In practice of forensic medicine, potential disease can be associated with fatal asphyxia in restraint position. Research has demonstrated that nitric oxide (NO) and nitric oxide synthase (NOS) are plentifully distributed in skeletal muscle, contributing to the regulation of contractile and relaxation. In the current study, respiratory functions, indices of diaphragmatic biomechanical functions ex vivo, as well as NO levels in serum, the expressions of diaphragmatic inducible NOS (iNOS) mRNA, and the effects of L-NNA on contractility of the diaphragm were observed in sepsis induced by cecal ligation and puncture (CLP) under the condition of restraint position. The results showed that in the CLP12-18h rats, respiratory dysfunctions; indices of diaphragmatic biomechanical functions (Pt, +dT/dt(max), -dT/dt(max), CT, Po, force over the full range of the force-frequency relationship and fatigue resistance) declined progressively; the NO level in serum, and iNOS mRNA expression in the diaphragm increased progressively; force increased significantly at all stimulation frequencies after L-NNA pre-incubation. Restraint position 1 h in CLP12 h rats resulted in severe respiratory dysfunctions after relative stable respiratory functions, almost all the indices of diaphragmatic biomechanical functions declined further, whereas little change took place in NO level in serum and diaphragmatic iNOS mRNA expression; and the effects of L-NNA were lack of statistical significance compared with those of CLP12 h, but differed from CLP18 h group. These results suggest that restraint position and sepsis act together in a synergistic manner to aggravate the great reduction of diaphragmatic contractility via, at least in part, the negative modulation of NO, which may contribute to the pathogenesis of positional asphyxia.
Animals ; Asphyxia ; Diaphragm/physiology* ; Muscle Contraction ; Muscle, Skeletal ; Nitric Oxide/metabolism* ; Nitric Oxide Synthase ; Nitric Oxide Synthase Type II ; Rats ; Respiration Disorders ; Restraint, Physical ; Sepsis

The patient,a 11-year-old boy,presented with a 4-year history of erythema and vesicles on the face and arms as well as a 4-month history of tumor and ulcer on the extremities,accompanied by progressive fatigue and intermittent fever.The patient had a body temperature of 37.7℃.No lymph node involvement was observed.Cutaneous examination revealed minimally indurated pink-red patches on the face and nose and dusky red firm nodules and tumors of varying sizes on the extremities.The nodules ranged from 2.0 cm to 18 cm in diameter,some had necrosis and black crusts on the surface.Ulcers were observed in some of the larger nodules;many of the ulcers extended into the muscle layer.White purulent discharge was seen on the surface of many of the nodules.The lesions were sharply demarcated,firm,tender, and surrounded by small satelite nodules.Histologically,there were large quantities of irregularly shaped, middle-sized tumor cells with clear cytoplasm,large twisted nuclei and prominent chromatin,infiltrating from the epidermis to subcutaneous tissue.The tumor cells infiltrating the follicles and eccrine sweat glands were either distributed perivascularly in a nest shape,or dispersed.There were broken nuclei and reactive histio- cytic infiltration in the dermis and subcutaneous tissue.Immunohistologically,the tumor cells were positive for cytoplasmic CD3 around the nuclei,for CD56,CD45RO and T cell intracellular antigen-l,and partly for CD30,CD8 and Ki67.Epstein-Barr virus-encoded nuclear RNA was positive with in situ hybridization. TCR?-2 gene rearrangement was positive in these tumor cells.A diagnosis of hydroa vacciniforme-like primary cutaneous NK/T-cell lymphoma was made.Therefore,this is a case report of hydroa vaccini- forme-like primary cutaneous NK/T-cell lymphoma with primary involvement in the skin;the condition was slowly progressive over 51 months.

