Dual regulation effect and mechanism of human myeloid-derived suppressor cells on anticolorectal cancer cells activity of Vγ9Vδ2 T cells.

Myeloid-derived suppressor cells (MDSC) are a group of immature inhibitory cells of bone marrow origin. Human γδ T cells (mainly Vγ9Vδ2 T cells) have emerged as dominant candidates for cancer immunotherapy because of their unique recognition pattern and broad killing activity against tumor cells. Intestinal mucosal intraepithelial lymphocytes are almost exclusively γδ T cells, so it plays an important role in inhibiting the development of colorectal cancer. In this study, we investigated the effects and molecular mechanism of human MDSC on anticolorectal cancer cells activity of Vγ9Vδ2 T cells. Our results suggested that MDSC can reduce the NKG2D expression of Vγ9Vδ2 T cells through direct cell-cell contact, which is associated with membrane-type transforming growth factor-ß. In contrast, MDSC can increase Vγ9Vδ2 T cells activation and production of IFN-γ, perforin, Granzyme B through direct cell-cell contact. This may be related to the upregulation of T-bet in Vγ9Vδ2 T cells by MDSC. However, MDSC had a dominant negative regulatory effect on the anticolorectal cancer cells activity of Vγ9Vδ2 T cells. Our study provides a theoretical basis for the immune regulatory function of human MDSC on γδ T cells. This will be conducive to the clinical development of a new antitumor therapy strategy.

Low-intensity transcranial ultrasound stimulation modulates neural activities in mice under propofol anaesthesia.

BACKGROUND: Previous studies have reported that transcranial focused ultrasound stimulation can significantly decrease the time to emergence from intraperitoneal ketamine-xylazine anaesthesia in rats. However, how transcranial focused ultrasound stimulation modulates neural activity in anaesthetized rats is unclear. METHODS: In this study, to answer this question, we used low-intensity transcranial ultrasound stimulation (TUS) to stimulate the brain tissue of propofol-anaesthetized mice, recorded local field potentials (LFPs) in the mouse motor cortex and electromyography (EMG) signals from the mouse neck, and analysed the emergence and recovery time, mean absolute power, relative power and entropy of local field potentials. RESULTS: We found that the time to emergence from anaesthesia in the TUS group (20.3 ± 1.7 min) was significantly less than that in the Sham group (32 ± 2.6 min). We also found that compared with the Sham group, 20 min after low-intensity TUS during recovery from anaesthesia, (1) the absolute power of local field potentials in mice was significantly reduced in the [1-4 Hz] and [13-30 Hz] frequency bands and significantly increased in the [55-100 Hz], [100-140 Hz] and [140-200 Hz] frequency bands; (2) the relative power of local field potentials in mice was enhanced at [30-45 Hz], [100-140 Hz] and [140-200 Hz] frequency bands; (3) the entropy of local field potentials ([1-200 Hz]) was increased. CONCLUSION: These results demonstrate that low-intensity TUS can effectively modulate neural activities in both awake and anaesthetized mice and has a positive effect on recovery from propofol anaesthesia in mice.

Revisiting the relationship between complement and ulcerative colitis.

Ulcerative colitis (UC) is an inflammatory bowel disorder (IBD) characterized by relapsing chronic inflammation of the colon that causes continuous mucosal inflammation. The global incidence of UC is steadily increasing. Immune mechanisms are involved in the pathogenesis of UC, of which complement is shown to play a critical role by inducing local chronic inflammatory responses that promote tissue damage. However, the function of various complement components in the development of UC is complex and even paradoxical. Some components (e.g. C1q, CD46, CD55, CD59, and C6) are shown to safeguard the intestinal barrier and reduce intestinal inflammation, while others (e.g. C3, C5, C5a) can exacerbate intestinal damage and accelerate the development of UC. The complement system was originally thought to function primarily in an extracellular mode; however, recent evidence indicates that it can also act intracellularly as the complosome. The current study provides an overview of current studies on complement and its role in the development of UC. While there are few studies that describe how intracellular complement contributes to UC, we discuss potential future directions based on related publications. We also highlight novel methods that target complement for IBD treatment.

