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Paroxysmal kinesigenic dyskinesias is a paroxysmal movement disorder characterized by recurrent, brief attacks of abnormal involuntary movements induced by sudden voluntary movements. Although several loci, including the pericentromeric region of chromosome 16, have been linked to paroxysmal kinesigenic dyskinesias, the causative gene has not yet been identified. Here, we identified proline-rich transmembrane protein 2 (PRRT2) as a causative gene of paroxysmal kinesigenic dyskinesias by using a combination of exome sequencing and linkage analysis. Genetic linkage mapping with 11 markers that encompassed the pericentromeric of chromosome 16 was performed in 27 members of two families with autosomal dominant paroxysmal kinesigenic dyskinesias. Then, the whole-exome sequencing was performed in three patients from these two families. By combining the defined linkage region (16p12.1-q12.1) and the results of exome sequencing, we identified an insertion mutation c.649_650InsC (p.P217fsX7) in one family and a nonsense mutation c.487C>T (p.Q163X) in another family. To confirm our findings, we sequenced the exons and flanking introns of PRRT2 in another three families with paroxysmal kinesigenic dyskinesias. The c.649_650InsC (p.P217fsX7) mutation was identified in two of these families, whereas a missense mutation, c.796C>T (R266W), was identified in another family with paroxysmal kinesigenic dyskinesias. All of these mutations completely co-segregated with the phenotype in each family. None of these mutations was identified in 500 normal unaffected individuals of matched geographical ancestry. Thus, we have identified PRRT2 as the first causative gene of paroxysmal kinesigenic dyskinesias, warranting further investigations to understand the pathogenesis of this disorder. [ABSTRACT FROM AUTHOR]

With consideration of motion control performance and efficient information communication, the synchronization problem on communication connectivity preservation and guaranteed consensus performance for networked mechanical systems has attracted considerable attention in recent years. Different from the existing works, this article investigates a brand-new appointed-time consensus control approach for uncertain networked Euler-Lagrange systems on a directed graph via exploring the prescribed performance control structure. First, a two-layer prescribed performance envelope is formulated via using an appointed-time convergent function for position-related and velocity-related consensus errors, respectively. Then, a simple state-feedback virtual controller with online adaptive performance adjustment is developed to preserve the communication connectivity. Moreover, to guarantee the velocity consensus of the networked systems and improve the position consensus accuracy, an appointed-time adaptive controller is designed by applying the norm inequality to the system uncertainties and external disturbances. Compared to the existing consensus control approaches, the prime advantage of the proposed one is that the constraints generated from the communication ranges are approximated by a time-varying contractive performance envelope, wherein, the appointed-time convergence and steady-state tracking accuracy are preassigned a priori. Meanwhile, no repeated logarithmic error transformations are required in the relevant controller design, which implies that the complexity of the devised control laws has decreased dramatically. Finally, two groups of illustrative examples are organized to validate the effectiveness of the proposed consensus control approach.

Four undescribed flavonoid glucosides (iridins B-C, tectoridin A and ampelopsinin A); one undescribed phenolic glucoside (diplostephioside B); one undescribed phenolic compound (phenanthrenetriol A); and seventeen known compounds were isolated from the rhizomes of Iris domestica. The chemical structures of the undescribed compounds were established by spectroscopic/spectrometric data interpretation using HRESIMS, NMR, and ECD. Tectoridin A, nigricin A and naringenin exhibited anti-inflammatory activities with inhibition rates of 53.71%, 57.68% and 88.71%, respectively, against the NF-κB signaling pathway at a concentration of 10 μM. 4'-O-methylnyasol (10 μM) exhibited 84.91% antiproliferative activity against the K562 human leukemia cell line with an IC 50 value of 4.20 μM., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

A pair of stilbenes with γ-lactam unit [(+)- 1 and (-)- 1 ], a new phenolic glucoside ( 2 ), and a new isoflavone glucoside ( 3 ), together with two known compounds ( 4-5 ) were isolated from the rhizomes of Belamcanda chinensis . The chemical structures of the undescribed compounds were elucidated on the basis of detailed spectroscopic analyses. Compounds 1 , 4 , and 5 (10 μM) exhibited anti-inflammatory activities with inhibition rates of 30.46%, 60.34%, and 37.91%, respectively, against the NF-κB signaling pathway.

