Strength-Ductility Mechanism of CoCrFeMnNi High-Entropy Alloys with Inverse Gradient-Grained Structures.

The microstructures and mechanical properties of equiatomic CoCrFeMnNi high-entropy alloys (HEAs) treated with various processing parameters of laser surface heat treatment are studied in this paper. The typical inverse gradient-grained structure, which is composed of a hard central layer and a soft surface layer, can be obtained by laser surface heat treatment. A much narrower gradient layer leads to the highest yield strength by sacrificing ductility when the surface temperature of the laser-irradiated region remains at ~850 °C, whereas the fully recrystallized microstructure, which exists from the top surface layer to the ~1.05 mm depth layer, increases the ductility but decreases the yield strength as the maximum heating temperature rises to ~1050 °C. Significantly, the superior strength-ductility combination can be acquired by controlling the surface temperature of a laser-irradiated surface at ~1000 °C with a scanning speed of ~4 mm/s due to the effect of hetero-deformation-induced strengthening and hardening, as well as the enhanced interaction between dislocation and nanotwins by the hierarchical nanotwins. Therefore, retaining the partial recrystallized microstructure with a relatively high microhardness in the central layer, promoting the generation of hierarchical nanotwins, and increasing the volume proportion of gradient layer can effectively facilitate the inverse gradient-grained CoCrFeMnNi HEAs to exhibit a desirable strength-ductility synergy.

Complete genome sequence of the novel Antarctic Oceanisphaera sp. IT1-181 that carried five plasmids.

Bacteria of the genus Oceanisphaera in the class Gammaproteobacteria are widely distributed in marine environments. Oceanisphaera sp. IT1-181 was isolated from intertidal sediment in the coastal region of the Chinese Great Wall Station on the Fildes Peninsula, King George Island, Antarctica. Here, we sequenced the complete genome of strain IT1-181, which contained a single chromosome of 3,572,184 bp (G + C content of 49.89 mol%) with five plasmids. A total of 3229 protein-coding genes, 88 tRNA genes, and 25 rRNA genes were obtained. Genome sequence analysis revealed that strain IT1-181 was not only a potentially novel species of the genus Oceanisphaera, but also harbored genes involved in biosynthesizing ectoine as well as poly-ß-hydroxybutyric acid (PHB). In addition, genes of a complete type I-E CRISPR-Cas system were found in the bacterium. The results indicate the potential of strain Oceanisphaera sp. IT1-181 in biotechnology and are helpful for us understanding its ecological roles in the changing Antarctic intertidal zone environment.

Investigation on muscle fiber types and meat quality and estimation of their heritability and correlation coefficients with each other in four pig populations.

The aim of this study was to investigate the muscle fiber types and meat quality in four populations and estimate the heritability and correlation coefficients of those traits in Shanxia long black pig (SX). In this study, a total of 318 pigs were recorded for 16 traits of the muscle fiber types and meat quality in four populations, including 256 individuals from the new breed SX. The population had a significant effect on all recorded traits, and the meat quality of the Lulai black pig was better than the remaining populations. The heritability (h2 ) of meat quality traits was from 0.06 (pH at 24 h) to 0.47 (shearing force), and the muscle fiber types belonged to the traits with low to medium heritability. The density of total fiber had the highest h2 (0.40), while the percentage of type IIA had the lowest h2 (0.04). Most traits are phenotypically correlated with each other, but only a small proportion of traits are genetically correlated with each other. None fiber type genetically correlated with meat quality significantly, because the genetic correlation coefficients had large standard errors. These results provided some insights into genetic improvements for the meat quality in pig breeds and also indicated that the parameters of muscle fiber characteristics can explain parts of the variation in meat quality.

Comparative genomic analysis of two Arctic Pseudomonas strains reveals insights into the aerobic denitrification in cold environments.