3-Methyladenine ameliorates surgery-induced anxiety-like behaviors in aged mice by inhibiting autophagy-induced excessive oxidative stress.

BACKGROUND: Postoperative anxiety is a common surgical complication in older patients. Research has recently linked excessive autophagy to several neurological disorders, including anxiety. This study aimed to determine whether 3-Methyladenine (3-MA) administration reduced anxiety-like behaviors in a mouse model following abdominal exploratory laparotomy. METHODS: An abdominal exploratory laparotomy model of postoperative anxiety was established using male C57BL/6 mice aged 20 months. 3-MA (6, 30, and 150 mg/ml) was administered via intracerebroventricular immediately following surgery. The mice were assessed 14 days after surgery using the marble burying, elevated plus maze tests, and local field potential recording in the amygdala. The levels of expression of phosphorylated-Akt, Beclin-1, LC3B, nuclear factor erythroid 2-related factor 2 (Nrf2)-occupied regions in NeuN-positive cells, superoxide dismutase (SOD) activity, malondialdehyde (MDA), and glutathione (GSH) were measured at 24 h after surgery. RESULTS: The injection of 3-MA reversed the increased number of marbles buried, decreased time spent in the open arm, and enhanced θ oscillation power after 14 days of abdominal exploratory laparotomy. In addition, administration of 3-MA reduced the ratio of phosphorylated- to total-Akt, decreased expression in Beclin-1 and LC3B, attenuated MDA levels, and increased the ratio of Nrf2-occupied areas in NeuN-positive cells, SOD activity, and GSH levels under abdominal exploratory laparotomy conditions. CONCLUSIONS: 3-MA improved anxiety-like behaviors in aged mice undergoing abdominal exploratory laparotomy by inhibiting excessive autophagy-induced oxidative stress. These results suggest that 3-MA could be an effective treatment for postoperative anxiety.

Comparison of the accuracy of three methods measured the length of the right main stem bronchus by chest computed tomography as a guide to the use of right sided double-lumen tube.

BACKGROUND: The variation of right main stem bronchus leads to the orifice of the right upper lobe bronchus may be obstructed or increase the incidence of malposition intraoperatively when the right sided double-lumen tube is used. Therefore, the aim of this study was to compare the accuracy of three methods measured the length of the right main stem bronchus via chest computed tomography as a guide to the use of right sided double-lumen tube. METHODS: In this study, 168 adult patients undergoing left sided thoracic surgery were included. All these patients were allocated to carina-proximal (C-P) group, carina-distal (C-D) group and carina-carina (C-C) group. The position of endobronchial cuff observed via Fiberoptic bronchoscopy after successful initial placement and after turning the patients to the lateral decubitus position, as well as the incidence of malposition of right sided double-lumen tube intraoperative were recorded to assess the accuracy of three methods in predicting the position of right sided double-lumen tube. RESULTS: The distance between the carina to the proximal margin of the right upper lobe orifice, carina to the distal margin of the right upper lobe orifice and carina to the first right interlobar carina of the right upper lobe orifice were 17.2 ± 2.3 mm, 25.4 ± 3.7 mm and 28.5 ± 3.1 mm (P < 0.05). In the C-D group, the number of endobronchial cuffs seen to be herniating out of the carina, the number of bronchoscopies during initial placement and on the lateral position, the number of total malposition intraoperative and the number of reposition manoeuvres intraoperative were significantly less than the C-P group or the C-C group (P < 0.05). CONCLUSIONS: The length of the right main stem bronchus measured by the carina to distal margin of right upper lobe orifice method was more accurate than the other two methods in guiding the use of right sided double-lumen tube. TRIALS REGISTRATION: Clinical Trials. gov. no. NCT04127903. Registered at https://register. CLINICALTRIALS: gov on 16/10/2019.

Fc-specific and covalent conjugation of a fluorescent protein to a native antibody through a photoconjugation strategy for fabrication of a novel photostable fluorescent antibody.

