Progress on SOX9 and its enhancers in mammalian sex determination.

The sex determination in mammals refers to the development of an initial bipotential organ, termed the bipotential gonad/genital ridge, into either a testis or an ovary at the early stages of embryonic development, under the precise regulation of transcription factors. SOX9 (SRY-box transcription factor 9) is a multifunctional transcription factor in mammalian development and plays a critical role in sex determination and subsequent male reproductive organs development. Recent studies have shown that several enhancers upstream of SOX9 also play an important role in the process of sex determination. In this review, we summarize the progress on the role of SOX9 and its gonadal enhancers in sex determination. This review will facilitate to understand the regulatory mechanism of sex determination of SOX9 and provides a theoretical basis for the further development of animal sex manipulation technologies.

Editing MC1R in human melanoma cells by CRISPR/Cas9 and functional analysis.

MC1R (melanocortin 1 receptor) encodes the melanocortin-1 receptor, which can activate intracellular cAMP synthesis under the stimulation of the α-melanocyte stimulating hormone (α-MSH) ligand. Increased cAMP then activates the protein kinase A (PKA) pathway, resulting in the up-regulation of the expression of the microphthalmia-associated transcription factor (MITF) which is a critical regulatory factor of melanin synthesis, and tyrosinase (TYR), the rate-limiting enzyme of melanin synthesis tyrosinase (TYR), and ultimately affects production of eumelanin and pheomelanin, and the coat color phenotype of mammalian species. Previous reports have indicated that the mutation A243T in the transmembrane domain 6 (TM6) of MC1R protein might disrupt the function of MC1R, contributing to the red phenotype in Duroc pig. However, functional analysis of the A243T mutation in MC1R has not yet been carried out. In this study, we attempted to used single-stranded oligo-deoxyribonucleotides (ssODN) as donor templates to introduce the c.727G>A (A243T) mutation into MC1R in human melanoma cell line SK-MEL-2 by CRISPR/Cas9 to analyze its effects on MC1R functions. We found the occurrence of ssODN recombination reached to 10%. Unfortunately, Sanger sequencing MC1R in six single-cell clones revealed that none carried the c.727G>A mutation, but all carried undesired mutations surrounding the target site. Cells transfected with CRISPR/Cas9 plasmids and ssODN presented significantly attenuated cAMP activation, and down-regulated MITF and TYR expression, indicating that the editing MC1R could affect the melanin synthesis function in cells. This study provides a basis for further investigation the mechanism of MC1R mutation on animal coat color.

Editing the cystine knot motif of MSTN enhances muscle development of Liang Guang Small Spotted pigs.

Myostatin (MSTN) is a member of the transforming growth factor-ß (TGF-ß) family, and functions as an inhibitor of muscle growth. Disrupting the inhibitory effect of MSTN on growth can provide an effective way to increase the muscle yield of livestock and poultry. The cysteine knot motif of TGF-ß can stabilize the structure of MSTN protein and plays an important regulatory role in the biological function of MSTN. Accordingly, in this study, we used the CRISRP/Cas9 to edit the exon 3 of MSTN in the kidney cells of Liang Guang Small Spotted pig (LPKCs), in order to disrupt the cysteine knot motif of MSTN and remove the inhibitory effect of MSTN on its target genes.MSTN-edited LPKCs were obtained through fluorescence-activated cell sorting (FACS) and used as donor cells for somatic cell nuclear transfer (SCNT) to generate cloned embryos, which were then transferred to surrogate sows to finally obtain eight MSTN-edited Liang Guang Small Spotted piglets. Among them, two survived to 10 days old. Genotyping revealed that these two piglets were gene edited heterozygotes with base deletion and substitution occurred within the coding sequence of C106 and C108 at the cystine knot motif of MSTN. These changes resulted in frameshift mutations, and conversion of C106 and C108 to other amino acids. More developments of muscles were observed at the shoulders and hips of the heterozygotes of MSTN-edited Liang Guang Small Spotted pigs. H&E analysis showed that the cross-sectional area (CSA) of myofiber inMSTN-edited pigs was significantly decreased, and the number of myofiber were significantly increased. Western blot analysis showed that the disruption of C106 and C108 did not affect the expression of MSTN protein, but significantly up-regulated the expression of its target genes such as Myf5, MyoD, Myogenin and other myogenic regulatory factors. In summary, the gene-edited pig model obtained in this study did not cause complete loss of MSTN expression, and could retain other biological functions of MSTN, thereby promoting muscle growth while minimizing the potential adverse effects on complete loss of MSTN in the Liang Guang Small Spotted pigs.

