Eosin Research Articles

Iso-and Xeno-transplantation of kidney metanephroi preserved in histidine-tryptophan-ketoglutarate solution

Objective To evaluate the preservation effect of histidine-tryptophan-ketoglutarate (HTK) solution compared to Uinversity of Wisconsin Solution (UW) solution on organogenesis and function of embryonic metanephroi using an isograft or athymus animal model. Methods Whole metanephroi anlages from Lewis rats on embryonic day 16 (E16) immediately preserved in HTK or UW solution for 24, 48, 72 and 96 h at 0-4 ℃, were implanted in omenta of Lewis (Lewis→Lewis) rats or BALB/c (Lewis→BALB/c) nude mice. In vitro, Levels of lactate dehydrogenase (LDH; cell necrosis) and adenosine 5’-triphosphate (ATP) reserve of intracellular energy were measured for preserved E16 metanephros primordia. in vivo, developed metanephros primordia were observed for histopathological and electronic microscopy (EM) examination, or measured for graft survival rate and for clearance rate of inulin. Results Though release rate of LDH was lower in HTK at (6.5±1.1)%, (8.6±0.9)%, (11.3±1.3)%, (12.5±0.8)%, than in UW solution, yet had no significantdifference (F=0.752, P>0.05) preserved for 24, 48, 72 and 96 h. The ATP reserve (%) was significantly higher after ischemia (t=-2.725, P 0.05) up to 48 h and then 96 h in HTK and UW solution. Pre-implantation, the size and extent of tissue differentiation of E16 metanephroi preserved in HTK solution for 3 days were not so distinguishable from the size and differentiation of same chronological age such as E19 metanephroi. Four weeks post-implantation, E16 metanephroi had formed mature nephrons and collecting ducts. Hematoxylin and eosin (HE) and EM examination showed they had normal cellular and ultra structure. E16 metanephroi iso-or xeno-grafted after preservation in HTK and UW solution for 3 days had no significantly different values of graft survival rate at 100.0%, 100.0%, 73.3%, 0% (χ2=1.553, P>0.05) and 100.0%, 100.0%, 80.0%, 0% (χ2=0.653, P>0.05), and for clearance rate of inulin [(32.10±5.09) μl/(min·g), t=-0.264, P>0.05] after 8 weeks post-implantation. Conclusion Iso-or xenografted E16 metanephroi undergo growth and differentiation and exhibit excretory function after preservation in HTK solution for 3 days. Isograft and xenotransplant of metanephros anlage may be used as an effective animal model for comparing preservation effect of graft organ solutions. Key words: Metanephros; Homologous transplantation; Xenotransplantation; Histidine-tryptophan-ketoglutarate solution; Organogenesis

Effects of hypoxic-induced bone marrow mesenchymal stem cells on motor and cognitive function in rats with craniocerebral trauma

Objective To explore the effects of hypoxic-induced bone marrow mesenchymal stem cells (BMSCs) on motor and cognitive function in rats with craniocerebral trauma. Methods 18 adult Wistar rats were randomly divided into 3 groups, Group A, Group B, Group C and another 6 normal rats were taken as the normal control group. The Morris water maze test was performed on 21, 23 and 25 days after injection of BMSCs or dulbeeeo modified eagle medium (DMEM) solution, including the location-based navigation test and the spatial search test. Hematoxylin (HE) staining was performed on the hippocampus tissue after the experiment. Western blotting was used to detect the expression of apoptotic marker protein. Results The incubation period of rats in the four groups showed a decreasing trend with time (F=22.460, P 0.05). The space search experiment shows that the number of crossing in the group C and the swimming time in the platform quadrant were higher than those in the group A and B, but lower than the control group (F=198.670, P<0.01; F=4.615, P<0.05). The observation results of hematoxylin and eosin (HE) staining showed that there was a decrease in the number of nucleus parenchyma neurons in the hippocampal area in group A. There were significant differences in group B and C compared to group A. Westernblotting results indicated that group C had better anti-apoptotic effect than group B [(0.51±0.13) vs. (0.86±0.21), t=3.471, P<0.05]. PCR results indicated that group C had better anti-apoptotic effect than group B [(0.41±0.08) vs. (0.78±0.13), t=5.937, P<0.01]. Conclusion BMSCs induced by hypoxic conditions have better effects on motor and cognitive function in rats with craniocerebral trauma than normal oxygen induction. Key words: Bone marrow mesenchymal stem cells; Craniocerebral injury; Rat; Low oxygen

The classification and clinicopathological analysis of hepatocellular adenoma based on immunohistochemistry