BACKGROUND: Biological denitrification has been commonly adopted for the removal of nitrogen from sewage effluents. However, due to the low temperature during winter, microorganisms in the wastewater biological treatment unit usually encounter problems such as slow cell growth and low enzymatic efficiency. Hence, the isolation and screening of cold-tolerant aerobic denitrifying bacteria (ADB) have recently drawn attention. In our previous study, two Pseudomonas strains PMCC200344 and PMCC200367 isolated from Arctic soil demonstrated strong denitrification ability at low temperatures. The two Arctic strains show potential for biological nitrogen removal from sewage in cold environments. However, the genome sequences of these two organisms have not been reported thus far. RESULTS: Here, the basic characteristics and genetic diversity of strains PMCC200344 and PMCC200367 were described, together with the complete genomes and comparative genomic results. The genome of Pseudomonas sp. PMCC200344 was composed of a circular chromosome of 6,478,166 bp with a G + C content of 58.60% and contained a total of 5,853 genes. The genome of Pseudomonas sp. PMCC200367 was composed of a circular chromosome of 6,360,061 bp with a G + C content of 58.68% and contained 5,801 genes. Not only prophages but also genomic islands were identified in the two Pseudomonas strains. No plasmids were observed. All genes of a complete set of denitrification pathways as well as various putative cold adaptation and heavy metal resistance genes in the genomes were identified and analyzed. These genes were usually detected on genomic islands in bacterial genomes. CONCLUSIONS: These analytical results provide insights into the genomic basis of microbial denitrification in cold environments, indicating the potential of Arctic Pseudomonas strains in nitrogen removal from sewage effluents at low temperatures.

Caffeine Synthesis and Its Mechanism and Application by Microbial Degradation, A Review.

Caffeine is a metabolite derived from purine nucleotides, typically accounting for 2-5% of the dry weight of tea and 1-2% of the dry weight of coffee. In the tea and coffee plants, the main synthesis pathway of caffeine is a four-step sequence consisting of three methylation reactions and one nucleosidase reaction using xanthine as a precursor. In bacteria, caffeine degradation occurs mainly through the pathways of N-demethylation and C-8 oxidation. However, a study fully and systematically summarizing the metabolism and application of caffeine in microorganisms has not been established elsewhere. In the present study, we provide a review of the biosynthesis, microbial degradation, gene expression, and application of caffeine microbial degradation. The present review aims to further elaborate the mechanism of caffeine metabolism by microorganisms and explore the development prospects in this field.

Peripheral nerve defects repaired with autogenous vein grafts filled with platelet-rich plasma and active nerve microtissues and evaluated by novel multimodal ultrasound techniques.

BACKGROUND: Developing biocompatible nerve conduits that accelerate peripheral nerve regeneration, lengthening and functional recovery remains a challenge. The combined application of nerve microtissues and platelet-rich plasma (PRP) provides abundant Schwann cells (SCs) and various natural growth factors and can compensate for the deficiency of SCs in the nerve bridge, as well as the limitations of applying a single type of growth factor. Multimodal ultrasound evaluation can provide additional information on the stiffness and microvascular flow perfusion of the tissue. This study was designed to investigate the effectiveness of a novel tissue-engineered nerve graft composed of an autogenous vein, nerve microtissues and PRP in reconstructing a 12-mm tibial nerve defect and to explore the value of multimodal ultrasound techniques in evaluating the prognosis of nerve repair. METHODS: In vitro, nerve microtissue activity was first investigated, and the effects on SC proliferation, migration, factor secretion, and axonal regeneration of dorsal root ganglia (DRG) were evaluated by coculture with nerve microtissues and PRP. In vivo, seventy-five rabbits were equally and randomly divided into Hollow, PRP, Micro-T (Microtissues), Micro-T + PRP and Autograft groups. By analysing the neurological function, electrophysiological recovery, and the comparative results of multimodal ultrasound and histological evaluation, we investigated the effect of these new nerve grafts in repairing tibial nerve defects. RESULTS: Our results showed that the combined application of nerve microtissues and PRP could significantly promote the proliferation, secretion and migration of SCs and the regeneration of axons in the early stage. The Micro-T + PRP group and Autograft groups exhibited the best nerve repair 12 weeks postoperatively. In addition, the changes in target tissue stiffness and microvascular perfusion on multimodal ultrasound (shear wave elastography; contrast-enhanced ultrasonography; Angio PlaneWave UltrasenSitive, AngioPLUS) were significantly correlated with the histological results, such as collagen area percentage and VEGF expression, respectively. CONCLUSION: Our novel tissue-engineered nerve graft shows excellent efficacy in repairing 12-mm defects of the tibial nerve in rabbits. Moreover, multimodal ultrasound may provide a clinical reference for prognosis by quantitatively evaluating the stiffness and microvescular flow of nerve grafts and targeted muscles.

