Frailty, Illness Perception and Lung Functional Exercise Adherence in Lung Cancer Patients After Thoracoscopic Surgery.

Background: Lung cancer patients will have lung damage after surgery, need rehabilitation exercise. Common-sense model has shown the impact of patients' perception of illness on health behaviors. However, for patients with lung cancer after thoracoscopic surgery, there has been no relevant exploration of disease perception. Objective: The purpose of this study was to investigate the clinical status of patients with lung cancer patients who have undergone thoracoscopic surgery, and to explore the correlation between frailty, disease perception, and lung functional exercise compliance. Methods: The cross-sectional study included 218 patients with lung cancer after thoracoscopic surgery. We collected participants' frailty, disease perception, exercise adherence, and relevant clinical information. T-test, Chi-square, Linear regression, Pearson's correlation, and mediation analysis were used for statistical analysis of patient data. Results: We analyzed the data by disease perception with high and low median scores and found significant differences in lymphatic dissection, stool within three days, pain, thoracic drainage tube placement time. Linear regression results show that, after controlling for confounding factors, frailty and disease perception were significantly associated with pulmonary function exercise compliance. The higher the frailty score, the worse the compliance, and the higher the disease perception negative score, the less exercise. Illness perception played a partially mediating role in the association between frailty and lung functional exercise adherence. Conclusion: Frailty and disease perception have an impact on exercise adherence, therefore, we need to consider these factors in the intervention to improve exercise compliance after thoracoscopic surgery for lung cancer.

Protocol for engineering E. coli Nissle 1917 to diagnose, record, and ameliorate inflammatory bowel disease in mice.

Engineered microorganisms hold potential for disease diagnosis and treatment. Here, we present a protocol to engineer E. coli Nissle 1917 strain (iROBOT) using genome insertion and plasmid construction to diagnose, record, and ameliorate inflammatory bowel disease in mice. We describe steps for constructing and administering iROBOT, diagnosing and recording colitis, preparing samples, and analyzing fluorescence and base editing ratios of iROBOT. We detail a colitis ameliorating assay using the disease activity index, colon length, tissue pathological section, and cytokine analysis. For complete details of the use and execution of this protocol, please refer to Zou et al.1.

Biomarker-responsive engineered probiotic diagnoses, records, and ameliorates inflammatory bowel disease in mice.

Rapid advances in synthetic biology have fueled interest in engineered microorganisms that can diagnose and treat disease. However, designing bacteria that detect dynamic disease-associated biomarkers that then drive treatment remains difficult. Here, we have developed an engineered probiotic that noninvasively monitors and records inflammatory bowel disease (IBD) occurrence and progression in real time and can release treatments via a self-tunable mechanism in response to these biomarkers. These intelligent responsive bacteria for diagnosis and therapy (i-ROBOT) consists of E. coli Nissle 1917 that responds to levels of the inflammatory marker thiosulfate by activating a base-editing system to generate a heritable genomic DNA sequence as well as producing a colorimetric signal. Fluctuations in thiosulfate also drive the tunable release of the immunomodulator AvCystatin. Orally administering i-ROBOT to mice with colitis generated molecular recording signals in processed fecal and colon samples and effectively ameliorated disease. i-ROBOT provides a promising paradigm for gastrointestinal and other metabolic disorders.

Prebiotics-Controlled Disposable Engineered Bacteria for Intestinal Diseases.

As a new method of diagnosis and treatment for intestinal diseases, intelligent engineered bacteria based on synthetic biology have been developed vigorously in recent years. However, how to deal with the engineered bacteria in vivo after completing the tasks is an urgent problem to be resolved. In this study, we constructed a thiosulfate (a biomarker of inflammatory bowel disease)-responsive engineered bacteria to generate two signals, sfGFP (monitoring) and gain-of-function (translation activation) mutation (ACG to ATG), in the initiation codon of lysisE (recording) via the CRISPR/Cas9-mediated base editing system. Once these two signals were detected, xylose could be added to induce lysis E expression, resulting in the destruction of the edited bacteria and the release of AvCystain simultaneously. Overall, our innovative engineered bacteria can record instant and historical information of the disease, and especially, the edited bacteria can be artificially attenuated and release drug in situ when needed, ultimately serving as a disposable and recyclable candidate for more types of diseases.

Exogenous salicylic acid improves resistance of aphid-susceptible wheat to the grain aphid, Sitobion avenae (F.) (Hemiptera: Aphididae).

