A Review of Strategies Associated with Surgical Decompression in Traumatic Spinal Cord Injury.

Traumatic spinal cord injury (TSCI) is frequent. Timely diagnosis and treatment have reduced the mortality, but the long-term recovery of neurologic functions remains ominous. After TSCI, tissue bleeding, edema, and adhesions lead to an increase in the intraspinal pressure, further causing the pathophysiologic processes of ischemia and hypoxia and eventually accelerating the cascade of secondary spinal cord injury. Timely surgery with appropriate decompression strategies can reduce that secondary injury. However, disagreement about the safety and effectiveness of decompression surgery and the timing of surgery still exists. The level and severity of spinal cord injury do have an impact on the timing of surgery; therefore, TSCI subpopulations may benefit from early surgery. Early surgery perhaps has little effect on recovery from complete TSCI but might be of benefit in patients with incomplete injury. Early decompression should be considered in patients with incomplete cervical TSCI. Patient age should not be used as an exclusion criterion for early surgery. The best time point for early surgery is although influenced by the shortest duration to thoroughly examine the patient's condition and stabilize the patient's state. After the patient's condition is fully evaluated, we can perform the surgical modality of emergency myelotomy and decompression. Therefore, a number of conditions should be considered, such as standardized decompression methods, indications and operation timing to ensure the effectiveness and safety of early surgical intervention, and promotion of the functional recovery of residual nerve tissue.

[The Value of MRI in Early Cervical Cancer Staging with Vaginal Filling of Ultrasound Gel].

OBJECTIVE: To investigate the value of MRI after ultrasonic gel vagina filling in the staging of early-stage cervical cancer. METHODS: A total of 158 patients with cervical cancer who underwent MRI examination after their cervical cancer diagnosis was confirmed by cervical biopsy were prospectively enrolled. Routine MRI examination was performed first, followed by another MRI examination after vaginal filling with ultrasound gel. Two physicians used a double-blind method to determine the staging of cervical cancer based on the MRI images before and after vaginal filling of ultrasound gel. Results of the postoperative pathology analysis were used as the golden standard. The positive predictive value and negative predictive value for stage Ⅱa cases of the two examinations of the same patient were compared, and the sensitivity, specificity and accuracy in identifying stage Ⅱa cervical cancer were compared. RESULTS: Two physicians used the double blind method to determine the staging of cervical cancer based on conventional MRI images, achieving moderate consistency ( κ=0.680). However, for the staging of cervical cancer with MRI images after vaginal filling of ultrasound gel, the two physicians achieved highly consistent results ( κ=0.932). Regarding identifying stage Ⅱa cervical cancer, the positive predictive value of conventional MRI was 66.67%, the negative predictive value was 76.74%, and the sensitivity, specificity and accuracy were 70.59%, 73.33% and 72.15%, respectively. The positive predictive value of MRI after vaginal filling of ultrasound gel was 90.91%, the negative predictive value was 91.3%, and the sensitivity, specificity and accuracy were 88.24%, 93.33% and 91.14%, respectively. The sensitivity, specificity and accuracy of the two methods were compared and the difference was statistical significant ( P<0.05). CONCLUSION: MRI examination after ultrasound gel vaginal filling has better diagnostic value for identifying stage Ⅱa cervical cancer. The method is simple and easy to do, and can be used in routine MRI examination of cervical cancer.

Overcoming Intrinsic H3K27me3 Imprinting Barriers Improves Post-implantation Development after Somatic Cell Nuclear Transfer.

Successful cloning by somatic cell nuclear transfer (SCNT) requires overcoming significant epigenetic barriers. Genomic imprinting is not generally regarded as such a barrier, although H3K27me3-dependent imprinting is differentially distributed in E6.5 epiblast and extraembryonic tissues. Here we report significant enhancement of SCNT efficiency by deriving somatic donor cells carrying simultaneous monoallelic deletion of four H3K27me3-imprinted genes from haploid mouse embryonic stem cells. Quadruple monoallelic deletion of Sfmbt2, Jade1, Gab1, and Smoc1 normalized H3K27me3-imprinted expression patterns and increased fibroblast cloning efficiency to 14% compared with a 0% birth rate from wild-type fibroblasts while preventing the placental and body overgrowth defects frequently observed in cloned animals. Sfmbt2 deletion was the most effective of the four individual gene deletions in improving SCNT. These results show that lack of H3K27me3 imprinting in somatic cells is an epigenetic barrier that impedes post-implantation development of SCNT embryos and can be overcome by monoallelic imprinting gene deletions in donor cells.

