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Resistance to 5-FU reduces its clinical efficacy for the treatment of colorectal cancer. Sphingosine-1-phosphate receptor 2 (S1PR2) has emerged as a potential target to reverse 5-FU-resistance by inhibiting the expression of dihydropyrimidine dehydrogenase (DPD). In this study, 38 novel S1PR2 antagonists based on aryl urea structure were designed and synthesized, and the structure-activity relationship was investigated based on the S1PR2 binding assay. Representative compound

Asymmetric α-regioselective annulation of MBH carbonates with 4-arylmethylisoxazol-5-ones has been developed to afford spirocyclic oxindole derivatives containing three contiguous stereogenic centers and vicinal all-carbon quaternary chiral centers. This reaction exhibits a broad substrate scope and excellent functional group tolerance. Excellent yields with high diastereo- and enantioselectivities were obtained in this efficient organocatalytic reaction.

Predicting protein function is a longstanding challenge that has significant scientific implications. The success of amino acid sequence-based learning methods depends on the relationship between sequence, structure, and function. However, recent advances in AlphaFold have led to highly accurate protein structure data becoming more readily available, prompting a fundamental question:given sufficient experimental and predicted structures, should we use structure-based learning methods instead of sequence-based learning methods for predicting protein function, given the intuition that a protein’s structure has a closer relationship to its function than its amino acid sequence?To answer this question, we explore several key factors that affect function prediction accuracy. Firstly, we learn protein representations using state-of-the-art graph neural networks (GNNs) and compare graph construction(GC) methods at the residue and atomic levels. Secondly, we investigate whether protein structures generated by AlphaFold are as effective as experimental structures for function prediction when protein graphs are used as input. Finally, we compare the accuracy of sequence-only, structure-only, and sequence-structure fusion-based learning methods for predicting protein function. Additionally, we make several observations, provide useful tips, and share code and datasets to encourage further research and enhance reproducibility.

Introduction Tibial plateau fractures are often accompanied with ligamental and meniscal injuries. Among which, the combined existence of Schatzker type IV fracture with anterior cruciate ligament (ACL) avulsion has been reported rarely. The purpose of this study was to determine the injury mechanism of Schatzker type IV fracture with ACL avulsion based on Mimics software. Methods Ninety-nine Schatzker type IV tibial plateau fractures were retrospectively analyzed by quantitative three-dimensional measurements. ACL avulsions were diagnosed through the data of computed tomography and magnetic resonance imaging. We simulated the knee posture when an injury occurred and defined different injury patterns. The chi-square test was used for determining the main mechanism which causes Schatzker type IV fractures associated with ACL avulsions. Results There were more ACL avulsions and more displaced ACL avulsions associated with the knee in flexion in the setting of Schatzker type IV fracture (p Conclusion This study found that a flexed knee at the occurrence of a Schatzker type IV tibial plateau fracture is a high-risk factor for causing associated ACL avulsion and producing more displaced avulsions. Flexion-valgus pattern was the main cause of Schatzker type IV fractures associated with ACL avulsions. The findings will help orthopedists understand the injury mechanism and enhance their awareness of such injuries to avoid unfavorable prognosis.

A highly efficient diastereoselective [4 + 1] cycloaddition of ortho-tosylaminophenyl-substituted p-QMs with 3-chlorooxindoles has been developed to afford 3,2′-pyrrolidinyl spirooxindoles.

A novel and efficient catalyst-free and oxidant-free tandem aza-Mannich/cyclization/aromatization reaction of C,N-cyclic azomethine imines with enamides has been developed for the synthesis of 5,6-dihydropyrazolo[5,1-a]isoquinolines.

