Microscopic Mechanism of Proton Transfer in Pure Water under Ambient Conditions.

Water molecules and the associated proton transfer (PT) are prevalent in chemical and biological systems and have been a hot research topic. Spectroscopic characterization and ab initio molecular dynamics (AIMD) simulations have previously revealed insights into acidic and basic liquids. Presumably, the situation in the acidic/basic solution is not necessarily the same as in pure water; in addition, the autoionization constant for water is only 10-14 under ambient conditions, making the study of PT in pure water challenging. To overcome this issue, we modeled periodic water box systems containing 1000 molecules for tens of nanoseconds based on a neural network potential (NNP) with quantum mechanical accuracy. The NNP was generated by training a dataset containing the energies and atomic forces of 17 075 configurations of periodic water box systems, and these data points were calculated at the MP2 level that considers electron correlation effects. We found that the size of the system and the duration of the simulation have a significant impact on the convergence of the results. With these factors considered, our simulations showed that hydronium (H3O+) and hydroxide (OH-) ions in water have distinct hydration structures, thermodynamic and kinetic properties, e.g., the longer-lasting and more stable hydrated structure of OH- ions than that of H3O+, as well as a significantly higher free energy barrier for the OH--associated PT than that of H3O+, leading the two to exhibit completely different PT behaviors. Given these characteristics, we further found that PT via OH- ions tends not to occur multiple times or between many molecules. In contrast, PT via H3O+ can synergistically occur among multiple molecules and prefers to adopt a cyclic pattern among three water molecules, while it occurs mostly in a chain pattern when more water molecules are involved. Therefore, our studies provide a detailed and solid microscopic explanation for the PT process in pure water.

Awareness of and Preferences for Preexposure Prophylaxis (PrEP) among MSM at High Risk of HIV Infection in Southern China: Findings from the T2T Study.

This study is aimed at investigating the awareness of and preferences for oral and long-acting injectable HIV preexposure prophylaxis (PrEP) and their associated factors among men who have sex with men (MSM) at high risk of HIV infection in southern China. A cross-sectional survey was conducted among 603 MSM who were recruited through a cohort study called the T2T Study at three sexual health clinics in Guangzhou, Shenzhen, and Wuxi, China, from 2017 to 2018. We collected information on HIV-negative participants' awareness of and willingness to use PrEP and its potential correlations. Univariate and multivariate logistic regressions were used for data analyses. A total of 550 HIV-negative MSM were enrolled in the study. Less than half of at-risk MSM (43.1%) had heard of PrEP before, and the rate of overall willingness to use PrEP was 65.8%, while MSM were more willing to use daily oral PrEP than long-acting injectable- (LAI-) PrEP (62.2% vs. 38.5%). MSM who had university degrees or above (aOR = 1.55, 95% CI: 1.01-2.37), used condoms during last anal sex (1.52, 1.01-2.29), and tested 3 times or more for HIV (2.45, 1.10-5.47) were more likely to be aware of PrEP. MSM who had use of gay dating apps (1.51, 1.02-2.23), ever participated in HIV- or sexually transmitted disease (STD-) related studies (1.91, 1.24-2.94), and had heard of PrEP (3.06, 2.06-4.54) were more willing to use any regimen of PrEP. MSM at high risk of HIV infection had low awareness of PrEP and moderate willingness to use PrEP. Further studies of the implementation and promotion of PrEP targeting at-risk MSM should be performed.

Maternal vitamin D intake during pregnancy and risk of asthma and wheeze in children: a systematic review and meta-analysis of observational studies.

Aims: To summarize the relationship between vitamin D and infant asthma or wheeze.Materials and methods: We used PubMed and Embase to search articles through July 2017 with selection criteria for relevant studies. Random-effect models were used to pool the results of included studies.Results: Ten articles with 14 independent reports of 2073 incident cases of asthma and 1875 cases of wheeze among 23 030 pairs of mother and child were included in our meta-analysis. Compared to those who did not take vitamin D, the mothers who had vitamin D supplementation during pregnancy stage could reduce the risk of asthma or wheeze in infants. The combined odds ratio of infant wheeze was 0.65 (95% CI = 0.54-0.79) and asthma was 0.78 (95% CI = 0.69-0.89). The results almost did not change in the subgroup analyses.Conclusions: It suggests that increasing maternal vitamin D intake during pregnancy might have a protective effect on suffering from wheeze and asthma for children.

