Inhibition of TREM-1 alleviates neuroinflammation by modulating microglial polarization via SYK/p38MAPK signaling pathway after traumatic brain injury.

BACKGROUND: Traumatic brain injury (TBI), as a major public health problem, is characterized by high incidence rate, disability rate, and mortality rate. Neuroinflammation plays a crucial role in the pathogenesis of TBI. Triggering receptor expressed on myeloid cells-1 (TREM-1) is recognized as an amplifier of the inflammation in diseases of the central nervous system (CNS). However, the function of TREM-1 remains unclear post-TBI. This study aimed to investigate the function of TREM-1 in neuroinflammation induced by TBI. METHODS: Brain water content (BWC), modified neurological severity score (mNSS), and Morris Water Maze (MWM) were measured to evaluate the effect of TREM-1 inhibition on nervous system function and outcome after TBI. TREM-1 expression in vivo was evaluated by Western blotting. The cellular localization of TREM-1 in the damaged region was observed via immunofluorescence staining. We also conducted Western blotting to examine expression of SYK, p-SYK and other downstream proteins. RESULTS: We found that inhibition of TREM-1 reduced brain edema, decreased mNSS and improved neurobehavioral outcomes after TBI. It was further determined that TREM-1 was expressed on microglia and modulated subtype transition of microglia. Inhibition of TREM-1 alleviated neuroinflammation, which was associated with SYK/p38MAPK signaling pathway. CONCLUSIONS: These findings suggest that TREM-1 can be a potential clinical therapeutic target for alleviating neuroinflammation after TBI.

The effect of chronic alcohol exposure on spatial memory and BDNF-TrkB- PLCγ1 signaling in the hippocampus of male and female mice.

Alcohol is a commonly used drug worldwide, and abuse of alcohol has become a serious public health problem. Alcohol consumption over time can cause cognitive deficits and memory impairment, which is thought to be associated with changes in the hippocampus. Given previously known effects of brain-derived neurotrophic factor (BDNF) in regulating synaptic plasticity and learning and memory, we investigated the effect of chronic alcohol consumption on spatial memory impairment in both sexes and changes in BDNF signaling in the hippocampus. After 4 weeks of intermittent access to 20% alcohol, memory impairment in both male and female mice was evaluated using the Morris water maze and the expression of BDNF, TrkB, phosphorylation of PLCγ1 (p-PLCγ1) and PLCγ1 in the hippocampus was examined using Western blot. As expected, females spent longer escape latencies during the training phase, and both sexes spent shorter time in the target quadrant. Furthermore, after 4 weeks 20% alcohol exposure, we found significantly decreased expression levels of BDNF in the hippocampus of female mice but increased levels in male mice. TrkB and PLCγ1 expression showed no significant change in the hippocampus of both sexes. These findings suggest that chronic alcohol exposure may induce spatial memory impairment in both sexes and opposite changes in expression of BDNF and p-PLCγ1 in the hippocampus of males and females.

Progranulin inhibits fibrosis by interacting with and up-regulating DNAJC3 during mouse skin wound healing.

Scars place a heavy burden on both individuals and society. Our previous study found that reduction of progranulin (PGRN) promotes fibrogenesis in mouse skin wound healing. However, the underlying mechanisms have not been elucidated. Here, we report that PGRN overexpression decreases the expression of profibrotic genes alpha-smooth muscle actin (αSMA), serum response factor (SRF), and connective tissue growth factor (CTGF), thereby inhibiting skin fibrosis during wound repair. Bioinformatics analysis suggested that the heat shock protein (Hsp) 40 superfamily C3 (DNAJC3) is a potential downstream molecule of PGRN. Further experiments showed that PGRN interacts with and upregulates DNAJC3. Moreover, this antifibrotic effect was rescued by DNAJC3 knockdown. In summary, our study suggests that PGRN inhibits fibrosis by interacting with and upregulating DNAJC3 during wound healing in mouse skin. Our study provides a mechanistic explanation of the effect of PGRN on fibrogenesis in skin wound healing.

MiR-574-5p promotes cell proliferation by negatively regulating small C-terminal domain phosphatase 1 in esophageal squamous cell carcinoma.