Fluorophore-antibody conjugates with high photobleaching resistance, high chemical stability, and Fc-specific attachment is a great advantage for immunofluorescence imaging. Here, an Fc-binding protein (Z-domain) carrying a photo-cross-linker (p-benzoylphenylalanine, Bpa) fused with enhanced green fluorescent protein (EGFP), namely photoactivatable ZBpa-EGFP recombinant, was directly generated using the aminoacyl-tRNA synthetase/suppressor tRNA technique without any further modification. By employing the photoactivatable ZBpa-EGFP, an optimal approach was successfully developed which enabled EGFP to site-selectively and covalently attach to native antibody (IgG) with approximately 90% conjugation efficiency. After characterizing the Fc-specific and covalent manner of the EGFP-photoconjugated antibody, its excellent photobleaching resistance for immunofluorescence imaging was demonstrated in a model study by monitoring the toll-like receptor 4 (TLR4) expression in HepG2 cells. The proposed approach here for the preparation of a novel fluorescent antibody is available and reliable, which would play an important role in fluorescence immunoassay, and is expected to be extended to the generation of other biomolecule-photoconjugated antibodies, such as other fluorescent proteins for multiplex immunofluorescence imaging or reporter enzymes for highly sensitive enzyme immunoassays.Graphical abstract.

Effects of head positions on awake fiberoptic bronchoscope oral intubation: a randomized controlled trial.

BACKGROUND: There are many factors affecting the success rate of awake orotracheal intubation via fiberoptic bronchoscope. We performed this study was to investigate the effects of head positions on awake Fiberoptic bronchoscope oral intubation. METHODS: Seventy-five adult patients, received general anaesthesia were included in this study. After written informed consent, these patients were undergoing awake orotracheal intubation via fiberoptic-bronchoscope and according to the head position, the patients were randomized allocated to neutral position group (NP group), sniffing position group (SP group) or extension position group (EP group). After sedation the patients were intubated by an experienced anesthesiologist. The time to view the vocal cords, the percentage of glottic opening scores (POGO), the time to insert the tracheal tube into trachea and the visual analog scale (VAS) scores for ease experienced of passing the tracheal tube through glottis, the hemodynamic changes and the adverse events after surgery were recorded. RESULTS: The time to view the vocal cords was significantly shorter and the POGO scores was significantly higher in the EP group compared with the other two groups (P < 0.05); The SpO2 in the EP group was higher than NP group at before intubation and higher than SP group and NP group at immediate after intubation (P < 0.05); The time to insert the tracheal tube into trachea, the VAS scores for passing the tracheal tube through glottis, the coughing scores had no significant differences among groups (P > 0.05). There were also no significant differences regard to the incidence of postoperative complications, mean arterial pressure and heart rate among the groups (P > 0.05). CONCLUSIONS: The head at extension position had a best view of glottic opening than neutral position or sniffing position during awake Fiberoptic bronchoscope oral intubation, so extension position was recommended as the starting head position for awake Fiberoptic bronchoscope oral intubation. TRIAL REGISTRATION: Clinical Trials.gov. no. NCT02792855. Registered at https://register.clinicaltrials.gov on 23 september 2017.

Leucine aminopeptidase 3 promotes migration and invasion of breast cancer cells through upregulation of fascin and matrix metalloproteinases-2/9 expression.

Overexpression of leucine aminopeptidase 3 (LAP3) is involved in proliferation, migration, and invasion of several tumor cells and plays a crucial role in tumor metastasis. However, the related mechanism remains unknown. In this study, we used MDA-MB-231 and MCF7 breast cancer cell lines to explore the role of LAP3 in the regulation of cancer cell migration and invasion by employing the natural LAP3 inhibitor bestatin and a lentivirus vector that overexpresses or knocks down LAP3. Bestatin inhibited tumor cell migration and invasion in a dose-dependent manner. Western blot assay showed that bestatin and knockdown of LAP3 upregulated phosphorylation of Hsp27 and downregulated expression of fascin. Phosphorylation of Akt and expression of matrix metalloproteinase-2/9 can also be downregulated. LAP3 overexpression showed the opposite results. Immunohistochemistry analysis was conducted to detect expression levels of LAP3 in breast cancer tissues. High LAP3 expression was correlated with the grade of malignancy. Findings of this study uncovered the molecular mechanism of LAP3 on breast cancer metastasis and indicated that LAP3 may act as a potential antimetastasis therapeutic target.

