Autophagy in pulmonary macrophages mediates lung inflammatory injury via c-Src tyrosine kinase pathway activation during mechanical ventilation.

OBJECTIVE: It has been clearly demonstrated that autophagy plays a critical role in mechanical ventilation-Induced lung injury (VILI). Herein, we first evaluated the mutual effects of autophagy and c-Src signaling on the lung inflammatory response to mechanical ventilation. MATERIALS AND METHODS: Mice were respectively subjected to a lower or higher lung stretch induced by mechanical ventilation with low (7 mL/kg) or high (28 mL/kg) tidal volume, before measuring the activation of autophagy and c-Src signaling through LC3 lipidation and c-Src phosphorylation, respectively. Bone marrow-derived macrophages (BMDMs) were transfected with Atg5 siRNA and administered to AM-depleted mice to generate an autophagy-deficient phenotype, and c-Src signaling was evaluated by Western blot assay to determine the impact of autophagy on c-Src activation during VILI. Afterwards, the c-Src pathway was then blocked using PP2, prior to the evaluation of polymorphonuclear neutrophils (PMN), total cell counts in BAL fluid, and lung injury scores, in order to elucidate the role of the c-Src pathway in autophagy-mediated VILI. RESULTS: Both LC3-II and p-c-Src were remarkably increased after mechanical ventilation, in a time-dependent and tidal volume-dependent manner. Moreover, c-Src phosphorylation induced by ventilation was significantly compromised in autophagy-deficient mice. On the other hand, LC3-II expression did not change due to c-Src signaling abolishment. But the inflammatory response induced by injurious ventilation was markedly attenuated by PP2 or AM-abolishment, shown by PMN and total cell counts in BAL fluid, as well as lung injury scores. CONCLUSIONS: Our results suggested that autophagy caused VILI via regulating c-Src activation, which implies that c-Src may serve as a promising therapeutic target in VILI.

Circulating microRNA array (miR-182, 200b and 205) for the early diagnosis and poor prognosis predictor of non-small cell lung cancer.

OBJECTIVE: Non-small cell lung cancer (NSCLC) is the most common cause for cancer-related mortality worldwide. Currently, early detection of NSCLC is one of the main available strategies for improving its prognosis. Due to the lack of non-invasive and convenient tools, early diagnosis of NSCLC remains poor. Recently, it has been reported that circulating microRNAs (miRNAs) can be stably detected in serum. Meanwhile, they play a powerful role as biomarkers in various tumors. Therefore, the aim of this study was to detect the expression levels of serum miR-182, 200b and 205 in NSCLC patients, and to investigate their diagnostic and prognostic values. PATIENTS AND METHODS: Real-time quantitative polymerase chain reaction (RT-qPCR) was carried out to measure the expressions of miR-182, 200b and 205 in NSCLC tissues and normal controls. Receiver-operating characteristic (ROC) curve analysis was performed to assess the potential value of serum miRNAs for NSCLC diagnosis. Meanwhile, transwell assays were performed to observe the functional effects of miRNAs on the invasion and migration of NSCLC cells. RESULTS: Compared with normal controls, serum levels of miR-182 and 205 in NSCLC patients were significantly upregulated, whereas miR-200b was remarkably downregulated. ROC analysis indicated that miRNA array (miR-182, 200b and 205) was useful biomarkers for early diagnosis of NSCLC. In addition, transwell assays demonstrated that miR-182 promoted the invasion and migration of NSCLC cells. CONCLUSIONS: Our findings revealed that serum miR-182, 200b and 205 might serve as promising biomarkers for early detection and treatment of NSCLC.

Effects of propofol or sevoflurane anesthesia on the perioperative inflammatory response, pulmonary function and cognitive function in patients receiving lung cancer resection.