Objective To investigate the clinical diagnostic value and pathogenesis of serum protein identification in Keshan disease (KD).Methods A total of 65 chronic KD patients were selected as the patient group in KD endemic areas,while 29 cases of dilated cardiomyopathy (the DCM group),62 healthy cases from KD endemic areas (control 1 group) and 28 healthy cases from non-endemic areas (control 2 group) were selected as controls.Liquid chip time of flight mass spectrometry (ClinProtTM MALDI-TOF-MS) was used to determine the expression of proteins/peptide peaks.ClinProTools 2.2 software was used to analyze the protein profiles to determine differentially expressed proteins/peptide peaks.The Genetic Algorithm (GA),QuickClassifer Algorithm (QC) and Supervised Neural Network Algorithm (SNN) methods were used to screen marker proteins.Matrix-assisted laser desorption/ionization time-of-flight Mass Spectrometry technique (MALDI-TOF/TOF) was also used as a secondary mass spectrometry to identify differentially expressed peptides.Results Between the KD and control 1 groups,34 differentially expressed proteins/peptides and 5 marker proteins were identified,while 52 differentially expressed proteins/peptides and 5 marker proteins were identified between the KD and control 2 groups,and there were 67 differentially expressed proteins/peptides and 5 marker proteins between the KD and DCM groups.During secondary mass spectrometry,two peptides for mass-to-charge ratio (m/z) 2 079 and 1 465 were obtained,peptide of matching β-globin showed low expression while peptide of matching fibrinogen showed high expression in the KD patients.Conclusions Serum marker proteins can be used as biomarkers for diagnosis and differentiation of KD.β-globin and fibrinogen play an important role in the development of KD myocardial injury.

Objective To explore the relationship between the endemic arsenism and the liver,renal damage.Methods Some permanent residents were selected as investigated subjects who lived at 3 villages in Datong in Shanxi Province,an arseniasis-endemic areas,These objects were divided into arsenic poisoning and control group on the basis of Diagnosis Standard for Endemic Arsenism(WS/T 211-2001).Then blood and urine samples were collected in the surveyed people.Serum glutamate pyruvic transaminase(ALT)were detected by Enzyme-linked immunosorbent assay as the indicator of the impaired hepatic function.The microdosis albumen (mAlb)and acetylglucosaminidase(NAG)in urine were detected by end-point method and alkaline picric acid as the renal damage indicators.Results A total of 661 people investigated,of which 144 cases were arsenic poisoning patients.The rates of abnormal liver function were significant hisher in arsenic poisoning group[10.42% (15/144)]than that in control[5.22%(27/517)],and both wag significant[X2=5.107,P＜0.05;OR=2.11,95%CI (1.09-4.08)].The geometric mean of mAlb/Ucr was 2.16 mg/g Cr in control,and 2.31 mg/g Cr in arsenic poisoning group,and both was not significant(t=-1.71,P＞0.05).The geometric mean of NAG waft higher in arsenic poisoning group(2.43 U/g Cr)than that in the control(2.22 U/g Cr),and both was significant(t=-3.55, P＜0.05).Conclusions The damage of the liver and renal function were related with endemic arsenism,and NAG is the early indicators suggesting impaired renal function due to endemic arsenism.

Objective To analyze the characteristic colligation force vector curves in hemiplegic patients. Methods Eleven hemiplegic patients were recruited, and 11 healthy elderly served as control. The characteristic changes of the patients with regard to colligation force vector demonstrated on a feedback apparatus of the "X.O.S"system during the dynamic training exercise program were analyzed and compared with that of the control group. Results The value of the colligation force vector passing most of the regions were significantly decreased ( P

