This study is to explore the interventional effects of fluvastatin on anti-beta2GPI/beta2GPI-induced activation in THP-1 mononuclear cells. In vitro, human mononuclear cells THP-1 were treated with fluvastatin, LPS and anti-beta2GPI/beta2GPI, then the TF expression on THP-1 cells was detected by real-time quantitative PCR (RT-qPCR) or TF activity was detected by kit. TNF-alpha mRNA and its protein expression were investigated by RT-PCR and ELISA kit. The expression of phospho-NF-kappaB p65 and inhibitory protein of NF-kappaB (IkappaB-alpha) were measured by Western blotting. The results suggested that the expression of TF and TNF-alpha on THP-1 cells was significantly up-regulated with treatment of anti-beta2GPI/beta2GPI complex (100 mg x L(-1)), compared with that of untreated cells (P < 0.05). Fluvastatin (50 mg x L(-1)) could decrease TF (mRNA and activity) expression and the level of TNF-alpha (mRNA and protein) in THP-1 cells with anti-beta2GPI/beta2GPI complex. The expression of TF and TNF-alpha was shown in a concentration-dependent manner. Moreover, anti-beta2GPI/beta2GPI complex could downregulate IkappaB-alpha levels and increase the levels of phospho-NF-kappaB p65. And these effects of anti-beta2GPI/beta2GPI complex could be blocked by fluvastatin. In conclusion, fluvastatin may interfere the expression and regulation of NF-kappaB signal transduction pathway, thereby inhibit the effects of anti-beta2GPI/beta2GPI on activation of THP-1 cells, by decreasing the expression of TF and TNF-alpha.

Objective To investigate the regulation effect of hepatocyte nuclear factor (HNF) 1α on the gene expression profile and the signal pathways in HuH7 cells.Methods The expression of HNF1α was increased or decreased in HuH7 cells by Lenti-virus carrying HNF1α or shHNF1α.The expression profile of the cells after treated was examined by microarray technology.The difference expressed gene regulated by HNF1α were screened and the pathway was analyzed with DAVID software and related analysis system.The regulation effect of HNF1α on transforming growth factor (TGF)β signal pathway was detected by reporter gene test and the regulation role of HNF1α on related genes of TGFβ signal pathway was determined by real-time polymerase chain reaction (PCR) and Western blotting assay.Results The expression of HNF1α in HuH7 cells was significantly up-regulated by Lenti-virus carrying HNF1α gene (Lenti-HNF1α) and which was down regulated by Lenti-virus with shHNF1α gene (LentishHNF1 α).Expression profile analysis revealed that 339 genes were positively up regulated two times by HNF1α and 325 genes were negatively down regulated two times.Signal pathway analysis revealed that HNF1α regulated drug metabolism,biosynthesis of unsaturated fatty acids and glycolysis/gluconeogenesis metabolism signal pathways.Moreover,it also involved in the regulation of TGFβ、nuclear factor (NF)-κB and p53 tumor-related signal pathways.Furthermore,Luciferase reportor gene experiment indicated that up-regulated HNF1α could inhibit the activation of TGFβ signal pathway.And the results of real-time PCR and Western blotting verified that up-regulated HNF1α could inhibit TGFβ signal pathway related gene c-myc and TGFβ1 and then inhibited the activation of TGFβ signal pathway.Conclusion HNF1α broadly affects the gene expression profile and the tumor genesis and development related signal pathways in HuH7 cells,furthermore,HNF1α can inhibit the activation of TGFβ signal pathway.

