OBJECTIVETo characterize dark-adapted and light-adapted oscillatory potentials (OPs) in human electroretinogram (EGR) elicited by flashing light stimulation of the same intensity.

METHODSDark- and light-adapted ERGs of normal eyes were studied. The frequency spectra of the extracted dark-adapted OPs and light-adapted OPs were analyzed by a fast Fourier transform. The peak frequency, latency and total power of the OPs were determined.

RESULTSThe averaged peak frequency, latency, and power of the dark-adapted OPs was 125.3∓9.93 Hz, 41.7∓3.56 ms, and 9.25∓5.55 (V·s)(2), as compared with 79.5∓6.79 Hz, 50.8∓5.36 ms, and 3.56∓2.18 (V·s)(2) for light-adapted Ops, respectively, showing significant differences in the parameters between dark- and light-adapted Ops (P<0.001).

CONCLUSIONSCompared with dark-adapted OPs, light-adapted Ops is characterized by a lower peak frequency and a lower power with a prolonged latency.

Adaptation, Ocular ; physiology ; Adult ; Dark Adaptation ; physiology ; Electroretinography ; methods ; Female ; Humans ; Male ; Oscillometry ; Retina ; physiology ; Young Adult

To determine heat-shock protein (Hsp)90 expression is connected with cellular apoptotic response to heat stress and its mechanism, chicken (Gallus gallus) primary myocardial cells were treated with the Hsp90 promoter, aspirin, and its inhibitor, geldanamycin (GA), before heat stress. Cellular viability, heat-stressed apoptosis and reactive oxygen species level under different treatments were measured, and the expression of key proteins of the signaling pathway related to Hsp90 and their colocalization with Hsp90 were detected. The results showed that aspirin treatment increased the expression of protein kinase B (Akt), the signal transducer and activator of transcription (STAT)-3 and p-IKKα/β and the colocalization of Akt and STAT-3 with Hsp90 during heat stress, which was accompanied by improved viability and low apoptosis. GA significantly inhibited Akt expression and p-IKKα/β level, but not STAT-3 quantity, while the colocalization of Akt and STAT-3 with Hsp90 was weakened, followed by lower cell viability and higher apoptosis. Aspirin after GA treatment partially improved the stress response and apoptosis rate of tested cells caused by the recovery of Akt expression and colocalization, rather than the level of STAT-3 (including its co-localization with Hsp90) and p-IKKα/β. Therefore, Hsp90 expression has a positive effect on cellular capacity to resist heat-stressed injury and apoptosis. Moreover, inhibition of Hsp90 before stress partially attenuated its positive effects.
Apoptosis* ; Aspirin ; Cell Survival ; Chickens* ; Heat Stress Disorders ; Heat-Shock Proteins* ; Hot Temperature* ; HSP90 Heat-Shock Proteins ; In Vitro Techniques* ; Proto-Oncogene Proteins c-akt ; Reactive Oxygen Species ; Transducers

OBJECTIVETo prepare a porcine acellular dermal matrix (PADM), and to optimize the interpore distance between PADM and co-grafted split-thickness autologous skin.

METHODSPorcine skin was treated with trypsin/Triton X-100 to prepare an acellular dermal matrix. Micropores were produced on the PADM with a laser punch. The distance between micropores varied as 0.8 mm, 1.0 mm, 1.2 mm and 1.5 mm. Full-thickness defect wounds were created on the back of 144 SD rats. The rats were randomly divided into 6 groups as follows, with 24 rats in each group. Micropore groups I -IV: the wounds were grafted with PADM with micropores in four different intervals respectively, and covered with split-thickness autologous skin graft. Mesh group: the wounds were grafted with meshed PADM and split-thickness autograft.

CONTROL GROUPwith simple split-thickness autografting. The gross observation of wound healing and histological observation were performed at 2, 4, 6 weeks after surgery. The wound healing rate and contraction rate were calculated.

RESULTSTwo and four weeks after surgery, the wound healing rate in micropore groups I and II was lower than that in control group (P < 0.05), but no obvious difference was between micropore groups I , II and mesh group (P > 0.05) until 6 weeks after grafting( P <0.05). The wound contraction rate in micropore groups I and II ([(16.0 +/- 2.6)%, (15.1 +/- 2.4)%] was remarkably lower than that in control group 4 and 6 weeks after grafting (P < 0.05), and it was significantly lower than that in mesh group [(19.3 +/- 2.4)%] 6 weeks after surgery (P <0.05). Histological examination showed good epithelization, regularly arranged collagenous fibers, and integral structure of basement membrane.

