DNA-PKcs participated in hypoxic pulmonary hypertension.

BACKGROUND: Hypoxic pulmonary hypertension (HPH) is a common complication of chronic lung disease, which severely affects the survival and prognosis of patients. Several recent reports have shown that DNA damage and repair plays a crucial role in pathogenesis of pulmonary arterial hypertension. DNA-dependent protein kinase catalytic subunit (DNA-PKcs) as a part of DNA-PK is a molecular sensor for DNA damage that enhances DSB repair. This study aimed to demonstrate the expression and potential mechanism of DNA-PKcs on the pathogenesis of HPH. METHODS: Levels of DNA-PKcs and other proteins in explants of human and rats pulmonary artery from lung tissues and pulmonary artery smooth muscle cells (PASMC) were measured by immunohistochemistry and western blot analysis. The mRNA expression levels of DNA-PKcs and NOR1 in PASMCs were quantified with qRT-PCR. Meanwhile, the interaction among proteins were detected by Co-immunoprecipitation (Co-IP) assays. Cell proliferation and apoptosis was assessed by cell counting kit-8 assay(CCK-8), EdU incorporation and flow cytometry. Rat models of HPH were constructed to verify the role of DNA-PKcs in pulmonary vascular remodeling in vivo. RESULTS: DNA-PKcs protein levels were both significantly up-regulated in explants of pulmonary artery from HPH models and lung tissues of patients with hypoxemia. In human PASMCs, hypoxia up-regulated DNA-PKcs in a time-dependent manner. Downregulation of DNA-PKcs by targeted siRNA or small-molecule inhibitor NU7026 both induced cell proliferation inhibition and cell cycle arrest. DNA-PKcs affected proliferation by regulating NOR1 protein synthesis followed by the expression of cyclin D1. Co-immunoprecipitation of NOR1 with DNA-PKcs was severely increased in hypoxia. Meanwhile, hypoxia promoted G2 + S phase, whereas the down-regulation of DNA-PKcs and NOR1 attenuated the effects of hypoxia. In vivo, inhibition of DNA-PKcs reverses hypoxic pulmonary vascular remodeling and prevented HPH. CONCLUSIONS: Our study indicated the potential mechanism of DNA-PKcs in the development of HPH. It might provide insights into new therapeutic targets for pulmonary vascular remodeling and pulmonary hypertension.

Clinical features of Epstein-Barr virus in the intestinal mucosa and blood of patients with inflammatory bowel disease.

BACKGROUND: The role of Epstein-Barr virus (EBV) in inflammatory bowel disease (IBD) remains to be elucidated. The aim of this study was to investigate the presence of EBV in the blood and intestinal mucosa of patients with IBD and evaluate the association between EBV positivity and IBD. METHODS: Patients with IBD, hospitalized between January 2015 and April 2018, were enrolled. The EBV-DNA load in blood samples from each subject was analyzed by quantitative real-time polymerase chain reaction. EBV-encoded small-RNA 1 (EBER-1) was detected by in-situ hybridization in intestinal mucosa tissue sections of patients with IBD. RESULT: EBV-DNA was detected in 48 out of 568 patients with IBD (8.4%), and EBER-1 positivity was detected in 27 of these patients (56.3%). Refractory IBD and severe mucosal inflammation were more common in patients with detectable levels of EBER-1 than in those without; the number of EBER-1-positive cells positively correlated with mucosal inflammation (P value < 0.05). Age (≥60 years old) and use of azathioprine were risk factors for EBV infection. There was no significant difference in clinical remission rate and surgical rate between the EBER-1 positive group and EBER-1 negative group, antiviral group and the non-antiviral group, among IBD patients who tested positive for EBV-DNA. CONCLUSION: Elderly patients with IBD, treated with azathioprine, are more susceptible to EBV positivity. Further, EBV mucosal detection correlated with the severity of mucosal damage and refractoriness, but not prognosis.

[Short-Term Therapeutic Efficacy, Survival Time and Side Effects in Treatment of Newly Diagnosed Multiple Myeloma Patients with Different Chemotherapeutic Regimens].

OBJECTIVE: To investigate the short-term therapeutic efficacy, survival time and side effects in newly diagnosed multiple myeloma patients treated with TCD regimen consisted of thalidomide, cyclophosphiamide and dexamethasone, and BCD reginen consisted of bortezomib, cyclophsphamide and dexamethasone. METHODS: The clinical data of newly diagnosed MM patients admitted in our hospital from January 2011 to January 2018 were collected and analyzed retrospectively. According to chemotherapectic regimen, 106 patients were divided into 2 groups: 53 cases in one group were treated with TCD regimen (TCD group), and 53 cases in another group were treaded with BCD regimen (BCD group). The therapeutic efficacy, median PFS and OS time and incidence of side effects in 2 groups were compared, at the same time the relationship of the remission degree and the factors in different subgroups with the therapeutic efficacy was analyzed in 2 groups. RESULTS: There was no significant difference in the ≥MR rate between 2 groups (P＞0.05). The ≥PR rate, ≥VGPR rate and CR rate of BCD group were significantly higher than TCD group (P＜0.05). The median PFS time of patients in BCD group were significantly longer than that in TCD group (P＜0.05). There was no significant difference in the median OS time of patients between 2 groups (P＜0.05). The median OS time of ≥MR patients in TCD group was significantly longer than that of ＜MR patients (P＜0.05). The median OS time of ≥PR patients in TCD group were significantly longer than that of ＜PR patients (P＜0.05). The median OS time of ≥VGPR patients in BCD group was significantly longer than that of ＜VGPR patients (P＜0.05). There was no significant difference in the median OS time of ≥PR and ＜PR patients in BCD group (P＞0.05). The ORR of ≥VGPR patients in BCD group was significantly higher than that in TCD group (P＜0.05). There was no significant difference in the incidence of infection, fatigue, gastrointestinal reactions and bone marrow suppression between 2 groups (P＜0.05). The incidence of numbness in distal extremities and herpes zoster in BCD group was significantly higher than that in TCD group (P＜0.05). CONCLUSION: TCD and BCD in the treatment of patients with NDMM possess the same disease control effects; BCD regimen application can efficiently improve remission degree and prolong PFS time; but TCD regimen application have the advantages in reducing the side effects risk and improving treatment tolerance.