Objective The subtypes of hepatocellular adenoma (HCA) were classified by immunohistochemical study, and the clinicopathological characteristics of each subtype were analyzed. Methods From December 2003 to March 2018, 31 cases with HCA were retrieved from the archive files of the Department of Pathology, Peking Union Medical College Hospital, including 16 male patients and 15 female patients. The age ranged from 16 to 63 years. Hematoxylin and eosin (HE) and immunohistochemical staining were performed with HCA samples. The subtypes were classified by immunohistochemical staining, and the clinicopathological characteristics of each subtype were analyzed. Results The HCA patients had no obvious and specific clinical symptoms, and most of them were diagnosed during the routine health checkup. All of the 31 patients were treated with surgery, and the complete resection was achieved in 26 cases. The adenomas were mainly in the right lobe of the liver (51.6%, 16/31), and the solitary adenoma accounted for 54.8% (17/31). 8 HCA were classified as H-HCA, 14 as I-HCA, 7 as β-HCA, and 2 as U-HCA by immunohistochemistry. H-HCA was characterized histologically by marked steatosis and lobulated contours, lacking L-FABP staining. I-HCA exhibited inflammatory infiltrate, telangiectasia, thickened arteries, more or less obvious ductular reaction, with the positive L-FABP/SAA/CRP staining. β-HCA all showed fibrous capsules, some of adenomas exhibited pseudoglandular structure and nodules in nodule. A strong homogeneous cytoplasmic overexpression of GS and nuclear β-catenin were observed in all β-HCA cases. The staining of SAA/CRP/GS was lacking in the U-HCA. Conclusions HCA is rare and difficult to diagnose in clinic. Final diagnosis relies on histological features, and immunohistochemical examinations need to be used for subtyping classification. Each of the four subtypes has characteristic pathological features. Key words: Adenoma; Pathology; Diagnosis; Immunohistochemistry; Classification

SnO2/Fe2O3/Ag Nanocomposite via Hydrothermal Approach: A Novel Highly Efficient Photodegradation of Eosin Yellow and Brilliant Green Dyes Under Visible Light Irradiation

The health hazards generated from textile and dye industries wastewater which are most carcinogenic and they poses a danger to human and aquatic biota. The present work investigates the degradation of dye pollutants such as Eosin yellow (EY) and Brilliant green (BG) under visible light irradiation using SnO2/Fe2O3/Ag nanocomposite. This nanocomposite was prepared by hydrothermal route and characterized by various instrument techniques like XRD, FTIR, FESEM, EDS, HRTEM, UV–Vis DRS, PL and UV–Vis spectrophotometer for determine its crystalline phase, morphology, bandgap and photocatalytic efficiency. The photocatalytic activity of nanocomposite was tested on both dyes EY and BG under visible light irradiation and the experimental results declared that this composite shown better photocatalytic activity in basic solution degraded 93% of BG and 95% of EY in 75 min and 60 min, respectively under the normalised conditions such as basic pH, 30 mg catalyst dose and 10 mg/L of dye solution concentration without any oxidants such as H2O2. This is due to lower bandgap energy of SnO2/Fe2O3/Ag nanocomposite (1.78 eV) and thus shows the red shift, making it active in the visible region, resulting in higher photocatalytic activity. This was proved by compare the photocatalytic efficiency with SnO2 and SnO2/Fe2O3 shown lesser photocatalytic degradation efficiency over EY and BG under visible light irradiation.

Effect of exogenous insulin on IRE1α-XBP1 signaling pathway in pancreatic tissues during cardiopulmonary bypass-caused insulin resistance in rabbits

Objective To evaluate the effect of exogenous insulin on inositol-requiring protein 1α (IRE1α)-X-box-binding protein 1 (XBP1) signaling pathways in pancreatic tissues during cardiopulmonary bypass (CPB)-caused insulin resistance in rabbits. Methods Forty healthy adult New Zealand white rabbits of both sexes, weighing 2.5-3.0 kg, were divided into 4 groups (n=10 each) using a random number table method: control group (group C), group CPB, insulin group (group I), and normal saline control group (group NS). CPB was established in group CPB.Insulin was intravenously infused in a dose of 1.2 ml/h from establishing CPB to 1 day after operation, and the infusion rate of insulin was regulated according to the blood glucose (maintaining at 7.2-8.3 mmol/L) in group I. CPB was established, and normal saline was intravenously infused from the beginning of operation to 1 day after operation in group NS.Before CPB (T1) and at 15, 30 and 60 min after aortic opening (T2-4), blood samples were collected from the left femoral artery, the plasma was separated, the blood glucose level was detected by oxidase method, the level of glucagon was detected by the radioimmunoassay method, and the insulin resistance index was calculated.Animals were sacrificed at T4, and pancreatic tissues were obtained for determination of the expression of IRE1α, XBP1, c-Jun N-terminal kinase(JNK)and caspase-12 protein and mRNA (by Western blot or fluorescent quantitative real-time polymerase chain reaction) and for examination of the pathological changes (by haematoxylin and eosin staining). Results Compared with group C, blood glucose and glucagon concentrations and insulin resistance index were significantly increased at T2-4, and the expression of IRE1α, XBP1, JNK and caspase-12 was up-regulated at T4 in CPB, I and NS groups (P 0.05). Conclusion The mechanism by which exogenous insulin reduces CPB-caused insulin resistance may be related to inhibiting IRE1α-XBP1 signaling pathways in pancreatic tissues of rabbits. Key words: Insulin; Cardiopulmonary bypass; Insulin resistance; Nositol requiring enzyme-1α; X-box-binding protein 1