[Tracking Early Reperfused Myocardial Infarction using Cardiac MR].

OBJECTIVE: To determine segmental myocardial changes in cardiovascular magnetic resonance feature-tracking (CMR-FT) in the early phase of reperfused myocardial infarction in patients and rats. METHODS: Ten patients receiving percutaneous coronary interventions (2-10 d) and 10 rats with 60 min induced myocardial ischemia followed by reperfusions (48 h and 7 d) were investigated by MRI. The steady state free precession cine and late gadolinium enhancement (LGE) sequences were measured to evaluate the standard short axis of the whole heart after an injection of gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA, Magnevist, Bayer Health Care Pharmaceuticals) at a dose of 0.1 mmol/kg. The infarction sizes (all areas were expressed as a percentage of the whole myocardial tissues of left ventricle (LV), end-diastolic volume (EDV) and ejection fractions (EF) were calculated. The MRI cine images were analyzed using the myocardial feature tracking software CVI, estimating the peak value of radial strains (RS) and circumferential strains (CS) of the 16 AHA segments excluding apex cordis. The complete myocardial infarction (CMI) segments, partial myocardial infarction (PMI) segments and non-myocardial infarction (NMI) segments were identified and compared. RESULTS: Patients: The radial strain and circumferential strain of the CMI and PMI segments were smaller than the NMI segment (both P < 0.01). However, there was no significant difference between the CMI and the PMI segment (P>0.05). Rats: No significance differences were found in EF and EDV between the two time period 48 h and 7 d (both P>0.05). The radial strain and circumferential strain of the CMI and PMI segments were smaller than the NMI segment (all P < 0.01). But there was no significance difference between the CMI segment and the PMI segment (P>0.05). No significant changes in the global radial strain and the circumferential strain were found over time (both P>0.05). But the segmental radial strain and circumferential strain became larger over time (all P < 0.05). CONCLUSIONS: The systolic ability of myocardium decreases as a result of reperfusion injury in the early phase of reperfused myocardial infarction. But it can gradually recover over time with reperfusion.

Transplantation of adipose tissue-derived stem cells ameliorates autoimmune pathogenesis in MRL/lpr mice : Modulation of the balance between Th17 and Treg.

BACKGROUND: Mesenchymal stem cells (MSCs) are multipotent cells characterized by immunomodulatory properties and are therefore considered a promising tool for the treatment of autoimmune diseases. In this study, we aimed to investigate whether transplantation of adipose tissue-derived stem cells (ADSCs) affects the autoimmune pathogenesis in MRL/lpr mice. METHODS: Fifteen 12-week-old MRL/lpr mice were randomly divided into three groups: ADSC, cyclophosphamide (CTX), and control groups, with five mice in each group. ADSC and control groups were injected with 1â€¯× 106 ADSCs or PBS, respectively, via the tail vein, once a week for 8 weeks. The CTX group was injected with CTX at a dose of 15 mg/kg body weight, once a week for 2 weeks, and this was repeated after 2 weeks rest. Proteinuria, anti-double-stranded DNA (anti-dsDNA) antibody, and serum creatinine levels were then measured. The populations of Th17 and Treg cells in the spleen were detected by flow cytometry. All statistical analyses were performed using least square difference. RESULTS: Eight weeks after treatment, the 24 h proteinuria, anti-dsDNA antibody levels, and serum creatinine were decreased significantly with transplantation of mouse ADSCs. ADSCs markedly reduced the number of TH17 cells, increased Treg cells, and improved renal pathology. CONCLUSION: Our results indicate that transplantation of ADSCs could significantly inhibit autoimmune progression in MRL/lpr mice and the efficacy of ADSCs was comparable to that of CTX.

Integrated mapping of pharmacokinetics and pharmacodynamics in a patient-derived xenograft model of glioblastoma.