PURPOSE CARTIFAN-1 aimed to evaluate the efficacy and safety of ciltacabtagene autoleucel (cilta-cel), a B-cell maturation antigen–targeting chimeric antigen receptor T-cell therapy, in Chinese patients with relapsed/refractory multiple myeloma (RRMM). METHODS This pivotal phase II, open-label study (ClinicalTrials.gov identifier: NCT03758417 ), conducted across eight sites in China, enrolled adult patients with RRMM who had received ≥ 3 lines of prior therapy, including a proteasome inhibitor and immunomodulatory drug. Patients received a single infusion of cilta-cel (target dose 0.75 × 106 chimeric antigen receptor–positive viable T cells/kg). The primary end point was overall response rate. Secondary end points included progression-free survival (PFS), overall survival (OS), and incidence and severity of adverse events (AEs). RESULTS As of the clinical cutoff of July 19, 2021, 48 patients received a cilta-cel infusion. At an 18-month median follow-up, the overall response rate was 89.6% (95% CI, 77.3 to 96.5), with a median time to first response of approximately 1 month; 77.1% of patients (95% CI, 62.7 to 88.0) achieved complete response or better. Medians for duration of response, PFS, and OS were not reached. The 18-month PFS and OS rates were 66.8% (95% CI, 49.4 to 79.4) and 78.7% (95% CI, 64.0 to 88.0), respectively. Hematologic AEs were common, including anemia (100%), neutropenia (97.9%), lymphopenia (95.8%), and thrombocytopenia (87.5%). Cytokine release syndrome occurred in 97.9% of patients (35.4% grade 3/4); the median time to onset was 7 days, and the median duration was 5 days. Infections occurred in 85.4% of patients (37.5% grade 3/4). Ten deaths occurred after cilta-cel infusion, eight of which were due to treatment-related AEs. CONCLUSION These data demonstrate a favorable risk-benefit profile for a single infusion of cilta-cel, resulting in early, deep, and durable responses in heavily pretreated patients with RRMM in China.

Background The left ventricular ejection fraction (LVEF) plays a pivotal role in the diagnosis and prediction of ischemic heart disease (IHD). Current techniques to measure LVEF have led to some complications and are relatively expensive despite the high accuracy. Heart rate variability (HRV) is an alternative for the assessment of cardiac function and its related parameters are easily to be derived from electrocardiography. Objective This study aimed to investigate the corresponding relationship between LVEF and HRV in patients with IHD via prediction models developed by machine learning techniques and compare the accuracy of these models. Methods In this retrospective real-world study, patients with IHD admitted to Jinan 4th People’s hospital between January 2019 and December 2020 were randomly selected and divided into a derivation cohort and a validation cohort in a 7:3 ratio. LVEF were measured by color Doppler echocardiography while HRV-related parameters were derived from a 24h Holter electrocardiography recordings. Patient clinical features and HRV parameters were identified and used in the subsequent modelling process. Logistic regression, naïve Bayes, decision tree, gradient boosting, extreme gradient boosting (XGBoosting) and random forests were respectively applied to estimate the correlation between HRV and LVEF. Model performances were evaluated by the coefficient R2 (the closer the result is to 1, the better) and the accuracy (a precise prediction was defined as the accuracy ≥ 90%). Results A total of 179 patients were included in this study with 125 (mean age 67.03 ± 16.12, male 72 [53.7%]) in the derivation cohort and 54 (mean age 62.51 ± 12.97, male 19 [48.7%]) in the validation cohort. Six variables including sex, age, SDNN, SDANN, rMSSD and pNN50 were found to be correlated with LVEF. Gradient boosting and random forests outperformed other machine learning models, showing the best prediction performance (> 90%) in the prediction of both LVEF percentage (%) and LVEF stratifications ( 50%). These six indicators showed different contributions in different algorithms with sex as the least important indicator. Conclusion Gradient boosting and random forests machine learning models based on HRV- and patient characteristics-related parameters provided a new method in predicting LVEF in patients with IHD, which showed a potential in the diagnosis of IHD.

Two-dimensional (2D) transition metal dichalcogenides (TMDs) are ideal elements for many optoelectronic devices owing to their outstanding optoelectrical performance under visible and infrared light. Heterostructures composed of TMDs and other non-TMD materials may exhibit rich properties. In this study, a high-performance heterojunction based on 2D MoTe2 and 2D electron gas (2DEG) at the LaAlO3/SrTiO3 interface was fabricated. The device exhibits good current rectification properties with a high rectification ratio exceeding 103 and a low leakage current (∼1 nA at −6 V bias). Moreover, a high photoresponsivity of ∼800 A W−1 and a large specific detectivity of 4 × 1012 Jones at 405 nm were also obtained at room temperature. Heterostructures based on 2D TMDs and oxide 2DEG are expected to become essential elements in multifunctional microdevices and optoelectronic devices.

