[Near-infrared targeted probe designed for intraoperative imaging of prostatic neurovascular bundles].

OBJECTIVE: To investigate the imaging effect of a near-infrared fluorescent targeted probe ICG-NP41 on the neurovascular bundles (NVB) around the prostate in rats. METHODS: A near-infrared fluorescent targeted probe ICG-NP41 was synthesized. An animal model for NVB imaging was established using Sprague-Dawley rats (250-400 g). Experiments were conducted using a custom-built near-infrared windowⅡ(NIR-Ⅱ) small animal in vivo imaging system, and images collected were processed using ImageJ and Origin. The fluorescence signal data were statistically analyzed using GraphPad Prism. The signal-to-background ratio (SBR) for NVB was quantitatively calculated to explore the effective dosage and imaging time points. Finally, paraffin pathology sections and HE staining were performed on the imaging structures. RESULTS: Except for rats in the control group (n=2), right-sided NVB of the rats injected with ICG-NP41 (n=2 per group) were all observed in NIR-Ⅱ fluorescence mode 2 h and 4 h after administration. At 2 h and 4 h, average SBR of cavernous nerve in 2 mg/kg group in fluorescence mode was 1.651±0.142 and 1.619±0.110, respectively, both higher than that in white light mode (1.111±0.036), with no significant difference (P>0.05); average SBR of 4 mg/kg group in fluorescence mode were 1.168±0.066 and 1.219±0.118, respectively, both higher than that in white light mode (1.081±0.040), with no significant difference (P>0.05). At 2 h and 4 h, the average SBR of 2 mg/kg and 4 mg/kg groups in fluorescence mode were higher than that of the control group (SBR=1), the average SBR of the 2 mg/kg group was higher than that of the 4 mg/kg group, and all the above with no significant difference (P>0.05). The average diameter of the nerve measured by full width at half maxima method was about (178±15) µm. HE staining of paraffin sections showed the right major pelvic ganglion. CONCLUSION: The near-infrared fluorescent targeted probe ICG-NP41 can be used for real-time imaging of the NVB around the prostate in rats, providing a potential feasible solution for localizing NVB in real time during nerve-sparing radical prostatectomy.

Neutrophil extracellular traps participate in the development of cancer-associated thrombosis in patients with gastric cancer.

BACKGROUND: The development of venous thromboembolism (VTE) is associated with high mortality among gastric cancer (GC) patients. Neutrophil extracellular traps (NETs) have been reported to correlate with the prothrombotic state in some diseases, but are rarely reported in GC patients. AIM: To investigate the effect of NETs on the development of cancer-associated thrombosis in GC patients. METHODS: The levels of NETs in blood and tissue samples of patients were analyzed by ELISA, flow cytometry, and immunofluorescence staining. NET generation and hypercoagulation of platelets and endothelial cells (ECs) in vitro were observed by immunofluorescence staining. NET procoagulant activity (PCA) was determined by fibrin formation and thrombin-antithrombin complex (TAT) assays. Thrombosis in vivo was measured in a murine model induced by flow stenosis in the inferior vena cava (IVC). RESULTS: NETs were likely to form in blood and tissue samples of GC patients compared with healthy individuals. In vitro studies showed that GC cells and their conditioned medium, but not gastric mucosal epithelial cells, stimulated NET release from neutrophils. In addition, NETs induced a hypercoagulable state of platelets by upregulating the expression of phosphatidylserine and P-selectin on the cells. Furthermore, NETs stimulated the adhesion of normal platelets on glass surfaces. Similarly, NETs triggered the conversion of ECs to hypercoagulable phenotypes by downregulating the expression of their intercellular tight junctions but upregulating that of tissue factor. Treatment of normal platelets or ECs with NETs augmented the level of plasma fibrin formation and the TAT complex. In the models of IVC stenosis, tumor-bearing mice showed a stronger ability to form thrombi, and NETs abundantly accumulated in the thrombi of tumor-bearing mice compared with control mice. Notably, the combination of deoxyribonuclease I, activated protein C, and sivelestat markedly abolished the PCA of NETs. CONCLUSION: GC-induced NETs strongly increased the risk of VTE development both in vitro and in vivo. NETs are potential therapeutic targets in the prevention and treatment of VTE in GC patients.

