Landform and lithospheric development contribute to the assembly of mountain floras in China.

Although it is well documented that mountains tend to exhibit high biodiversity, how geological processes affect the assemblage of montane floras is a matter of ongoing research. Here, we explore landform-specific differences among montane floras based on a dataset comprising 17,576 angiosperm species representing 140 Chinese mountain floras, which we define as the collection of all angiosperm species growing on a specific mountain. Our results show that igneous bedrock (granitic and karst-granitic landforms) is correlated with higher species richness and phylogenetic overdispersion, while the opposite is true for sedimentary bedrock (karst, Danxia, and desert landforms), which is correlated with phylogenetic clustering. Furthermore, we show that landform type was the primary determinant of the assembly of evolutionarily older species within floras, while climate was a greater determinant for younger species. Our study indicates that landform type not only affects montane species richness, but also contributes to the composition of montane floras. To explain the assembly and differentiation of mountain floras, we propose the 'floristic geo-lithology hypothesis', which highlights the role of bedrock and landform processes in montane floristic assembly and provides insights for future research on speciation, migration, and biodiversity in montane regions.

Hypothalamic NPFFR2 attenuates central insulin signaling and its knockout diminishes metabolic dysfunction in mouse models of diabetes mellitus.

BACKGROUND: The hypothalamus is a crucial brain region that mediates the effects of insulin and leptin signals on peripheral metabolic functions. Previous research has shown that insulin signals in the hypothalamus act via multiple neuronal circuits and anabolic/catabolic pathways that converge on the vagus nerve and sympathetic fibers to coordinate energy metabolism in peripheral organs. Additionally, neuropeptide FF (NPFF) has been identified as a regulator of feeding behaviors and energy homeostasis in the hypothalamus, but the mechanisms underlying its involvement in metabolic control remain unclear. This study aims to explore the underlying mechanisms of NPFF in modulating metabolic disorders. METHODS: In this study, we investigated the physiological role of NPFF in insulin-related energy homeostasis and metabolic health. First, we evaluated the effects of NPFF and its receptors on central insulin signaling using mouse hypothalamic cell lines and Npffr2-overexpressing mice. To further explore the effects of NPFFR2 on insulin-related metabolic disorders, such as diabetes mellitus, we used Npffr2-deleted mice in combination with the streptozotocin (STZ)-induced type 1 diabetes and high-fat diet/STZ-induced type 2 diabetic mouse models. The impacts of central NPFFR2 were demonstrated specifically through Npffr2 overexpression in the hypothalamic arcuate nucleus, which subsequently induced type 2 diabetes. RESULTS: We found that stimulating NPFFR2 in the hypothalamus blocked hypothalamic insulin activity. Npffr2 deletion improved central and peripheral metabolic symptoms in both mouse models of diabetes mellitus, exerting effects on central and systemic insulin resistance, feeding behaviors, glucose and insulin intolerance, lipid metabolism, liver steatosis, and inflammation of white adipose tissues. The overexpression of ARC Npffr2 augmented the metabolic dysregulation in the mouse model of type 2 diabetes. CONCLUSIONS: Our findings demonstrate that hypothalamic NPFFR2 negatively regulates insulin signaling in the central nervous system and plays an important role in maintaining systemic metabolic health, thereby providing valuable insights for potential clinical interventions targeting these health challenges.

Efficacy and safety of single Ultrasound Cyclo-Plasty to treat refractory glaucoma: Results at 1 year.

