Fever burden within 24 h after hematoma evacuation predicts early neurological deterioration in patients with intracerebral hemorrhage: a retrospective analysis.

Background: Early neurological deterioration after hematoma evacuation is closely associated with a poor prognosis in patients with intracerebral hemorrhage. However, the relationship between body temperature after hematoma evacuation and early neurological deterioration remains unclear. Therefore, this study aims to explore the possible relationship between body temperature and early neurological deterioration in patients with intracerebral hemorrhage after hematoma evacuation. Methods: We retrospectively collected data from patients with cerebral hemorrhage at our institute between January 2017 and April 2022. The Student's t-test, Mann-Whitney U-test, and χ2 Test and Fisher's exact test were used to analyze the clinical baseline data. A univariate logistic regression model was used to evaluate the association between the body temperature indices and early neurological deterioration. The predictive power was assessed using the area under the Receiver Operating Characteristic (ROC) curve. The secondary outcome was a poor functional outcome. Results: Among 2,726 patients with intracerebral hemorrhage, 308 who underwent hematoma evacuation were included in the present analysis. A total of 82 patients (22.6%) developed early neurological deterioration. Univariate analysis showed that sex (p = 0.041); body temperature at 6 h (p = 0.005), 12 h (p = 0.01), and 24 h (p = 0.008) after surgery; duration of fever (p = 0.008); and fever burden (p < 0.001) were associated with early neurological deterioration. Multivariate logistic regression showed that fever burden was independently associated with early neurological deterioration (OR = 1.055 per °C × hour, 95%CI 1.008-1.103, p = 0.020). ROC showed that fever burden (AUC = 0.590; 95%CI: 0.514-0.666) could predict the occurrence of early neurological deterioration. Conclusion: Fever burden is associated with early neurological deterioration in intracerebral hemorrhage patients undergoing hematoma evacuation. Our findings add to previous evidence on the relationship between the fever burden and the occurrence of early neurological deterioration in patients with intracerebral hemorrhage. Future studies with larger sample sizes are required to confirm these findings.

Development and Validation of a Nomogram to Predict Frailty Progression in Nonfrail Chinese Community-Living Older Adults.

OBJECTIVE: Frailty state progression is common among older adults, so it is necessary to identify predictors to implement individualized interventions. We aimed to develop and validate a nomogram to predict frailty progression in community-living older adults. DESIGN: Prospective cohort study. SETTING AND PARTICIPANTS: A total of 3170 Chinese community-living people aged ≥60 years were randomly assigned to a training set or validation set at a ratio of 6:4. METHODS: Candidate predictors (demographic, lifestyle, and medical characteristics) were used to predict frailty state progression as measured with the Fried frailty phenotype at a 4-year follow-up, and multivariate logistic regression analysis was conducted to develop a nomogram, which was validated internally with 1000 bootstrap resamples and externally with the use of a validation set. The C index and calibration plot were used to assess discrimination and calibration of the nomogram, respectively. RESULTS: After a follow-up period of 4 years, 64.1% (917/1430) of the participants in the robust group and 26.0% (453/1740) in the prefrail group experienced frailty progression, which included 9.1% and 21.0%, respectively, who progressed to frailty. Predictors in the final nomogram were age, marital status, physical exercise, baseline frailty state, and diabetes. Based on this nomogram, an online calculator was also developed for easy use. The discriminative ability was good in the training set (C index = 0.861) and was validated using both the internal bootstrap method (C index = 0.861) and an external validation set (C index = 0.853). The calibration plots showed good agreement in both the training and validation sets. CONCLUSIONS AND IMPLICATIONS: An easy-to-use nomogram was developed with good apparent performance using 5 readily available variables to help physicians and public health practitioners to identify older adults at high risk for frailty progression and implement medical interventions.

Diagnostic and Predictive Value of Immune-Related Genes in Crohn's Disease.

