OBJECTIVE To investigate the therapeutic effect and side effect of ibrutinib long-term treatment in systemic lupus erythematosus (SLE) model mice induced by pristane. METHODS Female 6-week old BALB/c mice were ip given pristane 0.5 mL once for SLE induction. Four weeks later, the SLE model mice were divided into three groups based on the body mass and serum level of anti-double-strand DNA (DS-DNA) antibody and treated with 1%methylcellulose (model control group, ig), ibrutinib (30 mg·kg-1, ig) or prednisone (10 mg·kg-1, ig), respectively, once daily for 28 weeks. Every 4 weeks, body mass of each mouse was measured. Autoantibodies against DS-DNA, single-strand DNA (SS-DAN), and histone were tested by ELISA. The incidence of lupus arthritis and clinical score of inflammation and edema were recorded and graded. Biochemical analysis of urea protein, serum urea nitrogen and creatinine was used to evaluate kidney function. Mice were euthanized post 28 weeks of dosing. Inter-leukin-6 (IL-6), interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) were tested by ELISA. The heart, liver, spleen, lung, and kidney were collected and weighed for organ relative mass calculation. Biochemical analysis of serum glutamic-pyruvic transaminase (GPT), glutamic-oxal(o)acetic transaminase (GOT) and alkaline phosphatase (ALP) was used to evaluate liver function. Hind feet were collected for HE staining and pathological scoring to observe renal injury and inflammation. Immunohistochemical staining was used to study IgG immune complexes deposition in kidneys of lupus. RESULTS Compared with normal control group, autoantibodies (anti-DS-DNA, anti-SS-DNA and anti-histone antibodies), renal function indexes (serum creatinine, urea nitrogen and urine protein), and cytokines (IL-6, IFN-γ and TNF-α) levels in model group were significantly increased (P<0.01). The model group mice had obvious clinical symptoms of arthritis (P<0.01), serious inflammation cell invasion (P<0.01), and the mass of kidneys and spleen increased significantly (P<0.01). Compared with model group, after 28 weeks of treatment, ibrutinib decreased the level of anti-DS-DNA, anti-SS-DNA and anti-histone antibodies (P<0.01), decreased the lupus arthritis score (P<0.01) and the morbidity of arthritis, reduced the level of cyto-kines IL-6, IFN-γand TNF-α(P<0.01), reduced the level of serum creatinine, serum urea nitrogen and urine protein (P<0.01), improved pathological symptoms of hind feet such as inflammation, cartilage destruction, bone resorption and pannus (P<0.01), alleviated renal tissue inflammatory cell invasion and the immune-complex precipitation, and reduced the mass of organs (spleen and kidneys, P<0.01) and the level of liver function (GPT and ALP, P<0.01). CONCLUSION Long-term treatment with ibrutinib has therapeutic effect on the model mice of SLE, and has no obvious side effect.

Objective To investigate the characteristics and diagnostic value of 18F-fluorodeoxyglucose (FDG) PET/CT in patients with secondary malignant peripheral nerve lesions. Methods 18F-FDG PET/CT studies of 8 cases of secondary malignant peripheral nerve lesions confirmed by histopathology or follow-up were analyzed retrospectively. The maximum standardized uptake value ( SUVmax ) of infiltrating peripheral nerves and contralateral normal peripheral nerves was measured and compared with their morphological appearances on CT. Paired student t-test was performed by SPSS 10.0. Results Twelve secondary malignant peripheral nerve lesions with high 18F-FDG metabolism were found in 8 cases. On PET imaging,the lesions distributed along the neurovascular tissues or intervertebral foramina with appearances resembling those of fibre bundles,radices or nodes on PET but no density differences with the surrounding soft tissue or fat planes on CT. The SUVmax was 6.86 ± 3.87. The contralateral normal peripheral nerves showed no abnormal 18F-FDG uptake with a SUVmax of 1.10 ±0.46,which was significantly different from that of the secondary malignant peripheral nerve lesions (t = 9.231,P ＜ 0.001 ). Conclusion 18 F-FDG PET/CT may be useful in locating the secondary malignant peripheral nerve lesions and in assessing its regional infiltration.

OBJECTIVETo investigate the characteristics of RET/PTC and H47PTEN rearrangement and the association between gene rearrangement and clinicopathological properties of thyroid carcinoma.

METHODSRearrangement of RET/PTC-1, RET/PTC-2, RET/PTC-3, ELKS-RET and H4-PTEN (H4/PTEN and PTEN/H4) was analyzed in 139 thyroid tumor tissues by using RT-PCR and sequencing.

RESULTSTwelve RET/PTC-1, 6 RET/PTC-3, 6 H4/PTEN and 7 PTEN/H4 were detected in 126 papillary thyroid carcinomas. In 3 cases, both RET/PTC and H4-PTEN were identified simultaneously. However, repeated experiments did not give the same results of H4-PTEN rearrangement. The overall frequency of rearrangement was 21.4% (27/126). The patients with gene rearrangement were younger (P = 0.02) and had a higher frequency of lymph node involvement (P = 0.02). High frequency of lateral neck lymph node involvement was detected in RET/PTC positive PTC (P < 0.01). PTEN/H4 rearrangement could also be detected in medullary thyroid carcinoma (2/5).

CONCLUSIONSH4-PTEN rearrangement can occur simultaneously with RET/PTC rearrangement in PTC. High predisposition to gene rearrangement is a characteristic of PTC. The patients of PTC with gene rearrangement are younger and have a higher frequency of lymph node involvement.

Adolescent ; Adult ; Aged ; Carcinoma ; Carcinoma, Papillary ; Child ; Female ; Gene Rearrangement ; Humans ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; PTEN Phosphohydrolase ; genetics ; Protein-Tyrosine Kinases ; genetics ; Thyroid Neoplasms ; genetics ; pathology ; Young Adult

OBJECTIVETo screen and identify differentially expressed genes between a fertile patient and another infertile patient who belonged to a large Chinese pedigree affected with androgen insensitivity syndrome (AIS).

METHODSWe constructed the forward and reversed subtracted libraries using genital skin fibroblasts (GSF), which were obtained from the fertile patient MJ and infertile patient ZGJ, as tester respectively. Candidate clones were screened with colony in situ hybridization, dot blot, and Southern blot analysis step by step and conformed with Northern blot analysis. The potential positive clones were sequenced and the homology of the sequences was analyzed.

RESULTSThe forward and reversed subtracted libraries containing differentially expressed pattern of two GSF cell lines were constructed. Two positive clones identified by Northern blot were obtained in the reversed subtracted library. Eleven candidate clones from the two libraries that failed to hybridize with both RNA populations were obtained simultaneously, which might represent differentially expressed low abundance transcripts. Sequencing results and homology analysis demonstrated that the two positive clones were significantly homologous with the genes of autotaxin-t and calcium binding protein calcyclin (S100A6), respectively.

CONCLUSIONSTwo positive clones and eleven clones showing no hybridization signals may represent differentially expressed genes between the two GSFs. This finding may be useful to elucidate the molecular mechanisms leading to phenotypic variation and preserved fertility of the AIS pedigree.

Androgen-Insensitivity Syndrome ; complications ; genetics ; Blotting, Northern ; Fertility ; genetics ; Fibroblasts ; cytology ; Gene Expression Profiling ; Gene Library ; Genitalia, Male ; cytology ; Humans ; In Vitro Techniques ; Infertility, Male ; etiology ; genetics ; Male ; Nucleic Acid Hybridization ; methods ; Pedigree ; Polymerase Chain Reaction ; Skin ; cytology

Gut microbiome sequencing studies have great potential to translate microbial analysis outcomes into human health research. Sequencing strategies of 16S amplicon and whole-metagenome shotgun (WMS) are two main methods in microbiome research with respective advantages. However, how sample heterogeneity, sequencers and library preparation protocols affect the sequencing reproducibility of gut microbiome needs further investigation. This study aims to provide a reference for the selection of sequencing technologies by comparing differences in microbial composition from different sampling sites. The results of three widely adopted sequencers showed that the technical repetition correlation (r= 0. 94) was high in WMS method, while the biological repetition correlation (r = 0. 69) was low. Bray-Curtis distance identified that dissimilarity from biological replicates was larger than that of technical replicates (P<0. 001). In addition, dissimilarity and specific taxonomic profiles were observed between 16S and WMS datasets. Our results imply that homogenization is a necessary step before sample DNA extraction. The sequencers contributed less to taxonomic variation than the library preparation protocols. We developed an empirical Bayes approach that " borrowed information" in calculations and analyzed batch effect parameters using standardized data and prior distributions of (non-) parameters, which may improve population comparability between 16S and WMS and provide a basis for further application to fusion analysis of published 16S and microbial datasets.