The aim was to establish an effective screening microarray at genus level for Pospiviroid. We analyzed nucleotide sequences from Pospiviroid viroid and designed 19 probes with genus identification characteristics. The standards of these probes included the characters of (i) a GC content between 40 and 60%, (ii) less than 50% of single nucleotide, (iii) less than 4 continuous mononucleotides, and (iv) less than 6 nucleotides in the inner hairpin. We synthesized microarrays by using these probes on glass slides. The validation results of microarray probes show effective signals from chrysanthemum stunt viroid and tomato planta macho viroid standard samples hybridization. The sensitivity results show that the microarray detected 200 pg/microL of total RNA. The microarray can be used to screen Pospiviroid viroid.
Base Composition ; Base Sequence ; Microarray Analysis ; Nucleic Acid Hybridization ; Plant Diseases ; virology ; Plant Viruses ; classification ; RNA ; Viroids ; classification

Objective To study the imaging features of sacroiliac joints(SIJ) in patients who were misdiagnosed as spondyloarthritis(SpA).Methods A total of 34 patients with chief complaint of back pain and misdiagnosed as SpA from January 2007 to April 2013 in Department of Rheumatology Chinese PLA General Hospital were enrolled.The imaging,clinical manifestations,laboratory examinations data were analyzed.Result The main reason for misdiagnosis as SpA was because of sacroiliitis presenting on imaging.The final diagnoses included 24 patients as SIJ infection,4 patients as neoplastic diseases,2 patients as metabolic bone diseases,2 patients as sacroiliac joint degeneration,1 patient as gout of sacroiliac joint,1 patient as diffuse idiopathic bone hypertrophy.For patients with infection,there were 10 patients receiving X-ray and 22 patients receiving CT of SIJ.However,5 and 7 patients had negative results respectively.These patients with infection had abnormalities in MRI including all with bone marrow edema,21 patients with erosion of bone and joint,22 patients with muscle involved.As to the patients with malignancies,SIJ CT scan appeared normal.Bone marrow edema and erosion in MRI were found in all neoplasm patients expect one as ependymoma.Adjacent muscles were involved in the patient with Ewing's sarcoma.Either X-ray or CT in other patients demonstrated obvious abnormalities,but only mild erosion of bone was found in MRI.Conclusion Bone marrow edema of SIJ in MRI represented not only in patients with SpA.Rheumatologists should analyze the clinical manifestations and laboratory examinations comprehensively in order to avoid the misdiagnoses.

Objective To construct the recombinant protein AtlM ( rAtlM) of Staphylococcal au-reus through prokaryotic expression system and to investigate its antibacterial activity in vitro.Methods The specific primers were designed according to atlM gene sequence of Staphylococcal aureus recorded in Gen-Bank, and atlM gene was amplified by PCR from the Staphylococcal aureus strain (ATCC25923).The re-combinant plasmid pET-32а(+)/atlM was constructed and transformed into Transetta ( DE3 ) to express AtlM after induced by IPTG .The expressed protein AtlM was further analyzed by SDS-PAGE and purified by electroeluting of bag filter.The minimal inhibitory concentrations (MICs) of rAtlM to ATCC25923 and oxac-illin-resistant S.aureus strain were determined by the broth microdilution method .S.aureus ATCC25923 strain and oxacillin-resistant S.aureus strain (final concentration of 5×105 CFU/ml) were exposed to rAtlM (50 μg/ml) respectively to test its antibacterial activity in vitro.Results The recombinant protein AtlM was expressed and purified successfully with a relative molecular weight of 80 ×103 and a concentration of 1.25 mg/L.The MICs of rAtlM to ATCC25923 strain and oxacillin-resistant S.aureus strain were 8 μg/ml and 64 μg/ml, respectively.In vitro test showed that rAtlM had inhibitory effects on the growth of ATCC25923 strain and oxacillin-resistant S.aureus strain after 1 h of intervention (P=0.004 and P=0.026, respectively), which lasted to 5 h for ATCC25923 strain (P=0.012) and 3 h for oxacillin-resistant strain (P=0.001).Conclusion This study shows that rAtlM has a certain antibacterial effects on S.aureus ATCC25923 strain and oxacillin-resistant S.aureus strain, suggesting a possibility of serving as antimicrobial agent.