Hepatitis B virus in cerebrospinal fluid of a patient with purulent bacterial meningitis detected by multiplex-PCR: A case report.

BACKGROUND: Bacterial meningitis (BM) is a common central nervous system inflammatory disease. BM may cause serious complications, and early diagnosis is essential to improve the prognosis of affected patients. CASE SUMMARY: A 37-year-old man was hospitalized with purulent meningitis because of worsening headache for 12 h, accompanied by vomiting, fever, and rhinorrhea. Head computed tomography showed a lesion in the left frontal lobe. Infectious disease screening showed positivity for hepatitis B surface antigen, hepatitis B e antigen, and hepatitis B core antigen. Cerebrospinal fluid (CSF) leak was suspected based on clinical history. Streptococcus pneumoniae (S. pneumoniae) was detected in CSF by metagenomic next-generation sequencing (mNGS) technology, confirming the diagnosis of purulent BM. After treatment, multiplex PCR indicated the presence of hepatitis B virus (HBV) DNA and absence of S. pneumoniae DNA in CSF samples. CONCLUSION: We report a rare case of HBV in the CSF of a patient with purulent BM. Multiplex PCR is more sensitive than mNGS for detecting HBV DNA.

Bilayered scaffold with 3D printed stiff subchondral bony compartment to provide constant mechanical support for long-term cartilage regeneration.

BACKGROUND/OBJECTIVE: We seek to figure out the effect of stable and powerful mechanical microenvironment provided by Ti alloy as a part of subchondral bone scaffold on long-term cartilage regeneration.Methods: we developed a bilayered osteochondral scaffold based on the assumption that a stiff subchondral bony compartment would provide stable mechanical support for cartilage regeneration and enhance subchondral bone regeneration. The subchondral bony compartment was prepared from 3D printed Ti alloy, and the cartilage compartment was created from a freeze-dried collagen sponge, which was reinforced by poly-lactic-co-glycolic acid (PLGA). RESULTS: In vitro evaluations confirmed the biocompatibility of the scaffold materials, while in vivo evaluations demonstrated that the mechanical support provided by 3D printed Ti alloy layer plays an important role in the long-term regeneration of cartilage by accelerating osteochondral formation and its integration with the adjacent host tissue in osteochondral defect model at rabbit femoral trochlea after 24 weeks. CONCLUSION: Mechanical support provided by 3D printing Ti alloy promotes cartilage regeneration by promoting subchondral bone regeneration and providing mechanical support platform for cartilage synergistically. TRANSLATIONAL POTENTIAL STATEMENT: The raw materials used in our double-layer osteochondral scaffolds are all FDA approved materials for clinical use. 3D printed titanium alloy scaffolds can promote bone regeneration and provide mechanical support for cartilage regeneration, which is very suitable for clinical scenes of osteochondral defects. In fact, we are conducting clinical trials based on our scaffolds. We believe that in the near future, the scaffold we designed and developed can be formally applied in clinical practice.

Applying the pathogen-targeted next-generation sequencing method to pathogen identification in cerebrospinal fluid.