Harvesting of surface plasmon polaritons: Role of the confinement factor.

Surface plasmon polaritons (SPPs) are propagating waves generated at the interface of a metal (metamaterial) and a dielectric. The intensity of SPPs often exponentially decays away from the surface, while their wavelengths can be tuned by the confinement effect. We present here a computational method based on quantum-mechanical theory to fully describe the interaction between confined SPPs and adsorbed molecules at the interface. Special attention has been paid to the roles of the confinement factor. Taking a prototype dye sensitized solar cell as an example, calculated results reveal that with the increase in the confinement factor in metal/dielectric interfaces, the breakdown of the conventional dipole approximation emerges, which allows efficient harvesting of SPPs with low excitation energies and, thus, increases the efficiency of the solar energy conversion by dye molecules. Furthermore, at the metamaterial/dielectric interface, SPPs with large confinement factors could directly excite the dye molecule from its ground singlet state to the triplet state, opening an entirely new channel with long-living carriers for the photovoltaic conversion. Our results not only provide a rigorous theory for the SPP-molecule interaction but also highlight the important role played by the momentum of the light in plasmon related studies.

[Generation of cell strains containing point mutations in HPRT1 by CRISPR/Cas9].

Mutations in Hypoxanthine-guanine Phosphoribosyltransferase1 (HPRT1) gene can lead to metabolic disorder of hypoxanthine and guanine metabolism, and other severe symptoms such as hypophrenia, gout, and kidney stones, called the Lesch-Nyhan disease (LND). Although the mutations are widely distributed throughout the HPRT1 gene, there are some isolated hot spots. In this study, we aim to introduce two previously reported hot spots, c.508 C>T and c.151 C>T, which could lead to premature translational termination in HPRT1 gene. Through CRISPR/Cas9 mediated homology-directed repair (HDR) by using single-stranded oligo-deoxyribonucleotides (ssODN) as donor template, we obtained cell clones containing these two mutations in HEK293T or HeLa cells. Targeted mutation of c.508 C>T and c.151 C>T reached to 16.3% and 10%, respectively. We further detect HPRT1 protein levels with Western blot and enzyme activity with 6-TG in 5 different cell clones. HPRT1 protein and its enzymatic activity both was hardly detected in homozygous mutant cells, while reduced HPRT1 protein expression and enzymatic activity was detected in heterozygous mutant cells. Our study will be beneficial to those who working on generation of cell or animal models of HRPT1 mutations, and provides a basis for further investigations on the genetic mechanism of Lesch-Nyhan disease.

Improving gene targeting efficiency on the porcine BMP15 gene mediated by CRISPR/Cas9 by using the RGS surrogate reporter system.

As Chinese have raised most pigs and consumed most pig products in the world, improving the fertility of sow is of economic benefits to the pig industry in China. The sheep BMP15 (bone morphogenetic protein 15) gene has been identified as a major gene for controlling ovulation rates and prolific traits, which are key factors affecting the fertility of livestock. As similar natural occurring mutations in the porcine BMP15 gene have not yet been reported, we speculated that introducing the same prolific sheep mutations into the porcine BMP15 gene by using the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) system.

Optical Excitation in Donor-Pt-Acceptor Complexes: Role of the Structure.

The optical properties of the Pt complexes in the form of donor-metal-acceptor (D-M-A) were studied at the first-principles level. Calculated results show that for the frontier molecular orbitals (MOs) of a D-M-A structure the energies of unoccupied frontier MO can be mainly determined by the interaction between M and A, whereas the M-A and M-D interactions both determine the energies of occupied frontier MO. By developing a straightforward transition dipole decomposition method, we found that not only the local excitations in D but also those in A can significantly contribute to the charge-transfer (CT) excitation. Furthermore, the calculations also demonstrate that by tuning the dihedral angle between D and A the transition probability can be precisely controlled so as to broaden the spectrum region of photoabsorption. For the D-M-A molecule with a delocalized π system in A, the CT excitation barely affects the electronic structures of metal, suggesting that the oxidation state of the metal can be kept during the excitation. These understandings for the optical properties of the D-M-A molecule would be useful for the design of dye-sensitized solar cells, photocatalysis, and luminescence systems.