Response Surface Methodology for Optimizing the Ultrasound-Assisted Extraction of Polysaccharides from Acanthopanax giraldii.

The aim of this study was to investigate and optimize the most important factors affecting the extraction of Acanthopanax giraldii HARMS polysaccharides (AHPs) by ultrasound-assisted extraction (UAE) technology in a systemic manner. The ranges of four factors, including extraction temperature, liquid/solid ratio, extraction time, and ultrasonic power, were first determined by a single-factor experiment, followed by optimization of the UAE conditions using the Box-Behnken design (BBD) for maximum AHPs production. In our study, the models developed from the experimental design predicted the experimental data well and had a high determination coefficient (R2=0.9387). The optimized conditions for AHPs extraction were as follows: extraction temperature, 58°C; liquid/solid ratio, 25 : 1; extraction time, 73 min; and ultrasonic power, 85 W. Under these optimized conditions, the polysaccharide yield was 1.532±0.037% (n=3), being very close to the predicted value of 1.546% by the model. In addition, to investigate whether there was a difference of AHPs content between UAE and traditional hot water extraction (THWE), Fourier-transform (FT) IR spectral analyses was performed. The results showed that the functional groups of the polysaccharides extracted by either UAE or THWE were fundamentally identical. Furthermore, AHPs extracted by UAE could promote macrophage activation, such as enhanced phagocytosis and increased cytokine (interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α)) secretion in RAW264.7 cells. In conclusion, optimization of the UAE conditions by response surface methodology (RSM) was a promising method to improve the extraction yield of AHPs. AHPs extracted by the optimized UAE method can maintain their polysaccharide structure and biological activity.

Pre-B cell leukemia homeobox 1 is associated with lupus susceptibility in mice and humans.

Sle1a.1 is part of the Sle1 susceptibility locus, which has the strongest association with lupus nephritis in the NZM2410 mouse model. In this study, we show that Sle1a.1 results in the production of activated and autoreactive CD4(+) T cells. Additionally, Sle1a.1 expression reduces the peripheral regulatory T cell pool, as well as induces a defective response of CD4(+) T cells to the retinoic acid expansion of TGF-ß-induced regulatory T cells. At the molecular level, Sle1a.1 corresponds to an increased expression of a novel splice isoform of Pbx1, Pbx1-d. Pbx1-d overexpression is sufficient to induce an activated/inflammatory phenotype in Jurkat T cells and to decrease their apoptotic response to retinoic acid. PBX1-d is expressed more frequently in the CD4(+) T cells from lupus patients than from healthy controls, and its presence correlates with an increased central memory T cell population. These findings indicate that Pbx1 is a novel lupus susceptibility gene that regulates T cell activation and tolerance.

PTTG1 promotes migration and invasion of human non-small cell lung cancer cells and is modulated by miR-186.

Deeper mechanistic understanding of non-small cell lung cancer (NSCLC), a leading cause of total cancer-related deaths, may facilitate the establishment of more effective therapeutic strategies. In this study, pituitary tumor transforming gene (PTTG1) expression was associated with lymph node and distant metastasis in patients with NSCLC and was correlated with patient survival. Reduction of PTTG1 by small interfering RNA (siRNA) inhibits the migration and invasion of NSCLC cells by mediating matrix metalloproteinases expression. To the best of our knowledge, this study is the first to report that PTTG1 promotes epidermal growth factor (EGF) induced the phosphorylation of LIN-11, Isl1 and MEC-3 protein domain kinase and cofilin, a critical step in cofilin recycling and actin polymerization. Additionally, EGF-induced Akt phosphorylation was suppressed through knockdown of PTTG1. Interestingly, miR-186 can modulate PTTG1 protein expression. As observed from the animal experiment in this study, knockdown of PTTG1 through siRNA and overexpression of miR-186 inhibited invasive activity of NSCLC cells toward the SCID mice lung. In summary, our in vitro and in vivo results indicate that PTTG1 modulated by miR-186 has an important function in NSCLC invasion/metastasis. This study identified both PTTG1 and miR-186 as potential anti-invasion targets for therapeutic intervention in NSCLC.

