Objective To evaluate the quality of four domestic and three imported fourth-generation HIV diagnostic reagents.Methods The specificity and sensitivity of these assays were analyzed when testing HIV negative samples and HIV-1 RNA positive samples.The relative seroconversion sensitivity index was analyzed when testing BBI seroconversion panels.Results The sensitivity of seven 4th-generation assays were 100％ (95％ CI:99.86％-100％ ),and one sample at the window period of HIV-1 infection were detected as positive.Of the seven assays,one imported assay exhibited the relative largeδ + value (1.0892),and the small δ+ value were found on the remaining six assays (0.0836-0.3003 ).For the samples negative for HIV antibody,varying degrees of false positives were observed on the seven assays ( specificity:97.80％ -99.60％,δ- value:-1.3803 to -0.4778).When testing the BBI seroconversion panels,the relative seroconversion sensitivity index of domestic assays were -0.500-0,however,which of imported assays were -0.600 and -0.700.Conclusion The seven reagents exhibited high sensitivity and specificity.The 4th generation HIV assays can be used as blood screening reagents to find the samples at window period of HIV-1 infection,thus indicating the certain meaning in reducing the transmission risk of HIV-1 for fourth-generation HIV diagnostic reagents.However,the better efficiency to detect HIV-1 early infection was observed on the imported assays than on the domestic assays.

Object To investigate the feasibility that Rhizoma Dioscoreae in different areas was identified by thermal analysis Methods By comparison of the figures for the character of the DTA and DTG curves in N 2 and air environment, Rhizoma Dioscoreae from five areas was identified Results The DTA and DTG curves of varied Rhizoma Dioscoreae were different from each other Conclusion The method can be used in the identification of a variety of Rhizoma Dioscoreae from different areas

Objective To evaluate the differences between the third and the fourth generations of anti-HIV assays,and different kits within the same generation.Methods A total of 989 HIV-negative samples,185 samples positive for HIV-1 RNA.1st-generation international references of HIV antibodies and samples from 9 sets of BBI seroconversion panels were detected by 8 kits of the third generation and 4 kits of the fourth.Results The fourth generation kits can detect HIV infection earlier than the third generation kits.However,the detected days of HIV infection with different kits of the fourth generation were different whilst no significant difierences were found with difierent kits of the third generation.Furthermore,the capacity of detecting samples with different genotypes for different reagents was different,especially the capacity of domestic reagents on detecting HIV-1 O group and HIV-2 samples was relatively weak.Conclusion These data provided information to improve the quality of anti-HIV diagnostic reagents further.

Objective To explore influences of improved intramuscular injection on quality of benzathine benzylpenicillin medication.Methods The cluster random sampling was adopted to select 178 patients who needed injection of 240U benzathine benzylpenicillin.A self-control study design was used,and benzathine benzylpenicillin was injected in both sides with each of 1 200 000 units.The left side was injected via routine method,while the right side was injected by an improved intramuscular injection.One-time success rate,degree and duration of pain during and after injection were recorded.Results The differences of one-time success rate,pain during injection,pain after injection,duration of pain after injection and incidence of induration after injection between two groups were statistically significant(P＜0.01).Conclusion The improved intramuscular injection can improve one-time success rate of benzathine benzylpenicillin,reduce pain during injection and local pain after injection,shorten duration of pain and decrease incidence of induration after injection.

AIM: To prepare and purify the polyclonal antibodies against human myofibrillogenesis regulator 1 (hMR-1), then to characterize the purity, titer, specificity and the availability.METHODS: Two polypeptides named peptide 1 and 2 were synthesized based on the bioinformatics analysis of the sequence of hMR-1 by using software TMHMM and DNAStar, then coupled with keyhole limpet hemocyanin (KLH) for immunization. These peptides for immunization were mixed and injected into New Zealand rabbits to prepare antibodies specifically against hMR-1. ELISA assay was used to detect the titers of the antibodies. After purification by immunoaffinity chromatography, antibodies were identified by Western blotting and immunocytofluorescent assays. Applications of the antibodies on neonatal rat cardiomyocytes were also employed.RESULTS: (1)The titers of antibodies were 1:10~5. In WB assay, a specific 17kD band was detected, corresponding to the predicted molecular weight of hMR-1; the positive fluorescent signals were distinct. (2)On the neonatal rat cardiomyocytes model, we observed a peri-nucleus location. The fluorescent signal of hMR-1 overexpression group was much stronger than that in vector control and normal control groups.CONCLUSION: All these results indicate that the antibodies obtained from poly peptides mixture immunization have either human original or rat original antigens. The antibody is available for using in Western blotting or immunofluorescent assays.

