[Effect and mechanism of Yueju Pills on breast cancer induced by DMBA in rats].

This paper aims to explore the inhibitory effect of Yueju Pills on breast cancer and decipher the underlying mechanism. A total of 92 SPF-grade SD female rats were involved in this study, and 14 of them were randomly selected into control group. The remaining 78 rats were administrated with 7,12-dimethylbenzanthracene(DMBA) by gavage to establish the breast cancer model. The modeled rats were randomized into model, tamoxifen(1.9 mg·kg~(-1)·d~(-1)), and low-and high-dose(17, 34 g·kg~(-1)·d~(-1)) Yueju Pills groups. The mental state, food intake, and activities of the rats were observed daily, and the body weight was measured on alternate days. After 12 weeks of administration, the rats were sacrificed and the tumor weight was measured. The serum estrogen and progeste-rone levels were determined by enzyme-linked immunosorbent assay. The histopathological changes of the breast and tumor were observed by hematoxylin-eosin staining. Western blot was employed to measure the protein levels of glucose transporter 1(GLUT1), lactate dehydrogenase A(LDHA), phosphofructokinase muscle(PFKM), pyruvate kinase isozyme type M2(PKM2), hexokinase 2(HK2), nuclear factor-kappaB(NF-κB), and phosphorylated NF-κB. The intestinal microbiome was examined by 16S rRNA high-throughput sequencing. The results showed that compared with the model group, high and low-dose Yueju Pills showed the tumor inhibition rate of 15.8% and 64.5%, respectively, and the low dose group had stronger inhibitory effect. Compared with the control group, the model group presented elevated the levels of estrogen and progesterone in serum. The administration of Yueju Pills lowered such ele-vation, and the low-dose group showed stronger lowering effect(P<0.05). Compared with the model group, Yueju Pills reduced the glands with increased breast tissue, the degree of breast duct expansion, the number and area of acinar cavity, the secretions, and the layers of mammary epithelial cells. Furthermore, Yueju Pills down-regulated the expression of GLUT1, LDHA, PFKM, PKM2, HK2, and NF-κB(P<0.05) and altered the diversity, composition, structure, and abundance of intestinal flora. The results showed that Yueju Pills could inhibit breast cancer by regulating the secretion of estrogen and progesterone, glycolysis, inflammatory cytokines, and intestinal flora.

[Effect of Zero Valent Iron on the Horizontal Gene Transfer of Tetracycline Resistance Genes During Anaerobic Sludge Digestion Process].

The problem of bacterial resistance has become an important issue in the area of global ecological safety and human health. Waste sludge is an important reservoir and discharge source for antibiotic resistance genes (ARGs). In this study, the quantities of seven tetracycline resistance genes (TC-ARGs), including tetA, tetC, tetG, tetM, tetO, tetW, and tetX, as well as those of class 1 integron (intI1) genes, during anaerobic sludge digestion process were comprehensively quantified by quantitative PCR (qPCR). The effects of different doses of zero valent iron (Fe0) on the decrease and increase in the quantities of TC-ARGs and intI1 genes were investigated. The influence of plasmid conjugation on the horizontal gene transfer (HGT) of the target TC-ARGs was preliminarily analyzed. The correlations between the quantities of TC-ARGs and intI1 gene have been discussed. The results showed that the quantities of TC-ARGs and intI1 genes decreased in different degrees during anaerobic sludge digestion, and the abundance of tetX gene was reduced by 2.4 orders of magnitude. When Fe0 was added, no significant reduction in the quantities of TC-ARGs and intI1 genes was observed. However, as the addition of Fe0 increased, the quantities of TC-ARGs and intI1 genes increased correspondingly, as compared to those in the control group. The results obtained from the quantities of TC-ARGs carried by plasmid DNA showed that plasmid conjugation probably promoted the HGT of TC-ARGs. A positive significant correlation was found between the quantities of tetG and intI1 genes, indicating that intI1 might play an important role in the evolution of tetG during sludge anaerobic digestion process.

[Effect of Zero Valent Iron on the Decline of Tetracycline Resistance Genes and Class 1 Integrons During Thermophilic Anaerobic Digestion of Sludge].

