Non-syntrophic methanogenic hydrocarbon degradation by an archaeal species.

The methanogenic degradation of oil hydrocarbons can proceed through syntrophic partnerships of hydrocarbon-degrading bacteria and methanogenic archaea1-3. However, recent culture-independent studies have suggested that the archaeon 'Candidatus Methanoliparum' alone can combine the degradation of long-chain alkanes with methanogenesis4,5. Here we cultured Ca. Methanoliparum from a subsurface oil reservoir. Molecular analyses revealed that Ca. Methanoliparum contains and overexpresses genes encoding alkyl-coenzyme M reductases and methyl-coenzyme M reductases, the marker genes for archaeal multicarbon alkane and methane metabolism. Incubation experiments with different substrates and mass spectrometric detection of coenzyme-M-bound intermediates confirm that Ca. Methanoliparum thrives not only on a variety of long-chain alkanes, but also on n-alkylcyclohexanes and n-alkylbenzenes with long n-alkyl (C≥13) moieties. By contrast, short-chain alkanes (such as ethane to octane) or aromatics with short alkyl chains (C≤12) were not consumed. The wide distribution of Ca. Methanoliparum4-6 in oil-rich environments indicates that this alkylotrophic methanogen may have a crucial role in the transformation of hydrocarbons into methane.

Globular adiponectin induces esophageal adenocarcinoma cell pyroptosis via the miR-378a-3p/UHRF1 axis.

BACKGROUND: Antiapoptosis is a major factor in the resistance of tumor cells to chemotherapy and radiotherapy. Thus, activation of cell pyroptosis may be an effective option to deal with antiapoptotic cancers such as esophageal adenocarcinoma (EAC). METHODS: Differential expression of ubiquitin-like versus PHD and ring finger structural domain 1 (UHRF1) in EAC and near normal tissues was analyzed, as well as the prognostic impact on survival in EAC. Also, the same study was done for globular adiponectin (gAD). Simultaneously, the mRNA expression of UHRF1 was observed in different EAC cell lines. Real time cellular analysis (RTCA) was used to detect cell proliferation, and flow cytometry and inverted fluorescence microscopy were used to detect pyroptosis. Biocredit analysis was conducted to observe the correlation between UHRF1 and key pyroptosis proteins. OD values and CCK8 assay were used to determine the effect of miR-378a-3p on EAC cells. Quantitative real-time polymerase chain reaction and Western blot were used to detect the correlation between UHRF1, gAD, and miR-378a-3p in EAC cells. Moreover, in vivo and in vitro experiments were performed to detect the relevant effects on tumor migration and invasion after inhibiting UHRF1 expression. RESULTS: UHRF1 was negatively correlated with the survival of patients with EAC, while miR-378a-3p showed the opposite effect. Additionally, gAD promoted EAC cell pyroptosis, upregulated miR-378a-3p, and significantly inhibited the proliferation of EAC cells. gAD directly reduced UHRF1 expression in EAC cells by upregulating miR-378a-3p. In cell migration and invasion assays, inhibition of UHRF1 expression significantly suppressed EAC cell metastasis. In animal experiments, we again demonstrated that gAD induced pyroptosis in EAC cells by inhibiting the expression of UHRF1. CONCLUSION: gAD-induced upregulation of miR-378a-3p significantly inhibited the proliferation of EAC by targeting UHRF1. Therefore, gAD may serve as an alternative therapy for chemotherapy- and radiation-refractory EAC or other cancers with the same mechanism of pyroptosis action.

Illuminating the biosynthesis pathway genes involved in bioactive specific monoterpene glycosides in Paeonia veitchii Lynch by a combination of sequencing platforms.

BACKGROUND: Paeonia veitchii Lynch, a well-known herb from the Qinghai-Tibet Plateau south of the Himalayas, can synthesize specific monoterpene glycosides (PMGs) with multiple pharmacological activities, and its rhizome has become an indispensable ingredient in many clinical drugs. However, little is known about the molecular background of P. veitchii, especially the genes involved in the biosynthetic pathway of PMGs. RESULTS: A corrective full-length transcriptome with 30,827 unigenes was generated by combining next-generation sequencing (NGS) and single-molecule real-time sequencing (SMRT) of six tissues (leaf, stem, petal, ovary, phloem and xylem). The enzymes terpene synthase (TPS), cytochrome P450 (CYP), UDP-glycosyltransferase (UGT), and BAHD acyltransferase, which participate in the biosynthesis of PMGs, were systematically characterized, and their functions related to PMG biosynthesis were analysed. With further insight into TPSs, CYPs, UGTs and BAHDs involved in PMG biosynthesis, the weighted gene coexpression network analysis (WGCNA) method was used to identify the relationships between these genes and PMGs. Finally, 8 TPSs, 22 CYPs, 7 UGTs, and 2 BAHD genes were obtained, and these putative genes were very likely to be involved in the biosynthesis of PMGs. In addition, the expression patterns of the putative genes and the accumulation of PMGs in tissues suggested that all tissues are capable of biosynthesizing PMGs and that aerial plant parts could also be used to extract PMGs. CONCLUSION: We generated a large-scale transcriptome database across the major tissues in P. veitchii, providing valuable support for further research investigating P. veitchii and understanding the genetic information of plants from the Qinghai-Tibet Plateau. TPSs, CYPs, UGTs and BAHDs further contribute to a better understanding of the biology and complexity of PMGs in P. veitchii. Our study will help reveal the mechanisms underlying the biosynthesis pathway of these specific monoterpene glycosides and aid in the comprehensive utilization of this multifunctional plant.

Comparison of American and European guidelines for cardio-oncology of heart failure.

Heart failure is a complex clinical syndrome, whose signs and symptoms are caused by functional or structural impairment of ventricular filling or ejection of blood. Due to the interaction among anticancer treatment, patients' cardiovascular background, including coexisting cardiovascular diseases and risk factors, and cancer itself, cancer patients develop heart failure. Some drugs for cancer treatment may cause heart failure directly through cardiotoxicity or indirectly through other mechanisms. Heart failure in turn may make patients lose effective anticancer treatment, thus affecting the prognosis of cancer. Some epidemiological and experimental evidence shows that there is a further interaction between cancer and heart failure. Here, we compared the cardio-oncology recommendations among heart failure patients of the recent 2022 American guidelines, 2021 European guidelines, and 2022 European guidelines. Each guideline acknowledges the role of multidisciplinary (cardio-oncology) discussion before and during scheduled anticancer therapy.

Multidimensional analysis to elucidate the possible mechanism of bone metastasis in breast cancer.

BACKGROUND: Breast cancer (BC) patients tend to suffer from distant metastasis, especially bone metastasis. METHODS: All the analysis based on open-accessed data was performed in R software, dependent on multiple algorithms and packages. The RNA levels of specific genes were detected using quantitative Real-time PCR as a method of detecting the RNA levels. To assess the ability of BC cells to proliferate, we utilized the CCK8 test, colony formation, and the 5-Ethynyl-20-deoxyuridine assay. BC cells were evaluated for invasion and migration by using Transwell assays and wound healing assays. RESULTS: In our study, we identified the molecules involved in BC bone metastasis based on the data from multiple BC cohorts. Then, we comprehensively investigated the effect pattern and underlying biological role of these molecules. We found that in the identified molecules, the EMP1, ACKR3, ITGA10, MMP13, COL11A1, and THY1 were significantly correlated with patient prognosis and mainly expressed in CAFs. Therefore, we explored the CAFs in the BC microenvironment. Results showed that CAFs could activate multiple carcinogenic pathways and most of these pathways play an important role in cancer metastasis. Meanwhile, we noticed the interaction between CAFs and malignant, endothelial, and M2 macrophage cells. Moreover, we found that CAFs could induce the remodeling of the BC microenvironment and promote the malignant behavior of BC cells. Then, we identified MMP13 for further analysis. It was found that MMP13 can enhance the malignant phenotype of BC cells. Meanwhile, biological enrichment and immune infiltration analysis were conducted to present the effect pattern of MMP13 in BC. CONCLUSIONS: Our result can improve the understanding of researchers on the underlying mechanisms of BC bone metastasis.

DEGENERATED LEMMA (DEL) regulates lemma development and affects rice grain yield.

In grass, the lemma is a unique floral organ structure that directly determines grain size and yield. Despite a great deal of research on grain enlargement caused by changes in glume cells, the importance of normal development of the glume for normal grain development has been poorly studied. In this study, we investigated a rice spikelet mutant, degenerated lemma (del), which developed florets with a slightly degenerated or rod-like lemma. More importantly, del also showed a significant reduction in grain length and width, seed setting rate, and 1000-grain weight, which led to a reduction in yield. The results indicate that the mutation of the DEL gene further affects rice grain yield. Map-based cloning shows a single-nucleotide substitution from T to A within Os01g0527600/DEL/OsRDR6, causing an amino acid mutation of Leu-34 to His-34 in the del mutant. Compared with the wild type, the expression of DEL in del was significantly reduced, which might be caused by single base substitution. In addition, the expression level of tasiR-ARF in del was lower than that of the wild type. RT-qPCR results show that the expression of some floral organ identity genes was changed, which indicates that the DEL gene regulates lemma development by modulating the expression of these genes. The present results suggest that the normal expression of DEL is necessary for the formation of lemma and the normal development of grain morphology and therefore has an important effect on the yield. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01297-6.

[Selection and validation of reference genes for quantitative real-time PCR analysis in Paeonia veitchii].

Paeonia veitchii and P. lactiflora are both original plants of the famous Chinese medicinal drug Paeoniae Radix Rubra in the Chinese Pharmacopoeia. They have important medicinal value and great potential in the flower market. The selection of stable and reliable reference genes is a necessary prerequisite for molecular research on P. veitchii. In this study, two reference genes, Actin and GAPDH, were selected as candidate genes from the transcriptome data of P. veitchii. The expression levels of the two candidate genes in different tissues(phloem, xylem, stem, leaf, petiole, and ovary) and different growth stages(bud stage, flowering stage, and dormant stage) of P. veitchii were detected using real-time fluorescence quantitative technology(qRT-PCR). Then, the stability of the expression of the two reference genes was comprehensively analyzed using geNorm, NormFinder, BestKeeper, ΔCT, and RefFinder. The results showed that the expression patterns of Actin and GAPDH were stable in different tissues and growth stages of P. veitchii. Furthermore, the expression levels of eight genes(Pv-TPS01, Pv-TPS02, Pv-CYP01, Pv-CYP02, Pv-CYP03, Pv-BAHD01, Pv-UGT01, and Pv-UGT02) in different tissues were further detected based on the transcriptome data of P. veitchii. The results showed that when Actin and GAPDH were used as reference genes, the expression trends of the eight genes in different tissues of P. veitchii were consistent, validating the reliability of Actin and GAPDH as reference genes for P. veitchii. In conclusion, this study finds that Actin and GAPDH can be used as reference genes for studying gene expression levels in different tissues and growth stages of P. veitchii.

MYBL2 regulates de novo purine synthesis by transcriptionally activating IMPDH1 in hepatocellular carcinoma cells.

BACKGROUND: Metabolic reprogramming is a hallmark of cancer, alteration of nucleotide metabolism of hepatocellular carcinoma (HCC) is not well-understood. MYBL2 regulates cell cycle progression and hepatocarcinogenesis, its role in metabolic regulation remains elusive. PATIENTS AND METHODS: Copy number, mRNA and protein level of MYBL2 and IMPDH1 were analyzed in HCC, and correlated with patient survival. Chromatin Immunoprecipitation sequencing (Chip-seq) and Chromatin Immunoprecipitation quantitative polymerase chain reaction (ChIP-qPCR) were used to explore the relationship between MYBL2 and IMPDH1. Metabolomics were used to analyze how MYBL2 affected purine metabolism. The regulating effect of MYBL2 in HCC was further validated in vivo using xenograft models. RESULTS: The Results showed that copy-number alterations of MYBL2 occur in about 10% of human HCC. Expression of MYBL2, IMPDH1, or combination of both were significantly upregulated and associated with poor prognosis in HCC. Correlation, ChIP-seq and ChIP-qPCR analysis revealed that MYBL2 activates transcription of IMPDH1, while knock-out of MYBL2 retarded IMPDH1 expression and inhibited proliferation of HCC cells. Metabolomic analysis post knocking-out of MYBL2 demonstrated that it was essential in de novo purine synthesis, especially guanine nucleotides. In vivo analysis using xenograft tumors also revealed MYBL2 regulated purine synthesis by regulating IMPDH1, and thus, influencing tumor progression. CONCLUSION: MYBL2 is a key regulator of purine synthesis and promotes HCC progression by transcriptionally activating IMPDH1, it could be a potential candidate for targeted therapy for HCC.

The comparison between the effect of Glycyrrhizae uralensis (Gan-Cao) and Montelukast on the expression of T-bet and GATA-3 genes in children with allergic asthma.

Pediatric allergic asthma is a chronic disease that affects the lungs and airways. If a child is exposed to certain stimulants such as pollen inhalation, colds, or respiratory infections, the lungs become inflamed and if left untreated can lead to dangerous asthma attacks. One of the most important treatments for this disease is the use of leukotriene modulators, such as montelukast. But recently, due to easier access, cheaper prices and fewer side effects, attention has shifted to non-chemical treatments. Gan-Cao (Glycyrrhizae uralensis), as traditional Chinese medicine, has been proved to have a good therapeutic effect on experimental allergic asthma. But its anti-asthma mechanism is currently unclear. Therefore, the study aimed the comparison between the effect of Gan-Cao and montelukast on the expression of T-bet and GATA-3 genes in children with allergic asthma. For this purpose, fifty children with allergic asthma were divided into two groups. The first group was treated with montelukast for one month. The second group was treated with Gan-Cao root extract. Then the peripheral blood mononuclear cells were isolated, their RNA was extracted, and the relative expression of T-bet and GATA3 transcription factors was evaluated by Real-time PCR. The relationship between them and risk factors for asthma was assessed by relevant statistical tests. The result showed the expression of the GATA3 gene (P = 0.102), T-bet gene (P = 0.888), and the expression ratio of T-bet/GATA-3 genes (P = 0.061) was not significantly different between the two groups. It showed that Gan-Cao can affect the expression of these genes just as much as montelukast. Therefore, this Chinese herb can be used as an alternative or supplement medicine to treat allergic asthma in children.

Morphological and Age-Related Changes in the Narcolepsy Brain.

Morphological changes in the cortex of narcolepsy patients were investigated by surface-based morphometry analysis in this study. Fifty-one type 1 narcolepsy patients and 60 demographically group-matched healthy controls provided resting-state functional and high-resolution 3T anatomical magnetic resonance imaging scans. Vertex-level cortical thickness (CT), gyrification, and voxel-wise functional connectivity were calculated. Adolescent narcolepsy patients showed decreased CT in bilateral frontal cortex and left precuneus. Adolescent narcolepsy demonstrated increased gyrification in left occipital lobe, left precuneus, and right fusiform but decreased gyrification in left postcentral gyrus, whereas adult narcolepsy exhibited increased gyrification in left temporal lobe and right frontal cortex. Furthermore, sleepiness severity was associated with altered CT and gyrification. Increased gyrification was associated with reduced long-range functional connectivity. In adolescent patients, those with more severe sleepiness showed increased right postcentral gyrification. Decreased frontal and occipital gyrification was found in cases with hallucination. In adult patients, a wide range of regions showed reduced gyrification in those with adolescence-onset compared adult-onset narcolepsy patients. Particularly the frontal lobes showed altered brain morphology, being a thinner cortex and more gyri. The impact of narcolepsy on age-related brain morphological changes may remain from adolescence to young adulthood, and it was especially exacerbated in adolescence.

First Report of Chinese Cabbage Wilt Caused by Plectosphaerella cucumerina in Inner Mongolia of China.

Chinese cabbage [Brassica rapa L. subsp. pekinensis (Lour.) Hanelt] has been grown commercially for many decades in Huade County, Inner Mongolia. In 2018 and 2019, an unusual stem and leaf wilt disease with an average disease incidence of approximately 3% was observed. Diseased plants with spindle-shaped stem lesions were collected and small pieces (0.3 × 0.3 cm) of the diseased tissues were cut from the margins of stem lesions, surface sterilized with 75% alcohol for 3 to 5 s, 0.1% NaClO for 2 to 3 min, and washed three times with sterilized water. The treated tissues were placed on 1.5% (w/v) water agar plates and incubated at 25°C for 3 days. The mycelia were cut and transferred onto potato dextrose agar (PDA) for culture purification. Three isolates with similar morphology were obtained and named as BC-2, BC3-2 and BG2. To confirm their pathogenicity, Chinese cabbage (cv. Chunqiuhuang) seed was planted into plugs. After 30 days, the fibrous roots were wounded with a fruit knife and root-dipped in the conidium suspension (1 × 106 conidia/ml) for 20 min. Inoculated seedlings were transplanted in pots (30 × 25 cm) with sterilized nursery soil, with one seeding per pot. The roots of control plants were also wounded and dipped in sterilized water. Five seedlings were inoculated with each isolate and the experiment was repeated three times. Treated seedlings were maintained in a greenhouse at 25 to 28°C under a 12-h photoperiod. Chlorosis and wilting were observed approximately 4 weeks after inoculation, and the outer layer of leaves of the inoculated seedlings developed discoloration and wilted symptoms after 50 days. The symptoms induced by all three isolates were the same as the symptoms observed in the field, whereas no symptoms developed on the control plants. To confirm the Koch's postulates, the fungus was successfully re-isolated from the infected leaves and had similar growth and morphology as the original isolates. The three isolates were cultured for both morphology and molecular identification. The 14-day-old colonies on PDA were buff or salmon pink with few aerial hyphae, and slimy surfaces. Aerial hyphae were sparse with simple or branched conidiophores. Conidia were ellipsoidal, hyaline, surface smooth, septate or aseptate, and (4.0 to 9.7 µm × 2.0 to 3.9 µm). Such characteristics are typical of Plectosphaerella spp. (Palm et al. 1995). For molecular identification, the internal transcribed spacer (ITS) region of rDNA was amplified using the primer pair ITS1/ITS4 (White et al. 1990), and the products were directly sequenced. BLAST analysis showed that the sequences of Isolates BC-2 (511 bp out of 515 bp), BC3-2 (512 bp out of 516 bp) and BG2 (503 bp out of 505 bp) showed 99% identity to an isolate of P. cucumerina (acc. no. KT826571.1) from Bottle gourd [Lagenaria siceraria (Molina) Standl.] (Yan et al. 2016). The sequences of Isolates BC-2, BC3-2 and BG2 were deposited in GenBank (acc. nos. MW320463, MW320462 and MW320464). Although P. cucumerina was reported causing root rot of cabbage (B. oleracea) in Gansu, China (Li et al. 2017), to our knowledge, this is the first report of P. cucumerina causing Chinese cabbage wilt in Inner Mongolia, China. The presence of the disease could cause significant economic losses in Chinese cabbage production. For this reason, strategies for the management and control of this disease should be implemented.

[Research progress on chemical constituents,pharmacological activities,and quality control of Patrinia villosa].

Patrinia villosa, regarding its functions in clearing heat and detoxification and eliminating carbuncles and pus, is widely used as a traditional medicinal herb that contains rich nutrition and substances such as various amino acids, vitamins, and soluble su-gar, and it is also an edible wild herb in Chinese folk tradition for 2 000 years. In 1973, Japanese scholars firstly separated three iridoids from Japanese P. villosa, and by 2021, chemical components such as flavonoids, iridoids, organic acids, triterpenoids, phenylpropanoids, and steroids have been found, which have multiple pharmacological effects, including antioxidant, antitumor, anti-diarrhea, antibacterial, sedative, and liver protection capabilities. Studies indicate that flavonoids, saponins, phenylpropanoids, and triterpenoids in P. villosa are vital substances for its pharmacological activities. However, the quality of this medicinal material cannot be controlled due to the unclear records in ancient books in the past dynasties and different drug use habits in different places, and thus its circulation is chaotic. At present, researchers have used flavonoids, organic acids, phenylpropanoids, triterpenoid saponins, and other compounds to conduct studies in this regard. Therefore, on the basis of the existing literature resources, we comprehensively summarize the chemical constituents, pharmacological activities, and quality control of P. villosa to further provide a reference for the safety and effectiveness of clinical drug use and lay a foundation for the follow-up experimental research.

Research on a method of eliminating the line broadening caused by the wavelength shift based on the first harmonic.

Using laser spectroscopy for gas detection, short-term rapid changes in ambient temperature, system noise, and circuit aging are likely to cause line broadening, which affects the accuracy of gas concentration measurement. Firstly, the correction method of the impact on line broadening is analyzed theoretically. A method is proposed to eliminate the line broadening caused by the wavelength shift based on the first harmonic detection. After removing the background noise and filtering, the standard harmonic fitting and broadening elimination are carried out. Meanwhile, a methane gas detection system is established and experiments are conducted. The experimental results show that after the standard harmonic fitting, the maximum value of the baseline is reduced from 2 to 0.078, and the maximum absolute value of the baseline in the absorption-free region is reduced from 2.07 to 0.072. The standard deviation after the broadening correction is 0.047, and the standard deviation without considering the effect of broadening is 0.203, which proves that the accuracy of trace gas detection is improved and has good engineering practical value.

Ustekinumab trough concentration affects clinical and endoscopic outcomes in patients with refractory Crohn's disease: a Chinese real-world study.

BACKGROUND: Ustekinumab (UST), a newly-used biologic targeting p40 subunit of IL12 and IL23 in China, exerts a confirmed therapeutic effect on the induction and maintenance therapies for refractory Crohn's disease (CD). Therapeutic drug monitoring based on trough and antibody concentration is of core importance when treating patients who lose response to UST. We aimed to analyze the UST exposure-response relationship in CD treatment in the real-world setting. METHODS: We retrospectively enrolled patients with CD who received UST between March 1, 2020 and May 31, 2021, at the inflammatory bowel disease (IBD) center of the Sun Yat-Sun Affiliated Sixth Hospital. Baseline characteristic information, biomarker examination, clinical outcomes determined by the Crohn's disease activity index (CDAI), and endoscopic outcomes evaluated using a simple endoscopic score for Crohn's disease (SES-CD) at week 16/20 were collected. The optimal UST cut-off trough concentration was identified using receiver operating characteristic curve (ROC) analysis. RESULTS: Nineteen eligible patients were included in the study, the mean age was 29.1 ± 9.1 years and the mean disease duration was 5.5 ± 4.7 years. At the initiation of the study, 89.5% of the patients had been exposed to prior biologics, 42.1% had previous CD-related surgeries, and 52.6% had perianal diseases. At week 16/20 after the UST initiation, clinical response, clinical remission, endoscopic response, and endoscopic remission were 89.5%, 84.2%, 42.2%, and 73.7%, respectively. The cut-off optimal trough concentration for UST was 1.12 µg/mL, as determined by the ROC with an area under the curve (AUC) of 0.78, sensitivity of 87.5%, and specificity of 72.7%. Patients with a UST trough concentration > 1.12 µg/mL had a significantly higher rate of endoscopic remission than those without (70.0% vs. 11.1%, P = 0.02). CONCLUSIONS: UST is an effective therapeutic option for refractory CD treatment. A UST trough concentration above 1.12 µg/mL was associated with endoscopic remission at week 16/20 after UST initiation. Trial registration This study was approved and retrospectively registered by the Ethics Committee of Sun Yat-Sen University (2021ZSLYEC-066, March 29, 2021) and the Clinical Trial Registry (NCT04923100, June 10, 2021).

Small cell carcinoma as an independent prognostic factor for cervical cancer patients: a population-based analysis.

Aim: To compare cervical small cell carcinoma (SmCC) with squamous cell carcinoma (SCC) in patient characteristics and survival outcomes. Methods: Cervical SmCC and SCC patients in Surveillance, Epidemiology, and End Results database from 2004 to 2015 were enrolled. Propensity-score matching analysis (PSM) paired subjects with similar background variables. Cox regression, Kaplan-Meier and stratified analyses were conducted before and after PSM. Results: Cervical SmCC patients showed a higher rate of larger tumor size, advanced grade disease, lymph node involvement and distant metastasis (p < 0.001). Before and after PSM, SmCC histology and advanced Federation International of Gynecology and Obstetrics stages (p < 0.001) were principal prognostic factors of survival, and cervical SmCC was associated with worse survival in all stages (stage I-IV). Conclusion: SmCC was an independent poor prognostic factor in cervical cancer patients.

Functional identification of factor VIII B domain regions in hepatocyte cells.

Factor VIII (FVIII) functions as a cofactor within the intrinsic pathway of blood coagulation in process of FX activation by FIXa, for which deficiency results in the bleeding disorder hemophilia A. The gene of FVIII contains 26 exons that code for a 19 amino acid signal peptide and a 2332 amino acid polypeptide with a domain structure designated A1-A2-B-A3-C1-C2, of which the A domains are homologous with each other, as are the C domains. It has been well-documented that both the domains are the necessary elements for FVIII activities. The B domain is highly glycosylated and has a variable sequence, even among FVIIIs from different species. The B domain plays versatile roles in FVIII lifespan except for coagulation activity, but the functional characteristics of its specific regions remain still obscure. A series of recombinant FVIIIs (rFVIIIs) with B domain truncated were constructed and transiently expressed in hepatocyte cells. Media and cell lysates were collected after 72 h for the analyses of FVIII biosynthesis, secretion, activity and stability in ex vivo plasma relative to the full length wild-type FVIII. Unexpectedly, various regions in B domain exhibited different contribution to these functionalities. The discovery might facilitate the bioengineered rFVIIIs and gene therapeutics.

Long non-coding RNA GAS5 regulates the growth and metastasis of human cervical cancer cells via induction of apoptosis and cell cycle arrest.

Studies have proved the role of GAS5 in the development of different cancers. This study was undertaken to investigate the role and explore therapeutic implications of GAS5 in human cervical cancer. The results showed that GAS5 was significantly (p < 0.05) downregulated in human cervical cancer tissues. The results also showed that cervical cancer progresses with the suppression of GAS5 expression levels. Additionally, the expression of GAS5 was also significantly (p < 0.05) downregulated in human cervical cancer cell lines. Nonetheless, overexpression of GAS5 caused a remarkable decrease in the proliferation of C33A and HeLa cervical cancer cells. The decrease in the proliferation rate was attributed to the induction of apoptosis of C33A and HeLa cells which was accompanied with upregulation of Bax and suppression of Bcl-2. Additionally, GAS5 overexpression also promoted the arrest of C33A and HeLa cells at the G2/M check point of cell cycle via suppression of cyclin B1 and CDK1 expression. The transwell assays showed that GAS5 overexpression significantly (p < 0.05) inhibited the migration and invasion of the C33A and HeLa cervical cancer cells. The bioinformatics analysis as well as the dual luciferase assay showed GAS5 acts as a target of miR-135a. Interestingly, the expression of miR-135a was upregulated in the human cervical cancer cells and its suppression exerted growth inhibitory effects on the C33A and HeLa cells. However, silencing of GAS5 could nullify the effects of miR-135a suppression on the proliferation of C33A and HeLa cells. Taken together, the results of this study point towards the therapeutic implications of GAS5 in the treatment of cervical cancer.

Green Tea Consumption and Esophageal Cancer Risk: A Meta-analysis.

Background: The protective role of green tea against cancer is still unknown.Objectives: To investigate the association between green tea consumption and esophageal cancer risk through meta-analysis.Methods: We searched MEDLINE, EMBASE, Web of Science and Cochrane Library for studies on the relationship between green tea and esophageal cancer risk. We assessed heterogeneity (I2) and publication bias (Begg's and Egger's tests). Pooled relative risks (RRs) or odds ratios (ORs) with 95% confidence intervals (CIs) were calculated using random effects models.Results: A total of 20 studies were included. The RRs for all studies was 0.65 (95% CI: 0.57-0.73), with I2 = 75.3% and P = 0. In the subgroup analysis, the following variables showed marked heterogeneity: Asian (RR: 0.64; 95% CI: 0.56-0.73) and non-Asian countries (RR: 0.74; 95% CI: 0.45-1.03), female (RR: 0.55; 95% CI: 0.39-0.71) and male + female (RR: 0.64; 95% CI: 0.54-0.75), case-control study (RR: 0.62; 95% CI: 0.52-0.71), impact factor >3 (RR: 0.65; 95% CI: 0.56-0.75), impact factor <3 (RR: 0.64; 95% CI: 0.48-0.80), Newcastle-Ottawa Scale >7 (RR: 0.82; 95% CI: 0.66-0.97) and Newcastle-Ottawa Scale ≤7 (RR: 0.59; 95% CI: 0.49-0.68).Conclusion: Green tea consumption could be a protective factor for esophageal cancer.

Risk Factors for Reoperation After Debridement of Acute Pancreatitis.

BACKGROUND: Acute pancreatitis (AP) is a common gastrointestinal disorder with a high mortality rate. This study evaluated the incidence of and risk factors for reoperation after debridement of AP. METHODS: This retrospective study included 168 patients diagnosed with AP who had undergone debridement between January 2007 and December 2017 at our hospital. Patients were divided into single-operation and reoperation groups separately. RESULTS: Sixty-eight (40.24%) patients underwent reoperation after AP debridement. The main procedure for reoperation was debridement of necrosis. In univariate analysis, the risk factors for reoperation included younger age; higher admission temperature and heart rate; higher levels of C-reactive protein (CRP), blood urea nitrogen and creatinine; higher Acute Physiology and Chronic Health Evaluation II score and rate of continuous renal replacement therapy; shorter operation interval; lower postoperative albumin level; and high incidence of preoperative and postoperative complications. Multivariate logistic analysis indicated that independent risk factors for reoperation included higher levels of C-reactive protein and creatinine in admission, preoperative percutaneous catheter drainage, and postoperative complications. CONCLUSIONS: The general characteristics and clinical procedures of patients with AP after debridement might affect prognosis and reoperation. The identification of risk factors could help clinicians to provide specific treatment, better ward management, and stratification of reoperation risk.

Discovery of LY3325656: A GPR142 agonist suitable for clinical testing in human.

The discovery and optimization of a novel series of GPR142 agonists are described. These led to the identification of compound 21 (LY3325656), which demonstrated anti-diabetic benefits in pre-clinical studies and ADME/PK properties suitable for human dosing. Compound 21 is the first GPR142 agonist molecule advancing to phase 1 clinic trials for the treatment of Type 2 diabetes.