Fate of florfenicol and linezolid resistance genes and their bacterial hosts during two waste treatment models in swine feedlots.

Florfenicol resistance genes (FRGs) are widely present in livestock farms. The aim of this study was to evaluate the removal efficiencies of FRGs as well as the relationships between FRGs, mobile genetic elements (MGEs) and bacterial communities during the natural drying (ND) and anaerobic digestion (AD) processes of manure treatment in swine farms by combining bacterial isolation, quantitative PCR and metagenomic approaches. Solid manure showed a higher abundance of FRGs than fresh manure and was the main contamination source of fexA and fexB in ND farms, whilst biogas slurry displayed a lower abundance of FRGs than the wastewater in AD farms. Moreover, fresh manure and wastewater showed a high abundance of optrA, and wastewater was the main contamination source of cfr in both ND and AD farms. Both optrA/fexA-positive enterococci and cfr/fexA-positive staphylococci were mainly isolated along the farms' treatment processes. The cfr-positive staphylococci were highly prevalent in wastewater (57.14 % - 100 %) and may be associated with nasal-derived cfr-positive porcine staphylococci. An increased abundance of Enterococcus, Jeotgalibaca and Vagococcus in the bacterial community structures may account for the high optrA abundance in wastewater and Jeotgalibaca may be another potential host of optrA. Furthermore, the abundance of FRG-related MGEs increased by 22.63 % after the ND process and decreased by 66.96 % in AD farms. A significant correlation was observed between cfr and ISEnfa4, whereas no significance was found between optrA and IS1216E, although IS1216E is the predominant insertion sequence involved in the transfer of optrA. In conclusion, manure and wastewater represented independent pollution sources of FRGs in swine farms. Associated MGEs might play a key role in the transfer and persistence of FRGs. The AD process was more efficient in the removal of FRGs than the ND method, nevertheless a longer storage of slurry may be required for a complete removal.

A long waiting time from diagnosis to treatment decreases the survival of non-small cell lung cancer patients with stage IA1: A retrospective study.

Objective: The prognostic effect of delayed treatment on stage IA1 non-small cell lung cancer (NSCLC) patients is still unclear. This study aimed to explore the association between the waiting time before treatment and the prognosis in stage IA1 NSCLC patients. Methods: Eligible patients diagnosed with pathological stage IA1 NSCLC were included in this study. The clinical endpoints were overall survival (OS) and cancer-specific survival (CSS). The Kaplan-Meier method, the Log-rank test, univariable, and multivariable Cox regression analyses were used in this study. Propensity score matching was used to reduce the bias of data distribution. Results: There were eligible 957 patients in the study. The length of waiting time before treatment stratified the survival in patients [<3 months vs. ≥3-months, unadjusted hazard ratio (HR) = 0.481, P = 0.007; <2 months vs. ≥2-months, unadjusted HR = 0.564, P = 0.006; <1 month vs. ≥1-month, unadjusted HR = 0.537, P = 0.001]. The 5-year CSS rates were 95.0% and 77.0% in patients of waiting time within 3 months and over 3 months, respectively. After adjusting for other confounders, the waiting time was identified as an independent prognostic factor. Conclusions: A long waiting time before treatment may decrease the survival of stage IA1 NSCLC patients. We propose that the waiting time for those patients preferably is less than one month and should not exceed two months.

[Mechanism of DATS Inhibiting Side Population Cell Proliferation in Multiple Myeloma].

OBJECTIVE: To study the effect of diallyl thiosulfinate (DATS) on the proliferation of side population (SP) cells in multiple myeloma (MM) and its mechanism. METHODS: RPMI-8226 and NCI-H929 cells were cultured, and the level of SP cells was detected by Hoechst33342 staining. The SP cells were cultured and treated with 10 µg/ml DATS, the CCK8 assay was carried out to examine the effect of DATS on the proliferation ability in SP cells, and plate colony-forming test was used to examine the colony-forming ability, the flow cytometry assay was carried out to examine the cell cycle, Western blot assay was used to examine the expression of cyclin D1, cyclin E, CDK2 and CDK4. RESULTS: SP cells were detected in RPMI-8226 and NCI-H929 cells with a proportion of 3.17±0.98 and 2.65±0.61, respectively. DATS treatment could significantly inhibit the SP cells survival in a time-dependent manner, and also could significantly inhibit the colony forming. In addition, DATS treatment could significantly induce the G1/S arrest and suppress the expression of cyclin D1, cyclin E, CDK2 and CDK4. CONCLUSION: DATS can inhibit the proliferation and colony-forming of SP cells in multiple myeloma, and induce the G1/S arrest that may be carried out via suppressing the expression of cyclin D1, cyclin E, CDK2 and CDK4.