[Mechanism of Shenxiong Glucose Injection against myocardial ischemia-reperfusion injury based on network pharmacology and molecular docking].

Based on network pharmacology and molecular docking, the mechanism of danshensu and tetramethylpyrazine, the main active components of Shenxiong Glucose Injection(SGI), against myocardial ischemia-reperfusion injury(MIRI) was explored. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), GeneCards, and Online Mendelian Inheri-tance in Man(OMIM) were used to search the targets of the active components and the disease, and the common targets were screened. The "drug-component-disease-target" network was constructed by Cytoscape, and the protein-protein interaction network was established by STRING, followed by Gene Ontology(GO) term and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment by R software. AutoDock Vina was employed for the molecular docking between active components and core targets. A total of 15 potential targets of danshensu and tetramethylpyrazine against MIRI were screened out, involving the major GO terms of cyclooxyge-nase pathway, extracellular matrix binding, and antioxidant activity, and the main pathways of platelet activation and regulation of lipolysis in adipocytes. Danshensu and tetramethylpyrazine can form stable conformations with core targets prostaglandin G/H synthase 2(PTGS2), vascular endothelial growth factor A(VEGFA), and acetylcholinesterase(ACHE) with low binding energy. This study reflects the multi-component, multi-target, multi-pathway, and synergistic action characteristics of SGI, which provides a theoretical re-ference for further clarifying the anti-MIRI mechanism of SGI.

Exogenous glutathione exerts a therapeutic effect in ischemic stroke rats by interacting with intrastriatal dopamine.

We previously showed that oral administration of exogenous glutathione (GSH) exerted a direct and/or indirect therapeutic effect on ischemic stroke rats, but the underlying mechanisms remain elusive. In the current study, we conducted a quantitative proteomic analysis to explore the pathways mediating the therapeutic effect of GSH in cerebral ischemia/reperfusion (I/R) model rats. Rats were subjected to middle cerebral artery occlusion (MCAO) for 2 h followed by reperfusion. The rats were treated with GSH (250 mg/kg, ig) or levodopa (L-dopa, 100 mg/kg, ig) plus carbidopa (10 mg/kg, ig). Neurologic deficits were assessed, and the rats were sacrificed at 24 h after cerebral I/R surgery to measure brain infarct sizes. We conducted a proteomic analysis of the lesion side striatum samples and found that tyrosine metabolism and dopaminergic synapse were involved in the occurrence of cerebral stroke and the therapeutic effect of GSH. Western blot assay revealed that tyrosine hydroxylase (TH) mediated the occurrence of I/R-induced ischemic stroke and the therapeutic effect of GSH. We analyzed the regulation of GSH on endogenous small molecule metabolites and showed that exogenous GSH had the most significant effect on intrastriatal dopamine (DA) in I/R model rats by promoting its synthesis and inhibiting its degradation. To further explore whether DA-related alterations were potential targets of GSH, we investigated the therapeutic effect of DA accumulation on ischemic brain injury. The combined administration of the precursor drugs of DA (L-dopa and carbidopa) significantly ameliorated neurological deficits, reduced infarct size, and oxidative stress, and decreased pro-inflammatory cytokines levels in the striatum of I/R injury rats. More interestingly, exogenous L-dopa/carbidopa could also greatly enhance the exposure of intracerebral GSH by upregulating GSH synthetases and enhancing homocysteine (HCY) levels in the striatum. Thus, administration of exogenous GSH exerts a therapeutic effect on ischemic stroke by increasing intrastriatal DA, and the accumulated DA can, in turn, enhance the exposure of GSH and its related substances, thus promoting the therapeutic effect of GSH.

[Clinical features of children with severe adenovirus pneumonia and hemophagocytic syndrome: an analysis of 30 cases].