PURPOSE: To evaluate clinical efficacy and safety of single Ultrasound Cyclo-Plasty (UCP) in the treatment of advanced refractory glaucoma. METHODS: From January 2018 to August 2018, 25 patients (25 eyes) with refractory glaucoma and intraocular pressure (IOP) not controlled by drugs or conventional filtering surgery were included in the study. All subjects (neovascular glaucoma [n = 12], secondary glaucoma [n = 6], angle closure glaucoma [n = 6], and primary open angle glaucoma [n = 1]) underwent 8-sector Ultrasound Cyclo-Plasty. Patients were followed-up at Day 1, Week 1, and at 1, 3, 6, and 12 months, during which the IOP, the number of IOP lowering drugs and the occurrence of ocular complications were recorded. Clinical outcomes were IOP reduction, success rate, and ocular complications. According to the glaucoma type, patients were divided into a neovascular group (NVG) and a non-NVG group for sub-analysis. RESULTS: All patients underwent a single UCP procedure and mean IOP reduced significantly from 39.7 ± 6.1 mmHg before UCP to 27.1 ± 11.0 mmHg at 1 year (p < 0.01) corresponding to a mean IOP reduction of 29.6%. The mean number of IOP-lowering drugs used was 2.4 ± 1.2 at baseline and 2.3 ± 1.0 at 12 months. Success rate after a single UCP procedure was achieved in 41.7% patients at 1 year, with a higher success rate in non-NVG than in the NVG group. No major postoperative complications were reported. The main complication was conjunctival congestion, anterior chamber inflammation, scleral ring congestion, and scleral inprint. Of these, scleral ring congestion and scleral imprint are relatively rare complications, which can still be observed 12 months after UCP treatment. CONCLUSION: UCP for refractory glaucoma is effective in reducing IOP and has a good safety profile. Success rate is lower after a single UCP in NVG than for other types of glaucoma.

Fas/FasL of pacific cod mediated apoptosis.

Fas and Fas ligand (FasL) pathway plays important roles in virus defense and cell apoptosis. In our previous work, nervous necrosis virus (NNV) was discovered in Pacific cod (Gadus macrocephalus), and the Fas ligand (PcFasL) was up-regulated when NNV outbreak, however, signal transmission of Fas/FasL in fish are still unclear. In the present study, Pacific cod Fas (PcFas), PcFasL and Fas-associating protein with a novel death domain (PcFADD) were characterized. The predicted protein of PcFas, PcFasL and PcFADD includes 333 aa, 90 aa and 93 aa, separately. 3-D models of PcFas, PcFasL and PcFADD were well constructed based on reported templates, respectively, even though the sequence homology with other fish is very low. The transcript levels of PcFas increased gradually from 15 day-post hatching (dph) to 75dph. PcFas was significantly up-regulated when cod larvae had NNV symptoms at 24dph, 37dph, 46dph, 69dph, and 77dph. Subcellular localization revealed that PcFasL was located in the cytoplasm, while PcFas was mainly located in the cell membrane. Exogenous expressed PcFasL of 900 µg/mL could kill the Epithelioma papulosum cyprinid (EPC) cells by MTT test, but low concentration has no effect on the cells. qPCR analysis showed that overexpression of PcFas could significantly up-regulate the expression of genes related to Fas/FasL signaling pathway, including bcl-2, bax, and RIP3, while overexpression of PcFasL significantly up-regulate the expression of caspase-3, caspase-9, and MLKL. Overexpression of PcFas or PcFasL could induce EPC apoptosis significantly by flow cytometry, which was consistent with the results of caspase-3 mRNA level increasing. The results indicated that NNV could induce apoptosis through Fas/FasL signal pathway.

Clinical differences in chest CT characteristics between the progression and remission stages of patients with COVID-19 pneumonia.

