A Small RNA, SaaS, Promotes Salmonella Pathogenicity by Regulating Invasion, Intracellular Growth, and Virulence Factors.

Salmonella enterica serovar Enteritidis is a common foodborne pathogen that infects both humans and animals. The S. Enteritidis virulence regulation network remains largely incomplete, and knowledge regarding the specific virulence phenotype of small RNAs (sRNAs) is limited. Here, we investigated the role of a previously identified sRNA, Salmonella adhesive-associated sRNA (SaaS), in the virulence phenotype of S. Enteritidis by constructing mutant (ΔsaaS) and complemented (ΔsaaS/psaaS) strains. SaaS did not affect S. Enteritidis; it was activated in the simulated intestinal environment (SIE), regulating the expression of virulence target genes. We discovered that it directly binds ssaV mRNA. Caco-2 and RAW 264.7 cell assays revealed that SaaS promoted S. Enteritidis invasion and damage to epithelial cells while suppressing macrophage overgrowth and destruction. Furthermore, a BALB/c mouse model demonstrated that the deletion of SaaS significantly reduced mortality and attenuated the deterioration of pathophysiology, bacterial dissemination into systemic circulation, and systemic inflammation. Our findings indicate that SaaS is required for S. Enteritidis virulence and further highlight its biological role in bacterial pathogenesis. IMPORTANCE Salmonella is a zoonotic pathogen with high virulence worldwide, and sRNAs have recently been discovered to play important roles. We explored the biological characteristics of the sRNA SaaS and developed two cell infection models and a mouse infection model. SaaS is an SIE-responsive sRNA that regulates the expression of virulence-targeted genes. Additionally, it differentially mediates invasion and intracellular growth for survival and infection of the epithelium and macrophages. We further found that SaaS enhanced bacterial virulence by promoting lethality, colonization, and inflammatory response. These findings provide a better understanding of the critical role of sRNA in bacterial virulence.

Predictive Models for Knee Pain in Middle-Aged and Elderly Individuals Based on Machine Learning Methods.

Aim: This study used machine learning methods to develop a prediction model for knee pain in middle-aged and elderly individuals. Methods: A total of 5386 individuals above 45 years old were obtained from the National Health and Nutrition Examination Survey. Participants were randomly divided into a training set and a test set at a 7 : 3 ratio. The training set was used to create a prediction model, whereas the test set was used to validate the proposed model. We constructed multiple predictive models based on three machine learning methods: logistic regression, random forest, and Extreme Gradient Boosting. The model performance was evaluated by areas under the receiver (AUC), sensitivity, specificity, positive predictive value, and negative predictive value. Additionally, we created a simplified nomogram based on logistic regression for better clinical application. Results: About 31.4% (1690) individuals were with self-reported knee pain. The logistic regression showed that female gender (odds ratio [OR] = 1.28), pain elsewhere (OR = 4.64), and body mass index (OR = 1.05) were significantly associated with increased risk of knee pain. In the test set, the logistic regression (AUC = 0.71) showed similar but slightly higher accuracy than the random forest (AUC = 0.70), while the performance of the Extreme Gradient Boosting model was less reliable (AUC = 0.59). Based on mean decrease accuracy, the most important first five predictions were pain elsewhere, waist circumference, body mass index, age, and gender. Additionally, the most important first five predictions with the highest mean decrease Gini index were pain elsewhere, body mass index, waist circumference, triglycerides, and age. The nomogram model showed good discrimination ability with an AUC of 0.75 (0.73-0.77), a sensitivity of 0.72, specificity of 0.71, a positive predictive value of 0.45, and a negative predictive value of 0.88. Conclusion: This study proposed a convenient nomogram tool to evaluate the risk of knee pain for the middle-aged and elderly US population in primary care. All the input variables can be easily obtained in a clinical setting, and no additional radiologic assessments were required.

EP4 activation ameliorates liver ischemia/reperfusion injury via ERK1/2­GSK3ß­dependent MPTP inhibition.