Micromechanism Study of Strengthening Effect of Copper on Gold Thiosulfate Leaching.

To study the effect of copper on gold thiosulfate leaching, the gold dissolution of three different sample powders (gold, gold/copper, and gold/copper oxide) in a solution of 5 mM Cu2+, 0.4 M ammonia, and 0.1 M thiosulfate was studied. Scanning electron microscopy analysis showed no sulfur passivation on the gold surface, and there were more prominent corrosion pits on the gold surfaces of samples that were ground with copper or copper oxide. The Evans diagrams showed that copper and copper oxide can promote both the anode and cathode processes of gold dissolution. Based on first principle simulations, copper and copper oxide exhibited the ability to disrupt the stability of gold surface atoms and cause different degrees of relaxation. Both copper and copper oxide reduce the d-band center of the gold surface atoms and the adsorption between gold and thiosulfate. In addition, the bond length of the S-S bond of thiosulfate adsorbed onto the gold surface was longer when copper or copper oxide were not present. According to the change in the potential surface energy, the energy barriers for gold atom dissolution from gold, gold/copper, and gold/copper oxide surfaces were 1.79, 0.72, and 1.01 eV, respectively.

Oral microbial community analysis of the patients in the progression of liver cancer.

Liver disease has been reported to associate with oral microbiota. This study aimed to identify the salivary microbial structure in liver disease patients and determine whether the disease progression influence the bacterial composition. 16S rDNA high-throughput sequencing and bioinformatic analysis were used to examine oral bacterial diversity in the different status of hepatitis patients including 6 patients with Hepatitis B (Y), 6 patients with Hepatitis B Cirrhosis (YY) and 6 patients with liver cancer (C), and 6 healthy controls (T). Phylogenetic analysis revealed that the genera of Streptococcus, Prevotella, Actinomyces, Veillonella and Neisseria are predominant genus in the saliva of Y, YY, C patients and T group. Lautropia, Abiotrophia and Veillonella were enriched in Y patients, while Treponema, Selenomonas and Oribacterium were also existed in YY patients. Haemophilus, Porphyromonas and Filifactor had high abundance in C patients. The genera of Moryella, Leptotrichia, Lactobacillus, Dialister, Serratia, Enterococcus and Actinobacillus were decreased in all patient samples compared with healthy control samples which may be used for treatment of liver disease. Diversity analyses showed decreased diversity of salivary bacterial communities was discovered in the progress of the liver disease. These findings identified the oral microbiota dysbiosis in liver disease, which may providing available information and possible diagnostic biomarkers for liver patients.

Association between sleep duration, sleep quality and hyperlipidemia in middle-aged and older Chinese: The Dongfeng-Tongji Cohort Study.

AIMS: This study aimed to evaluate the relationship between sleep duration, sleep quality and hyperlipidemia in middle-aged and older Chinese. METHODS: We included 20,712 individuals at baseline from September 2008 to June 2010, and they were followed-up until October 2013. Hyperlipidemia was defined according to the Chinese guidelines on the prevention and treatment of dyslipidemia in adults. Sleep duration was self-reported and sleep quality was evaluated with a questionnaire that was designed according to the Pittsburgh Sleep Quality Index. Logistic regression and Cox proportional hazard models were conducted to explore the associations. RESULTS: In the cross-sectional analyses, longer sleep duration (≥10 h) was significantly associated with higher prevalence of hyperlipidemia (odds ratio (OR) = 1.17, 95% confidence interval (CI) = 1.02-1.35) after adjusting for potential confounders. The ORs of hyperlipidemia were significantly elevated among participants with impaired sleep quality (OR = 1.14, 95% CI = 1.08-1.22) and poor sleep quality (OR = 1.20, 95% CI = 1.08-1.34) when compared to those with good sleep quality. In the longitudinal analyses, compared to participants with a sleep duration of 7-<8 h, those with a sleep duration of 9-<10 h (hazard ratio (HR) = 1.19, 95% CI = 1.04-1.35) and ≥10 h (HR = 1.27, 95% CI = 1.02-1.58) showed significantly higher risk of hyperlipidemia after adjusting for potential confounders. However, no statistically significant association was found between impaired or poor sleep quality and hyperlipidemia. CONCLUSIONS: Longer sleep duration was significantly associated with higher risk of hyperlipidemia. Impaired or poor sleep quality were associated with elevated prevalence of hyperlipidemia, but not with the incidence of hyperlipidemia.

