NS2 and NS3 are two post-transcriptional gene silencing suppressors that are encoded by Rice stripe virus. Gene silencing suppressors are always related to the pathogenicity of viruses. In this study, the cDNA of NS2 and NS3 were recombined by overlapping PCR assays, ligated to the RNAi vector, and inserted into the PXQ expression vector using Pst I; the expressed vector was transferred into calluses induced from seeds of the japonica rice cultivar, 'Nipponbare', using an Agrobacterium-mediated method. Thirty-one T0 transgenic plants were selected by G418 screening. PCR and southern blot analyses confirmed that the target gene was transformed into transgenic rice successfully, and different transgenic plants contained various copies of the gene. The disease resistance assay revealed that T0 transgenic rice had a delayed onset of RSV for approximately 10-20 d, and the accumulation of virus in the transgenic plants was reduced by 30%-50%. This was related to the delayed onset of disease.
Disease Resistance ; Oryza ; genetics ; immunology ; virology ; Plant Diseases ; genetics ; immunology ; virology ; Plants, Genetically Modified ; genetics ; immunology ; virology ; RNA Interference ; Tenuivirus ; genetics ; immunology ; Viral Nonstructural Proteins ; genetics ; immunology

Objective To investigate a special optimization technique for computer aided measure, and provide anatomical basis for screw internal fixation in the cavitas glenoidalis through the coracoid process of scapula. Methods Thirty accurate scapula models were reconstructed from CT data sets. First, special optimization objective function was designed for single screw internal fixation configuration, and the optimal placement of screw was found automatically under constraints. Then, the placements of double screws internal fixation configuration were searched taking advantage of principal component analysis. Finally, statistical measure data were provided according to new anatomical reference landmarks for clinical use. Results For single screw internal fixation configuration, the distance from the optimal screw entry point P to the acromion process point X was (39.15±2.28) mm, to the coracoid process point Y was (28.66±2.68) mm, to the angulus superior point Z was (61.13±6.57) mm;The angle was (81.27±7.15)° between PX and PY, and (133.27±6.84)° between PX and PZ. The mean inclination of the lag screw was (104.08±4.41)° for the angle with line PX, (101.29±3.51)° with line PY, and (76.23±5.03)° with line PZ. For double screws configuration, the distance from the original single screw entry point P to the screw entry point E was (5.12±1.37)mm,to the screw entry point F was (3.88±0.94)mm. The angle between the long axis of coracoid process and line EF was (27.41±3.51)°. Conclusion The automatic optimization technique for computer aided measure is very efficient and has many advantages over the conventional manual dissection methods, and is convenient to design new anatomical reference landmark system for clinical use.

Highly Active Anti-Retroviral Therapy (HAART) has effectively inhibited the prevalence of HIV-1 and reduced the death rate caused by AIDS. In recent years,the emergence of resistance-conferring RT gene mutations in HIV-1 strains has become the major reason for HAART failure. The detection of drug resistance is important for the HAART regimen choice and novel drug development. A novel assay for the detection of HIV-1 RT drug resistance mutations was developed. HIV-1 drug resistance and wild strains in B subtypes were investigated using Two-Step Mutagenically-Separated PCR (MS-PCR),and point mutations including M41L,K70R,K103N,Y181C,T215F were detected. A longer mutant type primer was designed,using microplates hybridization and ELISA technique to detect several point mutations within a mixed mutant-wild type population. The results indicate that the Two-Step MS-PCR is as sensitive and specific as that in the traditional MS-PCR and MS-PCR combined with ELISA can give a good P/N quotient with better sensitivity,low cost,relatively less time consumption and high-throughput screening. It will be used in clinic usage for the detection of HIV-1 drug resistance mutations as well as other point mutations.

HIV-1 envelope glycoprotein gp41,which is a hopeful target for HIV-1 fusion inhibitors,plays a critical role in the fusion of viral and cellular membranes.In order to build up the screening assay of HIV-1 fusion inhibitors targeting gp41,HIV-1 gp41 5-helix and 6-helix were expressed in prokaryotic cells.Gp41 5-helix and 6-helix recombined plasmids were constructed by using PCR,enzyme digestion and ligation taking the clade B HIV-1 genome as a template.The plasmid was transferred into E.coli BL21(DE3)and then induced by IPTG.The expressed protein was purified by affinity chromatography after denaturation and renaturation.The SDS-PAGE analysis was used during expression and purification.Native-PAGE was used to identify the interaction between gp41 5-helix and T-20.The result will be helpful to build up the screening assay of HIV-1 fusion inhibitors targeting gp41.