Infrared spectra of small anionic water clusters from density functional theory and wavefunction theory calculations.

We performed systematic theoretical studies on small anionic water/deuterated water clusters W/D(-)(N=2-6) at both density functional theory (B3LYP) and wavefunction theory (MP2) levels. The focus of the study is to examine the convergence of calculated infrared (IR) spectra with respect to the increasing number of diffuse functions. It is found that at the MP2 level for larger clusters (n = 4-6), only one extra diffuse function is needed to obtain the converged relative IR intensities, while two or three more sets of extra diffuse functions are needed for smaller clusters. Such behaviour is strongly associated with the convergence of the electronic structure of corresponding clusters at the MP2 level. It is striking to observe that at the B3LYP level, the calculated relative IR intensities for all the clusters under investigations are diverse and show no trend of convergence upon increasing the number of diffuse functions. Moreover, the increasing contribution from the extra diffuse functions to the dynamic IR dipole moment indicates that the B3LYP electronic structure also fails to converge. These results manifest that MP2 is a preferential theoretical method, as compared to the widely used B3LYP, for the IR intensity of dipole bounded electron systems.

Improving gene targeting efficiency on pig IGF2 mediated by ZFNs and CRISPR/Cas9 by using SSA reporter system.

IGF2 (Insulin-like growth factor 2) is a major growth factor affecting porcine fetal and postnatal development. We propose that the precise modification of IGF2 gene of Chinese indigenous pig breed--Lantang pig by genome editing technology could reduce its backfat thickness, and increase its lean meat content. Here, we tested the genome editing activities of zinc finger nucleases (ZFNs) and CRISPR/Cas9 system on IGF2 gene in the Lantang porcine fetal fibroblasts (PEF). The results indicated that CRISPR/Cas9 presented cutting efficiency up to 9.2%, which was significantly higher than that generated by ZFNs with DNA cutting efficiency lower than 1%. However, even by using CRISPR/Cas9, the relatively lower percentage of genetically modified cells in the transfected population was not satisfied for somatic nuclear transfer (SCNT). Therefore, we used a SSA (Single-strand annealing) reporter system to enrich genetically modified cells induced by ZFN or CRISPR/Cas9. T7 endonuclease I assay revealed that this strategy improved genome editing activity of CRISPR/Cas9 by 5 folds, and was even more effective for improving genome editing efficiency of ZFN.

[Spatial and Temporal Distribution Characteristics of Ozone Concentration and Health Benefit Assessment in the Beijing-Tianjin-Hebei Region from 2015 to 2020].

To evaluate the spatial and temporal distribution characteristics of ambient ozone (O3) in the Beijing-Tianjin-Hebei (BTH) Region, the land use regression (LUR) model and random forest (RF) model were used to simulate the ambient O3 concentration from 2015 to 2020. Meanwhile, all-cause, cardiovascular, and respiratory mortalities as well as economic losses attributed to O3 were also estimated. The results showed that upward trends with fluctuation were observed for ambient O3 concentration, mortalities, and economic losses attributable to O3 exposure in the BTH Region from 2015 to 2020. The areas with high O3 concentration and great changes were concentrated in the central and southwestern regions, whereas the concentration in the northern region was low, and the change degree was small. The spatial distribution of the mortalities was also consistent with the spatial distribution of O3 concentration. From 2015 to 2020, the economic losses regarding all-cause mortality and cardiovascular mortality increased in 13 cities of the BTH Region, whereas the economic losses of respiratory mortality decreased in 4 cities in the BTH Region. The results indicated that the priority areas for O3 control were not uniform. Specifically, Beijing, Tianjin, Hengshui, and Xingtai were vital areas for O3 pollution control in the BTH Region. Differentiated control measures should be adopted based on the characteristics of these target areas to decline O3 concentration and reduce health impacts and economic losses associated with O3 exposure.

