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.

sing guidewire through temporary catheter is an alternative strategy, because of avoiding this avoids vein re-puncturation and doesn't increase catheter related complication.

Objective To evaluate the efficacy of endoscopic ultrasonography(EUS)guided implantation of 5-FU seeds for progressive gastric cancer.Methods A total of 23 patients with progressive gastric cancer and outflow obstruction were enrolled into the study.During the endoscopy procedure, 45-60 5-FU seeds were implanted into the tumors with guidance of EUS, with an average dose of 100 mg.Volumes of tumor were examined with 3D-EUS before and after the treatment.Results A total of 33 procedures of 5-FU implantation were performed.By measurements of 3D-EUS, the numbers of patients with complete respons, partial response, no change, progressive disease were 1, 15, 6 and 1, respectively.In terms of outflow obstruction improvement, significant effect, obvious effect and average effect were achieved in 6, 7 and 9 patients, respectively, with an overall response rate at 91.4%.Conclusion EUS-guided 5-FU seed implantation in progressive gastric cancer is safe,reliable and effective with few complications.3D-EUS may be a new method to evaluate curative effect.

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.

Objective To compare the effect of percutaneons coronary interventions (PCI) combined with polydanshinolate on myocardial reperfusion with PCI combined with regular medicine in patients with acute myocardi-al infrarction ,and whether polydanshinolate would decrease no-reflow,improve the myocardial micro-circulation and the ventricular remodeling and clinical end-point events at the sixth month. Methods Random,parallel control and prospective clinical design was used in the current study. Sixty eligible patients for PCI, who were diagnosed as first-time acute myocardial infrarction with ST stage increasing,were recruited in the study with informed consent. All par-ticipants were divided into two groups randomly. Group A was treated with PCI combined with polydanshinolate, group B was treated with PCI combined with regular medicine. The participants were followed up for six months. The resolution of the sum of ST segment elevation (sum STR) ,corrected TIMI frame count (CTFC) and myocardial con-trast echocardiography (MCE) were used to assess myocardial perfusion. Bleeding events, heart function and major adverse cardiac events (MACE) were observed during hospitalization and follow-up visit. Results No significant difference was observed between two groups on the clinical condition and the results of emergency CAG and PCI. Compared to group B,the incidence of TIMI grade 3 was significantly higher in group A (90.0% vs 63.3% ,χ~2 = 4.565, P=0.0326). Sum STR one hour after PCI in group A was siguificanfly higher than that in group B (80.0% vs 50.0%, χ~2=4.689, P=0.0304). CTFC after PCI also differed significantly between the two groups (24.1±8.3 vs 33.4±15.9 respectively,P=0.0062). Localized myocardial blood flow 48 hours after PCI showed no significant difference between the two groups (P>0.05), whereas both increased at the 7th day after PCI (5.85±1.26 vs 2.09±1.85,t=9.2008,P<0.0001 ;3.95±1.35 vs 1.95±1.29,t=5.8666,P<0.0001) ,and there were signifi-cant difference between the two groups (P<0.05). No significant difference of LVEF, LVEDV, LVESV were found between the two groups during the hospitalization and follow-up visit after 6 months (P>0.05). The incidences of cardiovascular events and severe heart failure during hospitalization and follow-up visit in group A were significantly lower than that in group B (P<0.05 ). Conclusions Polydanshinolate addition to PCI could reduce no reflow phe-nomenan in patients of acute myocardial infrarction, improve myocardial reperfusion, and furthermore, decrease cardi-ovascular events and ameliorate prognosis.

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 .

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.