Combined effects of reproductive and hormone factors and obesity on the prevalence of knee osteoarthritis and knee pain among middle-aged or older Chinese women: a cross-sectional study.

BACKGROUND: Knee osteoarthritis (KOA) is one form of degenerative arthritis that results from the breakdown of cartilage and underlying bone. The prevalence of KOA is considerably higher in women than in men; however, the reason for this difference has not been thoroughly elucidated to date. The aim of the present study was to estimate the effects of reproductive and hormone factors and obesity on KOA prevalence among Chinese women. METHODS: The cross-sectional study included 7510 women with a mean age of 62.6 ± 8.6 years. Knee pain was defined as pain or aching stiffness on most days for at least 1 month during the past 12 months or persistent pain or aching stiffness within the past week. Clinical KOA was diagnosed based on both pain complaints and a Kellgren-Lawrence grade ≥ 2 X-ray radiograph of at least one knee. RESULTS: Oral contraceptives use (OR 1.18, 1.05-1.34), ≥3 pregnancies (1.38, 1.20-1.60), and postmenopausal hormone replacement therapy (HT) (1.59, 1.23-2.06) were positively associated with knee pain, while oral contraceptives use (1.28, 1.04-1.57), and HT (1.79, 1.21-2.65) were positively associated with clinical KOA. Obesity and oral contraceptives use showed additive and multiplicative effects on knee pain. The OR for knee pain among women with a BMI ≥24 kg/m2 and oral contraceptives use was 2.00 (1.68-2.38) compared with women with a BMI < 24 kg/m2 and no oral contraceptives use. CONCLUSIONS: A high number of pregnancies, oral contraceptives use, and HT are independent risk factors for KOA, and the effects of reproductive and hormone factors on KOA may be increased by obesity.

A randomized Phase III trial of neoadjuvant recombinant human endostatin, docetaxel and epirubicin as first-line therapy for patients with breast cancer (CBCRT01).

To further assess the efficacy and safety of recombinant human endostatin (rh-endostatin), a Phase III, multicenter, prospective, randomized, controlled clinical trial was conducted. Patients to be treated with neoadjuvant docetaxel and epirubicin (DE) or DE plus rh-endostatin (DEE) were eligible for this trial. The primary endpoint was clinical/pathological response. Secondary endpoints included adverse events and quality of life (QOL). Finally, 803 patients were enrolled and randomly assigned to receive DE (n = 402) or DEE (n = 401) regimen. After three cycles of neoadjuvant therapy, "complete response" achieved in 14.2% of patients in DEE group versus 6.7% in DE group, "partial response" achieved in 76.8% versus 71.1%, while "stable disease" in 6.0% versus 18.9%, "progressive disease" in 3.0% versus 3.2% of patients. The rate of objective response in DEE and DE group was 91.0% and 77.9%, respectively (p < 0.001). In spite of a relatively higher pathological complete response achieved following the combination therapy, no significant difference was found between two arms. Adverse events were mostly of Grades 1-2. No significant difference in adverse event and QOL was found between the two arms. In conclusion, the combination of chemotherapy and rh-endostatin achieved better outcomes than chemotherapy alone, and thus can be considered as a promising therapeutic strategy for breast cancer.

Immediate breast reconstruction with omental flap for luminal breast cancer patients: Ten clinical case reports.