Histone methylation, particularly at the H3K4 position, is thought to contribute to the specification of photoreceptor cell fate; however, the mechanisms linking histone methylation with transcription factor transactivation and photoreceptor gene expression have not yet been determined. Here, we demonstrate that MLL5 is abundantly expressed in the mouse retina. Mll5 deficiency impaired electroretinogram responses, alongside attenuated expression of a number of retina genes. Mechanistic studies revealed that MLL5 interacts with the retina-specific transcription factor, CRX, contributing to its binding to photoreceptor-specific gene promoters. Moreover, depletion of MLL5 impairs H3K4 methylation and H3K79 methylation, which subsequently compromises CRX-CBP assembly and H3 acetylation on photoreceptor promoters. Our data support a scenario in which recognition of H3K4 methylation by MLL5 is required for photoreceptor-specific gene transcription through maintaining a permissive chromatin state and proper CRX-CBP recruitment at promoter sites., Competing Interests: The authors declare no competing interests., (© 2022 The Author(s).)

The zebrafish as an alternative animal model for developmental toxicity testing has been extensively investigated, but its assay protocol was not harmonized yet. This study has validated and optimized the zebrafish developmental toxicity assay previously reported by multiple inter-laboratory studies in the United States and Europe. In this study, using this classical protocol, of 31 ICH-positive compounds, 23 compounds (74.2%) were teratogenic in zebrafish, five had false-negative results, and three were neither teratogenic nor non-teratogenic according to the protocol standard; of 14 ICH-negative compounds, 12 compounds (85.7%) were non-teratogenic in zebrafish and two had false-positive results. After we added an additional TI value in the zebrafish treated with testing compounds at 2 dpf along with the original 5 dpf, proposed a new category as the uncategorized compounds for those TI values smaller than the cutoff both at 2 dpf and 5 dpf but inducing toxic phenotypes, refined the testing concentration ranges, and optimized the TI cut-off value from ≥ 10 to ≥ 3 for compounds with refined testing concentrations, this optimized zebrafish developmental assay reached 90.3% sensitivity (28/31 positive compounds were teratogenic in zebrafish) and 88.9% (40/45) overall predictability. Our results from this study strongly support the use of zebrafish as an alternative in vivo method for screening and assessing the teratogenicity of candidate drugs for regulatory acceptance., Competing Interests: M-ZD, Y-FH, YP, YZ, and C-QL are employed by Hunter Biotechnology, Inc., which is mainly specialized in developing and marketing novel and innovative in vivo zebrafish-based assays for the following industries worldwide: Pharmaceutical, Biotechnology, Environmental, Natural Products, Industrial Chemical Products, Agrochemical, Food Additives, Skin Beauty Products, and Nutraceuticals. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Song, Dai, Zhu, Huang, Liu, Zhang, Xie, Peng, Zhang, Li and Zhang.)

Background: Human papillomavirus (HPV) type 16 accounts for a larger share of cervical cancer and has been a major health problem worldwide for decades. The progression of initial infection to cervical cancer has been linked to viral sequence properties; however, the role of HPV16 variants in the risk of cervical carcinogenesis, especially with longitudinal follow-up, is not fully understood in China., Methods: We aimed to investigate the genetic variability of HPV16 E6 and E7 oncogenes in isolates from cervical exfoliated cells. Between December 2012 and December 2014, a total of 310 single HPV16-positive samples were selected from women living in the Taizhou area, China. Sequences of all E6 and E7 oncogenes were analysed by PCR-sequencing assay. Detailed sequence comparison, genetic heterogeneity analyses and maximum-likelihood phylogenetic tree construction were performed with BioEdit Sequence Alignment Editor and MEGA X software. Data for cytology tests and histological diagnoses were obtained from our Taizhou Area Study with longitudinal follow-up for at least 5 years. The relationship between HPV16 variants and cervical carcinogenesis risk was analysed by the chi-square test or Fisher's exact test., Results: In this study, we obtained 64 distinct variation patterns with the accession GenBank numbers MT681266-MT681329. Phylogenetic analysis revealed that 98.3% of HPV16 variants belong to lineage A, in which the A4 (Asian) sublineage was dominant (64.8%), followed by A2 (12.1%), A1 (11.4%), and A3 (10.0%). The A4 (Asian) sublineage had a higher risk of CIN2+ than the A1-3 (European) sublineages (OR = 2.69, 95% CI = 1.04-6.97, P < 0.05). Furthermore, nucleotide variation in HPV16 E6 T178G is associated with the development of cervical cancer., Conclusion: These data could provide novel insights into the role of HPV16 variants in cervical carcinogenesis risk in China.