Objectives: Esophageal cancer is one of the most common cancers with high incidence and mortality rates, especially in China. MicroRNA (miRNA) can be used as a prognostic marker for various human cancers. This study aims to detect suitable miRNA markers for esophageal squamous cell carcinoma (ESCC). Materials and Methods: Our previous gene expression data of ESCC cells and the data from GSE43732 and GSE112840 were analyzed. The expression of miR-574-5p in ESCC patients and controls was analyzed by real-time quantitative PCR. The effect of miR-574-5p on proliferation was detected by real-time cell analysis (RTCA) and EdU proliferation assay after cell transfections. The target gene small C-terminal domain phosphatase 1 (CTDSP1) of miR-574-5p was validated by luciferase reporter assay and western blotting. Results: In the current study, the bioinformatics analysis found miR-574-5p up-regulated in ESCC. The qPCR assay of 26 ESCC and 13 adjacent/ normal tissues confirmed these results. We further demonstrated that miR-574-5p overexpression promoted cell proliferation. Then the dual-luciferase reporter assay and the rescue experiment suggested that CTDSP1 was a direct target of miR-574-5p. Conclusion: MiR-574-5p played an oncological role in ESCC by interacting and negatively regulating CTDSP1. These results provided a deeper understanding of the effect of miR-574-5p on ESCC.

Encapsulation and release kinetics of ethylene into "pre-formed" V-type starch and granular cold-water-soluble starch.

Ethylene gas was loaded into "pre-formed" V-type starch (V6, V7 and V8) by molecular encapsulation, and granular cold-water-soluble starch (GCWSS) was chosen as a control. The formation, structural characteristics and morphological properties of inclusion complexes (ICs) were investigated using X-ray diffraction, scanning electron microscopy, and 13C solid-state nuclear magnetic resonance spectroscopy. The V6a-type starch represented the most effective structure in encapsulating ethylene gas among different "pre-formed" V-type starches. The Avrami equation was used to describe the release kinetic properties of ethylene gas from ICs under various conditions of temperatures and relative humidities (RHs). The release of ethylene from ICs conformed to diffusion-limited release mechanisms (temperature) and first-order kinetics (RH). This could be caused by the differences in starch structure and association mechanisms between ethylene and starch single helices. Accelerated ripening experiments showed that ethylene-starch ICs could ripen bananas within 5 days, suggesting that this molecular encapsulation technology can provide precise control and targeted application of ethylene gas in the ripening of vegetables and fruits.

The complete mitochondrial genome of Sarcophila rasnitzyni (Diptera: Sarcophagidae).

Sarcophila rasnitzyni Rohdendorf and Verves, 1985 (Diptera: Sarcophagidae) is of potential significance in medicine and epidemiology. In this study, we present the mitochondrial genome of S. rasnitzyni. The full length of the mitochondrial genome is 15,321 bp (GenBank accession no. MW592359), and 13 protein-coding genes (PCGs), two ribosomal RNAs (rRNAs), 22 transfer RNAs (tRNAs), and a non-coding control region were identified. Nucleotide composition is A 38.0%, G 9.9%, C 14.9%, T 37.2%, respectively. It reveals a strong A + T bias (75.2%). Phylogenetic analysis indicates that the species-level relationship between S. rasnitzyni and S. mongolica closely clusters together, and separates clearly from the rest of species. This study provides important genetic data for further enriching our understanding of phylogenetic relationship of sarcophagids species.

Oxygen- and bubble-generating polymersomes for tumor-targeted and enhanced photothermal-photodynamic combination therapy.

As a common feature of the tumor microenvironment (TME), hypoxia significantly impedes the effects of photodynamic therapy. Moreover, for tumor combination therapy, smart responsive and well-designed nanocarriers are highlighted to co-deliver different therapeutics, enhance drug delivery into target sites, and realize stimuli-responsive drug release. Herein, oxygen- and bubble-generating polymersomes (FIMPs) were developed for tumor-targeted and enhanced photothermal-photodynamic combination therapy. FIMPs efficiently co-encapsulated manganese dioxide (MnO2) and the hydrophobic photosensitizer indocyanine green (ICG) within the hydrophobic membrane as well as the bubble-generating reagent NH4HCO3 in the internal cavity of the vesicles, and achieved pH/temperature/reduction multiple responsiveness. The CO2 bubbles generated from the decomposition of NH4HCO3via laser irradiation or acidic environment and the cleavage of the copolymer disulfide bond in the reducing TME would destroy the vesicle structure for triggering drug release. In addition, oxygen can be produced to overcome tumor hypoxia through the high reaction activity of MnO2 with endogenous H2O2. In vitro studies have shown that FIMPs achieved good photothermal conversion efficiency, promoted the generation of oxygen and reactive oxygen species (ROS), and thus effectively killed tumor cells. In vivo studies indicated that FIMPs effectively overcome the hypoxic microenvironment within tumors and significantly inhibit tumor growth with good biocompatibility. The rationally designed oxygen- and bubble-generating polymersomes have great potential to overcome the tumor hypoxia limitations for enhancing the photothermal-photodynamic combination therapeutic effect.