Objective To obtain the pneumococcal autolysin(LytA)and choline binding protein A(CbpA)by prokaryotic expression system and investigate their diagnosis for infection caused by Streptococcus pneumoniae.Methods The specific primers were designed according to lytA and cbpA of Streptococcus pneumoniae gene sequence.lytA and cbpA were amplified by PCR form the pneumococcus genome.After IPTG inducing,the recombinant proteins were purified by electroeluting of bag filter,detected by SDS-PAGE and Western blot.Serum lgG and IgM antibodies accordingly of BALB/c mice infected with Streptococcus pneurnoniae were detected by ELISA.Results The recombinant plasmid pET-32a(+)/lytA and pET-32a (+)/cbpA were constructed successfully.Fusion proteins LytA and CbpA were expressed and displayed expected antigenicity.IgM and IgG antibodies level anti LytA were significantly higher than the control group (infections with B Streptococcus group and healthy mice),(P<0.05),but antibodies level anti CbpA did not increase as compared with group infected with B Streptococcus(P>0.05).Diagnostic sensitivity of CbpA was 83.3%(IgG)and 75.0%(IgM).Diagnostic specificity of LytA was 100%(IgG and IgM).Conclusion The synergistic use of specificity of LytA and sensitivity of CbpA may be worthy of serological diagnosis for Streptococcus pneumoniae infection,and may be used for further clinical test.

Objective To investigate the effects of ulinastatin on renal expression of endothelin￣1 and nitric oxide (NO) in rats with toxic acute kidney injury(AKI). Methods Twenty￣four male SD rats were randomly divided into the normal control group,model control group and treatment groups, with 8 rats in each group. Except normal control group, rats were subcutaneously injected with gentamicin (300 mg.kg-1 .d-1 ) for 3 days to establish a model of toxic AKI.Rats in the treatment group were intraperitoneally injected with a 7￣day course of ulinastatin (30 000 U.kg-1 .d-1 ) from the fourth day.While the other two groups were injected with 0.9% sodium chloride injection (3 mL.kg-1 .d-1 ). The serum level of creatinine and cystatin￣C,urinary concentration of kidney injury molecule￣1(Kim￣1) and neutrophil gelatinase￣associated lipocalin (NGAL), level of endothelin￣1 and NO,expression of endothelin￣1 mRNA,endothelial nitric oxide synthase (eNOS),induced nitric oxide synthase (iNOS),eNOS mRNA and iNOS mRNA in homogenate of renal tissues in each group were detected on the eleventh day. Results Compared with the normal control group,serum level of creatinine and Cystatin￣C,urinary concentration of Kim￣1 and NGAL,level of endothelin￣1 and NO,expression of endothelin￣1 mRNA,iNOS and iNOS mRNA in homogenate of renal tissues were higher in model control group (P<0.01, respectively), while which were lower in treatment group than those in model control group ( P < 0. 01, respectively). And no statistical significant difference of eNOS and eNOS mRNA expression in homogenate of renal tissues existed among the three groups. Conclusion Ulinastatin possesses curative role against rat with toxic AKI via down￣regulating renal expression of endothelin￣1,NO and iNOS.

