OBJECTIVE To establishan immuno-PCR assay with the carriers of gold-magnetic particles for detection of HIV-1 p24. METHODS The feasibility of using gold-magnetic particles as the carriers was verified. The gold-magnetic particles were coated with mouse anti-p24 monoclonal antibody as the capture antibody. The reporter DNA was initially generated by PCR amplification using a biotinylated primer, and was bound through streptavidin to biotinylated polyclonal antibody as the detection antibody. HIV-1 p24 sandwiched by two antibodies was detected by amplifying the reporter DNA using PCR. RESULTS The efficiency of gold-magnetic particles coated with mouse anti-p24 monoclonal antibody could reach up to 95%. Furthermore, the amount of antibodies immobilization was consistent among different batches of gold-magnetic particles and there was nearly without nonspecific adsorption. The detection limit of immuno-PCR assay was 0.1 ng/L, an approximately 1.5?104-fold higher compared with an enzyme-linked immunosorbent assay. The linear range of p24 concentration was 0.1-100 ng/L. CONCLUSIONS Gold-magnetic particle is one of the ideal immuno-PCR reaction carriers. The immuno-PCR for detection of HIV-1 p24 reported in this article is indicated to be a promising detection method.

Objective To describe a highly sensitive immuno-polymerase chain reaction (immuno-PCR) assay for the detection of human recombinant HIV-p24 antigen. Methods We used gold-magnetic particles as the carriers,mouse anti-p24 monoclonal antibody as the capture antibody and biotinylated goat anti-p24 polyclonal antibody as the detection antibody. The reporter DNA was initially generated by PCR amplification using a biotinylated primer,and was bound with streptavidin to biotinylated polyclonal antibody. Human recombinant p24 antigen sandwiched by antibodies was detected by amplifying the reporter DNA using PCR. The optimal concentration of sreptavidin and DNA label were determined using square titration. The electrophoresis gels were imaged and analyzed by Quantity One software. Results The optimal concentration of sreptavidin and DNA label were determined to be 0.1 mg/ L and 10 ng/ L,respectively. The detection limit of the immuno-PCR assay was 0.1 ng/L,higher than that of the conventional ELISA. Conclusion A highly sensitive immuno-PCR for human recombinant HIV-p24 antigen was indicated to be an available method for early screening of HIV infectors in blood donors.

OBJECTIVE To analyze pathogenic bacterium distribution and antibiotic resistance of our hospital,and provide scientific basis for clinical rational using of antibiotics.METHODS The patients′ clean catch(midstream)(urine) was collected from Jan 2004 to Dec 2005 and cultivated.Antibiotic sensitivity test and adopted by Kirby-Bauer method.RESULTS The pathogenic bacteria mainly consisted of Gram-negatives,among which Escherichia coli was the most frequent,the others in turn were Pseudomonas aeruginosa,Enterobacter cloacae and(Proteus) mirabilis;Enterococcus were the most common among Gram-positives;fungal infection obviously(increased).The bacteria showed different antibiotic resistance rate and multi-drug resistance.CONCLUSIONS It′s very important for making the clinical use of antibiotic more reasonable and controlling drug resistant strains(transmission).

OBJECTIVE To evaluate primarily the detection limit,specificity and reproducibility of immuno-PCR assay on HIV-1 p24 antigen.METHODS We p24 antigen were detected by established immuno-PCR system,and then the detection limit,specificity and reproducibility were discussed.We quantitatively analyzed the nonspecific amplified bands with fluorescence intensity(FI),and a preliminary determination of the lower specific amplification limit was made.RESULTS We taken(x-+3s)FI of nonspecific amplification as the lower limit of specific amplification signal.The detection limit of immuno-PCR assay was 0.1 ng/L.CONCLUSIONS The detection limit,specificity and reproducibility can meet the needs of HIV-1 p24 antigen detection.

