Gallbladder carcinosarcoma with a poor prognosis: A case report.

BACKGROUND: Carcinosarcoma of the gallbladder is a rare malignant tumor with a very poor prognosis. To date, only approximately 100 patients have been reported in the English literature. The prognosis of this tumor type is poor, the preoperative diagnosis is difficult, and there is a possibility of a misdiagnosis. We present an unsuccessful case of carcinosarcoma of the gallbladder with a preoperative misdiagnosis and rapid early postoperative recurrence. Therefore, we have a deeper understanding of the poor prognosis of gallbladder carcinosarcoma (GBC) patients. CASE SUMMARY: The patient is a 65-year-old male. He was admitted to the hospital because of right upper abdomen distending pain and discomfort for half a month. Abdominal magnetic resonance imaging revealed a polycystic mass in the right lobe of the liver and the fossa of the gallbladder. After admission, the patient was diagnosed with a liver abscess, which was treated by abscess puncture drainage. Obviously, this treatment was unsuccessful. Hepatectomy and cholecystectomy were performed one month after the puncture. Postoperative pathologic examination revealed carcinosarcoma of the gallbladder, and the resected specimen contained two tumor components. One month after surgery, the patient's tumor recurred in situ and started to compress the duodenum, resulting in duodenal obstruction and bleeding. The treatment was not effective. The patient died of gastrointestinal hemorrhage and hypovolemic shock. CONCLUSION: Carcinosarcoma of the gallbladder is a rare malignant tumor that is easily misdiagnosed preoperatively and has a poor prognosis.

Enhanced reproductive toxicity of photodegraded polylactic acid microplastics in zebrafish.

Microplastics are widely used due to their numerous advantages. However, they can have detrimental effects on marine ecosystems. When microplastics enter the ocean, they can be absorbed by marine organisms, leading to toxic effects. Additionally, the transformation of microplastics during natural degradation can alter their toxicity, necessitating further investigation. Polylactic acid (PLA) biodegradable plastics are commonly used, yet research on their toxicity, particularly their reproductive effects on aquatic organisms, remains limited. In this study, we conducted photodegradation of PLA using potassium persulfate as a catalyst to simulate natural degradation conditions. Our objective was to assess the reproductive toxicity of photodegraded PLA microplastics on zebrafish. The results revealed that photodegraded PLA exhibited elevated reproductive toxicity, resulting in abnormal oocyte differentiation, disruption of sexual hormone levels, and alterations in ovarian tissue metabolism. Metabolomics analysis indicated that both unphotodegraded PLA (UPLA) and photodegraded PLA (DPLA) disrupted oxidative stress homeostasis in zebrafish ovarian tissue by influencing pathways such as purine metabolism, phenylalanine metabolism, glutathione metabolism, and riboflavin metabolism. Furthermore, the DPLA treatment induced abnormal biosynthesis of taurocholic acid, which was not observed in the UPLA treatment group. Importantly, the DPLA treatment group exhibited more pronounced effects on offspring development compared to the UPLA treatment group, characterized by higher mortality rates, inhibition of embryo hatching, accelerated heart rates, and reduced larval body length. These findings underscore the varying levels of toxicity to zebrafish ovaries before and after PLA photodegradation, along with evidence of intergenerational toxicity.

Novel role of immune-related non-coding RNAs as potential biomarkers regulating tumour immunoresponse via MICA/NKG2D pathway.