Salicylic acid (SA), a phytohormone, has been considered to be a key regulator mediating plant defence against pathogens. It is still vague how SA activates plant defence against herbivores such as chewing and sucking pests. Here, we used an aphid-susceptible wheat variety to investigate Sitobion avenae response to SA-induced wheat plants, and the effects of exogenous SA on some defence enzymes and phenolics in the plant immune system. In SA-treated wheat seedlings, intrinsic rate of natural increase (rm), fecundity and apterous rate of S. avenae were 0.25, 31.4 nymphs/female and 64.4%, respectively, and significantly lower than that in the controls (P < 0.05). Moreover, the increased activities of phenylalanine-ammonia-lyase, polyphenol oxidase (PPO) and peroxidase in the SA-induced seedlings obviously depended on the sampling time, whereas activities of catalase and 4-coumarate:CoA ligase were suppressed significantly at 24, 48 and 72 h in comparison with the control. Dynamic levels of p-coumaric acid at 96 h, caffeic acid at 24 and 72 h and chlorogenic acid at 24, 48 and 96 h in wheat plants were significantly upregulated by exogenous SA application. Nevertheless, only caffeic acid content was positively correlated with PPO activity in SA-treated wheat seedlings (P = 0.031). These findings indicate that exogenous SA significantly enhanced the defence of aphid-susceptible wheat variety against aphids by regulating the plant immune system, and may prove a potential application of SA in aphid control.

Rosiglitazone promotes glucose metabolism of GIFT tilapia based on the PI3K/Akt signaling pathway.

The study aimed to explore the effects of rosiglitazone on glucose metabolism of GIFT tilapia based on the PI3K/Akt signaling pathway. The experiment was divided into five groups: normal starch group (32%, LC), high starch group (53%, HC), high starch +rosiglitazone group 1 (10 mg/kg, R1), high starch + rosiglitazone group 2 (20 mg/kg, R2), and high starch + rosiglitazone group 3 (30 mg/kg, R3). The results showed that a high starch diet supplemented with 10-20 mg/kg rosiglitazone had a better specific growth rate and protein efficiency that was beneficial for the growth of the tilapia. Rosiglitazone had no significant effect on the contents of crude lipid, crude protein, crude ash, and moisture of the whole fish body (p > 0.05). The contents of triglycerides and total cholesterol in the R1, R2, and R3 groups were lower than those in the HC group. The levels of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) in R1 and R2 groups were significantly lower than those in the HC groups (p < 0.05). However, the GOT and GPT levels in the R3 groups were significantly higher than those in the R1 and R2 groups (p < 0.05). With an increase in the rosiglitazone concentration, the contents of serum glucose, insulin, and hepatic glycogen in the R1, R2, and R3 groups decreased gradually. Meanwhile, the muscle glycogen content in the R1, R2, and R3 groups increased gradually. The mRNA expression of the IRS-1, PI3K, GLUT-4, and Akt proteins in the R1, R2, and R3 groups was significantly higher than that in the HC group (p < 0.05). Compared with the HC group, the expression of the GSK-3 mRNA in the R1, R2, and R3 groups was significantly reduced (p < 0.05). The protein expression of p-Akt in the R1 and R2 groups was higher than that in the HC group (p > 0.05). The protein expression of p-GSK-3ß in the R1 and R2 groups was significantly higher than that in the HC group (p < 0.05). In conclusion, a high starch diet supplemented with rosiglitazone can improve growth, enhance the serum biochemical indices, and increase the muscle glycogen content in the GIFT tilapia. It benefits in upregulating the IRS-1, PI3K, and GLUT-4 mRNA levels in the skeletal muscle and promotes glucose uptake. Meanwhile, the phosphorylation of Akt and GSK-3ß increased significantly and resulted in the inactivation of GSK-3ß and alleviation of insulin resistance.

[Effects of high glucose induced primary cardiomyocytes injury on necroptosis and the related mechanism].

