Three purification methods ,based on differential centrifugation,precipitation by polyethylene glycol (PEG)and ultra-speed centrifugation,were compared for purification of an alfalfa mosaic virus (AMVSY)previously isolated from Trifolium repens. The Purified virus was observed under electron microscope, measured,by ultra-violet absorpton analysis and protein determination with SDS-polyacrylamide gel electrophoresis. Our results showed that a method by alternative use of sodium phosphate buffers containing 0. 1mol/L EDTA and 0. 1mol/L MgSO4 achieved the best purification with less miscellaneous protein contamination,integrate virus particles and relatively high yield, which was of 47.6mg virion per hundred grams of fresh leaves of Chanopodium quinoa inoculated with AMV-SY. The coat protein of AMY-SY was tested for about 29 kilo-Dalton.

The 2b protein encoded by Cucumber mosaic virus(CMV)plays an important pathogenicity role in many solanaceous hosts,but mechanism of inducing disease is still unknown.In order to investigate virulence of the 2b protein on Nicotiana glutinosa plants,in terms of chloroplast structure and photosynthesis,a mutant Fny-CMV?2bpro,which cannot express the 2b protein,was achieved by introducing mutant sites in the 2b gene of Fny-CMV.N.glutinosa seedlings were inoculated with wild-type Fny-CMV and the mutant Fny-CMV?2bpro,and were analyzed for symptom expression,chlorophyll content,photosynthetic rate,and ultra-structural alteration of chloroplast.Up to 30 days post inoculation,wild-type Fny-CMV caused symptoms of severe mosaic,leaf deformation,and stunting,reduced photosynthetic rate and chlorophyll content,and altered the ultra-structure and morphological characters of the chloroplasts.However,host seedlings inoculated with the mutant Fny-CMV?2bpro expressed only slight mosaic symptom.Their photosynthetic rates and chlorophyll contents were not significantly different from those of the mock-inoculated plants,and the ultra-structure and morphological characters of their chloroplasts appeared to be normal.The observed low photosynthetic rates and chlorophyll contents were related to the breakage of the chloroplast morphology and ultra-structure.Results of Northern blotting showed that the virulence of 2b protein was associated with high accumulation level of CMV progeny RNAs in systemic leaves.Non-expression of the 2b protein reduced the accumulation levels of its genomic RNAs 1 and 2.The level of subgenomic RNA4,encoding CP protein,was found to be significantly decreased.

Objective The DNA fingerprints of cultivated Pinellia ternata collected from different geographic regions were generated by using AFLP markers to find the feasibility in analyzing their genetic diversity, relationship, and germplasm identification. Methods The DNA polymorphism of 51 cultivated germplasm of P. ternata collected from 17 different habitats and four cultivars of P. pedatisecta (outgroup) were detected by AFLP molecular markers. Results The DNA fingerprints of 51 individuals of P. ternata were obviously distinguished by eight pairs of high polymorphic and efficient primer combinations screened from 64 primer combinations. The phylogenetic clustering results revealed that all the tested cultivars were fully differentiated, and individuals from the same regions were mainly clustered together. Moreover, cultivars from East-China, including Zhejiang and Jiangsu Provinces, displayed clear genetic distinction from other regions. The clustering results were strongly supported by Bootstrap test. Conclusion AFLP Markers can be potentially used in analyzing of genetic diversity, relationship, and germplasm identification of this medicinal plant, and the germplasm from regions of East-China, including Zhejiang and Jiangsu Provinces, displays the relative separate genetic characters from other regions.

Full length cDNAs of Cucumber mosaic virus(CMV)CB7 strain,causing necrosis on Nicotiana glutinosa,were obtained by RT-PCR,using viral genomic RNAs as templates.cDNAs of CMV-CB7 genomic RNAs were cloned and sequenced and results indicated that RNA1,2 and 3 was 3 356 nt,3 045 nt and 2 218 nt,respectively(accordingly Accession Number EF216866,DQ785470 and EF216867).Infectious RNA transcripts from cDNA clones of CMV-CB7 were inoculated onto N.glutinosa and the seedlings of host plants displayed necrosis symptom,whist that of CMV-Fny induced typical mosaic symptoms.Through pseudorecombination between CMV-CB7 and CMV-Fny genomic RNAs,the genetic determinant of necrosis phenotype was mapped to RNA2.Chimeric infectious clones consisting of partial sequences of RNA2 derived from CMV-CB7 and CMV-Fny,respectively,were obtained by Overlapping PCR.Pathogenic analysis with those chimeric RNA2 revealed that 2b gene or 3' UTR of CMV-CB7 RNA2 was responsible for the necrotic pathotype.Northern blotting analysis reflected that both necrotic and non-necrotic viruses accumulated to similar levels of genomic RNAs in host plants.Therefore,necrotic phenotype induced on N.glutinosa was not related to the level of accumulation of CMV genomic RNAs.

