The present study is to investigate whether the extracellular signal-regulated kinase (ERK) and cAMP response element binding protein (CREB) signaling pathway contributes to the initiation of chronic constriction injury (CCI)-induced neuropathic pain in rats. Mechanical allodynia was assessed by measuring the hindpaw withdrawal threshold in response to a calibrated series of von Frey hairs. Thermal hyperalgesia was assessed by measuring the latency of paw withdrawal in response to a radiant heat source. The expressions of phosphor-ERK (pERK) and phosphor-CREB (pCREB) were examined using Western blot analysis and immunohistochemistry. An early robust increase in the expression of pERK on the spinal cords ipsilateral to injury was observed on day 1 after CCI, when the CCI-induced behavioral hypersensitivity had not developed yet. Moreover, the upregulation of pERK expression in ipsilateral spinal cord was associated with the increase in pCREB expression in bilateral spinal cord. Intrathecal administration of mitogen-activated protein kinase kinase (MEK) inhibitor U0126 before CCI can efficiently block and delay the CCI-induced mechanical allodynia and thermal hyperalgesia. These data suggest that activation of ERK and CREB in the spinal cord contributes to the initiation of peripheral nerve injury-induced pain hypersensitivity, and an early intervention strategy should be proposed.
Animals ; Butadienes ; pharmacology ; Cyclic AMP Response Element-Binding Protein ; metabolism ; Enzyme Inhibitors ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Hyperalgesia ; etiology ; physiopathology ; prevention & control ; Male ; Nitriles ; pharmacology ; Pain ; etiology ; physiopathology ; prevention & control ; Peripheral Nerve Injuries ; complications ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Sciatic Neuropathy ; metabolism ; physiopathology ; Spinal Cord ; metabolism

Abstract?AIM: To investigate mechanism of bradykinin ( BK) on inflammations of retinal pigment epithelium ( RPE) cells.?METHODS: ARPE -19 cells were cultured in vitro, stimulated by 100nM BK for 24h. Cell morphology changes were observed by microscope, and BK receptor localization was detected through cell immunofluorescence. Changes of Ca2+in BK and BR antagonist stimuli were detected by laser scanning confocal microscopy.The expressions of COX-1, COX-2, eNOS and iNOS protein in control group and BK group were detected by Western Blot.?RESULTS: After the stimulation of BK, there was no significant changes of ARPE-19 cells in morphology.Kinin B1 receptors ( B1R ) and B2 receptors ( B2R ) could be detected in ARPE-19 cells.Compared with control group, Ca2+concentrations significantly increased in BK group; in B1R antagonist group and B2R antagonist group Ca2+concentrations increased less than BK group; B1R and B2R antagonist group showed no obvious changes in Ca2+concentrations.Compared with control group, COX-2 and iNOS protein concentrations were significantly increased in BK group (P<0.001).?CONCLUSION:BK induces the increasing expression of COX-2 and iNOS in the cultured ARPE cells through binding with either B1R or B2R.

Objective:To explore the clinical effects of the "Disease-Characteristics" project in improving the nursing quality of patients with diabetic retinopathy (DR) .Methods:Based on the content framework of "Disease-Characteristics", authoritative data of UpToTate and various guidelines were transformed into evidence for clinical application through the steps of evidence-based nursing, and a standardized "Disease-Characteristics" project for DR patients was constructed. Convenience sampling was adopted to select nurses who worked in the Shanghai Eye Hospital and the Ophthalmology Department of Shanghai First People's Hospital from January 1, 2018 to June 30, 2019 as research nurses. Convenience sampling was used to select 223 DR patients admitted to the two hospitals from October 30, 2018 to January 31, 2019 (before implementation) as the control group, and 261 patients with DR admitted from February 1, 2019 to April 30, 2019 (after implementation) as the observation group. We observed and compared indicators such as the correct rate of nurses' assessment of each key link of DR patients before and after implementation, the awareness rate of patients' DR-related knowledge, and the incidence of postoperative complications of patients.Results:After the implementation, the scores of the nurses in the assessment of each key link of DR patients increased from (90.36±1.16) to (96.72±1.13) , and the difference was statistically significant ( t=52.990, P<0.05) . The disease-related knowledge score of DR patients increased from (74.330±12.154) to (78.386±10.094) , and the difference was statistically significant ( t=4.010, P<0.05) . Postoperative complications such as persistent high intraocular pressure, vitreous rebleeding, inflammatory edema and diplopia were improved, and the differences were statistically significant ( χ2=7.131, 5.618, 5.788, 6.128; P<0.05) . Conclusions:The "Disease-Characteristics" project plan based on the best evidence has a certain significance for improving the nursing quality of DR and the prognosis of patients.