To investigate the effects of zero valent iron (Fe0) on the decline of antibiotic resistance genes during thermophilic anaerobic digestion of sludge, the abundances of seven tetracycline resistance genes (TC-ARGs, including tetA, tetC, tetG, tetM, tetO, tetW, and tetX) and class 1 integron gene (intI1) were quantified by quantitative PCR (qPCR). Also, the concentrations of volatile fatty acids (VFAs) were determined. The correlations between the abundances of TC-ARGs and intI1 gene and the concentrations of VFAs were discussed. The results showed that appropriate dose of Fe0 such as 0.10 g·g-1 VSS could enhance the anaerobic digestion process of sludge, and the production of total VFAs and acetic acid increased significantly. The decrease in the abundances of TC-ARGs and intI1 gene was also enhanced. However, excessive Fe0 such as 1.17 g·g-1 VSS could not further improve the reduction in the abundances of TC-ARGs and intI1 gene, probably resulted from the occurrence of horizontal gene transfer. The abundances of TC-ARGs except tetO gene, as well as intI1 gene exhibited significant negative correlation with the concentration of acetic acid, indicating that acetic acid probably had an enhanced effect on the decline of TC-ARGs and intI1 gene during thermophilic anaerobic digestion of sludge.

[Pollution Characteristics, Source Analysis and Potential Ecological Risk Assessment of Heavy Metals in Soils Surrounding a Municipal Solid Waste Incineration Plant in Shanghai].

The contents of ten heavy metals (As, Cu, Cd, Cr, Pb, Ni, Zn, Ti, Mn, and Hg) were determined in the surface soils surrounding a municipal solid waste incineration plant in Shanghai using atomic spectroscopy. The spatial distribution and sources of the detected heavy metals were studied by enrichment factor and multivariate statistical and spatial interpolation analyses. In addition, their potential ecological risk was assessed. The results showed that all heavy metals, except Hg and As, were detected with mean contents ranging from 0.399 to 4220 mg·kg-1. The mean contents of Cu, Cd, Cr, Pb, Ni, Zn, and Mn were higher than their respective background values in Shanghai. In particular, the mean content of Cd was 2.9 times its background value. The results of the Pearson's correlation, principle component, enrichment factor, and spatial distribution analyses of these heavy metals indicated that Ti, Mn, and Ni primarily originated from natural sources, while Cd, Cr, Cu, Pb, and Zn originated from industrial manufacturing, combustion, and traffic emissions. The potential ecological risk assessment showed that soils surrounding the municipal solid waste incineration plant suffered from a moderate-level risk. The mean value of the potential ecological risk index of these detected heavy metals was 108.92, of which Cd contributed as high as 79.63%, deserving much attention.

[Effect of Low-concentration Ciprofloxacin on the Nitrification and Nitrifying Microorganisms of Biofilms in Biological Aerated Filter].

Effect of low-concentration ciprofloxacin (CIP) on nitrification and nitrifying microorganisms of biofilms was studied in biological aerated filters (BAF). Quantitative PCR (qPCR) was used to determine the abundance variance of four ciprofloxacin resistance genes (CIP-ARGs) during nitrification in biofilms. The correlations between the abundances of CIP-ARGs and nitrifying microorganisms were also discussed. The results showed that CIP had little influence on the ammonium oxidation process of biofilm microorganisms, whereas inhibition of the nitrite oxidation process was found. The quantitative results of ammonium-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) including Nitrobacter and Nitrospira indicated that the inhibition on the transformation of nitrite was resulted from the inhibition on Nitrobacter and Nitrospira. In addition, little influence of CIP on the relative abundance of aac and qepA in biofilms was found, but the influence on parC and oqxB was great. The abundance of Nitrotacter exhibited significant positive correlation with the abundance of parC. Similar significant correlation was also found between the abundances of Nitrospira and oqxB. It could be speculated that the genetic elements of different nitrifying microorganisms in biofilms possibly carried CIP-ARGs.

[Adsorption characteristics of typical PPCPs onto river sediments and its influencing factors].