Therapeutic options for the treatment of glioblastoma remain inadequate despite concerted research efforts in drug development. Therapeutic failure can result from poor permeability of the blood-brain barrier, heterogeneous drug distribution, and development of resistance. Elucidation of relationships among such parameters could enable the development of predictive models of drug response in patients and inform drug development. Complementary analyses were applied to a glioblastoma patient-derived xenograft model in order to quantitatively map distribution and resulting cellular response to the EGFR inhibitor erlotinib. Mass spectrometry images of erlotinib were registered to histology and magnetic resonance images in order to correlate drug distribution with tumor characteristics. Phosphoproteomics and immunohistochemistry were used to assess protein signaling in response to drug, and integrated with transcriptional response using mRNA sequencing. This comprehensive dataset provides simultaneous insight into pharmacokinetics and pharmacodynamics and indicates that erlotinib delivery to intracranial tumors is insufficient to inhibit EGFR tyrosine kinase signaling.

Monocrystalline platinum-nickel branched nanocages with enhanced catalytic performance towards the hydrogen evolution reaction.

Single crystalline noble metal nanocages are the most promising candidates for heterogeneous catalysis due to their large specific surface area, well-defined structure and enhanced structural stability. Herein, based on the observation of an unexpected phenomenon that the alloying of Pt and transition metals by co-reduction is more preferential than the formation of pure Pt NCs, we propose a feasible one-pot strategy to synthesize a uniformly epitaxial core-shell Pt-Ni structure with a Ni-rich alloy as the core and a Pt-rich alloy as the shell. The as-prepared Pt-Ni core-shell structures are subsequently etched into monocrystalline Pt-Ni branched nanocages with the wall thickness being 2.8 nm. This unique structure exhibits excellent catalytic performance and stability for the hydrogen evolution reaction (HER) in alkaline solution which is of great significance for the energy-intensive water-alkali and chlor-alkali industry.

Simultaneous two-color stimulated Raman scattering microscopy by adding a fiber amplifier to a 2 ps OPO-based SRS microscope.

Stimulated Raman scattering (SRS) microscopy is a label-free chemical imaging technique. Two-color imaging is often necessary to determine the distribution of chemical species in SRS microscopy. Current multi-color SRS imaging methods involve complicated instrumentation or longer data acquisition time or are limited to transmission imaging. In this Letter, we show that by adding a simple fiber amplifier to a 2 ps laser source and optical-parametric-oscillator-based SRS setup, one can achieve simultaneous two-color or frequency modulation SRS microscopy. The fiber amplifier can generate a wavelength tunable laser of ±10 nm around the Stokes laser wavelength at 1031 nm with average power greater than 200 mW. In vivo and ex vivo lipid-protein imaging of mouse brain and skin is demonstrated. To further demonstrate the potential of this technique in high-speed in vivo imaging, white blood cells in a blood stream are imaged in a live mouse.

Monitoring peripheral nerve degeneration in ALS by label-free stimulated Raman scattering imaging.

The study of amyotrophic lateral sclerosis (ALS) and potential interventions would be facilitated if motor axon degeneration could be more readily visualized. Here we demonstrate that stimulated Raman scattering (SRS) microscopy could be used to sensitively monitor peripheral nerve degeneration in ALS mouse models and ALS autopsy materials. Three-dimensional imaging of pre-symptomatic SOD1 mouse models and data processing by a correlation-based algorithm revealed that significant degeneration of peripheral nerves could be detected coincidentally with the earliest detectable signs of muscle denervation and preceded physiologically measurable motor function decline. We also found that peripheral degeneration was an early event in FUS as well as C9ORF72 repeat expansion models of ALS, and that serial imaging allowed long-term observation of disease progression and drug effects in living animals. Our study demonstrates that SRS imaging is a sensitive and quantitative means of measuring disease progression, greatly facilitating future studies of disease mechanisms and candidate therapeutics.

Label-Free Neurosurgical Pathology with Stimulated Raman Imaging.