OBJECTIVETo establish a loop-mediated isothermal amplification (LAMP) method for rapid diagnosis of Vibrio cholerae.

METHODSBased on the ompW nucleic sequence of Vibrio cholerae, a pair of primers was designed for LAMP. The reaction conditions were optimized, and the specificity, sensitivity, and practicability of LAMP were tested using 47 bacterial strains and simulated contaminated sites.

RESULTSThe results of viable bacterium count showed that LAMP was capable of detecting Vibrio cholerae at a level as low as 1.6x10(2) cfu/ml. The minimal detectable concentration was 1.6+10(3) cfu/ml for simulated contaminated samples such as feces and seawater, and 1.6+10(4) cfu/ml for contaminated milk. All the 21 strains of Vibrio cholerae yielded positive results in LAMP, and the 26 strains of other bacteria all showed negative results, with a detection specificity of 100%.

CONCLUSIONThe established LAMP method has high specificity and sensitivity for detecting Vibrio cholerae and is applicable in field monitoring and epidemiological study of Vibrio cholerae.

Bacterial Proteins ; genetics ; Cholera ; diagnosis ; microbiology ; Clinical Laboratory Techniques ; methods ; Humans ; Nucleic Acid Amplification Techniques ; methods ; Sensitivity and Specificity ; Vibrio cholerae ; genetics ; isolation & purification

OBJECTIVETo identify the mutation of low density lipoprotein receptor(LDLR) gene in a large Chinese family with familial hypercholesterolemia(F H) and make a discussion on the pathogenesis of FH at the molecular level.

METHODSInvestigations were made on a patient with the clinical phenotype of homozygous FH and his parents for mutations of promoter and all 18 exons of LDLR gene. Screening was carried out using Touch down PCR and a g arose gel electrophoresis, combined with DNA sequence analysis. The results were compared with the normal sequences in GenBank and FH database (www.ucl.uk/fh) t o find the mutation. Then the mutation was identified in other members of the family. In addition, the authors screened the apolipoprotein B(100) (apoB(100)) gene f or known mutations (R3500Q) that cause familial defective apoB(100) (FDB) by PCR-RFLP.

RESULTSA novel homozygous IN III 5' GT --> AT mutation in the splice donor of LDLR intron 3 was detected in the homozygote propositus with FH. The mutation was also identified in four heterozygous carriers in his family. No mutations R3500Q of apoB(100)were observed.

CONCLUSIONA homozygous G --> A splice mutation in LDLR gene was first reported. The change of the splice donor in LDLR intron 3 may cause skipping of exon 3, which is responsible for FH. Perhaps it is a particular pathogenesis for Chinese people.

Adolescent ; Adult ; Alternative Splicing ; genetics ; Base Sequence ; Child ; China ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Female ; Homozygote ; Humans ; Hyperlipoproteinemia Type II ; blood ; genetics ; pathology ; Lipids ; blood ; Male ; Middle Aged ; Mutation ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Receptors, LDL ; genetics

OBJECTIVETo analyze the clinical characteristics of hospitalized pediatric patients infected with 2009 H1N1 influenza.