OBJECTIVE: To observe whether necroptosis was happened in high glucose (HG) - induced primary cardiomyocytes injury and to investigate the likely mechanism. METHODS: The primary cultured cardiomyocytes were divided into 4 groups (n=9): control group (the cardiomyocytes were incubated with 5.5 mmol/L glucose for 48 h), HG group (the cardiomyocytes were incubated with 30 mmol/L glucose for 48 h), HG + necrostatin-1 (Nec-1) group (the cardiomyocytes was co-incubated with necroptosis inhibitor Nec-1 at 100 µmol/L and HG for 48 h) and hypertonic pressure group (HPG, the cardiomyocytes was co-incubated with 5.5 mmol/L glucose and 24.5 mmol/L mannitol for 48 h). Cell viability was measured by MTT method, reactive oxygen species (ROS) generation was measured by DHE staining. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1ß (IL-1ß) were tested by ELISA method. The mRNA and protein expressions of necroptosis related genes receptor interacting serine/threonine protein kinase 1 (RIP1), RIP3, mixed lineage kinase domain-like protein (MLKL) were tested by quantitative real-time PCR and Western blot. RESULTS: The results showed HG intervention decreased cardiomyocytes viability, increased ROS generation, up-regulated the levels of TNF-α, IL-6 and IL-1ß, increased RIP1, RIP3, MLKL expressions at mRNA and protein levels. Nec-1 treatment attenuated HG-induced increased cardiomyocytes viability, reduced ROS generation, down-regulated the levels of TNF-α, IL-6 and IL-1ß, decreased RIP1, RIP3, MLKL expressions at mRNA and protein levels. CONCLUSION: Necroptosis was happened in high glucose-induced primary cardiomyocytes injury. Inhibition of necroptosis can reduce high glucose-induced cardiomyocytes damage, may be related to inhibition of oxidative stress and depression of inflammative factors releasing.

[Effect of high concentration glucose on the expression of ALDH2 in cardiac fibroblasts of rats].

OBJECTIVE: To observe whether acetaldehyde dehydrogenase 2 (ALDH2) is expressed in cardiac fibroblasts and investigate the change of ALDH2 in cardiac fibroblasts when cultured with high concentration of glucose. METHODS: Cultured cardiac fibroblasts were randomly divided into four groups:normal control group (5.5 mmol/L glucose), Alda-1 (the agonist of ALDH2, 20µmol/L) group, high glucose group (30 mmol/L glucose) and high glucose + Alda-1 group. Cardiac fibroblasts were identified by immunofluore-scence technique. Cell prolifera-tion was detected by MTT method after treated with drugs for 48 hours. mRNA and protein expressions of ALDH2 were determined by RT-PCR and Western blot, aimed to ensure whether ALDH2 was expressed in cardiac fibroblasts. The changes of ALDH2 protein expression in cardiac fibroblasts were tested by Western blot. RESULTS: RT-PCR and Western blot results revealed that ALDH2 was expressed in cardiac fibroblasts. Compared with normal control group, cardiac fibroblasts proliferation was increased (P < 0.05), while the protein expression of ALDH2 was reduced (P < 0.05) in high glucose group. When treated with Alda-1, the proliferation of cardiac fibroblasts was decreased (P < 0.01), while the protein expression of ALDH2 was increased (P < 0.05) in high glucose group. CONCLUSIONS: ALDH2 was expressed in cardiac fi-broblasts. Alda-1, the agonist of ALDH2 enhanced the expression of ALDH2 and inhibited the proliferation of cardiac fibroblasts when cultured with high concentration of glucose.

[Role of autophagy in fasudil-induced Rho kinase inhibition for protection against myocardial ischemia-reperfusion injury in rats].

OBJECTIVE: To investigate the changes of autophagy in ischemic myocardium of rats treated with fasudil for inhibiting Rho kinase. METHODS: The hearts isolated from male Sprague-Dawley rats were subjected to 30 min of occlusion of the left anterior descending artery followed by 120 min of reperfusion with or without treatment with fasudil or fasudil+Wort. The left ventricular hemodynamics were continuously recorded, and the coronary effluent was collected during the reperfusion to determine lactate dehydrogenase (LDH) levels. The mRNA expressions of autophagy-related genes Atg5 and Beclin1 and apoptosis-related genes bax and bcl-2 were detected by RT-PCR, and the protein expression of caspase-3 was detected by Western blotting. RESULTS: Compared with I/R group, fasudil significantly improved the left ventricular developed pressure, maximal rise/fall rate of left ventricular pressure and rate pressure product, reduced LDH release during reperfusion, increased Atg5 and Beclin1 mRNA expression and the ratio of Bcl-2/Bax, and lowered caspase 3 protein expression. The autophagy inhibitor Wort significantly attenuated the effect of fasudil in the rat hearts. CONCLUSION: Fasudil treatment for inhibiting Rho kinase promoted autophagy in ex vivo rat heart to protect against myocardial ischima-reperfusion injury possibly by reducing apoptosis of the cardiac myocytes.

The Protective Effect of Low-Dose Ethanol on Myocardial Fibrosis through Downregulating the JNK Signaling Pathway in Diabetic Rats.

