Objective To compare the expression of renal transforming growth factor-?_1(TGF-?_1),its receptors type Ⅰ and Ⅱ(T?RⅠ and T?R Ⅱ) in two rat models of chronic nephropathy induced by CsA and FK506 respectively.Methods Rat models of chronic CsA-/FK506-induced nephropathy were established by administering sandimun Neoral and Prograf separately.Then their kidneys were dissected and the protein expression of TGF-?_1,T?RⅠand T?RⅡ,the mRNA levels of T?RⅠ and T?RⅡ was detected by using immunohistochemistry(IHC) and in situ hybridization(ISH) respectively.Results The IHC revealed that the integrated optical densities(IODs) of renal TGF-?_1,T?RⅠ and T?RⅡ were(605.24)?(140.24),(876.28)?(208.73) and(981.59)?(258.65) respectively in CsA-treated rats,(488.37)?(101.76),(586.63)?(131.21) and(543.34)?(105.26) respectively in FK506-treated ones.The ISH indicated that the IODs of renal T?RⅠ and T?RⅡ mRNA were(905.08)?(158.75) and(1090.92)?(242.73) respectively in CsA-treated rats,(661.37)?(205.65) and(716.27)?(195.55) respectively in FK506-treated ones.The difference of the above-mentioned five factors between the two groups was significant(P

Adolescent ; Adult ; Astrocytoma ; diagnosis ; surgery ; Brain Neoplasms ; diagnosis ; surgery ; Child ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Microsurgery ; methods ; Middle Aged ; Thalamic Diseases ; diagnosis ; surgery ; Thalamus ; surgery ; Treatment Outcome

AIM:To be one of the primary cause injury to multiple sites of ocular of glaucoma which affects over 70 million people worldwide. We applied data mining techniques, linear and the matrix operations, efficiently calculated the network and estimated the possible function of the“node” genes of the retina and optic of glaucoma, in order to provide new thought and method on the pathogenesis of glaucoma.
METHODS: The data in this study is from Gene Expression Omnibus ( GEO ) which belong to Nation Center for Biotechnology Information ( NCBI) , the quality of the raw data CEL files was processed and analyzed by the Expression software which belong to Affymetrix Inc. , Santa Clare, CA, USA. Significant analysis method ( SAM) which base on the T test was used to identified the significant genes. Based on GRNInfer and Gvedit soft we set up gene networks of optic and retina of mice and further more enriched analysis which based on DAVID and MAS3. 0 online software were processed.
RESULTS:The analysis between the group of the optic nerve heads and retinas in different stage of glaucoma showed that the amount of significant different expressed genes in the optic never head group increased significantly comparing with the group of retina in the early stage of glaucoma, the analysis of the genes network construction show that:the node genes of optic nerve heads included Unc13c、Kif5a、TRPM1、PANX; and the node genes of retina include POU4F1, NEFL, BC03870, CALB2. Metabolic pathways enrichment analysis which based on MAS3. 0 online platform show that there was mainly the amyotrophic lateral sclerosis, tyrosine metabolism, melanogenesis, Nitrogen metabolism, Gap junction, Leukocyte transendothelial migration metabolism pathway enriched out in optic nerve head; and there was mainly amyotrophic lateral sclerosis, neurodegenerative disorders, prostate cancer, leukocyte transendothelial migration metabolism pathway enriched out in retina.
CONCLUSION:By understanding bioinformatics result, it seems optic were more sensitive than the retina to high intraocular pressure, and weather high expression of TYrp1 gene can be as a sensitive diagnostic item require more evidence back up. Functional enrich analysis of node gene showed that cytoskeleton reconstructed, molecular motor and nutrients transport function improve in optic; and in retina, the most prominent finding in retina was enrichment function modules were focus on regeneration, repairing and differentiation of cells, which remind that we should reinforce research on reparation of retina of primary glaucoma. Metabolic pathways enrichment analysis show that inflammatory response plays prominent place in optic and retina of primary glaucoma, because of the optic narrow and crowed anatomic shape, nutrient metabolism and substances transfer enrichment modules play an important role in optics of primary glaucoma.

