Differentiation of Human GBM From Non-GBM Brain Tissue With Polarization Imaging Technique.

As for optical techniques, it is difficult for the 5-aminolevulinic (5-ALA) fluorescence guidance technique to completely detect glioma due to residual cells in the blind area and the dead angle of vision under microscopy. The purpose of this research is to characterize different microstructural information and optical properties of formalin-soaked unstained glioblastoma (GBM) and non-GBM tissue with the polarization imaging technique (PIT), and provide a novel method to detect GBM during surgery. In this paper, a 3×3 Mueller matrix polarization experimental system in backscattering mode was built to detect the GBM and non-GBM tissue bulk. The Mueller matrix decomposition and transformation parameters of GBM and non-GBM tissue were calculated and analyzed, and showed that parameters (1- Δ ) and t are good indicators for distinguishing GBM from non-GBM tissues. Furthermore, the central moment coefficients (CMCs) of the frequency distribution histogram (FDH) were also calculated and used to distinguish the cancerous tissues. The results of the experiments confirmed the feasibility of PIT applied in the clinic to detect glioma, laying the foundation for the subsequent non-invasive, non-staining glioma detection.

Identification of two novel mutations in three Chinese families with Kallmann syndrome using whole exome sequencing.

Kallmann syndrome (KS) is a rare developmental disorder that manifests as congenital hypogonadotropic hypogonadism with anosmia. More than 19 genes have been found to be associated with KS. However, approximately 70% of the causes of KS remain unclear. Here, we studied seven KS patients, from three families, who had delayed puberty and olfactory bulb dysplasia. However, the families of these patients showed a range of other unique clinical features, including hearing loss, anosmia (to varying degrees) and unilateral renal agenesis. We performed whole exome sequencing and copy number variation (CNV) sequencing on samples acquired from these patients. We identified two novel mutations (c.844delC in ANOS1, c.475C>T in SOX10) and a novel trigenic pattern, PROKR2/CHD7/FEZF1 (c.337T>C in PROKR2, c.748C>G in FEZF1, c.8773G>A in CHD7). The c.844delC mutation in the ANOS1 gene was predicted to generate a truncated form of the anosmin-1 protein. SIFT and PolyPhen-2 predicted that the c.475C>T mutation in SOX10 had a damaging effect. The PROKR2 mutation (c.337T>C) was previously reported as harmful. No pathogenic copy number alterations were detected. Our study expands the genotypic and phenotypic spectrum of KS, a disease that shows considerable clinical and genetic heterogeneity. The application of whole exome sequencing could facilitate our understanding of the pathogenesis of KS.

A novel Laccase gene from Litopenaeus vannamei is involved in the immune responses to pathogen infection and oxidative stress.

Laccases (Lacs) are copper-containing oxidase enzymes that are found in various plants, fungi, and microorganisms. For invertebrates, particularly insects and crustaceans, Lacs have been shown to be involved in immune responses. In shrimp, a Lac gene has been cloned and functionally characterized, which revealed that it is involved in shrimp anti-pathogen infection. In the present study, a novel Lac gene (LvLac2) was cloned from Litopenaeus vannamei. Real-time RT-PCR analysis showed that LvLac2 is induced by white spot syndrome virus (WSSV)- or Vibrio alginolyticus infection. In addition, the downregulated expression of LvLac2 decreased the cumulative mortality of WSSV- or V. alginolyticus infected shrimps. Moreover, LvLac2 is also induced by oxidative stress. Knocking down the expression of LvLac2 decreased the severity of hepatopancreatic injury caused by oxidative stress, as well as reduced the cumulative shrimp mortality during oxidative stress. Furthermore, gene reporter assays showed that the expression of LvLac2 is regulated by NF-E2-related factor 2, which is the key transcription factor of the oxidative stress response signaling pathway. Our study revealed that LvLac2 not only participates in immune responses against infections in L. vannamei but is also involved in oxidative stress responses.

Identification and functional characterization of a C-type lectin gene from Litopenaeus vannamei that is associated with ER-stress response.