Major histocompatibility complex class I related chain A (MICA) is an important and stress-induced ligand of the natural killer group 2 member D receptor (NKG2D) that is expressed in various tumour cells. Given that the MICA/NKG2D signalling system is critically embedded in the innate and adaptive immune responses, it is particularly involved in the surveillance of cancer and viral infections. Emerging evidence has revealed the important roles of non-coding RNAs (ncRNAs) including microRNAs (miRNAs), long noncoding RNAs (lncRNAs) and circular RNAs (circRNAs) in different cancer types. We searched for all relevant publications in the PubMed, Scopus and Web of Science database using the keywords ncRNA, MICA, NKG2D, cancer, and miRNAs. All relevant studies published from 2008 to the 2023 were retrieved and collated. Notably, we found that miRNAs can target to NKG2D mRNA and MICA mRNA 3'-untranslated regions (3'-UTR), leading to translation inhibition of NKG2D and MICA degradation. Several immune-related MICA/NKG2D pathways may be dysregulated in cancer with aberrant miRNA expressions. At the same time, the competitive endogenous RNA (ceRNA) hypothesis holds that circRNAs, lncRNAs, and mRNAs induce an abnormal MICA expression by directly targeting downstream miRNAs to mediate mRNA suppression in cancer. This review summarizes the novel mechanism of immune escape in the ncRNA-related MICA/NKG2D pathway mediated by NK cells and cancer cells. Moreover, we identified the miRNA-NKG2D, miRNA-MICA and circRNA/lncRNA/mRNA-miRNA-mRNA/MICA axis. Thus, we were particularly concerned with the regulation of mediated immune escape in the MICA/NKG2D pathway by ncRNAs as potential therapeutic targets and diagnostic biomarkers of immunity and cancer.

Mechanism of PAX6 overexpression in inhibiting the growth of hepatocellular carcinoma cells and promoting the killing ability of the natural killer cells. / 过表达PAX6抑制肝癌细胞生长和促进自然杀伤细胞杀伤能力的机制.

OBJECTIVES: Paired box gene 6 (PAX6) plays a major role in the regulation of embryonic development. Abnormal expression of PAX6 is associated with the development of various tumors. PAX6 can play a role in promoting or suppressing cancer in different tumors. This study aim to observe the effect of overexpression of PAX6 on the growth of hepatocellular carcinoma cells, and the killing of hepatocellular carcinoma cells via natural killer (NK) cell and the possible mechanism. METHODS: The protein levels of PAX6, soluble major histocompatibility complex class I-like protein A (sMICA) and soluble UL16 binding protein 2 (sULBP2) in peripheral blood from 68 cases of hepatocellular carcinoma (HCC) patients and 10 healthy volunteers were detected by ELISA. Hepatocellular carcinoma cell line (HepG2, LM3) and human normal liver cells (LO2) were cultured at 37 ℃ and 5% CO2 condition in vitro. The PAX6 overexpressed plasmid (PAX6-OE) and empty vector (NC) were transferred into HepG2 and LM3 cells to construct stable cell lines. The mRNA and protein expression levels of PAX6 in HepG2 and LM3 cells were detected by real-time PCR, Western blotting and immunofluorescence, respectively. PAX6 was overexpressed in HepG2 and LM3 cells, the cell growth and migration ability were detected by CCK-8 method and cell scratch assay, and the levels of sMICA and sULBP2 in the supernatant were detected by ELISA. Matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9) and disintegrin and metalloproteinase 10 (ADAM10) in HepG2 and LM3 cells were detected by Western blotting. The killing ability of NK cells against these 2 HCC cells was detected by flow cytometry. RESULTS: Compared with the healthy volunteers, the expressions of PAX6 in the HCC patients were significantly decreased (P=0.002), while the expression of sMICA and sULBP2 were significantly increased (P=0.004 and P<0.001, respectively). Real-time PCR and Western blotting results showed that compared with LO2 cells, mRNA and protein expressions of PAX6 in HepG2 and LM3 cells were significantly decreased (all P<0.05). Immunofluorescence results also showed that the expressions of PAX6 in HepG2 and LM3 were lower than those of LO2 cells. Compared with the NC group, the ability of proliferation and migration of HepG2 and LM3 cells were decreased (both P<0.05). The protein expressions of MMP2, MMP9 and ADAM10 in HepG2 and LM3 cells in the PAX6-OE group were significantly decreased, and the levels of sMICA and sULBP2 in superneant of HepG2 and LM3 cells in the PAX6-OE group were significantly lower than those in the NC group (all P<0.05). Flow cytometry results showed that compared with the NC group, the proportion of NK cells killing HepG2 and LM3 cells in PAX6-OE group was significantly increased (both P<0.05). CONCLUSIONS: The expression of PAX6 is decreased in serum of HCC patients and hepatocellular carcinoma cell lines. Overexpression of PAX6 can inhibit the growth of hepatocellular carcinoma cells, enhance the killing efficiency of NK cells against hepatoma cells. The mechanism is related to the inhibition of the expression of metalloproteinase via PAX6 and the decrease of the secretion levels of sMICA and sULBP2.

The combined toxic effects of polystyrene microplastics and different forms of arsenic on the zebrafish embryos (Danio rerio).

Microplastics have been widely studied for their ability to adsorb heavy metals. In the natural environment, arsenic exists in different forms and its toxicity depends mainly on its form and concentration. However, different forms of arsenic combined with microplastics have yet to be explored for their biological hazards. This study was conducted to reveal the adsorption mechanism of different forms of arsenic onto PSMP and to study the effects of PSMP on the tissue accumulation and developmental toxicity of different forms of arsenic in zebrafish larvae. As a result, the absorbing ability of PSMP for As(III) was 35 times higher than that of DMAs, in which hydrogen bonding plays an important role in the adsorption process. In addition, the adsorption kinetics of As(III) and DMAs on PSMP were in good agreement with the pseudo-second-order kinetic model. Furthermore, PSMP reduced the accumulation of As(III) early in zebrafish larvae development, thereby increasing hatching rates compared with the As(III)-treated group, whereas PSMP had no significant effect on DMAs accumulation in zebrafish larvae, but decreased hatching rates compared with the DMAs-treated group. In addition, except for the microplastic exposure group, the other treatment groups could lead to a decrease in the heart rate of zebrafish larvae. Both PSMP+As(III) and PSMP+DMAs exhibited aggravated oxidative stress compared with PSMP-treated group, but PSMP+As(III) caused more severe oxidative stress at later stages of zebrafish larvae development. Moreover, specific metabolic differences (e.g., AMP, IMP, and guanosine) were produced in the PSMP+As(III) exposure group, which would mainly affect purine metabolism and promoted specific metabolic disturbances. However, PSMP+DMAs exposure shared metabolic pathways altered by PSMP and DMAs, indicating an independent effect of these two chemicals. Taken together, our findings emphasized that the combined toxicity of PSMP and different forms of arsenic posed a health risk that cannot be ignored.

Analysis of Antibodies Induced after SARS-CoV-2 Vaccination Using Antigen Coded Bead Array Luminex Technology.

Objectives. Since the outbreak of SARS-CoV-2 in late 2019, nearly 12.2 billion doses of the COVID-19 vaccine have been administered worldwide; however, the humoral immune responses induced by different types of vaccines are yet to be fully validated. Methods. We analyzed antibody levels in 100 serum samples after vaccination with different types of COVID-19 vaccines and their reactivity against the RBD antigen of Delta and Omicron variants using a bead-based microarray. Results. Elevated levels of anti-wild-type (WT)-RBD IgG and anti-WT-NP IgG were detected in participants who received two doses of the inactivated vaccines (CoronaVac or BBIBP-CorV) and three doses of the recombinant spike protein vaccine (ZF2001), indicating that antibody responses to SARS-CoV-2 were generated regardless of the vaccine administered. We found highly correlated levels of serum anti-RBD IgG and anti-NP IgG (r = 0.432, p < 0.001). We observed that the antibodies produced in vivo after COVID-19 vaccination still reacted with variants of SARS-CoV-2 (p < 0.0001). Conclusions. Our results show that high levels of specific antibodies can be produced after completion of COVID-19 vaccination (two doses of the inactivated vaccines or three doses of ZF2001), with some degree of cross-reactivity to the RBD antigen of Delta and Omicron variants, and provide an accessible and practical experimental method for post-vaccination antibody detection.

Extracorporeal Membrane Oxygenation in Acute Respiratory Distress Syndrome Caused by Anhydrous Ammonia Burns: Two Case Reports.

Acute anhydrous ammonia burns are relatively rare but lethal and often occur as a mass occupational incident worldwide. Anhydrous ammonia mainly leads to severe inhalation injury and skin/mucosa wound because of its high water solubility and strong alkalinity. Acute respiratory distress syndrome (ARDS) induced by inhalation injury is the main cause of death. Extracorporeal membrane oxygenation (ECMO), also known as extracorporeal life support, has been recommended as the salvage treatment for severe ARDS based on low-level evidence. However, the application of ECMO in ammonia burns is still limited. Here, we presented two cases of anhydrous ammonia burn patients, one 62-year-old man with 15% total body surface area (TBSA) and one 47-year-old man with 27% TBSA, accompanying severe inhalation injury. They both developed severe ARDS and started vv ECMO on 3, 6, and 15 days after injury, respectively. ECMO lasted 118, 247, and 72 h, respectively. All ECMO were successfully weaned off although only one patient survived. Meanwhile, one patient had the coagulopathy complication of ECMO, mainly bleeding, deep vein thrombosis, and hemolysis. In conclusion, this report provided evidence for use of ECMO as supportive care in ammonia burn patients with severe ARDS.

Recent Progress in Biosensors for Detection of Tumor Biomarkers.

Cancer is a leading cause of death worldwide, with an increasing mortality rate over the past years. The early detection of cancer contributes to early diagnosis and subsequent treatment. How to detect early cancer has become one of the hot research directions of cancer. Tumor biomarkers, biochemical parameters for reflecting cancer occurrence and progression have caused much attention in cancer early detection. Due to high sensitivity, convenience and low cost, biosensors have been largely developed to detect tumor biomarkers. This review describes the application of various biosensors in detecting tumor markers. Firstly, several typical tumor makers, such as neuron-specific enolase (NSE), carcinoembryonic antigen (CEA), prostate-specific antigen (PSA), squamous cell carcinoma antigen (SCCA), carbohydrate, antigen19-9 (CA19-9) and tumor suppressor p53 (TP53), which may be helpful for early cancer detection in the clinic, are briefly described. Then, various biosensors, mainly focusing on electrochemical biosensors, optical biosensors, photoelectrochemical biosensors, piezoelectric biosensors and aptamer sensors, are discussed. Specifically, the operation principles of biosensors, nanomaterials used in biosensors and the application of biosensors in tumor marker detection have been comprehensively reviewed and provided. Lastly, the challenges and prospects for developing effective biosensors for early cancer diagnosis are discussed.

A Soluble NK-CAR Mediates the Specific Cytotoxicity of NK Cells toward the Target CD20+ Lymphoma Cells.

The structures of chimeric antigen receptors (CARs) currently designed for natural killer (NK) cells are mostly based on knowledge gained about CAR-T cells. Although these CAR-NK cells have shown promising effects, there are still many limitations to their application. In this study, we designed a soluble NK-CAR since the membrane protein NKG2D expressed by NK cells can directly trigger NK cell cytotoxicity by binding with the ligand MICA. This CAR is composed of three segments: the extracellular domain of MICA, an anti-CD20 single-chain variable fragment (anti-CD20 ScFv), and a human IgG Fc component. The nucleotide sequence of the soluble NK-CAR was cloned into a eukaryotic expression vector and expressed in suspension HEK293 cells, and the recombinant NK-CAR protein was then purified in a Staphylococcus aureus protein A column. The novel NK-CAR exhibited bifunctional activity, recognizing both the CD20 antigen of target cells and the NKG2D receptor of NKL cells. The NK-CAR activated the NKG2D receptor signaling pathway, causing NKL cells to express CD107a and secrete interferon-gamma. The soluble NK-CAR mediated the NKL cell killing of CD20+ Daudi cells in vitro, with a 1 µg/mL concentration inducing the maximum killing effect. Moreover, 51.7% (p < 0.01) of Daudi cells were killed at the effector-to-target ratio of 10:1. In the presence of recombinant rMICA and NKG2D-Ig proteins, this killing effect was reduced to 30% (P < 0.01) owing to competitive interference. Our results highlight the clinical application potential of this novel immunotherapy for killing target tumor cells.

Posttransplant-Alloantibodies Against MICA Antigens Associated With Decreased Long-Term Allograft Survival of Kidney Transplant Recipients.

BACKGROUND: Previous evidence showed that antibodies against major histocompatibility complex class I-related chain A (MICA) could lead to antibody-mediated rejection in kidney transplantation in case where the patients had no alloantibodies against HLA. However, the effects of posttransplant anti-MICA antibodies on long-term renal allograft survival and function remained unsettled. We tested the posttransplant anti-MICA antibodies in 150 kidney transplant patients. The aim of this study was to compare the long-term graft survival and function between patients who were MICA positive and those who were negative. METHODS: The posttransplant serum samples from 150 patients receiving kidney transplantation in our center from 2012 to 2013 were tested for MICA antibodies and HLA antibodies by Luminex single antigen array technology. Graft survival and function were followed up for a mean time of 74.2 months. The research was conducted in accordance with the Helsinki Congress and the Declaration of Istanbul. RESULTS: Of the 150 patients, 38 (25.3%) were sensitized against MICA after transplantation. The anti-MICA antibodies-positive (anti-MICA+) group had a worse long-term renal allograft survival than that of anti-MICA-negative (anti-MICA-) group (P = .029), even when stratified by posttransplant HLA sensitization status or donor source. Anti-MICA antibodies also had a detrimental impact on renal allograft function, but only at 1 year posttransplantation (estimated glomerular filtration rates at 1 year: anti-MICA+ 66.6 mL/min/1.73 m2 vs anti-MICA- 78.7 mL/min/1.73 m2; P = .023). CONCLUSION: Posttransplant anti-MICA antibodies were associated with decreased long-term renal allograft survival and short-term renal allograft function.

Detection of Exogenous γ-Hydroxybutyric Acid in Rat Blood Exosomes.

OBJECTIVES: To find a method to distinguish exogenous gamma-hydroxybutyrate (GHB) from endogenous GHB by establishing ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) based on exosome for quantitative detection of GHB in the rat blood. METHODS: Adult male SD rats were divided into 1 h, 5 h, 10 h administration group and control group. After 1 h, 5 h and 10 h of single precursor of GHB gamma-butyrolactone (GBL) intraperitoneal injection in administration groups, 5 mL blood was collected from the abdominal aorta. Meanwhile, the control group was given a same dose of normal saline, and 5 mL blood was collected at 1 h. Among the 5 mL blood, 0.5 mL was directly detected by HPLC-MS after pretreatment, and exosomes were extracted from the remaining blood by differential centrifugation and detected. RESULTS: The concentration of GHB in the control group was (87.36±33.48) ng/mL, and the concentration with administration at 1 h, 5 h and 10 h was (110 400.00±1 766.35) ng/mL, (1 479.00±687.01) ng/mL and (133.60±12.17) ng/mL, respectively. The results of exosome detection showed that no peak GHB signal was detected in the control group and the 10 h administration group, and the concentrations of GHB at 1 h and 5 h administration groups were (91.47±33.44) ng/mL and (49.43±7.05) ng/mL, respectively. CONCLUSIONS: GHB was detected in blood exosome by UPLC-MS, which indicated that exogenous GHB could be detected in plasma exosomes, while endogenous GHB could not be detected, suggesting that this method may be used as a basis to determine whether there is exogenous drug intake.

Qualitative and quantitative analysis of the major components in Qinghao Biejia decoction by UPLC-Orbitrap Fusion-MS/MS and UPLC-QQQ-MS/MS and evaluation of their antibacterial activities.

OBJECTIVE: In the present study, the chemical components of Qinghao Biejia decoction (QBD) were qualitatively and quantitatively analyzed using UPLC-Orbitrap Fusion-MS/MS and UPLC-QQQ-MS/MS techniques, followed by identification of each component's origin and evaluation of the antibacterial activity of QBD and its components. METHODS: High-resolution mass spectrometry was used to obtain information on the precise molecular weight, retention time, and fragmentation ion peaks of the compounds used to identify the components of QBD and establish a method for their quantification. In vitro assays including determination of the minimal inhibitory concentration and growth curves were used to assess the antibacterial activity of QBD and its components. RESULTS: A total of 39 components, including fatty acids, phenolic acids, amino acids, flavonoids, coumarins, terpenoids, and alkaloids, were identified by UPLC-Orbitrap Fusion-MS/MS. A high-performance analytical method was also established to quantify 12 components of QBD. The content of mangiferin was relatively high (estimated to be 814 µg/g). The results of the antibacterial assays indicated that mangiferin exhibits antibacterial effects against two strains causing respiratory tract infections. CONCLUSIONS: The present study suggests that mangiferin may serve as a natural compound which shows high antibacterial activity. The results can aid the discovery and analysis of the active antimicrobial components present in QBD and further provide a reference for quality assessment of multi-component herbal prescriptions.

Proteome-wide analysis of lysine ß-hydroxybutyrylation in the myocardium of diabetic rat model with cardiomyopathy.

Lysine ß-hydroxybutyrylation (kbhb), a novel modification of lysine residues with the ß-hydroxybuty group, is associated with ketone metabolism in numerous species. However, its potential role in diabetes, especially in diabetic cardiomyopathy (DCM), remains largely unexplored. In this study, using affinity enrichment and liquid chromatography-mass spectrometry (LC-MS/MS) method, we quantitatively analyze the kbhb residues on heart tissues of a DCM model rat. A total of 3,520 kbhb sites in 1,089 proteins were identified in this study. Further analysis showed that 336 kbhb sites in 143 proteins were differentially expressed between the heart tissues of DCM and wild-type rats. Among them, 284 kbhb sites in 96 proteins were upregulated, while 52 kbhb sites in 47 proteins were downregulated. Bioinformatic analysis of the proteomic results revealed that these kbhb-modified proteins were widely distributed in various components and involved in a wide range of cellular functions and biological processes (BPs). Functional analysis showed that the kbhb-modified proteins were involved in the tricarboxylic acid cycle, oxidative phosphorylation, and propanoate metabolism. Our findings demonstrated how kbhb is related to many metabolic pathways and is mainly involved in energy metabolism. These results provide the first global investigation of the kbhb profile in DCM progression and can be an essential resource to explore DCM's pathogenesis further.

A Review of Detection of Antibiotic Residues in Food by Surface-Enhanced Raman Spectroscopy.

Antibiotics, as veterinary drugs, have made extremely important contributions to disease prevention and treatment in the animal breeding industry. However, the accumulation of antibiotics in animal food due to their overuse during animal feeding is a frequent occurrence, which in turn would cause serious harm to public health when they are consumed by humans. Antibiotic residues in food have become one of the central issues in global food safety. As a safety measure, rapid and effective analytical approaches for detecting these residues must be implemented to prevent contaminated products from reaching the consumers. Traditional analytical methods, such as liquid chromatography, liquid chromatography mass spectrometry, and capillary electrophoresis, involve time-consuming sample preparation and complicated operation and require expensive instrumentation. By comparison, surface-enhanced Raman spectroscopy (SERS) has excellent sensitivity and remarkably enhanced target recognition. Thus, SERS has become a promising alternative analytical method for detecting antibiotic residues, as it can provide an ultrasensitive fingerprint spectrum for the rapid and noninvasive detection of trace analytes. In this study, we comprehensively review the recent progress and advances that have been achieved in the use of SERS in antibiotic residue detection. We introduce and discuss the basic principles of SERS. We then present the prospects and challenges in the use of SERS in the detection of antibiotics in food. Finally, we summarize and discuss the current problems and future trends in the detection of antibiotics in food.

Expression and purification of Rv2654 recombinant protein from Mycobacterium tuberculosis and its characteristics of immune response. / ç»“æ ¸åˆ†æžæ†èŒæŠ—åŽŸRv2654é‡ç»„è›‹ç™½çš„è¡¨è¾¾å’Œçº¯åŒ–åŠå ¶è¯±å¯¼çš„å ç–«åº”ç­”ç‰¹å¾.

OBJECTIVES: With the decreased protective effect of Bacille Calmette-Guérin (BCG) vaccine, widespread drug-resistant strains of tuberculosis as well as the lack of effective vaccines, global morbidity and mortality of tuberculosis remains high. The purpose of this study is to evaluate the potential of Mycobacterium tuberculosis antigen Rv2654 as a candidate vaccine against tuberculosis, and to verify the characteristics of cellular and humoral immune responses in mice induced by this protein. METHODS: Isopropyl-beta-D-thiogalactoside (IPTG) was added to induce the expression of Rv2654 recombinant protein in the logarithmic growth stage of bacteria. The recombinant protein was purified by affinity chromatography and identified by Western blotting. The immunoreactivity of Rv2654 recombinant protein with human sera was detected by ELISA. After immunization with Rv2654 recombinant protein, the levels of Th1/Th2 cytokines in peripheral blood of mice was measured using cytokine magnetic bead arrays, and the T and B lymphocyte subsets in spleen of mice was analyzed by flow cytometry. RESULTS: The recombinant protein Rv2654 was successfully induced and expressed. The ELISA data showed that the recombinant protein Rv2654 was responsive to the serum of BCG-inoculated patients or active pulmonary tuberculosis patients. In immunized mice with recombinant protein Rv2654, the expression of IFN-γ, TNF-α, IL-2, IL-4, IL-6, IL-10 and other cytokines in peripheral blood was elevated (all P<0.05). Flow cytometry analysis showed that the recombinant protein significantly stimulated the differentiation of CD4+T cells and CD8+T cells into effector T cells, and this effect was more obvious when combined with BCG. The recombinant protein Rv2654 also stimulated the activation and proliferation of B cells and their differentiation into memory cells. However, less plasma cells were produced. CONCLUSIONS: Rv2654 protein could induce the cell immune responses and it has good binding ability with serum of BCG-inoculated patients and active pulmonary tuberculosis patients, suggesting that it may serve as a good candidate antigen for tuberculosis immunological prophylaxis and diagnostic vaccines.

Arginine methylation of METTL14 promotes RNA N6-methyladenosine modification and endoderm differentiation of mouse embryonic stem cells.

RNA N6-methyladenosine (m6A), the most abundant internal modification of mRNAs, plays key roles in human development and health. Post-translational methylation of proteins is often critical for the dynamic regulation of enzymatic activity. However, the role of methylation of the core methyltransferase METTL3/METTL14 in m6A regulation remains elusive. We find by mass spectrometry that METTL14 arginine 255 (R255) is methylated (R255me). Global mRNA m6A levels are greatly decreased in METTL14 R255K mutant mouse embryonic stem cells (mESCs). We further find that R255me greatly enhances the interaction of METTL3/METTL14 with WTAP and promotes the binding of the complex to substrate RNA. We show that protein arginine N-methyltransferases 1 (PRMT1) interacts with and methylates METTL14 at R255, and consistent with this, loss of PRMT1 reduces mRNA m6A modification globally. Lastly, we find that loss of R255me preferentially affects endoderm differentiation in mESCs. Collectively, our findings show that arginine methylation of METTL14 stabilizes the binding of the m6A methyltransferase complex to its substrate RNA, thereby promoting global m6A modification and mESC endoderm differentiation. This work highlights the crosstalk between protein methylation and RNA methylation in gene expression.