The goal of brain tumor surgery is to maximize tumor removal without injuring critical brain structures. Achieving this goal is challenging as it can be difficult to distinguish tumor from nontumor tissue. While standard histopathology provides information that could assist tumor delineation, it cannot be performed iteratively during surgery as freezing, sectioning, and staining of the tissue require too much time. Stimulated Raman scattering (SRS) microscopy is a powerful label-free chemical imaging technology that enables rapid mapping of lipids and proteins within a fresh specimen. This information can be rendered into pathology-like images. Although this approach has been used to assess the density of glioma cells in murine orthotopic xenografts models and human brain tumors, tissue heterogeneity in clinical brain tumors has not yet been fully evaluated with SRS imaging. Here we profile 41 specimens resected from 12 patients with a range of brain tumors. By evaluating large-scale stimulated Raman imaging data and correlating this data with current clinical gold standard of histopathology for 4,422 fields of view, we capture many essential diagnostic hallmarks for glioma classification. Notably, in fresh tumor samples, we observe additional features, not seen by conventional methods, including extensive lipid droplets within glioma cells, collagen deposition in gliosarcoma, and irregularity and disruption of myelinated fibers in areas infiltrated by oligodendroglioma cells. The data are freely available in a public resource to foster diagnostic training and to permit additional interrogation. Our work establishes the methodology and provides a significant collection of reference images for label-free neurosurgical pathology. Cancer Res; 76(12); 3451-62. ©2016 AACR.

Optical technologies for intraoperative neurosurgical guidance.

Biomedical optics is a broadly interdisciplinary field at the interface of optical engineering, biophysics, computer science, medicine, biology, and chemistry, helping us understand light-tissue interactions to create applications with diagnostic and therapeutic value in medicine. Implementation of biomedical optics tools and principles has had a notable scientific and clinical resurgence in recent years in the neurosurgical community. This is in great part due to work in fluorescence-guided surgery of brain tumors leading to reports of significant improvement in maximizing the rates of gross-total resection. Multiple additional optical technologies have been implemented clinically, including diffuse reflectance spectroscopy and imaging, optical coherence tomography, Raman spectroscopy and imaging, and advanced quantitative methods, including quantitative fluorescence and lifetime imaging. Here we present a clinically relevant and technologically informed overview and discussion of some of the major clinical implementations of optical technologies as intraoperative guidance tools in neurosurgery.

[Effects of ischemic preconditioning on the serum testosterone level and spermatogenic cell apoptosis in rabbits with testicular ischemia-reperfusion].

OBJECTIVE: To explore the effects of ischemic preconditioning on the level of serum testosterone (T) and apoptosis of spermatogenic cells in rabbits with testicular ischemia-reperfusion injury induced by testicular torsion. METHODS: A total of 15 New Zealand male rabbits were randomly divided into groups A (control), B (ischemia-reperfusion), and C (ischemic preconditioning). The animals of group A were subjected to exposure of the right spermatic cord without ischemia, those of group B received 60-minute non-invasive occlusion of the right spermatic cord followed by 3 days of reperfusion, and those of group C underwent 5-minute occlusion plus 5-minute reperfusion of the right spermatic cord followed by the same procedure as that for group B. Then the rabbits were narcotized with 3% barbital sodium, the whole blood collected for examination of the serum T content and the testis tissues obtained from both the ischemic and healthy sides for HE and TUNEL staining. RESULTS: After operation, the body weight was significantly increased as compared with the baseline in groups A (ï¼»2.65±0.07ï¼½ vs ï¼»2.45±0.07ï¼½ kg, P<0.05) and C (ï¼»3.03±0.11ï¼½ vs ï¼»2.92±0.07ï¼½ kg, P<0.05), but not in group B (ï¼»3.05±0.07ï¼½ vs ï¼»3.05±0.07ï¼½ kg, P>0.05). The serum T level showed no statistically significant difference in group A before and after operation (ï¼»139.59±9.39ï¼½ vs ï¼»140.19±9.47ï¼½ ng/L, P>0.05), but was remarkably lower after operation than the baseline in groups B ï¼»148.06±3.31ï¼½ vs ï¼»74.12±4.00ï¼½ ng/L, P<0.01) and C (ï¼»133.75±6.48ï¼½ vsï¼»94.76±3.13ï¼½ ng/L, P<0.01) as well as than the postoperative index in group A (P<0.01). In comparison with group A and the healthy side of group B, the testis tissue of the ischemic side in group B exhibited structural damage of most of the seminiferous tubules with disappearance of spermatogenic cell structures, apoptosis of spermatogenic cells, and exudation of light-eosin edema fluid in the mesenchyme and lumen, with a markedly increased apoptosis index (P<0.01) and a significantly decreased Johsen's score (P<0.01). Compared with ischemic side of group B, The testis tissue of the ischemic side in group C was restored to normal as compared with that in group B, with a dramatically decreased apoptosis index (P<0.01) and a remarkably increased Johnsen's score (P<0.01). CONCLUSIONS: Ischemic preconditioning can raise the decreased serum T level and reduce the apoptosis of spermatogenic cells in rabbits with testicular ischemia-reperfusion injury, which could be applied as a potential option for the clinical treatment of testicular ischemia-reperfusion injury.