A batch equilibrium method was used to investigate the adsorption characteristics of ciprofloxacin (CIP), tetracycline (TC), sulfamethoxazole (SMX) and triclosan (TCS) onto Huangpu River sediments. Effects of adsorption time, initial concentration, solution pH and temperature on the adsorption process were studied. The results showed that the adsorption process of these PPCPs onto sediments was a two-step process: a rapid adsorption followed by a slow balance. The equilibrium time was about 4 h. The pH value had a significant effect on the adsorption of CIP, TC and TCS, whereas the effect on SMX adsorption was negligible. The kinetic results indicated that the adsorption processes followed the pseudo-second-order model, with adsorption rate in the range of 4.89 x 10(-3)-1.96 x 10(-2) kg x (min x mg)(-1). Adsorption isotherms were well described by the Freundlich and linear equations. As temperature increased, the amount of SMX and TC adsorbed increased, whereas CIP and TCS decreased. CIP, TC and TCS had a strong tendency to adsorb onto sediments, while the adsorption of SMX was unfavorable. When the initial concentration of PPCPs was 10 mg x L(-1), the equilibrium adsorption capacities of CIP, TC, SMX and TCS reached 702.8, 733.1, 54.7 and 695.0 mg x kg(-1), respectively.

[Investigation of pollution characteristics of erythromycin resistance genes in a sewage treatment plant and the relevant selective factors].

Occurrence and distribution of twelve pharmaceutical and personal care products (PPCPs) were investigated in a sewage treatment plant in Shanghai using solid-phase extraction combined with high-performance liquid chromatography-tandem mass spectrometry (SPE-HPLC-MS/MS). Quantitative PCR (qPCR) was used to determine the distribution and removal of seven erythromycin resistance genes (ERY-ARGs). The results showed that five PPCPs including sulfamethoxazole, erythromycin, tetracycline, carbamazepine and triclosan were detected in the collected wastewater samples with concentrations in the ranges of 24.5- 38.7, 47.5-49.2, 43.1-85.4, 2.5-3.9 and 423.2-8 973.3 ng x L(-1), respectively. During the wastewater treatment process, a significant reduction of triclosan was observed, but the removal efficiencies for the other detected PPCPs were relatively low. Additionally, all target ERY-ARGs were detected in the wastewater samples ranging from 9.28 x 10(3) (ermA) to 1.83 x 10(8) (ereA) copies x L(-1) in raw influent. Though significant reductions (1.19 log-3.97 log) of ERY-ARGs were obtained, their concentrations found in the final effluent were still high. Moreover, the concentration of ERY-ARGs exhibited significant positive correlation with the concentration of erythromycin and triclosan (P < 0.05), respectively, elucidating that erythromycin played an important role in the occurrence and spread of ERY-ARGs, while triclosan may confer cross-selection for ERY-ARGs.

Pseudomonas aeruginosa isolates of distinct sub-genotypes exhibit similar potential of antimicrobial resistance by drugs exposure.

Pseudomonas aeruginosa, a wide-spread opportunistic pathogen, often complicates clinical treatments due to its resistance to a large variety of antimicrobials, especially in immune compromised patients, occasionally leading to death. However, the resistance to antimicrobials varies greatly among the P. aeruginosa isolates, which raises a question on whether some sub-lineages of P. aeruginosa might have greater potential to develop antimicrobial resistance than others. To explore this question, we divided 160 P. aeruginosa isolates collected from cities of USA and China into distinct genotypes using I-CeuI, a special endonuclease that had previously been proven to reveal phylogenetic relationships among bacteria reliably due to the highly conserved 26-bp recognition sequence. We resolved 10 genotypes by I-CeuI analysis and further divided them into 82 sub-genotypes by endonuclease cleavage with SpeI. Eight of the 10 genotypes contained both multi-drug resistant (MDR) and less resistant isolates based on comparisons of their antimicrobial resistance profiles (ARPs). When the less resistant or susceptible isolates from different genotypes were exposed to eight individual antimicrobials, they showed similar potential to become resistant with minor exceptions. This is to our knowledge the first report to examine correlations between phylogenetic sub-lineages of P. aeruginosa and their potential to become resistant to antimicrobials. This study further alerts the importance and urgency of antimicrobial abuse control.