Conventional mammalian ischemic stroke models for drug screening are technically challenging, laborious and time-consuming. In this study, using Ponatinib as an inducer, we developed and characterized a zebrafish ischemic stroke model. This zebrafish ischemic stroke had the cerebral vascular endothelial injury, thrombosis, reduced blood flow, inflammation and apoptosis as well as the reduced motility. The zebrafish ischemic stroke model was validated with 6 known human therapeutic drugs of ischemic stroke (Aspirin, Clopidogrel, Naoxintong capsules, Edaravone, Xingnaojing injection, Shuxuening injection). The mRNA levels of the neovascularization-related gene (vegfaa) and vascular endothelial growth factor receptor gene (VEGFR), neurodevelopment related genes (mbp and α1-tubulin), brain-derived neurotrophic factor (BDNF) and glial cell derived neurotrophic factor (GDNF) were significantly downregulated; whereas apoptosis-related genes (caspase-3, caspase-7, caspase-9 and bax/bcl-2), and inflammatory factor genes (IL-1β, IL-6, IL-10, TNF-α and NF-κB) were remarkably upregulated in the model. These results suggest that the pathophysiology of Ponatinib-induced zebrafish ischemic stroke is similar to that of human ischemic stroke patients and this whole animal model could be used to study the complex cellular and molecular pathogenesis of ischemic stroke and to rapidly identify therapeutic agents., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Aims: Current treatment options for ovarian clear cell carcinoma (OCCC) are limited to combination of platinum-based and other cytotoxic agents to which patients respond poorly due to intrinsic chemoresistance. There is therefore an urgent need to develop alternative therapeutic strategies for OCCC. Results: Cysteine deprivation suppresses OCCC growth in vitro and in vivo with no apparent toxicity. Modes of cell death induced by cysteine deprivation in OCCC are determined by their innate metabolic profiles. Cysteine-deprived glycolytic OCCC is abolished primarily by oxidative stress-dependent necrosis and ferroptosis, which can otherwise be prevented by pretreatment with antioxidative agents. Meanwhile, OCCC that relies on mitochondria respiration for its bioenergetics is suppressed through apoptosis, which can otherwise be averted by pretreatment with cysteine precursor alone, but not with antioxidative agents. Cysteine deprivation induces apoptosis in respiring OCCC by limiting iron-sulfur (Fe-S) cluster synthesis in the mitochondria, without which electron transport chain may be disrupted. Respiring OCCC responds to Fe-S cluster deficit by increasing iron influx into the mitochondria, which leads to iron overload, mitochondria damage, and eventual cell death. Innovation/Conclusion: This study demonstrates the importance of cysteine availability in OCCC that is for its antioxidative property and its less appreciated role in mitochondria respiration. Regardless of OCCC metabolic profiles, cysteine deprivation abolishes both glycolytic and respiring OCCC growth in vitro and in vivo . Conclusion: This study highlights the therapeutic potential of cysteine deprivation for OCCC.

This study was aimed to assess effects of three strains of probiotics Lactobacillus acidophilus NCFM, Lactobacillus rhamnosus HN001, and Bifidobacterium animalis subsp . lactis Bi-07 on the intestinal motility and inflammation in the zebrafish models. The intestinal motility model was established using 5 days postfertilization (dpf) zebrafish administered with a fluorescent dye Nile red at 10 ng/mL for 16 h, followed by probiotics treatment for 24 h and the intestinal motility was inversely proportional to the intestinal fluorescence intensity that was quantitatively measured by image analysis. The intestinal inflammation was induced by treating 3 dpf neutrophil fluorescent zebrafish with 0.0125% of trinitrobenzenesulfonic acid for 48 h. Probiotics were administered at low, moderate, and high concentrations determined based on maximum tolerable concentration through soaking. All three strains of probiotics promoted intestinal movement, of which B. animalis subsp. lactis Bi-07 was most potent at lower concentrations. L. rhamnosus HN001 and B. animalis subsp. lactis Bi-07 had the therapeutic effects on the intestinal inflammation and the inflammation-associated mucosal damage recovery. The anti-inflammatory mechanisms of L. rhamnosus HN001 was related to both reduce inflammatory factor interleukin-6 ( IL-6 ) and restored tissue repair factor transforming growth factor-β-1 ( TGFβ-1 ); whereas B. animalis subsp. lactis Bi-07 was probably only associated with TGFβ-1 elevation. Using larval zebrafish models for probiotics screening and assessment would speed up product research and development and improve products' efficacy and quality.

Background: Non-invasive prenatal testing (NIPT) has been established as a routine prenatal screening to assess the risk of common foetal aneuploidy disorder (trisomy 21, 18, and 13). NIPT has high sensitivity and high specificity, but false positive and false negative results still exist. False negative NIPT results involving Down syndrome are rare, but have a high clinical impact on families and society., Case Presentation: We described a case of a foetus that tested "negative" for trisomy 21 (Z-score was 0.664) by NIPT based on the semiconductor sequencing platform (SSP). The foetal fraction of cell-free DNA was 16.9%; this percentage was much larger than the threshold of 4% for obtaining accurate NIPT results. However, postnatally, the newborn was diagnosed with Down syndrome with the 46,XY,der(21;21)(q10;q10),+ 21 karyotype., Conclusions: We presented a case of false negative NIPT results, which may occur through biological mechanisms rather than poor quality, technical errors or negligence. It is imperative for clinical geneticists and their patients to understand that NIPT is still a screening test.