Role of CaN/NFATc4 signaling pathway in ventilator-induced lung injury

Objective To evaluate the role of calcineurin (CaN)/nuclear factor of activated T cell cytoplasmic 4 protein (NFATc4) signaling pathway in inflammatory responses in lung tissues of rats with ventilator-induced lung injury (VILI). Methods Twenty-four clean-grade healthy male Wistar rats, aged 5-8 weeks, weighing 220-250 g, were divided into 3 groups (n=8 each) using a random number table method: control C (group C), VILI group and cyclosporine A plus VILI group (group CsA+ VILI). The animals were anesthetized with pentobarbital and tracheostomized. The rats were mechanically ventilated for 6 h with the tidal volume set at 40 ml/kg and respiratory rate at 40 breaths/min to establish the model of VILI.The rats kept spontaneous breathing in group C. CaN specific inhibitor cyclosporine A 10 mg/kg was intraperitoneally injected at 1 h before ventilation in group CsA+ VILI.Rats were sacrificed immediately after ventilation, lung tissues were obtained and stained with hematoxylin and eosin to evaluate lung injury, broncho-alveolar lavage fluid was collected for determination of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 concentrations by enzyme-linked immunosorbent assay, and the lungs were removed for determination of the wet to dry weight ratio (W/D ratio), expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) (by real-time polymerase chain reaction), and expression of calcineurin and NFATc4 in lung tissues (using Western blot). Results Compared with group C, the W/D ratio, lung injury scores and concentrations of IL-1β, IL-6 and TNF-α in BALF were significantly increased, and the expression of CaN, NFATc4, ICAM-1 mRNA and VCAM-1 mRNA was up-regulated in group VILI (P<0.05). Compared with group VILI, the W/D ratio, lung injury scores and concentrations of IL-1β, IL-6 and TNF-α in BALF were significantly decreased, and the expression of CaN, NFATc4, ICAM-1 mRNA and VCAM-1 mRNA was down-regulated in group CsA+ VILI (P<0.05). Conclusion CaN/NFATc4 signaling pathway mediates inflammatory responses in lung tissues of rats with VILI. Key words: Ventilator-induced lung injury; Calcineurin; NFATC transcription factors; Inflammation

Role of α7nAChR in postoperative cognitive dysfunction in aged rats with tibial fracture

Objective To evaluate the role of α7 nicotinic acetylcholine receptor (α7nAChR) in postoperative cognitive dysfunction in aged rats with tibial fracture. Methods One hundred and fifty clean-grade healthy male Sprague-Dawley rats, aged 18-22 months, weighing 440-580 g, were divided into 5 groups (n=30 each) using a random number table method: control group (group C), sham operation group (group S), tibial fracture group (group T), normal saline group (group N) and α7nAChR agonist PUN282987 group (group P). Group C received no treatment.Ten percent chloral hydrate 0.4 ml/100 g was injected intraperitoneally in group S. Group T underwent tibial fracture.PUN282987 2.4 mg/kg was intraperitoneally injected at 5 min before tibial fracture in group P. The equal volume of normal saline was given at 5 min before tibial fracture in group N. Morris water maze test was performed at day 7 after surgery.At days 1, 3 and 7 after surgery, the pathological changes of the hippocampal CA3 region were observed by haematoxylin and eosin staining, and the expression of α7nAChR, choline acetyltransferase (ChAT), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the hippocampal CA3 region was measured by Western blot. Results Compared with group C, the postoperative escape latency and swimming distance were significantly prolonged, and the expression of α7nAChR, ChAT, TNF-α and IL-1β was up-regulated at each time point after operation in T, N and P groups (P 0.05). Compared with group T, the postoperative escape latency and swimming distance were significantly shortened, and the expression of α7nAChR and ChAT was up-regulated and the expression of TNF-α and IL-1β was down-regulated at each time point after operation in group P (P 0.05), and the pathological changes of the hippocampal CA3 region were significantly attenuated in group P. Conclusion α7nAChR antagonism is involved in the development of postoperative cognitive dysfunction in aged rats with tibial fracture. Key words: Cholinergic agonists; Aged; Cognition disorders; Postoperative complications

Study on the relationship of transforming growth factor-β1 and a disintegrin and metalloproteinase with thrombospondin motif-4 in carotid atherosclerotic plaques

Objective To investigate the relationship between the expression level of transforming growth factor-β1 (TGF-β1) and a disintegrin and metalloproteinase with thrombospondin motif-4 (ADAMTS4) in human carotid atherosclerotic (AS) plaques. Methods Plaque tissues (n=46) were collected after carotid endarterectomy and divided into stable plaque group and unstable plaque group according to the histopathological features by hematoxylin and eosin (HE) staining. Immunohistochemistry (IHC) and real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) were used to detect the expression of ADAMTS4 and TGF-β1 in plaques, respectively. Umbilical arteries were served as the control group (n=10) and statistics analysis were conducted as needed. Results IHC results showed that both of ADAMTS4 and TGF-β1 were expressed in all plaques; RT-qPCR results showed that the expression level of ADAMTS4 mRNA was significantly higher in unstable plaque group (4.43±2.02) compared to the stable plaque group (3.16±1.57) and the controls (1.02±0.22, F=17.545, P<0.05); TGF-β1 mRNA in stable plaque group (4.68±1.42) was also higher than that in the unstable plaques (3.57±1.76) and the controls (1.14±0.31), and the difference was statistically significant between groups (F=24.676, P<0.05). Pearson correlation analysis showed that there existed a positive correlation between indexes (r=0.439, P<0.05). Conclusion ADAMTS4 and TGF-β1 were increased together during the process of AS and plaque evolution, which may be related to dose or cell type dependence. Key words: Carotid atherosclerosis; Unstable plaques; Transforming growth factor-β1; A disintegrin and metalloproteinase with thrombospondin motif-4