Label-free DNA imaging in vivo with stimulated Raman scattering microscopy.

Label-free DNA imaging is highly desirable in biology and medicine to perform live imaging without affecting cell function and to obtain instant histological tissue examination during surgical procedures. Here we show a label-free DNA imaging method with stimulated Raman scattering (SRS) microscopy for visualization of the cell nuclei in live animals and intact fresh human tissues with subcellular resolution. Relying on the distinct Raman spectral features of the carbon-hydrogen bonds in DNA, the distribution of DNA is retrieved from the strong background of proteins and lipids by linear decomposition of SRS images at three optimally selected Raman shifts. Based on changes on DNA condensation in the nucleus, we were able to capture chromosome dynamics during cell division both in vitro and in vivo. We tracked mouse skin cell proliferation, induced by drug treatment, through in vivo counting of the mitotic rate. Furthermore, we demonstrated a label-free histology method for human skin cancer diagnosis that provides comparable results to other conventional tissue staining methods such as H&E. Our approach exhibits higher sensitivity than SRS imaging of DNA in the fingerprint spectral region. Compared with spontaneous Raman imaging of DNA, our approach is three orders of magnitude faster, allowing both chromatin dynamic studies and label-free optical histology in real time.

Identification of a functional nuclear localization signal mediating nuclear import of the zinc finger transcription factor ZNF24.

ZNF24 is a member of the SCAN domain family of Krüppel-like zinc finger (ZF) transcription factors, which plays a critical role in cell proliferation and differentiation. However, how ZNF24 enters the nucleus in order to exert its function remains unclear since its nuclear localization signal(s) (NLS) has not been identified. Here, we generated a series of GFP-tagged deletion and point mutants and assessed their subcellular localization. Our results delimit the NLS to ZF1-2. Deletion of ZF1-2 caused cytoplasmic accumulation of ZNF24. Fusion of the ZF1-2 to green fluorescent protein (GFP) targeted GFP to the nucleus, demonstrating that the ZF1-2 is both necessary and sufficient for nuclear localization. ZNF24 containing histidine to leucine mutations that disrupt the structure of ZF1 or/and ZF2 retains appropriate nuclear localization, indicating that neither the tertiary structure of the zinc fingers nor specific DNA binding are necessary for nuclear localization. K286A and R290A mutation led to partial cytoplasmic accumulation. Co-immunoprecipitation demonstrated that ZNF24 interacted with importin-ß and this interaction required the ZF motifs. The ß-Catenin (CTNNB1) luciferase assays showed that the ZNF24 mutants defective in nuclear localization could not promote CTNNB1 promoter activation as the wild-type ZNF24 did. Taken together, these results suggest that consecutive ZF1-2 is critical for the regulation of ZNF24 nuclear localization and its transactivation function.

[Effect of acupuncture stimulation of Zusanli (ST 36) on cerebral regional homogeneity in volunteer subjects with different constitutions: a resting-state fMRI study].