Objective. To investigate the effects of low dose ethanol feeding in diabetic rats and analyze its underlying mechanisms. Methods. Male Sprague-Dawley rats were divided into 4 groups: control (Con), diabetes at 4 weeks (DM4W), diabetes at 8 weeks (DM8W), and EtOH + DM8W. After 8 weeks, hemodynamic parameters were recorded and heart weight/body weight (H/B) and hydroxyproline (Hp) content in myocardium were measured. Morphology of collagen in myocardial tissue was observed with Masson's trichrome staining method and collagen volume fraction (CVF) was analysed. The mRNA expression of ALDH2 was assessed with Real-Time PCR. The protein expressions of p-JNK and JNK were evaluated using western blot. Results. In contrast to Con group, there was no difference in hemodynamic parameters in DM4W group, but mean arterial pressure and heart rate were decreased in DM8W group, and the ratios of H/B, Hp, and CVF were markedly increased. ALDH2 mRNA expression was decreased, while the ratio of p-JNK/JNK were increased. Compared with DM8W group, the above indexes were improved in EtOH + DM8W group. Conclusion. With low dose ethanol intervention, enhanced ALDH2 expression can antagonize the happening of myocardial fibrosis in diabetic rats, which may be relevant with downregulating the JNK pathway.

Long non-coding RNAs are differentially expressed in hepatocellular carcinoma cell lines with differing metastatic potential.

BACKGROUND: Metastasis is a major reason for poor prognosis in patients with cancer, including hepatocellular carcinoma (HCC). A salient feature is the ability of cancer cells to colonize different organs. Long non-coding RNAs (lncRNAs) play important roles in numerous cellular processes, including metastasis. MATERIALS AND METHODS: In this study, the lncRNA expression profiles of two HCC cell lines, one with high potential for metastasis to the lung (HCCLM3) and the other to lymph nodes (HCCLYM-H2) were assessed using the Arraystar Human LncRNA Array v2.0, which contains 33,045 lncRNAs and 30,215 mRNAs. Coding-non-coding gene co-expression (CNC) networks were constructed and gene set enrichment analysis (GSEA) was performed to identify lncRNAs with potential functions in organ-specific metastasis. Levels of two representative lncRNAs and one representative mRNA, RP5-1014O16.1, lincRNA-TSPAN8 and TSPAN8, were further detected in HCC cell lines with differing metastasis potential by qRT-PCR. RESULTS: Using microarray data, we identified 1,482 lncRNAs and 1,629 mRNAs that were differentially expressed (≥1.5 fold-change) between the two HCC cell lines. The most upregulated lncRNAs in H2 were RP11-672F9.1, RP5-1014O16.1, and RP11-501G6.1, while the most downregulated ones were lincRNA-TSPAN8, lincRNA-CALCA, C14orf132, NCRNA00173, and CR613944. The most upregulated mRNAs in H2 were C15orf48, PSG2, and PSG8, while the most downregulated ones were CALCB, CD81, CD24, TSPAN8, and SOST. Among them, lincRNA-TSPAN8 and TSPAN8 were found highly expressed in high lung metastatic potential HCC cells, while lowly expressed in no or low lung metastatic potential HCC cells. RP5- 1014O16.1 was highly expressed in high lymphatic metastatic potential HCC cell lines, while lowly expressed in no lymphatic metastatic potential HCC cell lines. CONCLUSIONS: We provide the first detailed description of lncRNA expression profiles related to organ-specific metastasis in HCC. We demonstrated that a large number of lncRNAs may play important roles in driving HCC cells to metastasize to different sites; these lncRNAs may provide novel molecular biomarkers and offer a new basis for combating metastasis in HCC cases.

Toxicity evaluation of CdTe quantum dots with different size on Escherichia coli.

Quantum dots (QDs) have a great potential for applications in nanomedicine. However, a few studies showed that they also exhibited toxicity. We used Escherichia coli (E. coli) as the model to study the effect of CdTe QDs on the cell growth by microcalorimetric technique, optical density (OD(600)) and attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectra. Three size aqueous-compatible CdTe QDs with maximum emission of 543 nm (green-emitting QDs, GQDs), 579 nm (yellow-emitting QDs, YQDs) and 647 nm (red-emitting QDs, RQDs) were tested. The growth rate constants (k) and half-inhibiting concentration (IC(50)) were calculated from the microcalorimetric data. The results indicated that CdTe QDs exhibited a dose-dependent inhibitory effect on cell growth. The order of toxicity is GQDs>YQDs>RQDs. The smaller the particle size of QDs is, the more toxicity it is. ATR-FTIR spectra indicated that the outer membrane of the cell was changed or damaged by the QDs, which may induce QDs and harmful by-products to enter into the cells. These could be one of the reasons that CdTe QDs have cytotoxic effects on E. coli.