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

Objective:This work aims to investigate diallyl disulfide (DADS) inhibition of cell migration and invasion in human colon cancer SW480 cells through the Rac1-ADF/cofilin1 pathway. Methods:The potential of cell migration and invasion was examined by scratch healing assay and transwell membrane assay. The expression of Rac1-ADF/cofilin1 pathway was detected by RT-PCR and Western blot. Results:After the SW480 cells were treated with 40 and 50 mg·L-1 of DADS for 24 h, the number of transmembrane cells through the Matrigel obviously decreased by 57.12%and 64.59%, respectively (P<0.05). After cell treatment for 48 h, the cell migration rates were 23.23%and 12.87%, which were significantly lower compared with the control group (75.86%;P<0.05). After the cells were treated with 45 mg·L-1 of DADS for 24 and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and destrin mRNA respectively decreased compared with the control group (P<0.05). However, no significant difference was observed in the expression of cofilin1 mRNA (P>0.05). After the treatment with 45 mg·L-1 of DADS for 6, 12, 24, and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and Destrin proteins respectively decreased in a time-dependent manner compared with the control group (P<0.05). However, no significant differences were observed in the expression of the cofilin1 protein (P>0.05). Moreover, the expression of p-LIMK1 and p-cofilin1 notably decreased in a time-dependent manner (P<0.05). Conclusion:DADS inhibits cell migration and invasion, which is related to the down-regulation of Rac1, Rock1, PAK1, LIMK1, p-LIMK1, p-cofilin1, and destrin through the Rac1-ADF/cofilin1 pathway.

Adult ; Aortic Valve ; microbiology ; pathology ; Autopsy ; Brain ; microbiology ; pathology ; Endocarditis, Bacterial ; complications ; microbiology ; pathology ; Female ; Heart Ventricles ; microbiology ; pathology ; Humans ; Mitral Valve ; pathology ; Sepsis ; complications ; microbiology ; pathology ; Substance Abuse, Intravenous ; complications ; microbiology ; pathology ; Young Adult

Animals ; Collagen Type IV ; metabolism ; Fibronectins ; metabolism ; Glomerulonephritis, Membranous ; metabolism ; pathology ; Immune Sera ; immunology ; Kidney Glomerulus ; pathology ; ultrastructure ; Male ; Malondialdehyde ; blood ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; pharmacology ; Superoxide Dismutase ; blood ; Taurine ; pharmacology ; Transforming Growth Factor beta1 ; metabolism

BACKGROUNDPneumocystis jirovecii pneumonia (PCP) is one of the most common and fatal infections in non-AIDS immunocompromised patients, which is difficult to diagnose by traditional morphologic methods. This study evaluated polymerase chain reaction (PCR) assays of Pneumocystis jirovecii mitochondrial large subunits ribosomal RNA in sputum and bronchioalveolar lavage fluid (BALF) for diagnosing PCP.

METHODSSputum and BALF specimens from two groups were collected: one group (PCP group) included 20 patients definitely diagnosed of PCP by Gomori methenamine silver (GMS) stains of BALF; the other group (non-PCP group) included 40 patients. Each specimen was examined by GMS stains and PCR assays.

RESULTSGMS stains of BALF in PCP group were 100% positive (20/20), GMS stains of sputum in PCP group were 35% positive (7/20); GMS stains of BALF in non-PCP group were 100% negative (40/40), GMS stains of sputum in non-PCP group were 100% negative (40/40). PCR assays of BALF in PCP group were 100% positive (20/20), PCR assays of sputum in PCP group were 100% positive (20/20); PCR assays of BALF in non-PCP group were 100% negative (40/40), PCR assays of sputum in non-PCP group were 100% negative (40/40). Sensitivity and specificity of PCR assays of sputum and BALF were both 100%; positive and negative predictive values were also both 100%.

CONCLUSIONThe diagnostic value of PCR assays of Pneumocystis jirovecii mitochondrial large subunits ribosomal RNA on sputum and BALF for pneumocystis pneumonia are both high and equivalent.

Acquired Immunodeficiency Syndrome ; diagnosis ; genetics ; Bronchoalveolar Lavage Fluid ; chemistry ; Humans ; Pneumonia, Pneumocystis ; diagnosis ; genetics ; metabolism ; Polymerase Chain Reaction ; methods ; Sputum ; chemistry

OBJECTIVETo explore the effect of Chang'an No. I Recipe (CA) on 5-hydroxytryptamine signal system and mRNA expression levels of hippocampal brain derived neurotrophic factor (BDNF) in visceral hypersensitivity model rats with irritable bowel syndrome (IBS).