CONCLUSIONLaser micropore PADM (0.8 mm or 1.0 mm in distance) grafting in combination with split-thickness autografting can improve the quality of wound healing. PADM with laser micropores in 1.0 mm distance is the best choice among them.

Animals ; Dermis ; transplantation ; Lasers ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; methods ; Skin, Artificial ; Swine ; Transplantation, Heterologous

The human UDP-glucuronosyltransferase 1A1 (UGT1A1), one of the most essential conjugative enzymes, is responsible for the metabolism and detoxification of bilirubin and other endogenous substances, as well as many different xenobiotic compounds. Deciphering UGT1A1 relevance to human diseases and characterizing the effects of small molecules on the activities of UGT1A1 requires reliable tools for probing the function of this key enzyme in complex biological matrices. Herein, an easy-to-use assay for highly-selective and sensitive monitoring of UGT1A1 activities in various biological matrices, using liquid chromatography with fluorescence detection (LC-FD), has been developed and validated. The newly developed LC-FD based assay has been confirmed in terms of sensitivity, specificity, precision, quanti-tative linear range and stability. One of its main advantages is lowering the limits of detection and quantification by about 100-fold in comparison to the previous assay that used the same probe substrate, enabling reliable quantification of lower amounts of active enzyme than any other method. The precision test demonstrated that both intra- and inter-day variations for this assay were less than 5.5％. Further-more, the newly developed assay has also been successfully used to screen and characterize the regu-latory effects of small molecules on the expression level of UGT1A1 in living cells. Overall, an easy-to-use LC-FD based assay has been developed for ultra-sensitive UGT1A1 activities measurements in various biological systems, providing an inexpensive and practical approach for exploring the role of UGT1A1 in human diseases, interactions with xenobiotics, and characterization modulatory effects of small mole-cules on this conjugative enzyme.

OBJECTIVE: To study the clinical features of the diseases associated with aminoacyl-tRNA synthetases (ARS) deficiency. METHODS: A retrospective analysis was performed of the clinical and gene mutation data of 10 children who were diagnosed with ARS gene mutations, based on next-generation sequencing from January 2016 to October 2019. RESULTS: The age of onset ranged from 0 to 9 years among the 10 children. Convulsion was the most common initial symptom (7 children). Clinical manifestations included ataxia and normal or mildly retarded intellectual development (with or without epilepsy; n=4) and onset of epilepsy in childhood with developmental regression later (n=2). Some children experienced disease onset in the neonatal period and had severe epileptic encephalopathy, with myoclonus, generalized tonic-clonic seizure, and convulsive seizure (n=4); 3 had severe delayed development, 2 had feeding difficulty, and 1 had hearing impairment. Mutations were found in five genes: 3 had novel mutations in the AARS2 gene (c.331G>C, c.2682+5G>A, c.2164C>T, and c.761G>A), 2 had known mutations in the DARS2 gene (c.228-16C>A and c.536G>A), 1 had novel mutations in the CARS2 gene (c.1036C>T and c.323T>G), 1 had novel mutations in the RARS2 gene (c.1210A>G and c.622C>T), and 3 had novel mutations in the AARS gene (c.1901T>A, c.229C>T, c.244C>T, c.961G>C, c.2248C>T, and Chr16:70298860-70316687del). CONCLUSIONS A high heterogeneity is observed in the clinical phenotypes of the diseases associated with the ARS deficiency. A total of 14 novel mutations in 5 genes are reported in this study, which enriches the clinical phenotypes and genotypes of the diseases associated with ARS deficiency.
Amino Acyl-tRNA Synthetases ; genetics ; Child ; Epilepsy ; Humans ; Mutation ; Phenotype ; Retrospective Studies

OBJECTIVETo investigate changes in proliferative activity of myoblasts in skeletal muscle and potential role of phosphorylated Akt on it, so that a better understanding in mechanisms of skeletal muscle atrophy after burn injury will be got.

METHODSOne hundred and twenty Wistar rats were randomly divided into 2 groups: control and severe thermal injury group. Rats in severe thermal injury group were subjected to a 40% total body surface area full-thickness scald injury, and Tibialis Anterior (TA) muscles were collected on 0, 1, 4, 7, 10, 14 days post-injury. After muscle mass determined, immunohistochemical double staining was used for detection of Proliferative Cell Nuclear Antigen (PCNA) of myoblasts. Protein expression of total Akt and phosphorylated Akt was determined by Western Blot.