Objective To study the relationship between gastritis cystica profunda (GCP)and early gastric cancer (EGC),and to explore the diagnosis and treatment of GCP.Methods Clinical data of 17 patients,who were pathologically diagnosed as having GCP,including clinical symptoms,white light endo-scopic findings,magnifying endoscopic features,EUS outcomes and histopathological characteristics were an-alyzed.Results All 17 patients of GCP had no specific clinical symptoms or history of gastric surgery prior to the operation.Eight lesions were located at cardiac,4 at gastric body,4 at gastric antrum and 1 at gastric angle.As to the morphology,3 cases were type Ⅰ,3 type Ⅰ +Ⅱc,9 type Ⅱa +Ⅱc,1 type Ⅱc and 1 type Ⅱb +Ⅱc.The mean lesion size was 1.8 cm.Of 16 patients with preoperative EUS examination,11 patients were found to have cystic changes,with the diagnostic rate being 68.8%(11 /16).All 17 patients underwent ESD,14 (82.4%,14 /17)of which were found to have EGC.Postoperative pathology confirmed low grade intraepithelial neoplasia in 3 cases,high grade intraepithelial neoplasia in 7,intramucosal carcino-ma in 4,submucosal carcinoma in 3 patients.Conclusion EUS plays an important role in the diagnosis and treatment of GCP.And GCP is highly correlated with EGC,which should be managed in time.

ObejectiveTo evaluate the efficacyofacupointthread embedding in easing painafterMilligan-Morgan(M-M)for mixed hemorrhoids.MethodSixty patients undergone M-M for mixed hemorrhoids were randomized into a treatment group of 30 cases and a control group of 30 cases. After M-M, patients in the treatment group received thread embedding at Changqiang (GV1) and bilateral Zhibian (BL54), while the control groupdidn’treceive any intervention. The onset time of post-operative pain, average pain index within a week, and pain index after defecation, electromyogram (EMG), change of anal canal pressure, patients’ satisfaction, and adverse-event rate were observed.ResultThe average pain index and pain index after defecation in the treatment group were significantly lower than that in the control group (P<0.05), and the onset of pain in the treatment group was significantly later than that in the control group (P<0.01); after surgery, the anal canal resting pressure in the treatment group was markedly lower than that in the control group (P<0.05); there wasno significant difference in comparing the squeeze pressure of anal canal between the two groups (P>0.05). According to the motor unit potential (MUP) analysis, there were significant differences in comparing the amplitude (Ampl) and Ar/Am of the restingphase between the two groups (P<0.05), while there were no significant differences in comparingthe Ampl, Area, Ar/Am, and Freq of the contraction phase between the two groups (P>0.05). There were significant differences in comparing the patients’satisfaction, adverse-event rate, and use of analgesics between the two groups (P<0.05). ConclusionAcupoint thread embedding can produce a content analgesic effect, and it’s safe and reliable.

Objective To investigate the incidence of sub-health fatigue among medical workers exposing to radioactive rays, and to observe the therapeutic effect of Bushen Jianpi recipe on sub-health fatigue. Methods An investigation was carried out among 1 056 medical workers exposing to radioactive rays which were from the medical and sanitary institutions of Guangzhou city. Sixty cases subjecting to the inclusion criteria were sampled from the investigated population by epidemiological methods for the randomized controlled trial. The subjects were evenly randomized into treatment group and control group. Both groups received health education and health-promoting lifestyle, and the treatment group was given self-prescribed Bushen Jianpi recipe additionally. The treatment lasted for 4 weeks. The scores of Multidimensional Fatigue Inventory ( MFI-20) and Sub-health Measurement Scale Version 1.0 ( SHMS V1.0) were observed before and after treatment, and the therapeutic effect was evaluated after treatment. Results ( 1) We got 976 effective questionnaires, and the effective response rate was 92.4%. Among the admitted 976 workers, the incidence of sub-health state was 66.8% (652/976) and that of sub-health fatigue is 31.4% ( 205/976). ( 2) The total effective rate was 80.0% in the treatment group and was 36.7% in the control group, the difference being significant ( P<0.01). ( 3) After treatment, the scores of MFI-20 and SHMS V1.0 were decreased in the treatmeat group, and fatigue symptom scores were decreased in the control group ( P<0.05 or P<0.01) , the decrease in the treatment group being superior to that in the control group ( P<0.05 or P<0.01). Conclusion The incidence of sub-health fatigue is popular in the medical workers exposing to radioactive rays, and Bushen Jianpi recipe has an effect on improving the sub-health fatigue.