Abnormal immune cell infiltration is associated with the pathogenesis of Crohn's disease (CD). This study aimed to determine the diagnostic and predictive value of immune-related genes in CD. Seven Gene Expression Omnibus datasets that analyzed the gene expression in CD tissues were downloaded. Single-sample gene set enrichment analysis (ssGSEA) was used to estimate the infiltration of the immune cells in CD tissues. Immune-related genes were screened by overlapping the immune-related genes with differentially expressed genes (DEGs). The protein-protein interaction (PPI) network was used to identify key immune-related DEGs. Diagnostic value of CD and predictive value of anti-TNFα therapy were analyzed. Immunohistochemical (IHC) assay was used to verify gene expression in CD tissues. There were significant differences among CD tissues, paired CD tissues, and normal intestinal tissues regarding the infiltration of immune cells. AQP9, CD27, and HVCN1 were identified as the key genes of the three sub-clusters in the PPI network. AQP9, CD27, and HVCN1 had mild to moderate diagnostic value in CD, and the diagnostic value of AQP9 was better than that of CD27 and HVCN1. AQP9 expression was decreased in CD after patients underwent anti-TNFα therapy, but no obvious changes were observed in non-responders. AQP9 had a moderate predictive value in patients who had undergone treatment. IHC assay confirmed that the expression of AQP9, CD27, and HVCN1 in CD tissues was higher than that in normal intestinal tissues, and AQP9, CD27 was correlated with the activity of CD. Immune-related genes, AQP9, CD27, and HVCN1 may act as auxiliary diagnostic indicators for CD, and AQP9 could serve as a promising predictive indicator in patients who underwent anti-TNF therapy.

Health information needs regarding diabetes mellitus in China: an internet-based analysis.

BACKGROUND: Today,. most people use the Internet to seek online health-related information from general public health-related websites and discussion groups. However, there are no Internet-based analyses of health information needs pertaining to diabetes in China until now. With the development of artificial intelligence,we can analyzed these online health-related information and provide references for health providers to improve their health service. METHODS: We have done a study of statistically analyzing the questions about diabetes collected from 39 health website, the number of which is 151,589. We have divided these questions into 9 categories using a convolutional neural network. RESULTS: The diabetes problems of consumer are presented as follows, diagnosis: 34.95%, treatment: 25.17%, lifestyle: 21.09%, complication: 8.00%, maternity-related:5.00%, prognosis: 2.59%, health provider choosing: 1.40%, prevention: 1.23%, others: 0.58%, The elderly are more concerned about the treatment and complications of diabetes, while the young are more concerned about the maternity-related and prognosis of diabetes. The diabetes drugs most frequently mentioned by consumers are insulin, metformin and Xiaoke pills, The most concerned complication is caidiovascular disease and diabetic eye disease. CONCLUSION: Diabetes health education should focus on how to prevent diabetes and the contents of health education should be different for differernt age groups;on diabetes treatment, the use of insulin and oral hypoglycemic drugs education should be strengthened.

No association of G-protein beta polypeptide 3 polymorphism with irritable bowel syndrome: evidence from a meta-analysis.

AIM: To clarify the associations between G-protein beta polypeptide 3 (GNB3) C825T polymorphism and risk of the irritable bowel syndrome (IBS) by a meta-analysis. METHODS: We searched relevant studies in PubMed, EMBASE, CNKI, Google Scholar, Ovid and Cochrane library prior to October 2013. The strengths of the associations between GNB3 C825T polymorphism and IBS risk were estimated by odds ratios (ORs) with 95% confidence interval (CIs). RESULTS: We identified seven case-control studies with 1085 IBS cases and 1695 controls for the analysis. We found no significantly associations of GNB3 C825T polymorphism with IBS risk in the overall population (CC vs TT, OR = 1.12, 95%CI: 0.86-1.45; CC + CT vs TT, OR = 1.17, 95%CI: 0.92-1.49; TT + CT vs CC, OR = 0.93, 95%CI: 0.80-1.08; C vs T, OR = 1.08, 95%CI: 0.97-1.21). Subgroup analysis did not reveal significant associations either in Asian population or Caucasian population. The pooled results of four studies fail to show associations of GNB3 C825T polymorphism with subtypes of IBS (constipation-dominant type, diarrhea-dominant type and mixed type). CONCLUSION: The present study suggests no associations of GNB3 C825T polymorphism with IBS risk.

Accelerated healing of diabetic wound using artificial dermis constructed with adipose stem cells and poly (L-glutamic acid)/chitosan scaffold.