OBJECTIVE: To study the clinical features of children with severe adenovirus pneumonia (SAP) and hemophagocytic syndrome (HPS). METHODS: A retrospective analysis was performed from the chart review data of 30 children with SAP and HPS who were admitted from January 2014 to June 2019. According to the prognosis, the children were divided into a good prognosis group (n=18) and a poor prognosis group (n=12). RESULTS: Among the 30 children with SAP and HPS, the ratio of male to female was 2:1. The median age of onset was 1 year and 3 months (range 3 months to 5 years), and the mean course of fever was 19±7 d. Of the 30 children, 28 (93%) experienced disease onset in January to June. High-throughput gene detection of serum pathogens showed that 16 (53%) children were positive for human adenovirus type 7 (HAdV-7), and the other 14 (47%) children were positive for HAdV antigen based on immunofluorescence assay for throat swab, with unknown type. Of all 30 children, 29 (97%) had respiratory complications, 24 (80%) had cardiovascular complications, 16 (53%) had gastrointestinal complications, and 9 (30%) had toxic encephalopathy. Eighteen children (60%) improved or recovered and 12 (40%) did not recover (3 died). Compared with the good prognosis group, the poor prognosis group had a significantly longer course from onset to diagnosis of HPS (P<0.05), significantly higher levels of fibrinogen and tumor necrosis factor-α (P<0.05), and a significantly lower level of interferon-γ (P<0.05). The mean follow-up time was 6±2 months; 11 (41%) children recovered, 1 (4%) experienced recurrence of HPS, and 15 (56%) had the sequela of post-infectious bronchiolitis obliterans (PIBO). CONCLUSIONS: HPS may be observed in children with SAP, and PIBO is the most common sequela of SAP.

Combining antioxidant astaxantin and cholinesterase inhibitor huperzine A boosts neuroprotection.

Oxidative stress is a pathophysiological condition resulting in neurotoxicity, which is possibly associated with neurodegenerative disorders. In this study, the antioxidative effects of the antioxidant astaxanthin (AXT) in combination with huperzine A (HupA), which is used as a cholinesterase inhibitor for the treatment of Alzheimer's disease, were investigated. PC12 cells were treated with either tert­butyl hydroperoxide (TBHP), or with the toxic version of ß­amyloid, Aß25­35, to induce oxidative stress and neurotoxicity. Cell viability, morphology, lactate dehydrogenase (LDH) release, intracellular accumulation of reactive oxygen species (ROS), superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were determined, while neuroprotection was also monitored using an MTT assay. It was found that combining AXT with HupA significantly increased the viability of PC12 cells, prevented membrane damage (as measured by LDH release), attenuated intracellular ROS formation, increased SOD activity and decreased the level of MDA after TBHP exposure when compared to these drugs administered alone. Pretreatment with HupA and AXT decreased toxic damage produced by Aß25­35. These data indicated that combining an antioxidant with a cholinesterase inhibitor increases the degree of neuroprotection; with future investigation this could be a potential therapy used to decrease neurotoxicity in the brain.

Co-delivery of cisplatin and doxorubicin by covalently conjugating with polyamidoamine dendrimer for enhanced synergistic cancer therapy.