INTRODUCTION: Computed tomography (CT) can be effective for the early screening and diagnosis of COVID-19. This study aimed to investigate the distinctive CT characteristics of two stages of the disease (progression and remission). METHODS: We included all COVID-19 patients admitted to Wenzhou Central Hospital from January to February, 2020. Patients underwent multiple chest CT scans at intervals of 3-10 days. CT features were recorded, such as the lesion lobe, distribution characteristics (subpleural, scattered or diffused), shape of the lesion, maximum size of the lesion, lesion morphology (ground-glass opacity, GGO) and consolidation features. When consolidation was positive, the boundary was identified to determine its clarity. RESULTS: The ratios of some representative features differed between the remission stage and the progression phase, such as round-shape lesion (8.0% vs 34.4%), GGO (65.0% vs 87.5%), consolidation (62.0% vs 31.3%), large cable sign (59.0% vs 9.4%) and crazy-paving sign (20.0% vs 50.0%). Using these features, we pooled all the CT data (n = 132) and established a logistic regression model to predict the current development stage. The variables consolidation, boundary feature, large cable sign and crazy-paving sign were the most significant factors, based on a variable named "prediction of progression or remission" (PPR) that we constructed. The ROC curve showed that PPR had an AUC of 0.882 (cutoff value = 0.66, sensitivity = 0.75, specificity = 0.875). CONCLUSION: CT characteristics, in particular, round shape, GGO, consolidation, large cable sign, and crazy-paving sign, may increase the recognition of the intrapulmonary development of COVID-19.

Effects of Anesthetic Technique on Postoperative Pulmonary Metastasis in Patients Undergoing Laryngectomy.

BACKGROUND: Whether laryngeal cancer is directly implanted into the lungs during orotracheal intubation is still unclear. Therefore, this study aimed to find whether orotracheal intubation is an independent risk factor for postoperative pulmonary metastasis in patients undergoing laryngectomy. PATIENTS AND METHODS: Medical records from January 1, 2006, to December 31, 2016, were reviewed. According to similar propensity scores, patients who received orotracheal intubation (tracheal intubation group, n = 515) were matched 1:1 with those who received tracheotomy (tracheotomy group, n = 326) in the induction of general anesthesia. The primary outcome was postoperative pulmonary metastasis. Secondary outcomes included local recurrence, lymphatic metastasis, tracheostomal recurrence and overall survival. RESULTS: Between the two groups, there was no significant difference in postoperative pulmonary metastasis (P = 0.688), local recurrence (P = 0.215), lymphatic metastasis (P = 0.480), tracheostomal recurrence (P = 0.246) or all-cause death (P = 0.299). The primary site of cancer was an independent risk factor for pulmonary metastasis [HR 0.29, 95% CI 0.13-0.68; P = 0.013] and local recurrence (HR 2.69, 95% CI 1.39-5.21; P = 0.003). Type of surgery (HR 3.13, 95% CI 2.03-4.84; P < 0.001) and N classification of TNM (HR 0.27, 95% CI 0.10-0.75; P = 0.012) were risk factors for local recurrence. Postoperative chemotherapy was an independent risk factor for lung metastasis (HR 7.58, 95% CI 3.11-18.47; P < 0.001) and lymphatic metastasis (HR 5.18, 95% CI 2.57-11.91; P < 0.001), and 5-year overall survival was associated with age (P = 0.028), clinical stage (P < 0.001) and postoperative chemotherapy (P = 0.003) but not with anesthetic technique (P = 0.473). CONCLUSION: This retrospective study suggests that orotracheal intubation in laryngectomy is not a risk factor for postoperative pulmonary metastasis, local recurrence, lymphatic metastasis or overall survival.

Identification of differentially expressed genes in gills of tiger puffer (Takifugu rubripes) in response to low-salinity stress.