BACKGROUND: The cerebrospinal fluid (CSF) culture is a widely used method for the diagnosis of meningitis, but its detection sensitivity is low. Several new methods have been developed for pathogen detection, including metagenomic next-generation sequencing (mNGS) and pathogen-targeted NGS (ptNGS). In this study, we aimed to evaluate the performance of ptNGS in pathogen detection in CSF. METHODS: CSF specimens were acquired from 38 patients with meningitis who were diagnosed at Xuanwu Hospital, Capital Medical University between October 2020 and February 2021. DNA was extracted from the CSF samples, and pathogens were identified using both ptNGS and mNGS. SPSS 22.0 software was used to compare the pathogen detection performance of ptNGS and mNGS in CSF. RESULTS: Among the 38 patients with meningitis, 14 had a non-infectious disease (NID) and 24 had an infectious disease (ID). Of the 38 samples, both ptNGS and mNGS detected 9 (23.7%) positive samples, and 12 (31.6%) negative samples. Thirteen (34.2%) samples were detected to be positive by ptNGS only, and 4 (10.5%) were detected to be positive by mNGS only. The positivity rate detected by ptNGS for the ID group was higher than that detected by mNGS (P=0.080), and the positivity rates detected by ptNGS and mNGS for the NID group were comparable. The positive predictive value (PPV) and negative predictive value (NPV) of diagnosing an ID by ptNGS were 77.3% and 56.3%, respectively. While, the PPV and NPV of diagnosing an ID by mNGS were 76.9% and 44.0%, respectively. ptNGS increased the sensitivity rate by approximately 70%. The sensitivity rate of ptNGS was higher than that of mNGS (70.8% vs. 41.7%), while the specificity rate of mNGS was higher than that of ptNGS (78.6% vs. 64.3%). Additionally, ptNGS required a shorter time for pathogen diagnosis (15 vs. 24 hrs) and had lower costs than mNGS. CONCLUSIONS: ptNGS has a number of advantages over mNGS, including its sensitivity, timeliness, and economy, all factors that are important considerations in clinical use.

A novel tissue engineered nerve graft constructed with autologous vein and nerve microtissue repairs a long-segment sciatic nerve defect.

Veins are easy to obtain, have low immunogenicity, and induce a relatively weak inflammatory response. Therefore, veins have the potential to be used as conduits for nerve regeneration. However, because of the presence of venous valves and the great elasticity of the venous wall, the vein is not conducive to nerve regeneration. In this study, a novel tissue engineered nerve graft was constructed by combining normal dissected nerve microtissue with an autologous vein graft for repairing 10-mm peripheral nerve defects in rats. Compared with rats given the vein graft alone, rats given the tissue engineered nerve graft had an improved sciatic static index, and a higher amplitude and shorter latency of compound muscle action potentials. Furthermore, rats implanted with the microtissue graft had a higher density and thickness of myelinated nerve fibers and reduced gastrocnemius muscle atrophy compared with rats implanted with the vein alone. However, the tissue engineered nerve graft had a lower ability to repair the defect than autogenous nerve transplantation. In summary, although the tissue engineered nerve graft constructed with autologous vein and nerve microtissue is not as effective as autologous nerve transplantation for repairing long-segment sciatic nerve defects, it may nonetheless have therapeutic potential for the clinical repair of long sciatic nerve defects. This study was approved by the Experimental Animal Ethics Committee of Chinese PLA General Hospital (approval No. 2016-x9-07) on September 7, 2016.

Ultrasound-guided platelet-rich plasma injection and multimodality ultrasound examination of peripheral nerve crush injury.

Ultrasound-guided platelet-rich plasma (PRP) injection is able to make up for the limitations of applying a single growth factor. The goal of this study was to investigate the effects of serial ultrasound-guided PRP injections of the appropriate concentration on the treatment of sciatic nerve crush injury, and explore the value of multimodality ultrasound techniques in evaluating the prognosis of crushed peripheral nerve. In vitro, optimal concentration of PRP (from 150%, 250%, 450%, and 650%) was screened due for its maximal effect on proliferation and neurotrophic function of Schwann cells (SCs). In vivo, ninety rabbits were equally and randomly divided into normal control, model, PRP-2.5×, PRP-4.5×, and PRP-6.5× groups. The neurological function and electrophysiological recovery evaluation, and the comparison of the multimodality ultrasound evaluation with the histological results of sciatic nerve crush injury were performed to investigate the regenerative effects of PRP at different concentrations on the sciatic nerve crush injury. Our results showed that the PRP with a 4.5-fold concentration of whole blood platelets could significantly stimulate the proliferation and secretion of SCs and nerve repair. The changes in stiffness and blood perfusion were positively correlated with the collagen area percentage and VEGF expression in the injured nerve, respectively. Thus, serial ultrasound-guided PRP injections at an appropriate concentration accelerates the recovery of axonal function. Multimodality ultrasound techniques provide a clinical reference for prognosis by allowing the stiffness and microcirculation perfusion of crush-injured peripheral nerves to be quantitatively evaluated.