Effects of mind-regulating acupuncture on pain, negative emotion and sleep quality in patients with postherpetic neuralgia： a randomized controlled pilot trial. / è°ƒç¥žé’ˆæ³•å¯¹å¸¦çŠ¶ç–±ç–¹åŽé—ç¥žç»ç—›æ‚£è€ ç–¼ç—›ç¨‹åº¦ã€è´Ÿæ€§æƒ ç»ªå’Œç¡çœ è´¨é‡çš„å½±å“.

OBJECTIVES: To observe the effect of mind-regulating acupuncture on pain intensity, sleep quality, negative emotion in patients with postherpetic neuralgia (PHN), and evaluate the clinical effect of mind-regulating acupuncture combined with surrounding needling and heavy moxibustion at Ashi points (Extra) in treatment of PHN. METHODS: The patients with PHN were randomly divided into a control group (35 cases, 2 cases dropped out) and a comprehensive therapy group (35 cases). The patients in the control group were treated with surrounding needling and heavy moxibustion at Ashi points. In the comprehensive therapy group, the mind-regulating acupuncture therapy was delivered besides the treatment as the control group. The treatment was given once daily, one course of treatment was composed of 6 days and 2 courses were required in the 2 groups. Before and after treatment, the pain conditions were assessed using pain rating index (PRI), visual analogue scale (VAS) and present pain intensity (PPI), the negative emotions were assessed using Hamilton anxiety scale (HAMA) and Hamilton depression scale (HAMD), and the sleep quality with Pittsburgh sleep quality index (PSQI). One week before and one week after treatment, the average sleep time was recorded. The therapeutic effect of 2 groups was evaluated. The effective cases of 2 groups were followed up in 2 months after treatment completion and the recurrence of neuralgia was recorded. RESULTS: There were no statistical differences in the above indicators between the 2 groups before treatment. After 2 courses of treatment, the scores of PRI, VAS, PPI, HAMA, HAMD and PSQI were reduced when compared with those before treatment in the patients of the 2 groups (P<0.05), and the average sleep time was increased (P<0.05). The scores of PRI, VAS, PPI, HAMA, HAMD and PSQI in the comprehensive therapy group, as well as the average sleep time were all improved when compared with those of the control group (P<0.05). The total effective rate in the comprehensive therapy group (34/35, 97.14%) was higher than that of the control group (27/33, 81.82%, P<0.05) and the recurrence rate was lower (ï¼»2/34, 5.88%ï¼½vsï¼»8/27, 29.63%ï¼½, P<0.05). CONCLUSIONS: The combination of mind-regulating acupuncture with surrounding needling and heavy moxibustion at Ashi acupoint can effectively relieve PHN. Compared with the traditional surrounding acupuncture in pain area combined with moxibustion at Ashi points, this comprehensive therapy is more effective for ameliorating pain intensity, improving sleep quality and reducing negative emotions. It is also effective for declining the recurrence.

[Effects of Rosa roxburghii Pomace Biochar on Yield and Quality of Chinese Cabbage and Soil Properties].