BACKGROUND: Diabetic wound is one of the most serious complications of diabetes mellitus. There are no significantly effective therapies for chronic non-healing diabetes ulcer so far. This study aimed to explore the feasibility of healing impaired wound using artificial dermis constructed with human adipose derived stem cells (ASCs) and poly (L-glutamic acid)/chitosan (PLGA/CS) scaffold in streptozotocin-induced diabetic mice. METHODS: ASCs were isolated from fresh human lipoaspirates and expanded ex vivo for three passages, and then cells were seeded onto PLGA/CS scaffold to form artificial dermis. Expression of VEGF and TGFß1 by ASCs presented in artificial dermis was determined. The artificial dermis was transplanted to treat the 20 mm × 20 mm full-thickness cutaneous wound created on the back of diabetic mice. Wound treated with scaffold alone and without treatment, and wound in normal non-diabetic mice served as control. RESULTS: Cells growing within scaffold showed great proliferation potential, depositing abundant collagen matrix. Meanwhile, expression of VEGF and TGF-ß1 by seeded ASCs maintained at a consistent high level. After treated with ASC based artificial dermis, diabetic wounds exhibited significantly higher healing rate compared with wounds treated with scaffold alone or without treatment. Histological examination also demonstrated an improvement in cutaneous restoration with matrix deposition and organization. Further quantitative analysis showed that there was a significant increase in dermis thickness and collagen content on artificial dermis treated wounds. CONCLUSION: ASC/PLGA artificial dermis can effectively accelerate diabetic wound healing by promoting angiogenic growth factors and dermal collagen synthesis.

Ubiquinone-quantum dot bioconjugates for in vitro and intracellular complex I sensing.

Quantum dots (QDs) have attracted increasing interest in bioimaging and sensing. Here, we report a biosensor of complex I using ubiquinone-terminated disulphides with different alkyl spacers (QnNS, n = 2, 5 and 10) as surface-capping ligands to functionalise CdSe/ZnS QDs. The enhancement or quenching of the QD bioconjugates fluorescence changes as a function of the redox state of QnNS, since QDs are highly sensitive to the electron-transfer processes. The bioconjugated QnNS-QDs emission could be modulated by complex I in the presence of NADH, which simulates an electron-transfer system part of the mitochondrial respiratory chain, providing an in vitro and intracellular complex I sensor. Epidemiological studies suggest that Parkinson's patients have the impaired activity of complex I in the electron-transfer chain of mitochondria. We have demonstrated that the QnNS-QDs system could aid in early stage Parkinson's disease diagnosis and progression monitoring by following different complex I levels in SH-SY5Y cells.

Surfactant-free microwave-assisted hydrothermal synthesis of BaMoO4 hierarchical self-assemblies and enhanced photoluminescence properties.

BaMoO(4) with 3D hierarchical multilayer disk-like and nest-like architectures self-assembled from 2D nanosheets was successfully synthesized via a microwave-assisted hydrothermal route without any surfactant. The as-prepared products were characterized by X-ray powder diffractometer (XRD), scanning electron microscope (SEM), field emission transmission electron microscope (FE-TEM), and photoluminescence (PL) spectrometer. The results show that the reaction parameters, including pH value, reactant concentration, and molar ratio of [Ba(2+)]/[MoO(4)(2-)], played important roles on the morphologies of the final products. And the formation mechanism of 3D hierarchical architectures is a stepwise oriented aggregation-based self-assembly process. The superstructure characteristic of 3D nest-like BaMoO(4) architecture was observed in HRTEM image and the corresponding fast Fourier transform (FFT) for the first time, and the superlattice reflections with non-integer indices occurred around the subcell reflections at ±1/6(2a*+2c*). Room temperature photoluminescence spectra of 3D BaMoO(4) architectures reveal a strong and broad blue emission, and the 3D nest-like architectures own the enhanced intensity than multilayer disks.

Sigmoid colonic carcinoma associated with deposited ova of Schistosoma japonicum: a case report.

We report a case of sigmoid colonic carcinoma associated with deposited ova of Schistosoma japonicum. A 57-year old woman presented with a 10-mo history of left lower quadrant abdominal pain and a 2-mo history of bloody stools. She had a significant past medical history of asymptomatic schistosomiasis japonica and constipation. A colonoscopy showed an exophytic fragile neoplasm with an ulcerating surface in the sigmoid colon. During the radical operative procedure, we noted the partially encircling tumor was located in the distal sigmoid colon, and extended into the serosa. Succeeding pathological analysis demonstrated the diagnosis of sigmoid colonic ulcerative tubular adenocarcinoma, and showed deposited ova of Schistosoma japonicum in both tumor lesions and mesenteric lymph nodes. Three days after surgery the patient returned to the normal bowel function with one defecation per day. These findings reveal that deposited schistosome ova play a possible role in the carcinogenesis of colorectal cancer.