Objective To investigate the pathogens and their drug resistance of neonatal skin infection in Maternal-Neonatal Unit to provide evidence for rational selection of antibiotics.Methods Secretions of skin lesions from newborns with skin infections delivered in Maternal-Neonatal Unit,Department of Obstetrics of Beijing Daxing People's Hospital from Jan.2009 to Dec.2010 were collected,cultured and identified for bacterial serotype.Antibiotic resistances of pathogens were determined.Enumeration data was presented by frequency and rate.The difference between groups was compared with Chi-square test.Results Two hundred and six newborn skin infection cases were diagnosed,which accounted for 2.3％ of all newborns (n=9131) delivered in the hospital during the study period.Two types of skin infection were identified,impetigo (n=192,93.2％) and omphalitis (n=14,6.8％).Totally 154 pathogens were found,including 95 (61.7％) Gram-positive cocci and 59 (38.3 ％) Gram-negative bacilli.The major pathogens among Gram-positive cocci were Staphylococcus aureus (45/154, 29.2％), followed by Staphylococcus epidermidis and Staphylococcus haemolyticus.Staphylococcus was sensitive to Cefazolin,Amoxicillin/clavulanic acid and Piperacillin/tazobactam,while resistant to Penicillin,Ampicillin and Erythromycin.The major pathogens among Gram-negative bacilli were Enterobacter cloacae (19/154,12.3％),followed by Escherichia coli and Klebsiella pneumoniae.Drug resistance rate of Gram-negative bacilli was high in penicillin,aminoglycosides and cephalosporins,while they were sensitive to piperacillin/tazobactam,Ampicillin/sulbactam,carbapenems and Quinolones.Conclusions Gram-positive cocci are the main pathogens of neonatal skin infection in the Maternal-Neonatal Unit,and among which Staphylococcus aureus is the major pathogen.Multi-drug resistance is common in the identified pathogens.Attentions should be paid to use antibiotics reasonably according to drug sensitivity test.

Objective To explore neurobiological mechanisms of the withdrawal-induced aversion,the changes of protein kinase A(PKA) were measured in shell of accumbens nucleus (AcbSH) of CPA model rats.Methods 1.All 36 male SD rats were divided into three groups,model group ( MN group),and control group (MS group and SN group).MN group was injected with morphine,6.5 days,10mg/kg,intraperitoneally (IP),twice per day,naloxone injection,0.3 mg/kg,ip,along with conditioned place aversion training,to develop the CPA model.The MS group was administrated equivalent volume of morphine and saline.Also the SN group was injected with equivalent volume of saline and naloxone.2.During the development of CPA,the expression of protein kinase A was assayed with immunohistochemistry in the AcbSH.Results Before the development of CPA,PKA expressions were no significant differences among the three groups in the AcbSH (F=2.306,P=0.130).However,after development of CPA,PKA expressions showed significant differences among the three groups(F =36.516,P =0.000).The average gray intensity of MN group (109.50 ± 4.661 ) was apparently higher than the MS group (126.50 ±3.697,P＜0.01),than the SN group (133.50 ±6.364,P＜0.01).Conclusions 1.Protein kinase A expression,leading to the aversion in the AcbSH probably is a key pathway contributing to the development of CPA.2.The neuroadaptation mediated by PKA may be one of important molecular underpinnings of CPA.

Swabbing samples were routinely collected from health care workers ( HCW) contacting newborns in Obstetrics Department of our hospital on every Wednesday during January to December 2015. Total 495 samples from nasal vestibule and 483 samples from hands of HCW were collected for bacterial culture.The carrying status of methicillin-resistant staphylococccus aureus ( MRSA) was analyzed.The results showed that the average carrying rate of MRSA in the nasal vestibule was 8.1%(40/495) and that in hands was 5.0(24/483)(χ2 =3.871, P<0.05).The MRSA detection rate of nasal vestibular samples in the second quarter was the highest [12.2%(17/139)], while that of hands samples was highest in the first quarter [ 15.3%( 15/98 ) ] . The study indicates that carrying rate of MRSA in health care workers contacting newborns of our hospital is high , and the training and monitoring should be strengthened among health workers to reduce the MRSA carrying rate .