Objective To investigate the infection situation of L-form bacteria in urinary calculi from the patients with urolithia-sis in a hospital to provide a scientific basis for postoperative anti-infection and prevention of urinary stone recurrence.Methods The calculi samples in 265 cases of urinary calculi from October to December 2015 were collected and performed the culture of com-mon bacteria and L-form bacteria respectively.Culture of common bacteria and bacterial L-forms.Results Among 265 cases of uri-nary calculi ,8 cases(3% ,8/265) were L-form Bacterial combined with common bacterial infection ,only 7 cases(2.6% ,7/265) were L-form bacterial infection ,80 cases (30.0% ,80/265) were common bacterial infection.15 strains of L-form bacteria were detected and 96 strains of common bacteria were detected.The drug resistance of L-form bacteria was significantly increased compared with common bacteria.Conclusion The positive rate of L-form bacteria culture of urinary calculi is lower than other domestic reports. Adding hypertonic medium for conducting L-form bacterial isolation and culture in the patients with urinary tract infection can re-duce the false negative.

This paper summarized the experience of caring a patient with pancreatic cancer-related depression treated with pancreaticoduodenectomy and postoperative delayed gastric emptying.The nursing included several key points.On the base of collaboration of muhi-disciplinary teams,to strengthen supportive psychological intervention and safety management after admission;to use multimodal analgesia combined with cognitive behavioral therapy to reduce postoperative pain.After the patient was complicated with gastric emptying obstacles,solution-focus mode,sham feeding and nutritional support programs were implemented.Long-term follow-up with physician-nurse collaboration mode was implemented to enhance quality of life of the patient.

OBJECTIVE To investigate the distribution and the drug resistance changes of the pathogenic bacteria isolated from the blood culture specimens collected during the period of 2006-2008.METHODS Blood culture of patients in our hospital was performed by BacT/Alert 240 and the isolated bacteria were identified by API and Microscan and tested for drug resistance against antimicrobial agent by K-B method.A retrospective analysis was made to the blood culture results during the period of 2006-2008 with WHONET 5.4 software.RESULTS Gram-negative rods were the predominant bacteria which caused sepsicemia.The isolated rates of Escherichia coli took the first place during the period of 2006-2008.Klebsiella pneumoniae,Pseudomonas aeruginosa and Staphylococcus aureus(SAU) were also the most important pathogens which caused blood infection.The infection rate of coagulase negative staphylococcus(CNS) and P.aeruginosa had increasing tendency.Imipenem and meropenem were the most effective antibiotics(100%) to E.coli and K.pneumoniae.Then amikacin and cefoxitin also had high susceptibility to E.coli and K.pneumoniae.Furthermore,drug susceptibility in 2008 was higher than that of two years before.All antibacterials had low drug susceptibility to P.aeruginosa,and its drug resistance rate rised obviously.The drug resistance situation of Acinetobacter baumannii was serious.Except imipenem and meropenem had relatively higher susceptibility(20-69.2%),the susceptibility to other antibacterials was lower than 41.7%.Vancomycin,teicoplanin and quinupristin/dalfopristin were the most effective antibiotics to(SAU).CONCLUSIONS The species and drug resistance of the bacteria isolated from blood specimens have changed.More attention should be paid to the detection and surveillance of bacterial resistance in blood culture to promote the rational use of antibiotics.

Objective To screen genes differentially expressed between dermal papilla cells from occipital and vertex scalp of patients with androgenic alopecia (AGA) through a 3D culture system.Methods Dermal papilla cells isolated from the occipital scalp tissue of patients with AGA were cultured in a 2D system for several days.Then,the third-passage dermal papilla cells were subjected to a 3D culture with the presence of dihydrotestosterone (DHT) for 72 hours (experimental group).The dermal papilla cells isolated from the vertex scalp tissue of patients with AGA,which were cultured in a 3D system with dimethyl sulfoxide,but not DHT,served as the control group.Subsequently,total RNA was extracted from the cells and reversely transcribed into cDNA followed by labeling with Cy3 and hybridization to a NimbleGen microarray.Gene ontology (GO) and pathway analysis was carried out to screen differentially expressed genes between the experimental and control group.Real time PCR was conducted to validate the results of microarray analysis.Results As the genome-wide expression profile analysis showed,there were 622 genes differentially expressed between the experimental group and control group,of which,359 were up-regulated and 263 were down-regulated in the experimental group compared with the control group.The above results were corffirmed by real time PCR.GO analysis revealed that the up-regulated genes,such as the CHEK1 and Tobl genes,were mainly involved in the inhibition of cell proliferation and promotion of cell apoptosis,while the down-regulated genes,such as the BAMBI,EFNA3,Dlx3 and UCGC genes,were associated with the acceleration of cell proliferation as well as the growth and development of epidermis.Pathway analysis showed that cell circle-controlling molecules were the most abundant molecules.Conclusions Numerous signalling molecules and pathways are involved in the development of AGA,which are mainly responsible for the modulation of cell circle,proliferation and apoptosis.