Hypoxia can contribute to the induction of the Epstein-Barr virus (EBV) lytic cycle.

BACKGROUND: Like other herpes viruses, latent Epstein-Barr virus (EBV) infection can be reactivated to lytic replication. Reactivation can be achieved by treatment with various reagents, including tetradecanoyl phorbol acetate (TPA) and Ca2+ ionophores. Relatively little is known about the physiological factors related to reactivation of EBV. Previous studies have demonstrated that G0/G1 cell cycle arrest is associated with EBV activation, and that hypoxic conditions can induce cell cycle arrest. In the present study we investigated the effect of hypoxia on reactivation of EBV. OBJECTIVE AND METHODS: Hypoxic culture conditions were established and the expression of Zta protein and the number of EBV DNA copies were measured in B95-8 cells maintained under these conditions. RESULTS: Hypoxia treatment not only increased the expression of the EBV immediate-early protein Zta (which mediates the switch between the latent and lytic form of infection), but also increased the number of EBV DNA copies in B95-8 cells. CONCLUSIONS: EBV in latent infection can be activated to lytic infection by hypoxia treatment.

The expression of p63 is associated with the differential stage in nasopharyngeal carcinoma and EBV infection.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is common among Southern Chinese and the main histology is the undifferentiated carcinoma associated with Epstein-Barr virus (EBV) infection. p63 is a recently proved member of the p53 family based on the structural similarity to p53, but its function in NPC is still unknown. This study was aimed to investigate the association between p63 and NPC. RESULTS: p63 was expressed in 100% (202/202) of nasopharyngeal carcinoma (NPC) tissues but not in 29 nasopharynx inflammation and 17 non-cancerous nasopharyngeal epidermises on a tissue microarray by immunohistostaining. Further investigation suggested that the p63 expression was associated with the differential stage of NPC: p63 strong staining in Keratinizing squamous cell carcinoma, differentiated non-keratinizing NPC and undifferentiated non-keratinizing NPC presented the percentage of 5/8 (62.5%), 43/48 (92.5%) and 50/50 (100%), respectively. A significant difference (p = 0.001) existed between the keratinizing and non-keratinizing groups. No pathogenic mutations were detected in p63 gene in 12 primary NPC tissues and matched peripheral blood lymphocytes (PBL). Half-life measurement study revealed distinct stability of p63 protein in the different cell lines, especially between the carcinoma cell lines with EBV infection and the non-cancerous cell lines. The results of immunoprecipitation suggested a direct interaction between Epstein-Barr virus nuclear antigen 5 (EBNA-5) and p63 protein in NPC, and this binding would increase the stability of p63. CONCLUSION: Our data suggested p63 might be used as an adjunct diagnostic marker of NPC and contributed a new way to understand the contribution of the EBV in the pathogenesis of NPC.

High frequency somatic mutations in RASSF1A in nasopharyngeal carcinoma.

High frequency loss of 3p21.3 region is a common event in various kinds of tumors including nasopharyngeal carcinoma (NPC). RASSF1A has been identified as a putative tumor suppressor gene residing in this region. Chromosome alterations and epigenetic changes are commonly observed as mechanisms for inactivation of RASSF1A function. In this study, we applied the PCR-cloning-sequencing strategy to examine somatic mutations in RASSF1A in NPC tissues as compared with the sequences detected in the matched peripheral blood lymphocytes. Our results revealed a high incidence of RASSF1A mutation in primary tumor tissues of NPC. There are totally 35 mutations identified in 74% (17/23) of these NPC cases, including 30 transitions, three transversions and two deletions. Most of these mutations result in amino acid changes: three nonsense (stop codon) mutations, two-1 bp deletion (frameshift), 26 missense and the remaining four are synonymous (silent). No obvious 'hot-spot' mutations were observed in this study. A similarly high rate (74%) of promoter methylation of RASSF1A was also detected in the same group of NPC tissues, but no significant correlation between mutation and methylation was detected. Our results suggest various mechanisms involved in inactivation of RASSF1A function and indicate a critical role of RASSF1A in NPC development.

Detection of herpes simplex virus type 1 in rheumatic valvular tissue.