Because of the synergistic effects of drugs and minimal drug dose for cancer therapy, combination chemotherapy is frequently used in the clinic. In this study, hyaluronic acid-modified amine-terminated fourth-generation polyamidoamine dendrimer nanoparticles were synthesized for systemic co-delivery of cisplatin and doxorubicin (HA@PAMAM-Pt-Dox). In vitro data showed that HA@PAMAM-Pt-Dox can enter the cells through the lysosome mediated-pathway in a time-dependent manner. Cell viability studies indicated that HA@PAMAM-Pt-Dox exhibited a higher anticancer activity on MCF-7 and MDA-MB-231 breast cancer cells at a relative low concentration. HA@PAMAM-Pt-Dox not only efficiently inhibited tumor growth but also significantly reduced the toxicity of Dox. Moreover, intravenous administration of HA@PAMAM-Pt-Dox to MDA-MB-231 tumor-bearing BALB/c nude mice resulted in the accumulation of HA@PAMAM-Pt-Dox at the tumor site, thereby significantly inhibiting tumor growth without apparent toxicity. These results suggested that HA@PAMAM-Pt-Dox has great potential to improve the chemotherapeutic efficacy of cisplatin and doxorubicin in breast cancer. STATEMENT OF SIGNIFICANCE: One of the main problems in cancer treatment is the development of drug resistance. To date, it is believed that combination chemotherapy might be an effective strategy for the above problem. However, for two completely different drugs, combination chemotherapy faces huge difficulties including the antagonistic nature of drugs, variations in drugs in terms of solubility, and limited tumor targeting. Recent developments in nanoscience and nanotechnology provide an effective approach for such disadvantages. Considering the advantages of dendrimers such as control of size and molecular weight, bioavailability, and biosafety, we used fourth-generation dendrimers modified by HA as drug vectors by covalently conjugating them with anticancer drugs (cisplatin and doxorubicin) to form a nanodrug delivery system, named HA@PAMAM-Pt-Dox. We observed that the HA@PAMAM-Pt-Dox system can effectively kill breast cancer cells both in vitro and in vivo, which showed a favorable synergistic effect. This strategy can be extended to other drugs, thus providing a highly effective strategy for cancer treatment.

[Comparison of stripping of steep sheep maxillary sinus mucosal with new-type memory elastic silk sleeve stripper and umbrella stripper].

OBJECTIVE: This work aims to investigate the effect of stripping of steep sheep maxillary sinus by using 0.012 or 0.014 model new-type memory elastic silk sleeve stripper and umbrella stripper. METHODS: Goats with sinus floor gradients of 60° to 90° were selected by computed tomography of sheep head. A total of 72 animal models were established and randomized into three groups (n=24): group A (0.012 model), B (0.014 model) and C (umbrella stripper). Alveolar and maxillary sinus mucosa were stripped after crowning, and stripping length was measured when the stripping limit was reached or mucosal perforation occurred. RESULTS: The average stripping length of mucosa in group A was 11.3 mm±4.6 mm, and three cases experienced perforation of sinus floor mucosa. The average stripping length of mucosa in group B was 17.5 mm±5.0 mm, and one case experienced perforation of sinus floor mucosa. The average stripping length of mucosa in group C was 4.2 mm±1.3 mm, and four cases experienced perforation of sinus floor mucosa. The difference among the three groups was statistically significant (P<0.01) according to variance analysis. Moreover, the comparison between any two means was also statistically significant according to Dunnett's T3 test (P<0.05). CONCLUSIONS: The new-type memory elastic silk sleeve stripper effectively stripped steep maxillary sinus mucosa. The 0.014 model exhibited superior peeling effect and was relatively safe.

Identification of differentially expressed proteins in the gastric mucosal atypical hyperplasia tissue microenvironment.

In the present study, the interaction of proteins in the microenvironment of gastric mucosal atypical hyperplasia was analyzed. The stromata of normal gastric mucosa (NGM) and gastric mucosal atypical hyperplasia (GMAH) tissues were purified with laser capture microdissection (LCM). The differentially expressed GMAH proteins of the NGM and GMAH tissues were identified by quantitative proteomic techniques with isotope labeling. The cross-talk between differentially expressed proteins in NGM and GMAH tissues was then analyzed by bioinformatics. There were 165 differentially expressed proteins identified from the stromata of NGM and GMAH tissues. Among them, 99 proteins were upregulated and 66 were downregulated in GMAH tissue. The present study demonstrated that these proteins in gastric mucosal atypical hyperplasia were involved in cancer-associated signaling pathways, including the p53, mitogen-activated protein kinase (MAPK), cell cycle and apoptosis signaling pathways, and were involved in cellular growth, cellular proliferation, apoptosis and the humoral immune response. The results of the present study suggest that the 165 differentially expressed proteins, including S100 calcium-binding protein A6 (S100A6) and superoxide dismutase 3 (SOD3) in the microenvironment of gastric mucosal atypical hyperplasia, are involved in the p53, MAPK, cell cycle and apoptosis signaling pathways, and serve a function in the pathogenesis of gastric cancer.