METHODSTotally 159 children (83 male and 76 female) with influenza A (H1N1) confirmed by the real-time reverse-transcriptase-polymerase-chain-reaction assay were admitted to a special ward of Capital Institute of Pediatrics from November 2009 to January 2010. Clinical manifestations, laboratory and therapy data from the hospitalized children were collected by designed case report form and analyzed.

RESULTSOut of 159 hospitalized patients, 139 (87.4%) were under the age of 5 years and 34.0% of them had at least one underlying medical conditions. Proportions of the severe cases, pneumonia and underlying medical diseases were similar between the 78 infants and 81 older children. All of these 159 cases had influenza-like symptoms at onset and the most common presentations were fever (115 cases, 72.3%) and cough (154 cases, 96.8%). Five severe cases presented dyspnea, cyanosis and hypoxemia. The virus easily invaded into the lower respiratory tract as indicated by that 61% of the cases had findings consistent with pneumonia by X-ray and/or CT and 21.6% had bacterial co-infection. Part of them had mycoplasma pneumonia (20 cases, 27.0%) or other respiratory viruses (5 cases, 3.1%) co-infection simultaneously. The duration of fever was similar between the H1N1 virus sole infection group and the co-infection group (t = 0.975, P > 0.05), but the average course of the disease and hospitalized days of the latter group were longer than the former (t = 3.182 and 3.190, P < 0.01). The proportion of children with pneumonia in the co-infection group was significantly higher than that in the H1N1 sole-infection group (χ(2) = 4.082, P < 0.05).

CONCLUSIONSMost of the H1N1 infected pediatric patients had mild respiratory symptoms, a few of them developed severe manifestations. Dyspnea and hypoxemia were the early signals for the developing severe cases. Rational and experienced treatment with antibiotics was important addition to the antiviral therapy for those co-infected with bacteria.

Child ; Child, Hospitalized ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; diagnosis ; epidemiology ; pathology ; therapy ; Male

OBJECTIVETo investigate the clinical and pathological features, diagnosis and treatment of cystic lymphangioma of the spermatic cord.

METHODSOne case of cystic lymphangioma of the spermatic cord in a 71-year-old patient was retrospectively analyzed and the relevant literature was reviewed.

RESULTSThe patient, presented with spermatic cord hydrocele, was treated by local excision of the tumor, which was pathologically diagnosed as cystic lymphangioma. No relapse was found during a 3-month follow-up after the operation.

CONCLUSIONLymphangioma of the spermatic cord is a benign tumor. Preoperation ultrasonography and CT are important for determining the location and nature of lymphangioma. Surgical excision is an effective option for the treatment of cystic lymphangioma of the spermatic cord.

Aged ; Genital Neoplasms, Male ; diagnosis ; surgery ; Humans ; Lymphangioma, Cystic ; diagnosis ; surgery ; Male ; Retrospective Studies ; Spermatic Cord ; pathology ; Treatment Outcome

OBJECTIVETo investigate the effect of N-acetyl-cysteine (NAC) and depakine (DP) on the changes of membrane potential and peroxidate in rat cortex neurons exposed to ferrous chloride (FeCl(2)).

METHODSCultured cortex neurons of newly born SD rats were randomly divided into control group (PBS group), model group (FeCl(2) group), NAC pretreatment group (NAC group), DP pretreatment group (DP group) and NAC+DP pretreatment group (NAC+DP group). In the latter three groups, NAC (0.08 mg/ml) and DP (0.1 mg/ml) were added in the cell culture 2 and 3 h before FeCl(2) (1 mmol/L) exposure, respectively. After exposure to FeCl(2), the membrane potential of the neurons was detected with fluorescent dye DiBAC4(3) (bis-(1,3-dibutylbarbituric acid) trimethine oxonol), and the peroxidate level with 2,7-dichlorofluorescin diacetate (H(2)DCF) by laser confocal scanning microscope (LCSM) and nuclear factor-KappaB (NF-KappaB) level with immunocytochemistry.

