Trichinella spiralis nudix hydrolase (TsNd) gene encoding a 46 kDa protein was expressed in Escherichia coli and the potential of recombinant TsNd protein (rTsNd) as an antigen for the serodiagnosis of trichinellosis was investigated by ELISA and compared with those of ELISA with T. spiralis muscle larval excretory–secretory (ES) antigens. The sensitivity of both ELISA was 100% (30/30), for the detection of anti-Trichinella IgG antibodies in sera of the experimentally infected mice, and the specificity of rTsNd-ELISA and ES-ELISA was 100% (54/54) and 98% (53/54), respectively (P>0.05). Serum anti-Trichinella antibodies were firstly detected by rTsNd-ELISA at 14 days post infection (dpi), then continued to increase with a detection rate of 100% at 36 dpi. The anti-Trichinella antibody levels at different times after infection were statistically different (P<0.05). The results showed that the rTsNd might be a potential candidate antigen for specific serodiagnosis of trichinellosis. But, it needs to be further evaluated with sera of the patients with trichinellosis and other helminthiasis.

Objective To analyze the sensitivity of auxiliary examinations in different periods of sporadic Creutzfeldt-Jakob disease (sCJD).Methods The clinical data of 53 sCJD patients were retrospectively analyzed including the different stages of skull diffusion-weighted magnetic resonance imaging (DWI),24-hour ambulatory electroencephalogram (EEG),18F-FDG PET/CT (PET-CT)and cerebrospinal fluid 14-3-3 protein.When calculating the sensitivity of an auxiliary examination,the diagnostic criteria were defined by combining the specific clinical manifestations with two or more positive results of other auxiliary examinations.Results There were 24,53 and 22 sCJD patients,respectively,met the criterion of early (E),middle (M) and later (L) stage of disease (some patients fit 2 or 3 stages).The sensitivity ofDWl (E:58.3％ M:85.4％,L:94.7％),EEG (E:45.8％,M:62.7％,L:77.8％),14-3-3 protein in cerebrospinal fluid (E:11.1％,M:52.9％) and PET-CT (E:80％,M:100％) increased gradually with disease progression,The sensitivity of PET-CT was higher than the other auxiliary examinations for E and M stages;no PET-CT was conducted in L stage.High signal regions mainly distributed in the cortex in E and M stages,but in L stage,no significant difference was found on the distribution of high signal regions between cortex and basal ganglia.Conclusions The sensitivities of the auxiliary examinations were different for sCJD patients in different stages.Reexaminations in different periods may improve the sensitivity for sCJD diagnosis.The sensitivity of PET-CT was high,and the combination of PET-CT and other auxiliary examinations may play a key role in the diagnosis of sCJD.

Microbial lipase,one of important industrial biocatalysts,has been used widely in many industrial and agricultural fields.It is always the research focus to screen,mine and develop the microbial lipases with novel catalytic activity and high stability.This paper introduces briefly the pathways and methods to mine novel microbial lipase resources from six aspects,including extremophile,metagenome,genome database,protein engineering,immobilization,chemical modification,etc.

To investigate me material basis of Mahuang Fuzi Xixin decoction (MFXD) for anti-inflammation and immune-suppression based on the combined method of serum chemical and serum pharmacological. The LC-MS/MS fingerprints of MFXD, drug-containing serum and blank serum were compared to define the components in plasma. Histamine, β-hexosaminidase released from RBL-2H3 cell infulenced by drug-containing serum at different time points were measured by ELISA. The effect of drug-containing serum on lipopolysaccharide-induced splenocyte proliferation at different time points were determined by MTT. A correlation analysis was made on components of MFXD and pharmacological indexes based the stepwise regression method. After the intragastrical administration with MFXD, 32 components were discovered in rat serum, including 27 prototype components (10 from Mahuang, 13 from Fuzi and four from Xixin) and five unknown components. Compared with blank serum, drug-containing serum could reduce the release of histamine from RBL-2H3 induced by antigen at different time points (P < 0.05); except the 4-hour drug-containing serum, all of the remaining drug-containing serums could inhibit the RBL-2H3 mastocyte degranulation induced by antigen at different time points (P < 0.05). Drug-containing serum could significantly lipopolysaccharide-induced mouse splenocyte proliferation at 15 and 30 min (P < 0.05). A regression analysis was made on the chemical data of components absorbed into blood and pharmacological indexes, i. e. release rate of histamine, release rate of β-hexosaminidase and inhibition rate of splenocyte. This suggested the close correlations among methyl pseudo-ephedrine, pseudoephedrine and histamine released from RBL-2H3 induced by antigen; pseudoephedrine, hypaconine, methyl pseudoephedrine and β-hexosaminidase released from RBL-2H3 induced by antigen; as well as benzoyl hypaconine, benzoylaconine, 14-benzoyl-10-OH-mesaconine, mesaconine and lipopolysaccharide-induced mouse splenocyte proliferation. Methylpseudoephedrine, pseudoephedrine, benzoyl hypaconine, benzoylaconine and mesaconine may be part of material basis of MFXD on anti-inflammation and immune suppression.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacology ; Cell Degranulation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Histamine ; immunology ; Immunosuppressive Agents ; chemistry ; pharmacology ; Male ; Mass Spectrometry ; Mast Cells ; drug effects ; immunology ; Mice ; Rats ; Rats, Wistar ; Serum ; chemistry