[Cloning, expression and immuno-protection analysis of a gene encoding troponin T of Schistosoma japonicum (SjTnT)].

OBJECTIVE: To clone cDNA encoding troponin T of Schistosoma japonicum (SjTnT), and evaluate the protective efficacy induced by recombinant SjTnT in BALB/c mice against S. japonicum challenge infection. METHODS: The SjTnT gene was amplified from 28-day-schistosome cDNAs by PCR and then subcloned into pET28a(+). The recombinant SjTnT protein (rSjTnT) was expressed in Escherichia coli BL21 (DE3) cells. The serum specific to rSjTnT was prepared by immunized BALB/c mice with the recombinant antigen, and the immunogenicity of rSjTnT was detected by Western blotting and ELISA. The immuno-protective efficacy induced by rSjTnT in BALB/c mice was evaluated according to the reduction in worm and egg counts. RESULTS: The cDNA encoding SjTnT was successfully cloned and expressed in E. coli. Western blotting showed that rSjTnT had a good immunogenicity. The high level of specific IgG antibodies was detected, and 33.89% worm reduction and 43.94% liver egg reduction were obtained in mice vaccinated with rSjTnT combined with Seppic 206 adjuvant compared with those in the adjuvant control group. CONCLUSIONS: rSjTnT could induce partial immuno-protection against S. japonicum infection in BALB/c mice. This study provided a basic for understanding the biological function of SjTnT.

Seasonal variations in the energy budget of Elliot's pheasant (Syrmaticus ellioti) in cage.

This study aimed to discuss the energy budget of Elliot's pheasant Syrmaticus ellioti in different seasons, with life and health, good growth and normal digestion of Elliot's pheasant as the tested objects, The energy budget of Elliot's pheasant was measured by daily collection of the trial pheasants' excrement in the biological garden of Guangxi Normal University from March 2011 to February 2012. The results showed that the gross energy consumption, metabolic energy and excrement energy varied by season, increasing as temperature decreased. There was significant difference in gross energy consumption, metabolic energy, excrement energy between adults and nonages. There was also a trend that food digestibility of pheasants increases as temperature increases. In the same season, the food digestibility of adults was better than that of nonages. Throughout spring, summer, autumn and winter, the metabolic energy of 4-year adults were 305.77±13.40 kJ/d, 263.67±11.89 kJ/d, 357.23±25.49 kJ/d and 403.12±24.91 kJ/d, respectively, and the nonages were 284.86±17.22 kJ/d, 284. 66±15.16 kJ/d, 402. 26±31.46 kJ/d and 420. 30±31.98 kJ/d, respectively. The minimum metabolic energies were 247.65±21.81 g, 265.86±26.53 g, respectively for each group, detected between 4-year adults and 1-year nonages. Further study is needed to determine whether 29.6 C is the optimal temperature for the Elliot's pheasant.

Metabolism and thermoregulation between Mrs Hume's Pheasant (Syrmaticus humiae) and Elliot's Pheasant (S. ellioti).

To understand metabolic adaptations, the basal metabolic rate (BMR) of Mrs Hume's Pheasant (Syrmaticus humiae) and Elliot's Pheasant (Syrmaticus ellioti) were investigated. Metabolic rate (MR), body temperature (T(b) ) and thermal conductance (C) were determined in both species at a temperatrue range of 5-35 Degrees Celsius, respectively. Oxygen consumption was measured with a closed circuit respirometer. The thermal neutral zones (TNZ) were 24.5-31.6 Degrees Celsius, and 23.0-29.2 Degrees Celsius, respectively. With a temperature range of 5-35 Degrees Celsius, Mrs Hume's Pheasant and Elliot's Pheasant could maintained stable T(b) at a mean of (40.47 ± 0.64) and (40.36 ± 0.10) Degrees Celsius, respectively. Mean BMRs within TNZs were (1.36 ± 0.84) mLO2/(g.h) for Mrs Hume's Pheasant and (2.03 ± 0.12) mLO2/(g.h) for Elliot's Pheasant, which were 77% and 86% of the expected value based on their body mass, respectively. Thermal conductance of Mrs Hume's Pheasant and Elliot's Pheasant were (0.12 ± 0.01) and (0.17 ± 0.01) mLO2/(g.h.Degrees Celsius), below the lower critical temperature, respectively, which were 119% and 124% of the expected value based on their body mass, respectively. The ecophysiological characteristics of these species were low metabolic rate, high body temperature, and high thermal conductance, which allow both species to better adapt to the warmer climate environment in south China.