RATIONALE: Luminal subtype breast cancer, accounting for 70 to 80% of all breast cancers, has been reported to be associated with good prognosis. However, for the patients with large mass or worse mass position, omental flap transplantation may provide a new option for breast reconstruction. PATIENT CONCERNS: Ten patients (6 luminal B1, 2 luminal B2, 2 luminal A), were enrolled into the study, between January 23, 2015 and August 22, 2016. The mean age was 34.6 ± 6.96 (24-44) years old. Immunohistochemistry demonstrated that the tumor cells were positive for estrogen receptor and progestrone receptor. DIAGNOSES: According to the clinicopathological features, diagnosis of breast cancer patients were made. INTERVENTIONS: Breast-conserving surgery, laparoscopic greater omentum harvest and vascular anas-tomosis were carried out orderly. Postoperative operative results, cosmetic outcomes, complications, as well as blood supply were investigated for surgery evaluation. Reasonable chemotherapy and irradia-tion were adopted to patients according to the pathological condition. OUTCOMES: We successfully accomplished breast reconstruction by omental flap transplantation, ex-cept one failed case because of the necrosis of omentum and changed to fat transplantation. The volumes and symmetry of breasts were all satisfied. The blood supply was detected to be fluent. Only one case of slight hematoma and another case of one distant metastasis were observed during fol-low-up period. No arm mordities or arm movement restriction occurred after surgery. Moreover, radia-tion therapy and chemotherapy had no clear effects on the reconstructed breast. LESSONS: Immediate breast reconstruction surgery by transplanting omental flap for luminal breast cancer patients can be considered successful based on the excellent clinic outcome.

The effects and mechanisms of SLC34A2 on maintaining stem cell-like phenotypes in CD147+ breast cancer stem cells.

The cancer stem cell (CSC) hypothesis has gained significant recognition in describing tumorigenesis. Identification of the factors critical to development of breast cancer stem cells (BCSCs) may provide insight into the improvement of effective therapies against breast cancer. In this study, we aim to investigate the biological function of SLC34A2 in affecting the stem cell-like phenotypes in BCSCs and its underlying mechanisms. We demonstrated that CD147+ cells from breast cancer tissue samples and cell lines possessed BCSC-like features, including the ability of self-renewal in vitro, differentiation, and tumorigenic potential in vivo. Flow cytometry analysis showed the presence of a variable fraction of CD147+ cells in 9 of 10 tumor samples. Significantly, SLC34A2 expression in CD147+ BCSCs was enhanced compared with that in differentiated adherent progeny of CD147+ BCSCs and adherently cultured cell line cells. In breast cancer patient cohorts, SLC34A2 expression was found increased in 9 of 10 tumor samples. By using lentiviral-based approach, si-SLC34A2-transduced CD147+ BCSCs showed decreased ability of sphere formation, cell viability in vitro, and tumorigenicity in vivo, which suggested the essential role of SLC34A2 in CD147+ BCSCs. Furthermore, PI3K/AKT pathway and SOX2 were found necessary to maintain the stemness of CD147+ BCSCs by using LY294002 or lentiviral-si-SOX2. Finally, we indicated that SLC34A2 could regulate SOX2 to maintain the stem cell-like features in CD147+ BCSCs through PI3K/AKT pathway. Therefore, our report identifies a novel role of SLC34A2 in BCSCs' state regulation and establishes a rationale for targeting the SLC34A2/PI3K/AKT/SOX2 signaling pathway for breast cancer therapy.

Comparison of analytic performances of Cellsearch and iFISH approach in detecting circulating tumor cells.

Circulating tumor cells (CTCs) have been widely used to predict the prognosis of breast cancer patients. The aim of the present study was to compare the performances of Cellsearch and immunostaining-fluorescence in situ hybridization (iFISH) in detecting CTCs in breast cancer patients. Forty-five newly diagnosed breast cancer patients and 14 healthy donors were recruited and their CTCs were detected by both Cellsearch and iFISH. Correlation between clinicopathological features and CTCs was investigated. We found that the positive rate of CTC detected by iFISH was significantly higher than by Cellsearch system (91% vs 38%). The CTC count, detected either by iFISH or Cellsearch, was not significantly associated with clinical pictures of patients with breast cancer. Therefore, we concluded that, compared to conventional Cellsearch CTC detection, in situ karyotypic identification performed by iFISH had higher detection rate. Therefore, iFISH may be more clinically useful than Cellsearch system.

Gelatin microspheres containing calcitonin gene-related peptide or substance P repair bone defects in osteoporotic rabbits.

OBJECTIVES: To investigate the therapeutic effect of gelatin microspheres containing different concentrations of calcitonin gene-related peptide (CGRP) or substance P on repairing bone defects in a rabbit osteoporosis model. RESULTS: Gelatin microspheres containing different concentrations of CGRP or substance P promoted osteogenesis after 3 months in a rabbit osteoporotic bone defective model. From micro-computed tomography imaging results, 10 nM CGRP was optimal for increasing the trabecular number and decreasing the trabecular bone separation degree; similar effects were observed with the microspheres containing 1 µM substance P. Histological analysis showed that the gelatin microspheres containing CGRP or substance P, regardless of the concentration, effectively promoted osteogenesis, and the highest effect was achieved in the groups containing 1 µM CGRP or 1 µM substance P. CONCLUSIONS: Gelatin microspheres containing CGRP or substance P effectively promoted osteogenesis in a rabbit osteoporotic bone defect model dose-dependently, though their effects in repairing human alveolar ridge defects still need further investigation.