Salinity is an important abiotic factor for aquatic organisms. In fish, changes in salinity affect physiological responses and alter the immune system. Takifugu rubripes is an important economic marine fish, and mechanisms of T. rubripes adaptation to salinity changes need to be further documented. In this study, a transcriptome sequencing technique was used to analyse genes that were differentially expressed in the T. rubripes gill after low-salinity stress for 30 d, and differential gene expression was further validated by quantitative real-time PCR (qPCR). After assembly, 385 differentially expressed genes (DEGs) were identified, including 182 upregulated genes and 203 downregulated genes. The DEGs were assigned to Gene Ontology (GO) classes with a total of 1647 functional terms. Most DEGs were assigned to biological process (984; 59.8%) followed by molecular function (445; 27.0%) and cellular component (218; 13.2%). Further KEGG analysis allocated 385 DEGs to 95 KEGG pathways. After q-value correction, 7 pathways (Glycolysis/Gluconeogenesis; Biosynthesis of amino acids; Carbon metabolism; Fructose and mannose metabolism; Pentose phosphate pathway; Metabolism of xenobiotics by cytochrome P450; and Glycine, serine and threonine metabolism) remained significant. qPCR results indicated that the transcripts of six selected genes sharply increased after 30 d of low-salinity stress. Low-salinity stress obviously increased SLC39A6, SLC5A9, NKAα1, CYP1A1, CYP1B1, and GSTA expression. In contrast, the genes encoding Aldoaa, GPI, FBP2 and GAPDH exhibited downregulation. In addition, three solute carrier (SLC) genes selected from the DEGs were further studied for differential expression patterns after low-salinity exposure, and the results showed that the SLCs were upregulated in T. rubripes after 72 h of low-salinity exposure. This investigation provides data for understanding the molecular mechanisms of fish responses to low-salinity stress and provides a reference for rationally setting salinity levels in aquaculture.

Conserved function of Pacific cod Caspase-3 in apoptosis.

Apoptosis plays a significant role in the cellular immune responses against infections, especially those related to viruses. Across various species, Caspase 3 is a prominent mediator of apoptosis and participates in the cell death signaling cascade. However, its role remains relatively unknown in cod fish. In this study, we aimed to reveal the role of Pacific cod Caspase-3 (GmCasp3) in apoptosis and its evolutionary position. Our results showed that the GmCasp3 cDNA contains an open reading frame of 864 nucleotides; that codes for 287 amino acids long protein with a molecular weight of 32.03 kDa. The sequence alignments and 3-D model indicated that GmCasp3 contained highly conserved domains, such as "QACRG", "GSWFI" and "HG" active sites, however, the phylogenetic tree analysis revealed that both GmCasp3 and Atlantic cod caspase-3 clustered together are far from the high vertebrate branch, indicating they are at a lower position in vertebrate evolution. Red fluorescent labeling vector pDsRed2-C1-GmCasp3 was constructed and it was transfected into EPC cell lines. The result showed that GmCasp3 protein was distribute in the protoplasm and expressed in apoptotic cell debris. Moreover, the GmCasp3 enzyme activity increased with the increased post-transfection analysis time, while the genome DNA was visibly fragmented at 36 h post transfection. Flow cytometry analysis showed that the proportion of apoptosis cells increased from 12 h to 24 h post transfection. In conclusion, the conserved functions of GmCasp3 in apoptosis indicated that Pacific cod has the similar apoptotic characteristics as other animals.

Prognostic value of time to generalization in 71 Chinese patients with sporadic amyotrophic lateral sclerosis / 中华医学杂志(英文版)

BACKGROUND@#It is important to determine prognostic factors for the outcome of amyotrophic lateral sclerosis (ALS) at an early stage. The time taken for symptoms to spread from spinal or bulbar regions to both (time to generalization; TTG) is considered a strong predictor of survival; however, this has rarely been studied in Asian populations. The aim of this retrospective study was to evaluate potential factors affecting prognosis in Chinese patients with sporadic ALS, with a focus on the association between TTG and overall survival.@*METHODS@#Seventy-one patients with sporadic ALS who were hospitalized at Chinese PLA General Hospital from 2009 to 2016 were followed up until December 2017. Survival analysis was performed using univariate Kaplan-Meier log-rank and multivariate Cox proportional hazards models. The clinical data of the patients were recorded and analyzed. Variables studied were age at symptom onset, sex, site of symptom onset, diagnostic latency, TTG, diagnostic category, ALS Functional Rating Scale-revised score, percent predicted forced vital capacity (FVC%), and disease progression rate (DPR) at diagnosis.@*RESULTS@#The mean age at onset was 54 (SD = 10.2) years, and the median survival time from symptom onset was 41 months (95% confidence interval: 34-47). By univariate analysis, factors independently affecting survival were age at symptom onset (Log rank = 15.652, P < 0.0001), TTG (Log rank = 14.728, P < 0.0001), diagnostic latency (Log rank = 11.997, P = 0.001), and DPR (Log rank = 6.50, P = 0.011). In the Cox multivariate model, TTG had the strongest impact on survival time (hazard ratio = 0.926, P = 0.01).@*CONCLUSIONS@#TTG can be used as an effective indicator of prognosis in patients with sporadic ALS.