Molecular characterization and functional commonality of nucleophosmin/nucleoplasmin in two cyprinid fish.

Nucleophosmin/nucleoplasmin has been studied mostly in mammals and amphibians. To clarify the characteristics and function of nucleophosmin/nucleoplasmin in teleost fish, we cloned a full-length cDNA sequence from two cyprinid fish, Carassius auratus gibelio and Carassius auratus. Molecular characterization and multiple sequence alignments suggested that they are the homologs of nucleophosmin. RT-PCR and Western blot detected a specific expression in gonads, and immunofluorescence localization revealed their distribution in oogenic and spermatogenic cells. Furthermore, a sperm decondensation function was demonstrated by immunodepletion and in vitro sperm decondensation experiments. The data suggest that the cloned nucleophosmin should share expressional and functional characterization with nucleoplasmin and therefore provide novel evidence for a functional commonality of nucleophosmin and nucleoplasmin in fish.

[Study on the modified surface layers of the CIGS thin films by Raman spectra].

In the present paper, the properties of Cu(In(1-x) Ga(x)) Se2 (CIGS) thin film absorber materials for the solar cells obtained by selenization of the precursors with In-rich or CuGa-rich surface layers were studied by XRD, SEM and Raman spectra. The photovoltaic devices based on the absorbers were measured and analyzed by illuminated J-V curve subsequently. The performance of the device constructed by the absorbers obtained by selenization of the precursors with CuGa-rich surface layer was improved greatly compared to that with In-rich surface layer. Through Raman spectra measurement, it was found that the Raman peak of the A1 mode was shifted for the CuGa-rich one, which is verified that the band gap of the surface layers was elevated. Moreover the value of increased Ga contents within the surface region of films were calculated by the relation between the Raman shifts and the Ga contents. As a result, the devices based on the thin films with the elevated surface energy band by selenizing the precursors with the CuGa-rich surface layer improved further the V(oc) and FF by about 74 mV and 8% respectively compared to that of corresponding to the one with In-rich surface layers, so that the conversion efficiency of the photovoltaic devices based on these thin films with CuGa-rich surface layer was improved by up to 9.4%. Meanwhile Raman scattering spectroscopy has proven to be a very powerful and useful technique for the surface analysis of such thin film solar cell semiconducuor materials.

Comparative studies on in vitro sperm decondensation and pronucleus formation in egg extracts between gynogenetic and bisexual fish.

A cell-free system based upon the egg extracts from gynogenetic gibel carp (Carassius auratus gibelio) or bisexual red common carp (Cyprinus carpio red variety) was developed to investigate developmental behaviors of the demembranated sperm nuclei. Both red common carp and gibel carp sperm nuclei could decondense fully and form pronuclei in the red common carp egg extracts. Gibel carp sperm nuclei could also decondense fully and form pronuclei in the gibel carp egg extracts, but red common carp sperm nuclei could not decondense sufficiently in the same extracts. The significant differences of morphological changes were further confirmed by ultrastructural observation of transmission electron microscopy. The data further offer cytological evidence for gonochoristic reproduction in the gynogenetically reproducing gibel carp. In addition, the sperm nuclei in vitro decondensation is dependent on the pH in the extracts, and the decondensed efficiency is optimal at pH 7. However, no DNA replication was observed in the two kinds of egg extracts during the incubation period of the sperm nuclei. It is suggested that the egg extracts prepared from the gynogenetic gibel carp should be a valid in vitro system for studying molecular mechanism on gynogenesis and reproduction mode diversity in fish.