OBJECTIVE: To investigate the effects of propofol and sevoflurane anesthesia on the inflammatory response, pulmonary function and cognitive function of patients undergoing lung cancer resection and their differences. PATIENTS AND METHODS: 62 patients with lung cancer who underwent pulmonary lobectomy from January 2014 to January 2016 in Jining First People's Hospital were selected and randomly divided into two groups: the propofol group (n=31) and the sevoflurane group (n=31). Patients in the propofol group were treated with intravenous injection of propofol for anesthesia maintenance, whereas those in the sevoflurane group inhaled sevoflurane for anesthesia maintenance. All patients underwent surgical resection of the lobes by the same operator. Changes in the inflammatory response and pulmonary function of patients in the perioperative period were recorded before the induced anesthesia (t1), before one-lung ventilation (t2), after sternal closure by operation (t3) and at 24 h after operation (t4), respectively; the extubation time, eye opening time and response time of two groups of patients were recorded; mini-mental state examination (MMSE) was used to evaluate the changes in cognitive function in patients and detect the concentration of S100 calcium-binding protein ß (S100ß) in serum of patients before the induced anesthesia and at 24 h after operation, respectively. RESULTS: The difference of partial pressure of alveolar-arterial oxygen (A-aDO2), respiratory index (RI) and intra-pulmonary shunt fraction (Qs/Qt) of two groups of patients at t2 and t3 were significantly higher than those at t1 (p<0.01); during t2-t3, A-aDO2, RI and Qs/Qt of patients in the propofol group were significantly lower than those of patients in the sevoflurane group (p<0.05); the levels of interleukin-6 (IL-6) and matrix metalloproteinase-9 (MMP-9) in serum of patients after the induced anesthesia in the propofol group were significantly higher than those at t1, while the level of interleukin-10 (IL-10) was lower than that at t1 (p<0.01); during t2-t4, the levels of IL-6 and MMP-9 in serum of patients in the propofol group were significantly lower than those in patients in the sevoflurane group, while the level of IL-10 was significantly higher than that in patients in the sevoflurane group (p<0.05). The postoperative extubation time, eye opening time and response time of patients in the propofol group were significantly shorter than those of patients in the sevoflurane group (p<0.05). From intraoperative period to 24 h after operation, the prevalence rate of adverse reactions in patients in the propofol group was significantly lower than that in patients in the sevoflurane group (p<0.05); MMSE scores of two groups of patients at t4 were significantly lower than those at t1, while the concentration of S100ß was significantly higher than that at t1 (p<0.01); at t4, the MMSE score of patients in the propofol group was significantly higher than that in the sevoflurane group, while the concentration of S100ß was lower than that of patients in the sevoflurane group (p<0.05). CONCLUSIONS: Compared with sevoflurane anesthesia, propofol anesthesia can significantly reduce the perioperative inflammatory response in patients receiving lung cancer resection, shorten the recovery time after operation, protect the pulmonary function of patients, improve postoperative cognitive function, and reduce the prevalence rate of intraoperative adverse reactions.

MiR-616 promotes proliferation and inhibits apoptosis in glioma cells by suppressing expression of SOX7 via the Wnt signaling pathway.

OBJECTIVE: Accumulating evidence has indicated that miR-616 exerts tumor promoter roles in several types of cancer. However, the expression pattern and roles of miR-616 in glioma progression remain unknown. This study aimed to reveal the role of miR-616 in glioma cell proliferation and its potential mechanisms. PATIENTS AND METHODS: Real-time polymerase chain reaction was used to assay the expression of miR-616 in glioma tissue samples and glioma cell lines. MTT proliferation assay and flow cytometry analysis were performed to test the apoptosis and proliferation of glioma cell after down-regulation of miR-616. The target of miR-616 was predicted by TargetScan and confirmed by luciferase reporter assay. Changes in Wnt signaling markers expression were assessed using Western blotting. RESULTS: We found that the expression of miR-616 was increased in glioma tissues and cell lines. MTT and low cytometry analysis indicated that down-regulation of miR-616 significantly inhibited proliferation and promoted apoptosis in glioma cells. Moreover, SOX7 was confirmed to be a direct target of miR-616 in glioma cells using luciferase assay and Western blotting. Finally, it was found that down-regulation of miR-616 or upregulation of SOX7 could suppress the activity of Wnt/ß-catenin signaling in glioma cells. CONCLUSIONS: Our findings indicated that miR-616 acted as a tumor promoter in glioma, and its oncogenic roles were involved in the regulation of SOX7 and Wnt/ß-catenin signaling. Moreover, knockdown of miR-616 may provide a potential therapeutic strategy for glioma.

Association between downexpression of miR-1301 and poor prognosis in patients with glioma.

