With the development of functional genomics, high throughput analysis of genes’ function has been the mainstream of research, and exogenous gene's over expression via Agrobacterium-mediated transformation is the most commonly used method in gene functional analysis.The versatile plant expression vector cassette named pBHT-5 was constructed by the method of site-specific mutagenesis based on pBI121. First of all, the restriction enzyme SfiI recognition site in trfA gene (X00713) which was relevant to plasmid replication and stability was replaced without changing its amino acid composition. And then the SfiIA,SfiIB sites were added between promoter CaMV35s and terminator NOS. The versatile plant expression vector cassette can be directly used to construct plant expression vector containing the full-length genes cloned by Clontech SMARTTM technology, which will raise the efficiency of vector construction. The result will provide basis of new genes’ high throughput screening and functional analysis, then get the new genes functioning in plant osmotic stress resistance.

Objective To find an ideal liver cirrhosis portal hypertension model in big animals suitable for surgical study.Methods Twenty canines were treated by subcutaneous injection of 60% CCl4 colza oil emulsion into the back,at a dose of 1.0-1.3 mL/kg every 10 days for a total of 6-8 times.At the same time,it was combined with the control of food intake.All canines were fed with rice mixed with 10% hog fat.The amount of rice was 15 g/kg per day from the first day to the forth day,but the amount of food was not controlled from the fifth day to the tenth day.During the process,the canines′ general condition was observed and the changes of hepatic functions such as ALT,TP,ALB,G and A/G were measured.The changes of liver morphology,the diameter and blood flow of portal vein were monitored with color Doppler.Every two weeks,a piece of hepatic tissue to observe the pathological change of liver was excised operatively.Results During the process,the body weight of the canines decreased continually;ALT increased during early and middle stages and decreased during advanced stage;TP and ALB always decreased,and A/G continuously decreased,but G did not change significamtly.Eight weeks after injection,liver became progressively smaller.its density was not well-distributed,and liver particles gradually became coarse,and liver capsule became rough.The diameter of portal veins became larger and the velocity of portal vein became slower.Liver cirrhosis with psudolobules was found on light microscopy from the tenth week to the twelfth week.Three canines died,with mortality of 15.0%.Conclusions It took 10-12 weeks to establish this liver cirrhosis portal hypertension models in canines by subcutaneous injection of CCl4.This method is convenient,has high success rate,and is suitable for use in surgieal research.

Objectives To investigate the expression of B lymphocyte stimulator (BLyS) and its receptor BAFF-R(a receptor of B-cell activator factor, belonging to the TNF family) in peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE), and to explore their clinical significance. Methods The patients were separated into active group (n = 28) and inactive group (n = 24) according to the SLE disease activity index (SLEDAI). The mRNA and protein expression of BLyS and its receptor BAFF-R were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western-blot in PBMCs from the patients and 21 healthy volunteers. The relationship between the expression of BLyS and BAFF-R and SLEDAI was analyzed. Results In the case of the expression of mRNA and protein of BLyS and BAFF-R, the patients had a higher level than the healthy controls (P 0.05). Conclusions These findings indicate that BLyS and its receptor BAFF-R might be involved in the pathogenesis of SLE. Also, BLyS expression level might be a new parameter for the evaluation of SLE disease activity and therapeutic effect.

Objective To investigate the relationship between cytomegalovirus infection and biliary atresia.Methods Sixteen infants with CMV infection and biliary atresia were retrospectively reviewed and compared with clinical manifestations with 29 infants with CMV hepatitis.Results Both serum CMV-IgM antibody and CMV-pp65 in polymophnuclear leukocytes were found positive in 9 cases,only CMV IgM antibody was found positive in 1 case and only CMV-pp65 were found positive in 3 cases,both CMV-IgM and CMV-pp65 were found negative in 3 cases.14 of 15 cases of CMV-pp65 were detected in liver tissues including 3 cases with negative CMV-IgM antibody and CMV-pp65 in blood.The clinical indexes in these 16 cases were much more severe than which in 29 infants with CMV hepatitis (P