An automatic method for counting wheat tiller number in the field with terrestrial LiDAR.

BACKGROUND: The tiller number per unit area is one of the main agronomic components in determining yield. A real-time assessment of this trait could contribute to monitoring the growth of wheat populations or as a primary phenotyping indicator for the screening of cultivars for crop breeding. However, determining tiller number has been conventionally dependent on tedious and labor-intensive manual counting. In this study, an automatic tiller-counting algorithm was developed to estimate the tiller density under field conditions based on terrestrial laser scanning (TLS) data. The novel algorithm, which is named ALHC, involves two steps: (1) the use of an adaptive layering (AL) algorithm for cluster segmentation and (2) the use of a hierarchical clustering (HC) algorithm for tiller detection among the clusters. Three field trials during the 2016-2018 wheat seasons were conducted to validate the algorithm with twenty different wheat cultivars, three nitrogen levels, and two planting densities at two ecological sites (Rugao & Xuzhou) in Jiangsu Province, China. RESULT: The results demonstrated that the algorithm was promising across different cultivars, years, growth stages, planting densities, and ecological sites. The tests from Rugao and Xuzhou in 2016-2017 and Rugao in 2017-2018 showed that the algorithm estimated the tiller number of the wheat with regression coefficient (R2) values of 0.61, 0.56 and 0.65, respectively. In short, tiller counting with the ALHC generally underestimated the tiller number and performed better for the data with lower plant densities, compact plant types and the jointing stage, which were associated with overlap and noise between plants and inside the dense canopy. CONCLUSIONS: Differing from the previous methods, the ALHC proposed in this paper made full use of 3D crop information and developed an automatic tiller counting method that is suitable for the field environment.

The effect of a diet based on rice straw co-fermented with probiotics and enzymes versus a fresh corn Stover-based diet on the rumen bacterial community and metabolites of beef cattle.

Improvement of the food value of rice straw is urgently required in rice crop growing areas to mitigate pollution caused by rice straw burning and enhance the supply of high-quality forages for ruminants. The aims of the present study were to compare the effects of fresh corn Stover and rice straw co-fermented with probiotics and enzymes on rumen fermentation and establish the feasibility of increasing the rice straw content in ruminant diets and, by extension, reducing air pollution caused by burning rice straw. Twenty Simmental hybrid beef cattle were randomly allotted to two groups with ten cattle per group. They were fed diets based either on rice straw co-fermented with probiotics and enzymes or fresh corn Stover for 90 days. Rumen fluid was sampled with an esophageal tube vacuum pump device from each animal on the mornings of days 30, 60, and 90. Bacterial diversity was evaluated by sequencing the V4-V5 region of the 16S rRNA gene. Metabolomes were analyzed by gas chromatography/time-of-flight mass spectrometry (GC-TOF/MS). Compared to cattle fed fresh corn Stover, those fed rice straw co-fermented with probiotics and enzymes had higher (P < 0.05) levels of acetic acid and propionate in rumen liquid at d 60 and d 90 respectively, higher (P < 0.05) abundances of the phyla Bacteroidetes and Fibrobacteres and the genera Ruminococcus, Saccharofermentans, Pseudobutyrivibrio, Treponema, Lachnoclostridium, and Ruminobacter, and higher (P < 0.05) concentrations of metabolites involved in metabolisms of amino acid, carbohydrate, and cofactors and vitamins. Relative to fresh corn Stover, rice straw co-fermented with probiotics and enzymes resulted in higher VFA concentrations, numbers of complex carbohydrate-decomposing and H2-utilizing bacteria, and feed energy conversion efficiency in the rumen.

Platelet-Rich Plasma Therapy in the Treatment of Diseases Associated with Orthopedic Injuries.

