HLA-B and S100A8 in LSCC were down-regulated after HLA-B RNA interference. The abnormal expression of HLA-B is thus relevant to the biological effect of laryngeal carcinoma and participates in the NF-kappaB signal pathway partly by regulating the expression of the S100A8 gene.

[Research of HOXD13 and FHL1 in idiopathic congenital talipes equinovarus].

To investigate the relationship of HOXD13 and FHL1 in idiopathic congenital talipes equinovarus(ICTEV), 84 samples from patients with ICTEV were used in the study. Mutation in the coding region of HOXD13 was detected by denaturing gradinent electrophoresis. The mRNA and protein levels of HOXD13 and FHL1 were evaluated by RT-PCR and immunohistochemistry, respectively. The binding site of FHL1 to HOXD13 predicted by PMATCH software was validated by EMSA( Electrophoretic mobility shift assay,EMSA).No mutation was found in the coding region of HOXD13 in 84 samples from patients with ICTEV. Both HOXD13(33.3%) and FHL1(46.6%) were down-regulated in ICTEV muscle tissue. The result of EMSA showed that the special binding band appeared when HOXD13 existed. The results shows that HOXD13 gene mutation was not involved in outbreak in idiopathic congenital talipes equinovarus, but changes of HOXD13 and FHL1 gene expression related to the development of talipes equinovarus malformation. HOXD13 might play an role in ICTEV through regulating FHL1 expression.

[Association study between mutations of transcription regulator sequences of COL1A1 gene and idiopathic con-genital talipes equinovarus].

RT-PCR was used to detect the expressions of COL1A1 mRNA in 20 patients with idiopathic congenital talipes equinovarus (ICTEV). The primers were designed by Primer 5 according to sequences of -1 031 bp~ +30 bp and the first intron of COL1A1. PCR-DGGE was used to screen the mutations in COL1A1 gene. Expression of COL1A1 on mRNA levels showed significantly higher in patients with ICTEV than in normal persons (t=12.680, P < 0.05). By DNA sequencing, a -161(T--> C) heterozygous mutation and a+ 274(C-->G) homozygous mutation were detected, and both were new identified mutations. These results indicated that the mutations in transcription regulator sequences of COL1A1 could cause ICTEV.