BACKGROUND: Rheumatic heart disease (RHD) is the most important sequela of rheumatic fever (RF): evidence that streptococcal infection is aetiological is prominent, but sometimes contradictory. Acute HSV-1 infection in mouse leads to carditis and valvulitis whereas recurrent infection results in inflammatory granulomatous lesions that resemble Aschoff bodies. Cells containing HSV-1 inclusions or virus infected giant cells appear similar to Anitschkow cells or Aschoff cells respectively. We hypothesized that HSV-1 infection also may be involved in RHD. METHODS: Formalin-fixed, paraffin-embedded valvular tissue samples from 32 patients with RHD were investigated for evidence of HSV-1 infection. HSV-1 antigen was detected by immunohistochemistry, using HSV-1-specific monoclonal and polyclonal antibodies. HSV-1 glycoprotein D gene sequences were amplified by nPCR, using beta-globin gene amplification in the same samples as internal control. Valvular tissue from 5 cases of sudden death and 3 cases died of neisseria meningitis without a history of valvular disease was used for comparison. HSV-1-infected lung tissue was used as positive control. RESULTS: HSV-1 antigens were detected in valvular tissues from 21 of 32 (65.6%) patients. Fifteen of these 21 (46.9% of cases), but no antigen-negative sample, were positive also for HSV DNA. Nucleotide sequence of PCR products was homologous to the targeted region of the HSV-1 glycoprotein D gene. HSV-1 antigen was present also in one case of sudden death but viral DNA was not found in any tissue sample from the comparison group. Results from reagent and positive controls were as anticipated. CONCLUSIONS: This is the first study to show the presence of HSV-1 antigen and genomic DNA in valvular tissues from patients with RHD and provides evidence for an association of HSV-1 infection with some cases of rheumatic valvular disease.

Transcriptional expression of RPMS1 in nasopharyngeal carcinoma and its oncogenic potential.

The Epstein-Barr virus (EBV) BamHI A rightward transcripts (BARTs) were originally identified in C15 xenograft of nasopharyngeal carcinoma (NPC) and easily detected in a wide variety of EBV latent infection and EBV-associated tumors. It had been reported that p31 cosmid containing BARTs immortalized monkey epithelial cells, but which particular gene among BARTs family participates in the transformation procedure remains to be identified. RPMS1 is the only full-length cDNA confirmed so far and one of the most abundant spliced forms in BARTs family. To investigate the involvement of RPMS1 gene in NPC, we examined the expression of RPMS1 transcripts in NPC biopsies from Guangdong and its oncogenic potential. Our results revealed that RPMS1 mRNA preferentially expressed in primary NPC to non-carcinoma tissue of nasopharynx and peripheral blood lymphocytes (PBLs) of NPC patients. Furthermore, by introducing RPMS1 ORF into HEK293 cells, these transfectants enhanced the anchorage-independent growth and produced tumors in nude mice. These data imply that RPMS1 gene might play an important role in the development of NPC.

Transcriptional gene expression profile of human nasopharynx.

Inflammation in the nasopharynx is very common worldwide and nasopharyngeal carcinoma (NPC) results in significant morbidity and mortality in southeast Asia and north Africa. To facilitate the understanding of pathogenesis of these diseases as well as normal nasopharynx biology, transcriptional gene expression profile of normal human nasopharynx, from undissected biopsies, was established in this study by generating a large amount of ESTs, followed by bioinformatics analysis. A total of 27,209 ESTs generated from human nasopharynx cDNA library were integrated into 8,420 non-redundant clusters, of which, 6,070 (72.09%) corresponded to known genes, 156 (1.85%) to known ESTs, and 2,194 (26.06%) to novel sequences, respectively. Functional classification revealed that transcripts constituting enzymes (2,284, 28.30%) and those participating in cell growth/maintenance (3,306, 46.33%) were the largest population expressed in the nasopharynx, followed by genes coding for binding proteins (2,135, 26.45%) and involved in cell communication process (1,832, 25.68%). In addition, through comparison of the nasopharynx gene expression profile with those of 7 other human tissues, 59 transcripts preferentially expressed and 22 transcripts unique in the nasopharynx were identified. Our study acquired an initial assessment of qualitative diversity of gene expression in the nasopharynx, providing the first step toward comprehensive characterization of the molecular features of the nasopharyngeal disorders.

[Expression of cytochrome P450 enzymes in human nasopharyngeal carcinoma and non-cancerous nasopharynx tissue].