Effect of acid sensitive ion channel 3 on hypoxia on microenvironment of nucleus pulposus cells and its mechanism

Objective To investigate the effect of hypoxia on acid sensitive ion channel 3 (ASIC3) in nucleus pulposus microenvironment and its mechanism. Methods Primary nucleus pulpocytes were isolated and identified by immunohistochemistry, hematoxylin and eosin (HE) and toluidine blue. Immunohistochemistry, HE, toluidine blue and real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) were used to explore the appropriate hypoxic concentration, apoptosis and FQ-PCR to explore the appropriate action time. The interference and overexpression of ASIC3 vector were constructed, and their efficiency was verified by FQ-PCR and Western blotting. The experiment was divided into the normal group (Control). Hypoxic treatment group (2%O2); shRNA-NC+ 2%O2); shRNA-ASIC3+ 2%O2; Expression of no-load + hypoxic treatment (Vector+ 2%O2); ASIC3 expression + hypoxic treatment group (ASIC3+ 2%O2). The proliferation was detected by cell counting kit-8 (CCK-8), the cell cycle and apoptosis were detected by flow cytometry, the formation of autophagosomes in each group was detected by transmission electron microscopy, the expression of hypoxia inducible factor (HIF)-1 and ASIC3 was detected by immunofluorescence, the expression of HIF-1, light chain 3 (LC3), extracellular signal-regulated kinase 1/2 (ERK1/2) and mitogen-activated protein kinase (MAPK) and phosphorylation were detected by FQ-PCR and Western blotting, and the effect of ASIC3 on HIF-1 and ERK1/2 was detected by immunoco-precipitation. Results Nucleus pulposus cells were successfully isolated with appropriate hypoxic concentration of 2% and appropriate action time of 48 h. Both interference and overexpression vector construction were successful and effective. Compared with the control group, cell proliferation was inhibited in the hypoxic group and ASIC3 expression + hypoxic treatment group, apoptosis was increased, cell cycle arrest was in G1 phase, calcium ion concentration was increased, and HIF-1, LC3, p-ERK1/2 and p-mapk expression were increased, the difference was statistically significant (P<0.05). Conclusion Interference with ASIC3 can inhibit the activation of mitogen-activated protein kinase (MAPK) signal to improve the microenvironment of hypoxic nucleus pulposus cells, which may be one of the pathogenesis of discogenic low back pain. Key words: Nucleus pulposus cells; Discogenic low back pain; Acid-sensing ion channel

Kinetic and Equilibrium Modeling of Anionic Dye Adsorption on Polyaniline Emeraldine Salt: Batch and Fixed Bed Column Studies

In this work, removal of anionic dye Eosin Yellow (EY) by polyaniline emeraldine salt (PANI-ES) from the aqueous medium was studied by using both batch and fixed-bed column mode. Before and after the dye adsorption, the adsorbents were characterized by UV-Vis, FTIR, and XRD analysis. The morphology of the adsorbent was also studied by scanning electron microscopy (SEM). The batch adsorption study was performed by varying the initial concentration of the dye and pH. The experimental results obtained were fitted to the three isotherm models namely Langmuir, Freundlich, and Temkin and the Langmuir isotherm was found to be the best fitting model with the adsorption capacity 335 mg/g. The adsorption process followed pseudo second order kinetics. The thermodynamic study was also performed and thermodynamic parameters ΔH°, ΔS°, and ΔG° were calculated. The adsorption experiments were also carried out in packed columns by varying some important factors like flow rate, bed height, pH, and the initial dye concentration. The experimental results obtained were fitted using Bohart-Adams, Yoon-Nelson, and Thomas models. Among the models studied, the highest adsorption capacity, 79.0 mg/g was calculated from Thomas model at pH 4 with 375 mg/l initial concentration, flow rate 0.3 ml/min, and bed height 1.4 cm.

