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 To investigate the infection rates of mycoplasma penetrans (Mpe),mycoplasma pneumoniae (Mp) and mycoplasma ferments (Mf) in patients with IgA nephropathy (IgAN),chronic kideny disease (CKD),and heathy people,to compare the difference of infection rate,and to analyze the association of mycoplasma infection and clinicopathological features in IgAN.Methods Blood samples were collected from 118 patients in IgAN group,90 patients in CKD group,and 89 cases in health control group.DNA of Mpe,Mp and Mf was detected in plasma by PCR.Positive cases were confirmed by Southern blot.According to mycoplasma infection,IgAN patients were divided into two groups,then analyzed the clinicopatholgical features.Results (1)Genus,Mpe,Mp,and Mf positive rates were 33.05％,16.1％,25.45 ％,and 8.47％ in IgAN group,respectively; 5.56％,2.22％,5.56％,and 2.22％ in CKD group,respectively; and 3.33 ％,1.11％,2.22％,and 0 in health group,respectively.Compared with CKD and health group,patients in IgAN group had a higher infection rate in Genus,Mpe,and Mp (P ＜ 0.05).In IgAN group,10 patients had three kinds of mycoplasmas infection at the same time,and positive rate was 8.47％ much higher than CKD group (positive rate was 2.22％) (P ＜ 0.05).(2) Based on mycoplasm detection results,IgAN patients were divided into two groups,overlapping infection group and mycoplasma negative group.In overlapping infection group,the mean age of onset was much younger than negative group.Compared with negative group,overlapping infection group had higher tonsillitis and urinary tract infection rate,more severe microscopic hematuria and tubulointerstitium lesion (P ＜ 0.05).Conclusions Patients with IgAN had higher infection rate of Genus,Mpe and Mp,compared with CKD patients and health people.Compared with mycoplasma negative group in IgAN patients,more severe microscopic hematuria and tubulointerstitium lesion in overlapping infection group,which suggested that infection of Mpe might have some possible connection with IgAN.

[Objective] To investigate the effects of the bioactive constituents of green tea polyphenols epigallocatechin gallate (EGCG) on low density lipoprotein receptor (LDLR) function of HepG2 cells.[Methods] The optimal concentration and cell proliferation of HepG2 cells were determined by CCK8 assay,and Western blotting was used to determine LDLR and PCSK9 protein levels,respectively,and LDL uptake in HepG2 cells was detected by fluorescence microscope.[Results] EGCG elevated LDLR protein expression,reduced PCSK9 protein expression and promoted LDL uptake in HepG2 cells.[Conclusion] EGCG may increase LDLR abundance by down-regulating PCSK9 protein and attenuating LDLR protein degradation,which providing a new approach for lipid lowering therapy.

Objective To observe the gastrointestinal function influence in the compound catechu antidiarrheal ointment when acted on normal and diarrheic mice .Methods The antidiarrheal function of the compound catechu antidiarrheal ointment was ob-served through the mice small intestinal propulsion experiment ,gastric emptying experiment and mice diarrhea situation influence test of senna .Results This ointment could reduce the mice small intestine propulsion rate which significantly restrained the mice small intestine propulsion ;Also it could improve the residual rate of mice stomach contents ,significantly restrained their gastric emptying velocity .Conclusion The compound catechu antidiarrheal ointment has strong and lasting effect on stopping diarrhea .