An enzyme-responsive Gp1a-hydrogel for skin wound healing.

Faster recovery and fewer scars are ideal wound healing. We have demonstrated that the cannabinoid receptor 2 (CB2) agonist Gp1a is beneficial to skin wound healing, which inhibits inflammation and fibrogenesis while promoting re-epithelialization. However, the systemic administration is imprecise and overqualified for a local skin wound. Herein, we prepared Gp1a-gel using triglycerol monostearate (Tm) hydrogel and detected whether the Gp1a-gel worked effectively on mouse skin excision wounds. The results showed that Gp1a-gel might sustainably increase the CB2 for at least 8 days. It decreased inflammation and fibrogenesis while promoting wound enclosure and re-epithelialization. These results suggested Gp1a-gel may utilize as a potential formulation strategy to treat the skin wound.

Protective role of histone deacetylase 4 from ultraviolet radiation-induced DNA lesions.

Ultraviolet B (UVB) exposure is a core factor that leads to skin disease or carcinogenesis through the insufficient repair of DNA lesions. UVB-induced DNA lesions are mainly removed by the nucleotide excision repair (NER) mechanism. The expression of histone deacetylase 4 (HDAC4) is altered in the skin upon UVB exposure, indicating its possible implication in UVB-induced DNA lesions repair. Here, we investigated the role of HDAC4 in the NER removal of the main classes of UVB-induced DNA lesions consisting of cyclobutane pyrimidine dimers and pyrimidine (6-4) pyrimidone photoproducts (6-4PPs). We found that UVB irradiation increased HDAC4 expression at both the mRNA and protein levels. HDAC4 interacted with NER factor XPC, which played an important role in effectively removing the UVB-induced DNA lesions. This study provides an understanding of the HDAC4 function in DNA repair, which will allow the development of efficient strategies to protect the skin from UVR-induced diseases.

Estimation of the age of human semen stains by attenuated total reflection Fourier transform infrared spectroscopy: a preliminary study.

Semen stain is one of the most important biological evidence at sexual crime scenes. Age estimation of human semen stains plays an important role in forensic work, and it is rarely studied due to lack of well-established methods. In this study, the technique called attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) coupled with advanced chemometric methods was employed to determine the age of semen stains on three different substrates: glass slides, tissues and fabric made of regenerated cellulose fibres up to 6 d. Partial least squares regression (PLSR) was used in conjunction with spectral analysis for age estimation, and the results generated high R 2 values (cross-validation: 0.81, external validation: 0.74) but a narrow margin of error for root mean square error (RMSE) (RMSE of cross-validation: 0.77 d, RMSE of prediction: 1.02 d). Additionally, our results indicated the robustness of PLSR model was not weaken by the influence of different substrates in this study. Our results indicate that ATR-FTIR, combined with chemometric methods, shows great potential as a convenient and efficient tool for age estimation of semen stains. Moreover, the method could be applied to routine forensic investigations in the future.

Evaluating the effects of causes of death on postmortem interval estimation by ATR-FTIR spectroscopy.