C-type lectins (CTLs), which bind carbohydrates in a Ca2+-dependent manner, are involved in many cellular activities, especially immunity. CTLs play important roles in both the antibacterial and the antiviral immune response and are also associated with autoimmunity. Several CTLs have been investigated in crustaceans, primarily with respect to their function in the immune response. In this study, we cloned a novel CTL gene (LvCTLU) from Litopenaeus vannamei. LvCTLU is involved in microbe agglutination and phagocytosis. Downregulating LvCTLU increased the cumulative mortality of L. vannamei after Vibrio parahemolyticus infection. Similar to other reported CTLs, LvCTLU also had antiviral properties. Downregulation of LvCTLU also increased the cumulative mortality of L. vannamei after infection with white spot syndrome virus. More importantly, LvCTLU expression was induced by the unfolded protein response (UPR), which is the key pathway in the endoplasmic reticulum (ER)-stress response of eukaryotic organism. Our results suggested that this protein might be involved in the shrimp ER-stress response. Reporter gene assay indicated that LvCTLU was regulated by X-box-binding protein 1, which is the key transcription factor in the UPR. Our study thus revealed that LvCTLU plays vital roles in both the anti-pathogen immune response and the ER-stress response.

Functional characterization of a matrix metalloproteinase 2 gene in Litopenaeus vannamei.

Matrix metalloproteinases (MMPs) contribute to both normal and pathological tissue remodeling. They act as regulatory molecules by working in enzyme cascades as well as processing matrix proteins, cytokines, growth factors and adhesion molecules to generate fragments with biological effects. So MMPs could play distrinct roles in the process of pathogen infection. In present study, we cloned a MMP-2 (LvMMP-2) gene from Litopenaeus vannamei. LvMMP-2, highly expressed in epidermis, located to endoplasmic reticulum in S2 cells. Results of real-time RT-PCR assay showed that LvMMP-2 was induced in shrimp hemocytes upon unfolded protein response or oxidative stress, but not via heat shock treatment. It is proved that the promoter activity of LvMMP-2 was enhanced by NF-E2-related factor 2 and AP-1 factor c-Jun. Further research showed that down-regulated LvMMP-2 contributing to oxidative stress injury, could reduce the cumulative mortality of shrimps under oxidative stress. Besides, our study also indicated that LvMMP-2 was accelerated by lipopolysaccharides injection. LvMMP-2 in S2 could increase the promoter activity of several antimicrobial peptide genes, and knocked-down expression of LvMMP-2 depressed the expression of penaeidin2 and ß-Defensin. Moreover, we showed that down-regulated LvMMP-2 suppressed the cumulative mortality of shrimp infected with white spot syndrome virus (WSSV) or with Vibrio alginolyticus. Collecting results suggested that LvMMP-2 involves in shrimp innate immune response, and also contributes to tissue injury caused by WSSV infection.

Functional characterization of an ER-stress responding Crustin gene in Litopenaeus vannamei.

Shrimp in culture ponds are challenged by various pathogens as well as harsh water environment. The innate immune system and environmental stress response system of shrimp paly an important role in shrimp survival and growth. For remission the endoplasmic reticulum (ER)-stress caused by environmental stress, unfolded protein response (UPR) may reduce the synthesis of most proteins, including great mass of immune factors, which could weaken the immune function of shrimp. Therefore, how cells keep appropriate amount of immune factor synthesis under such a situation is critical important for shrimp health and growth. In this study, we cloned a new Crustin gene (LvCruU) from Litopenaeus vannamei. We showed that LvCruU has antibacterial activity, and reducing its expression would increase the cumulative mortality of L. vannamei upon the Vibrio parahemolyticus infection. In addition, we found that promoter activity of LvCruU was enhanced not only by the deformed epidermal autoregulatory factor-1 (Deaf1), but also by activating transcription factor 3 (LvATF3) of shrimp UPR. Real-time RT-PCR showed that LvCruU and LvATF3 both were induced upon UPR activation. And moreover, in Thapsigargin plus dsLvCruU injection test, we showed that down-regulation of LvCruU increased the cumulative mortality of V. parahemolyticus-infected shrimp under ER-stress. These results suggest that LvCruU work as a downstream effector of UPR, and contribute to antimicrobic immune response upon ER-stress in L. vannamei.

