Objectiive:The present study was designed to explore whether overexpression of human wild ?-synuclein in rat brain caused selective dopaminergic neuron loss in substantia nigra and aimed to find out a new method to make a rat model of Parkinson's disease(PD).Methods:The human wild ?-synuclein gene was induced into the rat brain by Adeno-Associated Virus(AAV) vector.The overexpression of ?-synuclein was detected by realtime PCR.The behavior of rats were recorded every 4 weeks after the viral particle injection.TH immunohistochemistry were performed at 4,8,12 and 16 weeks post-injection as well as the dopamine(DA),3,4-dihydroxypheny-lacetic acid(DOPAC) of striatum were determined by high performance liquid chromatography coupled with electrochemical detection.Results:Realtime PCR results revealed a significant overexpression of ?-synuclein in the injected hemisphere.By 8 weeks post injection,a significant loss of the dopaminergic neurons was observed.34% of the dopaminergic neurons were lost after 12 weeks,and about 60% cells loss after 16 weeks.The DA and DOPAC levels in the striatum decreased about 15% 12 weeks after injecting viral particle carried ?-synuclein gene and 30% decreased after 16 weeks.The AAV-?-synuclein-treated rats developed a type of motor impairment,i.e.,head position bias,compatible with this magnitude of nigrostriatal damage.Conclusion:All the results showed that overexpression of human wild ?-synuclein caused selective dopaminergic neuron loss and mimic a symptom of human PD in rats.This may be a new methed to make rat PD model which can offer new opportunities for the study of pathogenetic mechanismsand exploration of new therapeutic targets of particular relevance to human PD.

Objective To understand the distribution of the river beach wetlands and Oncomelania snails in the lower reach-es of the Yangtze River,and explore the countermeasures of snail control. Methods The river beach wetlands outside the Yang-tze River levee were investigated and classified according to the hierarchical and classification system of wetlands of China. The snail survey was carried out in the beach wetlands of Runzhou section of lower reaches of the Yangtze River from 2004 to 2013. The change trend of snail areas and the densities was analyzed in the wetlands. Results The river beach of Runzhou section of lower reaches of the Yangtze River belongs to the riverine wetland. There was Oncomelania snail breeding except the permanent wa-ter area. At present,there were natural wetlands of 1 303.0 hm2,human-made wetlands of 1 479.0 hm2 and wetland function chang-es of 1 059.0 hm2 in the river beach of Runzhou section. There was the snail area of 181.4 hm2 in the natural wetland in 2013. The area of snail control by the molluscicide and environmental modification was 4 624.55 hm2 from 2004 to 2013. The decline rates of snail areas and densities were 66.53%and 77.66%respectively. The existing Oncomelania snails were distributed in the natural wetlands. Conclusion The human-made wetland is helpful to snail control. The snail control in the river beach wetlands should attach a great importance to the protection of wetland ecology.

Objective To explore the relationship between serum homocysteine (Hcy) level before coronary angiography(CAG) and contrast induced nephropathy (CIN) after CAG.Methods We included 2264 cases of suspected coronary heart disease from May 2013 to May 2016 and all patients received CAG examination.According to whether CIN has developed or not after CAG, the patients were divided into the non-CIN group (n=2162) and the CIN group (n=102).We analyzed and compared the clinical baseline data, serum Hcy and creatinine (Cr) levels and the estimated glomerular filtration rate between the 2 groups eGFR.Results Patients in the non-CIN group were younger and with less comorbidities of diabetes and chronic kidney disease (all P<0.05).The volume of contrast media consumed in the non-CIN group was less than the CIN group [(122±21)ml vs.(147±24)ml, P=0.012).Hcy level in the non-CIN group (12.81±6.71) μmol/L was lower than that in the CIN group (21.74±11.9)μmol/L before CAG (P<0.05).No significant differences in serum Cr level and eGFR before CAG (P>0.05).At 72 hours after CAG, Cr level of the non-CIN group (69.34±19.54 μmol/L) was lower than that of the CIN group (87.34±21.38) μmol/L (P<0.05).eGFR was higher in the non-CIN group (79.34±19.54)ml/min than that in the CIN group (67.34±21.38)ml/min (P<0.05).Linear regression analysis showed that Hcy level before CAG were positively correlated with Cr level after CAG (r=0.547,P<0.01) and negatively correlated with eGFR after CAG (r=-0.271,P<0.01).Conclusions Hcy level before CAG can be used as one of an effective parameter to predict CIN.

