ABSTRACT
Objective:To provide a potential platform for transferring specific antigen against fish bacterial diseases based on attenuated Lm (EGDe-ΔactA/ΔinlB).Methods: Attenuated Lm (EGDe-ΔactA/ΔinlB) was used to express outer membrane protein K (Ompk),a conserved and effective vaccine candidate in vibrio.The identification of recombinant strains and detection of antigen genes were operated with PCR and RT-PCR,respectively.Results: The results of PCR showed that Lm-Ompk (L-O),Lm-Lmo0576-Ompk (L-L-O) and Lm-P-Ompk (L-P-O) were constructed successfully.The identity of foreign gene was 100％ compared with sequence of NCBI.The analysis of transcription showed that the expressions of Ompk in L-O,L-L-O and L-P-O were significant (P<0.001).Moreover,the expression of Ompk in the condition of antibiotic was higher than that in the BHI without antibiotic (P<0.05).Conclusion: Lm-Ompk (L-O),Lm-Lmo0576-Ompk (L-L-O) and Lm-P-Ompk (L-P-O) were constructed successfully.
ABSTRACT
Soil water is one of the most important components in hydrological cycle. The stable hydrogen and oxygen isotopes in soil water have been increasingly used in the ecological, environment and hydrological research. In view of different techniques for extracting soil water, there is significant difference in theδD andδ18 O composition. This paper presents a method for analyzing hydrogen and oxygen isotopes in soil water by using elemental analyzer and isotope ratio mass spectrometry with accelerated solvent extraction for sample pretreatment. The conditions are: extraction solvent: dichloromethane, temperature: 100 ℃, pressure of 10. 3 MPa, static time:10 min. The samples were extracted three times, and with cycle values of four, four and three, respectively. Comparing with the added water, the deuterium and oxygen isotope values in the extracted soil water enrich 2. 12‰-4. 58‰ and 0. 17‰-0. 93‰, respectively. The reproducibility of replicate extractions of soil water is around ±0. 89‰ for δD and ±0. 37‰ for δ18 O.
ABSTRACT
Objective To investigate the intervention of Amlodipine combined with alanine aminotransferase(ALT)-711 on blood pressure and preventing of target organ remodels of Spontaneously Hypertensive Rats(SHR),and to explore an optimized treatment design.Methods A total of 24 SHRs at 12-Week-old male were randomly divided into 4 groups,with 6 SHRs in each group.In the control group(Group A)the SHR were given normal saline 1 ml/(kg · d)via intragastric administration; the SHRs in Group B were treated with Amlodipine 1 mg/(kg · d) via ntragastric administration; the SHRs in Group C were treated with ALT-711 10 mg/(kg · d)via ntragastric administration;the SHRs were treated with Amlodipine 1 mg/(kg · d)combined with ALT-711 10 mg/(kg · d)via ntragastric administration in Group D.All groups were treated for 8 weeks,and the blood pressure in mouse tail were monitored.After 8 weeks,all SHRs were executed in the condition of anaesthedia.Right cervical artery,heart and kidney were measured through immunohistochemistry,histological stain and imagine analysis.The remodeling of vascular,heart,kidney were compared,and collagen volume fraction,advanced glycation end products (AGEs) and fibronectin (FN) expression were measured.Results (1) After treatment for 8 weeks,blood pressure of SHRs in the 4 groups were (184.72 + 15.14),(108.00 ±10.19),(155.64±10.03)and (99.83 ± 12.27) nun Hg in Group A,B,C and D,respectively.The blood pressure in Group D was the lowest in the 4 groups (F =14.6702,P ＜ 0.05),whereas it is not significantly different from Group B (P ＞ 0.05).(2) The mean thickness of carotid arteries were (0.450 ± 0.023),(0.380 ±0.021),(0.420 ±0.019) and (0.320 ±0.020) mm in Group A,B,C and D,respectively.The mean thickness of carotid arteries in Group D was the lowest in the 4 groups(F =4.1463,P ＜0.05).(3).The mean intimal area of carotid arteries were (19.0 + 1.3) × 105,(25.0 ± 1.4) × 105,(29.8 + 1.5) × 105 and (21.3 ±1.3) × l05 μm2 in Group A,B,C and D,respectively.The mean intimal area of carotid arteries in Group D was significantly smallest than the other 3 groups(F =4.4305,P ＜0.05).(4) The collagen content in different organs in the 4 groups after treatment 8 weeks:in carotid arteries,they were (6.25 ± 0.19)％,(3.56 ± 0.03) ％,(4.25 ± 0.12) ％ and (1.32 ± 0.05) ％ in Group A,B,C and D,respectively.The collagen content in Group D were significantly lower than the other 3 groups (F =4.9316,P ＜ 0.01).In myocardia:collagen content in Group A,B,C and D were (6.02 ± 0.21)％,(4.08 ± 0.14)％,(4.44 ± 0.11)％ and (1.13 ± 0.08) ％ respectively,and it was the lowest in Group D (F =4.2469,P ＜ 0.01).In kidney,the collagen content in Group A,B,C and D were (13.23 ±0.51)％,(7.28 ±0.35)％,(8.33 ±0.22)％ and (5.66 ±0.13)％ respectively,and it was the lowest in Group D(F=8.1038,P ＜0.01).(5).Express rate of AGEs in carotid arteries in Group A,B,C and D were (31.48 ±7.30)％,(20.55 ±5.91)％,(10.48 ±0.98)％ and (7.56 ± 0.68) ％ respectively,with the lowest expression in Group D (F =11.4537,P ＜ 0.01).Express rate of FN in carotid arteries in Group A,B,C and D group were (28.18 ±5.66)％,(17.26 ±5.83)％,(18.20 ±1.44) ％ and (15.12 ± 1.03) ％ respectively,with the lowest expression in Group D (F =9.0036,P ＜ 0.01).Conclusion Amlodipine combined with ALT-711 decreased blood pressure more effectively and prevented the remodel of target organ in SHRs than Amlodipine or ALT-711 alone.