OBJECTIVE: To observe the synchronism difference of brain region activities in response to acupuncture stimulation of Zusanli (ST 36) in healthy volunteer subjects with different acupuncture analgesia sensitivity, so as to study the central factors influencing acupuncture intervention outcomes. METHODS: Forty-five healthy volunteer subjects with different constitutions (different sensitivities in response to needling stimulation) were divided into insensitive group, normal group and sensitive group (n = 15). The pressure pain threshold (PPT) of the Zusanli (ST 36) region before and after acupuncture stimulation of ST36 was assessed using visual analog scale (VAS). Two weeks later after acupuncture stimulation of ST 36, resting-state fMRI images were acquired by using a nuclear magnetic resonance imaging system and analyzed by using DPARSFV 2.1 software package, software SPM 8 and REST 1.7. The cerebral regional homogeneity (ReHo) of the subjects was then calculated by Resting-State fMRI Data Analysis Toolkit (REST). RESULTS: Compared with pre-acupuncture, PPT levels of the normal and sensitive groups were significantly increased after acupuncture of ST 36 (P < 0.05), and that of the insensitive group had no significant change (P > 0.05). Following acupuncture stimulation of ST 36, the insensitive group only showed a significant decreased ReHo in the left fusiform gyrus, left inferior temporal gyrus, bilateral postcentral gyrus, and left anterior central gyrus. In the normal group, a significantly increased ReHo was found in left brainstem, the right cerebellum posterior lobe, right parahippocampa gyrus, right fusiform gyrus, left angular gyrus, temporal lobe and the left frontal lobe; and a significantly decreased ReHo in the occipital lobes and the right superior temporal gyrus after acupuncture stimulation of ST 36. In the sensitive group, a markedly increased ReHo was found in the left brainstem, bilateral cerebellum posterior lobes, left inferior temporal gyrus, basal ganglia, the left insular lobe, anterior cingutate, frontal lobe, inferior parietal lobule, and the right supplementary motor area, and an obviously decreased ReHo found in the bilateral occipital lobes, fusiform gyrus, posterior central gyrus, the right posterior cingutate, the left temporal lobe and the left paracentral lobule, etc. after acupuncture of ST 36. CONCLUSION: Constitution-associated needling sensation may be an important influential factor for acupuncture analgesia in normal subjects. The change of ReHo in different cerebral areas is probably responsible for the difference of acupuncture analgesia in different constitution people.

[An experimental study of contrast enhanced ultrasound diagnosis of cerebral intraparenchymal hemorrhage in dogs].

OBJECTIVE: To explore the role of contrast enhanced ultrasound (CEUS) in diagnosing experimental cerebral intraparenchymal hemorrhage (IH) in dogs. METHODS: A self-control study was conducted. An IH model was reproduced by puncturing middle cerebral artery (MCA) in 12 dogs. Two-dimensional ultrasound and CEUS were conducted immediately, 30 minutes, and 1 hour after modeling, respectively, to observe the lesion echo and bleeding area. CT scans were also conducted at 1 hour after modeling, then the lesion size in CT scan was compared with that of CEUS. RESULTS: In 12 dogs IH model was reproduced successfully, and unilateral hematomas were confirmed by CT and pathological examination. Two-dimensional ultrasound of IH showed irregular high-echo area, with unclear boundary, but it was not able to show active bleeding. CEUS demonstrated active bleeding by outflow and pooling of contrast agent with obvious enhancement. CEUS of the hematoma showed perfusion deficit, with a clear boundary. The size of bleeding lesions (cm) continued to increase at 30 minutes and 1 hour after modeling (1.47±0.40, 1.76±0.45 by CEUS measuring), and demonstrated statistically significant difference comparing with the measurement of IH immediately after modeling (1.03±0.24, both P<0.01), while there was no statistically significant difference between the 30-minute and 1-hour measurements (P>0.05). Compared with the measurements between CEUS and CT at 1 hour, there was no statistically significant difference in size of bleeding lesions (1.76±0.45 vs. 1.79±0.47, P>0.05). CONCLUSION: CEUS can help determine the extent and size of IH, and the process of hematoma formation when dynamically monitoring.

Quantitative chemical imaging with multiplex stimulated Raman scattering microscopy.

Stimulated Raman scattering (SRS) microscopy is a newly developed label-free chemical imaging technique that overcomes the speed limitation of confocal Raman microscopy while avoiding the nonresonant background problem of coherent anti-Stokes Raman scattering (CARS) microscopy. Previous demonstrations have been limited to single Raman band measurements. We present a novel modulation multiplexing approach that allows real-time detection of multiple species using the fast Fourier transform. We demonstrate the quantitative determination of chemical concentrations in a ternary mixture. Furthermore, two imaging applications are pursued: (1) quantitative determination of oil content as well as pigment and protein concentration in microalgae cultures; and (2) 3D high-resolution imaging of blood, lipids, and protein distribution in ex vivo mouse skin tissue. We believe that quantitative multiplex SRS uniquely combines the advantage of fast label-free imaging with the fingerprinting capability of Raman spectroscopy and enables numerous applications in lipid biology as well as biomedical imaging.

Multicolor stimulated Raman scattering (SRS) microscopy.

