SMRT sequencing revealed the diversity and characteristics of defective interfering RNAs in influenza A (H7N9) virus infection.

Influenza defective interfering (DI) particles are replication-incompetent viruses carrying large internal deletion in the genome. The loss of essential genetic information causes abortive viral replication, which can be rescued by co-infection with a helper virus that possesses an intact genome. Despite reports of DI particles present in seasonal influenza A H1N1 infections, their existence in human infections by the avian influenza A viruses, such as H7N9, has not been studied. Here we report the ubiquitous presence of DI-RNAs in nasopharyngeal aspirates of H7N9-infected patients. Single Molecule Real Time (SMRT) sequencing was first applied and long-read sequencing analysis showed that a variety of H7N9 DI-RNA species were present in the patient samples and human bronchial epithelial cells. In several abundantly expressed DI-RNA species, long overlapping sequences have been identified around at the breakpoint region and the other side of deleted region. Influenza DI-RNA is known as a defective viral RNA with single large internal deletion. Beneficial to the long-read property of SMRT sequencing, double and triple internal deletions were identified in half of the DI-RNA species. In addition, we examined the expression of DI-RNAs in mice infected with sublethal dose of H7N9 virus at different time points. Interestingly, DI-RNAs were abundantly expressed as early as day 2 post-infection. Taken together, we reveal the diversity and characteristics of DI-RNAs found in H7N9-infected patients, cells and animals. Further investigations on this overwhelming generation of DI-RNA may provide important insights into the understanding of H7N9 viral replication and pathogenesis.

Diabetes mellitus is a recurrence-independent risk factor in patients with hepatitis B virus-related hepatocellular carcinoma undergoing resection.

OBJECTIVES: The prognostic impact of diabetes mellitus and its association with virological factors on patients with hepatocellular carcinoma (HCC) are unclear. This study aimed to investigate the outcome in HCC patients undergoing resection with and without diabetes mellitus and the interaction with hepatitis B virus (HBV) and hepatitis C virus (HCV). METHODS: A total of 239 HCC patients were included. Survival and tumour recurrence were analysed according to the status of diabetes mellitus and viral hepatitis. RESULTS: Diabetic patients tended to have an overall decreased survival compared with non-diabetic patients in 32 +/- 19 months of follow-up. A total of 113 (47%) patients developed tumour recurrence during the follow-up period. No significant difference of tumour recurrence between diabetic and non-diabetic patients, or between patients seropositive and seronegative for HBV surface antigen (HBsAg), was noted. Stratified analysis showed that diabetic patients with HBV but not with HCV infection had a poor long-term outcome. In the HBV group, the one-, three- and five-year survival rates were 83%, 51% and 41% versus 90%, 78% and 73% in diabetic and non-diabetic patients, respectively, compared with 90%, 83% and 42% versus 91%, 73% and 73% in the HCV group with and without diabetes mellitus, respectively. Cox multivariate analysis disclosed that diabetic patients seropositive for HBsAg had a significantly poor survival. CONCLUSIONS: Diabetes mellitus does not affect the long-term survival in HCV-related HCC but is a recurrence-independent poor prognostic factor for HBV-related HCC.

Herb-related aneuploidy in breast fibroadenoma.

There is consensus that attitudes toward seeking complementary and alternative medicine (CAM) are different between oriental and western patients. A 16-year-old girl who presented with enlarged breast tumours also had a 3-year history of "Si-Wu-Tang" (SWT) intake. Pathology of the resected breast tumour disclosed a giant fibroadenoma with aneuploid status. A cohort study was conducted between two groups of patients with fibroadenomas: SWT intake (+) group and SWT intake (-) group. Patients were told to discontinue SWT intake for 3 months, and this was followed by breast ultrasonographic examination in both groups. The tumour sizes before and after discontinued SWT intake were 2.3±0.11 cm and 1.5±0.12 cm in the SWT (+) group, and 1.7±0.15 cm and 1.6±0.14 cm in SWT (-) group, respectively (p<0.05). It is concluded that this report provides important information for patients with breast tissue diseases and that continuous intake of medicinal herbs is recommended only under the guidance of trained CAM providers.