Objective To pan and characterize anti-HIV-1 Fab by the phage antibody library technology. Methods Total RNA were extracted from lymphocytes which were isolated from peripheral blood collected from asymptomatic HIV-1 infected donors with high titer antibody against HIV-1. The genes of heavy chains Fd fragment and light chains of antibody were amplified by RT-PCR. The phagmids pComb3X cloned Fd and light chain genes were transformed into E. coli XL1-Blue by electroporation to construct phage Fab library. By three runs of "absorption-elution-neutralization-enrichment", the clones were induced by IPTG and characterized by ELISA. The positive clones were sequenced and analyzed the sequences. Subsequently, Fab antibodies of these positive clones were induced to expressed and purified, then the recombinant virus neutralization assay was performed. Results A phage Fab library was constructed with 8×106 members, and 11 positive clones were obtained by detecting IPTG-induced-expressing Fabs with ELISA. By analysis of the sequences, 10 light chain genes and 8 Fd genes were ensured to be obtained. Compared with the genes of anti-HIV-1 antibodies in HIV sequence database, the gene sequences we obtained were highly homologous to some patent genes of anti-HIV-1 gp120 antibodies in HIV sequence database( light chains with 60%-90% identity, Fd with 71%-85% identity); The CDRs of these positive clones were determined by comparing the positive clone genes with antibodies' genes in V base database, furthermore, CDRH3 of these positive clones has the length of 12-22 aa. Strand shift had little effect to improving affinity of our Fab clones. Fab antibodies were induced to express at the concentration of ＞ 10 mg/L. Three Fab antibodies neutralize HIV-1 virus to some extent. Conclusion The studies will provide the basis on further study on the anti-HIV-1 Fabs obtained successfully.

Objective To investigate the epidemiological and clinical characteristics and prognosis of 31 children with hand foot and mouth disease(HFMD) complicated with acute flaccid paralysis(AFP).Methods Thirty-one HFMD children complicated with AFP served as study objective,who were admitted to Tangshan Maternity and Children Health Hospital from Jun 2011 to Dec 2014.The clinical manifestations and follow-up outcomes of 31 children with HFMD followed by AFP were retrospectively reviewed.Results Participants in this study consisted of 31 children(16 males,15 females,age from 3 months to 9 years) who met the criteria for HFMD with AFP.Among the 31 cases,27(87.1％)cases were less than 3 years old and 29(93.5％) cases were from countryside.Eleven cases (35.5％)were infected by enterovirus 71,1 case (3.2％) was infected with coxsackieviruses A16 and 19 cases(61.3％) were infected by other enteroviruses.AFP developed(7.1 ±2.9) days after the onset of fever and progressed to maximum severity within 1-2 days.All of the cases presented with fever and skin rash,38.7％ cases presented with limbs tremor and 87.1％ cases presented with startle and skip.Besides AFP of limbs, 100％ cases complicated with encephalitis.Thirty-one cases showed poliomyelitis-like syndrome(20 cases with one flaccid limb,5 cases with lower limbs,4 cases with upper limbs and 2 cases with left hemiplegia).In these cases, the muscle power varied from level 0 to level 4.The muscle strengthen and muscle tone in 21 patients were recovered within 2 weeks,and the other cases showed recovery 2 to 3 weeks later.After four weeks, the muscle strengthen in 19 cases recovered to 5 level(10 cases with single lower limb,5 cases with single upper limb,3 cases with two lower limbs and 1 case with two upper limbs).Conclusion HFMD complicated with AFP most commonly occurs in children aged less than 3 years old.The majority of cases were from countryside.Enterovirus 71 is still relatively common pathogen.All the 31 cases complicated with encephalitis.Cases clinically presented one flaccid limb, lower limbs, upper limbs and hemiplegia.AFP may be to some degree reversible in HFMD cases.In most cases described here, paralysis occurred in a single lower extremity and recovered more rapidly than those with two fimbs affected or with single upper extremity impairment.