Fenobucarb (2-sec-butylphenyl methylcarbamate, BPMC) is an extensively used carbamate insecticide. Its developmental neurotoxicity and the underlying mechanisms have not been well investigated. In this study, zebrafish embryos were exposed to various concentrations of BPMC from 6 hpf (hours post fertilization, hpf) to 120 hpf. BPMC induced developmental toxicity with reduced motility in larval zebrafish. The spinal cord neutrophil infiltration, increased ROS production, caspase 3 and 9 activation, central nerve and peripheral motor neuron damage, axon and myelin degeneration were observed in zebrafish treated with BPMC generally in a dose-dependent manner. The expression of eight marker genes for nervous system function or development, namely, a1-tubulin, shha, elavl3, gap43, syn2a, gfap, mbp and manf, was significantly downregulated following BPMC exposure. AChE activity reduction and ache gene expression suppression was also found significantly in BPMC-treated zebrafish. These results indicate that BPMC is highly toxic to zebrafish and that BPMC induces zebrafish developmental neurotoxicity through pathways involved in inflammation, oxidative stress, degeneration and apoptosis., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2020. Published by Elsevier B.V.)

Introduction: This study was aimed to assess uric acid (UA)-lowering effect and its possible mechanisms of a natural complex product Yaocha in a live zebrafish model., Methods: The zebrafish high UA model was established by feeding 5 dpf zebrafish with both an uricase inhibitor potassium oxonate at 10 mM and an UA synthesis precursor xanthine sodium at 0.5 mM for 24 h. Yaocha was administered to the high UA zebrafish through soaking at 3 various concentrations, with allopurinol as a positive control. UA level, xanthine oxidase (XOD) activity, and mRNA expression of hypoxanthine guanine-phosphoribosyltransferases transferase (HPRT1) and organic anion transporter 1 (OAT1) were measured., Results: Yaocha effectively reduced UA level and inhibited xanthine oxidase (XO) activity in the high UA zebrafish. Yaocha could be a potential therapeutics for hyperuricemia through up-regulating HPRT1 and OAT1 gene expression and suppressing XO activity., Discussion: These results suggested that Yaocha hold a potential for high UA prevention and therapy, possibly through inhibiting UA production and promoting urate secretion and purine conversion., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Objective: Yun Kang oral liquid is a listed proprietary Chinese Medicine. To further evaluate its efficacy, this experiment established a kidney deficiency and luteum inhibition threatened abortion rat model to observe the effects of Yun Kang oral liquid., Methods: Sixty pregnant rats were randomly divided into normal control group (NC), model group (MG), dydrogesterone group (DT, 3.02 mg/kg), and Yun Kang oral liquid low-dose group (YK-L, 4 ml/kg), medium dose group (YK-M, 6 ml/kg), high dose group (YK-H, 9 ml/kg), 10 in each group. On the first day of pregnancy, each administration group was treated with the test drug at the prescribed dose every morning, and the NC group and the MG group were given an equal volume of purified water for 10 days; the rats were intragastrically administrated every afternoon, except for the NC group. In addition, the other groups were intragastrically administered with hydroxyurea at a dose of 450 mg/kg for 9 days, and mifepristone was administered at a dose of 4.0 mg/kg on the 10th day. On the 9th day of pregnancy, behavioral signs such as back temperature, grasping force, pain threshold, and autonomic activity were measured in each group. On the 11th day of pregnancy, blood was collected from the abdominal aorta in each group to determine serum levels of estradiol (E 2 ) , progesterone (P) and thromboxane B 2 (TXB 2 ) . Ovary and fetal uterus were removed, the number and diameter of embryos were observed, and the ovary and uterus indexes were calculated., Results: Compared with the NC group, the back temperature, grip, pain threshold, number of spontaneous activities, number of embryos, embryo diameter, uterus index and serum E 2 , P, TXB 2 levels in the MG group were decreased significantly (P＜0.05, 0.01). Compared with the MG group, the back temperature, grasping force, number of embryos, embryo diameter and serum E 2 and P levels were increased significantly in each dose group (P＜0.05, 0.01); the pain threshold, autonomic activity, and uterus index of YK-M and YK-H group were increased significantly (P＜0.05); serum level of TXB 2 in YK-H group were increased significantly (P＜0.05)., Conclusion: Yun Kang oral liquid has a clear kidney-filling effect on rats with threatened abortion caused by kidney deficiency-luteal suppression. The mechanism may be related to raising serum E 2 , P, TXB 2 levels, improving kidney deficiency and improving embryo quality.