BACKGROUND & OBJECTIVE: The etiology of nasopharyngeal carcinoma (NPC) is associated with environmental and hereditary factors. Xenobiotics from environment must be activated to derive carcinogens. Several cytochrome P450 (CYP450) metabolic enzymes participate to the activation of pre-carcinogens, and the genetic polymorphism of those genes is associated with metabolic polymorphism and susceptibility to cancer. We performed a preliminary investigation on the xenobiotics metabolism of human nasopharynx by determining the expression of CYP450 genes in NPC and non-cancerous nasopharynx tissues. METHODS: The following two methods were used: (1)A cDNA library from the mixed RNA sample of seven non-cancerous nasopharynx tissues was generated, followed by clone sequencing and bioinformatics analysis. (2)RNA of 14 NPC and 8 non-cancerous nasopharynx tissues were reversely transcribed, and the expression of CYP450 genes in those samples was determined by PCR amplification. RESULTS: Eight ESTs of CYP450 genes including CYP1B1, CYP2F1, CYP2J2, CYP4B1, CYP4F12, CYP5A(TBXAS1), CYP20A1, and CYP51A1 were detected in non-cancerous nasopharynx cDNA library, among these CYP4B1 exhibited the highest expression level with 16 copies of ESTs. Positive expression of fourteen CYP450 genes including CYP1A1, CYP1B1, CYP2A6, CYP2A13, CYP2B6, CYP2C8, CYP2C9, CYP2D6, CYP2E1, CYP2F1, CYP3A4, CYP3A5, CYP3A7, and CYP4B1 were detected by RT-PCR, among these, CYP1B1, CYP2B6, and CYP4B1 had also been detected in the cDNA library. A total of 19 CYP450 genes expression were detected in NPC and non-cancerous nasopharynx tissues, and the expression levels of CYP1B1, CYP2C8, CYP2C9, CYP2D6, CYP3A5, and CYP4B1 were higher than those of the other genes. CONCLUSION: The expression of a great number of CYP450 genes was detected in human nasopharynx, some of which might participate to the activation of pre-carcinogen in human nasopharynx. Contribution of these genes to the risk of NPC needs further investigation.

Genetic polymorphisms of CYP2A13 and its relationship to nasopharyngeal carcinoma in the Cantonese population.

Nasopharyngeal carcinoma (NPC) is characterized by a high prevalence in Southern China, especially among Cantonese individuals of the Guangdong Province. Epidemiological studies have suggested that frequent exposure to high levels of nitrosamine from preserved foods such as salted fish could be a risk factor for NPC. Cytochrome P450 encompasses a family of enzymes that metabolize carcinogens and CYP2A13, a member of this family, is expressed predominantly in the respiratory tract with the highest levels in the nasal mucosa. In an effort to test whether a correlation exists between CYP2A13 genetic polymorphism and the risk of developing NPC, we sequenced all nine exons and the exon-intron junctions of the CYP2A13 gene in 45 NPC patients. We identified a total of 21 single nucleotide polymorphism (SNPs), including 7 novel SNPs. The most frequent functional variant allele was 74A-1757G-3375T-7233G with a haplotype frequency of 7.8% in the 45 NPC cases. In addition, a stop codon mutation was detected in one case. We then selected the 3 most frequent SNPs and one stop codon mutation to expand our study to a case-control analysis within the Cantonese population. A novel haplotype consisting 8 SNPs in introns, and four additional novel SNPs were identified; but no correlation between CYP2A13 genetic polymorphism and individual susceptibility to NPC was observed.

Alterations of BLU, a candidate tumor suppressor gene on chromosome 3p21.3, in human nasopharyngeal carcinoma.

Nonrandom allelic loss on chromosome 3p is a common event in nasopharyngeal carcinoma (NPC) with the implication that certain tumor suppressor gene(s) in this region are involved in the pathogenesis of these tumors. The BLU gene, located at 3p21.3, has recently been identified as a candidate tumor suppressor gene due to the occurrence of missense mutations and loss of its expression in lung cancer. To investigate the involvement of BLU gene in NPC, we examined both genetic and epigenetic changes of BLU in NPC primary tumors and cell lines. No pathogenic mutations were detected in the entire coding region of this gene in 45 primary NPC tumors and 5 NPC cell lines. While BLU was expressed in 100% (15 of 15) of noncancerous nasopharyngeal epithelia, its transcripts were missing in all 5 NPC cell lines, and absent or reduced mRNA levels were observed in 78% (28 of 36) of the primary tumors. In the NPC cell lines, loss of BLU expression correlated with hypermethylation of the CpG island promoter sequence, and expression was restored after treatment with 5'-aza-2'-deoxycytidine. Methylation specific PCR analysis revealed that the BLU promoter was highly methylated in 74% (17 of 23) of primary tumors in which BLU was downregulated, whereas only 2 of 9 non-neoplastic nasopharyngeal epithelia exhibited hypermethylation in the BLU promoter region. The high incidence of BLU alterations suggests that it may be one of the critical tumor suppressor genes on chromosome 3p21.3 involved in the development of NPC.