Case report of a Li-Fraumeni syndrome-like phenotype with a de novo mutation in CHEK2.

BACKGROUND: Cases of multiple tumors are rarely reported in China. In our study, a 57-year-old female patient had concurrent squamous cell carcinoma, mucoepidermoid carcinoma, brain cancer, bone cancer, and thyroid cancer, which has rarely been reported to date. METHODS: To determine the relationship among these multiple cancers, available DNA samples from the thyroid, lung, and skin tumors and from normal thyroid tissue were sequenced using whole exome sequencing. RESULTS: The notable discrepancies of somatic mutations among the 3 tumor tissues indicated that they arose independently, rather than metastasizing from 1 tumor. A novel deleterious germline mutation (chr22:29091846, G->A, p.H371Y) was identified in CHEK2, a Li-Fraumeni syndrome causal gene. Examining the status of this novel mutation in the patient's healthy siblings revealed its de novo origin. CONCLUSION: Our study reports the first case of Li-Fraumeni syndrome-like in Chinese patients and demonstrates the important contribution of de novo mutations in this type of rare disease.

Full-length single-cell RNA-seq applied to a viral human cancer: applications to HPV expression and splicing analysis in HeLa S3 cells.

BACKGROUND: Viral infection causes multiple forms of human cancer, and HPV infection is the primary factor in cervical carcinomas. Recent single-cell RNA-seq studies highlight the tumor heterogeneity present in most cancers, but virally induced tumors have not been studied. HeLa is a well characterized HPV+ cervical cancer cell line. RESULT: We developed a new high throughput platform to prepare single-cell RNA on a nanoliter scale based on a customized microwell chip. Using this method, we successfully amplified full-length transcripts of 669 single HeLa S3 cells and 40 of them were randomly selected to perform single-cell RNA sequencing. Based on these data, we obtained a comprehensive understanding of the heterogeneity of HeLa S3 cells in gene expression, alternative splicing and fusions. Furthermore, we identified a high diversity of HPV-18 expression and splicing at the single-cell level. By co-expression analysis we identified 283 E6, E7 co-regulated genes, including CDC25, PCNA, PLK4, BUB1B and IRF1 known to interact with HPV viral proteins. CONCLUSION: Our results reveal the heterogeneity of a virus-infected cell line. It not only provides a transcriptome characterization of HeLa S3 cells at the single cell level, but is a demonstration of the power of single cell RNA-seq analysis of virally infected cells and cancers.

Comparison of variations detection between whole-genome amplification methods used in single-cell resequencing.

BACKGROUND: Single-cell resequencing (SCRS) provides many biomedical advances in variations detection at the single-cell level, but it currently relies on whole genome amplification (WGA). Three methods are commonly used for WGA: multiple displacement amplification (MDA), degenerate-oligonucleotide-primed PCR (DOP-PCR) and multiple annealing and looping-based amplification cycles (MALBAC). However, a comprehensive comparison of variations detection performance between these WGA methods has not yet been performed. RESULTS: We systematically compared the advantages and disadvantages of different WGA methods, focusing particularly on variations detection. Low-coverage whole-genome sequencing revealed that DOP-PCR had the highest duplication ratio, but an even read distribution and the best reproducibility and accuracy for detection of copy-number variations (CNVs). However, MDA had significantly higher genome recovery sensitivity (~84 %) than DOP-PCR (~6 %) and MALBAC (~52 %) at high sequencing depth. MALBAC and MDA had comparable single-nucleotide variations detection efficiency, false-positive ratio, and allele drop-out ratio. We further demonstrated that SCRS data amplified by either MDA or MALBAC from a gastric cancer cell line could accurately detect gastric cancer CNVs with comparable sensitivity and specificity, including amplifications of 12p11.22 (KRAS) and 9p24.1 (JAK2, CD274, and PDCD1LG2). CONCLUSIONS: Our findings provide a comprehensive comparison of variations detection performance using SCRS amplified by different WGA methods. It will guide researchers to determine which WGA method is best suited to individual experimental needs at single-cell level.