In order to explore the effect of Rosa roxburghii pomace biochar on the yield and quality of Chinese cabbage and soil properties and realize the resource utilization of R. roxburghii pomace, a pot experiment was conducted to study the effect of R. roxburghii pomace biochar on the yield and quality of Chinese cabbage and soil properties by setting five biochar application rates of 0 % (CK), 1 % (T1), 3 % (T2), 5 % (T3), and 7 % (T4). The results showed that:① The application of R. roxburghii pomace biochar could significantly improve the yield and quality of Chinese cabbage, and the effect was the best at a 5 % biochar application rate. The yield, soluble solids, soluble sugar, vitamin C, total nitrogen, total phosphorus, and total potassium content of Chinese cabbage increased by 71.51 %, 40.14 %, 33.65 %, 38.08 %, 9.03 %, 28.85 %, and 35.38 %, respectively, compared with those in CK. ② The application of biochar from R. roxburghii pomace could significantly improve soil properties and increase soil nutrient content and availability. The effect was better at a 5 % biochar application rate. The soil pH, organic matter, total nitrogen, alkali-hydrolyzable nitrogen, available phosphorus, and available potassium content increased by 41.06 %, 134.84 %, 157.48 %, 140.79 %, 341.75 %, and 627.13 %, respectively, compared with those in CK. The contents of available Fe, Mn, Cu, and Zn and exchangeable Ca and Mg increased by 37.68 %, 61.69 %, 400.00 %, 4 648.84 %, 617.17 %, and 351.42 %, respectively, compared with those in CK. ③ The application of biochar from R. roxburghii pomace could significantly enhance soil enzyme activity. Compared with those in the CK treatment, soil urease, acid phosphatase, catalase, and sucrase increased by 51.43 %-362.86 %, 90.63 %-134.14 %, 21.40 %-85.12 %, and 82.92 %-218.43 %, respectively. ④ Redundancy analysis showed that soil AK; exchangeable Ca, SOM, and AP; and available Zn were the main factors affecting the yield and quality of Chinese cabbage, and there was a significant positive correlation between them. In summary, the application of R. roxburghii pomace biochar can significantly increase the yield and quality of Chinese cabbage and improve soil properties. The preparation of R. roxburghii pomace into biochar can provide a theoretical reference for the rational utilization of R. roxburghii pomace resources.

Ginkgolide B reduces neuronal cell apoptosis in the traumatic rat brain: possible involvement of toll-like receptor 4 and nuclear factor kappa B pathway.

Ginkgolide B (GB) has been demonstrated to have a variety of pharmacological actions. Accumulating evidence indicates that GB may exert a protective effect on brain injury. The study was designed to investigate the influence of GB on toll-like receptor 4 (TLR-4) and nuclear factor κB (NF-κB)-dependent inflammatory responses and neuronal cell apoptosis after traumatic brain injury (TBI). Wistar rats were subjected to 5, 10 and 20 mg/kg GB daily for 5 days, intraperitoneally, following TBI. Rats were sacrificed at hour 2, 6 and 12, as well as day 1, 2, 3 and 5 after TBI. The administration of 10 and 20 mg/kg GB could significantly (least-significant difference test: p < 0.05) suppress gene expressions of TLR-4 and NF-κB, lessen concentrations of tumour necrosis factor α, interleukin-1ß and interleukin-6, as well as reduce the number of apoptotic neuronal cells in traumatic rat brain tissues, but the administration of 5 mg/kg GB did not (p > 0.05). However, a clear concentration-response relationship was not found. Thus, GB may inhibit TLR-4 and NF-κB-dependent inflammatory responses, and furthermore lessen neuronal cell apoptosis after TBI, which may support the use of GB for the treatment of TBI.

Optical Images of Molecular Vibronic Couplings from Tip-Enhanced Fluorescence Excitation Spectroscopy.

Tip-based photoemission spectroscopic techniques have now achieved subnanometer resolution that allows visualization of the chemical structure and even the ground-state vibrational modes of a single molecule. However, the ability to visualize the interplay between electronic and nuclear motions of excited states, i.e., vibronic couplings, is yet to be explored. Herein, we theoretically propose a new technique, namely, tip-enhanced fluorescence excitation (TEFE). TEFE takes advantage of the highly confined plasmonic field and thus can offer a possibility to directly visualize the vibronic effect of a single molecule in real space for arbitrary excited states in a given energy window. Numerical simulations for a single porphine molecule confirm that vibronic couplings originating from Herzberg-Teller (HT) active modes can be visually identified. TEFE further enables high-order vibrational transitions that are normally suppressed in the other plasmon-based processes. Images of the combination vibrational transitions have the same pattern as that of their parental HT active mode's fundamental transition, providing a direct protocol for measurements of the activity of Franck-Condon modes of selected excited states. These findings strongly suggest that TEFE is a powerful strategy to identify the involvement of molecular moieties in the complicated electron-nuclear interactions of the excited states at the single-molecule level.

Oxymatrine reduces neuronal cell apoptosis by inhibiting Toll-like receptor 4/nuclear factor kappa-B-dependent inflammatory responses in traumatic rat brain injury.