METHODSIBS visceral hypersensitivity rat models were established by combined chronic restraint stress and forced swimming. Successfully modeled rats were randomly divided into the model group, the Dicetelgroup (27 mg/kg) , the Fluoxetine group (3.6 mg/kg), the high dose CA group (22.6 mg/kg), the medium dose CA group (11.3 mg/kg), and the low dose CA group (5.7 mg/kg) according to body weight, 9 in each group. Besides, a normal control group with 10 rats was set up. Corresponding medication was administered to rats in each treatment group. Equal volume of physiological saline was administered to rats in the model group by gastrogavage. All medication was performed once per day for a total of 14 days. Pain threshold was determined by abdominal withdrawal reflex (AWR). Changes of colon 5-HT levels were determined by immunohistochemical assay. mRNA expression levels of hippocampal 5-hydroxytryptamine 1A receptor (5-HT1a) and BDNF were detected by immunofluorescent RT-PCR.

RESULTSCompared with the normal control group before treatment, pain threshold was obviously lowered in proctectasia rats of each group (P < 0.01). Compared with the normal control group after treatment, pain threshold was obviously lowered in rats of the model group; colon 5-HT levels, mRNA expression levels of hippocampal 5-HT1a and BDNF were obviously elevated (P < 0.01). Compared with the model group, pain threshold was obviously elevated in the Fluoxetine group and all CA groups; colon 5-HT levels were obviously reduced in the Dicetel group, high and medium dose CA groups (P < 0.05, P < 0.01); mRNA expression levels of hippocampal 5-HT1a and BDNF were obviously reduced in each CA group (P < 0.01); mRNA expression levels of hippocampal BDNF were obviously reduced in the Fluoxetine group (P < 0.01).

CONCLUSIONSThe target points of CA were involved in brain and gut. CA could reduce pain threshold of proctectasia rats, down-regulate colon mucosal 5-HT levels, and lower mRNA expression levels of BDNF and 5-HT1a in rat hippocampus.

Animals ; Brain-Derived Neurotrophic Factor ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hippocampus ; Hypersensitivity ; Intestinal Mucosa ; Irritable Bowel Syndrome ; drug therapy ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serotonin ; metabolism

Objective To investigate the expression of JAM-1 after microwave irradiation and its correlation with blood-brain barrier integrity. Methods A total of 160 male Wistar rats were divided into a sham radiation group and a radiation group. The radiation group was subdivided into three sub-groups treated with micrewaves at average power densities of 10, 30 and 100 mW/cm2. Rats in each group were sacrificed and their brain tissue sampled at 6 hours and 1, 3, 7 and 14 days after the irradiation. Evans blue ( EB ) dye, laser confocal microscopy,Western blotting, RT-PCR and image analysis were used to test the permeability of the blood-brain barrier and the expression of JAM-1 in protein and at the gene level in the rats' hippocampus and cortex. Results There was an increase of EB in the hippocampus 3 to 14 days after 10 and 100 mW/cm2 microwave irradiation. The EB level increased progressively in the 10 and 30 mW/cm2 groups within 7 d after irradiation but recovered by the 14th day. It also increased progressively in the 100 mW/cm2 group within 14 d after irradiation. In the hippocampus, EB was observed only in the lumens of the blood vessels in the sham group, but EB had diffused out of the blood vessels in the irradiated groups by the 3rd day after irradiation. After 10 or 30 mW/cm2 microwave irradiation, JAM-1 protein in the hippocampus and cortex decreased significantly within 7 d after irradiation but recovered by the 14th day. It decreased progressively in the 100 mW/cm2 group within 14 d after irradiation. The expression of JAM-1 mRNA in the hippocampus decreased significantly at 6 h after irradiation at all power levels, but it recovered within 7 days in the 10 and 30 mW/cm2 groups. Conclusions Microwave radiation can decrease the expression of JAM-1 in the hippocampus and cortex. The degree of decrease is positively correlated with the microwave radiation power. The change might involve increasing the permeability of the blood-brain barrier.