Objective To evaluate the effect of family interventions on the prevention of falls in elderly hypertensive patients. Methods One hundred elderly hypertensive patients were divided into the experiment group and the control group in equal number. The control group returned for regular visits after discharge while the experiment group received the family intervention including cognitive,psychological,behavioral and environmental intervention.The two groups were compared in terms of fall rate and degree of injury.Results The incidence of falls in the experiment group was significantly lower than that of the control group,the incidence of soft tissue injury after a fall in the experiment group was significantly lower than that of the control group(both P<0.05).Conclusion Family intervention is effective in prevention of falls in elderly hypertensive patients for it may reduce the incidence of falls and the degree of fall injuries.

Objective To validate the four chronic kidney disease epidemiology collaboration (CKD-EPI) predictive equations based on serum creatinine (SCr) and cystatin C (Cys C) in Chinese CKD patients,and try to develop the GFR predictive equations for Chinese CKD patients.Methods254 CKD patients were randomly selected from four Grade ⅢA hospitals in different regions in China from September 2007 to December 2010.Clearance of dual plasma sampling 99mTc-DTPA was used to measure glomerular filtration rate (rGFR) in 254 CKD patients.The serum concentration of Cr and Cys C were measured.CKD-EPI SCr equation,Cys C equation,Cys C equation adjusted for age,sex and race,SCr/Cys C combinated equation adjusted for age,sex and race were used to estimate GRF ( labeled as eGFR1,eGFR2,eGFR3 and eGFR4,respectively).The correlation,bias and precision of eGFRs were compared with rGFR by Wilcoxon signed rank test,intraclass correlation coefficient (ICC) and Spearman correlation analysis.The deviation degree between rGFR and different eGFRs was compared via Bland-Altman graph.The accuracy within 15％,25％,30％ ( P15,P25,P30) and the staging correctness of eGFR against CKD at different stages was calculated.ResultsThe rGFR in 254 CKD participants was [ 48.07 (26.19 -92.97 )] ml · min -1·(1.73 m2) -1.The Spearman correlatiou coefficients (CC) of eGFR and rGFR varied within the range of 0.873 - 0.896 ( P =0.000 ).The intra-class CC ( ICC ) varied within the range of 0.920 - 0.942.The correlation of eGFR4 was the best.The absolute deviations of 4 eGFRs and deviation precision were eGFR4 ＜eGFR3 ＜ eGFR2 ＜ eGFR1.The 95％ confidence intervals for the regression line of 4 eGFRs shown by Bland-Altman graphs were 92.5,87.3,83.0 and 76.1 ml · min-1 · ( 1.73 m2 ) -1,respectively,with the best result of eGFR4.For P30,the correctness of 4 eGFRs were eGFR4 ＞ eGFR3 ＞ eGFR2 ＞ eGFR1,but no significant difference was found by Chi square test (x2 =6.448,P =0.092).The overall correctness rate in 4 eGFRs against CKD stages were 48.4％ -57.5％,with the highest consistency of eGFR4,but their staging correctnessratewerenotideal(Kappa values were 0.405,0.348,0.366 and 0.463,respectively).Conclusions Compared with CKD-EPI SCr equation,no advantage was found in CKD-EPI Cys C equation.The Cys C equation adjusted by age and sex shows a little advantages over CKD-EPI Cys C equation in bias,precision,correlation and accuracy.The CKD-EPI SCr/Cys C combinated equation adjusted by age,sex and race has advantage over other three equations not only in bias,precision,correlation and accuracy,but also in staging correctness.However,the validation of this equation is still not fairly ideal for Chinese CKD patients.Based on these findings,it is essential for the Chinese CKD patients to develop SCr/Cys C combined predictive equation which adjusted by age,sex or other factors.(Chin J Lab Med,2012,35:798-804)