Region and layer-specific expression of GABAA receptor isoforms and KCC2 in developing cortex.

Introduction: γ-Aminobutyric acid (GABA) type A receptors (GABAARs) are ligand-gated Cl-channels that mediate the bulk of inhibitory neurotransmission in the mature CNS and are targets of many drugs. During cortical development, GABAAR-mediated signals are significantly modulated by changing subunit composition and expression of Cl-transporters as part of developmental processes and early network activity. To date, this developmental evolution has remained understudied, particularly at the level of cortical layer-specific changes. In this study, we characterized the expression of nine major GABAAR subunits and K-Cl transporter 2 (KCC2) in mouse somatosensory cortex from embryonic development to postweaning maturity. Methods: We evaluated expression of α1-5, ß2-3, γ2, and δ GABAAR subunits using immunohistochemistry and Western blot techniques, and expression of KCC2 using immunohistochemistry in cortices from E13.5 to P25 mice. Results: We found that embryonic cortex expresses mainly α3, α5, ß3, and γ2, while expression of α1, α2, α4, ß2, δ, and KCC2 begins at later points in development; however, many patterns of nuanced expression can be found in specific lamina, cortical regions, and cells and structures. Discussion: While the general pattern of expression of each subunit and KCC2 is similar to previous studies, we found a number of unique temporal, regional, and laminar patterns that were previously unknown. These findings provide much needed knowledge of the intricate developmental evolution in GABAAR composition and KCC2 expression to accommodate developmental signals that transition to mature neurotransmission.

Search for antibodies against Trichinella in two synanthropic Procyonidae species from southeast Mexico: white-nosed coatis (Nasua narica) and raccoons (Procyon lotor).

Trichinella is a nematode that are spread by the consumption of parasitized meat. Carnivora, a mammalian order, serve as key hosts for this parasite. However, evidence of Trichinella in wildlife from the Neotropics is extremely scarce, with reports documenting its presence only for five carnivore species: two Felidae, one Otariidae and two Mustelidae. Other widely distributed species that are consumed as bushmeat, such as Procyonidae, have not been studied in this context. A long-term study was performed for antibodies against Trichinella in coatis (Nasua narica) and common raccoons (Procyon lotor) in southeastern Mexico. Between the summer of 2009 to the winter 2013, a total of 291 coati samples and 125 raccoon samples were collected from a tropical green area located within an urban zone. An Enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies against the excretory and secretory products of Trichinella spiralis muscle larva. ELISA-positive samples were further confirmed by Western Blot analysis. Results showed no evidence of antibodies during the first two years of study. However, in 2011, a sudden appearance of anti-Trichinella occurred. The seroprevalence reached its highest peak of 43% for coatis during winter 2013 and 53% for raccoons in summer 2013. This is the first study that provides evidence of Trichinella circulation within a neotropical procyonid community.

Bone microenvironment regulative hydrogels with ROS scavenging and prolonged oxygen-generating for enhancing bone repair.

Large bone defects resulting from fractures and disease are a major clinical challenge, being often unable to heal spontaneously by the body's repair mechanisms. Lines of evidence have shown that hypoxia-induced overproduction of ROS in bone defect region has a major impact on delaying bone regeneration. However, replenishing excess oxygen in a short time cause high oxygen tension that affect the activity of osteoblast precursor cells. Therefore, reasonably restoring the hypoxic condition of bone microenvironment is essential for facilitating bone repair. Herein, we designed ROS scavenging and responsive prolonged oxygen-generating hydrogels (CPP-L/GelMA) as a "bone microenvironment regulative hydrogel" to reverse the hypoxic microenvironment in bone defects region. CPP-L/GelMA hydrogels comprises an antioxidant enzyme catalase (CAT) and ROS-responsive oxygen-releasing nanoparticles (PFC@PLGA/PPS) co-loaded liposome (CCP-L) and GelMA hydrogels. Under hypoxic condition, CPP-L/GelMA can release CAT for degrading hydrogen peroxide to generate oxygen and be triggered by superfluous ROS to continuously release the oxygen for more than 2 weeks. The prolonged oxygen enriched microenvironment generated by CPP-L/GelMA hydrogel significantly enhanced angiogenesis and osteogenesis while inhibited osteoclastogenesis. Finally, CPP-L/GelMA showed excellent bone regeneration effect in a mice skull defect model through the Nrf2-BMAL1-autophagy pathway. Hence, CPP-L/GelMA, as a bone microenvironment regulative hydrogel for bone tissue respiration, can effectively scavenge ROS and provide prolonged oxygen supply according to the demand in bone defect region, possessing of great clinical therapeutic potential.