The association between ambient airborne fine particulate matter (PM2.5) exposure and respiratory diseases has been investigated in epidemiological studies. To explore the potential mechanism of PM2.5-induced pulmonary fibrosis, 60 mice were divided into 3 groups to expose to different levels of PM2.5 for 8 and 16 weeks: filtered air, unfiltered air, and concentrated PM2.5 air, respectively. BEAS-2B cells were treated with 0, 25, 50, and 100 μg/ml PM2.5 for 24 h. The biomarkers of pulmonary fibrosis, epithelial-mesenchymal transition, N6-methyladenosine (m6A) modification, and metabolism of mRNAs were detected to characterize the effect of PM2.5 exposure. The results illustrated that PM2.5 exposure induced pathological alteration and pulmonary fibrosis in mice. The expression of E-cadherin was decreased whereas vimentin and N-cadherin expression were increased in a dose- and time-dependent manner after PM2.5 exposure. Mechanistically, PM2.5 exposure increased the levels of METTL3-mediated m6A modification of CDH1 mRNA. As a target gene of miR-494-3p, YTHDF2 was upregulated by miR-494-3p down-regulation and then recognized m6A-modified CDH1 mRNA to inhibit the E-cad expression, consequently induced the EMT progression after PM2.5 exposure. Our study indicated that PM2.5 exposure triggered EMT progression to promote the pulmonary fibrosis via miR-494-3p/YTHDF2 recognized and METTL3 mediated m6A modification.

The hybrid integration of oxide two-dimensional electron gases (2DEGs) and 2D layered transition-metal dichalcogenides is expected to exhibit diverse physical phenomena, which cannot appear in either material alone. Here, we demonstrate a gate-free tunable 2D/2D heterojunction composed of MoTe2 and 2DEG at the surface of KTaO3. The combination of metallic MoTe2 and n-type 2DEG together forms a Schottky diode with a large on/off current ratio of 104 at room temperature. Moreover, the MoTe2/2DEG diode exhibits largely tunable electrical transport characteristics without gate voltages. By applying bias voltages, the diode shows tunable transport properties ranging from insulating to excellent rectifying behaviors. The bias-voltage-dependent modulation dominantly originates from the tunable Schottky barrier width controlled by the carrier density of the 2DEG. Our results pave a way for the development of 2D nanoelectronic devices such as multi-bit memories and bias sensors.

5-Fluorouracil (5-FU) and its prodrugs are the essential clinical drugs for colorectal cancer (CRC) treatment. However, the drug resistance of 5-FU has caused high mortality of CRC patients. Thus, it is urgent to develop reversal agents of 5-FU resistance. Sphingosine-1-phosphate receptor 2 (S1PR2) was proved to be a potential target for reversing 5-FU resistance, but the activity of known S1PR2 antagonists JTE-013 were weak in 5-FU-resistant cell lines. To develop more potent S1PR2 antagonists to treat 5-FU-resistant cancer, a series of JTE-013 derivatives were designed and synthesized. The most promising compound 40 could markedly reverse the resistance in 5-FU-resistant HCT116 cells and 5-FU-resistant SW620 cells via inhibiting the expression of dihydropyrimidine dehydrogenase (DPD). The key was that compound 40 with improved pharmacokinetic properties significantly increased the inhibitory rate of 5-FU in the SW620/5-FU cells xenograft model with no observable toxicity by inhibiting the expression of DPD in tumor and liver tissues. Altogether, these results suggest that compound 40 may be a promising drug candidate to reverse 5-FU resistance in the treatment of CRC.