[Expression and Significance of NLRP3, ASC and AIM2 in Patients with Acute Leukemia].

OBJECTIVE: To explore the expression and significance of NLR family, pyrin domain containing 3 (NLRP3), apoptosis associated speck like protein containing a CRAD (ASC) and absent in melanoma 2 (AIM2) of patients with acute leukemia. METHODS: The petipheral blood samples of 19 patients with ALL and 41 patients with ANLL as the AL group (each 20 cases of newly diagnosed, relapsed and complete remission group) and 20 cases of non-hematologic malignancies as the control group were collected from July 2013 to July 2014 in the First Affiliated Hospital of Gannan Medical University. The expression levels of NLRP3, ASC and AIM2 in peripheral blood plasma were determined by ELISA. RESULTS: The expression levels of NLRP3, ASC and AIM2 in plasma of control and AL complete remission groups were significantly higher than those in newly diagnosed and relapsed groups, and were with statistical significance (P < 0.05), but there were no statistical signifirance between ALL and ANLL groups (P > 0.05). CONCLUSION: The expression of NLRP3, ASC and AIM2 is down-regulated in the patients with acute leukemia, which maybe play a role of anti-leukemia, and provide a laboratory evidence for diagnosis and treatment of patients with acute leukemia.

[Expression of HMGB1 and NF-κB in Patients with Acute Leukemia and Its Significance].

OBJECTIVE: To explore the expression of high mobility group box protein 1 (HMGB1) and nuclear factor-kappa B (NF-κB) in patients with acute leukemia and its significance. METHOD: 20 samples of bone marrow and peripheral blood from each acute leukemia groups (newly diagnozed, relapsed and complete remission groups) and 20 samples as control from patients with no-hematologic malignancies were collected. The expression level of HMGB1 in peripheral blood plasma was determined by ELISA; HMGB1 and NF-κB level in mononuclear cells were examined by RT-PCR. Western blot was used to determine HMGB1 and NF-κB protein levels. HMGB1 and NF-κB in bone marrow smears were determined by immnohistochemistry method (IHC). RESULTS: The expression level of HMGB1 obviously increased in patients of newly diagnosed and relapsed groups, as compared with control group there was statistical significance (P < 0.05), but there was no obvious difference in expression level of HMGB1 between complete remission group and control group (P > 0.05). The expression level of HMGB1 and NF-kB in monnuclear cells of bone marrow in newly-diagnosed group and relapsed group was significantly higher than that in control group (P < 0.05), but the expression levels of HMGB1 and NF-kB in complete remisson group did not change (P > 0.05). The results of immnohistochemistry method indicated that the possitive expression of HMGB1 and NF-kB maily was found in bone marrow smears of newly diagnosed and relapsed groups. CONCLUSION: HMGB1 is overexpressed in acute leukemia, which may be involved in the occurrence and development of acute leukemia by activating the NF-κB signaling pathway, HMGB1 may be a important index for observing therapeutic effectiveness and predicting recurrence of acute leukemia.

[Clinical study of 32 patients with adult Philadelphia chromosome-positive acute lymphoblastic leukemia].

This study was aimed to evaluate the efficacy and safety of imatinib in the treatment of patients with adult Ph chromosome-positive acute lymphoblastic leukemia (Ph(+)ALL). A total of 32 diagnosed adult Ph(+)ALL patients from July 2007 to February 2014 in our hospital were retrospectively analyzed and were divided into two groups: imatinib plus chemotherapy group and traditional chemotherapy group. The differences between two groups were analysed in disease-free survival time (DFS), overall survival time (OS) and toxicity. The G banding technigue was used to analyse the karyotype, and the flow cytometry was applyed to detect the immune markers on surface of cells. The results showed that all patients expressed B cell and hematopietic stem/progenitor cell immune markers, out of them 21 patients (65.6%) were with myeloid antigens, 27 patients with simple Ph (+) phenotype and 5 patients with additional chromosome abnormality. The DFS and OS of the imatinib group were statistically longer than those of the traditional chemotherapy group (14.3 ± 4.7 months vs 10.7 ± 3.8 months) (P < 0.05) and 22.6 ± 6.8 months vs 10.7 ± 3.8 months) (P < 0.05)). There was no significant difference in toxic effects between two groups (P > 0.05)). It is concluded that the all cases of adult Ph(+)ALL are with B cell phenotype and express hematopietic stem/progenitor cell antigen. They often accompanied by expression of myeloid antigens and additonal chromosome abnormality in genetics. The combination of imatinib with chemotherapy can prolong remission time and survival time for patients of non-hematopietic stem cell transplantation on the basis of no notably increasing the toxic effects.