Objective To observe the effects on and the possible mechanism of early exercise training underlying the mobilization of circulating endothelial progenitor cells in patients with acute ischemic stroke.Methods One hundred and twenty patients with acute ischemic stroke were randomized into two groups:an early exercise group (treated with exercise training,n =60) and a control group(no exercise training,n =60).Meanwhile,each group was divided into two different age groups (50-68 years group of 32 cases,＞ 68 years group of 28 cases in exercise group;50-68 years group of 29 cases,＞ 68 years of 31 cases in control group).The amount of circulating endothelial progenitor cells in peripheral blood was accounted by flow cytometry (FCM),while the level of vascular endothelial cell growth factor (VEGF) in blood serum was examined by enzyme-linked immunoassay (ELISA).National Institutes of Health Stroke Scale (NIHSS),Fugl-Meyer Assessment (FMA) and modified Barthel index (MBI) were used to evaluate the patients at 1st day and 14th day after exercise.Results The amount of circulating progenitor cells after 14 days of exercise training in exercise group (from 27.93 ± 6.08/ml to 457.49 ± 73.02/ml)is higher than in control group(from 28.29 ± 5.93/ml to 81.87 ± 9.92/ml) (P ＜ 0.01).Compared with the control group,the level of VEGF at 14th day of exercise group is significantly higher (P ＜ 0.01).The score of NIHSS,FMA,MBI were not significantly different between exercise group and control group (P ＜ 0.01).However,the score of NIHSS was decrease in both exercise group and control group after treatment.Furthermore,the amount of circulating progenitor cells was not significantly different between the age groups either in exercise group or control group (P ＞ 0.05).Conclusion Early Exercise training may promote the mobilization of the circulating progenitor cells,which might be related to the increase of VEGF.The mobilization of the EPCs has no effect on the early neurological function after acute ischemic stroke.Age is not associated with the amount of circulating endothelial progenitor cells.

Objective To observe the effect of hyperbaric oxygen (HBO)on neuropathic pain and detect the expression of Wnt/β-catenin signaling pathway in the mechanism of treatment. Methods A total of 30 healthy male SD rats were randomly divided into three groups (n =10):group S(sham group),group CCI(chronic sciatic nerve constrictive injury group),group HBO(HBO treat-ment for seven days group).Rats of group HBO received HBO one time per day for 7 days since the 1st day after surgery.Rats of group S and group CCI were just placed inside the chamber for approxi-mately 100 min,no HBO treatment.The mechanical withdrawal threshold (MWT)and thermal with-drawal latency (TWL)of all rats were measured on 1 d before CCI,1 d,3 d,5 d,7 d after CCI respec-tively.The animals were sacrificed after the last measurement of pain threshold,and the lumbar spinal cord specimen were taken to detect the expression of Wnt3a,β-catenin by immuno-histochemis-try and Western blot analysis.Results Compared with group S,MWT decreased significantly and TWL shortened significantly in CCT and HBO groups at 1,3,5,7 d after operation (P <0.05).Com-pared with group CCI,MWT was elevated and TWL was prolonged in group HBO at 1,3,5,7 d after operation (P <0.05).The Wnt3a,β-catenin expression increased after CCI treatment compared with group S and group HBO (P <0.05).Compared with group CCI,the expression of Wnt3a,β-catenin decreased significantly in the group HBO and group S at 7 d after operation (P <0.05).There was no significant difference between group S and group HBO.Conclusion HBO post-conditioning may relieve the neuropathic pain of rats by inhibiting the activation of spinal path of Wnt3a,β-catenin.

OBJECTIVETo investigate the relationship between the 3 polymorphic loci of endothelin receptor type A (EDNRA) gene and intracranial aneurysms.

METHODSThree EDNRA gene polymorphisms (rs5335, rs6842241, and rs6841581) were genotyped using polymerase chain reaction-restriction fragment length polymorphism analysis. The genotype and allele frequencies were calculated in the patients and the control group to analyze the association between the gene and the size of aneurysms.

RESULTSNo significant difference was found in the distribution of the EDNRA gene genotypes or allele frequencies between the patients and the control subjects. Only GG genotype of rs6841581 was found to significantly correlate with the size of aneurysms.

CONCLUSIONEDNRA gene rs6841581 has significant associations with the size of intracranial aneurysms, indicating a possible role of EDNRA in the genetic mechanisms of intracranial aneurysms and subarachnoid hemorrhage.

Adult ; Aged ; Case-Control Studies ; Female ; Gene Frequency ; Genotype ; Humans ; Intracranial Aneurysm ; genetics ; pathology ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Receptor, Endothelin A ; genetics