Stimulated Raman scattering (SRS) microscopy has opened up a wide range of biochemical imaging applications by probing a particular Raman-active molecule vibrational mode in the specimen. However, the original implementation with picosecond pulse excitation can only realize rapid chemical mapping with a single Raman band. Here we present a novel SRS microscopic technique using a grating-based pulse shaper for excitation and a grating-based spectrograph for detection to achieve simultaneous multicolor SRS imaging with high sensitivity and high acquisition speeds. In particular, we used linear combination of the measured CH2 and CH3 stretching signals to map the distributions of protein and lipid contents simultaneously.

Haemostatic agents of the gelatin matrix for a large liver wound by percutaneous injection without pressure under the guidance of contrast-enhanced ultrasound.

BACKGROUND: The non-operation treatment of intra-abdominal trauma guided contrast enhanced ultrasound (CEUS) is one of the hottest research topic. Gelatin/thrombin/calcium (GTC) was developed as a novel haemostatic agent for non-operable intra-abdominal trauma. We hypothesized that GTC can achieve haemostasis (without the use of pressure) within a short time in a large wound model by percutaneous injection under CEUS guidance. METHODS: Forty Wister rats received large liver injuries by haemostatic clamp and were randomly divided into four groups, according to the haemostatic agent used. These included normal saline (NS) group A, lyophilising thrombin powder (LTP) group B, GTC group C, and absorbable α-cyanoacrylate (ACNA) group D. Each injury site was treated with one of the above materials and total bleeding time was recorded. All liver wounds were evaluated using CEUS at three periods: pre-injury, injury and post-treatment. The liver wounds were also evaluated by histology 3, 6, and 9 days after injury and the extents of abdominal adhesions were recorded. RESULTS: The sensitivity of CEUS (100%) in detecting blunt traumatic liver lesions was significantly higher than conventional ultrasound (42.5%). Bleeding times at the injury site in the GTC group C ((129.3 ± 14.0) seconds) and ACNA group D ((5.2 ± 1.0) seconds) were significantly shorter than those in the NS group A ((369.5 ± 48.8) seconds, P < 0.01) and LTP group B ((324.7 ± 52.22) seconds, P < 0.01). The LTP group B showed no significant difference compared with the NS group A. Gross examination of liver tissue revealed that there were fewer intra-abdominal adhesions in the GTC group C (10%) than in the ACNA group D (100%). Histopathologic examination showed that GTC was completely absorbed after nine days. CONCLUSIONS: GTC, delivered by percutaneous injection under CEUS, may achieve haemostasis (without the use of pressure) within a short time in a large wound model. GTC is absorbable and may prevent intra-abdominal adhesions. Therefore, it may be the optimal choice for first aid treatment of large abdominal wounds in the setting of blunt trauma.

Characterization of blood components separated from donated whole blood after an overnight holding at room temperature with the buffy coat method.

BACKGROUND: With buffy coat (BC) processing of whole blood (WB) donations, increase in WB storage time to facilitate overnight holding before the separation of blood components would be a logistically attractive development. This study undertakes a comparative in vitro characterization of blood components prepared from WB samples that were either processed within 8 hours or stored overnight at room temperature before processing by the BC method. STUDY DESIGN AND METHODS: The WB units (400 mL) collected were either processed within 8 hours (fresh blood) or stored overnight (overnight blood) at room temperature. WB units were separated into individual-component red blood cells (RBCs), BC, and plasma. The in vitro quality of these blood components (RBCs, pooled platelet concentrates [PCs], and plasma) was analyzed during storage. RESULTS: Levels of 2,3-diphosphoglycerate (2,3-DPG) were found to be significantly lower immediately after processing, compared with the fresh WB samples, in RBCs that had been separated from an overnight-hold sample. However, this difference was not apparent after 14 days of storage. In pooled PCs, measurements for glucose, lactate, PO(2), PCO(2), extent of shape change, and hypotonic shock response were similar. The platelet yield in PCs prepared from an overnight-hold WB sample was significantly higher, while CD62P expression and annexin V binding were lower (p < 0.05). For frozen plasma (FP), no significant differences were observed for the coagulation factors (F)II, FVII, FV, F IX, FX, and FXI; fibrinogen; and von Willebrand factor content between the 8- and 24-hour FP. The FVIII was the component that was most sensitive to the prolongation of production time and it only had 80% of the activity of the 8-hour FP. CONCLUSION: These data suggest that blood components (RBCs, pooled PCs, and FP) separated from WB that has been stored overnight at room temperature by the BC method are of acceptable quality.