The 1H-NMR fingerprints of three different species tibetan medicine sea buckthorn were established by 1H-HMR metabolomics to find out different motablism which could provide a new method for the quality evaluation of sea buckthorn. The obtained free induction decay (FID) signal will be imported into MestReNova software and into divide segments. The data will be normalized and processed by principal component analysis and.partial least squares discriminant analysis to perform pattern recognition. The results showed that 25 metabolites belonging to different chemical types were detected from sea buckthorn,including flavonoids, triterpenoids, amino acids, carbohydrates, fatty acids, etc. PCA and PLS-DA analysis showed three different varietiest of sea buckthorn that can be clearly separated by the content of L-quebrachitol, malic acid and some unidentified sugars, which can be used as the differences metabolites of three species of sea buckthorn. 1H-NMR-based metabonomies method had a holistic characteristic with sample preparation and handling. The results of this study can offer an important reference for the species identification and quality control of sea buckthorn.
Hippophae ; metabolism ; Magnetic Resonance Spectroscopy ; methods ; Medicine, Tibetan Traditional ; Metabolomics

OBJECTIVETo evaluate the value of cytomorphologic and immunocytochemical approaches in the diagnosis of hematologic neoplasms in serous effusion.

METHODSThe cytospin and Thinprep smears of effusion specimens were prepared from 23 cases of lymphoid malignancies with histological confirmation and 30 cases of benign effusions used as control. Morphological assessment of the cellular components was conducted, including the ratio of mesothelium to lymphocyte, karyomorphism of lymphoid cell and the presence of apoptosis and mitosis. Immunocytochemical study was performed in all the cases, with flow cytometry in one case.

RESULTSAmong the 23 tumor cases, 14 represented disease relapse, and in the remaining nine cases, the serous effusion was the primary manifestation. The proportion of mesothelium was low in the tumor group, being less than 10% in 20 cases (87.0%, 20/23). It was more than 10% in most of benign cases (20/30, 66.7%). Lymphoid cells were prominent (> 80% cells) in 69.6% of the tumor cases, and the cellular component in some control cases (63.3%, 19/30) showed fewer lymphocytes. Nipple-like projection of lymphocytic nuclei could be detected in almost all the tumor cases (91.3%, 21/23), but was occasionally found in the control group (26.7%, 8/30). Apoptosis and mitosis were obvious in lymphomatous effusion, but observed in only 6.7% of the control cases. Significant difference of the previously mentioned cytomorphologic features existed between the tumor and control groups (P < 0.01). The results of immunocytochemical staining in cell block were identical to the corresponding immunohistochemistry, and one case of mantle cell lymphoma was confirmed by flow cytometry. The cytologic findings seen in all the 23 studied cases were in agreement with the corresponding histologic diagnosis.

CONCLUSIONSSome cytomorphologic features, including decreased number of mesothelium, increased number of lymphoid cells, nuclear nipple-like projection, and the presence of apoptosis and mitosis, are very useful for diagnosing lymphoid malignancy in serous effusion. Immunocytochemistry is an important approach to the cytodiagnosis and classification of lymphoma.

Adult ; Aged ; Aged, 80 and over ; Apoptosis ; Ascitic Fluid ; pathology ; Cyclin D1 ; metabolism ; Cytodiagnosis ; methods ; Female ; Humans ; Immunohistochemistry ; Interferon Regulatory Factors ; metabolism ; Lymphocytes ; pathology ; Lymphoma ; complications ; metabolism ; pathology ; Lymphoma, Large B-Cell, Diffuse ; complications ; metabolism ; pathology ; Male ; Middle Aged ; Mitosis ; Pleural Effusion, Malignant ; etiology ; metabolism ; pathology ; Young Adult

BACKGROUNDHydrogen sulfide (H 2 S) plays a protective role in chronic hemodialysis (CHD) patients. In this study, we further investigate the relationship between H 2 S and conventional protein kinase CβII (cPKCβII) in CHD patients with uremic accelerated atherosclerosis (UAAS).

METHODSA total of 30 healthy people, 30 CHD patients without AS and 30 CHD patients with AS (CHD + AS) were studied. Plasma H 2 S was measured with a sulfide sensitive electrode, and cPKCβII membrane translocation was detected by Western blotting.

RESULTSPlasma H 2 S in CHD + AS group was significantly lower than that in CHD patients. cPKCβII membrane translocation in CHD + AS group increased significantly compared with CHD group. Plasma H 2 S concentration was negatively correlated with cPKCβII membrane translocation in CHD + AS patients.

CONCLUSIONSThese findings suggest a possible linkage between H 2 S metabolism and cPKCβII activation, which may contribute to the development of UAAS in CHD patients.

Adult ; Aged ; Atherosclerosis ; blood ; enzymology ; metabolism ; Blotting, Western ; Humans ; Hydrogen Sulfide ; blood ; Middle Aged ; Protein Kinase C ; metabolism ; Renal Dialysis ; Young Adult