[Analysis for post-transplant changes of peripheral lymphocyte subsets in liver transplant receptors with HBV infection].

AIM: To investigate the characters and changes of peripheral lymphocyte subsets and cytokines in liver transplant receptors with HBV infection in short phases after liver transplantation, and to provide evidences for monitoring post-transplant immune condition of liver transplant receptors. METHODS: Peripheral lymphocyte subsets and cytokine levels in pre- and post- transplant 12 h, 3 d, 10 d, 30 d, 60 d of 20 cases of patients with HBV-associated severe hepatic diseases were investigated and analyzed, and were compared respectively with those of 22 cases of healthy adults as control (HC) with flow cytometry (FCM) and ELISA. RESULTS: The patients' accounts of peripheral lymphocyte subsets before liver transplantation were lower than those of HC significantly, but the accounts decreased significantly after transplantation 12 h. Three days later, the accounts of lymphocyte subsets increased significantly. The percentages of CD3, CD4, CD8 and NK cells got to stable stage from post-transplantation 10 d, and the absolute accounts of post-transplantation 60 d were higher than those of pre-transplantation, but were still lower than those of HC; The IFN-γ and IL-10 levels of post-transplantation 12 h increased several times and decreased after 3 days. The IL-10 levels in post-transplantation 60 d were still higher than those of HC. CONCLUSION: The absolute accounts of peripheral lymphocyte subsets increased to stable levels from post-transplantation 10 d, but were still lower than those of HC; Post-transplant immune condition was to Th2 polarization.

Preparation and characterization of activated carbon produced from rice straw by (NH4)2HPO4 activation.

Effects of different pretreatment protocols in (NH(4))(2)HPO(4) activation of rice straw on porous activated carbon evolution were evaluated. The pore structure, morphology and surface chemistry of obtained activated carbons were investigated by nitrogen adsorption, scanning electron microscopy and Fourier transform infrared spectroscopy. It was found that pretreatment combining impregnation with (NH(4))(2)HPO(4) and preoxidation could significantly affect the physicochemical properties of prepared activated carbons. The apparent surface area and total pore volume as high as 1154 m(2)/g and 0.670 cm(3)/g were obtained respectively, when combined process of impregnation followed by preoxidation at 200°C and activation at 700°C was carried out. Meanwhile, the activated carbon yield and maximum methylene blue adsorption capacity up to 41.14% and 129.5 mg/g were achieved, respectively. The results exhibited that (NH(4))(2)HPO(4) could be an effective activating agent for producing activated carbons from rice straw.

mutL as a genetic switch of bacterial mutability: turned on or off through repeat copy number changes.

Bacterial adaptation to changing environments can be achieved through the acquisition of genetic novelty by accumulation of mutations and recombination of laterally transferred genes into the genome, but the mismatch repair (MMR) system strongly inhibits both these types of genetic changes. As mutation and recombination do occur in bacteria, it is of interest to understand how genetic novelty may be achieved in the presence of MMR. Previously, we observed associations of a defective MMR genotype, 6bpΔmutL, with greatly elevated bacterial mutability in Salmonella typhimurium. To validate these observations, we experimentally converted the mutL gene between the wild-type and 6bpΔmutL in S. typhimurium and inspected the bacterial mutability status. When 6bpΔmutL was converted to mutL, the originally highly mutable Salmonella strains regained genetic stability; when mutL was converted to 6bpΔmutL, the mutability was elevated 100-fold. Interestingly, mutL cells were found to grow out of 6bpΔmutL cells; the new mutL cells eventually replaced the original 6bpΔmutL population. As conversion between mutL and 6bpΔmutL may occur readily during DNA replication, it may represent a previously unrecognized mechanism to modulate bacterial mutability at the population level, allowing bacteria to respond rapidly to changing environments while minimizing the risks associated with persistent hypermutability.

Functional impact of the hyperpolarization-activated current on the excitability of myelinated A-type vagal afferent neurons in the rat.