Objective: Through the establishment of abortion model caused by embryo implantation difficulties, exploring the role of Yun Kang oral liquid in protecting embryos., Methods: The pregnant rats were divided into 6 groups:normal control group (NC), model group (MG), dydrogesterone group (DT), and three dose groups of low, medium and high levels of Yun Kang oral liquid (YK-L, YK-M, YK-H), 11 in each group.From the first day of pregnancy, daily intragastric administration, the dose of DT group was 3.02 mg/kg, and the doses of Yun Kang oral liquid were 4, 6, and 9 ml/kg, respectively.The rats in NC and MG were treated with an equal volume of purified water for 10 days.On the third day of pregnancy, except for the NC group, the other groups were injected with mifepristone subcutaneously at the back of the neck at a dose of 5 mg/kg to cause an embryo implantation barrier model.On the 10th day of pregnancy, blood was collected from the abdominal aorta in each group.Serum follicle stimulating hormone (FSH), interferon-γ (IFN-γ) and interleukin (IL-4) were measured by enzyme-linked immunosorbent assay.The number of embryo implantation was observed in the uterus, and the pathological changes of the uterus were observed by HE staining., Results: Compared with the NC group, the number of embryo implantation and the serum levels of FSH and IL-4 in the MG group were decreased significantly ( P < 0.05, 0.01), and pathological changes such as uterine glandular epithelial hyperplasia and inflammatory cell infiltration in the glandular cavity were observed.Compared with MG group, the number of embryo implantation and serum FSH and IL-4 levels of rats in YK-M and YK-H groups were increased significantly ( P <0.05, 0.01).The pathological changes such as uterine glandular epithelial hyperplasia and inflammatory cell infiltration in the gland were also improved.There was no significant difference in serum IFN-γ levels between the groups., Conclusions: Yun Kang oral liquid may improve the endometrial pathological changes and increase the number of embryo implantation by increasing the levels of serum sex hormone FSH and immune cytokine IL-4 in embryo implantation impediment rats.

The synchronization problems of networks of coupled harmonic oscillators are addressed by the edge event-triggered approach in this paper. The network dynamics with respect to edge states are presented and a new edge event-triggered control protocol is designed. Combined with the periodic event-detecting and edge event-triggered approach, sufficient conditions that guarantee the synchronization of coupled harmonic oscillators are presented. Two event-detecting rules are given to achieve the synchronization of coupled harmonic oscillators with low resource consumption. Finally, simulations are conducted to illustrate the effectiveness of the edge event-triggered control algorithm.

In this study, we investigate the keys to obtain a sensitive ammonia sensor with high air stability by using a low-cost polythiophene diode with a vertical channel and a porous top electrode. Poly(3-hexylthiophene) (P3HT) and air-stable poly(5,5'-bis(3-dodecyl-2-thienyl)-2,2'-bithiophene) (PQT-12) are both evaluated as the active sensing layer. Two-dimensional current simulation reveals that the proposed device exhibits numerous connected vertical nanometer junctions (VNJ). Due to the de-doping reaction between ammonia molecules and the bulk current flowing through the vertical channel, both PQT-12 and P3HT VNJ-diodes exhibit detection limits of 50-ppb ammonia. The P3HT VNJ-diode, however, becomes unstable after being stored in air for two days. On the contrary, the PQT-12 VNJ-diode keeps an almost unchanged response to 50-ppb ammonia after being stored in air for 25 days. The improved storage lifetime of an organic-semiconductor-based gas sensor in air is successfully demonstrated.

Next-generation sequencing was used to investigate 9 rare Chinese pedigrees with rare autosomal recessive neurologic Mendelian disorders. Five probands with ataxia-telangectasia and 1 proband with chorea-acanthocytosis were analyzed by targeted gene sequencing. Whole-exome sequencing was used to investigate 3 affected individuals with Joubert syndrome, nemaline myopathy, or spastic ataxia Charlevoix-Saguenay type. A list of known and novel candidate variants was identified for each causative gene. All variants were genetically verified by Sanger sequencing or quantitative polymerase chain reaction with the strategy of disease segregation in related pedigrees and healthy controls. The advantages of using next-generation sequencing to diagnose rare autosomal recessive neurologic Mendelian disorders characterized by genetic and phenotypic heterogeneity are demonstrated. A genetic diagnostic strategy combining the use of targeted gene sequencing and whole-exome sequencing with the aid of next-generation sequencing platforms has shown great promise for improving the diagnosis of neurologic Mendelian disorders., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Genetic heterogeneity is common in many Mendelian disorders such as hereditary spastic paraplegia (HSP), which makes the genetic diagnosis more complicated. The goal of this study was to investigate a Chinese family with recessive hereditary spastic paraplegia, of which causative mutations could not be identified using the conventional PCR-based direct sequencing. Next-generation sequencing of all the transcripts of whole genome exome, after on-array hybrid capture, was performed on two affected male subjects (the proband and his brother). A missense mutation (c.1661G>A, p.R554H) was identified in ABCD1. Subsequently, PCR-based direct sequencing of other family members revealed that the mutation was co-segregating with the disease, indicating that ABCD1 mutation was the pathogenic event for this family. Very long-chain fatty acids (VLCFA) assay in the two affected cases confirmed X-ALD. Our study suggests exome sequencing can be used not only to find a novel causative gene, but also to quickly identify mutations of known genes when the clinical elements are etiologically misleading., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)