OBJECTIVE: To investigate the influence of oxymatrine (OMT) on Toll-like receptor 4 (TLR-4)/nuclear factor kappa-B (NF-κB)-dependent inflammatory responses and neuronal cell apoptosis after traumatic brain injury (TBI). MATERIALS AND METHODS: Wistar rats were given an intraperitoneal injection of 60 or 120 mg/kg OMT after TBI once a day till day 5. Rats were killed by decapitation at hours 2, 6 and 12, and days 1, 2, 3 and 5 after TBI. Gene expressions of TLR-4 and NF-κB, concentrations of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1ß) and interleukin-6 (IL-6) as well as the number of apoptotic neuronal cells in traumatic rat brain tissues were determined. RESULTS: The administration of 120 mg/kg OMT could significantly suppress gene expressions of TLR-4 and NF-κB, lessen concentrations of TNF-α, IL-1ß and IL-6, and reduce the number of apoptotic neuronal cells in traumatic rat brain tissues by the Mann-Whitney U test (P < 0.05), but the administration of 60 mg/kg OMT could not (P > 0.05). CONCLUSION: OMT may inhibit TLR4/NF-κB-dependent inflammatory responses, and furthermore lessen neuronal cell apoptosis after TBI.

Copeptin is associated with mortality in patients with traumatic brain injury.

BACKGROUND: High serum copeptin levels are associated with injury severity after traumatic brain injury (TBI). However, not much is known regarding its relation with mortality. Thus, we sought to evaluate its relation with disease mortality. METHODS: Fifty healthy controls and 94 patients with acute severe TBI were included. Plasma samples were obtained on admission and at days 1, 2, 3, 5, and 7. Its concentration was measured by enzyme-linked immunosorbent assay. RESULTS: Twenty-six patients (27.7%) died from TBI in a month. After brain injury, plasma copeptin level in patients increased during the 6-hour period immediately, peaked in 24 hours, plateaued at day 2, decreased gradually thereafter, and was substantially higher than that in healthy controls during the 7-day period. A forward stepwise logistic regression selected plasma copeptin level (odds ratio, 1.008; 95% confidence interval, 1.002-1.014; p = 0.010) as an independent predictor for 1-month mortality of patients. A multivariate linear regression showed that plasma copeptin level was negatively associated with Glasgow Coma Scale (GCS) score (t = -7.161; p < 0.001). A receiver operating characteristic curve identified plasma copeptin cutoff level (451.8 pg/mL) that predicted 1-month mortality with the optimal sensitivity (88.5%) and specificity (75.0%) values (area under curve, 0.874; 95% confidence interval, 0.789-0.933; p < 0.001). The area under curve of plasma copeptin level was similar to that of GCS score (p = 0.299). However, copeptin did not statistically significantly improve the area under curve of GCS score (p = 0.413). CONCLUSIONS: Increased plasma copeptin levels are associated with mortality after TBI.

Usefulness of serum interleukin-33 as a prognostic marker of severe traumatic brain injury.

BACKGROUND: Interleukin-33 is recently identified as a brain injury biomarker. We determined whether serum interlerukin-33 concentrations are associated with inflammation, severity and prognosis after traumatic brain injury (TBI). METHODS: We detected serum interlerukin-33 concentrations of 102 healthy controls and 102 severe TBI patients, as well as serum concentrations of 3 inflammatory biomarkers (interleukin-6, tumor necrosis factor-alpha and C-reactive protein) and 7 cell-specific proteins (myelin basic protein, glial fibrillary astrocyte protein, S100B, neuron-specific enolase, phosphorylated axonal neurofilament subunit H, Tau and ubiquitin carboxyl-terminal hydrolase L1) in 102 severe TBI patients. The recorded poor prognosis variables included acute lung injury, acute traumatic coagulopathy, progressive hemorrhagic injury, posttraumatic cerebral infarction and six-month mortality and poor outcome (Glasgow score of 1-3). RESULTS: Median interlerukin-33 concentration of patients (692 pg/mL) was substantially raised, as compared to controls. Interlerukin-33 concentrations were significantly correlated with Glasgow coma scale (GCS) score and the preceding biomarkers concentrations. Interlerukin-33 concentration > 692 pg/mL emerged as an independent prognostic predictor and its discriminatory capability exceeded those of the above-mentioned inflammatory biomarkers concentrations and was in the range of GCS scores and the aforementioned cell-specific proteins concentrations. CONCLUSION: Ascending serum interlerukin-33 concentrations could reflect inflammation, severity and worse prognosis following TBI.