Objective To investigate the correlation between age and the prognosis of thrombolytic therapy in acute ischemic stroke (AIS).Methods One hundred and fourteen patients with AIS were divided into ≤60 years group,61-70 years group and ≥71 years group according to age.Thrombolysis and post-thrombolysis treatment was done in accordance with 2010 version of Chinese Acute Ischemic Stroke Treatment Guidelines standard.The United States National Institutes of Health Stroke Scale (NIHSS) score was done in patient immediately after treatment,24 h after thrombolysis and 7 d after thrombolysis,and modified Rankin scale (mRS) score was assessed 3 months after thrombolysis The spontaneous intracranial hemorrhage (sICH) and the death of 2 weeks was recorded.Results ≤ 60 years group had 22 males and 10 females;61-70 years group had 26 males and 10 females; ≥71 years group had 20 males and 26 females.In ≥ 71 years group,women accounted for 56.52％ (26/46),which was higher than that in the other 2 groups,and there was significant difference (x2 =0.685,P =0.015).The NIHSS score immediately after treatment,24 h after thrombolysis and 7 d after thrombolysis among 3 groups had no significant difference (P ＞ 0.05).The mRS score at the 3 months after thrombolysis among 3 groups was (1 ± 3),(2 ± 5) and (2 ± 3) scores,respectively,and there was significant difference(P =0.040).Mortality and incidence of sICH in 2 weeks also had significant difference (P =0.049,0.017).Conclusions Despite the differences in the mortality and incidence of sICE among different ages,thrombolytic therapy with recombinant tissue-type plasminogen activator can significantly improve the neurological deficit after 3 months in AIS patients of different ages.

Objective To evaluate the iodine nutritional condition among populations in Aksu after implementing free iodized salt in population of Xinjiang.Methods According to the National Iodine Deficiency Monitoring Programme (Revised),totally 45 villages (towns) of subordinated 8 counties and Aksu City,Xinjiang were selected for collecting salt samples.Meanwhile,in 27 villages (towns) primary schools,school-aged children (8-10 years old) were selected for collecting disposable urine samples,B ultrasound was used to check thyroid volume and intelligence (IQ) was evaluated by China Combined Raven Test (CRT).Pregnant women were sampled for collecting their disposable urine near the primary schools.Both the children and pregnant women were evaluated the iodine deficiency disorders (IDD) knowledge awareness by questionnaire.The urine iodine concentration was measured by the ammonium persulfate digestion-As-Ce catalytic spectrophotometer method.The salt iodine concentration was measured by the direct titrimetric method,and other kinds of iodine in salt were measured by referee method (GB/T 13025.7-2012).Results A total of 2 700 household salt samples were collected,the average of iodine concentration in household salt was (26.95 ± 5.10) mg/kg.The coverage of household iodized salt was 98.56％ (2 661/2 700).The coverage of edible iodized salt was 98.00％ (2 646/2 700).A total of 2 159 urine samples were collected,the median of urine iodine concentration (UIC) among school-aged children (8-10 years old) was 235.50 μg/L.The total goiter rate was 1.51％ (33/2 179) among children aged 8-10 years old.A total of 2 098 people were conducted IQ test,the average IQ was 88.03 ± 17.14.A total of 1 047 urine samples were collected,the median of UIC among pregnant women was 213.50 μg/L.The IDD knowledge awareness rate of children and women were 98.82％ (751/760) and 99.23％ (258/260),respectively.Conclusions The iodine nutrition is in an adequate range,awareness of IDD prevention is obviously improved among children which is representative of common people and pregnant women after implementing the free iodized salt in population of Xinjiang in Aksu.The policy of preventing IDD based on this policy has showed enormous effect.

Objective To understand whether the breeding time of snails influences the evaluation on molluscicidal efficacy against Oncomelania snails so as to make a standardization of methods for screening molluscicides in laboratory. Methods Snails collected from the endemic areas in Nanjing and Tongling cities bred for 1,6,11,16,21.31,61 and 91 days respectively, and then were immersed in 1.000 0,0.500 0,0.250 0,0.125 0,0.062 5 and 0.031 3 mg/L solution of niclosamide for 24 and 48 hours at 25℃ in laboratory. Results One hundred percent killing rate was achieved in the groups of the snails bred for 1 - 11 days and immersed in 1.0 mg/L niclosamide for 24 hours. The LC_(50) concentrations of snails collected from Nanjing, Jiangsu Province and immersed for 24 hours varied from 0.094 7 to 0.133 9 mg/L, and for 48 hours from 0.071 8 to 0.092 6 mg/L.Those of snails collected from Tongling, Anhui Province for 24 hours varied from 0.082 5 to 0.103 9 mg/L, and for 48 hours from 0. 071 8 to 0.082 5 mg/L. The molluscicidal effect was unstable in the groups of snails bred for more than 16 days. There was a significant difference (X_(Nanjing24 h)~2 = 22.26,P