Platelet-rich plasma (PRP) is an autologous platelet concentrate prepared from the whole blood that is activated to release growth factors (GFs) and cytokines and has been shown to have the potential capacity to reduce inflammation and improve tissue anabolism for regeneration. The use of PRP provides a potential for repair due to its abundant GFs and cytokines, which are key in initiating and modulating regenerative microenvironments for soft and hard tissues. Among outpatients, orthopedic injuries are common and include bone defects, ligament injury, enthesopathy, musculoskeletal injury, peripheral nerve injury, chronic nonhealing wounds, articular cartilage lesions, and osteoarthritis, which are caused by trauma, sport-related or other types of trauma, or tumor resection. Surgical intervention is often required to treat these injuries. However, for numerous reasons regarding limited regeneration capacity and insufficient blood supply of the defect region, these treatments commonly result in unsatisfactory outcomes, and follow-up treatment is challenging. The aim of the present review is to explore future research in the field of PRP therapy in the treatment of diseases associated with orthopedic injuries. Impact statement In recent years, platelet-rich plasma (PRP) has become widely used in the treatment of diseases associated with orthopedic injuries, and the results of numerous studies are encouraging. Due to diseases associated with orthopedic injuries being common in clinics, as a conservative treatment, more and more doctors and patients are more likely to accept PRP. Importantly, PRP is a biological product of autologous blood that is obtained by a centrifugation procedure to enrich platelets from whole blood, resulting in few complications, such as negligible immunogenicity from an autologous source, and it is also simple to produce through an efficient and cost-effective method in a sterile environment. However, the applicability, advantages, and disadvantages of PRP therapy have not yet been fully elucidated. The aim of the present review is to explore future research in the field of PRP therapy in the treatment of diseases associated with orthopedic injuries, as well as to provide references for clinics.

Administration of All-Trans Retinoic Acid to Pregnant Sows Improves the Developmental Defects of Hoxa1-/- Fetal Pigs.

Hoxa1 mutation adversely affect fetal pig development, but whether all-trans retinoic acid (ATRA) administration to Hoxa1+/- pregnant sows can improve Hoxa1-/- fetal pig development defects has not been reported. A total of 24 healthy Hoxa1+/- sows were mated with a healthy Hoxa1+/- boar and randomly assigned to one control group and nine experiment groups. ATRA was orally administered to pregnant sows at the doses of 0, 4, 5, or 6 mg/kg maternal body weight on 12, 13, and 14 days post coitum (dpc), respectively, and a total of 146 live piglets were delivered including 37 Hoxa1-/- piglets and 109 non-Hoxa1-/- piglets. Results indicated that Hoxa1-/- piglets delivered by sows in control group had bilateral microtia, canal atresia and ear's internal defects, and had lower birth liveweight and external ear score than non-Hoxa1-/- neonatal piglets (P < 0.05). Maternal administration with ATRA can effectively correct the development defects of Hoxa1-/- fetal pigs, Hoxa1-/- neonatal piglets delivered by sows administered ATRA at a dose of 4 mg/kg body weight on 14 dpc had higher birth liveweight (P > 0.05) and higher scores of external ear (P < 0.05) compared to Hoxa1-/- neonatal piglets from the control group, but had no significantly difference in terms of birth liveweight and external ear integrity than non-Hoxa1-/- piglets from the control group (P > 0.05). The time of ATRA administration significantly affected Hoxa1-/- fetal development (P < 0.05). Administration of ATRA to Hoxa1+/- pregnant sows at 4 mg/kg body weight on 14 dpc can effectively improve the birth liveweight and ear defects of Hoxa1-/- piglets.

MicroRNA-181 inhibits glioblastoma cell growth by directly targeting CCL8.

MicroRNAs (miRNAs/miRs), including miR-181, are closely linked to the development and progression of glioblastoma. However, the function of miR-181 in glioblastoma has not been fully clarified. The aim of the present study was to investigate the role of miR-181 in glioblastoma. miR-181 was revealed to be downregulated in glioblastoma tissues and cell lines, and associated with poor prognosis in patients with glioblastoma. Overexpression of miR-181 inhibited glioblastoma cell proliferation, invasion and migration, arrested glioblastoma cell cycle in the G1 phase and induced glioblastoma cell apoptosis. miR-181 was demonstrated to decrease expression of C-C motif chemokine ligand 8 (CCL8) by directly interacting with its 3'-untranslated region. Overexpression of CCL8 inversely reversed the proliferation, invasion and migration-promoting effects of miR-181 in glioblastoma cells. Furthermore, CCL8 was upregulated in glioblastoma tissues and was negatively correlated with miR-181 expression. These results indicate that miR-181 is a potential molecular biomarker or therapeutic target in the clinical management of glioblastoma.