Objective To investigate the relationship between the dynamic changes of interleukin-1β (IL-1β) levels and severity and complications of patients with multiple trauma at the early stage.Methods Among 97 patients with multiple trauma in Emergency Department of Shanghai Jiao Tong University Affiliated Sixth People's Hospital between August 2015 and May 2016,12 patients were excluded as follows,(1) with burns or chemical injuries;(2) pregnancy or menopausal women;(3) had bacterial infection a week ago;(4) with chronic diseases.The other 85 patients with multiple traumas were classified into three categories according to the injury severity score (ISS).That is,the slight group (22 cases,9≤ ISS ＜ 15),moderate group (35 cases,15 ≤ ISS ＜ 25) and severe group (28 cases,ISS ≥ 25).Their venous blood samples were collected at 6,12,24,48 and 72 hours after trauma respectively,and the serum IL-1 β levels were measured using a specific immunoluminometric assays.The basal conditions including age,the hospitalization days and so on among these three groups were compared via ANOVA.The mean IL-1 β levels at above time intervals among three groups were compared.Finally,the relationship between the peak concentration of IL-1β and injury severity and complications was analyzed by multiple Logistic regression.Results (1) As the increasing severity of trauma,the patients with longer days of hospitalization and higher rate of multiple organ dysfunction syndrome (MODS) (P ＜ 0.05).(2) The levels of IL-1 β in the moderate and severe groups were remarkably higher than those in the slight group (P ＜ 0.02).(3) The IL-1β levels in each group peaked at 6 hours after trauma and began to decline.(4) Multivariate logistic analysis showed that peak concentration of IL-1 β was still an independent predictor for injury severity (moderate group:odds ratio,1.21;95％ confidence interval:1.05-1.39,P =0.007;severe group:odds ratio,1.20;95％ confidence interval:1.03-1.40,P =0.019) and sepsis (odds ratio,1.28;95％ confidence interval:1.10-1.50,P =0.001),but had no significant association with MODS and trauma mortality even after controlling other risk factors.Conclusions The serum IL-1β at 6 hours after injury could be used as an early effective indicator to evaluate the injury severity and infectionrelated complications in patients with multiple trauma.

OBJECTIVETo investigate the changing laws of serum high mobility group box 1 protein (HMGB1) in septic rats and intervention effect of Xuebijing on it.

METHODSLipopolysaccharide (LPS) (5 mg/kg BW) was intravenously injected into the tail vein of healthy male Wistar rats to prepare the sepsis rat model. In Experiment 1: 50 Wistar rats were randomly divided into three groups, i.e., the normal group (A, n=10); the LPS model group (B, n=10), the LPS +Xuebijing treatment group (C, n=30). Rats in the C group were further divided into three subgroups, i.e., 2 h before LPS injection (group C1), 2 h after LPS injection (group C2), and 8 h after LPS injection (group C3), 10 in each group. Blood samples were collected from the caudal vein to detect serum HMGB1 levels by Western blot at 4, 12, 24, 48, and 72 h after LPS injection. Experiment 2: 30 Wistar rats were equally divided into the LPS model group (D) and the LPS + Xuebijing treatment group (E), 15 in each group. They were treated as rats in the B group and the C1 group respectively. Five rats were sacrificed at 12, 24, and 48 h after LPS injection in the two groups. Blood as well as the tissue samples were harvested to measure such indices as ALT, AST, Cr, and BUN, as well as pathological changes of liver, lung, and kidney.

RESULTS(1) Compared with the A group, serum HMGB1 levels were higher at various time points in the B group (P < 0.05). Compared with the B group, serum HMGB1 levels at 12,24,48, and 72 h decreased in the C1, C2, and C3 groups. Besides, the decrease was more obvious at 24 h and 48 h.The decrement in the C3 group was less than that in the C1 and C2 groups (P < 0.05). (2) In the D group, ALT, AST, Cr, and BUN were significantly higher than those in the A group and reached the peak at 24 h (P < 0.05). Compared with the E group, AST, Cr, and BUN at 24 and 48 h, and ALT at each time point decreased significantly in the E group (P < 0.05). (3)The results of pathological section of liver, lung, and kidney showed local congestion and hemorrhage, cell edema/necrosis/degeneration, infiltration of inflammatory cells, damage of characteristic structures and so on; particularly serious lesion occurred at 24 and 48 h in the D group. The microscopic lesion was obviously alleviated in the E group than in the D group at corresponding time points.