Multifunctional Nanohybrid of Palladium Nanoparticles Encapsulated by Carbon‐Dots for Exploiting Synergetic Applications

AbstractPalladium nanoparticles encapsulated in the carbon dots (Pd/C‐dots) are demonstrated to play a role of multifunctional nanohybrid in the synergetic applications of sensor and catalysis. The photochemical method is applied to synthesize Pd/C‐dots in which Pd nanoparticles (NPs) are dispersedly encapsulated by C‐dots layer. The nanohybrid can function as a fluorescent sensor and reductive catalyst, due to the inherent properties of C‐dots and Pd NPs, respectively. The Pd/C‐dots exhibit a highly selective and sensitive detection toward the nickel (Ni2+) ion with a detection limit of 7.26 × 10−9 m. Moreover, the Ni2+ is detected in MCF‐7 live cells signifying the applicability of nanohybrid as a promising sensor. On the other hand, the Pd/C‐dots show an excellent catalytic performance in the reduction of 4‐nitrophenol and eosin yellow. A plausible mechanism for sensing and catalysis behavior is proposed. The sensor system is designed on the basis of the fluorescence turn‐on when Ni2+ interacts with functional groups of the C‐dots layer. The activities of catalytic reduction are mainly governed by the Pd NPs and further enhanced when the C‐dots layer is incorporated. The Pd/C‐dots can serve as a new paradigm for opening a potential trend in the design of multifunctional materials to diverse applications.

Effect of mild hypothermia combined with hydrogen-rich saline on cerebral injury after cardiac arrest and resuscitation in rats

Objective To evaluate the effect of mild hypothermia combined with hydrogen-rich saline on cerebral injury after cardiac arrest and resuscitation in rats. Methods Healthy male Sprague-Dawley rats, aged 7-8 weeks, weighing 280-320 g, were divided into 5 groups (n=33 each) using a random number table method: sham operation group (group S), cardiac arrest and resuscitation group(group CA-R), hydrogen-rich saline group (group H2), mild hypothermia group (group MH), and mild hypothermia plus hydrogen-rich saline group (group MH+ H2). Cardiac arrest was induced with transoesophageal cardiac pacing followed by cardiopulmonary resuscitation to establish the cerebral injury model.Hydrogen-rich saline 5 ml/kg was intraperitoneally injected immediately after return of spontaneous circulation (ROSC) in H2 and MH+ H2 groups, while the equal volume of normal saline was given instead in the other groups.The body temperature of rats was cooled down to 32-34℃ within 15 min starting from the time point immediately after ROSC and maintained for 4 h in MH and MH+ H2 groups.Fifteen rats were selected at 24 h after ROSC to assess the neurological function score (NDS). Eighteen rats in each group were sacrificed at 24 h after ROSC, and brains were removed for microscopic examination of the pathological changes in hippocampal CA1 region after hematoxylin and eosin staining and for determination of pyramidal cell count and expression of glucose-regulated protein 78 (GRP78), C/EBP-homologous protein (CHOP), caspase-12, caspase-3, Bcl-2 and Bax in hippocampal CA1 region (by Western blot). Results Compared with group S, the NDS was significantly decreased, the pyramidal cell count was reduced, the expression of GRP78, CHOP, caspase-12, caspase-3 and Bax was up-regulated, and the expression of Bcl-2 was down-regulated in the other four groups (P 0.05). Conclusion Combination of mild hypothermia and hydrogen-rich saline offers enhanced efficacy in reducing cerebral injury after cardiac arrest and resuscitation over mild hypothermia or hydrogen-rich saline alone in rats. Key words: Hypothermia, induced; Hydrogen; Heart arrest; Cardiopulmonary resuscitation; Reperfusion injury; Brain

The mechanism of CD8+ CD45RClow regulatory T cell in rat liver transplantation nature tolerance model

Objective To explore the mechanism of CD8+ CD45RClow regulatory T cells and its related cytokines function during the tolerance process of orthotopic liver transplantation in rats. Methods The male Brown Norway rats were selected as the donor, the Lewis rats were the recipients, the allogenic orthotopic liver transplantation model was established by the double cuff method. Experiment animals were divided into early transplantation group (10 rats), tolerance group (10 rats), sham group. Liver function were detected. The tolerance recipients were detected by hematoxylin and eosin (HE) staining. The Luminex used to detect of cytokine and the flow cytometry were used to analyze the changes of subsets of lymphocytes in the peripheral blood/graft and draining lymph node. Results 70% of recipients in operation group could be induced natural immune tolerance. The levels of aspartate aminotransferase (AST) in tolerance group were [(456±54) U/ml] vs. those of sham group [(460±151) U/ml], (P>0.05); the levels of alanine aminotransferase (ALT) in tolerance group were [(122±35) U/ml] vs. those of sham group [(121±46) U/ml, P>0.05]. The expression of CD8+ CD45RClow T cells in tolerance group were increased than in short term group; in grafts it was up regulated in tolerance group than in sham group, and it was no significance between sham group and tolerance group in lymph node around graft. The expression of pDC in short term group are significantly increased compared with sham group. The interferon (IFN)-γ was increased in short term group, return to normal level in tolerance group. interleukin (IL)-10 in grafts milieu were dramatically increased, keep in high level in tolerance group. Conclusion In the tolerance recipients of rat orthotopic liver transplantation model, CD8+ CD45RClow regulatory T cells were significantly increased and aggregated in the peripheral blood and liver. The inhibitory cytokines IL-10 was important to maintain the long-term graft tolerance. Key words: Regulatory T cell; Liver transplantation; Immune tolerance; Interleukin-10; Dendritic cell