OBJECTIVE: To detect the levels of index related to inflammation such as soluble CD40 ligand (sCD40L), neutrophil collagenase-8 (MMP-8), and pregnancy associated plasma protein-A (PAPP-A) , lipid peroxidation and autoimmune indexes such as oxidized low density lipoprotein (ox-LDL) and its antibody (ox-LDL Ab) in patients with coronary heart disease, and to investigate its relationship with acute coronary syndrome (ACS). METHODS: Contents of sCD40L, MMP-8, PAPP-A, ox-LDL and ox-LDL Ab in the peripheral blood were measured by enzyme-linked immunosorbent assay from 109 patients with coronary heart disease including 36 acute myocardial infarction (AMI), 38 unstable angina pectoris (UAP), and 35 stable angina pectoris (SAP) and 36 controls without coronary heart disease. RESULTS: The levels of each index in the peripheral blood of ACS patients (including AMI and UAP) were higher than those of SAP patients and controls (P < 0.05), and the difference of each index between UAP group and AMI group in ACS patients had no statistical significance (P > 0.05). The levels of each index of SAP patients, except PAPP-A, were all higher than those of controls (P <0.05). All the indexes were helpful in diagnosis of ACS. The area under the ROC curve of each index is between 0.7 and 0.9. CONCLUSION The increase of sCD40L, MMP-8, PAPP-A, ox-LDL and ox-LDL Ab levels in peripheral blood may be related to the pathogenesis of ACS, and can be used as potential markers of unstable atherosclerosis plaque.
Aged ; Angina, Unstable ; blood ; immunology ; Autoantibodies ; blood ; Biomarkers ; blood ; CD40 Ligand ; blood ; Coronary Artery Disease ; blood ; immunology ; Female ; Humans ; Lipoproteins, LDL ; blood ; immunology ; Male ; Matrix Metalloproteinase 8 ; blood ; Middle Aged ; Myocardial Infarction ; blood ; immunology ; Pregnancy-Associated Plasma Protein-A ; metabolism

OBJECTIVEThis study sought to analyze the clinical manifestations and intervention of fulminant septic shock in community-acquired Pseudomonas aeruginosa septicemia.

METHODSWe retrospectively reviewed the medical records for diagnosis, antibiotic therapy, clinical course of septic shock, respiratory support, laboratory data etc.

RESULTSEight of nine cases with P. aeruginosa septic shock died. Fever (nine cases) and cough (three cases) or diarrhea (3 cases) were the 2 most common initial symptoms, three cases developed skin gangrenosum later. Pseudomonas aeruginosa infection was not considered in any of the cases before death or blood culture showed positive result. Only 3 cases were initially treated with susceptible antibiotic regimen but no anti pseudomonas combination therapy was applied, susceptible antibiotic monotherapy was applied in 7 cases after transfer to the ICU. The mean latency of shock occurrence was 5.1 hours (range 0 to 21 hours) after admission, the mean duration from the occurrence of shock to death was 13.8 hours (range, 1 - 32 hours). All the patients were transfer red to ICU for shock, the appropriate resuscitation of shock patients was delayed by 49.3 minutes (range 25 - 80 minutes) by transfer. Only two cases were diagnosed and treated for shock on admission; after transferred to ICU, only 5 patients were diagnosed as having shock, and only 3 received anti-shock treatment. Eight of the patients died of persistent shock. In 6 patients who died, mechanical ventilation was not applied until cardiac arrest occurred. All the patients had hypoalbuminaemia, elevated serum C-reactive protein concentration, leukopenia and 6 cases had DIC.

CONCLUSIONThe initial presentation of the cases with community-acquired Pseudomonas aeruginosa septicemia was nonspecific with fever and cough or diarrhea. Clinicians often underestimated the severity of the infection, few patients received effective antimicrobial therapy. The authors suggest that an anti-pseudomonas antibiotic should be included in the initial empiric antibiotic regimen to cover P. aeruginosa high-risk patients; the front-line clinician should be educated for early recognition and aggressive resuscitation of P infection. aeruginosa septicemia.

Adolescent ; Child, Preschool ; Community-Acquired Infections ; Female ; Humans ; Infant ; Male ; Pseudomonas Infections ; Pseudomonas aeruginosa ; Retrospective Studies ; Shock, Septic ; microbiology

OBJECTIVETo investigate the changes in cisplatin sensitivity of resistant ovarian cancer A2780 cells after inhibition of miR-23a expression and explore the molecular mechanisms.