RESULTSCompared with FeCl(2) group, the expression of NF-KappaB and peroxidate level in the neurons were decreased significantly in NAC and NAC+DP groups (P<0.01), but not in DP group (P>0.05). FeCl(2) depolarized the membrane potential and increased the expression of NF-KappaB in the neurons. Compared with FeCl(2) group, significant changes in the membrane potential were observed in DP and NAC+DP groups (P<0.01) but not in NAC or PBS group (P>0.05).

CONCLUSIONBoth NAC and DP can protect the neurons from FeCl(2)-induced damage but through different pathways, and their combined use can significantly alleviate neuronal damages due to FeCl(2) exposure. Antioxidants such as NAC in combination with antiepileptic drugs may produce favorable effect in prevention and treatment of posttraumatic epilepsy.

Acetylcysteine ; pharmacology ; Animals ; Animals, Newborn ; Cells, Cultured ; Cerebral Cortex ; cytology ; metabolism ; physiopathology ; Female ; Ferrous Compounds ; pharmacology ; Male ; Membrane Potentials ; drug effects ; Neurons ; cytology ; metabolism ; physiology ; Neuroprotective Agents ; pharmacology ; Peroxides ; metabolism ; Rats ; Rats, Sprague-Dawley ; Valproic Acid ; pharmacology

OBJECTIVEThe novel influenza A (H1N1) virus firstly detected in April 2009 in Mexico rapidly spread to many countries including the United States and Canada where humans were infected with the H1N1 virus and deaths were reported. The pandemic virus strain had never been detected in specimen of human beings and swine. It was so highly contagious and widely spread that threatened life of humans globally. This study aimed to analyze clinical data of hospitalized children patients with 2009 novel H1N1 influenza A virus infection confirmed by etiologic tests, and compared with that of seasonal influenza A.

METHODClinical manifestations, laboratory and therapy data from the hospitalized children were collected by designed case report form and analyzed. All patients were enrolled from Capital Institute of Pediatrics from January 2003 to 2010. There were 152 cases in seasonal influenza A group, which was composed of 100 boys and 52 girls. Other 93 boys and 86 girls formed 2009 novel influenza A group.

RESULTInfluenza A was dominate from 2003 to 2008 and the peak season was December and January, while the peak hospitalized time of 2009 novel H1N1 influenza was from November 2009 to January 2010. The median age of seasonal influenza group was 35 months, which was lower than that of novel influenza group (Z = -6.702, P<0.01). Besides, 80.9% of the patients in seasonal influenza group were infants, while the novel influenza A group was mainly composed of infants and pre-school children (chi2 = 40.725, P<0.01). The cases of both groups had influenza-like symptoms at onset and the most common presentations were fever and cough. The duration of fever was much longer in 2009 novel influenza group (Z = -7.173, P<0.01). Patients in two groups nearly had the same symptoms except cough was more frequently presented by novel influenza A group cases (chi2 = 4.109, P<0.05). In laboratory examination, the novel influenza group had more cases with abnormality in blood platelet, CRP, ALT, and CK-MB than that of seasonal influenza group (chi2 = 7.562, 17.245, 4.398, 6.217, P<0.01). Patients in novel influenza A group had more changes in electrocardiogram (chi2 = 24.461, P<0.01). More patients had common underlying medical condition in novel influenza groups than those in seasonal influenza group (chi2 = 12.553, P<0.01). Furthermore, the groups had different age distribution in underlying medical diseases (chi2 = 7.231, P<0.05). Children with 2009 novel H1N1 virus infection tended to catch pneumonia (chi2 = 8.661, P<0.01) and became the severe cases (chi2 = 10.595, P<0.01). They had much higher ICU admission rate (chi2 = 12.873, P<0.01) and longer hospital stay (Z = -2.764, P<0.01).

CONCLUSIONAs a new variant of influenza virus A, 2009 novel H1N1 influenza A had stronger pathogenicity. Children with underlying medical conditions had the high risk to be infected and developed severe manifestations.