Graphene oxide clay nanocomposite as an efficient photo-catalyst for degradation of cationic dye

Development of nanocomposite for remediation of environmental pollutants like dye is essential in present day world. In this work, we successfully prepare graphene oxide–clay nanocomposite by different methods. The nanocomposite was characterized using various characterization techniques viz. Uv–visible spectroscopy, Fourier Transformed Infra-red spectroscopy, Dynamic light scattering, X-ray diffraction, and scanning electron microscopy. The graphene oxide-clay nanocomposite was very effective catalyst against photodegradation of exclusively cationic dye like methylene blue. But the prepared nanocomposite was not found to be effective catalyst against degradation of other anionic dyes like methyl orange and Eosin yellow. The graphene-oxide clay nanocomposite was found to be an efficient photocatalyst in degradation of methylene blue when compared with only clay or graphene oxide. It is also demonstrated in the work that the nanocomposite catalyst can be reused several times without effecting its efficiency, the percentage degradation was 85%, 75% and 75% for first, second and third cycle respectively.The mechanistic insight of clay-graphene oxide as a catalyst for only cationic dye is also discussed in the paper.

Immunomodulation combined with bone marrow mesenchymal stem cells improves sepsis-induced acute lung injury

Objective To explore the appropriate treatment opportunity and the possible mechanism of ulinastatin combined with bone marrow mesenchymal stem cells (BMSCs) improving sepsis-induced acute lung injury. Methods The cecum end catheter continuous drainage (CASP) was used to establish septic model in 40 male SD rats weighing 180-220 g. The animals were randomly divided into 5 groups (n=8 each group): control (A), septic group (B), ulinastatin combined with BMSCs treatment group at the same time (group C), BMSCs intervention 18 h after administration of ulinastatin (group D), BMSCs intervention 66 h after administration of ulinastatin (group E). The hematoxylin and eosin (HE) staining, immunohistochemistry and Western blotting were applied to examine the lung tissue pathologic changes, and alveolar surface activated protein C and E-calcium expression of mucin respectively. The survival curve was used to expore the survival rate. Results The survival rate among groups had no statistically significant difference (P>0.05). Pathological scores of rat lung tissues in groups A-E were 2.92±0.43, 14.22±1.92, 8.67±1.21, 7.50±1.76 and 10.80±1.47 respectively. The pathological injury of lung tissue in the intervention groups was milder than in group B (P 0.05). Conclusion After immunomodulation, the lung injury caused by sepsis in rats was the mildest within 18 h. The possible mechanism is contributed to the increases in the synthesis of alveolar surface active protein C, and inhibition of the loss of e-cadherin in lung tissue, which can improve the microenvironment of the lung tissue, and delay the lung fibrotic process of sepsis-induced acute lung injury. Key words: Sepsis; Acute lung injury; Pulmonary surfactant protein C; E-Cadherin

Effects of varicocele on 4-aminobutyrate aminotransferase expression and its potential role in surgically varicocele-induced rat testes

Objective To investigate the expression of 4-aminotransferase (ABAT) and its effect on spermatogenesis in experimental varicocele rat model. Methods A total of 36 specific pathogen free (SPF) male SD rats were randomly divided into varicocele group and sham operation group, with 18 rats in each group. The experimental rat varicocele was induced by partially occluding left renal vein and completely ligating the branches of the left spermatic vein (LSV). After 4 weeks, the using hematoxylin and eosin (HE) staining, Masson and glycogen staining were used to evaluate the two groups of kidney and testis histology change, evaluation of varicocele rat model. After the model was constructed, the localization of ABAT protein in rat testis was further analyzed by immunofluorescence, and the expression of ABAT protein in varicocele was detected by Western blotting, and the possible role was analyzed. Results 4, 6, 8 weeks after the operation, the varicocele group in the kidney tissue pathological changes were not found, and the left spermatic vein diameter was greater than 1 mm [(1.52±0.23) mm vs. (0.26±0.03) mm], the weight of the left testis [(1.61±0.08) g vs. (1.93±0.11) g] and the percentage of seminiferous tubules (58% vs. 18%) were lower than the control group, which indicates that the successful construction of varicocele animal model. Immunofluorescence analysis showed that ABAT protein was mainly expressed in the cytoplasm of spermatogenic cells. Compared with the control group, with the prolonged duration of varicocele, Western blotting showed that the expression of ABAT protein was down regulated in the varicocele group, which is positive with the degree of spermatogenesis damage. Conclusion In experimental rat varicocele model, varicocele affects the expression of ABAT protein, which may be related to varicocele damage spermatogenesis. Key words: 4-transaminase; Varicocele; Rat testis

