Gene testing for osteonecrosis of the femoral head in systemic lupus erythematosus using targeted next-generation sequencing: A pilot study.

BACKGROUND: Previous publications indicated that genetic predisposition might play important roles in the onset of osteonecrosis of the femoral head (ONFH) in systemic lupus erythematosus (SLE). Some gene loci such as complement C3d receptor 2 (CR2), nitric oxide synthase 3 (NOS3), collagen type II alpha 1 chain (COL2A1), protein tyrosine phosphatase non-receptor type 22 (PTPN22), and transient receptor potential cation channel subfamily V member 4 (TRPV4) were reported to be involved in this process. AIM: To investigate whether the risk of ONFH in SLE is associated with single nucleotide variations (SNVs) in these five genes. METHODS: SNVs in the CR2, NOS3, COL2A1, PTPN22, and TRPV4 genes were examined by using FastTarget and Illumina Miseq sequencing technologies in 49 cases of SLE with ONFH. Burrows-wheeler aligner was used to align the sequencing reads to hg19, and GATK and Varscan programs were used to perform SNV calling. PolyPhen-2, SIFT, and MutationTaster were used to assess the functional effects of non-synonymous SNVs. RESULTS: Six of the 49 patients were confirmed to have low frequency SNVs, including one patient with SNVs in NOS3 (exon 6: c.814G>A: p.E272K and exon 7: c.814G>A: p.E272K.), four in COL2A1 (rs41263847: exon 29: c.1913C>T: p.T638I, exon 28: c.1706C>T: p.T569I, and rs371445823: exon 8: c.580G>A: p.A194T, exon 7: c.373G>A: p.A125T), and one in CR2 (rs45573035: exon 2: c.200C>G: p.T67S). CONCLUSION: The onset of ONFH in SLE might be associated with the identified SNVs in NOS3, COL2A1, and CR2.

miR­222 regulates brain injury and inflammation following intracerebral hemorrhage by targeting ITGB8.

Intracerebral hemorrhage (ICH) is a disease associated with high mortality and morbidity. MicroRNAs (miRNAs) have been reported to be associated with the pathogenesis of numerous cerebrovascular diseases, including ICH. miR­222 has been revealed to play important roles in various physiological and pathological processes in cardiovascular diseases. However, its role in ICH remains largely unknown. The aim of the present study was to evaluate the potential effect of miR­222 on brain injury in ICH. The results revealed that the expression of miR­222 was significantly increased in ICH, and downregulation of miR­222 significantly reduced erythrocyte lysate­induced cell apoptosis by decreasing the levels of cleaved caspase­3, cleaved caspase­9 and Bax and increasing the level of Bcl­2. In addition, downregulation of miR­222 suppressed the inflammatory responses in erythrocyte lysate­induced microglia, and inhibited inflammation, brain water content and improved neurological functions in ICH mice. Mechanistically, integrin subunit ß8 (ITGB8) was identified as a direct target of negative regulation by miR­222 in microglia cells, and up­regulation of ITGB8 led to the attenuation of inflammation and apoptosis. Collectively, the present findings indicated that miR­222 was a crucial regulator of inflammation via targeting of ITGB8, and represented a promising therapeutic strategy for ICH.

Application of intrapulmonary wire combined with intrapleural fibrin glue in preoperative localization of small pulmonary nodules.

OBJECTIVE: This study aims to investigate the accuracy of the preoperative localization of small nodules by computerized tomography (CT)-guided placing wire and intrapleural fibrin glue near the nodules at 3 days before the operation. METHODS: From October 2015 to December 2017, a total of 79 patients, who received preoperative localization of small pulmonary nodules and surgical treatment in the Department of Thoracic Surgery of Hohhot First Hospital, were enrolled into this study. These patients were randomly divided into 2 groups: methylene blue localization group (n = 47), and modified localization group (n = 32), where the patients received preoperative localization of the small nodules by CT-guided placing wire and intrapleural fibrin glue near the nodule at 3 days before the operation. Localization accuracy, operation time and difficulty in postoperative seeking for pathological specimens were compared between these 2 groups. RESULTS: In the methylene blue localization group, 3 patients had localization failure due to the intrathoracic diffusion of methylene blue, and the success rate was 93.61%. In the modified localization group, all 32 patients succeeded in the localization, and the success rate was 100%. Operation time and difficulty of finding the specimen was significantly lower in the modified localization group than in the methylene blue localization group (P < .05). CONCLUSION: The application of preoperative localization of small nodules by placing wire and intrapleural fibrin glue improves the success rate of resection, reduces operation time and the risk of the operation, and lowers the difficulty of finding pathological specimens after the operation. Hence this operative procedure is worthy of popularization.

Disease-association of different killer cell immunoglobulin-like receptors (KIR) and HLA-C gene combinations in reactive arthritis.

BACKGROUND: Reactive arthritis (ReA) is sterile arthritis triggered by bacterial gastrointestinal or urogenital infections. Although the pathogenesis of ReA remains unclear, genetic factors seem to play an important role. Different killer cell immunoglobulin-like receptors (KIRs) and their corresponding specific histocompatibility leukocyte antigen-C (HLA-C) ligand genotypes have been implicated in susceptibility and resistance to infections and autoimmune diseases but have, thus far, not been investigated in ReA. METHODS: This study was conducted in 138 ReA patients (65 females, 73 males); aged 18-69 years (mean, 37 years) and 151 randomly selected healthy control individuals matched for ethnicity, age and sex. These subjects were genotyped for KIR genes and HLA-C alleles by polymerase chain reaction with sequence-specific primers. RESULTS: The frequencies of inhibitory KIR2DL2 and KIR2DL5 were significantly lower in the ReA patients than in the controls (p = .005 and p = .033, respectively). The presence of more than seven inhibitory KIR genes was protective (p = .016). Moreover, we found that activating KIR2DS1 alone or in combination with the HLA-C1C1 genotype (which indicates the absence of the HLA ligands for their homologous inhibitory receptor KIR2DL1) is associated with susceptibility to ReA (p = .039 and p = .011, respectively), whereas KIR2DL2 in combination with the HLA-C1 ligand is associated with protection against ReA (p = .039). CONCLUSION: These observations indicate that high levels of activating and low levels of inhibitory KIR signals may affect the functions of NK cells and T cells. This imbalance enables the innate and adaptive immune responses of the host to be easily triggered by pathogens, resulting in the overproduction of local and systemic cytokines that contribute to the pathogenesis of ReA.

Antitumor Compounds from the Stout Camphor Mushroom Taiwanofungus camphoratus (Higher Basidiomycetes) Spent Culture Broth.

A known compound, 5-(hydroxymethyl) furan-2-carbaldehyde, and a novel compound, 3-isobutyl-1-methoxy-4-(4'-(3-methylbut-2-enyloxy)phenyl)-1H-pyrrole-2,5-dione were isolated from spent broth from submerged cultures of Taiwanofungus camphoratus. Their structures were elucidated by nuclear magnetic resonance (1H, 13C, and 2D) and mass spectra. These compounds inhibited the proliferation of K562 and HepG2 tumor cells in vitro.

[Study of three ciguatera fish poisoning cases in Xiamen city, in 2005].

OBJECTIVE: To find out the reason of three ciguatera fish poisoning cases in Xiamen in 2005 and identify the fish species. METHODS: The grouper implicated in food poisoning and seven other coral reef fishes collected from market were tested by mice bioassay and ciguatoxin-test kit. The mtDNA was extracted from toxic grouper meat, and Cty b gene segment was amplified and the PCR products were sequenced. The sequences were compared with those in the GenBank. RESULTS: The result turned out to be positive by the ciguatoxin-test kit, while the toxicity of the toxic grouper implicated in food poisoning was 0.11 mouse unit (MU)/g by mice bioassay. A 475 bp segments of Cty b gene was amplified by PCR and the sequence was 99% homologous with Epinephelus fuscoguttatus (GenBank: AY950695).No ciguatoxin in six grouper species collected from market was detected. CONCLUSION: All three food poisoning cases were caused by consumption of ciguatoxin-carrying groupers.

Generation and characterization of antibody against paf1 complex in Drosophila melanogaster.

Paf1 complex was identified in yeast and characterized to function in transcription and its related events. We identified the Drosophila homological components of paf1, CDC73 and RTF1 of paf1 complex. The genes encoding Drosophila paf1, CDC73 and RTF1 were cloned and expressed. With the purified recombinant proteins of truncated components of paf1 complex, antibodies against the Drosophila paf1, CDC73 and RTF1 were generated. These antibodies have been shown to be able to detect the endogenous paf1 subunits as well as their human counterparts in the HeLa extract. On Drosophila polytene chromosomes, these antibodies have been demonstrated to locate the paf1 complex at actively transcribing sites, which co-localized with phosphorylated RNA polymerase II, indicating that paf1 complex in Drosophila is involved in transcription or the events coupling with transcription.

[Effect of anti-Helicobacter pylori ureB monoclonal antibody on platelet aggregation and activation, and its mechanism study].

OBJECTIVES: To study the effect of monoclonal antibody (McAb) against helicobacter pylori (Hp) ureB, 1F11 on platelet aggregation and activation, and its mechanism. METHODS: The relativity between human platelet glycoproteins (GPs) and Hp ureB was identified by Western blot and FCM. Platelet aggregation was measured by turbidimetry, and P-selectin and TXB2 assay by ELISA. RESULTS: 1F11 could bind to platelet GPIIIa, and ADP-induced platelet aggregation was inhibited by 1F11 in a dose-dependent manner. However, 1F11 had no effect on plasma P-selectin and TXB2 induced by ADP. The FCM results show that the positive rates of platelet binding to FITC-SZ21 was decreased from 99.5% to 77.4% after addition of 1F11. CONCLUSION: McAb against Hp ureB 1F11 inhibits platelet aggregation through binding to platelet GPIIIa but does not block platelet activation. There might be crossed-epitopes on Hp ureB and platelet GPIIIa, and Hp infection might be involved in ITP immunopathology.