CONCLUSIONSThe serum HMGB1 levels increased in septic rats, with late occurrence of peak value and longer duration of the high value. HMGB1 played an important role in excessive inflammatory response and multiple organ dysfunction. Xuebijing could reduce the serum levels of HMGB1, improve biochemical parameters, and attenuate severe inflammatory response of liver, lung, and kidney tissues in septic rats. Besides, the earlier use, the better effect obtained.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; HMGB1 Protein ; blood ; Male ; Rats ; Rats, Wistar ; Sepsis ; blood ; drug therapy

BACKGROUNDSleep/wake disturbances in patients with amyotrophic lateral sclerosis (ALS) are well-documented, however, no animal or mechanistic studies on these disturbances exist. Orexin is a crucial neurotransmitter in promoting wakefulness in sleep/wake regulation, and may play an important role in sleep disturbances in ALS. In this study, we used SOD1-G93A transgenic mice as an ALS mouse model to investigate the sleep/wake disturbances and their possible mechanisms in ALS.

METHODSElectroencephalogram/electromyogram recordings were performed in SOD1-G93A transgenic mice and their littermate control mice at the ages of 90 and 120 days, and the samples obtained from these groups were subjected to quantitative reverse transcriptase-polymerase chain reaction, western blotting, and enzyme-linked immunosorbent assay.

RESULTSFor the first time in SOD1-G93A transgenic mice, we observed significantly increased wakefulness, reduced sleep time, and up-regulated orexins (prepro-orexin, orexin A and B) at both 90 and 120 days. Correlation analysis confirmed moderate to high correlations between sleep/wake time (total sleep time, wakefulness time, rapid eye movement [REM] sleep time, non-REM sleep time, and deep sleep time) and increase in orexins (prepro-orexin, orexin A and B).

CONCLUSIONSleep/wake disturbances occur before disease onset in this ALS mouse model. Increased orexins may promote wakefulness and result in these disturbances before and after disease onset, thus making them potential therapeutic targets for amelioration of sleep disturbances in ALS. Further studies are required to elucidate the underlying mechanisms in the future.

Amyotrophic Lateral Sclerosis ; genetics ; metabolism ; Animals ; Female ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Mice ; Mice, Transgenic ; Neuropeptides ; genetics ; metabolism ; Orexins ; Reverse Transcriptase Polymerase Chain Reaction ; Sleep ; physiology ; Superoxide Dismutase ; genetics ; metabolism ; Superoxide Dismutase-1 ; Wakefulness ; physiology

The total RNA was extracted from porcine ovary. Porcine Follistatin cDNA was cloned by RT-PCR. Complete porcine follistatin cDNA coding sequences are presented including 1038 bp of open reading frame. The purified porcine follistatin cDNA was inserted into pGEX-4T-3 vector to construct the prokaryotic fusion protein expression vector. The recombinant expression plasmid was transformed into BL21 (DE3) and expression was induced by IPTG. Protein products were detected by SDS-PAGE and confirmed by Western blotting analysis, which showed that the yield of the Follistatin cDNA was a 63kD protein expression vector. Follistatin protein was expressed in the form of glutathione-S-transferase (GST) fusion protein in E. coli.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; Follistatin ; chemistry ; genetics ; Molecular Sequence Data ; Phylogeny ; Recombinant Fusion Proteins ; biosynthesis ; Swine

AIMTo study the endogenous mechanism for the inhibition of aquaporin-1 expression in rat renal proximal tubule epithelial cells in response to acetazolamide.