False positive reaction of COVID-19 antibody ELISA screening: internal logic of various methods / 中国输血杂志

【Objective】 To analyze the results of different methods for reactive samples screened by the enzyme linked immunosorbent assay (ELISA) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in blood donors. 【Methods】 From March to April 2020, a total of 8 632 blood samples in Shenzhen were screened for SARS-CoV-2 total antibodies (TAb, including IgG, IgM, IgA) in plasma using ELISA(PC group), the antibody reactivity samples and their follow up plasma samples (FC group), and samples of disease control group(DC group) from January to April 2020 were detected using the following methods: 1) ELISA method for detecting IgG, IgM, and (or without detection) TAb; 2) pseudovirus neutralizing antibody test(pVNT); 3) western blot (WB) of SARS-CoV-2 antibody. The negative control group(NC group) from February to April 2020 performed ELISA and WB testing. 【Results】 Among the 34 total antibody positive samples, 2 were positive for pVNT test, and the total antibody, IgG and WB in the initial screening and tracking testing were positive. Thereafter, it was determined to be confirmed positive. The other 2 cases were positive for pVNT test, while the samples with positive WB results were in the follow-up stage. The TAb, IgG, and pVNT results did not conform to the dynamic evolution of antibodies, and cannot be determined as confirmed positive. 【Conclusion】 The infection status of antibody reactivity samples screened by SARS-CoV-2 ELISA can be judged by the logic of pVNT, WB and the dynamic change of antibody.

Urine-Derived Stem Cells Express 571 Neuromuscular Disorders Causing Genes, Making Them a Potential in vitro Model for Rare Genetic Diseases.

Background: Neuromuscular disorders (NMDs) are a heterogeneous group of genetic diseases, caused by mutations in genes involved in spinal cord, peripheral nerve, neuromuscular junction, and muscle functions. To advance the knowledge of the pathological mechanisms underlying NMDs and to eventually identify new potential drugs paving the way for personalized medicine, limitations regarding the availability of neuromuscular disease-related biological samples, rarely accessible from patients, are a major challenge. Aim: We characterized urinary stem cells (USCs) by in-depth transcriptome and protein profiling to evaluate whether this easily accessible source of patient-derived cells is suitable to study neuromuscular genetic diseases, focusing especially on those currently involved in clinical trials. Methods: The global transcriptomics of either native or MyoD transformed USCs obtained from control individuals was performed by RNA-seq. The expression of 610 genes belonging to 16 groups of disorders (http://www.musclegenetable.fr/) whose mutations cause neuromuscular diseases, was investigated on the RNA-seq output. In addition, protein expression of 11 genes related to NMDs including COL6A, EMD, LMNA, SMN, UBA1, DYNC1H1, SOD1, C9orf72, DYSF, DAG1, and HTT was analyzed in native USCs by immunofluorescence and/or Western blot (WB). Results: RNA-seq profile of control USCs shows that 571 out of 610 genes known to be involved in NMDs, are expressed in USCs. Interestingly, the expression levels of the majority of NMD genes remain unmodified following USCs MyoD transformation. Most genes involved in the pathogenesis of all 16 groups of NMDs are well represented except for channelopathies and malignant hyperthermia related genes. All tested proteins showed high expression values, suggesting consistency between transcription and protein representation in USCs. Conclusion: Our data suggest that USCs are human cells, obtainable by non-invasive means, which might be used as a patient-specific cell model to study neuromuscular disease-causing genes and that they can be likely adopted for a variety of in vitro functional studies such as mutation characterization, pathway identification, and drug screening.