Imbalance of T-helper 1/T-helper 2 cytokines and impaired glucose tolerance among patient with acute coronary syndrome.

PURPOSE: The balance between T helper (Th) cells Th1- and Th2-related cytokines plays a key role in the clinical process of acute coronary syndrome (ACS) and type 2 diabetes mellitus (T2DM) or impaired glucose tolerance (IGT). The objective of this study was to assess the status of Th1/Th2 cytokines in patients with ACS and T2D or IGT. METHODS: A total of 201 ACS patients were enrolled in the study. All ACS patients were divided into three groups: Group I-patients with normal glucose tolerance (NGT), Group II-patients with IGT and Group III-patients with T2D. We measured circulating Th1/Th2-type cytokines (interleukin [IL]-4, IL-13, interferon-gamma [IFN-γ], and tumor-necrosis factor-alpha [TNF-α]) using enzyme-linked immunosorbent assay and calculated the ratio of Th1/Th2. RESULTS: Significant elevations in serum levels of IL-4, IL-13, IFN-γ, and TNF-α were found in ACS-T2D and ACS-IGT groups compared to that in both ACS-NGT group and healthy individuals. Higher serum levels of IL-4, IL-13, and TNF-α were found in ACS-NGT group than that in the control group. Furthermore, IL-4 and IFN-γ concentrations were significantly higher in ACS-T2D patients than in ACS-IGT patients. IFN-γ/IL-4, IFN-γ/IL-13, and TNF-α/IL-4 ratios as markers of Th1/Th2 ratio were significantly higher for the ACS-T2D group and ACS-IGT group as compared to that in the ACS-NGT group and control group (P < 0.05). CONCLUSION: Shifts in the balance of Th1/Th2 toward a predominance of Th1 may represent more severe inflammatory status in ACS patients with type T2D or IGT.

Efficacy of internal fixation with mini plate and internal fixation with hollow screw for Regan-Morrey type II and III ulna coronoid fractures.

BACKGROUND: Ulna coronoid fracture is a complicated injury and occurred in the coronal plane. Undeniably, there is no universally accepted approach for treating ulna coronoid fractures. Therefore, this study aimed at exploring the efficacy of different surgical treatments for Regan-Morrey type II and III ulna coronoid fractures. METHODS: A total of 164 patients with ulna coronoid fractures were admitted and treated in department of orthopedics at Yiwu Central Hospital, the Affiliated Yiwu Hospital of Wenzhou Medical University for retrospective analysis. The baseline features (age, gender, time from injury to surgery and so on) before the surgery and different conditions during the surgery were compared. Following that, the Visual Analogue Scale (VAS) pain score was employed to evaluate the severity of preoperative and postoperative pain experienced by the patients in each group. Afterwards, Broberg and Morrey elbow score was used to evaluate elbow joint function and surgical effect of the patients. Lastly, the postoperative recovery and complications were compared. RESULTS: It was firstly observed that internal fixation with mini plate and hollow screw compelled to lower average operation time and blood loss than Kirschner wire and steel wire suture. Next, the severity of postoperative pain was lessened in comparison with preoperative pain. Afterwards, mini plate and hollow screw improved elbow joint function more notable than Kirschner wire and steel wire suture, and Kirschner wire and steel wire suture resulted in higher incidence of complications and worse postoperative recovery. CONCLUSION: Collectively, this study clarified that for the treatment of Regan-Morrey type II and III ulna coronoid fractures, internal fixation with mini plate and hollow screw has an overall superior surgical effect than internal fixation with Kirschner wire and steel wire suture.

Litopenaeus vannamei activating transcription factor 6 alpha gene involvement in ER-stress response and white spot symptom virus infection.