Objective To observe the impact of water temperature on shedding and infectivity of the cercariea of Schistosoma japonicum under laboratory condition. Methods Infected snails were exposed to 1, 5, 10, 15, 20, 25, 30, 35℃ and 40℃ water for 4 hours for shedding of cercariae respectively, the shedding rates and total numbers of cercariae shed were investigated. The mice were infected with cercariae in 1, 10, 20, 30℃ and 40℃ water for 30 minutes respectively, the infection rates and mean worm burden of mice were investigated. Results The cercariae were shed from infected snails with Jiangsu or Yunnan isolate of [WT5”BX]S. japonicum in water ranged from 1℃ to 40℃, but the fittest water temperatures range for shedding of cercariae of Yunnan isolate of S.japonicum[WT5”BZ] is from 20℃ to 35℃, while fittest range for Jiangsu isolate is from 15℃ to 35℃. All of the infection rates of mice infected with cercariae of Jiangsu isolate of S.japonicum in the water ranged from 1℃ to 40℃ were 100.0%. Infection rates of mice infected with cercariae of Yunnan isolate of S. japonicum in the water ranged from 1℃ to 40℃ were more than 83.3%. Conclusions When infected snails are exposed to the water with temperature ranged from 1℃ to 40℃ it is possible for cercariae to shed and infect the final hosts . It is suggested that there is a possibility of infection with S.japonicum when contacting the water near marshlands with infected snails in winter.

Objective To enhance the protective immunity effects against Schistosoma japonicum infection by priming with cocktail DNA vaccines and boosting with cocktail protein vaccines in infected BALB/c mice.Methods Plasmids and proteins for immunization were prepared and diluted in no bacterial saline solution to final concentration of 1.5 mg/ml,and mixed with pcDNA3.1-SjC23,pcDNA3.1-SjCTPI,pcDNA3.1-(CDR3)6 plasmid DNAs by equal volume to form the cocktail DNA vaccine,and also mixed with recombinant proteins SjC23-HD,SjCTPI,and NP30 by equal volume to form the cocktail protein vaccine.Seventy female BALB/c mice of 4-5 weeks old were randomly divided into 5 groups(A,B,C,D,E).In Group A(control group),each mouse was immunized with 100 ?l saline solution by intramuscular(i.m.);in Group B(pcDNA3.1 control group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6;in Group C(pcDNA3.1 and cocktail protein group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9;in Group D(cocktail DNA vaccines group),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6;in Group E(cocktail DNA vaccines plus cocktail proteins),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9.Four weeks after the last DNA immunization or two weeks after protein boosting,all the mice were challenged with(40?1)cercariae of Schistosoma japonicum by abdominal skin penetration at the same time.Forty-two days post-challenge,the mice were sacrificed and perfused,and the numbers of recovered worms and eggs in liver were counted.The blood was collected from the tail veins of all the mice two days before the first immunization and challenge,respectively,the serum was prepared for detection of IgG,IgG1 and IgG2a.Two days before the challenge,the spleen cells of two mice from each group were cultured and stimulated with ConA and soluble egg antigen(SEA),and the supernatant was collected for detection of IL-2,IL-4 and IFN-?.Results The worm reduction rates in Group C,D and E were 17.70%,32.88% and 45.35%,respectively,compared with the control group.The worm reduction rates in Group D and E were significantly higher than that in Group C(P

OBJECTIVETo investigate the dynamic expression and role of SENP1 (SUMO-specific proteases-1) in the pulmonary vascular wall of rat during the development of hypoxic pulmonary hypertension (HPH).

METHODSForty adult male Wistar rats were randomly divided into 5 groups (n = 8), and exposed to normoxia (Control group) or exposed to hypoxia for 3, 7, 14 or 21 d, respectively. The HPH models were established by normobaric intermittent hypoxia. Mean pulmonary arterial pressure (mPAP), right ventricle hypertrophy index (RVHI), and vessel morphometry were measured. Reverse transcriptase-polymerase chain reaction(RT-PCR) and in situ hybridization were used to determine the mRNA expression of SENP1. Immunohistochemistry and Western blot were used to determine the protein expression of SENP1.

RESULTSThe hypoxic rats developed pulmonary vascular remodeling in pulmonary arterioles after 7 d of hypoxia exposure. Pulmonary vascular remodeling in pulmonary arterioles significantly increased after 14 d of hypoxia. The level of mPAP in hypoxic rats increased significantly after 7 d of hypoxia, reached its peak after 14 d of hypoxic exposure. RVHI was markedly increased after 14 d of hypoxia. In situ hybridization and immunohistochemical analysis showed that SENP1 mRNA and protein were positively stained in control. SENP1 mRNA expression had little changes after exposure to hypoxia compared with the control, however, SENP1 protein expression was declined gradually after 7 d of hypoxia. The results of RT-PCR and Western blot showed that the same dynamic expression of SENP1 mRNA and protein in lung tissues of rats. Linear correlation analysis showed that SENP1 protein were negatively correlated with mPAP, pulmonary vascular remodeling index and RVHI.

CONCLUSIONUnder chronic hypoxia, SENP1 protein can be degradated. The dynamic expression of SENP1 protein may play a role in implicating in the development of HPH.

Animals ; Endopeptidases ; metabolism ; Hypertension, Pulmonary ; etiology ; metabolism ; Hypoxia ; complications ; metabolism ; Male ; Pulmonary Artery ; metabolism ; Rats ; Rats, Wistar