Lymphatic-targeted therapy following neoadjuvant chemotherapy: a promising strategy for lymph node-positive breast cancer treatment.

Neoadjuvant chemotherapy has been increasingly used to downstage breast cancer prior to surgery recently. However, in some cases, it was observed that despite sufficient regression of primary tumors, the metastatic lymph nodes remained nonresponsive. In this study, we applied lymphatic-targeted strategy to evaluate its efficacy and safety for patients presenting refractory nodes following systemic chemotherapy. A total of 318 breast cancer patients were demonstrated with lymph node metastasis by needle biopsy and given neoadjuvant chemotherapy. Two cycles later, 72 patients were observed with responsive tumors but stable nodes, 42 of which received a subcutaneous injection of lymphatic-targeted pegylated liposomal doxorubicin during the third cycle, while the remaining 30 patients were continued with former neoadjuvant therapeutic pattern and regarded as the control. Lymphatic-targeted treatment substantially increased both clinical and pathological node response rate [62 % (26/42) vs. 13 % (4/30) and 12 % (5/42) vs. 0 (0/30), respectively], and induced a higher apoptosis level of metastatic cells (median, 41 vs. 6 %), compared with the control. Moreover, a higher disease-free survival was observed after a median follow-up of 4 years (69 vs. 56 %). Inflammatory reaction surrounding injection sites was the most common side effect. Lymphatic chemotherapy has reliable efficacy and well-tolerated toxicity for breast cancer patients presenting refractory lymph nodes following neoadjuvant chemotherapy.

Three dysregulated microRNAs in serum as novel biomarkers for gastric cancer screening.

Gastric cancer (GC) is one of the most threatening diseases. The symptoms of GC are complex and hard to detect, which also contribute to the poor prognosis of GC. Besides, the current diagnosis for GC is expensive and invasive. Thus, a fast, noninvasive biomarker is urgently needed for GC screening. MicroRNAs (miRNAs) are small noncoding RNAs, which are involved in a great variety of pathological processes, particularly carcinogenesis. MiRNAs are stable in gastric juice, plasma as well as serum, which facilitate it to be a promising biomarker for cancer. In this study, we selected three novel miRNAs, i.e., miR-233, miR-16, and miR-100, to investigate their potential diagnostic value in GC screening. A total of 50 GC patients and 47 healthy controls were involved in this study. Blood serum samples were collected; RNAs were extracted and normalized with U6 snRNA as the internal control; qRT-PCR was performed for relative expression of target miRNAs. Levels of miRNAs expression were compared by Student's t test for the comparison between two groups, and one-way ANOVA was used for multiple comparisons. The expression of miR-223, miR-16, and miR-100 was all significantly higher in GC patients than controls (all P < 0.001). All the tested miRNAs were manifested to be valuable biomarkers for GC. Relative expression of these miRNAs was significantly correlated with clinical characteristics of GC patients, such as TNM stage (P = 0.036 for miR-223; P < 0.001 for miR-100), metastatic status (P = 0.045 for miR-223; P = 0.031 for miR-16; P = 0.006 for miR-100), tumor size (P = 0.042 for miR-223; P = 0.031 for miR-16; P < 0.001 for miR-100), and differentiation grade (P = 0.036 for miR-223; P = 0.030 for miR-16; P = 0.034 for miR-100). However, in T classification, which considered both tumor size and direct extent of primary tumor, the difference in target miRNAs expression was not significant. In summary, we confirmed the diagnostic value of serum miR-223, miR-16, and miR-100 in GC. Significantly elevated expression of the three miRNAs was also observed in advanced GC patients, which suggested their availability in cancer staging.

Multiplex PCR/mass spectrometry screening of biological carcinogenic agents in human mammary tumors.