The pol gene of HIV-1 encodes mainly three enzymes: reverse transcriptase (RT), protease (PR) and integrase (IN). Currently, FDA approved drugs targeting RT and PR are available and administered in various combinations, while no anti-IN drug was approved. HIV-1 integrase is an essential enzyme for the viral replication and an interesting target for the design of new pharmaceuticals for multi-drug therapy of AIDS. The 288 amino acids of IN (32kDa) recognizes specific sequences in the long terminal repeats (LTRs) of the retroviral DNA. The IN protein catalyzes the 3′-processing step and the 5′-strand transfer step reaction in vivo, which was called integration and this reaction could be analysed by ELISA Assay in vitro. It has been reported that F185K and C280S mutations of HIV-1 integrase would improve the enzyme solubility, and the catalytical activity of the enzyme was the same as that of the wild-type enzyme in vitro. In order to build the platform of screening inhibitor against integrase of HIV-1 virus, the IN enzyme was expressed and the function of integrase protein was assayed. The cDNA of clade B HIV-1 genome was used as a template, overlapped PCR was used to construct site mutagenesis of F185K/C280S and NdeI/Xho I restriction sites were brought in. The PCR product was cloned into the prokaryotic vector pET-28a(+) to form a recombined plasmid, transferred into the host cell E.coli(BL21 DE3). The recombined clones were identified by PCR and Nde I/Xho I digestion .The positive plasmid was sequenced, and the successfully recombined plasmid in the host cell was induced by IPTG. The expressed IN protein was puriied sy the Co+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombined IN protein. The recombinant protein was soluble, and expressed highly and stably in E.coli. The molecular weight of the expression product was identical to the expectation. The IN protein was proved to be functional in 3′ processing and 5′strand transfer by ELISA. It will be helpful to build the platform of screening inhibitors against HIV-1 integrase.

Objective To study the effects of different level of iodine nutrition on the thyroid function in women of reproductive age. Methods A total of 100 (50 from each) women of reproductive age but not pregnant were collected from iodine deficient and adequate areas. The questionnaire was obtained individually with items concerning personal history of thyroid diseases, goiters and category of edible salt and drinking water based on the project design. The household salt and drinking water were collected for measuring iodine content, and blood samples were obtained for TSH, FT4 and FT3 testing. Results The coverage of iodized salt and the median level of urinary iodine in iodine deficient women(72.0% and 95.5 μg/L) were obviously lower than that in iodine sufficient women(100.0% and 167.4 μg/L, χ2=16.28, U = 632.00, P < 0.01). Median level of serum TSH in iodine deficient women (2.56 m U/L) appeared in an increasing tendency compared to the iodine sufficient women (1.88 mU/L), but there was no significance (U=990.50, P > 0.05). Serum FT4 mean level in iodine deficient women [(14.7±2.0) pmol/L]was lower than that in iodine sufficient women[(17.0±3.8)pmoI/L, t=3.76, P<0.01]. There was no difference in serum FT3 between two group women[(5.1±1.4), (4.8±0.5)pmoI/L, t = 1.59, P > 0.05]; but FT3/FT4 ratio in iodine deficient women(0.33±0.04) was markedly higher than that in the iodine sufficient women(0.30±0.04, t=3.13, P<0.01). The percentage of thyroid dysfunction in iodine deficient women[20.0% (10/50)]was higher compared with the iodine sufficient women[8.0%(4/50)], but without significance(χ2=2.99, P>0.05). Conclusions Iodine deficiency is a primary cause leading to hypothyroid in women of reproductive age. Long term of iodized salt usage is an efficient way to correct iodine deficiency.

OBJECTIVETo understand prevalence of workplace violence in hospital and to analyse its relevant causes to lay a basis for maintaining normal working order in hospital.

METHODSA study was conducted to look into workplace violence situation in health care workers in two large hospitals of Guangzhou, Guangdong Province during October 2001 to October 2002. Workplace violence was defined as any events occurred in hospital staff, who suffered psychological or/and physical violence during the past 12 months.

RESULTSTotally, 678 of 1 043 hospital staff (65%) investigated had such experience during the past year, mainly psychological violence. Medical doctors were more vulnerable than nurses, with prevalence of 70.3% and 67.7% for medical doctors and nurses, respectively. Prevalence was the highest in those aged 30 - 39 years with 11 - 20 years of employment. Man staff were more vulnerable to physical violence than women, with prevalence of 11.7% and 5.3%, respectively. No significant difference in psychological or sexual violence between man and woman staff was found. Frequently, nurses and nurse aides were victims of sexual violence. Usually, troublemakers were patients relatives or patients themselves, accounting for 64.2% and 50.0% of the total events, respectively. Main causes for workplace violence in hospital included unreasonable requirement from patients or their relatives which was not met, or not-so-quick recovery as they desired.