A retrospective analysis of 2 728 HIV antibody screening positive samples / 公共卫生与预防医学

Objective To analyze the positive results of HIV antibody screening in the laboratory of AIDS confirmation center of Hubei Provincial Center for Disease Control and Prevention from 2014 to 2020, and to provide a basis for improving detection strategies. Methods A total of 2 728 primary screening positive specimens received by the laboratory of Hubei confirmation center from 2014 to 2020 were retested with two reagents. Specimens with at least one reactive result were confirmed with western blot (WB). The samples with uncertain or negative WB results were further confirmed by nucleic acid quantitative detection. The test results were analyzed retrospectively. Results A total of 2 297 specimens with positive retest results were confirmed by WB, with a positive rate of 93.47%. The highest proportion of patients was from medical institutions. The positive rate detected by 4 diagnostic kits was apparently higher in S/CO＞10 cases than that in S/CO≤10, and the difference was statistically significant (P 5 000cps / ml, and 12 cases were TND. 13 of the 30 WB negative samples had nucleic acid test results＞5 000CPs/mL . Conclusions The coverage of HIV screening laboratories in hospitals at all levels should be further increased to find more HIV infected persons. The anti-HIV ELISA S/CO ratio is correlated with the positive results confirmed by western blot. Therefore, ELISA S/CO ratio can be used to predict anti-HIV antibody positivity. For samples with uncertain or negative WB detection, supplemental nucleic acid test should be carried out timely for early diagnosis.

The association of NF2 (neurofibromin 2) gene polymorphism and the risk of medulloblastomas.

To explore the relationship between NF2 promoter gene mutation and the risk of medulloblastomas (MBs). We collected tissues from 16 MB patients and 7 age-matched non-MB controls. Gene sequencing, qPCR (real-time quantitative polymerase chain reaction), IHC (immunohistochemistry), and WB (Western blot) were used to analyze the changes in the NF2 gene sequence and expression between patients and controls. We found that NF2 promoter gene mutations occurred in MB patients. The NF2 mRNA expression was higher in the controls than in patients (p = 0.03 < 0.05); however, the results of IHC and WB demonstrated that the NF2 protein expression was significantly higher in patients than in the controls (IHC: p = 0.0001; WB: p = 0.01). There was no significant difference in the CRL4 mRNA and protein levels. In addition, NF2 protein was mainly expressed in the nucleus in MB patients, while the NF2 protein was mainly expressed in the cytoplasm in the controls. NF2 promoter mutations exist in MB patients. NF2 mRNA expression was higher in controls than patients; whereas NF2 protein level was higher in patients than in controls.

Comparing the differences between domestic and imported EIA-HIV screening results for voluntary blood donors / 中国输血杂志

Objective To compare the differences of two kinds of EIA reagents for HIV-1/2 (Ab/Ag) screening results of voluntary blood donors,in addition to find out the feasibility of reducing 1 times of EIA detection.Methods To collect data of HIV 1/2 screening positive results and confirmatory test for voluntary blood donors from 2009 to 2014 in Jiaxing area,and to compare the relationship of screeing test results with that of the confirmatory test,and then to analyze the relevance between S/CO values of screening test and confirmatory test.Results Screening positive rates of domestic and imported reagents,which were 9.58/10 000 and 12.43/10 000,respectively;and the confirmatory coincidence rates were 11.84％ and 9.12％,respectively.There was no significant difference (x2 =1.11,P＞0.05).The double-reagent joint detection positive rate was 1.37/10 000,and its positive predictive value was 82.86％.Single-reagent test result compared with that of double-reagent test,which had significant differences (x2domestic =94.04,P＜0.05 and x2ximported =124.86,P＜0.05).When the S/CO value was more than 6,domestic and imported reagents positive predictive values were 93.55％ (29/31) and 87.50％ (28/32),respectively.Conclusion There is no difference between domestic and imported reagents EIA-HIV1/2.