1. The hyperpolarization-induced, cation-selective current I(h) is widely observed in peripheral sensory neurons of the vagal and dorsal root ganglia, but the peak magnitude and voltage- and time-dependent properties of this current vary widely across afferent fibre type. 2. Using patch clamp investigations of rat isolated vagal ganglion neurons (VGN) identified as myelinated A-type afferents, we established a compendium of functional correlates between changes in membrane potential and the dynamic discharge properties of these sensory neurons as a result of the controlled recruitment of I(h) using the current clamp technique. 3. Two robust responses were observed in response to hyperpolarizing step currents: (i) upon initiation of the negative step current, there was a rapid hyperpolarization of membrane potential followed by a depolarizing voltage sag (DVS) towards a plateau in membrane potential as a result of steady state recruitment of I(h); and (ii) upon termination of the negative step current, there was a rapid return to the pretest resting membrane potential that often led to spontaneous action potential discharge. These data were strongly correlated (r(2) > 0.9) with a broad compendium of dynamic discharge characteristics in these A-type VGN. 4. In response to depolarizing step currents of increasing magnitude, the discharge frequency of the A-type VGN responded with increases in the rate of sustained repetitive discharge. Upon termination of the depolarizing step current, there was a post-excitatory membrane hyperpolarization of a magnitude that was strongly correlated with action potential discharge rate (r(2) > 0.9). 5. Application of the selective hyperpolarization-activated cyclic nucleotide gated (HCN) channel blockers ZD7288 (10 micromol/L) or CsCl (1.0 mmol/L) abolished I(h) and all of the aforementioned functional correlates. In addition to reducing the excitability of the A-type VGN to step depolarizing currents. 6. Because there is increasing evidence that the HCN channel current may represent a valid target for pharmacological intervention, the quantitative relationships described in the present study could potentially help guide the molecular and/or chemical modification of HCN channel gating properties to effect a particular outcome in VGN discharge properties, ideally well beyond merely selective blockade of a particular HCN channel subtype.

Synthesis and biological evaluation of C-12' substituted vinflunine derivatives.

A series of novel C-12' substituted vinflunine derivatives have been synthesized. Several compounds in this series possess comparable in vitro cytotoxic potency against A549 cell lines.

[The changes and significance of red cell nature-immune-adhesion function in liver diseases at different stages].

OBJECTIVE: To study the changes of red cells nature immune adhesion function (RNIAF) in liver diseases at different stages, and the feasibility of RNIAF in evaluating the severity of liver diseases. METHODS: Venous blood was extracted from 682 patients with liver disease, including cirrhosis, chronic hepatitis, acute hepatitis, hepatitis gravis, and 50 healthy blood donors as controls. Suspension of red cells in self-plasma was mixed with solution of mouse ascites carcinoma cells. A tumor cell attached with 5 red cells or 2 lymphocytes/granulocytes was counted as one rosette. The adhesion rate was calculated. Serum soluble complement receptor 1 (sCR1) was measured with a newly established sandwich enzyme-linked immunosorbent assay (ELISA). The CR1 expressed on erythrocytes has assayed by cell-ELISA. RESULTS: The expression of CR1 on erythrocytes was lower among the patients with liver diseases. Compared with that in healthy controls, RNIAF decreased among the patients with various liver diseases (all P < 0.01) in the order of acute hepatitis, chronic hepatitis, cirrhosis Child's A, cirrhosis Child's B, cirrhosis Child's C, and hepatitis gravis. CHE and PTA decreased and PT increased among the patients with cirrhosis and hepatitis gravis (all P < 0.05 or P < 0.01). RNIAF was positively correlated with CHE, PT, and PTA in patents with cirrhosis. CHE, PT and PTA remained at low levels while RNIAF almost returned to its normal level in the convalescents of hepatitis gravis. CR1 expressed on erythrocytes and sCR1 in serum in various liver diseases were decreased at different degrees, however, they changed significantly later than RNIAF. Serum sCR1 significantly increased in the patients with cirrhosis, particularly in those in grades Child's A to C. CONCLUSION: Low-cost, easy to operate and with early change and stable results, RNIAF may be used as an important indicator to evaluate the severity of liver diseases. of CHE, PT/PTA. It is an important to evaluate the severity of liver function in clinic.