Immobilization of unraveled immunoglobulin G using well-oriented ZZ-His protein on functionalized microtiter plate for sensitive immunoassay.

Highly efficient protein immobilization is extremely crucial for solid-phase immunoassays. We present a strategy for oriented immobilization of functionally intact immunoglobulin G (IgG) on a polystyrene microtiter plate via iminodiacetic acid (IDA)-Ni(2+) and ZZ-His protein interaction. We immobilized a ZZ-EAP (Escherichia coli alkaline phosphatase)-His fusion protein, which exhibits Fc binding, His tag, and intrinsic AP activities, and analyzed it against the interaction between rabbit IgG anti-horseradish peroxidase (anti-HRP) and its binding partner HRP to investigate the specificity and efficacy of this method. We compared the IDA-Ni(2+)-(ZZ-His) method with ZZ-EAP random immobilization using sandwich enzyme-linked immunosorbent assay, and the results showed that the former method had an enhanced signal, 10-fold higher sensitivity, and a wider linear range. Thus, the proposed method allows a broad range of oriented immobilized functionally intact IgG antibodies on polystyrene plates using only one type of IDA-Ni(2+) chelate surface because the ZZ protein can bind to the Fc region of various IgGs.

Comparative characterization of recombinant ZZ protein-alkaline phosphatase and its application in enzyme immunoassays.

A functional fusion protein, which consists of an antibody and an enzyme that can be used in enzyme immunoassays, has been constructed. However, a quantitative comparison of the characteristics of fusion proteins and chemical conjugates of the parents, which are functionally produced in a uniform microbial system, has not been adequately achieved. In this study, a fusion protein between the ZZ protein and Escherichia coli alkaline phosphatase (AP) and the parental ZZ protein and AP for chemical conjugate was functionally produced in the same bacterial system. A detailed examination of the ZZ-AP fusion protein and the effect of the ZZ-AP chemical conjugate on IgG affinity and enzymatic activity were performed. Compared with the parents, the equilibrium dissociation constant of ZZ-AP conjugate decreased by 32 % and catalytic activity decreased by 24 %, whereas the ZZ-AP fusion retained full parental activities and exhibited an approximately tenfold higher sensitivity than that of ZZ-AP conjugate in enzyme-linked immunosorbent assay. Thus, ZZ-AP fusion is a promising immunoreagent for IgG detection and a potential biolinker between antibodies and reporter enzymes (i.e., IgG-ZZ-AP fusion complex) in immunoassays.

Appropriate tube temperature for fiberoptic bronchoscope-guided intubation of thermally softened double-lumen endotracheal tubes: A CONSORT-compliant article.

BACKGROUND: To compare the effects of thermal softening of double-lumen endotracheal tubes (DLT) at different temperatures during fiberoptic bronchoscopy (FOB)-guided intubation. METHODS: We randomly divided 144 patients undergoing thoracic surgery into 4 groups as follows: T1 (T = 24 ± 1°C, n = 36), T2 (T = 36 ± 1°C, n = 36), T3 (T = 40 ± 1°C, n = 36), and T4 (T = 48 ± 1°C, n = 36). All groups underwent FOB-guided double-lumen endotracheal intubation and positioning. We recorded the duration of positioning and intubation using DLT, intubation resistance (IR), the success rate of the first attempt at endotracheal intubation, and the incidence of postoperative vocal cord injury and hoarseness. RESULTS: The time to intubation was longer in the T1 group than that in the T2, T3, and T4 groups (P < .05). The time for positioning was longer in the T4 group than that in the T1, T2, and T3 groups (P < .05). IR was lower in the T3 and T4 groups than those in T1 and T2 groups (P < .05). The success rate of the first attempt at endotracheal intubation was higher in the T2, T3, and T4 groups than that in the T1 group (P < .05). Postoperative glottic injury and hoarseness were higher in the T1 and T2 groups than those in the T3 and T4 groups (P < .05). CONCLUSION: A thermally softened DLT shortened the time to intubation, reduced the IR, improved the success rate of the first attempt at endotracheal intubation, and lowered the incidence of postoperative glottic injury and hoarseness. The optimal tube temperature for FOB-guided intubation of thermally softened DLT was 40 ± 1°C.