CD24 regulates Ly6Chi macrophages and BDL-induced liver fibrosis

Objective To investigate the regulatory effects of CD24 on Ly6Chi macrophages in liver and its influences on bile duct ligation (BDL)-induced hepatic fibrosis in mice. Methods Liver fibrosis was induced in wild-type (WT) and CD24-/- mice by surgical ligation of the biliary duct. Levels of alanine amino transferase (ALT) in serum were detected and liver sections were stained with haematoxylin and eosin (H&E) staining to assess the severity of liver injury. Sirius Red staining was used to observe the degree of liver fibrosis. Real-time PCR was performed for detecting the expression of hepatic fibrosis-related markers and TGF-β1 at mRNA level. The percentage of macrophages and the number of TGF-β1-producing macrophages were measured by flow cytometry. Results BDL-induced liver fibrosis was exacerbated in CD24-/- mice than in WT mice as demonstrated by more serious hyperplasia in bile duct, more inflammatory infiltration at the portal area and higher levels of ALT in serum. Results of Sirius red staining also showed that the liver fibrosis was more severe in CD24-/- mice than in WT mice. Moreover, α-SMA and collagen typeⅠ alpha 1 (Col1a1) were significantly upregulated in CD24-/- mice. Flow cytometry analysis revealed that CD24 was highly expressed by hepatic macrophages in BDL-induced WT mice, and the percentages of hepatic macrophages were significantly elevated in CD24-/- mice compared with those in WT mice. Further analysis revealed that the percentages of Ly6Chi hepatic macrophages in CD24-/- mice were higher than those in WT mice, but there was no significant difference in the percentages of Ly6Clo macrophages. The expression of TGF-β1 at mRNA level was increased in CD24-/- mice as compared with that in WT mice after BDL. Moreover, intracellular staining showed that Ly6Chi hepatic macrophages in CD24-/- mice secreted more TGF-β1 than the macrophages in WT mice. Conclusions CD24 might attenuate the BDL-induced liver fibrosis in mice via regulating the percentage of hepatic Ly6Chi macrophages and the secretion of TGF-β1. Key words: CD24; Bile duct ligation (BDL); Liver fibrosis; Ly6Chi hepatic macrophage; TGF-β1

Therapeutic effects of baicalin on lupus nephritis in a lupus-prone mouse model and its regulatory role in follicular helper T cell differentiation

Objective To evaluate the therapeutic effect of baicalin on lupus nephritis in a lupus-prone mouse model, and to investigate its regulatory role in the differentiation of follicular helper T (Tfh) cells. Methods Eight 12-week-old female MRL/lpr lupus-prone mice were randomly and equally divided into two groups by a random number table i.e., baicalin group and control group intraperitoneally injected with 200 mg/kg baicalin sodium and chloride physiological solution, respectively, once every day for 4 weeks. After the end of treatment, Coomassie brilliant blue staining was performed to detect the level of 24-hour urine protein. Then, the mice were sacrificed, and the spleens were resected and weighed. Mononuclear cells were isolated from these spleens, and flow cytometry was conducted to determine the proportion of Tfh cells. Additionally, the kidneys were resected and subjected to hematoxylin and eosin (HE) staining for the evaluation of kidney impairment. Moreover, some other mononuclear cells were isolated from the spleens of the lupus-prone mice in the control group, and magnetic activated cell sorting (MACS) was performed to isolate naive CD4+ T cells, which were divided into 3 groups: blank control group receiving no treatment, induction group treated with 10 μg/L anti-interleukin (IL) -21 and anti-IL-6 antibodies and 3 μg/L anti-CD3 and anti-CD28 antibodies for 5 days, and intervention group additionally treated with 40 μmol/L baicalin for 5 days besides the above treatment. Then, 50 μg/L phorbol ester, 750 μg/L ionomycin and 20 mg/L brefeldin A were used to stimulate some cultured naive CD4+ T cells in the above groups. Flow cytometry was conducted to determine the proportion of CD4+CXCR5+PD-1+ cells and CD4+IL-21+ cells. Statistical analysis was carried out with SPSS20.0 software by using one-way analysis of variance (ANOVA) and student t test for the comparison of quantitative data between groups. Results The baicalin treatment could effectively improve the kidney impairment in the lupus-prone mice. Compared with the control group, the baicalin group showed significantly decreased 24-hour urine protein level ([1 416 ± 171] vs. [2 623 ± 278] μg/24 h, P = 0.022) , and significantly decreased proportion of Tfh cells in the spleen (12.6% ± 2.3% vs. 40.2% ± 1.1%, P = 0.005) . In vitro baicalin could further inhibit the differentiation of Tfh cells. Compared with the induction group, the intervention group showed significantly decreased proportion of CD4+CXCR5+PD-1+ Tfh cells (13.3% ± 0.8% vs. 17.6% ± 0.9%, P = 0.04) and CD4+IL-21+ cells (1.0% ± 0.4% vs. 2.7% ± 0.2%, P < 0.01) . Conclusion Baicalin can effectively ameliorate lupus nephritis, which may be associated with the inhibition of Tfh cell differentiation. Key words: Lupus erythematosus, systemic; Lupus nephritis; Baicalin; Disease models, animal; Follicular helper T cells; Interleukin-21

Experimental and Computational Design of Highly Active Ce–ZrO2–GO Photocatalyst for Eosin Yellow Dye Degradation: The Role of Interface and Ce3+ Ion