Background Posterior capsular opacification(PCO) is common complication after extrecapsular extract of cataract.Matrix metalloproteinases-3 (MMP-3) can degrade all the extracellular matrix except polyose.The gene therapy of PCO upon MMP-3 is the researching hot topic.Fibronectin ( FN ) is a degrade gelatin,so its expression can reflect the effect of MMP-3 on LECs indirectly. Objective The aim of this study was to construct MMP-3 eukaryotic recombination plasmid and transfect to lens epithelium cells(LECs) for the observation of MMP3 expression,and to explore the feasibility of gene therapy for after cataract. Methods Six fresh lenses were obtained from pigs.LECs were cultured using explant method.The eukaryotic expression vector pEGFP-N1-MMP-3 was reconstructed with MMP-3 and pEGFP-N1 plasmids.The accuracy of MMP-3 gene fragment was confirmed by double enzyme digestion and DNA sequencing analysis.After transfecting pEGFP-N1-MMP-3 into LECs of pig,the expression of MMP-3 protein in the cells was indirectly observed by green fluorescent protein.The expression of FN in LECs was detected using Western blot. Results The result of double enzyme digestion was consistent with the base number of pEGFP-N1 plasmids and target fragment.By enlacing the result of DNA sequencing analysis with software,the resemblance of the DNA sequence of MMP-3 from recombination plasmid pEGFP-N1-MMP-3 and that of homo MMP-3 was 99.6％,indicating that the target fragment was inserted to pEGFP-N1 plasmids successfully.Green fluorescence for GFP was seen in the LECs in pEGFP-N1-MMP-3 transfected group,but absent response for GFP was in empty vector group.Western blot revealed that the relative expression level of FN in LECs was 0.666±0.008 in pEGFP-N1-MMP-3 trasfected group and 0.326 ±0.071 in empty vector group,with a significant difference between these two groups(P=0.000). Conclusions Eukaryotic recombination plasmid pEGFP-N1-MMP-3 is successfully constructed,and MMP-3 can be expressed in LECs after transfected.These results lay a foundation for the further research of MMP-3 gene therapy for PCO.

AIM:To research the effects of lithium chloride on transforming growth factor beta (TGF-β) and connective tissue growth factor (CTGF) in cultured human Tenon capsule fibroblasts (HTFs) and explore its mechanism.METHODS:HTFs were cultured and identified by vimentin staining with immunofluorescence and the morphological characteristics.The experimental group was processed 48h with LiCl in concentration of 80mmol/L, the control group without LiCl.The mRNA expression of TGF-β and CTGF in two groups were analyzed with real-time fluorescent quantitative polymerase chain reaction (real time-qPCR) and the protein expression was detected with Western blot.RESULTS:The cultured HTFs expressed TGF-β and CTGF.The mRNA expression of TGF-β and CTGF significantly decreased compared with the control group(t=20.042, 14.995, P<0.05).the protein expression of TGF-β and CTGF also decreased significantly compared with the control group(t=46.058、12.452, P<0.05)CONCLUSION:The cultured HTFs can express TGF-β and CTGF in mRNA and proteins' level.LiCl can reduce the expression of TGF-β and CTGF both in gene and proteins' level.LiCl has the potential to modulate wound healing for glaucoma filtration surgery.

To observe BAG-1, EGFR, and PARP-1 expressions in invasive breast cancer and its correlation with clini-cal pathological indicators, as well as to evaluate their clinical significance. Methods:The BAG-l, EGFR, and PARP-1 expressions in a tissue microarray of invasive breast cancer and peritumoral tissues were detected through immunohistochemical staining. The clinical and pathological significance of BAG-1, EGFR, and PARP-1 were evaluated. Results:The BAG-1, EGFR, and PARP-1 expression lev-els are higher in invasive breast cancer tissues than in peritumoral tissues (P<0.05). BAG-1 expression in invasive cancer tissues is not related to age, tumor site, lymph node metastases, and clinical TNM staging of patients, but is related to size, grade, ER, PR, and HER-2 expressions and molecular subtype (P<0.05). EGFR expression is related to size, clinical TNM staging, and molecular subtype (P<0.05). PARP-1 expression is related to grade, lymph node metastases, ER, and molecular subtype (P<0.05). BAG-1 expression is not significantly correlated with EGFR and PARP-1 in all cases, but BAG-1 and PARP-1 expressions are positively correlated in tri-ple-negative breast cancer tissues (P<0.05). Results of the univariate analysis revealed that the BAG-1 and PARP-1 expressions and the molecular subtypes are associated with the prognoses of breast cancer patients. Multivariate analysis revealed that BAG-1 and PARP-1 expressions are factors that are independent of the prognosis. Conclusion: BAG-1, EGFR, and PARP-1 overexpressions in human breast tissues suggest that BAG-1, EGFR, and PARP-1 are related to breast cancer development. BAG-1, EGFR, and PARP-1 are poten-tial biomarkers of breast cancer diagnosis and prognosis.