Limbic Encephalitis Associated with Anti-γ-aminobutyric Acid B Receptor Antibodies: A Case Series from China.

BACKGROUND: Autoimmune encephalitis associated with antibodies against γ-aminobutyric acid B receptor (GABA B R) in patients with limbic encephalitis (LE) was first described in 2010. We present a series of Han Chinese patients for further clinical refinement. METHODS: Serum and cerebrospinal fluid (CSF) samples from patients referred to the program of encephalitis and paraneoplastic syndrome of Peking Union Medical College Hospital were tested with indirect immunofluorescence. Clinical information of patients with anti-GABA B R antibody positivity was retrospectively reviewed, and descriptive statistical analysis was performed. RESULTS: All eighteen anti-GABA B R antibody-positive cases had limbic syndromes, and electroencephalogram (EEG) or neuroimaging evidence fulfilled the diagnostic criteria of LE. Four patients had additional antibodies against Hu in serum and one had anti-N-methyl-d-aspartate receptor antibody in both sera and CSF. Seventeen (17/18) patients presented with new-onset refractory seizure or status epileptics. Twelve (12/18) patients had memory deficits, 11 (11/18) patients had personality change, 7 (7/18) patients had disturbance of consciousness, and 3 (3/18) patients showed cerebellar dysfunction. One patient with LE had progressive motor and sensory polyneuropathy. Lung cancer was detected in 6 (6/18) patients. Ten (10/18) patients showed abnormality in bilateral or unilateral mediotemporal region on magnetic resonance imaging. Ten (10/18) patients had temporal lobe epileptic activity with or without general slowing on EEG. Seventeen patients received immunotherapy and 15 of them showed neurological improvement. Four patients with lung cancer died within 1-12 months due to neoplastic complications. CONCLUSIONS: Our study demonstrates that most Han Chinese patients with anti-GABA B R antibody-associated LE have prominent refractory epilepsy and show neurological improvement on immunotherapy. Patients with underlying lung tumor have a relatively poor prognosis. Testing for anti-GABA B R antibodies is necessary for patients with possible LE or new-onset epilepsy with unknown etiology.

Comprehensive detection and identification of seven animal coronaviruses and human respiratory coronavirus 229E with a microarray hybridization assay.

Based on microarray hybridization, a diagnostic test for coronavirus infection was developed using eight coronavirus strains: canine coronavirus (CCoV), feline infectious peritonitis virus (FIPV), feline coronavirus (FCoV), bovine coronavirus (BCoV), porcine respiratory coronavirus (PRCoV), turkey enteritis coronavirus (TCoV), transmissible gastroenteritis virus (TGEV), and human respiratory coronavirus (HRCoV). Up to 104 cDNA clones of eight viruses were obtained by reverse transcription PCR with different pairs of primers designed for each virus and a pair of universal primers designed for the RNA polymerase gene of coronavirus. Total RNAs extracted from virus were reverse transcribed, followed by multi-PCR amplification and labeled with Cy3-dCTP. All labeled cDNAs and prepared gene chips were subjected to specific hybridization. The results showed that extensive cross-reaction existed between CCoV, FCoV, FIPV, TGEV and PRCoV, while there was no cross-reaction between BCoV, TCoV and HRCoV. The ultimate specific gene chip was developed with DNA fragments reamplified from the chosen recombinant plasmids without cross-reaction between different coronaviruses. The hybridization results showed that this gene chip could specifically identify and distinguish the eight coronaviruses and the sensitivity of the chip may be 1,000x more sensitive than PCR, indicating that it can be used for the diagnosis of eight coronavirus infections at the same time.