Effect of Angelica polysaccharide on mouse myeloid-derived suppressor cells.

Angelica polysaccharide (APS) is a polysaccharide extracted from Angelica sinensis and it is one of the main active components of Angelica sinensis. Many studies have demonstrated that APS can promote the activation and function of a variety of immune cells and is recognized as an immune enhancer, but the regulatory effect of APS on myeloid-derived suppressor cells (MDSC) is still unclear. In this study, we investigated the effects of APS on MDSC proliferation, differentiation and function through in vivo and in vitro experiments. In vitro, our results showed that APS promoted the proliferation, differentiation and immunosuppressive function of MDSC through STAT1 and STAT3 signaling pathways, and positively correlated with the expression level of Mannose receptor (MR, also known as CD206) and in a concentration-dependent manner on APS. In vivo, APS up-regulated T cells, γδT cells, CD8+T cells, natural killer cells, monocytes/macrophages, and granulocytes in the peripheral blood and spleen of mice to varying degrees and was accompanied by the same degree of increase in the proportion of MDSC. That reminds to the clinician that when applying APS as treatment they should pay attention to its possible side effects of increasing the quantity and function of MDSC, in order to increase its efficacy.

NK Cells in Autoimmune Diseases: Protective or Pathogenic?

Autoimmune diseases generally result from the loss of self-tolerance (i.e., failure of the immune system to distinguish self from non-self), and are characterized by autoantibody production and hyperactivation of T cells, which leads to damage of specific or multiple organs. Thus, autoimmune diseases can be classified as organ-specific or systemic. Genetic and environmental factors contribute to the development of autoimmunity. Recent studies have demonstrated the contribution of innate immunity to the onset of autoimmune diseases. Natural killer (NK) cells, which are key components of the innate immune system, have been implicated in the development of multiple autoimmune diseases such as systemic lupus erythematosus, type I diabetes mellitus, and autoimmune liver disease. However, NK cells have both protective and pathogenic roles in autoimmunity depending on the NK cell subset, microenvironment, and disease type or stage. In this work, we review the current knowledge of the varied roles of NK cell subsets in systemic and organic-specific autoimmune diseases and their clinical potential as therapeutic targets.

The efficacy and adverse effects of the Uniblocker and left-side double-lumen tube for one-lung ventilation under the guidance of chest CT.

One-lung ventilation (OLV) is essential in numerous clinical procedures, in which the left-sided double-lumen tube (LDLT) is the most commonly used device. The application of bronchial blockers, including the Uniblocker or Arndt blocker, has increased in OLV. The present study aimed to compare the efficacy and adverse effects of the Uniblocker and LDLT for OLV under the guidance of chest CT. A total of 60 adult patients undergoing elective left-side thoracic surgery requiring OLV were included in the study. The patients were randomly assigned to the Uniblocker group (U group, n=30) or the LDLT group (D group, n=30). The time for initial tube placement, the number of optimal positions of the tube upon blind insertion, the number of attempts to adjust the tube to the optimal position, incidence of airway device displacement, injury to the bronchi and carina, the duration until lung collapse and the occurrence of sore throat and hoarseness over 24 h following surgery were recorded. The time for successful placement of the LDLT was 83.9±19.4 sec and that for the Uniblocker was 84.3±17.1 sec (P>0.05). The degree of lung collapse 1 min following opening of the pleura was greater in the D group than that in the U group (P<0.01) and the time required for the lung to completely collapse was shorter in the D group (3.3±0.5 min) than that in the U group (8.4±1.2 min; P<0.01). On the contrary, the incidence of injury to the bronchi and carina was lower in the U group (2/30 cases) than in the D group (10/30 cases; P=0.02); the incidence of sore throat was also lower in the U group (2/30 cases) compared with that in the D group (9/30 cases). The mean arterial pressure of patients immediately following intubation was lower in the U group (122.0±13.4 mmHg) than that in the D group (129.2±12.1 mmHg; P<0.05). The results of the present study indicated that the extraluminal use of the Uniblocker under guidance of chest CT is an efficient method with few adverse effects in left-side thoracic surgery. The study was registered at ClinicalTrials.gov on 16th December 2017 (no. NCT03392922).