As future photocatalyst require high activity and structural stability, the demand for high-performance photocatalyst that increases photocatalytic conversion efficiency is of significant interest. In this study, Ce–ZrO2–GO nanocomposite was fabricated by a co-precipitation technique and used for the photocatalytic degradation of eosin yellow (EY) dye under visible light irradiation. A hybrid DFT calculation was employed to analyse the electronic properties and the underlying mechanism of the observed photoactivity. The as-prepared Ce–ZrO2–GO (0.3% Ce) nanocomposite showed enhanced photodegradation of EY dye, which was about 15.98-fold higher than pure ZrO2. The enhanced performance of Ce–ZrO2–GO nanocomposite for EY dye degradation is due to the synergistic effect between the GO sheets and Ce–ZrO2. From the computational study, Ce ion could act as an electron mediator to capture the photoinduced electrons from ZrO2 to restrain the recombination rate of charge carriers. Testing with different scavengers showed that hydroxyl radicals play a major contribution to the degradation of EY dye with a pseudo first-order reaction kinetics. Total Organic Carbon analysis showed 76% mineralisation of EY dye over Ce–ZrO2–GO (0.3% Ce) nanocomposite, which is photostable and its efficiency remained almost the same after five cycles without any photocorrosion. The present study could offer an alternative approach to develop highly efficient photocatalysts for organic pollutant degradation using metal ion as a mediator to enhance charge migration, activity and structural stability of the nanocomposite. A series of Ce–ZrO2–GO nanocomposite were employed as a photocatalytic to degrade eosin yellow dye from wastewater. The present study offers an alternative approach to develop highly efficient photocatalysts for organic pollutant degradation using Ce ion as a mediator to enhance charge migration, activity and structural stability of the nanocomposite.

Inhibitory effect of bromfenac sodium hydrate ophthalmic solution on alkali burn induced-corneal neovascularization in rats

Objective To investigate the inhibitory effect of bromfenac sodium hydrate ophthalmic solution on corneal neovascularization (CNV) induced by alkali burn. Methods A total of 192 specific pathogen free (SPF) degree adult male Sprague-Dawley (SD) rats were used in this study.One hundred and seventy-two rats were chosen to establish CNV model with alkali burn in the right eyes.Following alkali burn, rats were randomly divided into CNV group, model control group, bromfenac sodium group and fluorometholone group, with 43 rats (43 eyes) in each group.Another 20 rats (40 eyes) served as normal control group.One day after modeling, the model control group, bromfenac sodium group and fluorometholone group received phosphate buffer saline (PBS), bromfenac sodium hydrate ophthalmic solution and 0.1% fluorometholone eye drops, respectively.The state of cornea and anterior chamber and the growth of CNV of rats in each group were observed by slit-lamp microscope every day after modeling.At 1, 3, 7, 14, 21 and 28 days after modeling, the anterior segment photos of the experimental eyes were captured, and the percent of cornea areas covered by CNV was calculated.At 7, 14 and 28 days after modeling, the eye tissue sections were stained with hematoxylin and eosin staining and immunohistochemistry staining to evaluate the expressions of CD45 and VEGF-A.Real-time quantitative PCR and enzyme-linked immunosorbent assay (ELISA) were used to detect the expression of COX-2 and VEGF mRNA and protein level.The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology(ARVO). Results Each model group showed corneal edema and opacification 1 day after modeling.The corneal edema was aggravated 7 days after modeling.On the 14th day after modeling, the degree of corneal opacity and edema decreased gradually.On the 28th day after modeling, leucoma was observed in CNV group and model control group, and nebula was observed in bromfenac sodium group and fluorometholone group.At 7, 14, 21 and 28 days after modeling, the percentages of CNV areas in bromfenac sodium group and fluorometholone group were significantly lower than those in CNV group and model control group (all at P 0.05). On the 7th day after modeling, the thinning of corneal epithelial layer, edema and arrangement disorder of stroma layer were observed, and the expression of VEGF-A was positive in all model groups; a small amount of CD45 positive inflammatory cell infiltrations were observed in CNV group and model control group.On the 14th and 28th day after modeling, CNV was seen in the center of cornea in CNV group and model control group; the epithelial keratosis and reduction of corneal edema were seen in each group, and no inflammatory cell infiltration was observed in each group.On the 7th day after modeling, the expressions of COX-2 and VEGF mRNA in CNV group and model control group were significantly higher than those in normal control group, bromfenac sodium group and fluorometholone group (all at P<0.05), the expressions of COX-2 and VEGF protein in bromfenac sodium group were significantly lower than those in CNV group (all at P<0.05). The corneal perforation rate in model control group and bromfenac sodium group was 10% (1 case in 10 rats). The corneal perforation rate in fluorometholone group was 30% (3 cases in 10 rats). In each model group, 10% to 30% rats had hyphema. Conclusions Bromfenac sodium hydrate ophthalmic solution can inhibit the formation and growth of CNV after alkali burn in rats.This effect may be mediated by regulating COX-2 expression, reducing inflammation and inhibiting VEGF production. Key words: Corneal neovascularization; Alkali burn; Bromfenac sodium; Cyclooxygenase-2