Objective To explore the curative effect of ulinastatin against toxic acute kidney injury(AKI) in rats and its mecha-nism .Methods Twenty-four male SD(Sprague Dawley) rats were randomly divided into 3 groups ,control group ,model group and treatment group with 8 rats in each group .Rats were subcutaneously injected gentamicin(300 mg/kg of body weight per day) for 3 days to establish models of toxic AKI .Rats in treatment group were intraperitoneally injected with a 7-day course of ulinastatin(30 000 U/kg of body weight per day) from 4th day .Dectetion of serum level of creatinine and Cystatin-C(Cys C) ,urinary concentra-tion of kidney injury molecule-1 (Kim-1 ) and neutrophil gelatinase-associated lipocalin (NGAL ) ,activity of superoxide dismutase (SOD) and glutathione peroxidase(GSH-Px) ,content of malondialdehyde ,levels of tumour necrosis factor-alpha(TNF-α) and inter-leukin-1β(IL-1β) in homogenate of renal tissues as well as observation of renal pathological changes and semiquantitative score in each group were conducted on 11th day .Results In model group ,degeneration and necrosis of renal tubular epithelial cell ,dilatation of renal tubular cavity and inflammatory cell infiltration in renal interstitial were observed .Renal pathological changes were milder in treatment group ,when compared with the model group .Renal pathological semiquantitative score ,serum level of creatinine and Cys C ,urinary concentration of Kim-1 and NGAL ,content of malondialdehyde ,levels of TNF-α and IL-1β in homogenate of renal tissues were higher in model group than in control group ,while those in treatment group were lower than in model group(P<0 .01 , respectively) .And activity of SOD and GSH-Px in homogenate of renal tissues were lower in model group than in control group ,and those in treatment group were higher than in model group and control group(P<0 .01 ,respectively) .Conclusion Ulinastatin pos-sesses a curative role in toxic AKI in rat via inhibiting oxidative stress and down-regulating levels of proinflammatory factor in renal tissues .

Objective To explore the value of combined measurement of urine N-acetyl-beta-D-glucosaminidase (NAG) activity and serum Cystatin C in diagnosing diabetic nephropathy (DN) in early phase. Methods Sixty-two cases with type 2 diabetes (diabetic group) were divided into three groups according to their 24 hours urinary albumin excretion (24hUAE) : group A (normal albuminuria, 20 cases), group B (microalbuminuria, 22 cases) and group C (macroalbuminuria, 20 cases). Furthermore, 30 healthy people were involved in control group. 24hUAE,NAG,serum creatinine (SCr) and serum Cystatin C were measured, and endogenous creatinine clearance rate (Ccr) was calculated by Cockcroft-Gault formula. All these indexes among three groups were compared. Results The levels of urinary NAG activity and serum Cystafin C in diabetic group was significantly higher and Ccr was significantly lower than those in control group(P < 0.01). The levels of urinary NAG activity and serum cystatin C gradually increased in group A, B and C(P< 0.05 or < 0.01). While no significant difference was observed between group A and group B in the level of SCr (P > 0.05). There were significant positive correlations among the levels of urinary NAG activity, serum Cystatin C,24hUAE and SCr (P< 0.01),and all above showed negative correlations with Ccr (P<0.01). Co-detection of urinary NAG activity and serum Cystatin C had significantly higher positive rate [80.6%(50/62)] than single one [58.1%(36/62),61.3%(38/62)](P<0.05). Conclusion Co-detection of urinary NAG activity and serum Cystatin C may indicate early renal damage in DN, and it is valuable in diagnosing DN in early phase.