Background. Cardiac fibrosis (CF) is major myocardial change in diabetic cardiomyopathy (DCM). Yangxinshi as a Chinese medicine formula is used to treat cardiovascular diseases. However, the exact effective mechanism of Yangxinshi on CF is still uncertain. Hence, based on the pharmacological network, predicting the active components, potential targets and pathways of Yangxinshi on diabetic fibrosis require to be further studied. Materials and Methods. By using Cytoscape 3.6.0 Bisogenet plug-in, the active components of Yangxinshi were obtained and screened through TCMSP, and the PPI network of DCM-CF was constructed and then screened by CytoNCA plug-in. GO analysis and KEGG pathway enrichment analysis were carried out by Cluego plug-in. Combined with the results of network pharmacological analysis, cells in vitro were performed to verify the CF stimulated with high glucose or intervence with Yangxinshi, and the expressions of Cbl-b, p-smad2, and α-SMA were detected. Results. Yangxinshi might play a key role in reversing cardiac fibrosis in individuals with DCM by regulating the signal pathway of CBL and promoted the expression of Cbl-b and inhibited the expression of p-smad2 and α-SMA, verifying some predictive work via network pharmacology. Conclusion. Based on network pharmacology, this study demonstrates that the beneficial effect of Yangxinshi on CF is related to the Cbl-b/smad2 pathway, providing an idea for the therapeutic effect of Yangxinshi on cardiac fibrosis in DCM.

Background: Sesamin is a rich phytochemical found in sesame seed oil that can promote osteoblast differentiation of rat BMSCs and improve rat bone structure by regulating Wnt/-Catenin pathway. Combined sesamin and γ-Tocotrienol (γ-T3) have been clarified to inhibit the proliferation of breast cancer cells, but their role in osteoporosis has not been explored. This paper aimed to discuss the synergistic effect of sesamin and γ-T3 in osteoporosis and disclose the underlying mechanism. Materials and methods: CCK-8 assay was to appraise the proliferation of hBMSCs after treated with sesamin and γ-T3. Moreover, the proteins in AMPK signaling in osteoblasts pretreated with AMPK inhibitor compound C (CC) were detected after the induction of sesamin and γ-T3. Then, CCK-8, ALP assay and ARS staining were used to analyze whether the proliferation and osteoblast differentiation of hBMSCs was via APMK pathway. RT-qPCR and western blot were conducted to quantify the levels of markers in osteoblasts. Results: It was determined that 5 g/mL sesamin and 1 μM γ-T3 exerted obvious influences on the viability of hBMSCs. Moreover, the co-treatment of sesamin and γ-T3 elevated the protein levels of related factors in AMPK pathway, which was reversed by CC. Furthermore, The proliferation and osteoblast differentiation exhibited remarkable increments upon exposure to both sesamin and γ-T3, whereas CC abolished these effects. Conclusion: In conclusion, the present study presented the first line of evidence to verify the synergystic effects of sesamin and γ-T3 on alleviating osteoporosis, and revealed their effects were realized by modulating the AMPK pathway. This paper has indicated the great potential of combined sesamin and γ-T3 in osteoporosis treatment.

This study aims to introduce a morphological classification of hyperextension tibial plateau fractures based on CT scans and to reveal the correlation between the anterior compression and posterior tension fractures. From January 2015 to January 2019, 37 patients with hyperextension tibial plateau fractures were studied retrospectively. Based on this classification, the fractures were divided into 2 groups: group A had anterolateral or anteromedial compression fractures while group B had both. Three observers classified the fractures and recorded the morphology and incidences of posterior plateau fractures and proximal fibular fractures. All 37 fractures were allocated to group A (n = 15; 40%) and B (n = 22; 60%). Of the posterior tibial plateau fractures, 10 (27%) fractures were defined as partial and 27 (73%) as total. Of the 37 fractures, 18 (49%) proximal fibular avulsion fractures were observed. There was a significant difference between groups A and B regarding the incidence of total posterior tibial plateau fractures (P .05). Hyperextension tibial plateau fractures with a decreased posterior slope angle always involve both the anteromedial and anterolateral plateaus. This CT-based classification may improve the understanding of fracture features and is helpful for planning treatment.

Allicin (2-propene-1-sulfinothioic acid S-2-propenyl ester), with quite a good range of hepatoprotective and antineoplastic properties, is a functional substance from garlic (Allium sativum L.) The purpose of this study was to provide evidence that allicin could protect 1,3-DCP-induced lipid metabolism disorder in HepG2 cells. Allicin reduced the accumulation of triglycerides (TG) and total cholesterol (TC) in 1,3-DCP-induced HepG2 cells. Allicin significantly increased the phosphorylation of AMP-activated protein kinase (AMPK) and down-regulated the levels of sterol regulatory element binding protein-1 (SREBP-1) and sterol regulatory element binding protein-2 (SREBP-2) in 1,3-DCP-induced HepG2 cells. Additionally, allicin had obvious recovery influence on the phosphorylation level of PKA and CREB in 1,3-DCP-induced HepG2 cells. These observations indicated that allicin alleviated lipid metabolism disorder induced by 1,3-DCP in HepG2 cells by regulating AMPK-SREBPs and PKA-CREB signaling pathways.

The TREX complex mediates the passage of bulk cellular mRNA export to the nuclear export factor TAP/NXF1 via the export adaptors ALYREF or UIF, which appear to act in a redundant manner. TREX complex recruitment to nascent RNA is coupled with 5′ capping, splicing and polyadenylation. Therefore to facilitate expression from their intronless genes, herpes viruses have evolved a mechanism to circumvent these cellular controls. Central to this process is a protein from the conserved ICP27 family, which binds viral transcripts and cellular TREX complex components including ALYREF. Here we have identified a novel interaction between HSV-1 ICP27 and an N-terminal domain of UIF in vivo, and used NMR spectroscopy to locate the UIF binding site within an intrinsically disordered region of ICP27. We also characterized the interaction sites of the ICP27 homolog ORF57 from KSHV with UIF and ALYREF using NMR, revealing previously unidentified binding motifs. In both ORF57 and ICP27 the interaction sites for ALYREF and UIF partially overlap, suggestive of mutually exclusive binding. The data provide a map of the binding sites responsible for promoting herpes virus mRNA export, enabling future studies to accurately probe these interactions and reveal the functional consequences for UIF and ALYREF redundancy.

We study the terahertz transmission properties in mirrored asymmetric split ring metamaterials which were fabricated by using the method of microstructures compensation. Combined with the analysis of finite difference time domain, the distribution of terahertz electric field and current density are discussed in metamaterials structures. Our experiments show that the quality factor in compensated microstructures is 12.5 at the frequency of 0.92 THz which is higher than that of 8.29 in the uncompensated microstructures. The estimation of THz energy stored in the capacitor configuration indicate that the stored energy in compensated structures is 1.46 times that of in uncompensated structures, which resulted in the enhancement of quality factor about 50% in the compensated microstructures.

OBJECTIVE This study aims to observe the efficacies of microsurgical technique combined with the Zhuang's skin soft tissue expander in treating Hunter type III congenital microtia. METHODS Fifty-eight patients (61 ears) were enrolled from 2003 to 2012; the skin tissue expander was embedded subepidermally in the first stage via the intrahairline longitudinal incision in the postauricular mastoid area, the diseased-side rib cartilage was then taken for preparing the ear bracket in the second stage, and the tragus was surgically reconstructed in the third stage. RESULTS The mean follow-up lasted 6 months to 10 years, the results were satisfactory for 54 ears and acceptable for 5 ears, and 2 ears (in two patients) appeared with complications, including 1 case of ear flap expansion rupture and 1 case of postoperative lateral helix flap necrosis. CONCLUSIONS The combination of microsurgical technique, Zhuang's skin soft tissue expander, and autogenous rib cartilage graft could achieve satisfactory results in treating Hunter type III congenital microtia, and the complications were less, so it was worthy of clinical applications.

In hepatitis B virus (HBV) replication cycle, pregenomic RNA undergoes splicing and the reverse transcribed defective genomes can be packaged and released. Various types of spliced defective HBV genomes have been isolated from the sera and liver tissues of viral hepatitis B patients. To explore the functions of a 2.2 kb double spliced HBV variant, a 3.2 kb full-length HBV isolate (#97–34) and its 2.2 kb double-spliced HBV variant (#AP–12) from the tumor tissue of a patient with hepatocellular carcinoma (HCC) were amplified and cloned. Sequencing results showed that #AP12 had deletions in pre-S2, part of pre-S1, S genes, part of the spacer, and part of the reverse transcriptase gene, while the X gene was intact. When this defective double-spliced genome and its full-length counterpart genome were co-transfected into HepG2 cells, the former was shown to enhance the replication of the latter, both by real-time PCR and Southern blotting. When a replication incompetent clone 97–34G1881A was used to co-transfect with #AP12, #AP12 DNA was increased, indicating that replication of the wild-type virus was not the only factor involved in this observation. However, the replication enhancing competency of #AP12 was shown to require an intact HBV X expression cassette. The double-spliced defective variant might contribute to persistent HBV replication in a subpopulation of HCC patients. J. Med. Virol. 81:230–237, 2009. © 2008 Wiley-Liss, Inc.

Background PR interval variations have recently been associated with an increased risk of long-term atrial fibrillation (AF), heart block and all-cause mortality. Genome-wide association studies have linked the PR interval with several common variants in the TBX5 gene. Several variants in the TBX5 gene, including rs7312625 and rs883079, have been associated with AF. The purpose of this study was to determine the association of single-nucleotide polymorphisms (SNPs) in the TBX5 gene, rs7312625 and rs883079, with AF in Chinese Han patients. Methodology/Principal Findings In this case-control association study, large cohorts of AF patients (n = 1132) and controls (n = 1206) were recruited from different hospitals. The genotyping was performed using a Rotor-Gene TM 6000 high-resolution melt system. Rs7312625, rs3825214 and rs883079 were analyzed. We found that SNP 3825214 was significantly associated with AF (P-obs = 0.002, odds ratio [OR] = 0.82), and lone AF (P-obs = 6.77x10-5, odds ratio [OR] = 0.71). SNP rs7312625 was significantly associated with lone AF (P-obs = 0.015, odds ratio [OR] = 1.27), although its association with AF was not significant. No significant association of SNP rs883079 with AF or lone AF was observed. Thus, we analyzed the interaction among these three loci. We demonstrated significant interaction among rs3825214, rs7312625 and rs883079. Four-locus risk alleles showed the highest odds ratio in combined rs3825214 and rs7312625 (P-obs

Circulating tumor cells (CTCs) are becoming promising biomarkers in several cancers, such as colon, prostate, and breast carcinomas. Independent research groups have reported a correlation between CTC numbers and patient prognosis. Even more, the development of personalized medicine gives physicians impetus to utilize the advancement of molecular characterization of CTCs. This review introduces a new technique, CTCscope, and compares it with the current methods of CTCs detection, with particular emphasis on cancer research, and discusses the future application of this new method from bench to bed-side.

Herpes simplex virus 1 (HSV-1) protein ICP27 enables viral mRNA export by accessing the cellular mRNA export receptor TAP/NXF, which guides mRNA through the nuclear pore complex. ICP27 binds viral mRNAs and interacts with TAP/NXF, providing a link to the cellular mRNA export pathway. ICP27 also interacts with the mRNA export adaptor protein Aly/REF, which binds cellular mRNAs and also interacts with TAP/NXF. Studies using small interfering RNA (siRNA) knockdown indicated that Aly/REF is not required for cellular mRNA export, and similar knockdown studies during HSV-1 infection led us to conclude that Aly/REF may be dispensable for viral RNA export. Recently, the structural basis of the interaction of ICP27 with Aly/REF was elucidated at atomic resolution, and it was shown that three ICP27 residues, W105, R107, and L108, interface with the RNA recognition motif (RRM) domain of Aly/REF. Here, to determine the role the interaction of ICP27 and Aly/REF plays during infection, these residues were mutated to alanine, and a recombinant virus, WRL-A, was constructed. Virus production was reduced about 10-fold during WRL-A infection, and export of ICP27 protein and most viral mRNAs was less efficient. We conclude that interaction of ICP27 with Aly/REF contributes to efficient viral mRNA export.

Cyclophilin A (CypA), predominantly located intracellularly, is a multifunctional protein. We previously reported decreased CypA levels in hepatocytes of transgenic mice expressing hepatitis B virus (HBV) surface antigen (HBsAg). In this study, we found that expression of HBV small surface protein (SHBs) in human hepatoma cell lines specifically triggered CypA secretion, whereas SHBs added extracellularly to culture medium did not. Moreover, CypA secretion was not promoted by the expression of a secretion deficient SHBs mutant, suggesting a close association between secretion of CypA and SHBs. Interaction between CypA and SHBs was observed by using coimmunoprecipitation and glutathione S -transferase pull-down assays. Hydrodynamic injection of the SHBs expression construct into C57BL/6J mice resulted in increased serum CypA levels and ALT/AST levels, as well as the infiltration of inflammatory cells surrounding SHBs-positive hepatocytes. The inflammatory response and serum ALT/AST level were reduced when the chemotactic effect of CypA was inhibited by cyclosporine and anti-CD147 antibody. Furthermore, higher serum CypA levels were detected in chronic hepatitis B patients than in healthy individuals. In HBV patients who had received liver transplantation, serum CypA levels declined dramatically after the loss of HBsAg as a consequence of liver transplantation. Taken together, these results indicate that expression and secretion of SHBs can promote CypA secretion, which may contribute to the pathogenesis of HBV infection.

Accumulating evidence suggests a key role of hepatocyte apoptosis in the pathogenesis of viral hepatitis B. It was found in this study that stable expression of small hepatitis B surface antigen (SHBs) in HepG2 and Huh7 cells increased susceptibility to apoptosis. Proteomic analysis of SHBs expressing HepG2 cells revealed 43 down-regulated and 38 up-regulated proteins. Some have been implicated in apoptosis, including glucose-regulated protein 78 kDa (GRP78), heterogeneous nuclear ribonucleoprotein H3 (hnRNP H), Rho GDP dissociation inhibitor (GDI), cystatin B, far upstream element-binding protein (FUSEbp), and TNF receptor-associated protein 1 (TRAP1). Differential expression of GRP78 and several other proteins was confirmed by Western blot analysis. Replenishing GRP78 improved cellular resistance to apoptosis, whereas reduction of GRP78 by siRNA increased susceptibility even in the absence of SHBs. Taken together, these results suggest that HBsAg plays a pro-apoptotic role through down-regulation of GRP78.

The genome of hepatitis B virus (HBV) consists of four open reading frames, encoding the envelope proteins (Pre-S/S), the core proteins (Pre-C/C), the polymerase (P) and the transactivating X protein (X). In the sera of HBV-infected patients, hepatitis B surface antigen (HBsAg) particles without the viral genome can outnumber virions by more than 1000-fold. To analyse the interactions between HBsAg and host cells, global gene-expression profiles of a small HBsAg (SHBs)-secreting stable cell line (HepG2-S-G2) and its counterpart control cell line (HepG2-Neo-F4) were compared. Marked upregulation of lymphoid enhancer-binding factor 1 (LEF-1), a transcription factor in the Wnt pathway, was found in SHBs-expressing cells and was confirmed by interference experiments with small interfering RNA. However, compared with the control cells, HepG2-S-G2 did not show higher proliferative competence in culture or increased tumorigenesis in nude mice. A possible mechanism to explain the discrepancy between the upregulation of LEF-1 and the lack of increased tumorigenesis is SHBs expression resulting in altered expression and distribution of LEF-1 protein in cell compartments and upregulation of LEF-1 isoforms that could suppress, rather than enhance, the Wnt pathway.

The small, 22-nm spherical particles associated with hepatitis B infection are composed of hepatitis B surface antigen (HBsAg) and usually outnumber the virions by a ratio of 10(2) or 10(3). To study the interactions and pathogenesis between liver cells and the expression of HBsAg, global protein profiles were compared by two dimensional gel-based differential proteomics between the livers of a lineage of HBsAg positive transgenic mice and their HBsAg negative control siblings. A total of 93 proteins were identified in the HBV transgenic mice. Around 45% of these differentially expressed proteins were enzymes associated with metabolism, suggesting that the processing of lipids, carbohydrates and certain amino acids were up- or down-regulated. Among these proteins, cyclophilin A (CypA), the major target for the potent immunosuppressive drug cyclosporin A, was found decreased in HBsAg positive transgenic mouse liver and in a stable cell line expressing HBsAg when compared to their controls. The decrease of intracellular CypA was accompanied by an increased secretion of this protein into the supernatant of HBsAg positive cells. Possible implications of HBsAg expression and the intracellular decrease of CypA are discussed.

To study effective method of showing the microvascular architecture of human and rabbit skin flaps.According to endogenous alkaline phosphatase activity, Gomori-Takamatsu method and NBT/BCIP method were used to observe the microvascular architecture.Both methods had capability to demonstrate the microvascular architecture of the human skin flap, whereas the examining results of rabbits were negative.Great difference of endogeneous alkaline phosphatase activity exists in different animals. Gomori-Takamatsu method and NBT/BCIP method are useful in study of microvascular architecture of human skin flaps.

Background A prolonged PR interval is a sign of increased risk of cardiac arrhythmia. Recent genome-wide association studies found that the single-nucleotide polymorphism (SNP) rs3825214 in T-box 5 (TBX5) was positively associated with PR interval, QRS duration, QT interval, and common arrhythmia disorders such as atrial fibrillation (AF) and advanced atrioventricular block. However, other independent replication studies are required to validate the result. This study assessed associations between rs3825214 and ECG parameters, AF, and ventricular tachycardia (VT) in a Chinese Han population. Methodology/Principal Findings To assess the association between rs3825214 and AF and VT, we carried out case-control association studies with 692 AF patients (including 275 lone AF patients), 235 VT patients, and 856 controls. Genotyping was performed using a Rotor-Gene TM 6000 High Resolution Melt system. Statistical analyses of associations were adjusted for potential confounding factors. A moderate association was detected between rs3825214 and AF (Padj = 0.036, OR = 0.79) and a highly significant association was detected between the G allele of rs3825214 and lone AF (Padj = 0.001, OR = 0.65; genotypic P = 3.75×10−4 with a dominant model). We also found that rs3825214 showed a significant association with atrial-ventricular block (AVB; P = 0.028; Padj = 0.035, OR = 0.494). Conclusions Our results indicate that rs3825214 conferred a significant risk of lone AF in this Chinese Han population.

Background There are around 350 million of hepatitis B surface antigen (HBsAg) carriers worldwide, and among them, high risk of developing hepatocellular carcinoma (HCC) has been identified by epidemiological studies. To date, the molecular role of HBsAg in HCC development has not been fully studied. We have previously reported that in cell cultures, HBsAg up-regulated the expression of lymphoid enhancer-binding factor 1 (LEF-1), a key component of the Wnt pathway. In this study we aimed to study this effect of HBsAg on LEF-1 in the development of HCC. Methods Expression of HBsAg, LEF-1 and its downstream effector genes were compared among 30 HCCs, their peritumor tissue counterparts and 9 normal control liver tissues by quantitative real-time PCR. In addition, immunohistochemical staining studies on HBsAg and LEF-1 expression were conducted among these samples. Results The expression of LEF-1 was compared between 13 HBsAg positive HCC tissues and 17 HBsAg negative HCC tissues. Simultaneous detection of LEF-1 and HBsAg was observed in HBsAg positive HCC tissues and, additionally, the simultaneous detection of HBsAg and LEF-1 was more pronounced in peritumor tissues, compared to that in the tumor tissues. The distribution of cellular LEF-1 in peritumor tissues was predominantly in the cytoplasm; while LEF-1 in the tumor tissues was located either exclusively in the nucleus or both in the nucleus and cytoplasm. By real-time PCR, the expression levels of LEF-1 downstream effector genes cyclin D1 and c-myc were higher in peritumor cells compared to that of the tumor cells. However, a 38 kDa truncated isoform of LEF-1, rather than the 55 kDa wild-type LEF-1, was significantly elevated in the HBsAg positive tumor cells. Conclusion Data indicate that deregulation of the Wnt pathway by HBsAg occurred in HBV-associated HCCs, but was more pronounced in the peritumor cells. It is speculated that HBsAg could stimulate proliferation and functional modification of hepatocytes via LEF-1 through the Wnt pathway at the pre-malignant stage.