Prostaglandin E receptor subtype 4 (EP4) is widely distributed in the heart, but its role in hepatic ischemia/reperfusion (I/R), particularly in mitochondrial permeability transition pore (MPTP) modulation, is yet to be elucidated. In the present study, an EP4 agonist (CAY10598) was used in a rat model to evaluate the effects of EP4 activation on liver I/R and the mechanisms underlying this. I/R insult upregulated hepatic EP4 expression during early reperfusion. In addition, subcutaneous CAY10598 injection prior to the onset of reperfusion significantly increased hepatocyte cAMP concentrations and decreased serum ALT and AST levels and necrotic and apoptotic cell percentages, after 6 h of reperfusion. Moreover, CAY10598 protected mitochondrial morphology, markedly inhibited mitochondrial permeability transition pore (MPTP) opening and decreased liver reactive oxygen species levels. This occurred via activation of the ERK1/2­GSK3ß pathway rather than the janus kinase (JAK)2­signal transducers and activators of transcription (STAT)3 pathway, and resulted in prevention of mitochondria­associated cell injury. The MPTP opener carboxyatractyloside (CATR) and the ERK1/2 inhibitor PD98059 also partially reversed the protective effects of CAY10598 on the liver and mitochondria. The current findings indicate that EP4 activation induces ERK1/2­GSK3ß signaling and subsequent MPTP inhibition to provide hepatoprotection, and these observations are informative for developing new molecular targets and preventative therapies for I/R in a clinical setting.

Morphophysiological responses of detached and adhered biofilms of Pseudomonas fluorescens to acidic electrolyzed water.

Pseudomonas spp. have emerged as the main spoilage bacteria, with many strains easily forming biofilms on food-contact surfaces and causing cross-contamination. The efficacy of disinfectants against bacteria is usually tested with planktonic cells; however, the disinfection tolerance of biofilms, especially detached biofilms, remains unknown. Here, we investigated the tolerance responses of detached and adhered biofilms of Pseudomonas fluorescens to acidic electrolyzed water (AEW) by determining tolerance responses by plate counting, comparing them using a Weibull model, and verifying changes in bacterial morphology by scanning electron microscopy. The experimental data and the responses calculated using Weibull a (scale) and b (shape) parameters agreed well (R2 values: 0.974-0.999), and we found that AEW exhibited effective antimicrobial activity against P. fluorescens, with adhered biofilms were more resistant than detached biofilms and planktonic cells. Additionally, AEW increased the bacterial membrane permeability and decreased the membrane potential, intracellular ATP concentrations, and intracellular pH while also triggering the disruption of extracellular polymeric substances. These results demonstrated that the morphophysiological responses of detached and adhered biofilms differed significantly and provided information on disinfectant-resistance strategies potentially beneficial to the development of novel disinfection approaches.

Neuron-derived FGF10 ameliorates cerebral ischemia injury via inhibiting NF-κB-dependent neuroinflammation and activating PI3K/Akt survival signaling pathway in mice.

FGF10 is a member of fibroblast growth factors (FGFs). We previously showed that FGF10 protects neuron against oxygen-glucose deprivation injury in vitro; however, the effect of FGF10 in ischemic stroke in vivo is unknown. In the present study, we showed that FGF10 was mainly expressed in neurons but not astrocytes, and detected FGF10 in mouse cerebrospinal fluid. The FGF10 levels in neurons culture medium and cell lysate were much higher than those in astrocytes. FGF10 expression in brain tissue and FGF10 level in CSF were increased in mouse middle cerebral artery occlusion (MCAO) model. Administration of FGF10 into lateral cerebroventricle not only decreased MCAO-induced brain infarct volume and neurological deficit, but also reduced the number of TUNEL-positive cells and activities of Caspases. Moreover, FGF10 treatment depressed the triggered inflammatory factors (TNF-α and IL-6) and NF-κB signaling pathway, and increased phosphorylation of PI3K/Akt signaling pathway. Blockade of PI3K/Akt signaling pathway by wortmannin and Akt1/2-kinase inhibitor, partly compromised the neuroprotection of FGF10. However, blockade of PI3K/Akt signaling pathway did not impair the anti-inflammation action of FGF10. Collectively, our results demonstrate that neuron-derived FGF10 ameliorates cerebral ischemia injury via inhibiting NF-κB-dependent neuroinflammation and activating PI3K/Akt survival signaling pathway in mice.