Diabetes mellitus is a risk factor for hepatic decompensation in patients with hepatocellular carcinoma undergoing resection: a longitudinal study.

OBJECTIVES: Patients with hepatocellular carcinoma (HCC) frequently have diabetes mellitus (DM) due to coexisting liver cirrhosis. The aim of this study was to assess the long-term impact of DM on the hepatic regenerative ability of HCC patients undergoing surgical resection. METHODS: We retrospectively studied 245 HCC patients (210 male; age, 61 +/- 13 yr) with well-preserved liver functions undergoing resection. Forty (16%) of them were diabetic and were controlled with hypoglycemic agents. The Child-Pugh scoring system was used to evaluate the postoperative liver regeneration ability. The endpoint was the occurrence of hepatic decompensation, defined as a sustained increase in the Child-Pugh score of 2 or more points or the development of tumor recurrence. RESULTS: Seventy-five patients (31%) developed hepatic decompensation during a follow-up period of 27 +/- 18 months (range, 3-75). DM (p = 0.001), large (>3 cm) tumor size (p = 0.044), and age > 65 yr (p = 0.058) were the factors associated with hepatic decompensation in univariate analysis. Multivariate Cox regression model analysis confirmed that DM (relative risk [RR] = 2.3, 95% CI = 1.4-3.7, p = 0.001) and tumor size > 3 cm (RR = 1.7, 95% CI = 1.1-2.7, p = 0.046) were independent prognostic predictors associated with the occurrence of hepatic decompensation; the respective 3- and 5-yr cumulative rates were 53% and 64% versus 27% and 50% for diabetic and nondiabetic patients, and 24% and 41% versus 38% and 60% for patients with small (< or =3 cm) versus large (>3 cm) tumors. CONCLUSION: HCC patients with DM or large tumor size are at a cumulative increased risk for postoperative hepatic decompensation.

Deterioration of hepatic functional reserve in patients with hepatocellular carcinoma after resection: incidence, risk factors, and association with intrahepatic tumor recurrence.

Hepatocellular carcinoma (HCC) is frequently associated with liver cirrhosis. Patients with HCCs undergoing surgical resection may have declining hepatic functional reserve over time. However, the incidence and risk factors of hepatic decompensation, and its relation to postoperative tumor recurrence are unknown. This study investigated 241 HCC patients (208 male; age 61 +/- 13 years) undergoing resection with a long-term follow-up. The Child-Pugh scoring system was used to evaluate the postoperative deterioration of liver reserve, defined as a sustained increment in the Child-Pugh score by 2 or more. The 1-, 3-, and 5-year cumulative probabilities of postoperative decompensation were 14%, 32%, and 56%, respectively, during a follow-up period of 27 +/- 18 months (range 3-75 months). The average increment in Child-Pugh score was 1.4 +/- 1.1 in 2.3 +/- 1.5 years, or 0.6 point per year. Altogether, 74 (31%) patients developed postoperative hepatic decompensation during the follow-up period, 43 (58%) of whom had decompensation within 2 years of resection. Large (> 3 cm) tumor size was the only independent predictor associated with hepatic decompensation (relative risk 1.7, 95% confidence interval 1.1-2.8, p = 0.041) and was a significant risk factor for intrahepatic tumor recurrence ( p = 0.018). Patients with tumor recurrence more frequently (40% of 109 patients vs. 23% of 132 patients, p = 0.005) and more rapidly (0.8 vs. 0.4 point per year) developed hepatic decompensation than those without recurrence. In conclusion, large HCCs are closely associated with hepatic decompensation in patients after resection. Tumor recurrence may predispose to the development of hepatic decompensation in these patients.