OBJECTIVE: MiR-1301 has been shown to be frequently down-regulated in various tumors. However, the clinical significance of miR-1301 in human glioma is still unclear. The aim of this study was to evaluate the prognostic impact of the expressions of miR-1301 in patients with glioma. PATIENTS AND METHODS: Quantitative Real-time PCR was used to determine the expression miR-1301 in glioma tissues and pair-matched adjacent normal tissues. The relationships between miR-1301 expression and clinicopathological parameters were examined by X2 test. Kaplan-Meier curves and multivariate Cox proportional models were used to study the impact on clinical outcome. RESULTS: We observed that miR-1301 expression was significantly lower in glioma tissues compared with adjacent non-cancerous tissues (p<0.01). Also, low expressions of miR-1301 were significantly associated with high WHO grade (p<0.006), low Karnofsky performance score (KPS) (p=0.001), and large tumor size (p=0.004). Furthermore, the data of Kaplan-Meier survival analysis showed that low miR-1301 expression significantly associated with a worse overall survival (p=0.003) and disease-specific survival (p=0.001). Finally, the univariate and multivariate analysis showed that the miR-1301 expression was an independent predictor for both overall survival and disease-specific survival in glioma. CONCLUSIONS: Our findings suggested lower miR-1301 expression resulted in poorer survival in patients with glioma, which may provide important indicators for further research.

Mutation analyses and prenatal diagnosis in families of X-linked severe combined immunodeficiency caused by IL2Rγ gene novel mutation.

We investigated the feasibility of interleukin-2 receptor gamma (IL2Rγ) gene based on gene mutation analysis and pre-natal diagnosis of X-linked severe combined immunodeficiency (X-SCID). Blood samples of patients and their parents of X-SCID (family 1) and X-SCID (family 2) were collected. IL2Rγ gene sequences of the 2 families were analyzed using bi-directional direct sequencing by polymerase chain reaction. DNA sequence changes in the IL2Rγ gene exon region and shear zone were also analyzed. We also sequenced the IL2Rγ gene in 100 healthy individuals. Prenatal genetic diagnoses for a high-risk fetus in family 1 were performed by chorionic villus sampling after determining each family's genotypes. The suspect fe-male in family 1 underwent carrier detection. Two novel mutations of IL2Rγ gene were identified, including c.361-363delGAG (p.E121del) in the patient and his mother in family 1, and c.510-511insGAACT (p.W173X) heterozygous mutation in the proband's mother in family 2. These mutations were absent in the 100 controls. Prenatal diagnosis of early pregnancy in the female fetus of family 1 was performed; the fetus was heterozygous, which was confirmed at postnatal follow-up. The suspect female in family 1 showed no mutation in carrier detection. The novel p.E121del and p.W173X mutations in IL2Rγ may have been the primary causes of disease in 2 families with X-SCID. In couples with an X-SCID reproductive history, prenatal gene mutation analysis of IL2Rγ can effectively prevent the birth of children with X-SCID and carrier detection for suspected females.

Estimation of genetic parameters for growth traits in a breeding program for rainbow trout (Oncorhynchus mykiss) in China.

Genetic parameters and breeding values for growth traits were estimated in the first and, currently, the only family selective breeding program for rainbow trout (Oncorhynchus mykiss) in China. Genetic and phenotypic data were collected for growth traits from 75 full-sibling families with a 2-generation pedigree. Genetic parameters and breeding values for growth traits of rainbow trout were estimated using the derivative-free restricted maximum likelihood method. The goodness-of-fit of the models was tested using Akaike and Bayesian information criteria. Genetic parameters and breeding values were estimated using the best-fit model for each trait. The values for heritability estimating body weight and length ranged from 0.20 to 0.45 and from 0.27 to 0.60, respectively, and the heritability of condition factor was 0.34. Our results showed a moderate degree of heritability for growth traits in this breeding program and suggested that the genetic and phenotypic tendency of body length, body weight, and condition factor were similar. Therefore, the selection of phenotypic values based on pedigree information was also suitable in this research population.

Amplification of rainbow trout microsatellites in Brachymystax lenok.

Brachymystax lenok is a cold freshwater fish accustomed to inhabit relatively high concentration of dissolved oxygen and clean upper streams. Here we present 13 polymorphic microsatellite primer pairs from rainbow trout to amplify in 32 B. lenok individuals from Ussuri River of China. The number of alleles ranged from two to seven with an average of 3.9 per locus. Observed heterozygosity ranged from 0.0625 to 0.9677. One locus showed significant deviation from Hardy-Weinberg equilibrium. These 13 loci will provide a good basis for investigation of B. lenok population structure and genetic diversity in different distribution region.