Effect of Huannao Yicong prescription [See Text] extract on ß-amyloid precursor protein metabolic signal transduction-related protein in brain tissue of dementia model transgenic mouse.

OBJECTIVE: To observe the effect of Huannao Yicong Prescription (, HNYC, a Chinese medical compound) extract on ß-amyloid precursor protein (APP) metabolic signal transduction related protein kinase C (PKC), tyrosine amyloid protein kinase (TrKA), and glycogen synthase kinase-3 (GSK-3) in brain tissue of transgenic mouse dementia model induced by APP. METHODS: Sixty dementia model transgenic 3-month-old mice induced by APP695V717I were randomly allocated in four groups: the model group (A), the Donepezil (0.65×10(-3) g·kg(-1)·(-1))-treated group (B), and the two HNYC-treated groups (C and D) with high dosage (2.8 g·kg(-1)·(-1)) and low dosage (1.4 g·kg(-1)·(-1)) of HNYC extract, respectively, 15 mice in each group. Besides, a normal control group was set up with 15 C57BL/6J mice with the same age and genetic background as the model mice. The drugs for treatment were administered once a day by dissolving in equal-volume distilled water through gastric infusion, continued for 6 months, to mice in group A and to normal control group equal-volume distilled water was administered instead. Spatial learning and memory capacity of mice were observed by Morris water maze; their one-time escape response memory capacity was tested by diving platform; and changes of PKC, TrkA, and GSK-3 levels in hippocampus and cortex of brain were detected by Western blotting. RESULTS: HNYC extract showed significant effects on increasing the time of model mice for swimming through the flat roof and the swimming time and path in the fourth quadrant P<0.05 or P<0.01). Diving platform test showed that the latent times in Groups B and C were longer than that in Group A significantly (P <0.05 and P<0.01). Compared with the normal control group, PKC and TrkA protein expression levels in hippocampus and cortex of model mice's brain lowered significantly (P<0.01), while GSK-3 protein expression increased significantly (P<0.01); compared with Group A (the model group), hippocampal and cortical levels of PKC protein expression in the intervened groups (B-D) as well as those of TrkA in Group C were higher (P<0.01 or P<0.05), while hippocampal levels of GSK-3 in intervened groups were lower (P<0.01). CONCLUSION: HNYC extract could obviously increase the protein expressions of PKC and TrkA and decrease the expression of GSK-3 protein in brain tissue of transgenetic mice model of dementia, and regulate APP metabolic signal transduction path, and thus to suppress the production of Aß, which is one of the dominant mechanisms for improving learning/memory capacity of dementia model animals.

Effect of early intervention with extract of Huannao Yicong Decoction (还脑益聪方) on the pathologic picture of hippocampus and neurocyte apoptosis in APP transgenic mice model of dementia.

OBJECTIVE: To observe the effect of early intervention using extract of Huannao Yicong Decoction (, HYD) on the pathological picture of hippocampus, neurocyte apoptosis, and associated regulatory genes in ß-amyloid precursor protein (APP) transgenic mice model of dementia. METHODS: Sixty APP695(V7171) transgenic mice were randomly divided into four groups of 15. The model group was treated with distilled water, the positive control group was treated with donepezil (0.65 mg/kg), and the two HYD groups were treated with high dose (2.8 g/kg) and low dose (1.4 g/kg) HYD, respectively. All testing drugs were administered through gastrogavage by dissolving in equal volume of distilled water, once a day for six successive months. In addition, a normal control group with 15 healthy C57BL/6J mice of the same age and genetic background was set up with distilled water treatment. The pathologic picture of brain tissue was observed by microscopy with HE stain; the amount of apoptosis cells in the hippocampal CA1 area was detected by TUNEL; and expressions of associated genes, Bcl-2, and Bax were determined by immunohistochemical method. RESULTS: Pathologic pictures of hippocampus showed that in the model group, cells messily arranged, neurons markedly decreased, and the surrounding tissue of some cells was loosened with edema, necrosis, and widened gap with glia cells proliferation. Compared with those in the normal group, the amount of apoptosis cells in the CA1 area was increased, Bcl-2 expression decreased, and Bax expression increased significantly, with markedly reduced Bcl-2/Bax ratio in the model group. Compared to the model group, the pathological changes were significantly milder in the HYD-treated groups, showing rather regularly arranged cells, significantly increased neurons, only few denatured necrotic cells with milder edema, less proliferation of glia cells, and obviously reduced cell apoptosis in CA1 area (P<0.05 or P<0.01). Besides, Bcl-2 expression was up-regulated and Bax expression down-regulated, and Bcl-2/Bax ratio significantly increased in the two HYD groups (P<0.05 or P<0.01). CONCLUSION: Early intervention with HYD could improve the abnormal pathologic picture of hippocampus and regulate the expressions of associated genes to suppress cell apoptosis, which might be its mechanism of action in alleviating cognitive functional disorder.