Objective: To assess the value of fluorescence in situ hybridization (FISH) combined with chromosomal analysis for the detection of Robertsonian translocation type trisomy 21 in amniotic fluid cells., Methods: Amniotic fluid samples from pregnant women requesting prenatal diagnosis were cultivated. Metaphase cells were prepared for G-banding karyotype analysis. For the 5 Robertsonian translocation type trisomy 21, interphase nuclei from amniotic fluid and parental peripheral blood cells were prepared for FISH analysis., Results: In 2 cases, analysis of parental peripheral blood cells showed normal karyotypes. FISH analysis of amniotic fluid cells indicated that one sample had two copies of chromosome 21, which has a 46, XY, rob(21;21)(q10;q10) karyotype, whilst another had trisomy 21 by FISH, which has a 46, XY, rob(14;21)(q10;q10) karyotype. For the remaining three samples, analysis of parental peripheral blood cells indicated that their karyotypes were 45, XX, rob(14;21)(q10;q10), 45, XX, rob(15;21)(q10;q10) and 45, XX, rob(21;22)(q10;q10), whilst the karyotypes of amniotic fluid cells were 46, XX, rob(14;21)(q10;q10), 46, XY, rob(15;21)(q10;q10) and 46, XX, rob(21;22)(q10;q10), respectively., Conclusion: Combined FISH and chromosomal analysis is an efficient method for detecting non-homologous Robertsonian translocation type trisomy 21. However, FISH has limited ability to detect homologous Robertsonian translocation type trisomy 21.

We successfully demonstrate the first solid-state sensor to have reliable responses to breath ammonia of rat. For thioacetamide (TAA)-induced hepatopathy rats, we observe that the proposed sensor can detect liver that undergoes acute-moderate hepatopathy with a p-value less than 0.05. The proposed sensor is an organic diode with vertical nanojunctions produced by using low-cost colloidal lithography. Its simple structure and low production cost facilitates the development of point-of-care technology. We also anticipate that the study is a starting point for investigating sophisticated breath-ammonia-related disease models.

Interleukin-1 receptor antagonist (IL-1Ra) has been supposed to play important roles in pregnancy. The purpose of this study was to evaluate the association between the polymorphisms of IL-1Ra gene (IL1RN) variable number tandem repeat (VNTR) in intron 2 with idiopathic recurrent spontaneous abortion (RSA). Ninety-two RSA patients and hundred normal women with at least one live birth and no history of miscarriage were included in the study. Frequencies of the IL1RN alleles and genotypes were determined. Data revealed that the prevalence of IL1RN allele and genotype was not significant between the RSA and control group (all P > 0.05). Our finding indicated that the polymorphism VNTR of IL1RN gene in intron 2 may not be a risk factor for RSA in the Chinese Han population., (© 2010 Blackwell Publishing Ltd.)

Human leukocyte antigen-G (HLA-G) molecule exerts multiple immunoregulatory functions that have been suggested to contribute to the immune evasion of tumour cells. Studies on HLA-G expression in malignant haematopoietic diseases are controversial, and the functions of HLA-G on this context are limited. In the current study, HLA-G expression was analysed in different types of patients: de novo acute myeloid leukaemia (AML, n = 54), B cell acute lymphoblastic leukaemia (B-ALL, n= 13), chronic myeloid leukaemia (CML, n= 9) and myelodysplastic syndrome (MDS, n= 11). HLA-G expression was observed in 18.5% cases of AML, 22.2% in CML and 18.2% in MDS, but not in B-ALL patients. In AML, HLA-G-positive patients had a significant higher bone marrow leukaemic blast cell percentage when compared with that of HLA-G-negative patients (P < 0.01). Total T-cell percentage was dramatically decreased in HLA-G-positive patients (P < 0.05). Cytogenetic karyotyping results showed that all HLA-G-positive AML patients (n= 5) were cytogenetically abnormal, which was markedly different from that of HLA-G-negative patients (P < 0.01). Ex vivo cytotoxicity analysis revealed that HLA-G expression in AML leukaemic cells could directly inhibit NK cell cytolysis (P < 0.01). These findings indicated that HLA-G expression in AML is of unfavourable clinical implications, and that HLA-G could be a potential target for therapy.