Objective To investigate the feasibility of our new found 3-dimensional contrast-enhanced ultrasound 3D-CEUS registration system as an early assessment of the therapeutic response to radio frequency ablation for liver cancer Methods Twenty-seven patients with 28 lesions accepted 3D-CEUS before and after radio frequency ablation RFA the therapeutic respond to which would be assessed with 3D-CEUS registration system recording the rate of successful registration The CT was considered as the reference standard Results Ten cases 35 7% were successful matched with auto-registration and 24 cases 85 7% were succesful matched with interactive-registration relatively All cases were considered as complete ablated which were confirmed by CECT with 100% accuracy There were two cases achieving ablation margins ≥5 mm without local tumor progression LTP and nineteen cases achieving 0 -4 mm ablation margin with 3 LTP 3-month 6-month and 1-year later Conclusions The 3D-CEUS interactive-registration system can easily assess the therapeutic response of RFA in liver cancer immediately with high accuracy.

Objective:To investigate the expression and distribution of human dermal papillary fibroblasts (Fp) , reticular fibroblasts (Fr) , and myofibroblasts (MFB) in keloid tissues.Methods:Keloid tissues were collected from 15 outpatients (including 8 males and 7 females) aged 20-50 years, who were diagnosed in the Department of Dermatology, Renmin Hospital of Wuhan University from May to December 2019. Normal skin tissues were taken from 15 age-matched women who underwent mammoplasty, and served as controls. The distribution of fibroblast activation protein (FAP) , CD90 and alpha-smooth muscle actin (α-SMA) was observed in the keloid tissues and normal skin tissues by dual immunofluorescence staining. Furthermore, fibroblasts were isolated from 3 normal skin and 3 keloid tissue samples, and subjected to primary culture. Subsequently, the fibroblasts were treated with 10 ng/ml transforming growth factor-β1 (TGF-β1) for 48 hours in vitro, during which, changes in fibroblast phenotypes were observed in the 2 groups. Fluorescence-based quantitative RT-PCR and Western blot analysis were performed to determine the mRNA and protein expression of FAP, CD90 and α-SMA. Measurement data were compared between 2 groups by using t test. Results:Immunofluorescence staining of the normal skin tissues revealed that FAP +/CD90 - fibroblasts were predominantly distributed in the superficial dermis, FAP -/CD90 + fibroblasts in the deep dermis, and CD90 + cells hardly expressed α-SMA; however, a large number of FAP + fibroblasts and CD90 + fibroblasts were observed in the deep keloid tissues, and many CD90 + fibroblasts also expressed α-SMA. Dual immunofluorescence staining showed that normal tissue-derived fibroblasts hardly expressed α-SMA, and keloid-derived fibroblasts expressed α-SMA. The fluorescence intensity of α-SMA + cells significantly increased in the normal tissue-and keloid-derived fibroblasts after 24-hour treatment with TGF-β1 (21.058 ± 0.709, 27.112 ± 0.097, respectively) compared with that in the corresponding untreated fibroblasts (11.312 ± 0.636, 21.306 ± 0.464, t=22.430, 13.370, respectively, both P < 0.05) . RT-PCR and Western blot analysis showed that the mRNA and protein expression of FAP, CD90 and α-SMA significantly increased in the keloid-derived fibroblasts after 48-hour treatment with TGF-β1 (mRNA: 92.610 ± 3.667, 1.366 ± 0.105, 3.240 ± 0.141; protein: 0.652 ± 0.073, 1.046 ± 0.119, 0.946 ± 0.117, respectively) compared with the untreated keloid-derived fibroblasts (all P < 0.05) . Conclusion:CD90 + Fr aberrantly proliferated in the deep dermis of keloid tissues, suggesting that directional intervention in aberrantly proliferating FAP -/CD90 + Fr in the deep dermis may promote the efficacy for keloids.

To prepare recombinant fox growth hormone (fGH), we amplified its cDNA from silver fox pituitary tissue by RT-PCR and cloned into yeast shuttle vector pPIC9K down stream of a-factor signal peptide sequence by SnaB I and Not I restriction sites. The recombinant secretion vector pPIC9K/fGH, linearized by Sal I, was transformed into histidine-deficient Pichia pastoris strain GS115 by electroporation. We selected His+ -transformed methylotropic (His+, Mut+) yeast using histidine-absent medium containing dextrose (MD) or methanol (MM) as the only carbon source, and then screened the recombinant GS115 with multi-copy fGH genes by G418. The secretive expression of fGH was performed under the induction of methanol in shaking flask culture. The results showed that the fGH cDNA sequence amplified in this paper was basically in consistence with the published in GenBank. We achieved the secretive expression of recombinant fGH identified by SDS-PAGE and Western blotting. The fGH expression level was 119 mg/L, accounted for 34% of total proteins in fermentation medium.
Animals ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Electroporation ; Foxes ; genetics ; Genetic Vectors ; genetics ; Growth Hormone ; biosynthesis ; genetics ; Molecular Sequence Data ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics

Objective:To preliminarily explore the clinical value of three-dimensional ultrasound fusion imaging(3DUS FI) visualization technology in guiding precise needle placement during thermal ablation of hepatocellular carcinoma (HCC).Methods:A total of 56 HCC patients (59 lesions)who underwent 3DUS FI guided thermal ablation were retrospectively analyzed in the First Affiliated Hospital of Sun Yat-sen University from November 2019 to December 2021. All patients were collected with three-dimensional ultrasound volume image before ablation which were fused with real-time two-dimensional ultrasound image for registration, and then the tumor and the safety margin of 5 mm were segmented and marked. Finally, the thermal ablation was performed under three-dimensional visualization. Contrast-enhanced CT/MRI was performed 1 month after thermal ablation to evaluate whether the lesion was completely ablated and measure the ablative margin, and the relationship between ablative margin and the incidence of local tumor progression (LTP) was also analyzed.Results:During the ablation, all lesions could be successfully registered and displayed in three-dimension. Postoperative contrast-enhanced ultrasound showed that all lesions were completely ablated. A total of 37 lesions could be evaluated for ablative efficacy and ablative margin based on contrast-enhanced CT/MRI 1 month after themal ablation, of which 32 (86.5%) lesions achieved complete ablation and obtained at least 5 mm ablative margin. During the follow-up period, LTP was occurred in 4 lesions, 3 of the lesions occurred at the ablative margin< 5 mm. Both 1-year and 2-year cumulative LTP rates were all 7.1%. None of patients had serious complications or deaths associated with thermal ablation.Conclusions:3DUS FI real-time guidance technology is feasible and safe in visually guiding precise needle placement during thermal ablation of HCC.

Objective To compare three-dimensional contrast-enhanced ultrasound ( 3DCEUS) fusion imaging and computed tomography ( CT ) fusion imaging in evaluating ablation margin ( AM ) after radiofrequency ablation ( RFA) for hepatocellular carcinoma ( HCC) . Methods The 3DCEUS images of 60 patients before and after RFA were collected . The AM was evaluated by the self-developed 3DCEUS fusion imaging technique . The consistency of AM evaluation was compared between 3DCEUS and CT fusion imaging . The risk factors of local tumor progression ( LTP) including AM were analyzed . Results The registration success rate of 3DCEUS fusion imaging was 96 .7％ ( 58/60) . Thirty-one cases were in the AM<5 mm group ,and 27 cases were in the AM ≥5 mm group . The consistency of AM evaluation between 3DCEUS and CT fusion imaging was good ( Kappa coefficient = 0 .895 , P < 0 .001) . During a follow-up period ranging 4 .2 to 18 months ,LTP was identified in 5 tumors (8 .6％ ,5/58) .The incidence of LTP with the AM<5 mm was higher than that with the AM ≥5 mm ( P =0 .033) . Conclusions 3DCEUS fusion imaging is feasible for AM evaluation immediately after RFA with high consistency with CT fusion imaging . AM<5 mm evaluated on 3DCEUS fusion immediately after RFA is a risk factor for LTP .

In order to explore the microbial communities and functions of activated sludge in an Anaerobic-anoxic-oxic (A²/O) process under the start-up of Actinic reaction enzyme system (ARES) system and to understand the impact of the ARES system in domestic sewage treatment process, the activated sludge microbial community structure in the A²/O process system before and after ARES system start-up was analyzed by Illumina-HiSeq 2000 high-throughput sequencing platform. By combining with the main parameters related to the effect of sewage treatment, we analyzed the environmental functions of the microbial communities. The microbial community structure of activated sludge was significantly different before and after the ARES system start-up. There were 9 main bacterial phyla in the system (average relative abundance ≥1%), accounting for 96%-98% of the total bacteria sequenced. After the ARES system was started, the relative abundance of Betaproteobacteria and Chlorobi increased by 3.45%-3.85% and 0.45%-2.61%, respectively. In the anaerobic unit, the relative abundance of Bacteroidetes increased by 12.97%, while the Actinobacteria and Firmicutes decreased by 9.60% and 1.45%, respectively. At the genus level of bacteria, the relative abundance of Denitratisoma increased by 0.80%-3.27%, while the Haliangium and Arcobacter decreased by 3.36%-4.52% and 1.48%-3.45%, respectively. The relative abundance of bacteria was significantly different before and after the ARES system start-up. There were 7 abundant fungi phyla (average relative abundance ≥1%) in the system. After the ARES system was started, the relative abundance of Rozellomycota decreased by 42.71%-46.77%. In the anaerobic unit, the relative abundance of Ascomycota decreased by 13.39%, while the relative abundance of Glomeromycota increased by 13.86%. At the genus level of fungi. The relative abundance of Entomophthoraceae sp. and Glomcromycota sp. increased by 31.35%-36.50% and 6.27%-13.84%, respectively, while the Rozellomycota sp. and Xylochrysis lucida decreased by 42.71%-46.77% and 3.67%-5.54%, respectively. Our results showed that the application of ARES system caused the response of the microbial community to environmental changes, especially for the fungi communities, in the meanwhile, improved the effluent quality, especially the removal rate of total nitrogen.
Anaerobiosis ; Ascomycota ; Bioreactors ; Microbiota ; Nitrogen ; Sewage ; Waste Disposal, Fluid

Lung cancer remains the leading cause of cancer deaths worldwide and is the most common cancer in males. Immune-checkpoint inhibitors (ICIs) that target programmed cell death protein-1 (PD-1) or programmed cell death-ligand 1 (PD-L1) have achieved impressive efficacy in the treatment of non-small-cell lung cancer (NSCLC) (Pardoll, 2012; Champiat et al., 2016; Gao et al., 2022). Although ICIs are usually well tolerated, they are often accompanied by immune-related adverse events (irAEs) (Doroshow et al., 2019). Non-specific activation of the immune system produces off-target immune and inflammatory responses that can affect virtually any organ or system (O'Kane et al., 2017; Puzanov et al., 2017). Compared with adverse events caused by chemotherapy, irAEs are often characterized by delayed onset and prolonged duration and can occur in any organ at any stage of treatment, including after cessation of treatment (Puzanov et al., 2017; von Itzstein et al., 2020). They range from rash, pneumonitis, hypothyroidism, enterocolitis, and autoimmune hepatitis to cardiovascular, hematological, renal, neurological, and ophthalmic irAEs (Nishino et al., 2016; Kumar et al., 2017; Song et al., 2020). Hence, we conducted a retrospective study to identify validated factors that could predict the magnitude of the risk of irAEs in patients receiving PD-1/PD-L1 inhibitors; our approach was to analyze the correlation between the clinical characteristics of patients at the start of treatment and relevant indicators such as hematological indices and the risk of developing irAEs. Then, we developed an economical, practical, rapid, and simple model to assess the risk of irAEs in patients receiving ICI treatment, as early as possible.
Male ; Humans ; Carcinoma, Non-Small-Cell Lung/drug therapy* ; Lung Neoplasms/drug therapy* ; Immune Checkpoint Inhibitors/adverse effects* ; Programmed Cell Death 1 Receptor ; Retrospective Studies ; Apoptosis