Prognostic value of time to generalization in 71 Chinese patients with sporadic amyotrophic lateral sclerosis / 中华医学杂志(英文版)

Background:@#It is important to determine prognostic factors for the outcome of amyotrophic lateral sclerosis (ALS) at an early stage. The time taken for symptoms to spread from spinal or bulbar regions to both (time to generalization; TTG) is considered a strong predictor of survival; however, this has rarely been studied in Asian populations. The aim of this retrospective study was to evaluate potential factors affecting prognosis in Chinese patients with sporadic ALS, with a focus on the association between TTG and overall survival.@*Methods:@#Seventy-one patients with sporadic ALS who were hospitalized at Chinese PLA General Hospital from 2009 to 2016 were followed up until December 2017. Survival analysis was performed using univariate Kaplan-Meier log-rank and multivariate Cox proportional hazards models. The clinical data of the patients were recorded and analyzed. Variables studied were age at symptom onset, sex, site of symptom onset, diagnostic latency, TTG, diagnostic category, ALS Functional Rating Scale-revised score, percent predicted forced vital capacity (FVC%), and disease progression rate (DPR) at diagnosis.@*Results:@#The mean age at onset was 54 (SD = 10.2) years, and the median survival time from symptom onset was 41 months (95% confidence interval: 34–47). By univariate analysis, factors independently affecting survival were age at symptom onset (Log rank = 15.652, P < 0.0001), TTG (Log rank = 14.728, P < 0.0001), diagnostic latency (Log rank = 11.997, P = 0.001), and DPR (Log rank = 6.50, P = 0.011). In the Cox multivariate model, TTG had the strongest impact on survival time (hazard ratio = 0.926, P = 0.01).@*Conclusions:@#TTG can be used as an effective indicator of prognosis in patients with sporadic ALS.

Transcriptome from Pacific cod liver reveals types of apolipoproteins and expression analysis of AFP-IV, structural analogue with mammalian ApoA-I.

Apolipoproteins (Apos), transporting the lipids through the lymphatic and circulatory systems, are associated with kinds of diseases. Additionally, type IV antifreeze protein (AFP-IV) was related evolutionarily with apolipoproteins. However, the information of Apos in fish was limited. In this study, ApoA-I, ApoA-I-2, ApoA-IV, Apo E, ApoB-100-like and AFP-IV were sequenced from Pacific cod (Gadus macrocephalus) liver transcriptome using Illumina HiSeq 2000, and their 3-D models were constructed based on the most confidence templates ever reported in mammals. Interestingly, the model of G. macrocephalus AFP-IV, named GmAFPIV, is quite similar to the structure of ApoA-I. GmAFPIV includes 689 bases with a complete open reading frame encoding 125 amino acids. Sequence alignment of GmAFPIV showed 30% to 50% similarity with that of other species except Gadus sp. Expression levels of GmAFPIV were found in a decreasing manner in liver, intestine, gill, brain and gonad. Heterologously expression of the GmAFPIV protein was expressed in Escherichia coli and purified to immunize New Zealand rabbits. The survivors of E. coli in 60 µg/mL of GmAFPIV are more than that in the 30 µg/mL group after stored in -20 °C and -80 °C, indicating high concentration of GmAFPIV could protect E. coli avoiding the damage from ice crystal. The subcellular localization of GmAFPIV showed that the green fluorescence was mainly observed in the cytoplasm, indicating GmAFPIV play roles in the cytoplasm. It was concluded that GmAFPIV may function not only as an antifreeze protein but also as an apolipoprotein transporting lipids in fish.

Molecular characterization of caspase members and expression response to Nervous Necrosis Virus outbreak in Pacific cod.