Methyl (S)-4-(6-amino-9H-purin-9-yl)-2-hydroxybutanoate (DZ2002) is a potent reversible inhibitor of S-adenosyl-L-homocysteine hydrolase (SAHH). Due to its ester structure, DZ2002 is rapidly hydrolyzed in rat blood to 4-(6-amino-9H-purin-9-yl)-2-hydroxybutyric acid (DZA) during and after blood sampling from rats; this hampers accurate determination of the circulating DZ2002 and its acid metabolite DZA in rats. To this end, a method for determining the blood concentrations of DZ2002 and DZA in rats was developed by using methanol to immediately deactivate blood carboxylesterases during sampling. The newly developed bioanalytical assay possessed favorable accuracy and precision with lower limit of quantification of 31 nM for DZ2002 and DZA. This validated assay was applied to a rat pharmacokinetic study of DZ2002. After oral administration, DZ2002 was found to be extensively converted into DZA. The level of systemic exposure to DZ2002 was significantly lower than that of DZA. The apparent oral bioavailability of DZ2002 was 90％–159％. The mean terminal half-lives of DZ2002 and DZA were 0.3–0.9 and 1.3–5.1 h, respectively. The sample preparation method illustrated here may be adopted for de-termination of other circulating ester drugs and their acid metabolites in rodents.

Objective:To analyze the epidemiological and clinical characteristics of patients with tsutsugamushi disease in Hainan Province, so as to provide basis for diagnosis, treatment, prevention and control of the disease.Methods:Clinical data of 93 patients with tsutsugamushi disease in the Second Affiliated Hospital of Hainan Medical University from January 2018 to December 2019 were collected. Epidemiological data, clinical manifestations, examination results, treatment and outcome of the patients were retrospectively analyzed.Results:Of totally 93 patients with tsutsugamushi disease, 48 were male and 45 were female, with an average age of 50.73 years old. The peak time of onset was from June to October, with 46 cases (49.46%). Seventy cases (75.27%) were farmers, and 84 cases (90.32%) had a clear history of field work or grassland contact before onset. The clinical manifestations were fever (93/93, 100.00%), eschar or ulcer (42/93, 45.16%), headache (65/93, 69.89%), chills (60/93, 64.52%), rash (35/93, 37.63%), lymphadenopathy (51/93, 54.84%) and fatigue (40/93, 43.01%). Laboratory examination results: eosinophil reduced (74/93, 79.57%), platelet reduced (32/93, 34.41%); alanine aminotransferase, C reactive protein (CRP), procalcitonin (PCT) and erythrocyte sedimentation rate (ESR) were increased in 81.72% (76/93), 97.85% (91/93), 20.43% (19/93) and 72.04% (67/93) of the patients, respectively. Eighty-four cases (90.32%) had abnormal results of chest imaging. All patients were cured by doxycycline.Conclusions:Tsutsugamushi disease is prevalent in summer and autumn in Hainan. It has various clinical manifestations. Doxycycline is an effective treatment drug. It is suggested that the relevant departments should strengthen the training of clinical diagnosis and treatment of tsutsugamushi disease by local clinicians to reduce the misdiagnosis rate.

Objective: To analyze the status of dietary sodium intake of Chinese adult residents in fifteen provinces in 2015. Methods: Data were collected from 2015 China Nutrition Transition Cohort Study. This cohort study used a three consecutive 24-h retrospective method to record food intake and weighing method to record household condiment consumption. In this study, 14 847 residents aged 18 years and over with complete data from 15 provinces in 2015 were selected for the comparison of dietary sodium intake (median) with different characteristics. Results: Among the 14 847 residents, 47.1% were males, and the rural and northern residents accounted for 60.1% and 37.7%, respectively. Median sodium intake was 3 960.0 mg/d among residents in 2015. The sodium intake of males (4 272.1 mg/d) was higher than that of females (3 716.6 mg/d). Across age subgroups, sodium intake was highest among residents aged 45-59 years (4 257.0 mg/d; 18-44 years old, 3 867.1 mg/d; ≥60 years old, 3 799.0 mg/d). Sodium intake was higher in rural area (4 042.9 mg/d) than in urban area (3 866.8 mg/d), higher in north (4 229.2 mg/d) than in south (3 806.8 mg/d) (all P values <0.05). 11.8% of residents with dietary sodium intake were below 2 000 mg/d. The main sources of dietary sodium were condiments (79.4%), regular cooking foods (11.9%) and processed foods (8.7%). Among the condiments, the salt and soy sauce accounted for 63.6% and 10.4%, respectively. Conclusion In 2015, Chinese adult residents had higher dietary sodium intake with gender and regional differences. Condiments were the main source of dietary sodium.