Sero-epidemiological value of some hydatid cyst antigen in diagnosis of human cystic echinococcosis.

Cystic echinococcosis (CE) is a severe zoonotic disease which affects both human and animals. The disease has a considerable economic and social impact, because it has numerous complications leading to important disabilities and even death. CE is a widespread chronic endemic helminthic disease caused by infection with metacestodes of tapeworm Echinococcus granulosus. This study was conducted to diagnosis human CE by hydatid cyst antigens from camels and sheep. Hydatid fluid and protoscoleces crude antigens corresponding to camel and sheep were resolute by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions and the protein bands of different antigens were exposed to infected patients serum CE through western blot (WB) assay. The camel hydatid fluid antigen revealed five polypeptide bands of 18-98.8 kDa by SDS-PAGE while sheep hydatid fluid antigen revealed four polypeptide bands of 20-100 kDa. Immune reactive bands were obtained through WB ranged from 25 to 125 kDa. The study showed prominent immune reactive bands of 92, 52.2 and 35.7 kDa which may helpful in diagnosis of human CE.

High rate of missed HIV infections in individuals with indeterminate or negative HIV western blots based on current HIV testing algorithm in China.

It remains unclear if China's current HIV antibody testing algorithm misses a substantial number of HIV infected individuals. Of 196 specimens with indeterminate or negative results on HIV western blot (WB) retrospectively examined by HIV-1 nucleic acid test (NAT), 67.57% (75/111) of indeterminate WB samples, and 16.47% (14/85) of negative WB samples were identified as NAT positive. HIV-1 loads in negative WB samples were significantly higher than those in indeterminate WB samples. Notably, 86.67% (13/15) of samples with negative WB and double positive immunoassay results were NAT positive. The rate of HIV-1 infections missed by China's current HIV testing algorithm is unacceptably high. Thus, China should consider using NAT or integrating fourth generation ELISA into current only antibodies-based HIV confirmation. J. Med. Virol. 88:1462-1466, 2016. © 2016 Wiley Periodicals, Inc.

Diagnóstico de infecção por vírus linfotrópicos de células T humanas dos tipos 1 (HTLV-1) e -2 (HTLV-2) em população de risco: passado, presente e futuro / Diagnosis of human T-cell lymphotropic virus type 1 (HTLV-1) and type 2(HTLV-2) infection in at-risk populations: past, present and future

O Brasil é o país com o maior número de pessoas infectadas pelos vírus linfotrópicos de células T humanas dos tipos 1e -2 (HTLV-1 e HTLV–2) com mais de 2,5 milhões de indivíduos infectados. Em 1993, a realização de testes sorológicos específicos tornou-se obrigatória em Bancos de Sangue. O HTLV-1 causa leucemia/linfoma de células T do adulto e mielopatia associada ao HTLV-1/paraparesia espástica tropical além de outras doenças, enquanto o HTLV-2 pode causar alguns quadros neurológicos e alterar a evolução de HIV/Aids. Os testes sorológicos que identificam anticorpos específicos disponíveis no mercado têm falhado no diagnóstico, principalmente de infecção por HTLV-2. Vários algoritmos de testes de triagem e confirmatórios têm sido propostos, mas nenhum deles se mostrou 100% eficiente com casuística de alto risco. Muitos soros resultam em padrão indeterminado no Western blot, e os isolados virais utilizados na composição dos kits podem ser a causa desses resultados. As técnicas de biologia molecular têm sido descritas como testes confirmatórios, mas não têm sido empregadas na rotina. Desde 1991, a Seção de Imunologia do Instituto Adolfo Lutz tem estudado a infecção por HTLV-1/2, contribuindo para o diagnóstico sorológico e molecular, e tem como desafio implantar um teste laboratorial capaz de detectar infecção causada por cepas brasileiras de HTLV-2.