Multiple functions of caspases include normal cell turnover, proper development and function of the immune system, hormone-dependent atrophy, embryonic development and chemical-induced cell injury. During artificial propagation of Pacific cod, Gadus macrocephalus, high mortality occurred during early development stages. Here, we performed various analyses on the cDNA and protein sequences of six different G. macrocephalus caspases namely GmCasp3, 6, 7, 8, 9 and 10, and tried to investigate the contributions of caspase family to the development and Nervous Necrosis Virus (NNV) resistance. Sequence analysis of GmCaspase proteins showed that each caspase shared conserved domains like "HG", "QACXG (X for R, G or Q)" and "GSWF" except GmCasp10. Sequence alignment and phylogenetic tree showed that GmCasp8 and GmCasp10 were quite different from those of other fishes. 3-D models indicated that structure of GmCasp3 is very conservative, but GmCasp6, 7, 8, 9 and 10 are less conservative. Tissue distribution analysis showed that six Gmcaspases mRNA transcripts were detected in tissues of intestine, gill, thymus, head-kidney and spleen with different abundance, but Gmcasp7 were not detected in the brain. GmCasp3 transcript was kept at very low level in the early development stages, while the expression levels of GmCasp6, 7, 8, 10 were different at various development stages. GmCasp8 level seemed to be much higher than other caspases in the heads of 65dph and 75dph juveniles. To understand the role of caspases during NNV outbreak, modulation in expression of each Gmcaspases were investigated. The results showed that GmCasp3 transcript level increased significantly when NNV broke out, while GmCasp7, 8, 9 and 10 in cod heads decreased obviously at 69dph and 77dph. The results suggest that caspases in Pacific cod should be diverse in their structure and function, and their unique features and response to NNV outbreak add more evidences for the specificity of immune system in Pacific cod.

Digital gene expression analysis of Takifugu rubripes brain after acute hypoxia exposure using next-generation sequencing.

The adverse effects of hypoxia are confined to biochemical, physiological, developmental and behavioral processes, especially injury of the brain. In this study, a subset of genes in the brain of Takifugu rubripes were analyzed using digital gene expression (DGE) profiles and next-generation sequencing after acute hypoxia. Among 32 differentially expressed genes, 29 were up-regulated and 3 were down-regulated following hypoxia exposure. Using Gene Ontology analysis, it was found that transcription and translation, metabolism, and the stress response were affected by exposure to hypoxia. KEGG analysis revealed that the neuroactive ligand-receptor interaction pathway was significantly enriched in hypoxia-exposed T. rubripes. To further confirm the differential expression of genes, quantitative real-time PCR was performed to test six candidate genes, with the following five genes exhibiting the same expression patterns as the sequencing results: Proto-oncogene c-fos, Kruppel-like factor 2, immediate early response 2, proopiomelanocortin A and rhodopsin. This work is the first to identify and annotate genes in T. rubripes affected by hypoxia stress. This investigation provides data for understanding the molecular mechanism of fish adaptation to hypoxia and provides a reference for rationally setting dissolved oxygen levels in aquaculture.

Efficient therapeutic delivery by a novel cell-permeant peptide derived from KDM4A protein for antitumor and antifibrosis.

Cell-penetrating peptide (CPP) based delivery have provided immense potential for the therapeutic applications, however, most of nonhuman originated CPPs carry the risk of possible cytotoxicity and immunogenicity, thus may restricting to be used. Here, we describe a novel human-derived CPP, denoted hPP10, and hPP10 has cell-penetrating properties evaluated by CellPPD web server, as well as In-Vitro and In-Vivo analysis. In vitro studies showed that hPP10-FITC was able to penetrate into various cells including primary cultured cells, likely through an endocytosis pathway. And functionalized macromolecules, such as green fluorescent protein (GFP), tumor-specific apoptosis inducer Apoptin as well as biological active enzyme GCLC (Glutamate-cysteine ligase, catalytic subunit) can be delivered by hPP10 in vitro and in vivo. Collectively, our results suggest that hPP10 provide a novel and versatile tool to deliver exogenous proteins or drugs for clinical applications as well as reprogrammed cell-based therapy.