METHODSPrimary cultured rat renal proximal tubule epithelia cells were divided into two groups: one was subjected to 1 x 10(-5) mol.L-1 acetazolamide, the other served as normal control. When grown to sub-confluency, the cells were disintegrated to perform isoelectrofocusing electrophoresis in order to find the differential proteins induced by the acetazolamide treatment. The differential proteins were defined by peptide mass fingerprinting technology.

RESULTSTwo differential proteins were found in the cell disintegrant. The pI 3.8 protein was reduced after treatment, which showed 21.4% similarity with the brush border membrane myosin from rat brain and testis, and 27% with glycogen phosphorylase; The pI 5.5 protein was increased on the contrary, with 20% similarity to phosphatidylinositol transfer protein alpha isoform.

CONCLUSIONAcetazolamide inhibited AQP1 expression probably by affecting the expression of pI 3.8 and pI 5.5 proteins.

Acetazolamide ; pharmacology ; Animals ; Aquaporin 1 ; Aquaporins ; antagonists & inhibitors ; metabolism ; Diuretics ; pharmacology ; Epithelial Cells ; metabolism ; Isoelectric Focusing ; Kidney Tubules, Proximal ; cytology ; metabolism ; Male ; Peptide Mapping ; Rats ; Rats, Sprague-Dawley

Aflatoxins, found in contaminated food, are potent hepatocarcinogen. The aflatoxin-detoxiczyme (ADTZ) isolated from the edible fungus Armillariella sp., detoxifies aflatoxin B1 (AFB1). This paper reports on the characterization of immobilized ADTZ using a hydrophobic adsorption method. The ADTZ was isolated from cryo-homogenated fungus, previously cultivated at 24 - 28 degrees C for 20 - 30 days, using n-alkyl amino-agar beads. Various adsorption conditions of the enzyme to n-alkyl or n-octyl amino-agar beads were carried out. The effects of enzyme immobilization on different alkyl amino-agar beads, at different pH values (5.5 - 7.5), at different temperature (20 - 40 degrees C) and at different salt concentrations were investigated. The enzyme activity was measured at OD360 by reacting 133.3 ng/mL of AFB1 at 30 degrees C for 30 min with the immobilized ADTZ. The Km value of the immobilized enzyme, determined using Schematic Linewearver-Burk plot, is 3.308 x 10(-3) mol/L, lower than that of free enzyme, which is 2.16 x 10(-6) mol/L. This indicated the affinity of the detoxiczyme to AFB1 decreased after immobilization. The immobilized enzyme activity in oil-phase (n-hexane) was also studied with different concentration of water. After the treatment of the immobilized ADTZ, the toxin no longer causes liver toxicity in the rat toxicity test, no longer causes mutagenicity in Ames test and is no longer toxic in the chicken embryo test. Results also indicated that the pH stability, the thermostability and the freezing stability of ADTZ were improved after the immobilization.
Absorption ; Aflatoxin B1 ; metabolism ; toxicity ; Animals ; Chickens ; Enzyme Stability ; Enzymes, Immobilized ; chemistry ; metabolism ; Hydrogen-Ion Concentration ; Hydrophobic and Hydrophilic Interactions ; Multienzyme Complexes ; chemistry ; metabolism ; Rats ; Temperature ; Toxicity Tests

Phytase gene of Aspergillus niger 963 was cloned into baculovirus transfer vector. DNA of the recombinant vector was co-transfected with Bm-BacPAK6 DNA into BmN cells, and recombinant virus was selected by plaque assays. The recombinant virus was identified by Dot blot and Southern-blot with the specific probe for phytase gene. Phytase gene was expressed in silkworm larvae and pupae. The expression product was 1.43 g/L haemolymph for silkworm larvae and 1.90 g/L haemolymph for pupae, respectively. The enzymic characteristicses of phytase expressed in baculovirus-expression system were studied in this paper.
6-Phytase ; genetics ; metabolism ; Animals ; Aspergillus niger ; enzymology ; genetics ; Blotting, Southern ; Bombyx ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Fungal Proteins ; genetics ; metabolism ; Genetic Vectors ; genetics ; Hydrogen-Ion Concentration