METHODSThe drug-resistant ovarian cancer A2780 cells were exposed to cisplatin alone or in combination with antagomir-23a. The cell inhibition rates after the treatments were detected using MTT assay, cell cycle changes assessed with flow cytometry; and apoptotic cells observed using Hoechst33258 staining. The changes in glycoprotein P-gp expression in the cells were detected using Western blotting.

RESULTSInhibition of miR-23 a combined with cisplatin treatment significantly increased the cell inhibition rate (P<0.01) and lowered the IC(50) so of cisplatin by 83.76% from 110.18 μmol/L in the control group to 17.89 μmol/L (P<0.01). The combined treatments also caused cell cycle arrestin G0/G1 phase, increased the cell apoptosis rate (P<0.01) and the number of cells stained with Hoechst33258; the cellular expression of P-gp protein was significantly reduced as the cisplatin doses increased (P<0.01).

CONCLUSIONInhibition of miR-23a expression increases the sensitivity of A2780 cells to cisplatin possibly by inhibiting the negative regulation by miR-23a target genes that causes inhibition of P-gp protein expression.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; Cell Cycle ; Cell Line, Tumor ; drug effects ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; Female ; Humans ; MicroRNAs ; metabolism ; Ovarian Neoplasms ; pathology

Objective To explore the current status of the prenatal care quality and the influencing factors in Hangzhou City. Methods We used sociodemographic characteristics questionnaire and the Chinese version of QPCQ to investigate 383 pregnancy women who were at 36 weeks' gestation or greater by convenient sampling method from September to December in 2016. Results A total of 350 questionnaires were valid. The average age of 350 samples was (29.71±3.67) years old. There were 231 primiparas which accounted for 66.00％. Among them, 196 (56.00％) were satisfied with the prenatal care they received, and 153 (43.70％) think general, and only 1 (0.30％) felt unsatisfied. The score of the Chinese version of QPCQ was 3.61±0.50. The item "My prenatal care professional (s) always answered my questions honestly" had the highest scores of 4.10±0.62, while "My prenatal care professional (s) did things hastily" had the lowest scores of 2.44±0.96 . By multiple linear regression analysis, those people who had more visiting time (β' = 0.21, P<0.01), less waiting time (β' = -0.18, P<0.01) and the cognition of the essentiality of pregnancy school (β' = 0.12, P=0.02) were more likely to give higher regards for the quality of prenatal care. Conclusion Sufficient time scored relative lower in the whole questionnaire. So, hospitals could take efforts to shorten the waiting time by promoting the hospital process optimization, making good use of this period time and other strategies, so as to improve the quality of prenatal care.

OBJECTIVETo recognize the clinical features of the enterovirus 71 (EV71) infection with pulmonary edema or pulmonary hemorrhage as a fulminant and often fatal illness.

METHODSWe retrospectively reviewed the medical records of the three cases with EV71 infection for clinical manifestation, laboratory data, medications, outcome etc.

RESULTSAll the cases were infants and they all died. These infants had no skin or mucosal lesions, however, they had sudden onset of cyanosis and tachypnea 1 to 2 days after the onset of the febrile disease with vomiting. All these 3 cases were misdiagnosed and were treated for shock on admission. Pulmonary hemorrhage was not considered in any of the cases on admission. All the cases received tracheal intubation when foamy secretions were discharged from mouth and nose of the patients and notable cyanosis was noted. After intubation, all had pink foamy fluid flew out from the endotracheal tube. The patients had hyperglycemia and limb weakness, two had tachycardia, and hypertension was found in one case. Chest X-ray showed bilateral or unilateral widespread air space opacity, but the cardiac size and shape were normal. All the patients had leucocytosis. EV71 infection was confirmed by detection of specific sequences of the virus in throat swab and tracheal secretions samples and in one case in cerebrospinal fluid sample.

CONCLUSIONPulmonary edema or pulmonary hemorrhage occurred in the 3 cases with EV71-infected infants. The initial presentation was often nonspecific with fever and vomiting, and sudden appearances of cyanosis, tachypnea, tachycardia, hypertension or hypotension, limb weakness may suggest pulmonary edema or hemorrhage. Excessive fluid resuscitation may deteriorate the illness, on the contrary, fluid restriction and inotropic agents, and early intubation with positive pressure mechanical ventilation may be the proper treatment.