Clinicopathologic characteristics of HER2-positive pure mucinous breast carcinoma: a systematic investigation into an unusual tumor.

Pure mucinous breast carcinoma (PMBC) accounts for approximately 2% of all breast carcinoma. Overexpression or amplification of human epidermal growth factor receptor 2 (HER2) is rarely observed in PMBC. We retrieved 119 PMBCs, which included 12 HER2-positive PMBCs and 107 HER2-negative PMBCs, to compare the clinicopathologic features between HER2-positive and HER2-negative neoplasms. The assessed parameters included patient age, menstruation, laterality, tumor size, lymph node status, tumor-node-metastasis (TNM) stage, nuclear grade, receptor status, treatment and prognostic features. HER2-positive PMBCs represented approximately 10.1% of the PMBCs examined. HER2-positive PMBCs showed more frequent lymph node metastasis (P=0.038), a significantly higher clinical TNM stage (P<0.001) and nuclear grade (P<0.001), lower estrogen receptor (ER) and progesterone receptor (PR) expression and higher Ki67 expression than the HER2-negative group (P=0.011, P=0.005, and P=0.001, respectively). HER2-positive PMBCs (untreated with HER2-targeted therapy) had a significantly lower overall survival (OS) rate than HER2-negative PMBCs (P=0.005). Nodal metastasis, higher TNM stage and nuclear grade were identified as factors that result in poorer OS of patients with PMBCs (P<0.001, P=0.016, P<0.001, and P<0.001, respectively). Univariate and multivariate Cox analyses confirmed that HER2 status was an independent prognostic factor for PMBCs (P=0.003 and P=0.012, respectively). HER2-positive PMBC is a rare subtype of breast carcinoma with aggressive biological behavior. It is important to identify tumors with these aggressive clinical behaviors and manage them differently. To the best of our knowledge, this study represents the first systematic investigation of the clinicopathologic features of HER2-positive PMBCs.

Clinicopathological and molecular characteristics of synchronous gastric adenocarcinoma and gastrointestinal stromal tumors.

Synchronous gastric tumors that consist of both gastrointestinal stromal tumor (GIST) and adenocarcinoma are rare. We studied the clinicopathological and molecular characteristics of six cases containing both gastric adenocarcinoma and GIST. By means of immunohistochemical analysis, all GIST cells expressed CD117, CD34 and Dog1 in all six synchronous gastric adenocarcinomas with GIST, and in GIST alone. Sequencing analysis demonstrated that exon 11 c-kit mutations were present in two of six synchronous tumors and four of five GISTs. One of the two exon 11 c-kit mutations in synchronous adenocarcinomas with GISTs was an uncommon mutation of CTT > CCA at amino acid 576, and the other was a GTT deletion at amino acid 560. The mutation was a homozygous A > G mutation in exon 12 (amino acid 567) of PDGFR-α. We concluded that the exon 11 mutations were the most important in both cases of synchronous gastric adenocarcinoma with GIST and GIST alone. The mutation rate was higher in GIST alone than in synchronous adenocarcinoma with GIST.

[Study on reduced folate carrier gene (RFC1) polymorphism in the southern and northern Chinese population].

OBJECTIVE: To describe the distribution of reduced folate carrier gene (RFC1)genotype and allele frequency between southern and northern, female and male Chinese population. METHOD: RFC1 (A80G) genotype was detected, using polymerase chain reaction-restriction fragment length polymorphism (RFLP-PCR) on 720 blood spot DNA from the normal subjects. RESULTS: The frequencies of the northern population with AA, GG and GA genotypes were 22.28%, 31.09% and 46.63%, and the frequencies of the southern population were 18.56%, 22.75% and 58.68%, respectively. Findings showed that there were significant differences between southerners and northerners in RFC1 (A80G) genotype (P < 0.01). There was no significant difference between G allele frequency of the northern (52.10%) and southern population (54.40%). The frequencies of male with RFC1 (A80G) AA, GG and GA genotype were 24.88%, 25.85% and 49.27%, and among female were 18.83%, 27.77% and 53.40%, respectively. There were no significant differences between male and female in RFC1 genotype (P > 0.05), or between G allele frequency in female (50.49%) and that in male (54.47%). CONCLUSIONS: The distribution of RFC1 genotype seemed to be consistent with neural tube defects (NTDs) while its prevalence among the northerners was higher than that of southerners, with female having a higher NTDs prevalence. This study provided genetic epidemiological data for etiological hypothesis between RFC1 and diseases relative to folate metabolism.