Evidence for and characterization of nervous necrosis virus infection in Pacific cod (Gadus macrocephalus).

A mortality rate higher than 90% was observed in a larva-rearing facility for Pacific cod, Gadus macrocephalus, in China. Larvae showing clinical signs of infection were collected. Initial suspicion of nervous necrosis virus (NNV) infection was confirmed by sequencing, absolute quantification real-time PCR (A-qPCR), and electron microscopy. The nucleotide sequence of RNA2 was 1,375 bases long (GenBank no. KM576685), coding for a single ORF corresponding to the capsid protein from residues 21 to 1034. Phylogenetic analysis of the capsid protein sequence showed that PCNNV belongs to the barfin flounder NNV (BFNVV) genotype. An amino acid sequence alignment revealed 39 differences between the cold- and warm-resistant viral groups, suggesting that PCNNV evolved under temperature selection. The 3-D structure of the predicted capsid protein was modeled to identify potential epitopes, and the gene was expressed in Escherichia coli, yielding a protein with a molecular mass of 55 kDa. During PCNNV outbreaks, the viral copy number was found to reach 10(7) per ng of total RNA, which could be considered the lethal copy number of NNV in cod. The gonads, eggs, fertilized eggs and asymptomatic cod fry were all positive for PCNNV, indicating viral vertical transmission as the main source of the viral load. The amount of virus in the apparent healthy fry or survivors seemed to decrease gradually with development. These results might lead to efficient diagnostic methods to help farmers select NNV-free broodfish for cod breeding.

Enhanced Peptide delivery into cells by using the synergistic effects of a cell-penetrating Peptide and a chemical drug to alter cell permeability.

Cell-penetrating peptides (CPPs) are short, often hydrophilic peptides that can deliver many kinds of molecules into cells and that are likely to serve as a useful tool of future biotherapeutics. However, CPPs application is limited because of insufficient transduction efficiency and unpredictable cellular localization. Here, we investigated the enhancement of 1,2-benzisothiazolin-3-one (BIT) on the uptake of a synthetic fluorescent TAT and a TAT-conjugated green fluorescent protein (GFP) or pro-apoptotic peptide KLA and evaluated its toxicity in various cell lines. Our results showed that BIT pretreatment can enhance the penetration efficiency of TAT and its fusion peptide. In addition, the fluorescence of the peptide conjugate at effective doses was well-distributed in the intracellular of different cell lines without membrane perforation or detectable cytotoxicity. The internalization of the peptides was serum-dependent and temperature-independent. These findings imply that BIT may serve as a newly found delivery enhancer that is suitable for improving the penetration of CPPs.

Transcriptomic analysis and biomarkers (Rag1 and Igµ) for probing the immune system development in Pacific cod, Gadus macrocephalus.

Mortality (>90%) is a big concern in larval rearing facilities of Pacific cod, Gadus macrocephalus, limiting its culture presently still in the experimental stages. Understanding the immune system development of G. macrocephalus is crucial to optimize the aquaculture of this species, to improve the use of economic resources and to avoid abuse of antibiotics. For the transcriptome analysis, using an Illumina sequencing platform, 61,775,698 raw reads were acquired. After a de novo assembly, 77,561 unigenes were obtained. We have classified functionally these transcripts by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). 27 genes mainly related to hematopoietic or lymphoid organ development and somatic diversification of immune receptors have been reported for the first time in Pacific cod, and 14 Ig heavy chain (µ chain) locuses were assembled using Trinity. Based on our previous achievement, we have chosen Rag1 and Igµ as immune system development biomarkers. Full length cDNA of Rag1 and Igµ as biomarkers were obtained respectively using RACE PCR. Concerning Rag1, the deduced amino acid of Rag1 and protein immunodetection revealed a Rag1 isoform of 69 kDa, significantly different from other fish orthologs, such as Oncorhynchus mykiss (121 kDa). Phylogenetic analysis reveals a unique immune system for the Gadus genre, not exclusive for Atlantic cod, among vertebrates. Meanwhile, full length cDNA of Igµ included an ORF of 1710 bp and the deduced amino acid was composed of a leader peptide, a variable domain, CH1, CH2, Hinge, CH3, CH4 and C-terminus, which was in accordance with most teleost. Absolute quantification PCR revealed that significant expression of Rag1 appeared earlier than Igµ, 61 and 95 dph compared to 95 dph, respectively. Here we report the first transcriptomic analysis of G. macrocephalus as the starting point for genetic research on immune system development towards improving the Pacific cod aquaculture.

