Antibody Production for a Rapid Fluorescence Polarization Immunoassay of Estrone

Estrone has been identified as a potential endocrine-disrupting chemical （EDC）[1]. Estrone is usually quantified by gas chromatography-mass spectrometry （GC-MS）, GC-MS/MS, high performance liquid chromatography （HPLC）, HPLC- MS, and HPLC-MS/MS, etc.[2-3]. Meanwhile, several immunoassays based on radioimmunoassay, enzyme linked immunosorbent assay （ELISA） or chemiluminescence immunoassay （CLIA） for determination of estrone in real samples have been developed[2＇4]. Although these methods are sensitive, they need multistage separation and are thus time-consuming and laborious. A very promising way for the simplification of immunoassays for routine applications is a shift from heterogeneous methods （with separation） to homogeneous assays （without separation）[5]. Fluorescence polarization immunoassay （FPIA） is one of the homogeneous techniques that meets the requirements of a simple, reliable, fast, and cost-effective analysis[6]. Therefore, the present study is focused on the development of FPIA in order to analyze estrone based on antibody production.

Evaluation of patients with dry eye disease for conjunctival Chlamydia trachomatis and Ureaplasma urealyticum

AIM：To determine the possibility of the development of dry eye disease（DED） as a result of persistent infection with Chlamydia trachomatis and Ureaplasma urealyticum in the conjunctiva of patients.METHODS： This study was conducted on 58 patients of age range 20-50 y,diagnosed with DED confirmed by Schirmer I test and tear breakup time.The non-dry eye control group included 27 subjects of the same age.Ocular specimens were collected as conjunctival scrapings and swabs divided into three groups： the first used for bacterial culture,the second and third taken to detect Chlamydia trachomatis and Ureaplasma urealyticum by direct fluorescent antibody（DFA） assay and polymerase chain reaction（PCR） method. RESULTS： Chlamydia trachomatis was detected in 65.5% and 76% of DED patients by DFA and PCR methods respectively.Ureaplasma urealyticum was found in 44.8% of DED infected patients using the PCR method.Both organisms were identified in only 37.9% of DED patients found to be infected.Control subjects had a 22%detection rate of Chlamydia trachomatis by DFA assay versus a 7% detection rate by PCR; while Ureaplasma urealyticum was detected in 3.7% of the controls by PCR method.The conjunctival culture revealed that gram positive microorganisms represented 75% of isolates with coagulase negative Staphylococci the most common（50%） followed by Staphylococcus aureus（20%）,whereas gram negative microorganisms occurred in 25% of cases,isolating Moraxella spp.as the most frequent organism. CONCLUSION： Our results tend to point out that Chlamydia trachomatis and Ureaplasma urealyticum were detected in a moderate percentage of patients with DED,and could be a fair possibility for its development.PCR is more reliable in detecting Chlamydia trachomatis than DFA technique.The presence of isolated conjunctival bacterial microflora can be of some potential value.

Preconditioning crush increases the survival rate of motor neurons after spinal root avulsion

In a previous study, heat shock protein 27 was persistently upregulated in ventral motor neurons following nerve root avulsion or crush. Here, we examined whether the upregulation of heat shock protein 27 would increase the survival rate of motor neurons. Rats were divided into two groups： an avulsion-only group （avtflsion of the L4 lumbar nerve root only） and a crush-avulsion group （the L4 lumbar nerve root was crushed 1 week prior to the avulsion）. Immunofluores- cent staining revealed that the survival rate of motor neurons was significantly greater in the crush-avulsion group than in the avulsion-only group, and this difference remained for at least 5 weeks after avulsion. The higher neuronal survival rate may be explained by the upregulation of heat shock protein 27 expression in motor neurons in the crush-avulsion group. Further- more, preconditioning crush greatly attenuated the expression of nitric oxide synthase in the motor neurons. Our findings indicate that the neuroprotective action of preconditioning crush is mediated through the upregulation of heat shock protein 27 expression and the attenuation of neuronal nitric oxide synthase upregulation following avulsion.

Diagnosis,fetal risk and treatment of pemphigoid gestationis in pregnancy:A case report

BACKGROUND Pemphigoid gestationis(PG)is a rare autoimmune blistering disease that usually presents in the second or third trimester,with an incidence of 1 per 50000 pregnancies.PG tends to recur with an earlier onset and a more severe course in subsequent pregnancies.Skin biopsy markers can be confirmed by direct immunofluorescence staining.CASE SUMMARY Our patient was diagnosed with PG at 8 mo of gestation with fresh bullous lesion marks on the abdomen and limbs.Termination of the pregnancy was performed by cesarean section at 37+4 wk of gestation.The patient delivered an infant weighing 3620 gm.The infant had urticaria-like and vesicular skin lesions and was diagnosed with PG.The patient was discharged on prednisolone and in a satisfactory condition.The infant was discharged after anti-inflammatory therapy for one week.CONCLUSION PG is a rarely reported disease,and 10%of newborns develop mild clinical symptoms consisting of urticaria-like or vesicular skin lesions.We intend to remind clinicians to consider this condition when a patient presents with such lesions so that treatment can be started early and neonatal morbidity can be taken into account.

Detection of Epstein-Barr virus infection subtype in patients with multiple sclerosis by indirect immunofluorescence assay

Aim:The aim was to investigate the infectious conditions of Epstein-Barr virus(EBV)in patients with multiple sclerosis(MS).Methods:Cerebrospinal fluid(CSF)of 20 patients with MS and 20 with other neurological diseases(OND)were tested with indirect immunofluorescence for anti-EBV capsid antigen(EBV-CA)immunoglobulin G(IgG),IgG affinity for anti-EBV-CA,anti-EBV-CA immunoglobulin M(IgM),anti-EBV early antigen(EBV-EA)IgG and anti-EBV nuclear antigen(EBNA)IgG.According to the pattern of antibodies in CSF,infection rates of acute,chronic,primary,recurrent,and past infections were analyzed in the two groups of patients.Results:There were no significant differences in anti-EBV-CA,anti-EBC-EA,and anti-EBNA antigen IgG in CSF between MS and OND patients(P>0.05).The positive rate of low affinity for anti-EBV-CA IgG in MS patients was significantly higher than that for OND patients(75%vs.40%,P<0.05).Furthermore,significant differences in the positive rate of anti-EBV-CA IgM were found between MS and OND patients(70%vs.25%,P<0.05).Of the MS patients,75%were in an EBV acute infection state compared with 40%of OND patients(P<0.05).Conclusion:Acute infection of EBV closely correlates with the occurrence of MS.