Adolescent ; Child ; Child, Hospitalized ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Influenza A Virus, H1N1 Subtype ; Influenza A virus ; Influenza, Human ; epidemiology ; virology ; Male

OBJECTIVETo investigate the effect of nano-TiO(2) intratracheal instillation on the progression of dyslipidemia and atherosclerosis in apolipoprotein E-knockout mice.

METHODSThe nano-TiO(2) was ultrasound with phosphate-buffered saline solutions (PBS) into its suspension for exposure. A total of 46 specific pathogen free (SPF) level of 11-week-old male apolipoprotein E-knockout mice were randomly divided into groups by their body weights: non-treatment group (8 mice), PBS control group (9 mice), high dose group (1.0 mg/ml, 10 mice), medium dose group (0.5 mg/ml, 10 mice), and low dose group (0.1 mg/ml, 9 mice). Except the non-treatment group, mice from other groups were intratracheally instilled with 0.05 ml each time, twice a week. After exposure of 6 weeks, viscera index, blood TC, TG, HDL-C, LDL-C, and organic lipid ratio were assessed as biomarkers. Artery and aortic root issues were assessed by histopathology.

RESULTSAfter 5 weeks exposure, mice body weights in high dose group ((29.7 ± 1.9) g) started to drop, compared to PBS control ((31.3 ± 1.9) g, t = -1.58, P < 0.05) and low dose group ((31.4 ± 1.4) g, t = -1.17, P < 0.05); after 6 weeks, high dose group ((28.8 ± 1.5) g) was lower than PBS control ((30.4 ± 1.9) g, t = -1.60, P < 0.05), non-treatment group ((30.2 ± 1.3) g, t = -1.43, P < 0.05) and low dose group ((30.6 ± 1.0) g, t = -1.83, P < 0.05). TC levels of non-treatment, PBS control, high dose group, medium dose group and low dose group were (2.92 ± 1.18), (3.12 ± 0.73), (4.19 ± 1.86), (3.46 ± 0.72) and (2.57 ± 0.64) mmol/L, respectively; TG levels were (0.39 ± 0.13), (0.39 ± 0.08), (0.60 ± 0.21), (0.55 ± 0.19) and (0.41 ± 0.11) mmol/L, respectively; HDL-C levels were (1.67 ± 0.45), (1.54 ± 0.67), (0.93 ± 0.50), (1.02 ± 0.48) and (1.31 ± 0.64) mmol/L; TG levels of high dose group were higher than that of non-treatment group (t = 1.27, P = 0.03) and low dose group (t = 1.62, P = 0.01); TG levels of medium dose group was higher than PBS control (t = 0.16, P = 0.04), and TC levels of high dose group were higher than PBS control (t = 0.22, P = 0.01), non-treatment group (t = 0.22, P = 0.04) and low dose group (t = 0.20, P = 0.03), and HDL-C levels of high dose group were lower than PBS control (t = -0.61, P = 0.04) and non-treatment group (t = -0.74, P = 0.04); organic lipid ratio of each group were (2.27 ± 0.51)%, (2.06 ± 0.53)%, (2.90 ± 0.50)%, (2.60 ± 0.23)%, (2.24 ± 0.45)%; high dose group were higher than PBS control (t = 0.85, P = 0.00), non-treatment group (t = 0.64, P = 0.03) and low dose group (t = 0.67, P = 0.01); medium dose group was higher than PBS control (t = 0.54, P = 0.02). The plaque lipid content and calcium content which showed the progression of atherosclerosis and plaque rupture were elevated in medium and high dose groups.

CONCLUSIONIntratracheal instillation of nano-TiO(2) can induce dyslipidemia and accelerate the development of atherosclerosis and plaque rupture in ApoE-/-mice.

Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; blood ; chemically induced ; Dyslipidemias ; blood ; chemically induced ; Instillation, Drug ; Lipid Metabolism ; Lipids ; blood ; Male ; Mice ; Mice, Knockout ; Nanoparticles ; Specific Pathogen-Free Organisms ; Titanium ; administration & dosage ; pharmacology