Estimating postmortem interval (PMI) is one of the most challenging tasks in forensic practice due to the effects of many factors. Here, attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy combined with chemometrics was utilized to evaluate the effects of causes of death when estimating PMI and to establish a partial least square (PLS) regression model, which can precisely predict PMI under different causes of death. First, the sensitivities to causes of death (brainstem injury, mechanical asphyxia, and hemorrhage shock) of seven kinds of organs were evaluated based on their degrees of cohesion and separation. Then, the liver was selected as the most sensitive organ to establish a PMI estimation model to compare the predicted deviations from different causes of death. It turns out that the cause of death has no significant effect on estimating PMI. Next, a PLS regression model was built with kidney tissues, which have the lowest sensitivity, and this model showed a satisfactory predictive ability and wide applicability. This study demonstrates the feasibility of using ATR-FTIR spectroscopy in conjunction with chemometrics as a powerful alternative for detecting changes in biochemistry and estimating PMI. A new perspective was also provided for evaluating the effect of causes of death when predicting PMI.

An investigation on annular cartilage samples for post-mortem interval estimation using Fourier transform infrared spectroscopy.

Many attempts have been made to estimate the post-mortem interval (PMI) using bioanalytical methods based on multiple biological samples. Cartilage tissues could be used as an alternative for this purpose because their rate of degradation is slower than that of other soft tissue or biofluid samples. In this study, we applied Fourier transform infrared (FTIR) spectroscopy to acquire bioinformation from human annular cartilages within 30 days post-mortem. Principal component analysis (PCA) showed that sex and causes of death have almost no impact on the overall spectral variations caused by post-mortem changes. With pre-processing approaches, several predicted models were established using a conventional machine learning method, known as the partial least square (PLS) regression. The best model achieved a satisfactory prediction with a low error of 1.49 days using the second derivative transform of 3-point smoothing and extended multiplicative scatter correction (EMSC), and the spectral regions from proteins and carbohydrates contributed greatly to the PMI prediction. This study demonstrates the feasibility of cartilage-based FTIR analysis for PMI estimation. Further work will introduce advanced algorithms for more accurate and precise PMI prediction.

Reduction of PGRN increased fibrosis during skin wound healing in mice.

Progranulin (PGRN) is a multi-functional growth factor known to be involved in regulating of development, cell cycle progression, cell motility, tumorigenesis and angiogenesis. Research has revealed that PGRN is a crucial mediator of skin wound healing. Nonetheless, the role of PGRN in the fibrosis process of cutaneous wound healing has not been identified. In the present study, mice with excisional wounds were treated with si-m-PGRN or physiological saline. We observed the expression of PGRN in intact and post-injury skin by immunohistochemistry. Tissue sections of skin around the wound were performed by hematoxylin & eosin and masson's trichrome staining. After PGRN knockdown by siRNA, the expression of PGRN, collagen I (Col I), small mothers against decapentaplegic homolog 3 (Smad3), phosphorylated Smad3 (P-Smad3), transforming growth factor (TGF)-ß1 and TGF-ß receptor I (TßRI) were detected by real-time reverse transcription polymerase chain reaction (RT-qPCR) or Western blot. PGRN mRNA and protein expressions were increased after insult and remained above that of intact skin through day 20. Down-regulation of PGRN augmented fibrosis area, skin thickness and the expression of Col I. In addition, reduction of PGRN considerably increased the expression of TGF-ß1, TßRI, Smad3 and P-Smad3. These results indicate that PGRN knockdown enhances the fibrosis degree, probably via the TGF-ß/Smad signaling pathway.

MACC1 and HGF are associated with survival in patients with gastric cancer.