[Mutation and expression of SEMA3B and SEMA3F gene in nasopharyngeal carcinoma].

BACKGROUND & OBJECTIVE: Though the molecular etiology of nasopharyngeal carcinoma(NPC) is currently unknown, evidence from both loss of heterozygosity analysis and functional studies suggested that there are NPC-associated tumor suppressor genes(TSGs) residing in chromosome 3p21.3. Recently, two members of semaphorin family, SEMA3B and SEMA3F gene, located at 3p21.3, were characterized as TSGs. Studies showed that SEMA3B and SEMA3F are capable of suppressing the growth of tumor cells and inducing apoptosis. Loss of SEMA3B mRNA expression or aberrant SEMA3F cellular localization were found in lung cancers. In order to investigate the involvement of SEMA3B and SEMA3F in NPC, the authors examined both mutation and expression of these two genes in NPC. METHODS: The entire coding regions, the splice donor/acceptor sites, and partial regulatory regions of SEMA3B and SEMA3F gene were screened for mutations by PCR-sequencing in 21 primary NPC tumors and 2 NPC cell lines(CNE2 and SUNE1). The mRNA expression levels were determined by semi-quantitative RT-PCR analysis. RESULTS: No somatic mutation was found in either SEMA3B or SEMA3F gene. However, two missense polymorphisms including Thr415Ile and lle242Met were found in SEMA3B in NPC. For the Thr415Ile polymorphism, the Ile allele type which leads to SEMA3B function defects was predominant in NPC with the allele frequency of 64% (27/42). SEMA3B mRNA was expressed in all 6 non-neoplastic nasopharyngeal epithelia, but was absent or down-regulated in 76% (16/21) of primary NPC tumors. No significant difference of SEMA3B expression was observed between NPC and noncancerous controls. CONCLUSION: High frequency of SEMA3B expression alterations suggests that the inactivation of this gene was strongly associated with NPC. SEMA3B may be a tumor suppressor on 3p21.3 involved in NPC.

[Analysis of suppressive roles of BLU gene at 3p21.3 in nasopharyngeal carcinoma].

BACKGROUND & OBJECTIVE: Nonrandom allelic loss at chromosome 3p21.3 is a common and early event in nasopharyngeal carcinoma (NPC), which implicates the presence of tumor suppressor genes (TSGs) that may be involved in the pathogenesis of NPCs. BLU gene, containing a MYND domain and located at 3p21.3, has been considered as a NPC associated candidate tumor suppressor gene (TSG) due to the occurrence of loss of its expression and aberrant promoter hypermethylation in most NPCs. This study was designed to construct expression vectors containing either wild type BLU gene and its mutants and to analyze the effect of BLU gene on proliferation of NPC cells by transfection assays. METHODS: The full-length cDNA of BLU gene was amplified by RT-PCR. The expression vectors containing various BLU mutants were constructed by site-directed mutagenesis by overlapping PCR. These mutants include a MYND domain deletion mutant, a Ser402Phe and del405Cys, del406Ser mutant, and a Gly160Arg mutant. The wild type BLU gene and the MYND domain deletion mutant were transfected into NPC cell lines CNE1 and CNE2. The effect on apoptosis was determined by TUNEL assay. Cellular proliferation of the stably-transfected cells was examined with cell growth curve and by colony formation assays. Tumorigenicity in nude mice of CNE2 stably-transfected with BLU was investigated. RESULTS: No significant difference in apoptosis index (AI) was observed between cells transfected with wild type or MYND domain deleted BLU gene and cells transfected with plasmid controls. Exogenous expression of wild type BLU gene had no effect on growth rate and colony formation ability of CNE1 and CNE2. BLU gene showed no suppressor ability in CNE2 tumorigenicity. CONCLUSION: Although BLU gene was frequently altered in NPCs, its suppressor role in NPC cells proliferation was not evident. Thus, the possibility of BLU gene as a TSG involved in NPC development remained to be elucidated by further studies.