Objective: To explore the relationship between the ratio of dietary vitamin A (VitA) to body weight and hypertension among children, so as to provide a reference for blood pressure control through dietary nutritional interventions and childhood hypertension prevention. Methods: Utilizing the baseline survey and followup sample data from the Healthy Children Cohort established in urban and rural areas of Chongqing from 2014 to 2019, structured quantitative dietary questionnaire and selfdesigned questionnaire were used to investigate the information of dietary intake and socioeconomic characteristics of 15 279 children, as well as blood pressure, height, weight measurement. The ratio of dietary VitA to body weight was divided into four groups based on quartiles [≤P25(Q1), >P25~P50(Q2), >P50~P75(Q3), >P75(Q4)]. Generalized linear regression models and Logistic regression models were used to analyze the correlation between ratio of dietary VitA to body weight with blood pressure levels and prevalence of hypertension. Results: The results of the 2014 baseline survey indicated that, after adjusting for confounding factors such as demographic indicators and nutritional intake, significant differences were observed in systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAP) among different groups categorized by the ratio of dietary VitA to body weight (F=157.57, 44.71, 95.92, P<0.01). The baseline ratio of dietary VitA to body weight in children exhibited a negative correlation with DBP, SBP and MAP at baseline and in 2019[baseline: β(95%CI)=-0.65(-0.89--0.42), -0.22(-0.42--0.01), -0.36(-0.56--0.16); 2019: β(95%CI)=-0.77(-1.34--0.19), -0.62(-1.21--0.02), -0.77(-1.34--0.19), P＜0.05]. Compared to Q1 group, the risk of hypertension decreased among children in Q4 at baseline and followup in 2019 [OR(95%CI)=0.63(0.49-0.81), 0.18(0.08-0.42), P<0.01]. Conclusions The ratio of dietary VitA to body weight is significantly negatively correlated with blood pressure levels among children, and dietary VitA deficiency is an independent risk factor for hypertension among children. Measures should be taken to actively adjust childrens dietary nutrition and reduce the risk of childhood hypertension.

OBJECTIVE： To optimize the integrated processing technology for “precise decoction pieces” of Helianthus annuus. METHODS： The contents of total flavonoids and total protein in H. annuus were determined by UV spectrophotometry with rutin and bovine serum albumin as control. Refering to Chinese Pharmacopoeia， the contents of water soluble extract， dilute ethanol extracts and ethyl acetate extract were determined. Based on the different needs such as maintaining quality consistency with the original medicinal materials， preference for anti-gout treatment， preference for liver calming and pain relief， using the contents of total flavonoids， total protein and 3 kinds of polar extract as indexes， gray correlation method was used to optimize the integrated processing technology of 3 kinds of “precise decoction pieces” of H. annuus. RESULTS： Gray correlation analysis showed that the ideal sample sequence of decoction pieces in massive shape dried at 60 ℃ with the original medicinal materials and decoction pieces with preference use of liver calming and pain relief was the most relevant； the ideal sample sequence of ideal sample sequence of decoction pieces in massive shape dried in the shade with decoction pieces with clinical application preference of anti-gout therapy was the most relevant. CONCLUSIONS： Different integrated processing technology for “precise decoction pieces” of H. annuus can be adopted for different needs. If it is necessary to keep the quality consistent with the original medicinal materials or to prepare H. annuus decoction pieces for liver calming and pain relieving， medicinal material can be cut into massive shape and dried at 60 ℃； if it is necessary to prepare H. annuus decoction pieces for anti-gout treatment， cutting into massive shape and drying in the shade can be adopted.

Objective By analyzing the means of donor maintenance from the perspective of intensive care medicine,the difference of clinical indicators before and after donor treatment is obtained,summarizing the treatment experience and effect of clinical maintenance of donor donors,and providing more clinical evidence for the maintenance and treatment of organ donation donors.Methods A retrospective analysis was performed on 76 organ donors collected from the Department of Intensive Care Medicine,Army Medical Center,Chinese People's Liberation Army from January 2014 to December 2021,collected donor maintenance treatment,including mechanical ventilation,continuous renal replacement therapy(CRRT)treatment,enteral nutrition and parenteral nutrition,enteral and parenteral nutrition,plasma exchange,at the same time,the paired t test was used to compare the organ function before and after treatment.Results The lung,liver and kidney of 76 donors were evaluated before and after treatment to determine the number of eligible donors.The difference between the number of eligible donors of kidney and liver before and after treatment was statistically significant(P<0.05).There was no significant difference in lung evaluation(P>0.05).Conclusion Timely and effective organ support therapy for organ transplantation donors is beneficial to the recovery of organ function and improve the utilization rate of organ.