Objective To evaluate the significance of human interleukin-31(IL-31)in the pathogenesis of atopic dermatitis and its correlation with pruritus in patients with atopic dermatitis(AD).Methods Twenty-two children with mild to severe atopic dermatitis and 22 age-matched healthy controls were included in this study.Patients and controls were randomly and equally assigned into stimulation and non-stimulation groups.Venous blood samples were obtained from all participants,peripheral blood mononuclear cells were isolated from these samples and cultured with(stimulation groups)or without(non-stimulation groups)staphylococcal enterotoxin B(SEB)for 24 hours.Then,the mRNA expression of IL-31 on PBMCs was assessed via real-time reverse transcription-PCR.ELISA was used to detect the total serum IgE level in these objects.The severity of AD in patients was rated according to scoring atopic dermatitis(SCORAD).The relationship between the mRNA expression of IL-31 and the level of serum total IgE.severity of atopic dermatitis,and degree of pruritus.was evaluated.Results The expression of IL-31 mRNA on non-stimulated PBMCs from patients was 23.2 folds as high as that from the healthy controls(P＜0.01).The stimulation with SEB upregulated the mRNA expression of IL-31 on PBMCs.and the increase on PBMCs from patients was 20.44 times of that from the controls.The total serum IgE level was 260.05 IU/mL(5.9-1131.01 IU/mL)and 17.7 IU/mL(5-140.7 IU/mL)in the Patients and controls respectively(P＜0.01).There was no significant correlation between the mRNA expression of IL-31 and disease severity or total serum IgE level(r=0.07.0.22respectively.both P＞0.05)in patients witll AD.Condusions IL.3 1 is involved in t11e pathogenesis of AD,which is unlikely to be IgE-dependent.SEB can induce the rapid expression of IL-31 on PBMCs of healthy human,and is an important modulator for the production of IL-31.

Paeoniflorin is the main active ingredient of Chinese herbaceous peony. This study is to investigate the protective effect of paeoniflorin (Pae) on acute brain damage induced by lipopolysaccharide (LPS) in mice. The mice were randomly assigned to the normal control, model control (LPS), as well as groups of paeoniflorin and lipopolysaccharide (Pae + LPS). Then the mice were administered intraperitioneally with normal saline or Pae (10, 30 mg · kg(-1)) once daily for 6 d. One hour after intrapertioneally treatment on the seventh day, each group were injected LPS (5 mg · kg(-1)) to establish the endotoxin lipopolysaccharide inflammation model except the normal group. The mice were sacrificed after 6 h and the brain homogenates were prepared and measured. The malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), hydrogen peroxide (H2O2), succinatedehydrogenase (SDH), Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase were dectected by the colorimetric method. The levels of HO-1 and Nrf2 protein in subcellular fractions of brain tissue were detected by Western blot. The results demonstrated that the administration with paeoniflorin reduced the levels of the MDA production; significantly increase the activities of antioxidant enzyme (SOD and GSH-PX). In addition, paeoniflorin could enhance the total antioxidant capacity, decrease the level of H2O2, and increase the activities of SDH, Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase. Furthermore, paeoniflorin can increase the expression of HO-1 and activate the nuclear transfer of Nrf2. Taking together, these findings suggest that paeoniflorin alleviate the acute inflammation in mice brain damage induced by LPS, which is related with its antioxidant effect and improvement of energy metabolism.
Animals ; Energy Metabolism ; drug effects ; Glucosides ; pharmacology ; Heme Oxygenase-1 ; genetics ; Lipopolysaccharides ; pharmacology ; Male ; Membrane Proteins ; genetics ; Mice ; Mice, Inbred BALB C ; Monoterpenes ; pharmacology ; Oxidative Stress ; drug effects ; Sodium-Potassium-Exchanging ATPase ; metabolism