A previous study found that inositol-requiring enzyme-1-X-box binding protein 1 (IRE1-XBP1) pathway and the protein kinase RNA (PKR)-like ER kinase-eIF2α (PERK-eIF2α) pathway of shrimp play roles in the unfolded protein response (UPR). And they also be proved that was involved in white spot symptom virus (WSSV) infection. Yet the functions of the third branch in shrimp UPR are still unclear. In this study, we showed that upon UPR activation, activating transcription factor 6 alpha (LvATF6α) of Litopenaeus vannamei was cleaved and transferred from the cytoplasm to the nucleus in 293T cells, indicating that the ATF6 pathway in shrimp is also a branch of UPR. Furthermore, LvATF6α could reduce the apoptosis rate of Drosophila Schneider 2 (S2) cells treated with actinomycin, and knock-down expression of LvATF6α increased the apoptosis rate of shrimp hemocytes. In vivo testing revealed that the short from LvATF6α (LvATF6α-s) was obviously increased after UPR activation or WSSV infection, indicating that the ATF6 pathway was activated in L. vannamei gills under such circumstances. Moreover, knock-down expression of LvATF6α could reduce the cumulative mortality and WSSV copy number in WSSV-infected shrimp. Further study revealed that WSSV may profit from shrimp ATF6 pathway activation in two aspects. First, LvATF6α-s significantly upregulated the expression of the WSSV genes (wsv023, wsv045, wsv083, wsv129, wsv222, wsv249, and wsv343). Second, LvATF6α-s inhibited apoptosis by negatively regulating the apoptosis signal-regulating kinase 1 - (c-Jun N-terminal kinase) pathway. All of these evidences suggested that the ATF6 pathway is a member of the L. vannamei UPR, and it is also engaged in WSSV infection.

Functional characterization of a reactive oxygen species modulator 1 gene in Litopenaeus vannamei.

Reactive oxygen species (ROS) imparts a dual effect on multicellular organisms, wherein high levels are usually harmful, and low levels could facilitate in combating pathogenic microorganisms; therefore, the regulation of ROS production is critical. Previous studies have suggested that ROS contributes to resistance to the white spot syndrome virus (WSSV) or Vibrio alginolyticus in Litopenaeus vannamei. However, the regulation of ROS metabolism in L. vannamei remains elusive. In the present study, we proved that the overexpression of L. vannamei reactive oxygen species modulator 1 (LvROMO1) increases ROS production in Drosophila Schneider 2 (S2) cells. Real-time RT-PCR analysis indicated that LvROMO1 is induced by WSSV or V. alginolyticus infection and ß-glucan or microcystin (MC-LR) injection. Further investigation showed that LvROMO1 responding to MC-LR, thereby inducing hemocytes to undergo apoptosis, and ultimately resulting in hepatopancreatic damage. And LvROMO1 downregulation induced an increase in the cumulative mortality of WSSV-infected shrimp by reducing ROS production and suppressing the expression of antimicrobial peptides genes. The findings of present study suggest that LvROMO1 plays an important role in ROS production in L. vannamei and is involved in innate immunity.

Heat shock 70 kDa protein cognate 5 involved in WSSV toleration of Litopenaeus vannamei.

The expression levels of 97 unigenes encoding heat shock proteins of Litopenaeus vannamei was scanned, and ten of them were significantly induced by white spot syndrome virus (WSSV). Among these genes, heat shock 70 kDa protein cognate 5 (LvHSC70-5) was upregulated to the highest extent and subjected to further studies. Subcellular localization assay revealed that LvHSC70-5 was located in the mitochondria. Aside from WSSV infection, unfolded protein response activation and thermal stress could also upregulate LvHSC70-5. Results of reporter gene assay demonstrated that promoter of LvHSC70-5 was activated by L. vannamei heat shock factor protein 1, activating transcription factor 4 and thermal stress. A decrease in the expression of LvHSC70-5 could reduce the aggregation of proteins in hemocytes and the cumulative mortality of WSSV-infected L. vannamei. LvHSC70-5 in L. vannamei hemocytes was upregulated by mild thermal stress. In addition, mild thermal stress, decreased the copy number of WSSV in shrimp muscle and the cumulative mortality of WSSV-infected L. vannamei. Therefore, collecting results suggested that LvHSC70-5 should be involved in WSSV toleration of shrimp L. vannamei.

[Biomechanical study on a net-fixation of Kirschner wire in treating depressed tibial plateau fractures].

