1.Isolation of Bdellovibrio Bacteria from the Gut of Carassius auratus gibelio and the Study of its Biological Characteristics
Microbiology 1992;0(01):-
Bdellovibrio bacteria BDF-H16 was isolated from the gut of Carassius auratus gibelio with Aeromonas sobria.Its shape was ob- served by light microscopy,phase-contrast microscopy,electron microscopy and some of its biological characteristics were also studied.It was demonstrated that BDF-H16 was an gram-negative bacterium and had a bacilliform or arc bacilliform shape with a flagellum at one end.Its size was mostly 0.2?m~0.5?m?0.8?m~1.2?m.It had a wide prey area and could lyse all tested gram-negative bacteria and some gram-positive bacteria.The best lysis conditions to Escherichia coli were 6.75?10~9 cfu/mL of prey bacteria concentration,pH7.0~7.5,28℃.It could grow in the solid culture added 0.85%~5.00% NaCl and was inhibited by enrofloxacin and norfloxacin.
2.Effects of Enrofloxacin on the Growth and Attachment of Bdellovibrio Bacteria
Lu DENG ; Hai-Peng CAO ; Shan HE ; Xian-Le YANG ;
Microbiology 2008;0(08):-
In the experiment, the production of plagues by Bdellovibrio bacteria in solid medium cultivation, the reproduction of Bdellovibrio bacteria in liquid medium cultivation and the attachment of Bdellovibrio bacteria to carrier were observed, which aimed to study the effects of enrofloxacin on the growth and at-tachment ability of Bdellovibrio bacteria BDF-H16. Results indicated that in solid medium cultivation, the production of plagues by Bdellovibrio bacteria BDF-H16 was inhibited by different concentrations (2 ?g/mL, 5 ?g/mL, 10 ?g/mL, 20 ?g/mL, 50 ?g/mL) of enrofloxacin and the inhibitory effects of enrofloxacin became stronger with the increase of the concentration of enrofloxacin. Similarly, in liquid medium cultivation, the reproduction of Bdellovibrio bacteria BDF-H16 was also obviously inhibited by different concentrations ofenrofloxacin and higher concentrations of enrofloxacin such as 10 ?g/mL, 20 ?g/mL, 50 ?g/mL had stronger inhibitory effects on the reproduction of BDF-H16. However, the growth tendency of Bdellovibrio bacteria BDF-H16 was not inhibited in 10 ?g/mL enrofloxacin. Additionally, when zeolite was added, enrofloxacin had also inhibitory effects on the numbers of Bdellovibrio bacteria BDF-H16 attached to zeolite. With the increase of the concentrations of enrofloxacin, the numbers of Bdellovibrio bacteria BDF-H16 attached to zeolite became smaller and smaller. However, the attachment rate of Bdellovibrio bacteria BDF-H16 to zeo-lite became higher under 2 ?g/mL-20 ?g/mL enrofloxacin. The results above showed that enrofloxacin had inhibitory effects on the plague production and reproduction of Bdellovibrio bacteria BDF-H16, but the at-tachment ability of Bdellovibrio bacteria BDF-H16 was strengthened in liquid medium cultivation with 2 ?g/mL-20 ?g/mL enrofloxacin and zeolite, and adding zeolite helped to reduce the adverse effects of en-rofloxacin on Bdellovibrio bacteria BDF-H16.
3.Isolation, Identification and Growth Characteristics of Pseudomonas putida Strain M6 with Malachite Green Decolorization
Yi LI ; Shan HE ; Hai-Peng CAO ; Xian-Le YANG ;
Microbiology 1992;0(01):-
Six bacterial strains with malachite green decolorization ability were isolated from a sediment of aquaculture pond, and strain M6 was selected by further enrichment culture in nutrition broth with malachite green and decolorization rate comparison. The decolorization rate of strain M6 to malachite green was 97.14% in the conditon of 30?C and 150 r/min, and its morphology was observed by gram stain and electronmicroscopy, its physiological and biochemical characteristic was studied by ATB bacteria identification in-strument for identification of bacteria, and its 16S rDNA sequence was determined following PCR amplifi-cation, the sequence was aligned and the phylogenic tree was instructed with those bacterial strains of high identity with strain M6. In addition, its growth characteristics was also studied. The experimental results showed that strain M6 was gram negative and bacilliform with a flagellum at one end. Its size was 0.45 ?m ?0.84 ?m. Its colony produced on common agar plate appeared as round, light blue, dense, hard to choose; 16S rDNA sequence of strain M6 had high identity of 98%~99% with Pseudomonas sp. located in GenBank and strain M6 had the most close relative relation to Pseudomonas putida OW-16 (Locus number: DQ112328.1). Combined the results of the traditional morphological, physiological, biochemical character-istics and 16S rDNA sequence analysis, strain M6 was identified as Pseudomonas putida (Locus number: EU348741.1). Additionally, its growth curve in the condition of 30?C and 150 r/min was as follows: lag phase was 0~4 h, log phase was 4 h~64 h, stationary phase was 64 h~80 h, decline phase was after 80 h. Its best growth conditions were pH 7 and 30?C,and in the rotational speed of 50 r/min to 250 r/min. Its concen-tration increased with the increase in rotational speed.
4.Isolation and Identification of Aeromonas hydrophila Strain X1 from Acipenser baerii and Its Antibiotic Sensitivity
Yuan-Yuan LI ; Hai-Peng CAO ; Shan HE ; Xian-Le YANG ;
Microbiology 2008;0(08):-
A pathogenic bacterial strain X1 was isolated from Siberian sturgeon (Acipenser baerii) suffering with bacterial septicemia. The 50% lethal concentration (LC50) of strain X1 was 5.62?105 CFU/mL, which showed that strain X1 was rather strong virulent to Acipenser baerii. Strain X1 was gram negative and 1.0 ?m~1.2 ?m ? 2.1 ?m~2.4 ?m in size with peritrichous flagella, and had ?-hemolytic activity on rabbit blood agar. By means of ATB expression identification and 16S rDNA sequence analysis, strain X1 was identified as Aeromonas hydrophila (locus number: EU669667), which was the closest relative to Aeromo-nas hydrophila strain ATCC35654 (locus number: X74676.1) with 99% homology. In addition, strain X1 was highly sensitive to cefoperazone and cravit, and intermediately sensitive to ten kinds of antibiotics in-cluding tobramycin, norfloxacin, sulperazone, kanamycin, gentamycin, fortum, vancomycin, neomycina, polymyxin B and lomefloxacin.
5.Study on immunization of Balb/c mice with F1 antigen of Yersinia pestis
He-zhi, LIU ; Xiao-yan, YANG ; Le-le, HU ; Xian-ming, SHI ; Hai-feng, WANG ; Guo-run, DONG ; Yu-gui, LI
Chinese Journal of Endemiology 2009;28(4):369-372
Objective To study the doses and methods of F1 antigen(F1Ag) to immune Balb/c mice during the establishment of hybridoma cell strains. Secreting McAbs against F1Ag of Yersinia pestis. Methods Balb/c mice of seven to nine weeks old were randomly divided into six groups. The first four groups were 150, 100, 50 and 25 μg F1Ag inoculated group, having multipoint hypodermic inoculation of F1Ag of 150, 100, 50 and 25 μg followed by multipoint hypodermic inoculation of F1Ag of 100 μg for a second time and then intraperitoneal injection of 100 μg. Next, hypodermically inoculated group received F1Ag of 100 μg for three times in multiple points. Finally, the intraperitoneal injection group was intraperitoneally inoculated with F1Ag of 100 μg for three times. Emulsification liquid of F1Ag + Complete Frednd's adjuvant(CFA) of equivalence was used in the first inoculation, emulsification liquid of F1Ag + Incomplete Frednd's adjuvant(IFA) balanced mix in the second, F1Ag liquid in the third. One week afterwards, tail blood of the mice was collected to test antibody titers of anti-F1Ag by double antigens sandwich enzyme linked immunosorbent assay (DAgS-ELISA) and trace indirect hemagglutination assay(IHA). Results The levels of antibody of anti-F1Ag in 150,100,50 and 25 μg groups had statistics difference (DAgS-ELISA method: G = 12 173.87,13 440.37,15 024.19 and 4466.72, F= 3.11, P< 0.05;IHA: G = 19 972.32,18 089.40,23 170.47 and 4871.08, F = 4.11, P < 0.05). Immune effect of the 3 groups of 150, 100 and 50 μg was almost the same (P> 0.05), and excelled as compared with that in 25 μg group with statistics difference(DAgS-ELISA method: t = 2.18,2.39,2.73, P < 0.05;IHA: t = 2.54,2.73,3.13, P< 0.05). The titer of F1 antibody had an increasing trend from the 100 μg group to hypodermic group and intraperitoneal injection groups, but without statistics difference (DAgS-ELISA method: G = 8933.44, 9986.16, 13 440.37;IHA: G = 13 777.25,16 384.00, 18 089.40, F = 0.66,0.25, all P > 0.05). Conclusions Hyodermical inoculation of F1Ag with the first dose of 50 μg in multiple points for mouse is appropriate, and a strengthening dose of 100 μg in an intraperitoneal injection may shorten the immune period.
6.Research progress of oral antithrombotic drugs
Li-Ying GONG ; Le-Hong YUAN ; Jie HUANG ; Cheng-Xian GUO ; Yun KUANG ; Wan-Le MA ; Guo-Ping YANG ; Jing-Le LI
The Chinese Journal of Clinical Pharmacology 2016;(3):279-282
Antithrombotic drugs play an important role in the prevention and treatment of thromboembolism , which include anticoagulant , anti-platelet therapies and thrombolytic drugs.In this paper , we review the pharmacological properties of these most commonly used oral antithrom-botic drugs and explore the development of anticoagulant and antiplatelet therapies , in order to guide the safety and rational use of antithrombotic drugs in clinic.
7.Gold-immunochromatography assay for detection of Yersinia pestis F1 antigen
He-zhi, LIU ; Xue-wei, BAI ; Hai-feng, WANG ; Le-le, HU ; Song, ZHOU ; Xiao-yan, YANG ; Guo-yi, DU ; Shun-lin, YANG ; Xian-ming, SHI ; Yu-gui, LI
Chinese Journal of Endemiology 2010;29(6):678-681
Objective To study the sensitivity and specificity of gold-immunochromatography assay (GICA) for detection of Yersiniapestis(Y. pestis ) F1 antigen. Methods Viscera organ(liver and spleen) specimens of 308 mice with virulent Y. pestis infection and 225 control specimens of rats(217 Spermophilus dauricus, 5 mice,3 guinea pigs) were detected by GICA dipstick with monoclonal antibody against plague F1 antigen (F1MAb).Meanwhile, micro-method of reverse indirect hemagglutination assay(RIHA) and bacteria culture were carried out for parallel testing. Results Bacteriological examination of 225 control specimens, and F1 antigen detected with GICA and RIHA were all negative. No cross-reaction with related Yersinia pseudotuberculosis at 1 x 108 cfu/ml level was found in GICA and RIHA. Detection sensitivity of Y. pestis by GICA and RIHA were 2.5 × 105 cfu/ml and 2.0 × 105 cfu/ml, respectively, and of F1 antigen were 1μg/L and 10 μg/L, respectively. Coincidence was 97.94% (522/533) between GICA and bacteriological test, Kappa = 0.959, and the difference was statistically insignificant(x2 = 0.36, P > 0.05); and 97.94%(522/533) between GICA and RIHA, Kappa = 0.959, with statistically significant difference in the positive detection rates(x2 = 9.09, P < 0.05). Out of the 308 infected mice, 284 were positive of plague bacterial cultured, In 284 samples with positive bacterial culture, there were 280 of positive detected by GICA for F1 antigen, positive rate of F1 antigen was 98.59%, higher than that by RIHA[the positive rate of 96.13%(522/533)], with statistically significant difference(x2 = 5.14, P < 0.05). Sensitivity of GICA was 98.59% (280/284), specificity was 97.19% (242/249), positive predictive value (PPV) was 97.56% (280/287),negative predictive value ( NPV ) was 98.37% (242/246), and Youden index was 0.9578. Conclusions GICA is sensitive and specific, fast and simple in detection of F1 antigen of the plague. It's a valuable detection technique for early and rapid diagnosis of plague.
8.Exosome-Transmitted miR-224-5p Promotes Colorectal Cancer Cell Proliferation via Targeting ULK2 in p53-Dependent Manner
Mei Le YANG ; Qi ZHENG ; Jia Xiao LIU ; Xian Xian LI ; Lim VERONICA ; Qi CHEN ; Hua Zhong ZHAO ; Yang Shu WANG
Biomedical and Environmental Sciences 2024;37(1):71-84
Objective To investigate the role and molecular mechanism of exosomal miR-224-5p in colorectal cancer (CRC).Methods The miR-224-5p expression in CRC patient tissues and cell-derived exosomes was measured by laser capture microdissection and qRT-PCR, respectively. Dual-luciferase reporter gene assay was used to determine the target gene of miR-224-5p. The protein expressions of p53 and unc-51 like kinase 2 (ULK2) in CRC cells were detected by western blot. Flow cytometry was used to detect cell cycle and apoptosis. Cell proliferation was measured by CCK8 and EdU assay.Results The miR-224-5p expression was upregulated in CRC tissues and increased progressively with the rise of CRC stage. CRC cells secreted extracellular miR-224-5p mainly in an exosome-dependent manner, and then miR-224-5p could be transferred to surrounding tumor cells to regulate cell proliferation in the form of autocrine or paracrine. Moreover, ULK2 was characterized as a direct target of miR-224-5p and was downregulated in CRC tissues. Interestingly, ULK2 inhibited CRC cell proliferation in a p53-dependent manner. Furthermore, exosome-derived miR-224-5p partially reversed the proliferation regulation of ULK2 on CRC cells.Conclusion Our findings demonstrate that exosome-transmitted miR-224-5p promotes p53-dependent cell proliferation by targeting ULK2 in CRC, which may offer promising targets for CRC prevention and therapy.
9.Causes of orchiectomy: An analysis of 291 cases.
Zhi LONG ; Le-ye HE ; Yu-xin TANG ; Xian-zhen JIANG ; Jin-wei WANG ; Wen-hang CHEN ; Jin TANG ; Yi-chuan ZHANG ; Chi YANG
National Journal of Andrology 2015;21(7):615-618
OBJECTIVETo study the causes of orchiectomy in different age groups.
METHODSWe retrospectively reviewed the clinical data about 291 cases of orchiectomy performed between March 1993 and October 2014 and analyzed the causes of surgery and their distribution in different age groups.
RESULTSThe main causes of orchiectomy were testicular torsion (45.8%), cryptorchidism (32.5%) and testicular tumor (16.9%) in the patients aged 0-25 years, testicular tumor (42.4%), cryptorchidism (25.9%) and tuberculosis (10.6%) in those aged 26-50 years. Prostate cancer was the leading cause in those aged 51-75 years (77.6%) or older (84.0%)), and testicular tumor was another cause in the 51-75 years old men (10.2%). Prostate cancer, testicular tumor, cryptorchidism, and testicular torsion were the first four causes of orchiectomy between 1993 and 2009. From 2010 to 2014, however, testicular tumor rose to the top while prostate cancer dropped to the fourth place.
CONCLUSIONThe causes of orchiectomy vary in different age groups. The proportion of castration for prostate cancer patients significantly reduced in the past five years, which might be attributed to the improvement of comprehensive health care service.
Adolescent ; Adult ; Age Factors ; Aged ; Causality ; Child ; Child, Preschool ; Cryptorchidism ; surgery ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Orchiectomy ; statistics & numerical data ; Prostatic Neoplasms ; surgery ; Retrospective Studies ; Spermatic Cord Torsion ; surgery ; Testicular Neoplasms ; surgery ; Tuberculosis, Male Genital ; surgery ; Young Adult
10.Transverse preputial island flap technique (Duckett's procedure) for hypospadias repair: a report of 356 cases.
Xian-Zhen JIANG ; Jian-Fu YANG ; Qing ZENG ; Bo WAN ; Le-Ye HE
National Journal of Andrology 2011;17(7):622-624
OBJECTIVETo evaluate the effect of the transverse preputial island flap technique (Duckett's procedure) for hypospadias repair.
METHODSA total of 356 patients with hypospadias were treated by Duckett's procedure from March 1995 to December 2010, of whom 324 (91.0%) were younger than 14 years. The length of urethra repair ranged from 1.5 to 10 cm.
RESULTSThe total success rate of Duckett's procedure was 91.0%. Urethra fistula occurred in 30 cases, external orifice stricture in 1, and urethral anastomosis stricture in another. There were no significant differences in the rate of complications among either different age groups or different surgical times (P > 0.05).
CONCLUSIONDuckett's procedure remains the first choice for the one-stage repair of hypospadias, especially applicable to hypospadias with chordee.
Adolescent ; Adult ; Child ; Child, Preschool ; Foreskin ; surgery ; Humans ; Hypospadias ; surgery ; Infant ; Male ; Surgical Flaps ; Young Adult