[Effects of early intervention with Huannao Yicong formula effective components on behavior and cholinergic system of ß-amyloid precursor protein transgenic mice].

OBJECTIVE: To observe the effects of early intervention with effective components from a Chinese herbal formula (Huannao Yicong formula, HNYCF) on behavior and related indicators of cholinergic system in ß-amyloid precursor protein (APP) transgenic mice. METHODS: Sixty 3-month-old APP695 V717I transgenic mice were randomly divided into model group, high-dose HNYCF group (2.80 g/(kg·d)), low-dose HNYCF group (1.40 g/(kg·d)) and donepezil group (0.65 mg/(kg·d)), with 15 mice in each group. Fifteen non-transgenic mice of the same genetic background were used as normal group. The model group and normal group were fed with equal volume of distilled water by gavage. After 6-month continuous medication, the Morris water maze and the passive avoidance test were used to detect the visual spatial learning and memory ability of each mouse. Then the mice were decapitated and their cerebral cortex and hippocampus were isolated to homogenate by sonication. Contents of acetylcholine (ACh) and acetylcholinesterase (AChE) and choline acetyltransferase (ChAT) activity in the homogenate were determined by enzyme-linked immunosorbent assay, and protein contents of cerebral cortex and hippocampus were measured by Coomassie brilliant blue method. RESULTS: Compared with the model group, high- and low-dose HNYCF and donepezil hydrochloride all improved spatial learning of APP mice in the Morris water maze. The ratio of swimming distance in the central area in the high-dose HNYCF group was longer than that in the model group (P<0.05). In the passive avoidance test, high- and low-dose HNYCF and donepezil hydrochloride improved memory function of APP mice by improving the escape latency and reducing the number of errors (P<0.05, P<0.01). High- and low-dose HNYCF and donepezil hydrochloride reduced the content of AChE, increased the activity of ChAT (P<0.01, P<0.05) and improved the content of ACh in hippocampus (P<0.05); high- and low-dose HNYCF and donepezil hydrochloride increased the content of ACh in cortex (P<0.05). Donepezil hydrochloride reduced the content of AChE in cortex (P<0.05), however, high- and low-dose HNYCF had no obvious influence (P>0.05). High- and low-dose HNYCF increased the content of ChAT in cortex (P<0.05), whereas donepezil hydrochloride had no obvious influence (P>0.05). CONCLUSION: Early intervention with HNYCF effective components can improve the learning and memory ability of APP transgenic mice. The mechanism may be related to enhancing the function of cholinergic system.

[Water quality comparison between Lake Taihu and contribute river during high water-level period].

Thirty-two main rivers around Lake Taihu were sampled during high water period in 2008 to investigate river's vital impact on Lake water quality. Different forms of nitrogen and phosphorus were analyzed for the 32 river water samples according to nutrients function to algae. The aim of this study is to provide the basic information for river rehabilitation. Based on the results, the nutrients and suspended substance (SS) concentrations of Wangyu River were the highest, its water quality was below the V grade (Chinese water quality criterion). Water quality of northern rivers was also below the V grade, organic matter and enzymatically hydrolysable phosphate (EHP) concentration were the highest. The EHP concentration was 0.240 mg/L, and accounted for 62.4% of total phosphorus (TP). Nitrite nitrogen (NO2(-) -N) concentration was exceptionally high in western rivers and the average concentration was 0.270 mg/L, it was 2-3 times higher than that of other rivers. The northern rivers' nitrogen, phosphorus and organic concentration were markedly higher than those in the north Lake region, while the western rivers' nitrogen was markedly higher than that in the west Lake region. It can also be concluded that algae and suspended sediment were able to bring more phosphorus to water by comparing the ratio of total nitrogen(TN) and TP in these rivers and Lake Taihu.