Metastasis-associsated in colon cancer 1 (MACC1), a newly identified oncogene, promotes tumor cell proliferation and invasion. In the present study, the expression of MACC1, hepatocyte growth factor (HGF) and its receptor, MET proto-oncogene (c-Met), was investigated in human gastric cancer tissues and adjacent normal tissues by immunohistochemistry. The association between the expression levels of the proteins and the clinicopathological parameters of the tumors were statistically analyzed. Furthermore, lentiviral particles expressing MACC1 were used to infect the hepatic satellite cell (HSC) line LX2. The expression of α-smooth muscle actin (SMA), HGF, matrix metallopeptidase (MMP)-2 and MMP-9 in human HSCs was examined by western blotting and reverse transcription-quantitative polymerase chain reaction. Transwell assays were used to measure the effect of MACC1-infected or non-infected HSCs on the migration and invasion abilities of MKN45 and MKN74 gastric carcinoma cells in vitro. The results demonstrated that positive protein expression of MACC1, HGF and c-Met was significantly higher in human gastric cancer tissues compared with adjacent normal tissues. Positive expression of MACC1 and c-Met in gastric cancer tissues had no correlation with the sex, age, tumor location and peritoneal metastasis of patients, but was significantly correlated with tumor size, depth of tumor invasion, lymph node metastasis, TNM stage, histological differentiation, and overall (5 years) and disease-free survival (5 years). Positive expression of each MACC1, HGF and c-Met protein was demonstrated to be positively correlated with each other in human gastric cancer tissues. Western blotting results confirmed that MACC1 protein was overexpressed in MACC1-overexpressing lentivirus-infected HSCs. Overexpression of MACC1 significantly increased HGF, MMP-2, MMP-9 and α-SMA expression levels in HSCs. Results from the Transwell assays indicated an increase in the number of MKN45 or MKN74 cells migrating towards MACC1-overexpressing HSCs, compared with control HSCs. These findings suggested that MACC1 may regulate the expression of HGF, MMP-2 and MMP-9 in HSCs, and may thus promote migration and invasion of gastric carcinoma cells. MACC1, HGF and c-Met might cooperatively participate in the malignant progression of gastric cancer. In conclusion, MACC1 might serve as a useful molecular target for the diagnosis, treatment and prognosis of gastric cancer.

Association between IL-1ß polymorphisms and gastritis risk: A meta-analysis.

BACKGROUND: Helicobacter pylori (H. pylori) infection of the human stomach regularly leads to chronic gastric inflammation. The cytokine gene interleukin (IL)-1ß has been implicated in influencing the pathology of inflammation induced by H. pylori infection. Currently, several studies have been carried out to investigate the association of IL-1ß-511 (rs16944) and IL-1ß-31 (rs1143627) polymorphisms with gastritis risk; however, the results are inconsistent and inconclusive. To assess the effect of IL-1ß polymorphisms on gastritis susceptibility, we conducted a meta-analysis. METHODS: Up to March 15, 2016, 2205 cases and 2289 controls were collected from 12 published case-control studies. Summarized odds ratios and corresponding 95% confidence intervals (CIs) for IL-1ß-511 and IL-1ß-31 polymorphisms and gastritis risk were estimated using fixed- or random-effects models when appropriate. Heterogeneity was assessed by chi-squared-based Q-statistic test, and the sources of heterogeneity were explored by subgroup analyses and logistic meta-regression analyses. Publication bias was evaluated by Begg funnel plot and Egger test. Sensitivity analyses were also performed. RESULTS: The results provided evidences that the single nucleotide polymorphisms (SNPs) in IL-1ß-31 might be associated with the gastritis risk, especially in the Caucasian population, while SNPs in the IL-1ß-511 might not be. CONCLUSION: Our studies may be helpful in supplementing the disease monitoring of gastritis in the future, and additional studies to determine the exact molecular mechanisms might inspire interventions to protect the susceptible subgroups.

Gene expression profile of human esophageal squamous carcinoma cell line TE-1.

Esophageal squamous cell carcinoma (ESCC) is one of the most common and deadly causes of cancer worldwide. However, to date, the mechanisms underlying its pathogenesis remain unclear. The present study investigated the gene expression profile of human esophageal cancer cell line TE-1, a cell model for ESCC, to gain insight to the genetic regulation of this disease. Human esophageal cancer TE-1 cells and normal esophageal HET-1A cells were cultured for isolation of total RNA. Differential expression of RNA transcripts was assessed using the Agilent 4×44 K microarray, combined with real-time PCR (qRT-PCR) for validation. Classification and function of the differential genes were illustrated by bioinformatics processing including hierarchical clustering and gene ontology (GO) analysis. We identified 4,986 transcripts with differential expression (fold-change ≥1.5, P<0.05), including 2,368 up-regulated and 2,618 down-regulated transcripts. GO analysis showed that the dysregulated transcripts were associated with biological process, cellular component, and molecular function. After bioinformatic analysis of significantly regulated signaling pathways, we found these transcripts may target 35 gene pathways, including p53 signaling, glioma, ubiquitin-mediated proteolysis, insulin signaling, cell cycle, inositol phosphate metabolism, mTOR signaling, and MAPK signaling. The differentially expressed transcripts were screened between the esophageal cancer cell line TE-1 and normal esophageal cell line HET-1A, as well as their target gene pathways. Further data mining is related to prevention and treatment of esophageal cancer.