CONCLUSIONSWorkplace violence occurred in hospital staff was prevalent in Guangzhou, which should be attached more importance. Comprehensive intervention measures should be adopted focusing on law reinforcement and education, to maintain normal working order in hospital.

Female ; Humans ; Male ; Personnel, Hospital ; Prevalence ; Violence ; prevention & control ; statistics & numerical data ; Workplace

OBJECTIVETo collect basic information on family burdens and long-term influence of children suffered from traumatic brain injury (TBI).

METHODSThrough prospective study, child behavioral problems, and injury-related family burden were assessed longitudinally in children with TBI over 6 months during the post injury period and children's pre-injury family function rated by parents soon after injury. Post injury child behavior and family outcomes were assessed at 6-month follow-up period.

RESULTSThe mean adaptation partnership growth affection and resolve scale (APGAR) score of 113 children before TBI was 7.96 and score after TBI was 6.94, which had significantly difference through t test. The mean APGAR score after 6 months was 7.60, which was significantly different from the hospital data. Among group with severe TBI, the family APGAR score in hospital was significantly smaller than that before injury occurred, and the family APGAR score in 6 months after being discharged from the hospital had no significant difference with the score when staying in the hospital. The three leading dimensions among family burden scale of diseases (FBS) scores after TBI were dimension of family economic burden, family daily life and family entertainment. 6 months later, the three leading dimensions had changed to be as dimension of mental health status, dimension of family relationship and dimension of family economic burden. Mean score of child behavior checklist (CBCL) assessed at 6-months follow up period among 113 children was among normal range.

CONCLUSIONSFamily function of children with TBI was affected by TBI. However, family function could be recovered along with child's convalescence except among children with severe TBI. Long-term pressure of TBI on family was revealed in mental health status and family relationship. In this study, there were no evidence of association between TBI and children's behavior problem.

Brain Injuries ; complications ; physiopathology ; Child ; Child Behavior Disorders ; etiology ; China ; Cost of Illness ; Family Health ; Hospitalization ; Humans ; Prospective Studies ; Severity of Illness Index

Objective To study the molecular mechanisms of ampicillin- resistant haemophilius influenzae (Hi)in Nanjing. Methods One hundred and fifty- eight strains of Hi isolated from children were collected to detect bata-lactamase. TEM and ROB bata- lacta-mase genes were detected by polymerase chain reaction (PCR) ,and cloned into T vector for sequencing. Results The rate of ampicillin resistance was 41. 77% in Hi isolated from children in Nanjing,40.51 % was found to be bata-lactamase production. Eighty-nine strain were TEM positives, 1 strain was ROB positive,63 strains bata - lactamase positive ampicillin- resistant Hi were identified. The resistance mechanism of ampicillin resistant Hi was production of bata - lactamase , mainly TEM - type enzyme. Two bata - lactamase negative ampicillin - resistant Hi were identified , predicts the other mechanisms of ampicillin - resistant Hi was occuered yet . One strain of non -TEM - type,and non - ROB - type bata - lactamase - producing Hi was identified. Conclusions Ampicillin - resisitant in Hi isolated from children in this region is challenging. TEM bata - lactamase is the principal mechanism of ampicillin - resistant of Hi.

The technique conditions of decolonization of fermentation broth were successively optimized using single factor assay and orthogonal layout method in Cordyceps jiangxiensis. The optimal condition of decolorization was investigated to be 3g/100mL active carbon, 5 min absorption time, pH5 of fermented broth and 25℃absorption temperature. Under the optimal condition, the maximum decolorization rate of fermented broth reached 89. 6% , simultaneously 10. 7% consuming rate of exopolysaccahride was minimum. Subsequently, the extract condition of exopolysaccharide of C. jiangxiensis was further optimized by orthogonal layout design. The maximum exopolysaccharide production was 0. 38 g/L under the optimal condition, i. e. firstly fermented filtrate decolorized and deproteined was concentrated to 1/7 of its total volume, secondly concentration broth was mixed with four times its volume of absolute ethanol and stirred vigorously, lastly precipitation of exopolysaccharide proceeded at 4℃for 16 hrs and the exopolysaccharide collected by centrifugal ion and dryness.