MiR-9 Promotes Apoptosis Via Suppressing SMC1A Expression in GBM Cell Lines.

OBJECTIVE: Glioblastomas multiforme (GBM) is the most malignant brain cancer, which presented vast genomic variation with complicated pathologic mechanism. METHOD: MicroRNA is a delicate post-transcriptional tuner of gene expression in the organisms by targeting and regulating protein coding genes. MiR-9 was reported as a significant biomarker for GBM patient prognosis and a key factor in regulation of GBM cancer stem cells. To explore the effect of miR-9 on GBM cell growth, we over expressed miR-9 in U87 and U251 cells. The cell viability decreased and apoptosis increased after miR-9 overexpression in these cells. To identify the target of miR-9, we scanned miR-9 binding site in the 3'UTRs region of expression SMC1A (structural maintenance of chromosomes 1A) genes and designed a fluorescent reporter assay to measure miR-9 binding to this region. Our results revealed that miR-9 binds to the 3'sUTR region of SMC1A and down-regulated SMC1A expression. RESULT: Our results indicated that miR-9 was a potential therapeutic target for GBM through triggering apoptosis of cancer cells.

Serum thioredoxin and in-hospital major adverse events after traumatic brain injury.

BACKGROUND: In-hospital major adverse events (IMAEs), mainly including acute lung injury, acute traumatic coagulopathy, progressive hemorrhagic injury and posttraumatic cerebral infarction, are associated with poor prognosis after traumatic brain injury (TBI). Thioredoxin, a potent anti-oxidant, has been identified as an oxidative stress marker. This study was designed to explore the association of serum thioredoxin concentrations with IMAEs of patients with severe TBI. METHODS: This prospective, observational study recruited a total of 108 healthy controls and 108 patients with severe TBI. We investigated the possible relation of serum thioredoxin concentrations to IMAEs and trauma severity (reflected by Glasgow coma scale scores) following TBI using a multivariate analysis. RESULTS: Serum thioredoxin concentrations were higher in the patients than in the controls. Serum concentrations of thioredoxin significantly correlated with admission Glasgow coma scale scores. Thioredoxin in serum independently predicted any IMAEs. As compared to admission Glasgow coma scale scores, thioredoxin concentrations had similar areas under receiver operating characteristic curve for any IMAEs. CONCLUSION: Increased serum thioredoxin concentrations are highly associated with trauma severity and IMAEs, indicating thioredoxin might be a potential prognostic biomarker after TBI.

Serum macrophage migration inhibitory factor concentrations correlate with prognosis of traumatic brain injury.

BACKGROUND: Macrophage migration inhibitory factor (MIF) is a well-known pro-inflammatory cytokine. Serum MIF concentrations are associated with the severity and prognosis of ischemic stroke. METHODS: In this prospective, observational study, white blood cell (WBC) count and serum concentrations of C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and MIF among 108 severe traumatic brain injury (TBI) patients and 108 controls were measured. We determined whether serum MIF concentrations are associated with inflammation, severity, in-hospital major adverse events (IMAEs) (i.e., in-hospital mortality, acute lung injury, acute traumatic coagulopathy, progressive hemorrhagic injury and posttraumatic cerebral infarction) and long-term clinical outcome (i.e., 6-month functional outcome) after TBI. RESULTS: As compared to the controls, serum CRP, IL-6, TNF-α and MIF concentrations were significantly increased. MIF concentrations correlated with WBC count, CRP, IL-6 and TNF-α concentrations and Glasgow coma scale (GCS) scores. MIF in serum was independently associated with IMAEs and long-term clinical outcome. Area under receiver operating characteristic curve of MIF concentrations was similar to GCS scores'. Moreover, MIF concentrations markedly improved the predictive value of GCS scores for 6-month unfavorable outcome. CONCLUSION: Increased serum MIF concentrations have close relation to inflammation, trauma severity and clinical outcomes, substantializing MIF as a good prognostic biomarker after TBI.