Clinical analysis on juvenile dermatomyositis combined with soft-tissue calcification / 中国医师进修杂志

Objective To analyze the clinical features of juvenile dermatomyositis (JDM) combined with soft-tissue calcification. Methods Forty-seven patients with JDM combined with soft-tissue calcification (soft-tissue calcification group) were retrospectively analyzed, and they were contrasted with 89 patients with non-calcification (non-calcification group). Results The rates of Gotton signe, muscle contracture and joint dysfunction in soft-tissue calcification group were significantly higher than those in non-calcification group:87.23% (41/47) vs. 43.82% (39/89) and 68.09% (32/47) vs. 21.35% (19/89), and there were statistical differences (P<0.05). The dosage of glucocorticoid (conversion of prednisone measuring more than 1.5 mg/kg), rate of using immunodepressant, level of creatine kinase in soft-tissue calcification group were significantly lower than those in non-calcification group:17.02%(8/47) vs. 68.54%(61/89), 25.53%(12/47) vs. 88.76%(79/89), (566.45±240.41) U/L vs. (1 680.12±656.50) U/L, and there were statistical differences (P<0.05). Conclusions The patients with JDM combined with Gotton signe are more prone to soft-tissue calcification. The rate of muscle contracture and joint dysfunction in soft-tissue calcification patients is significantly higher than that in non-calcification patients. For the patients whose creatine kinase are not obviously elevated, they are more prone to soft-tissue calcification. Early active application of glucocorticoid and immunodepressant therapy can reduce or prevent the occurrence or development of late calcium deposition.

Percutaneous Transforaminal Endoscopic Spine System in Lumbar Discectomy for Disc Herniation / 天津医药

Objective To investigate the value of percutaneous transforaminal endoscopic spine system (TESSYS) in lumbar discectomy for disc herniation. Methods Thirty one patients with lumbar disc herniation were treated with TES-SYS and followed up 6-12 months. The involved levels of vertebral segment included L34 (2 cases), L45 (21 cases) and L5S1 (8 cases). The targeted puncture was performed under local anesthesia and fluoroscopic guidance. The foramen of involved level of vertebral segment was enlarged gradually with four trephinations, and the working cannula was inserted transforaminal in-to the canal. Then the herniation was exposed and removed with full endoscopic technique, including the loosen nucleus pulposus. The nerve root and dural sac were exposed and released adequately. Results The procedure was evenly carried out in 27 cases. After discectomy, the nerve roots were complete released, and not exposed in the first case of far lateral herni-ation and the second case of central herniation. The third case and eleventh case converted to microendoscopic discectomy, due to large herniation and intraoperative pain, respectively. The patients could walk in the same day, 1 or 2 days after opera-tion, with obvious relief of leg pain. One case of recurrence was found at 2 weeks after operation, who was treated conserva-tively. At the final follow-up, the visual analogue scale of leg pain decreased from 8.1±1.9 to 1.1±0.9, and the Oswestry dis-ability index (ODI) decreased from 31.1±8.3 to 3.4±3.3. According to MacNab scale, there were excellent results in 25 cases and good results in 6 cases. Conclusion The percutaneous endoscopic TESSYS is a good minimal invasive technique for lumbar discectomy, with good results and a learning curve.