Twenty-nine antagonistic bacillus strains, isolated from some Chinese traditional medicine and fermented food , inhibit the growth of Fusarium oxysporum f. sp. Vasinfectum and Verticillium dahliae Kleb. And twelve of them are able to produce antagonistic proteins. Among the twelve strains, five (H110, H184, H216, B316 and B382) showed higher antibacterial activity. Furthermore, H110 and H184 were identified as Bacillus subtilis, and H216, B316 and B382 as Bacillus licheniformis based on physiological and biochemistry experiments. The antagonistic proteins of five strains were all thermostable, resistant to proteinase K and trypsin, while H184 and H216 partially sensitive to pepsin.

Objective To evaluate the clinical application of serum CYFRA21-1 detection during the diagnosis of lung cancer. Methods Serum level of CYFRA21-1 in 196 patients with lung cancer and 39 healthy controls were determined by the Elctro-chemiluminescence Immuno-assay ( ECLIA) , respectively. Results Serum level of CYFRA21 -1 ( range from 0. 30 μgg/L to 500.00 μg/L) in patients with lung cancer were significantly higher than those in the healthy control ( range from 0. 27 μg/L to 3. 29 μg/L,Z = -4. 65,P < 0.01). ROC analysis of diagnosis efficiency showed that the area under curve ( AUC) was 0. 735 (P < 0. 001 ). The confidence interval of 95% was from 0. 663 to 0. 807. Positive detection rate of CYFRA21-1 was 48. 8% (39/80) in squamous,31.7% (20/63) in adenocarcinoma and 22. 6% (12/53) in small cell lung cancer (SCLC) respectively, as well as total positive rate was 36. 2% (71/196). Stepwise regression analysis showed that serum level of CYFRA21-1 had a linear correlation with the squamous and the metastasis to liver or bone (r =0. 505 ,P <0. 05). Conclusions Serum level of CYFRA21-1 increases the diagnostic sensitivity, which is helpful for determining histological types, monitoring the metastasis of liver and bone, and evaluating the treatment response in patients with lung cancer.

To remedy the limitations of traditional numerical classification softwares,a new application,X-Cluster,was developed by using various design patterns.X-Cluster had powerful functions to support the researching of numerical classification,and testified by some classify studying about Bacillus spp..

BACKGROUNDAdvances in catheter ablation procedures for the treatment of supraventricular arrhythmias have created the need to understand better the morphological and electrophysiological characteristics of the inferior nodal extension (INE) and transitional cellular band (TCB) in the atrioventricular (AV) junctional area.

METHODSFirstly, we observed the histological features of 10 rabbit AV junctional areas by serial sections under light microscopy. Then we recorded the action potentials (APs) of transitional cells (TCs) in the INE, TCBs, AV node, and ordinary right atrial myocytes from the AV junctional area of 30 rabbits using standard intracellular microeletrode techniques.

RESULTSUnder light microscopy, the INE appeared to be mostly composed of transitional cells linking upward to the AV node. Four smaller TCBs originated in the orifice of the coronary sinus, the region between the septal leaflet of the tricuspid valve and the coronary sinus, the inferior wall of the left atrium, and the superior interatrial septum, respectively, all linking to the INE or the AV node. Compared with ordinary atrial myocytes, the AP of the TCs in both the INE and the TCBs had a spontaneous phase 4 depolarization (not present in ordinary atrial myocytes), with a less negative maximum diastolic potential, a smaller amplitude, a slower maximum velocity of AP upstroke, and a longer action potential duration at 50% repolarization (APD50) and at 30% repolarization (APD30). The AP characteristics of these TCs were similar to those of the AV node, except that the velocities of the phase 4 spontaneous depolarization were slower and their action potential durations at 90% repolarization (APD90) were shorter. Moreover, APD50 and APD30 of the TCs of the TCBs were shorter than in the case of TCs of the AV node.

CONCLUSIONSThe TCs of the INE and TCBs are similar to slow response automatic cells. They provide a substrate for slow pathway conduction. In addition, repolarization heterogeneity exists in the AV junctional area.

Action Potentials ; Animals ; Atrioventricular Node ; cytology ; physiology ; Female ; Male ; Rabbits

OBJECTIVETo develop a real-time quantitative polymerase chain reaction(PCR) assay for detecting Rickettsia rickettsii.

METHODSThe primers and TaqMan-MGB probe were designed according to the ompB gene of R. rickettsii. A DNA fragment of ompB gene amplified from R. rickettsii by PCR was used as a standard template for the development of the method.

RESULTS5 copies of ompB fragments of R. rickettsii were detected. The genomic DNA of R. rickettsii was detected by the developed quantitative PCR assay. However, the genomic DNA from another rickettsial or bacterial agent was not determined. Through this developed method, the positive results were obtained from the animals and cells, artificially infected with R. rickettsii.

CONCLUSIONThe real-time quantitative PCR assay seemed to be highly sensitive and specific which might be used to rapidly detect R. rickettsia DNA in various samples and to early diagnose patients infected by R. rickettsii.

DNA Primers ; Humans ; Polymerase Chain Reaction ; methods ; Rickettsia rickettsii ; genetics ; Rocky Mountain Spotted Fever ; diagnosis ; Sensitivity and Specificity

OBJECTIVETo develop a quantitative real-time polymerase chain reaction (PCR) for detecting Bartonella henselae.

METHODSAccording to the 16S-23S rRNA intervening sequences (IVS) specific for B. henselae, one pair of primers and one TaqMan-MGB probe were designed. A quantitative real-time PCR was developed with the primers, the probe, and the IVS, a standard template, in DNA sequence detection system (ABI 7900HT).

RESULTSThe standard curve was established with the standard template and the relationship between the value of threshold cycle (Ct) and the DNA copy number was linear (r = 0.997). The sensitivity of this quantitative real-time PCR was about 1000 times higher than that of a common PCR used to detect homologous DNA. By this quantitative real-time PCR, the DNA sample of B. henselae was positively detected but not from other rickettsial or bacterial DNA samples. The variation coefficients of intra- and inter-assay reproducibility were 0.2%-1.9%. Using the real-time quantitative PCR to detect samples from mice that were experimentally infected with B. henselae, the small amount of B. henselae DNA was detected in blood samples on days 2, 3, and 5 and large amount of B. henselae DNA was detected in spleen samples on days 1 and 2 after infection.

CONCLUSIONResults from our study suggested that this quantitative real-time PCR was highly specific, sensitive and with good repeatability for detection of B. henselae. It seemed quite useful for rapid detection of tiny DNA of B. henselae in various samples and laboratory diagnosis of bartonellosis caused by B. henselae.

Animals ; Bartonella Infections ; diagnosis ; Bartonella henselae ; genetics ; DNA, Bacterial ; analysis ; Mice ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity

Sexual reproduction is marked by the fusion of the sperm cell with the oocyte during fertilization to produce the diploid zygote, in which the lipids in the sperm plasma membrane play an important role. Due to the loss of most cell organelles and DNA transcription, spermatozoa lack protein expression and vesicular transport. However, the lipids of the sperm plasma membrane undergo complicated dynamic changes, which may facilitate the capacitation, binding with zona pellucida, acrosome reaction and fusion of the sperm cell with the oocyte. This paper summarizes the progress in the studies of the lipids in the sperm plasma membrane, their composition, structure, peroxidation, metabolism and role in fertilization.
Acrosome Reaction ; Animals ; Cell Membrane ; chemistry ; Fertilization ; Humans ; Male ; Membrane Lipids ; metabolism ; Sperm Capacitation ; Spermatozoa ; chemistry

Objective To observe the pathologically regressive changes of rat models of Parkinson's disease (PD) induced by Lactacystin (Lac), and to investigate the pathogenesis of PD.Methods Totally 32 SD rats were randomly divided into 2 groups: test group and control group. Lac, a selective proteasome inhibitor, was unilaterally injected stereotaxically into the left substantia nigral pars compacta (SNc) of rats. Equal volume of saline was injected in control group. On day 1, 3, 5, 7, 9, 11, 14 and 21 post-in jection, we observed the behavioral changes of the rats and pathological changes in substantial nigra and striatum, and detected the expressions of tyrosine hydroxylase (TH) and α-synuclein by immunohistochemistry (IHC) and HE staining and the expressions of their mRNA by RT-PCR. Results Behavioral changes were found in the rats of test group. By HE staining, after Lac-treatment, microglia was increased on the 1st day (3501.92±57.32), 4895.50±52.67 on day 7, 5340.18±52.87 on day 21 (P<0.05 vs control group 3271.23±63.76). By IHC, after Lac-treatment, the test group manifested retrograde dopaminergic neuron degeneration in nigra; the number of neurons was 568.57±36.39 on day7 and 119.67±21.06 on day 21 (P<0.05 vs control group 679.76±30.24). By IHC, after Lac-treatment,expression of TH-positive nerve fiber in left striatum was decreased on day 7, being 0.1953±0.0076 (P>0.05 vs control group 0.2412±0.0067); it was 0.0781±0.0013 on day 21, significantly different from control group 0.2412±0.0067 (P<0.05). Meanwhile, in the test group, the detection of mRNA expression displayed that TH neurons were reduced, while α-synuclein mRNA was accumulating in survival TH neurons. Conclusion Pathological changes in PD models induced by lactacystin are retrograde along with time going on, and consistent with the characters of delitescence and slow progressions in PD.

BACKGROUNDFew studies have explored the inward sodium current (INa) kinetics of transitional cardiomyocytes. This study aimed to explore the kinetics of transitional cardiomyocytes types alpha and beta.

METHODSThe whole-cell patch clamp technique was used to study the rapid INa of isolated transitional cardiomyocytes in the Koch triangle of rabbit hearts.

RESULTSMaximal amplitude and density of INa in type alpha and type beta was (-1627 +/- 288) pA (alpha), (-35.17 +/- 6.56) pA/pF (beta) and (-3845 +/- 467) pA (alpha), (-65.64 +/- 10.23) pA/pF (beta) (P < 0.05). Steady state activation curves of INa, fitted to a Boltzmann distribution for both types, were sigmoid in shape. Half activation voltage and slope factors did not significantly differ between types at (-43.46 +/- 0.85) mV (alpha), (-41.39 +/- 0.47) mV (beta) or (9.04 +/- 0.66) mV (alpha), (11.08 +/- 0.89) mV (beta). Steady state inactivation curves of INa, fitted to a Boltzmann distribution in both types were inverse "S" shape. Half inactivation voltage and slope factors were (-109.9 +/- 0.62) mV (alpha), (-107.5 +/- 0.49) mV (beta) and (11.78 +/- 0.36) mV (alpha), (11.57 +/- 0.27) mV(beta), (P > 0.05), but time constants of inactivation were significantly different at (1.10 +/- 0.19) mV (alpha) and (2.37 +/- 0.33) ms (beta), (P < 0.05). Time constants of recovery from inactivation of INa for both types were (122.16 +/- 27.43) mV (alpha) and (103.84 +/- 28.97) ms (beta) (P < 0.05).

CONCLUSIONSTransitional cardiomyocytes in rabbit hearts show a heterogeneous, voltage gated and time dependent fast inward sodium current. Types alpha and beta show the features of INa similar to those in slow- and fast-response myocytes, with probably better automaticity and conductivity, respectively.

Animals ; Female ; Ion Channel Gating ; Kinetics ; Male ; Membrane Potentials ; Myocytes, Cardiac ; metabolism ; Rabbits ; Sodium Channels ; physiology

OBJECTIVETo develop a quantitative real-time polymerase chain reaction (PCR) for detecting Rickettsia prowazekii.

METHODSPrimers and TaqMan-MGB probes designed based on ompB gene of R. prowazekii, were used to develop this method.

RESULTSFor the quantitative real-time PCR, the relationship between the values of threshold cycle (Ct) and the DNA copy number was linear (r = 0.999) and the sensitivity was about 100 times higher than that of the nested PCR for detecting the same DNA sample. The results of the genomic DNA samples of other rickettsial and bacterial agents detected by real-time PCR were all negative. DNAs extracted from blood samples of guinea pig infected with R. prowazekii were examined by real-time PCR and the positive results were obtained from some of these samples. However, the results of some samples in nested PCR assay were all negative.

CONCLUSIONThese results suggested that the real-time PCR was highly specific and sensitive for detection of R. prowazekii that was useful for the detection of tiny DNA of R. prowazekii in blood samples from patients suspected of having epidemic typhus.

DNA Primers ; DNA, Bacterial ; analysis ; Humans ; Polymerase Chain Reaction ; methods ; Rickettsia prowazekii ; genetics ; isolation & purification ; Sensitivity and Specificity ; Typhus, Epidemic Louse-Borne ; diagnosis