Effect of traditional and integrative regimens on quality of life and early renal impairment in elderly patients with isolated systolic hypertension.

OBJECTIVE: To observe the effect of Chinese medical regimen and integrative medical regimen on quality of life and early renal impairment in elderly patients with isolated systolic hypertension (EISH). METHODS: A multi-center, randomized, double-blinded controlled trail was adopted. A total of 270 cases of EISH were randomly divided into 3 groups: Chinese medicine group (CM), combination group and Western medicine group (WM). The course of treatment was 4 weeks. The clinical blood pressure, integral of quality of life (SF-36 scale), immunoglubin G (IgG), microalbumin (mALB), beta(2)-microglobulin (beta(2)-MG), transferrin (TRF) and N-acetyl-beta'-D-glucosa-minidase (NAG) in urine were determined before and after the treatment. RESULTS: After treatment, systolic blood pressure depressed significantly in each group (P<0.05), and the combination group was superior to CM or WM group in depressing SBP (P<0.05); in each group, integral of quality of life improved in different degree, and combination group was superior to WM group in all 8 dimensions (P<0.05). The level of mALB and beta(2)-MG in urine decreased in all groups (P<0.05), and the combination group was superior to CM group or WM group in decreasing mALB (P<0.05). CONCLUSIONS: Chinese medical regimen has affirmative effect in treating EISH patients, and could lower the systolic blood pressure, improve quality of life and early renal impairment of the patients, and integrative medical regimen has superiority on account of cooperation, and deserves further study.

Traditional Chinese versus integrative treatment in elderly patients with isolated systolic hypertension: a multicenter, randomized, double-blind controlled trial.

BACKGROUND: Isolated systolic hypertension (ISH) is a common disease in elderly people, threatening their health. Traditional Chinese medicine (TCM) treatment or integrative treatment had advantages in improving quality of life and protecting target organs, but need to be proved by large evidence-based researches. OBJECTIVE: To observe the effects of TCM treatment (Jiangya Capsule) or integrative treatment (combination of Jiangya Capsule and nimodipine) on blood pressure and vasoactive agents, and their safety in elderly ISH patients. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: A multicenter, randomized, double-blind controlled trial was adopted. A total of 270 elderly ISH patients recruited from Xiyuan Hospital, and TCM Hospital and Community Health Service Centers of Yanqing County of Beijing were randomly divided into 3 groups: TCM group (Jiangya Capsule plus nimodipine simulation, 90 cases), integrative group (Jiangya Capsule plus nimodipine, 90 cases) and Western medicine (WM) group (nimodipine plus Jiangya Capsule simulation, 90 cases). They were all treated for 4 weeks. MAIN OUTCOME MEASURES: Before and after 4-week treatment, office blood pressure, 24-hour ambulatory blood pressure, serum nitric oxide (NO), and plasma endothelin-1 (ET-1), thromboxane B2 (TXB2) and 6-keto-prostaglandin 1alpha (6-keto-PGF1alpha) were detected, and safety evaluation was conducted. RESULTS: After 4-week treatment, 5 patients in TCM group were lost to follow-up and another 5 patients were excluded, and 80 patients finished the trial; 7 patients in integrative group were lost to follow-up and another 7 patients were excluded, and 76 patients finished the trial; 2 patients in WM group were lost to follow-up and another 3 patients were excluded, and 85 patients finished the trial. After treatment, systolic blood pressure (SBP) decreased in each group (P<0.05), and integrative treatment was superior to TCM or WM treatment in decreasing SBP (P<0.05). Twenty-four hour average SBP and day average SBP decreased significantly in each group, and night average SBP decreased in integrative group, and integrative treatment was superior to TCM or WM treatment in decreasing day average SBP. Serum NO and plasma 6-keto-PGF1alpha levels were elevated and plasma ET-1 and TXB(2) levels were reduced after treatment, and integrative treatment was superior to TCM or WM treatment in reducing plasma TXB(2) level. CONCLUSION: TCM treatment or integrative treatment has affirmative effects and safety in treating elderly ISH patients, and integrative treatment has superiority in improving some indexes, and deserves further study.