OBJECTIVEIn this study, we aimed to investigate the expressions of adhesion molecules on human bronchial epithelial cells and neutrophils in co-culture system, assess the effects of puerarin on suppressing these adhesion molecules expressions, and explore the roles of two crucial signal-transduction elements p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor kappa B (NF-κB) in modulating adhesion molecules expressions.

METHODSNeutrophils and BEAS-2B cells (one human bronchial epithelial cell line) were co-cultured, and adhesion molecules expressions on cell surface were detected using flow cytometry. The mRNA levels of adhesion molecules were assessed by real-time quantitative polymerase chain reaction (real-time qPCR). Phosphorylated p38 MAPK and inhibitor κB were analyzed by Western blot.

RESULTSIn co-culture system, adhesion molecules expressions on BEAS-2B cells and neutrophils were enhanced significantly (P<0.05). Correspondingly, the mRNA levels of adhesion molecules were also increased greatly. Moreover, the pretreatment of peurarin obviously suppressed adhesion molecules expressions on cell surface. Furthermore, phosphorylated p38 MAPK and inhibitor κB in BEAS-2B cells and neutrophils were elevated in co-culture system, but decreased significantly after upon the treatment of peurarin (P<0.05).

CONCLUSIONSCoculture boosted the interactions between human bronchial epithelial cells and neutrophils mimicking airway inflflammation, whereas peurarin decreased the expression of adhesion molecules on cell surface by suppressing the activities of p38 MAPK and NF-κB pathways, and exhibiting its anti-inflflammation activity.

Animals ; Base Sequence ; Bronchi ; cytology ; enzymology ; metabolism ; Cattle ; Cell Adhesion Molecules ; metabolism ; Cell Line ; Coculture Techniques ; DNA Primers ; Down-Regulation ; drug effects ; Epithelial Cells ; enzymology ; metabolism ; Isoflavones ; pharmacology ; NF-kappa B ; metabolism ; Neutrophils ; enzymology ; metabolism ; Phosphorylation ; Real-Time Polymerase Chain Reaction ; p38 Mitogen-Activated Protein Kinases ; metabolism

Objective To explore the clinical outcome of improved technique of Gamma nail in the treatment of intertrochanteric hip fracture of the elderly patients. Methods From March 2002 to October 2006.39 patients with intertrochanteric hip fracture were operated by improved technique of Gam-ma nail.There were 18 males and 21 females at average age of75.7 years(67_98 years).There were 6 patients with type A1 fracture,24 with type A2 fracture and 9 with type A3 fracture according to AO/ASIF classification.Of all.36 patients(92.3%)had osteoporosis.The operation improvements included the following points:(1)The patients were placed at the lateral decubitus position with the fractured limb on the uppermost,with flexion of knee and hip of 60°.The normal hip and knee were flexed as possible.(2)One-off indirect traction reduction was used after general anesthesia. no requirement of continuous mechanical traction.(3)C-arm image intensifier was employed to obtain normal and lateral projections.Results Of all,35 patients were followed up for a mean period of3 years and 2 months, ranging from 6 months to 5 years and 2 months.Operation data showed incision length of(4.3±1.2)cm,mean opera-tion time of(46±10)minutes,intraoperative bleeding volume of(65±26)ml and intraoperative X-ray exposure of(3.0±2.1)times.Postoperative recovery data showed survival in one-year follow up,with ambulation time of(10.5±3.6)days and fracture union time of(10.9±2.1)weeks.Mean Parker's score wag(6.9±3.2)points 6 months after operation. Conclusions Improved technique of Gamma nail can shorten operation duration,reduce operative trauma and bleeding,reduce X-ray exposure and im-prove success rate of surgery.as facilitates early pest-operative recovery and reduces the perioperative mortality rate of the elderly.

A strain was isolated from internal organ of died porcine about 8 weeks with purulent pneumonia,arthritis,pyogenic arthritis and endocarditis in April 2007.Objectives of the study are to confirm the genus of the strain,pathopoiesis,and drug sensitivity.The mainly study methods:the first,the strain was identified by the phenotype and the characteristics of the biochemistry,sequence 16S rDNA genes of the strain was analyzed by molecular biology technology,finally animal experiment and drug sensitivity testing were done.The results of the phenotype and the characteristics of the biochemistry showed that it is greatly similar to Actinomyces hyovaginalis,16S rRNA sequence analysis exhibited the homology achieved to 99.2% com-pared with group III strains of Actinomyces hyovaginalis,and the phyletic evolution analysis also indicated that it has mostly relationship with group III strains of Actinomyces hyovaginalis.Animal experiment dis-covered it has highly pathogenicity to Mus musculus albus;Drug sensitivity testing showed that it is hyper-sensitive to Erycin,Gentamicin and Amikacin.So,the result of the study confirmed that the strain is Actin-omyces hyovaginalis III with the pathogenicity.

Acute Disease ; Fatal Outcome ; Gestational Age ; Humans ; Infant, Newborn ; Male ; Respiratory Distress Syndrome, Newborn ; diagnosis ; therapy

Objective To analyze the clinical features and operative treatment of traumatic cervical spinal fractures in order to improve the prevention and surgical treatment.Methods A review of 324 cases of traumatic cervical spinal fractures admitted to general hospital of Shenyang military area command from January 2006 to December 2015 was performed to analyze their statistical epidemiology.Data included age,gender,etiologies of trauma,anatomical distribution,ASIA classification of neurological deficit and associated injuries.Results The ages of 324 patients ranged from 18 to 85 years,with a mean value of 43.8 years,70.4％ of all the patients (228/324) received operations,the 30 to 39 year-old patients formed the largest group,accounting for about 27.2％ (88/324).Combined cervical and other segments fracture accounting for about 8.0％ (26/324),among which combined cervical and thoracic spine fracture,combined cervical and lumbar spine fracture,combined cervical,thoracic and lumbar spine fracture accounted for 5.9％ (19/324),1.5％ (5/324),0.6％ (2/324) respectively.The frequency of neurological deficit (65.1％),frequency of complications (4.7 ％) was the largest in the ≥ 60 year-old patients group.The frequency of combined injuries(34.2％) was the largest in the 40 to 49 year-old patients group.High fall,road traffic accident and low fall were the most common etiologies,accounting for 34.6 ％ (112/324),32.1％ (104/324) and 18.8 ％ (61/324) respectively.The sex ratio (7.7 ％),frequency of neurological deficit (92.3％) and frequency of combined injuries (34.6％) were the largest in the patients caused by struck by object group.Among 228 patients who received operations,47 patients presented with ASIA A,12 patients with ASIA B,29 patients with ASIA C,63 patients with ASIA D and 77 patients with ASIA E before operation.After operations,46 patients presented with ASIA A,7 patients with ASIA B,23 patients with ASIA C,53 patients with ASIA D and 99 patients with ASIA E.There were 38 patients(16.7％) had got 1 to 2 grade recovery.Conclusion The epidemiological survey of traumatic cervical spinal fractures reveals that the 30 to 39 year-old patients and high fall was the most common age and etiology for injuries of cervical spinal fracture.The ≥60 year-old patients group and struck by object group presented with the highest frequency of neurological deficit,operation can improve the recovery of neurological deficit,the prevention and treatment should be designed according to the injury features of different individuals.

The purpose of this study was to investigate how Zuogui pills from the Kidney-tonifying and Nourishing Yin formula, in combination with the gonadotrophin-releasing hormone antagonist cetrorelix, affected the ovarian local oxidative stress response in decreasing ovarian reserve (DOR) mice. All animal experiments were carried out in accordance with the guidelines and standards established by Jinan University's Experimental Animal Management Committee. Cyclophosphamide (CTX)-treated DOR mice were given Zuogui pills, cetrorelix, or a combination of the two drugs intragastrically. After treatment, there were changes in the estrous cycle, serum sex hormone levels, oxidative stress-related indexes, growth biochemical factor levels, and SIRT1/P53/P21 expression. In comparison to the model group, the Zuogui pills and the cetrorelix+Zuogui pills group had significantly prolonged estrous periods and shortened interestrous periods, and the cetrorelix+Zuogui pills group had a significantly shortened cycle length. Follicle-stimulating hormone (FSH) decreased and estradiol (E₂) increased in all treatment groups compared to the model group, oxidative stress indexes nitric oxide synthase (NOS), nitric oxide (NO), and reactive oxygen species (ROS) decreased, growth biochemical factors brain derived neurotrophic factor (BDNF) and growth differentiation factor 9 (GDF-9) concentrations increased significantly, and leukemia inhibitory factor (LIF) showed no significant change. SIRT1/P53/P21 immunohistochemical results revealed that, when compared to the model group, the expression of SIRT1 increased while the expression of P53 and P21 proteins decreased in all treatment groups, with the cetrorelix+Zuogui pills group having the largest decrease, with significant differences in all indicators. We conclude that cetrorelix combined with Zuogui pills for kidney nourishing and Yin recipe improved the oxidative stress response in the follicle by regulating the SIRT1/P53/P21 pathway, reducing peroxide product production, protecting ovarian function, and regulating ovarian hormone secretion, and its efficacy is superior to that of cetrorelix or Zuogui pills alone.