RESULTSBurn injury induced significant reduction of TA muscle mass and maximal reduction of it appeared by 4 days after injury (P < 0.01). Proliferative activity of myoblasts decreased significantly from the first day post-injury (P < 0.01) and increased slowly to basal level of controls after 7 days post-injury. The phosphorylated Akt was undetectable in both of controls and injured samples before 4 days but increased significantly after 7 days post-injury (P < 0.01), though total Akt expression had no significant alteration at any time points (P > 0.05).

CONCLUSIONSDecrease in proliferative activity of myoblasts may be one of the contributors of significant atrophy of skeletal muscle after burn injury. Effect of phosphorylated Akt on proliferation attenuated in early stage and increased significantly in later stage after burn injury may partly explain the changes in proliferative activity of myoblasts.

Animals ; Burns ; metabolism ; pathology ; Cell Proliferation ; Disease Models, Animal ; Male ; Muscle, Skeletal ; metabolism ; pathology ; Myoblasts ; metabolism ; pathology ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo investigate changes in apoptosis-related ligands in serum in rats with severe scald and the effect of intensive insulin therapy on the changes.

METHODSOne hundred and fifty Wistar rats were randomly divided into 3 groups: sham burn (SB), scald (S) and treatment (T) groups. Rats in S and T groups were inflicted with 40% TBSA full-thickness burn, followed by intraperitoneal injection with 40 mL/kg of isotonic saline for resuscitation. Rats in T group were subcutaneously injected insulin in a dose of 0.25 U/100 g 24 hours after burn injury, and every 12 hours for 5 days (0.25, 0.50, 0.75, 1.00, 1.25 U/100 g each day, respectively) to control the level of blood glucose between 3 and 6 mmol/L. Rats in SB group were sham scalded at 37 degrees C without resuscitation. Blood was drawn from abdominal aorta on 1, 4, 7, 10, 14 post burn day (PBD) for determination of serum levels of TNF-alpha, soluble Fas ligand (sFasL) and soluble Fas receptor (sFas) by enzyme-linked immunosorbent assay (ELISA), and insulin by radioimmunity assay (RIA).

RESULTSThe serum level of TNF-alpha in S group peaked on 1 PBD (30.9 +/- 8.7) ng/L, which showed statistically significant difference when compared with that of SB and T groups (12.7 +/- 2.8) ng/L, (16.8 +/- 4.7) ng/L, respectively, P < 0.01), then lowered gradually to become similar to that of SB group on 7 PBD. The level of TNF-alpha in T group increased gradually, but was obviously lower than that of S group on 1, 4, 7 PBD (P < 0.01). The level of sFasL in S (on 7-14 PBD) and T (4-10 PBD) groups was significantly higher than that in SB group (P < 0.05), then lowered to normal level. The levels of sFas on 4-10 PBD in T group were obviously higher than that in S and SB group (P < 0.05). Ratio of sFasL to sFas in serum of S group was higher than that in SB group on 7, 10 PBD, which was higher than that in T group on 7 PBD (P < 0.05). There was significant decrease in serum level of insulin in S group compared with that of SB group on 4-10 PBD (P < 0.05). The level of insulin in T group increased on 1 PBD, peaked on 4 PBD (327 +/- 15 microU/mL), which was significantly higher than that in SB and S groups (42 +/- 15, 28 +/- 10 microU/mL, respectively, P < 0.01), then decreased gradually to normal level.

CONCLUSIONSInsulin may inhibit apoptosis after burn by down-regulating secretion of apoptotic ligands.

Animals ; Apoptosis ; Blood Glucose ; analysis ; Burns ; blood ; drug therapy ; Fas Ligand Protein ; blood ; Insulin ; therapeutic use ; Male ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood ; fas Receptor ; blood

OBJECTIVETo explore the effects of percutaneous transhepatic radiofrequency ablation (PRFA) combined with tumor edge of percutaneous absolute ethanol injection (PEI) on liver cancer adjacent to major blood vessels.

METHODSSeventy five patients with liver cancer adjacent to major blood vessels were randomly divided into two groups: PRFA+PEI therapy group (38 cases) and PRFA control group (37 cases). Tumor necrosis rate, AFP levels, local recurrence rate, median for survival time and cum survival were used as the evaluation index to evaluate the efficacies of the two methods.

RESULTSTumor necrosis rates of the therapy group and the control group were 84.2% and 54.1% (P < 0.01), respectively; AFP levels of therapy group and control group at 1, 3, 6 and 12 months after treatment were (105.0 ± 35.5) μg/L, (28.4 ± 4.3) μg/L, (58.6 ± 6.7) μg/L, (89.5 ± 12.5) μg/L and (137.2 ± 34.6) μg/L, (84.2 ± 18.4) μg/L, (106.6 ± 20.3) μg/L, (173.7 ± 32.0) μg/L, respectively. The rates of therapy group was significantly lower than of control group. Local recurrence rates of the therapy group and control group were 2.6%, 7.9%, 13.2% and 31.6% vs 10.8%, 21.6% , 40.5% and 62.1% (P < 0.05) at 3, 6, 12 and 24 months after treatment, respectively. Median for survival time of the therapy group and control group were 28.0 ± 2.8 months and 19.0 ± 3.6 months, respectively. Cum survival of the therapy group and control group were 84.2%, 78.9%, 60.5% and 31.6% vs 78.4%, 67.6%, 37.8% and 8.1% (P < 0.05) at 6, 12, 24 and 36 months after treatment, respectively.

CONCLUSIONPEI as a supplementary treatment of PRFA can effectively improve the treatment of liver cancer adjacent to major blood vessels and significantly reduce the local recurrence rate and improve long-term survival rates.

Adult ; Aged ; Bile Duct Neoplasms ; Carcinoma, Hepatocellular ; pathology ; therapy ; Catheter Ablation ; Combined Modality Therapy ; Ethanol ; administration & dosage ; Female ; Humans ; Liver Neoplasms ; pathology ; therapy ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate ; Treatment Outcome

OBJECTIVESevere newborn hypoxic-ischemic encephalopathy (HIE) has a very high rate of disability and no effective treatment is available. The present study aimed to preliminarily evaluate the effects of human neural stem cell transplantation in treatment of severe neonatal HIE.

METHODSThe patient was a 75-day old male infant with sequelae of severe HIE who had highly delayed development of intelligence and movement and myotonia. MRI showed multiple cerebromalacia and encephalatrophy. Cells obtained from the forebrain of an 11-week old fetus were cultured and amplified for 15 days. And then the human fetal neural stem cells were injected into cerebral ventricle of this infant.

RESULTSTwenty eight days after transplantation, remarkable improvement occurred not only in his myotonia but also in his intelligence and movement, which became similar to those of the normal infants of the same age. Positron emission tomography (PET) showed significantly increased radioactivity at temporal and occipital lobes which suggested that the cellular metabolism had increased greatly.

CONCLUSIONThe short-term effect of NSCs transplantation on the infant with severe HIE sequelae was significant. PET suggested that the implanted NSCs survived. Many more studies are needed to evaluate long-term effects of NSC transplantation in treatment of HIE.

Asphyxia Neonatorum ; complications ; Brain ; pathology ; physiopathology ; Female ; Humans ; Hypoxia-Ischemia, Brain ; etiology ; pathology ; physiopathology ; therapy ; Infant ; Infant, Newborn ; Injections, Intraventricular ; Multipotent Stem Cells ; transplantation ; Neurons ; Positron-Emission Tomography ; Prognosis ; Stem Cell Transplantation ; methods ; Time Factors ; Treatment Outcome

OBJECTIVETo analyze the protein expression profiles of multinodular hepatocellular carcinoma (HCC) with multicentric occurrence (MO) or with intrahepatic metastasis (IM).

METHODS5 IM and 6 MO patients were divided into groups of IM1, IM2, MO1 and MO2 according to the size of node of HCC. Two dimensional gel electrophoresis (2-DE) and mass spectrum were used to analyze the protein expression profiles. Western blot was used to confirm the results obtained by mass spectrum.

RESULTS2-DE of IM1, IM2, MO1 and MO2 indicated that 30 protein dots were differentially expressed in these tumors. By mass spectrum, 25 proteins were identified. Gene ontology classification indicated that these proteins are associated to cell movement, signal transduction, oxidoreduction, lipid metabolism, and amino acid metabolism.

CONCLUSIONThe protein expression profiles of IM is different from that of MO, 2-DE and mass spectrum can be used to identify the molecular markers of IM and MO of HCC.

Adult ; Blotting, Western ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasms, Multiple Primary ; metabolism ; pathology ; Prognosis ; Proteome ; metabolism ; Proteomics