Acute toxicity and effects of 1-alkyl-3-methylimidazolium bromide ionic liquids on green algae.

In the present study, the acute toxicity of 1-Alkyl-3-methylimidazolium bromide ([Cnmim]Br) on the green algal Scenedesmus obliquus and Chlorella ellipsoidea was determined. The length of alkyl side chain of these imidazolium ionic liquids were C4, C6, C8, C10 and C12. The primary production of S. obliquus was also assessed after they were exposed to 0.01, 0.05, 0.10, 0.50 and 1.00 mg/L of [C10mim]Br for 96 h. The results showed that the acute toxicity of these ionic liquids was positively correlated with the alkyl chain length. Meanwhile, the concentration of the ionic liquid strongly influenced the primary production of algae. These results indicate that [Cnmim]Br with longer alkyl length have toxic effects on the green algae, and the risk to aquatic ecosystems by ionic liquid's leaking into the water body must be evaluated in the future.

[A randomized, controlled, double-blind trial of Huannao Yicong capsule in senile patients with mild cognitive impairment].

OBJECTIVE: To observe the effect and explore the mechanism of Huannao Yicong capsule in treating senile patients with mild cognitive impairment (MCI). METHODS: The investigational drugs were packed by blind method. A randomized, double-blind and controlled trial was conducted on ninety senile patients with MCI. Other forty-five senile healthy persons were recruited to the healthy control group. The ninety senile patients were randomly divided into the Huannao Yicong capsule-treated group (45 patients administered with three Huannao Yicong capsules and two aniracetam capsule analogues) and aniracetam-treated group (45 patients treated with two aniracetam capsules and three Huannao Yicong capsule analogues). Patients in the two groups were treated three times daily for 16 weeks. Memory, traditional Chinese medicine syndrome, cerebral blood flow, free radicals and inflammatory mediators, such as superoxide dismutase (SOD), malondialdehyde (MDA), acetylcholinesterase (AchE), interleukin-1alpha (IL-1alpha) and interleukin-6 (IL-6) were determined before and after the treatment. Blood lipids, including triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), apolipoprotein A-1 (ApoA-1) and apolipoprotein B-100 (ApoB-100), were detected before and after the treatment. The safety indexes, such as routine tests of blood and urine, hepatic and renal function tests and electrocardiogram (ECG) were taken before and after the treatment. RESULTS: Index score of clinical memory scale in senile healthy people was significantly higher than that in MCI patients before treatment (P<0.01), and the content of AchE, IL-1alpha and IL-6 was obviously lower (P<0.01, P<0.05), the activity of SOD was higher (P<0.05). No significant difference was found in direction memory of clinical memory scale between the two treatment groups. Other index scores of clinical memory scale and traditional Chinese medicine syndrome in patients of Huannao Yicong capsule-treated group were significantly improved as compared with those of the aniracetam-treated group (P<0.05, P<0.01). The blood flow parameters of anterior cerebral artery, posterior cerebral artery and resistant index in patients of Huannao Yicong capsule-treated group were increased significantly (P<0.01, P<0.05). Huannao Yicong capsule could significantly increase the activity of serum SOD and decrease the content of AchE, IL-1alpha and IL-6 (P<0.01, P<0.05), better than aniracetam. Furthermore, Huannao Yicong capsule could significantly improve the blood lipid, such as the level of TG, LDL-C, HDL-C, ApoA-1 and ApoB-100 (P<0.01, P<0.05), and better than aniracetam (P<0.01, P<0.05). No significant changes were found after treatment in safety indexes, such as routine tests of blood and urine, hepatic and renal function tests and ECG. CONCLUSION: Huannao Yicong capsule has better therapeutic effect than aniracetam capsule in treating senile mild cognitive impairment.