[Expression of caspase-3 and HAX-1 after cerebral contusion in rat].

OBJECTIVE: To observe the expression pattern of caspase-3 and HCLS1-associated protein X-1 (HAX-1) at different time after cerebral contusion in rat, and explore the new method for estimating the injury interval. METHODS: The cerebral contusion model was established using adult SD male rats. Then the rats were randomly allocated into 8 groups: 2 h, 6 h, 12 h, 1 d, 3 d, and 7 d after cerebral contusion, sham-operation and normal control. Expression of caspase-3 and HAX-1 protein after cerebral contusion in rat was detected by Western blotting. Laser scanning confocal microscope was used to observe the number of HAX-1 positive cells and TUNEL-stained cells after cerebral contusion. RESULTS: The expression of caspase-3 increased parallelly with the time after cerebral contusion and reached the peak value on 3 d. The expression of caspase-3 decreased gradually and still maintained a high level expression on 7 d (P < 0.05). The expression of HAX-1 positive cell went up after injury, and reached the peak value at 6 h (P < 0.05), then turned down gradually after 12 h and went out of detection after 3 d. The number of TUNEL-stained cells increased obviously at 2 h and reached the peak value on 3 d. The number of TUNEL-stained apoptotic cells decreased gradually and still maintained a high level expression on 7 d (P < 0.05). CONCLUSION: The expression of caspase-3 and HAX-1 after cerebral contusion has time sequential regularity, which may provide new evidence for forensic diagnosis of cerebral contusion interval.

[Increased Egr-1 binding to promoter induced by histone hyperacetylation promotes gdnf gene transcription].

OBJECTIVE: To investigate the mechanism of high transcription of the glial cell-line derived neurotrophic factor (gdnf) gene induced by hyperacetylation of histone H3 lysine 9 (H3K9) at its promoter region II in rat C6 glioma cells. METHODS: The acetylation level of H3K9 at Egr-1 binding site in gdnf gene promoter region II and the binding capacity of Egr-1 to its binding site in gdnf promoter were examined by ChIP-PCR in C6 astroglioma cells and normal rat astrocytes, and its changes were investigated in C6 astroglioma cells after treatment with histone acetyltransferase inhibitor curcumin or deacetylase inhibitor trichostatin A. RESULTS: Compared normal astrocytes, C6 astroglioma cells showed significantly increased acetylation level of H3K9 at Egr-1 binding site in gdnf gene promoter region II and Egr-1 binding capacity (P<0.01). Curcumin treatment significantly reduced H3K9 acetylation level at Egr-1 binding site and decreased both the binding of Egr-1 to promoter region II and gdnf mRNA levels in C6 astroglioma cells (P<0.05). Conversely, increased H3K9 acetylation at the Egr-1 binding site induced by trichostatin A significantly increased the binding of Egr-1 to promoter region II and gdnf mRNA expression levels (P<0.05). CONCLUSION: H3K9 hyperacetylation induces increased Egr-1 binding to gdnf gene promoter II, which might be the reason for the high transcription level of gdnf gene in rat C6 glioma cells.

[Time-order expression of caspase-3 and iNOS in contused human brain tissue].

OBJECTIVE: To investigate the expression of caspase-3 and iNOS in different intervals and to provide evidence for estimation of injury intervals after brain contusion in human. METHODS: Thirty cases died of serious brain injury were included into the injury groups and 5 cases died of non-brain injury were served as control group. To analyze the changes of caspase-3 and iNOS expression in brain samples at different intervals (2h, 4-8h, 10-14h, 1-2d, 3-5d, 8-11d) by immunohistochemistry and auto-image analysis system. RESULTS: The level of caspase-3 expression started to increase in 2 hours after brain contusion compared to the control group (P<0.05). The level of caspase-3 expression continued to increase in 1-2 days and maintained high level in 3-5 days compared to the control group (P<0.05), then decreased gradually. There was no statistically significant difference between the expression level of iNOS in 2 hours with the control group (P>0.05). But the expression level of iNOS began to increase in 4-8 hours after brain contusion and reached its maximum in 1-2 days, then decreased. Weak expression of iNOS still could be detected in 8-11 days. CONCLUSION: The expression of caspase-3 and iNOS can be used as effective evidence for human brain contusion interval.