Progranulin Regulates Inflammation and Tumor.

Progranulin (PGRN) mediates cell cycle progression and cell motility as a pleiotropic growth factor and acts as a universal regulator of cell growth, migration and transformation, cell cycle, wound healing, tumorigenesis, and cytotoxic drug resistance as a secreted glycoprotein. PGRN overexpression can induce the secretion of many inflammatory cytokines, such as IL-8, -6,-10, TNF-α. At the same time, this protein can promote tumor proliferation and the occurrence and development of many related diseases such as gastric cancer, breast cancer, cervical cancer, colorectal cancer, renal injury, neurodegeneration, neuroinflammatory, human atherosclerotic plaque, hepatocarcinoma, acute kidney injury, amyotrophic lateral sclerosis, Alzheimer's disease and Parkinson's disease. In short, PGRN plays a very critical role in injury repair and tumorigenesis, it provides a new direction for succeeding research and serves as a target for clinical diagnosis and treatment, thus warranting further investigation. Here, we discuss the potential therapeutic utility and the effect of PGRN on the relationship between inflammation and cancer.

pGreen-S: a clone vector bearing absence of enhanced green fluorescent protein for screening recombinants.

The bacterial cloning vector, pGreen-S, was constructed by inserting the enhanced green fluorescent protein (EGFP) gene at the XbaI restriction site of pUC18 plasmid. When expressed in Escherichia coli DH5alpha produced colonies that were an absinthe green color under daylight and strongly fluorescent green under longwave ultraviolet light. The pGreen-S vector was used to select for directional insert based on the loss of green fluorescence in recombinant colonies that was caused by the absence of EGFP. The EGFP reporter system differs from the conventional complementation of lacZ, making screening recombinants simpler, less expensive, and more effective.

A novel method of Uniblocker placement: extraluminal technique supported by trachea length measurement: A CONSORT-compliant article.

BACKGROUND: The use of bronchial blockers has been increased for one-lung ventilation; however, the placement of bronchial blockers is time consuming. The objective of this study was to compare the novel extraluminal technique of Uniblocker placement supported by trachea length measurement on computerized tomography images with conventional intraluminal Uniblocker placement method. METHODS: Seventy adult patients undergoing left side thoracic surgery were included in the study. All the patients were randomly assigned to one of two groups: conventional intraluminal intubation group (CV-IN group, n = 35) or extraluminal CT guided group (CT-EX group, n = 35). The primary endpoints were the optimal positions of Uniblocker and the injuries of bronchi and carina. The secondary outcomes included the time of Uniblocker placement, the adequacy of lung collapse, the incidences of Uniblocker displacement, sore throat, and hoarseness postoperative. RESULTS: In the CV-IN group, 19 of 35 Uniblockers went to the left main-stem bronchus on the initial blind insertion and 15 of 35 Uniblockers were considered as in optimal depth, whereas in the CT-EX group, 32 of 35 Uniblockers went to the left main-stem bronchus on the initial blind insertion and 31 of 35 Uniblockers were considered as in optimal depth (P < .01). The incidence of bronchi and carina injuries was obviously lower in the CT-EX group (occurred in 1 of 35 cases) than that in the CV-IN group (occurred in 8 of 35 cases) (P < .05). The time of Uniblocker placement took 145.4 s in the CV-IN group and 85.4 s in the CT-EX group (P < .01). The malpositions of Uniblocker, the degree of pulmonary collapse and the adverse events postoperative such as sore throat and hoarseness were not significantly different between the two groups (P > .05). CONCLUSION: The novel extraluminal technique of Uniblocker placement supported by trachea length measurement on computerized tomography images was proved to be more rapid, more accurate and less complications than conventional intraluminal Uniblocker placement method.