BACKGROUND: While many studies have suggested a possible link between breast cancer pathogenesis and infection by viruses, the role of viruses in breast carcinogenesis remains controversial. OBJECTIVES: We analyzed the prevalence of 30 oncogenic human papillomaviruses (HPVs), Epstein-Barr virus (EBV), Kaposi's sarcoma herpes virus (KSHV) and six polyomaviruses in breast tumor specimens. STUDY DESIGN: We analyzed breast specimens from 100 breast cancer patients (group 1) and 50 benign breast disease patients (group 2) from Shaanxi Province in China. We also screened for the viruses in blood samples from the patients and 96 female blood donor volunteers (group 3). RESULTS: EBV, Merkel cell polyomavirus (MCPyV) and HPV-18 were detected in 60, 14 and 2 breast cancer patients, respectively, and EBV and MCPyV were detected in 16 and 1 benign breast disease patients, respectively. EBV and MCPyV were more prevalent in group 1 than in group 2 (EBV: 60.0% vs. 32.0%, p = 0.0012; MCPyV: 14.0% vs. 2.0%; p = 0.02). In contrast, there was no difference in the prevalence of EBV and MCPyV in blood samples between group 1 and group 2, group 1 and group 3. EBV was detected in malignant breast tissue and its presence was confined to the malignant cells using in situ hybridization. CONCLUSIONS: We found that EBV and MCPyV were more prevalent in the tumors of women with breast cancer than in samples from women with benign breast disease. Our results support an etiologic role for EBV in breast cancer pathogenesis in Chinese patients.

Subanesthetic isoflurane reduces zymosan-induced inflammation in murine Kupffer cells by inhibiting ROS-activated p38 MAPK/NF-κB signaling.

Volatile anesthetic isoflurane (ISO) has immunomodulatory effects. The fungal component zymosan (ZY) induces inflammation through toll-like receptor 2 or dectin-1 signaling. We investigated the molecular actions of subanesthetic (0.7%) ISO against ZY-induced inflammatory activation in murine Kupffer cells (KCs), which are known as the resident macrophages within the liver. We observed that ISO reduced ZY-induced cyclooxygenase 2 upregulation and prostaglandin E2 release, as determined by western blot and radioimmunoassay, respectively. ISO also reduced the production of tumor necrosis factor-α, interleukin-1ß, IL-6, high-mobility group box-1, macrophage inflammatory protein-1α, macrophage inflammatory protein-2, and monocyte chemoattractant protein-1 as assessed by enzyme-linked immunosorbent assays. ISO blocked the ZY-induced nuclear translocation and DNA-binding activity of nuclear factor- (NF)-κB p65. Moreover, ISO attenuated ZY-induced p38 mitogen-activated protein kinase (MAPK) activation partly by scavenging reactive oxygen species (ROS); the interregulation that ROS activated p38 MAPK followed by NF-κB activation was crucial for the ZY-induced inflammatory responses in KCs. An in vivo study by peritoneal injection of ZY into BALB/C mice confirmed the anti-inflammatory properties of 0.7% ISO against ZY in KCs. These results suggest that ISO ameliorates ZY-induced inflammatory responses in murine KCs by inhibiting the interconnected ROS/p38 MAPK/NF-κB signaling pathways.

Detection and analysis of human papillomavirus (HPV) DNA in breast cancer patients by an effective method of HPV capture.

Despite an increase in the number of molecular epidemiological studies conducted in recent years to evaluate the association between human papillomavirus (HPV) and the risk of breast carcinoma, these studies remain inconclusive. Here we aim to detect HPV DNA in various tissues from patients with breast carcinoma using the method of HPV capture combined with massive paralleled sequencing (MPS). To validate the confidence of our methods, 15 cervical cancer samples were tested by PCR and the new method. Results showed that there was 100% consistence between the two methods.DNA from peripheral blood, tumor tissue, adjacent lymph nodes and adjacent normal tissue were collected from seven malignant breast cancer patients, and HPV type 16 (HPV16) was detected in 1/7, 1/7, 1/7 and 1/7 of patients respectively. Peripheral blood, tumor tissue and adjacent normal tissue were also collected from two patients with benign breast tumor, and 1/2, 2/2 and 2/2 was detected to have HPV16 DNA respectively. MPS metrics including mapping ratio, coverage, depth and SNVs were provided to characterize HPV in samples. The average coverage was 69% and 61.2% for malignant and benign samples respectively. 126 SNVs were identified in all 9 samples. The maximum number of SNVs was located in the gene of E2 and E4 among all samples. Our study not only provided an efficient method to capture HPV DNA, but detected the SNVS, coverage, SNV type and depth. The finding has provided further clue of association between HPV16 and breast cancer.