OBJECTIVE: To evaluate the biomechanical properties of tibial plateau depressed fracture fixed with a net-fixation of Kirschner wires. METHODS: Twenty homemade fracture models were fixed with eight 1.5 mm Kirschner wires in a net-fixation; 20 homemade fracture models were fixed with two 3.5 mm cortical screws. Plane-compressed and dot-compressed test were made on each 10 models of the two groups. The maximal force of anti-ompress and stiffness were measured and evaluated. RESULTS: In plane-compressed test,mean maximal force of anti-compress and stiffness for screw fixation was (1,925.31 +/- 444.26) N and (2.28 +/- 0.53) N/mm2, respectively, for net-fixation was (1,609.62 +/- 277.72) N and (1.90 +/- 0.33) N/mm2, respectively. There was no statistical difference between the two fixation methods (P > 0.05). In dot-compressed test,mean maximal force of anti-compress and stiffness for screw fixation was (411.13 +/- 233.88) N and (2.66 +/- 1.52) N/mm2,respectively,for net-fixation was (1,105.58 +/- 290.66) N and (7.18 +/- 1.89) N/mm2,respectively,the net-fixation was better than that of the screw fixation (P< 0.01). CONCLUSION: Treatment of tibial plateau depressed fracture with a net-fixation of Kirschner wires is a biological fixation and is a reliably method.

Overexpression of Insig-1 protects ß cell against glucolipotoxicity via SREBP-1c.

BACKGROUND: High glucose induced lipid synthesis leads to ß cell glucolipotoxicity. Sterol regulatory element binding protein-1c (SREBP-1c) is reported to be partially involved in this process. Insulin induced gene-1 (Insig-1) is an important upstream regulator of Insig-1-SREBPs cleavage activating protein (SCAP)-SREBP-1c pathway. Insig-1 effectively blocks the transcription of SREBP-1c, preventing the activation of the genes for lipid biosynthesis. In this study, we aimed to investigate whether Insig-1 protects ß cells against glucolipotoxicity. METHODS: An Insig-1 stable cell line was generated by overexpression of Insig-1 in INS-1 cells. The expression of Insig-1 was evaluated by RT-PCR and Western blotting, then, cells were then treated with standard (11.2 mM) or high (25.0 mM) glucose for 0 h, 24 h and 72 h. Cell viability, apoptosis, glucose stimulated insulin secretion (GSIS), lipid metabolism and mRNA expression of insulin secretion relevant genes such as IRS-2, PDX-1, GLUT-2, Insulin and UCP-2 were evaluated. RESULTS: We found that Insig-1 suppressed the high glucose induced SREBP-1c mRNA and protein expression. Our results also showed that Insig-1 overexpression protected ß cells from ER stress-induced apoptosis by regulating the proteins expressed in the IRE1α pathway, such as p-IRE1α, p-JNK, CHOP and BCL-2. In addition, Insig-1 up-regulated the expression of IRS-2, PDX-1, GLUT-2 and Insulin, down-regulated the expression of UCP-2 and improved glucose stimulated insulin secretion (GSIS). Finally, we found that Insig-1 inhibited the lipid accumulation and free fatty acid (FFA) synthesis in a time-dependent manner. CONCLUSIONS: There results suggest that Insig-1 may play a critical role in protecting ß cells against glucolipotoxicity by regulating the expression of SREBP-1c.

[Effect of sodium tungstate on glucose metabolism in adipocytes].

OBJECTIVE: To explore the effect of sodium tungstate on glucose metabolism in adipocytes and its mechanism. METHODS: After 3T3-L1 preadipocytes were differentiated into adipocytes, these adipocytes were incubated with sodium tungstate (0, 150, 300, 500, and 700 micromol/L) for 48 h, and then glucose consumption of the adipocytes was detected by glucose-oxidase assay. Glucose transport was determined by the uptake of 2-deoxy-[3H]-D-glucose, and the expression of glucose transport-4 (GLUT-4) mRNA was identified by semi-quantitative RT-PCR. RESULTS: Sodium tungstate (150 approximately 700 micromol/L) could significantly increase the glucose consumption and glucose transport with a concentration dependent-effect. Sodium tungstate could increase GLUT-4 mRNA expression. CONCLUSION: Sodium tungstate can enhance the glucose metabolism of adipocytes by up-regulating the expression of GLUT-4 mRNA.