Amniotic fluid cells (AFCs) are routinely obtained and expanded in vitro for prenatal diagnosis; nevertheless current knowledge about their properties is limited. The detailed mechanisms underlying normal pregnancies are yet to be discovered. The goal of this study was to identify the immunological aspects of AFCs including cytokine production and human leukocyte antigen (HLA) expression, and to discuss its implication for pregnancy. Eighty-six samples of AFCs were determined for HLA expression before and after culture. Cytokine production was measured with flow cytometry in AFC culture supernatants. Treatment of interferon (IFN)-gamma on induction of HLA-DR expression in cultured AFCs was also investigated. Data indicated that both fresh and cultured AFCs express HLA-I, HLA-G, but not HLA-DR, and the cultured AFCs predominately produce the cytokine interleukin (IL)-6. Importantly, we observed that IFN-gamma could induce HLA-DR expression on cultured AFCs in a dose-dependent manner. Taken together, our results indicated that AFCs are functionally active cells and are significant in pregnancy.

Problem: To investigate possible roles of the natural killer (NK) cell receptor killer immunoglobulin-like receptor (KIR)2DL4 expressed on uterine NK (uNK) cells during pregnancy, we investigated KIR2DL4 expression on uNK cells isolated from patients with early recurrent spontaneous abortion (RSA) and normal early pregnancy women, and functions of KIR2DL4 was analyzed in vitro. METHODS OF THE STUDY: Semi-quantitative RT-PCR analysis was introduced to detect KIR2DL4 messenger RNA (mRNA) expression on uNK cells. Cytotoxicity and cytokine production as the result of interaction of KIR2DL4 and its ligand human leukocyte antigen (HLA)-G were analyzed in vitro with lactic dehydrogenase releasing method and enzyme-linked immunosorbent assay, respectively., Results: No significant difference in KIR2DL4 mRNA expression was observed, while the KIR2DL4 protein level in isolated uNK cells is much higher in normal controls than that in RSA patients. Data showed that HLA-G transfection could not reverse the lysis of uNK against HLA-G transfected K562 cells but induced cytokine production. Furthermore, we demonstrated that, via KIR2DL4, membrane-bound HLA-G could induce high cytotoxicity and cytokine production in a high cytotoxic, IL-2 dependent human NK cell line NK-92 cells., Conclusion: Our data suggest that KIR2DL4 might play a crucial implication for human pregnancy.

Human leukocyte antigen (HLA)-G has been postulated as an important immunotolerant molecule in maintaining fetal-maternal relationship. Recent reports indicated that the 14-bp deletion/insertion polymorphism in exon 8 of HLA-G gene influences HLA-G mRNA stability and isoform splicing patterns, thus modulating the levels of HLA-G expression. This might play an immunomodulatory role of HLA-G during implantation and pregnancy. In the present study, 109 unrelated fertile control women and 79 women who had experienced recurrent spontaneous abortion (RSA) were genotyped for the 14-bp insertion/deletion polymorphism. No significant difference was observed in the distribution of 14-bp insertion/deletion genotype between controls and the RSA group. However, a greater number of 14-bp insertion alleles exist in the RSA group than in the controls.

Pre-eclampsia is a multisystem disorder of pregnancy and remains the leading cause of both maternal and fetal morbidity and mortality in many countries. Despite extensive studies, the underlying mechanisms still remain unknown. Besides its restricted expression in the tissues of placenta and its function in regulating immune suppression and in ensuring successful invasion of placental tissues into maternal deciduas, it has been postulated that HLA-G may play a role in modulation of immune tolerance at the fetal-maternal interface. Aberrant HLA-G expression may result in pregnancy disorders that are associated with poor invasion of extravillous cytotrophoblast into maternal spiral arteries, such as pre-eclampsia. Studies have shown that pre-eclampsia is largely under genetic control, but genetic mechanisms underlying the disorder have yet to be determined. In the current study, we focus on the potential role of HLA-G polymorphism in the pathogenesis of pre-eclampsia. Samples were obtained from Chinese Han primiparous women with pre-eclampsia and irrelative normal women, and case-matched placentas were genotyped for the HLA-G polymorphism in the exons 2, 3, and 4, and the 14-base-pair (bp) insertion/deletion polymorphism in the 3'-untranslated region of exon 8 was analyzed separately. The frequency of HLA-G polymorphism in these samples was not significantly different from those of normal controls, indicating that maternal HLA-G polymorphism is not associated with the risk for pre-eclampsia in this Chinese Han population. However, the maternal 14-bp insertion/deletion polymorphism is ethnically different.

In order to study the influence of trichosanthin (TCS) on apoptosis and growth inhibition of human NB4 cells in vitro, the expression of annexin V and the change of DeltaPsim of NB4 cells induced by TCS was analyzed by FACS, and MTT assay was adopted to measure the growth inhibition ratio of NB4 cells treated with TCS. Apoptosis was assayed by agarose gel electrophoresis. The results showed the higher concentration of TCS and the longer the acting time, the stronger growth inhibition of NB4 cells. The expression of annexin V was positive, and the positive ratio was greatly enhanced with prolongation of acting time. DeltaPsim reduced gradually while the apoptosis cells increasing. DNA agarose gel electrophoresis showed a gradient, which confirmed that TCS could induce NB4 cells apoptosis. In conclusion, taken together, data show that TCS can inhibit NB4 growth in vitro, and induce apoptosis. Experiment provides an important evidence for application of TCS in clinical treatment of acute promyelocytic leukemia.

Epidermal growth factors receptor (EGFR) was localized immunocytochemically in the testes of mature and immature rats and immature monkeys. One polyclonal antibody, recognizing the intracellular domain (RK2) of the receptor, was used to carry out the EGFR immunodetection. The RK2 antibody revealed the presence of the EGFR predominantly in Sertoli cells of mature and immature rats and of immature monkeys, although limited interstitial localization of the EGFR was also discerned in the mature rat. In cultured Sertoli cells of immature rats, grown in the absence of epidermal growth factor (EGF), the EGFR was randomly distributed at the cell surface, whereas after the addition of EGF the receptor became aggregated into distinct focal regions. In addition, EGFR of cultured Sertoli cells exhibited autophosphorylation activity upon stimulation with EGF, but failed to transcytose iodinated EGF across a permeability barrier formed by the cultured cells. Instead, all of the added iodinated EGF was internalized and degraded.

We have examined the early events of cellular attachment and spreading (10-30 min) by allowing chick embryonic fibroblasts transformed by Rous sarcoma virus to interact with fibronectin immobilized on matrix beads. The binding activity of cells to fibronectin beads was sensitive to both the mAb JG22E and the GRGDS peptide, which inhibit the interaction between integrin and fibronectin. The precise distribution of cytoskeleton components and integrin was determined by immunocytochemistry of frozen thin sections. In suspended cells, the distribution of talin was diffuse in the cytoplasm and integrin was localized at the cell surface. Within 10 min after binding of cells and fibronectin beads at 22 degrees C or 37 degrees C, integrin and talin aggregated at the membrane adjacent to the site of bead attachment. In addition, an internal pool of integrin-positive vesicles accumulated. The mAb ES238 directed against the extracellular domain of the avian beta 1 integrin subunit, when coupled to beads, also induced the aggregation of talin at the membrane, whereas ES186 directed against the intracellular domain of the beta 1 integrin subunit did not. Cells attached and spread on Con A beads, but neither integrin nor talin aggregated at the membrane. After 30 min, when many of the cells were at a more advanced stage of spreading around beads or phagocytosing beads, alpha-actinin and actin, but not vinculin, form distinctive aggregates at sites along membranes associated with either fibronectin or Con A beads. Normal cells also rapidly formed aggregates of integrin and talin after binding to immobilized fibronectin in a manner that was similar to the transformed cells, suggesting that the aggregation process is not dependent upon activity of the pp60v-src tyrosine kinase. Thus, the binding of cells to immobilized fibronectin caused integrin-talin coaggregation at the sites of membrane-ECM contact, which can initiate the cytoskeletal events necessary for cell adhesion and spreading.

Anordrin has been used as an effective postcoital contraceptive in China. The mechanism of anordrin and its analogue SIPPR-113 on antifertility has been studied. Anordrin and SIPPR-113 possessed estrogenicities and induced decrease in serum progesterone levels in rats. Their antiprogesterone activities might be mainly caused by their estrogenicities, which were the main but not the only contributors for the antifertility. The direct effects of anordrin and SIPPR-113 on human trophoblast cells were studied. A concentration of 50 micrograms/ml or 100 micrograms/ml of anordrin or SIPPR-113 could injure the human trophoblast cells in vitro. The uterine Pontamine blue reaction of mated rats was inhibited in those treated with anordrin or SIPPR-113 at the dose of 4 mg/kg. Anordrin, SIPPR-113 or AF-45 was given orally, intramuscularly and intravenously. The effects of drugs administered via the three routes were nearly the same. This study further demonstrated that anordrin was hydrolyzed to break its bond of dipropionate and was transformed into its parent steroid AF-45 to exert its antifertility effects in vivo. This study warrants that anordrin should been evaluated further.