Enterovirus A, Human ; Enterovirus Infections ; pathology ; Female ; Hemorrhage ; etiology ; virology ; Humans ; Infant ; Male ; Pulmonary Edema ; etiology ; virology ; Retrospective Studies

Objective To investigate the toxic effect of hemin on primary cultured neurons,astrocytes,and brain capillary endothelial cells (BCECs),and the damage effect of hemin with different concentrations on the above cells. Methods (1) Primary cultured neurons,astrocytes and BCECs from the cortex of rats were exposed to different doses of hemin for 2 h,and continue culture of these cells for 24 to 96 h after withdrawing hemin was performed; the cellular morphology was examined under phase-contrast microscope; cellular survival rate was measured with Alama blue staining; and the releasing rate of lactate dehydrogenasing (LDH) was detected with regular biochemical method. (2) Primary cultured cells were exposed to different doses of hemin for 2 h,and continue culture of the cells for 4 h was performed after washing out the hemin; and then,concentrated formic acid was employed to dissociate the cells, and heme content in dissociated cells was measured with spectrophotometer. (3) Primary cultured cells was exposed to different doses ofhemin for 30,60 and 120 min,respectively,and continue culture of the cells for 4 h was performed after washing out hemin; and then,intracellular Fe3+was examined with Prussian blue staining. Results (1) Cultured neurons were injured by a low dose ofhemin (5 mmol/L) with a decreased survival rate by 40.2％ and an increased LDH releasing rate by 22.2％; and the pathological changes of cellular morphology were severe after 24 h of exposure to hemin.Following the increased doses ofhemin and time of post-exposure,the cellular death and LDH releasing were increased,and the morphological changes of cells were much severe. (2) The low and medium doses of hemin (5 mmol/L and 25 mmol/L) did not induce cellular death, LDH releasing and morphological changes in astrocytes; and a high dose ofhemin (50 mmol/L) could induce a death rate of astrocytes decreasing by 52.4％, a LDH releasing rate increasing by 31％ and obvious morphological changes of astrocytes; however, the injured astrocytes could regenerate fluent cellular monolayer 96 h after exposing to high dose of hemin treatment.(3) Hemin with either low or high dose did not induce any changes in cellular survival,LDH releasing and cellular morphology of BCECs.(4) The heme content in cultured neurons was significantly higher than that in astrocytes and BCECs after hemin treatment for 2 h.(5) The blue Fe3+ stained granules appeared in neurons as early as 30 min after neurons being exposed to hemin, and Fe3+ stained positive cells in neurons were significantly higher than those in astrocytes and BCECs at any dose ofhemin and any time point ofhemin treatment. Conclusion Hemin is highly toxic to neurons, but it can only injure astrocytes at a high dose and it can not induce direct damage in BCECs; free hemin could rapidly enter and accumulate in neurons,but less accumulate in astrocytes and not accumulate in BCECs.

OBJECTIVETo investigate weather there is a toll-like receptor 4 (TLR4)-mediated myeloid differentiation factor 88 (MyD88)-dependent pathway in hippocampal neurons of rats and the probable role of the pathway in neuroinflammation.

METHODSTo establish the proper model, primarily cultured hippocampal neurons were treated with lipopolysaccharides (LPS), or pretreated with TLR4 antibody then co-treated with LPS. The expression of mRNA of MyD88 and TNF-alpha receptor associated factor 6 (TRAF6) were tested by RT-qPCR. The content of MyD88 and TRAF6 were tested by Western blot. The nuclear translocation of nuclear factor-kappaB/P65 (NF-kappaB/p65) was tested by immunofluorescence. The content of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and nitric oxide (NO) were tested by ELISA.

RESULTSLPS could increase MyD88 and TRAF6 mRNA, upregulate protein level of MyD88 and TRAF6 and increase the level of TNF-alpha, IL-1beta and NO in cell culture supernatant. LPS also could promote NF-kappa B/p65 translation to the nucleus. The pretreatment with TLR4 antibody reduced the translocation to nucleus for NF-kappaB/P65 and the contents of TNF-alpha, IL-1beta and NO in the culture supernatant.

CONCLUSIONThere is a TLR4-mediated MyD88-dependent pathway in hippocampal neurons. The activation of this pathway can increase the level of TNF-alpha, IL-1beta and NO in cell culture supernatant. TLR4-mediated MyD88-dependent pathway in hippocampal neurons participate in neuroinflammation, that means neurons are not passive in inflammation.

Animals ; Cells, Cultured ; Hippocampus ; cytology ; metabolism ; Interleukin-1beta ; metabolism ; Myeloid Differentiation Factor 88 ; metabolism ; Neuritis ; metabolism ; Neurons ; metabolism ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; TNF Receptor-Associated Factor 6 ; metabolism ; Toll-Like Receptor 4 ; metabolism ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism