An extracellar fibrinolytic strain was isolated from fermented shrimp paste. In addition to general physiological and biochemical properties, the strain was identified by 16S rDNA sequence and systematic analysis. The results showed that 16S rDNA sequence of the strain had high similarity with AY601723 and AB195282, suggesting that the strain is a subspecies of Bacillus sp. It was named as Bacillus sp. nov. SK006 by CCTCC. The medium composition and fermentation conditions for fibrinolytic enzyme production were also optimized in the research.

Objective To investigate the mechanism and effect of the spinal cord outlet of the skull base on Chiari Ⅰ malformation with syringomyelia.Methods The cervical spinal cord stem angle (Anbc),slope angle of cervical vertebra (Ansc) of Chiari Ⅰ malformation were measured.In foramen magnum (Llf) and anterior vertebral canal level (Laf),spinal canal(Ac),spinal cord (As) and inferior hernia area (Ah) were measured.Angle,area and ratio were compared in Chiari Ⅰ malformation with syringomyelia,Chiari Ⅰ malformation without syringomyelia and normal control group.Results Ansc,Anbc-Ansc had significant differences among control group and Chiari Ⅰ malformation patients (all P＜0.001).In Llf,Laf,As had significant differences among three groups (all P＜0.05),further comparison of the two showed there were significant differences between Chiari Ⅰ malformation with syringomyelia patients and control group,Chiari Ⅰ malformation without syringomyelia patients and control group in Llf(all P＜0.05).In Llf,Laf,Ac in Chiari Ⅰ malformation with syringomyelia was smaller than control group (P＜0.05).Ah in Llf,Lafand Lh in Llf had no statistical significant difference between Chiari Ⅰ malformation with and without syringomyelia patients (all P＞0.05).In Llf,Laf,As/Ac had statistical significant difference among Chiari Ⅰ malformation with and without syringomyelia patients,control group (all P＜0.001),further comparison of the two showed As/Ac in Llf had statistical significance difference between Chiari Ⅰ malformation with syringomyelia patients and control group (P＜0.05),As/Ac in La had statistical significance difference between Chiari Ⅰ malformation with syringomyelia patients and control group,between Chiari Ⅰ malformation without syringomyelia patients and control group (all P＜0.05),Conclusion The cervical spinal cord,Ansc reducing,narrow vertebral proportion increase are important factors to promote Chiari Ⅰ malformation syringomyelia.

OBJECTIVETo evaluate the effects of angiotensin-converting enzyme inhibitor (ACEI) and angiotensin II type 1 receptor (AT-1 receptor) blocker on the progression of rat pulmonary fibrosis induced by bleomycin A5.

METHODSTwenty-four male Wistar rats were randomized into pulmonary fibrosis model, perindopril treatment, losartan treatment and control groups. In the former 3 groups, pulmonary fibrosis was induced via intratracheal injection of bleomycin A5 (5 mg/kg), after which the rats in the perindopril and losartan groups received intragastric administration of the corresponding agents at the daily dose of 2 mg/kg and 10 m/kg, respectively. The rats in the control group had intratracheal injection of normal saline only. In the 4th week, the histological changes of the lung tissues were examined microscopically with Masson staining. Hydroxyproline content in the lungs was measured, and the protein expressions of AT-1 receptor, TGF-beta1 and IkappaBalpha were examined using Western blotting. DNA binding activity of NF-kappaB was analyzed with electrophoretic gel mobility shift assay (EMSA), and zymography was used to assess the activity of matrix metalloproteinase-2 and 9 (MMP-2, 9).

RESULTSBoth perindopril and losartan treatment significantly reduced the pulmonary fibrosis score, content of hydroxyproline, protein expression of TGF-beta1, DNA binding activity of NF-kappaB and MMP-2, 9 activity, and increased cytoplasmic protein expression of IkappaBalpha. Perindopril treatment lowered the protein level of AT-1 receptor.

CONCLUSIONPerindopril and losartan may inhibit bleomycin A5-induced pulmonary fibrosis in rats by reducing the protein expression of TGF-beta1 and suppressing the DNA binding activity of NF-kappaB and MMP-2, 9 activity.

Angiotensin II Type 1 Receptor Blockers ; therapeutic use ; Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Animals ; Bleomycin ; analogs & derivatives ; Blotting, Western ; Losartan ; therapeutic use ; Male ; NF-kappa B ; metabolism ; Perindopril ; therapeutic use ; Pulmonary Fibrosis ; chemically induced ; drug therapy ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Receptor, Angiotensin, Type 1 ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo investigate the etiology of the outbreak of viral encephalitis in Jinan area in 2003.

METHODSVirus-specific nucleic acid fragments were amplified by random PCR and RT-PCR using specific primers to enterovirus. After sequencing, the gene sequence was handled by the program BLAST for homologous analysis and the software Clustal W 1.82 for multiple sequence alignment to identify the etiology and its genotype.

RESULTSFive strains were isolated from clinical specimens. A gene fragment for one strain was acquired using random PCR, which was highly homologous to enterovirus. Then, the 5' non-translated region and partial VP1 region were amplified and sequenced. The five isolated strains were all identified as Coxsackievirus B5, and what was more, they were most homologous to the strain isolated during the outbreak of aseptic meningitis and encephalitis in Zhejiang province from 2002 to 2004.

CONCLUSIONCoxsackievirus B5 is closely associated with the outbreak of viral encephalitis in Jinan area in 2003. It is an important etiology but other viruses may also played a role which remains to be clarified.

China ; epidemiology ; Disease Outbreaks ; Encephalitis, Viral ; virology ; Enterovirus B, Human ; genetics ; isolation & purification ; Humans ; Polymerase Chain Reaction

OBJECTIVETo explore Chinese medical syndrome distribution features of Japanese encephalitis (JE), and to analyze its correlation between syndromes and features of etiologies and pathogeneses.

METHODSRecruited were 277 patients with confirmative diagnosis of JE from Wuhan Medical Treatment Center, Children's Hospital Affiliated to Chongqing Medical University, Fifth People's Hospital of Guiyang City, Hangzhou Sixth People's Hospital, and Chengdu Hospital of Infectious Diseases between July to September 2012. Chinese medical syndrome distribution features were summarized from their general materials and detailed records of clinical data, including medical history, symptoms and signs, tongue fur, and pulse figures.The frequency of symptoms and signs was calculated according to mild, ordinary, severe, extreme severe degrees. The distribution of Chinese medical syndromes was summarized. And its correlation between syndromes and features of etiologies and pathogeneses were analyzed.

RESULTSAfter clustering analysis, Chinese medical syndromes of JE could be categorized as four groups: toxicity accumulation in Fei and Wei syndrome (TAFWS), brain collateral impaired by poison syndrome (BCIPS), depression of toxicity in the pericardium syndrome (DTPS), exhaustion of yin and yang syndrome (EYYS). BCIPS and DTPS were dominated, accounting for 74.0% (205 cases). The main causes covered evil of summer heat [accounting for 92.42% (256/277 cases)], heat [accounting for 87.73% (243/277 cases)], and toxin [accounting for 99.64% (276/277 cases)].

CONCLUSIONSThe four Chinese medical syndrome types of JE met Chinese medical clinical features of encephalitis. It is induced by infestation of dampness-heat, resulting in toxicity accumulation in Fei and Wei, brain collateral impaired by poison, depression of toxicity in the pericardium. Yin fluid and blood is exhausted as time goes by. Qi and yin are impaired to form intermingled deficiency and excess, and finally causing exhaustion of yin and yang.

Adolescent ; Child ; Child, Preschool ; Encephalitis, Japanese ; diagnosis ; pathology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Medicine, Chinese Traditional ; methods ; Yang Deficiency ; diagnosis ; Yin Deficiency ; diagnosis

OBJECTIVETo investigate the risk factors for pulmonary Candida infection in association with mechanical ventilation and analyze the drug resistance profile of the Candida species that cause the infection.

METHODSA retrospective analysis was conducted 114 patients receiving mechanical ventilation for over 48 h. According to the presence of pulmonary Candida infections, these patients were divided into infected group (n=50, 43.9%) and non-infected group (64 cases). Univariate analysis and multivariate logistic regression analysis were performed to identify the risk factors for the infection, and drug sensitivity test was carried out to evaluate the drug resistance of the Candida species.

RESULTSUnivariate analysis and multivariate logistic regression showed that the presence of at least two underlying diseases (OR=4.758, P=0.009), frequent changes of antibiotics (OR=6.128, P=0.001), and blood albumin below 25 g (OR=15.829, P=0.011) were the independent risk factors for pulmonary Candida infection associated with mechanical ventilation, and prophylactic antifungal treatment (OR=0.062, P=0.012) was a protective factor. Drug sensitivity test showed that Candida albicans was sensitive to most of the antifungal agents (100.0%), but the non-albicans Candida species were resistant to fluconazol (50.0%) and Itraconazole (38.5%).

CONCLUSIONPoor general conditions and frequent changes of antibiotics are the major risk factors for pulmonary Candida infection in patients receiving mechanical ventilation. Drug resistant analysis is helpful in the treatment of the infections.

Adult ; Aged ; Aged, 80 and over ; Candidiasis ; etiology ; China ; epidemiology ; Drug Resistance, Fungal ; Female ; Humans ; Lung Diseases, Fungal ; epidemiology ; etiology ; Male ; Middle Aged ; Pneumonia, Ventilator-Associated ; epidemiology ; microbiology ; Respiration, Artificial ; adverse effects ; Retrospective Studies ; Risk Factors ; Young Adult

Objective: This study aimed to determine the efficacy of a medium-chain triglyceride ketogenic diet on patients with drug-resistant epilepsy over a period of 1 year and 8 months. Methods: Patients with refractory epilepsy on a medium-chain triglyceride ketogenic diet were prospectively enrolled. Their clinical condition and the effectiveness of the ketogenic diet were followed-up every month for 1 year. Adverse events and the reasons for discontinuing the diet were recorded. Results: Fifty-three patients (27 males and 26 females) were enrolled. At the end of the study, 21 patients remained on the diet, 14 of whom were followed-up for 1 year. Among the 53 patients, 22.6% had a more than 50% reduction in seizure frequency, and 16.9% became seizure-free. Conclusions: After a 1-year follow-up, the use of a medium-chain triglyceride ketogenic diet for patients with drug-resistant epilepsy was found to be a safe and effective therapy, and may be considered to bean alternative for patients with difficult-to-control seizures in children as well as young adults.
Epilepsy

A ratiometric fluorescent probe([Eu(Hpfdap)3(CTMR-Phen-N)])was designed and synthesized for detection of singlet oxygen(1O2)with dimethyl anthracyl-containing β-diketone compound 1,1,1,2-pentafluoro-5-(9',10'-dimethyl-2'-anthracene)-3,5-pentanedione(Hpfdap)as recognition ligand and 5-amino-1,10-phenanthroline conjugated with Rhodamine as auxiliary ligand(CTMR-Phen-N).The results of spectral analysis showed that the europium(Ⅲ)complex had almost no fluorescence due to the presence of anthracene ring in the probe.After reaction with 1O2,the anthracene ring was oxidized to endoperoxide,and the fluorescence of the europium(Ⅲ)complex at 614 nm was restored,followed by the increase of quantum yield from 1.60%to 25.16%.The fluorescence intensity of Rhodamine at 575 nm remained unchanged before and after the reaction with 1O2.Therefore,the ratio of fluorescence intensity at 614 nm and 575 nm could be used to measure 1O2 by the developed rationmetric fluorescence method.The probe showed good selectivity for 1O2,and the detection limit(S/N=3)was as low as 3 μmol/L.The fluorescence imaging results demonstrated that the probe had low cytotoxicity and good membrane permeability,and could be applied to fluorescence imaging of 1O2 in Hela cells.

Objective:To investigate impacts of usnic acid(UA)on malignant behavior of gastric cancer cells by regulating the chemokine(C-C motif)ligand 2(CCL2)-CCL2 receptor(CCR2)signal axis.Methods:SGC-7901 cells,a well growing human gastric cancer cell line,were treated with different concentrations of UA,which were grouped into low concentration(UA-L)group(62.5 μmol/L UA),medium concentration(UA-M)group(125 μmol/L UA)and high concentration(UA-H)group(250 μmol/L UA);meantime,the cells were transfected with CCL2 overexpression vector(pc DNA3.1 CCL2),empty vector(pc DNA3.1),silenced CCL2(si CCL2)and negative control(si control),and SGC-7901 cells were treated with 250 μmol/L UA,labeled as UA-H+pc DNA3.1 CCL2 group,UA-H+pc DNA3.1 group,UA-H+si control group and UA-H+si CCL2 group,another untreated SGC-7901 cells were taken as the control group.Flow cytometry,MTT and qRT-PCR were applied to detect cell apoptosis,proliferation,and expres-sion levels of CCL2 and CCR2 mRNA;Western blot was applied to detect expression levels of PD-L1,apoptotic protein(Bax),proli-ferative protein(CyclinD1,CCL2,CCR2)and immune escape related protein(B7H1);after co-culturing with CD8+T cells isolated and cultured in vitro,ELISA was applied to detect levels of IL-4,IFN-γ and IL-10 in the supernatant.Gastric cancer cells in each group were co-cultured with activated peripheral blood mononuclear cell(PBMC)1∶1 for 72 hours,and the sensitivity of gastric can-cer cells in each group to T-cell-mediated killing was compared.Results:Compared with control group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 and B7H1 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-L group,UA-M group and UA-H group were obviously reduced,while apoptosis rate,IL-4 and IFN-γ levels,Bax protein expression were obviously increased(P<0.05);compared with UA-H+pc DNA3.1 group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-H+pc DNA3.1 CCL2 group were obviously increased,while apoptosis rate,IL-4 and IFN-γ levels,and Bax protein ex-pression were obviously reduced(P<0.05);compared with UA-H+si control group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 and B7H1 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-H+si CCL2 group were obviously reduced,while apoptosis rate,IL-4 and IFN-γ levels and Bax protein expression were obviously increased(P<0.05).Conclusion:UA can inhibit gastric cancer cells proliferation,immune escape,and induce apoptosis,which may be related to the inhibition of the CCL2-CCR2 signaling axis.

The aim of the present study is to investigate the expressions of ATP-sensitive K(+) channels (KATP) in pulmonary artery smooth muscle cells (PASMCs) and the relationship with p38 MAPK signal pathway in rats. Male SD rat PASMCs were cultured in vitro, and a model of hypoxia and hypercapnia was reconstructed. PASMCs were divided to normal (N), hypoxia-hypercapnia (H), hypoxia-hypercapnia+DMSO incubation (HD), hypoxia-hypercapnia+SB203580 (inhibitor of p38 MAPK pathway) incubation (HS) and hypoxia-hypercapnia+Anisomycin (agonist of p38 MAPK pathway) incubation (HA) groups. Western blot was used to detect the protein expression of SUR2B and Kir6.1; semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of SUR2B and Kir6.1. The results demonstrated that: (1) Compared with N, H, HD and HS groups, the expressions of Kir6.1 mRNA and protein in PASMCs of HA group were decreased significantly (P < 0.01), but there were no differences among N, H, HD and HS groups (P > 0.05); (2) Compared with N group, the expressions of SUR2B mRNA and protein in H, HD, HS and HA groups were increased significantly (P < 0.05), but there were no differences among H, HD, HS and HA groups (P > 0.05). The results imply that: (1) Hypoxia-hypercapnia, SB203580 didn't change the expressions of Kir6.1 mRNA and protein in PASMCs, but Anisomycin decreased the expressions of Kir6.1 mRNA and protein, so Kir6.1 may be regulated by the other subfamily of MAPK pathway; (2) Hypoxia-hypercapnia raised SUR2B mRNA and protein expressions in PASMCs, but SB203580 and Anisomycin did not affect the changes, so the increasing of SUR2B mRNA and protein induced by hypoxia-hypercapnia may be not depend on p38 MAPK pathway.
Animals ; Anisomycin ; pharmacology ; Cell Hypoxia ; Cells, Cultured ; Hypercapnia ; Imidazoles ; pharmacology ; KATP Channels ; metabolism ; MAP Kinase Signaling System ; Male ; Myocytes, Smooth Muscle ; metabolism ; Pulmonary Artery ; cytology ; Pyridines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfonylurea Receptors ; metabolism ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors