Objective To construct and identify the transgenic plant vector recombinant pBI-Eg95 plasmid of Echinococcus granulosus. Methods Total RNA was extracted from hydatid cyst protoscoleces of Echinococcus granulosus after sonication. A couple of specific primers were designed on the basis of known sequences of Eg95 gene. The desired gene was amplified by PCR technique from the cDNA, and then was cloned into the plant expression vector pBI121 to construct the recombinant pBI-Eg95 plasmid. The recombinant plasmid was electroporated into Agrobocterium tumefaciens (At) LBA4404 strain. The positive recombinant clones were confirmed by restriction endonuclease digestion and characterized by PCR. Results For RT-PCR, a specific band around 471 bp was amplified. The result of DNA sequencing of Eg95 showed the identity with the published sequence. The same band was obtained by restriction endonuclease digestion and PER from the plasmids of positive recombinant At(rAt). Conclusions The recombinant pBI-Eg95 plasmid was successfully constructed, and it provides the basis to further research of the transgenic plant vaccine of Echinococcus granulosus.

Colitis, Ulcerative ; Humans ; Infant, Newborn ; Male

Objective To cultivate and identify the transgenic affalfa containing Echinococcus granulosus Eg95 gene. Methods The alfalfa plants were transformed by co-cultivating alfalfa cotyledons via recombinant Agrobacterium tumefaciens LBA4404 harboring pBI-Eg95. The transgenic alfalfa explants were selected by kanamyein after calli formation, shoots and roots regeneration in the selective medium, the seedlings of transgenic plants were obtained which were finally transplanted into pots containing nutrient soil. After 2-3 months growth, the complete transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene were obtained. To identify the transgenic alfalfa plants, the total DNA, RNA and leaf protein were extracted from fresh leaf tissue of the transgenic alfalfa plants and confirmed by PCR, RT-PCR, SDS-PAGE and Western blot assay. Results A specific band around 471 bp was amplified by PCR with total DNA, and the same band was obtained by RT-PCR with total RNA, which confirmed that the Eg95 gene was stably integrated into the transformed alfalfa genome. SDS-PAGE analysis showed that the relative molecular mass(Mr) of the expressed protein was about 16.5×103, consistent with the Eg95 protein, and the level of Eg95 expression was up to 0.06% of total soluble leaf protein by Bio-Rad Quantity one assay. Western blot verified the expressed protein was reactive with the sera of mice infected with Echinococcus granulosus. Conclusion The transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene are successfully cultivated.

Humans ; Infant, Newborn ; Male ; Polycystic Kidney Diseases ; diagnosis

Objective To compare the single-and double-injection techniques for sciatic nerve block.Methods Sixty ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 18-64 yr,weighing 48-72 kg,undergoing elective unilateral foot and ankle surgery,were randomly divided into 2 groups (n =30 each) using a random number table:single-injection group (group S) and double-injection group (group D).Labat-winnie approach to sciatic nerve block was performed under the guidance of a nerve stimulator.When ankle dorsal or plantar flexion developed,group S received a single injection of 0.5％ ropivacaine 30 ml.When ankle dorsal and plantar flexion developed,0.5％ ropivacaine 15 ml was injected each time in group D.The failure of location was recorded.The time spent performing the procedure,onset time and duration of sensory and motor blockade,and the total time for the block were recorded.The effectiveness of block (success,failure) was assessed.Results The failure rate of location was 10％ in group D.The success rate of block was 93％ in group S,and was 96％ in group D,and there was no significant difference in the success rate of block between the two groups.Compared with group S,the onset time of sensory and motor blockade was significantly shortened,the time spent performing the procedure was prolonged,and no significant change was found in the total time for the block,duration of sensory and motor blockade in group D.Conclusion Single-injection technique is recommended for the sciatic nerve block in the patients undergoing lower extremity surgery.

Bacillus cereus B-04 antagonist to Botrytis cinerea were isolated from samples of tomato soil infected by Botrytis cinerea in Zibo, which are identified through a series of morphological and biochemical characteristics and the sequence of 16SrDNA. Aiming at enhancing the inhibitory effect of this strain, a 4.1kb DNA fragment containing ?-1,3-glucanases gene from pUC1940 was inserted into vector pBE2 and pHY300PLK to construct recombination plasmids, PBE2-glu and pHY300PLK-glu, which were transferred into Bacillus cereus B-04, resulting in a new strain named B-04-glu. Restriction enzyme digestion and ?-1,3-glucanases plate culture confirmed that B-04-glu contained a functional ?-1,3-glucanases gene. Compared to the wild strain B-04, B-04-glu had an increased inhibitory effect against Botrytis cinerea on tomato.

More than ten bacteriophage of E.coli were isolated from the soil and the dung of the fowl-run, then three of named bacteriophage A, C, D which lysis E.coli virulently were selected to investigate biological characterizations. The results showed that high activities were obtained after the phages incubated at 50℃ for 1 h or 60℃ for 30 min. The phages could be alive at the range of pH from 4 to 12, Ca 2+ or Mg 2+ added to the medium could stimulate the lysis of phages. However, the formation of the plaque could be inhibited obviously by adding sodium citrate to the medium.

Abstract: Objective　To investigate the relationship between the body mass index (BMI) levels and the negative conversion time of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid in adult coronavirus disease 2019 (COVID-19) patients and the asymptomatic persons. Methods　Asymptomatic infected patients and confirmed COVID-19 patients admitted to Chengdu Public Health Clinic Center from February 2021 to November 2021 were dynamically included. The epidemiological and clinical characteristics of the objects were collected, and the SARS-CoV-2 nucleic acid testing of the objects during their hospitalization was continuously monitored, and the negative nucleic acid conversion time was recorded. The t test or Wilcoxon rank sum test, χ2 test or Fisher's exact probability method examine were used to distribute characteristics of each group of variables and the connection between different variables, respectively. Then the variables showed differences in distribution (P<0.05) between different BMI groups were included in the multivariate Cox proportional risk regression model. Results　A total of 253 subjects ranged from 18 to 63 years old, with M(P25, P75) age of 37.0 (30.0, 47.0) years old, were included in this study. The male to female ratio was 4.16 to 1. The BMI was (23.97±3.33) kg/m2. 50.59% (128/253) of the objects were overweight or obese, and 78.13% (100/128) were overweight. The negative time of SARS-CoV-2 nucleic acid conversion of all subjects ranged from 1 to 71 days, with M(P25, P75) of 7.0 (2.0, 18.0) days (P<0.001). The negative time of SARS-CoV-2 nucleic acid conversion of the normal weight or the thin, and the overweight or obese were 5.00 (2.00, 19.00) and 8.00 (2.00, 17.75) days respectively. The results of multivariate Cox's proportional hazards regression model showed that the BMI levels may not be associated with the negative conversion time of SARS-CoV-2 nucleic acid (HR=1.090, 95%CI: 0.843-1.410, P=0.510). Conclusions　Adult asymptomatic persons and confirmed COVID-19 patients are mainly middle-aged and young males, and overweight or obesity is relatively common. Overweight or obesity cannot be considered as an independent factor influencing the negative conversion time of SARS-CoV-2 nucleic acid.

0.05);Compared to Gram-negative bacilli group of sputum culture,Gram-positive cocci group had significant diffe-rence in the incidence of gastrointestinal dysfunction and microcirculatory disorders(Pa

Objective To find out the status of drinking-water-borne endemic fluorosis and the effect of preventive measures in Hebei Province,so as to provide a basis to prevent and cure fluorosis.Methods One affected county (city,district) with drinking-water-borne endemic fluorosis was sampled in every city and 10 water improvement projects were investigated in that county.Three villages were taken out in every county.The operating state of the projects,the water fluoride content,and the dental fluorosis of children aged 8-12 and skeletal fluorosis of adults were investigated in these villages.Results A total of 93 projects were investigated,among which 94.6％(88/93) were operating properly and the qualified rate(fluoride content ≤ 1.2 mg/L) of the projects was 54.84％(51/93).Among the 9 large projects,the water fluoride content of 7 projects exceeded 1.2 mg/L.A total of 23 villages with water improvement projects were investigated and only 43.4％ (10/23) of them water fluoride content were lower than 1.2 mg/L.The dental fluorosis rate,the dental fluorosis index and the rate of skeletal fluorosis were 33.2％ (532/1601),0.74 and 5.7％ (66/1155),respectively.In the 7 villages without water improvement projects,2 of them exceeded 2.0 mg/L,but lower than and equal to 4.0 mg/L of water fluorosis and 5 of them were normal(＜ 1.2 mg/L).The dental fluorosis rate,the dental fluorosis index and the rate of skeletal fluorosis were 33.4％ (111/332),0.72 and 2.9％ (10/350),respectively.Conclutions The qualified rate of water improvement projects of fluoride content ≤ 1.2 mg/L is low and the endemic fluorosis is still comparatively serious in Hebei Province.The progress of water improvement projects in the areas with endemic fluorosis should be accelerated and the quality of water improvement projects should be increased.