Fatal poisoning by accidental ingestion of the "heartbreak grass" (Gelsemium elegans) verified by toxicological and medico-legal analyses.

We present a case of fatal poisoning from accidental ingestion of Gelsemium elegans (G. elegans), a rarely toxic plant. A 41-year-old man was found dead, at his home, 6 h after drinking homemade herbal liqueur during lunch. Autopsy and routine toxicological analyses identified neither significant pathological findings nor routine poisons. However, a local botanist revealed that the homemade herbal liqueur contained G. elegans, a poisonous plant specific to Asia. To ascertain whether the decedent had ingested G. elegans, we performed liquid chromatography-mass spectrometry (LC-MS) and found two alkaloids (gelsemine and koumine) in his blood, gastric contents, as well as the suspected herbal liqueur. The cause of death was therefore confirmed to be G. elegans poisoning. Case reports of fatal poisoning due to ingestion of G. elegans are quite rare in English. Therefore, the present case broadens the scope on the possibility of death due to ingestion of G. elegans for forensic pathologists and toxicologists.

[Overexpression of lncRNA MEG3 inhibits proliferation and invasion of glioblastoma U251 cells in vitro by suppressing HIF1α expression].

OBJECTIVE: To investigate the effects of overexpression of long noncoding RNA (lncRNA) MEG3 on the proliferation and invasion of glioblastoma U251 cells by suppressing the expression of hypoxia inducible factor 1α(HIF1α). METHODS: The expression of lncRNA MEG3 and HIF1α mRNA were examined in human fetal glial cells (HFGCs) and U251 cells using realtime quantitative PCR (qRT-PCR), and the expression of HIF1α protein was detected with Western blotting.U251 cells in normal culture or transfected with pcDNA3.1 vector (NC group) or pcDNA3.1-MEG3 vector via lipofectamine2000 were exposed to hypoxia for 12h, and the expressions of HIF1α mRNA and protein were detected with qRT-PCR and Western blotting, respectively.MTT assay and Transwell assay were employed to examine the influence of MEG3 overexpression on the proliferation and invasion of U251 cells. RESULTS: The expression of MEG3 was significantly lower and HIF1α mRNA and protein expressions were significantly higher in U251 cells than in HFGCs (P < 0.05).In U251 cells, overexpression of MEG3 significantly decreased the mRNA and protein expressions of HIF1α(P < 0.05).Hypoxic exposure for 12h also resulted in significantly lowered expression of HIF1α protein in U251 cells (P < 0.05).Overexpression of MEG3 obviously suppressed the proliferation and invasiveness of U251 cells (P < 0.05). CONCLUSIONS: MEG3 overexpression inhibits the proliferation and invasion of U251 cells through suppressing the expression of HIF1α mRNA and protein, suggesting that MEG3 may serve as a potential therapeutic target for glioblastomas.

MOF-derived zirconia-supported Keggin heteropoly acid nanoporous hybrids as a reusable catalyst for methyl oleate production.

In this study, a series of nanoporous HSiW@ZrO2 hybrids were synthesized using a zirconium metal-organic framework UiO-66 as a precursor towards biodiesel production. The structural and morphological properties of the obtained hybrids were characterized by the wide-angle XRD, FTIR, SEM, TEM, N2 adsorption/desorption, and NH3-TPD methods. Moreover, their catalytic activity in terms of calcination temperature during preparation was investigated, and the HSiW@ZrO2 hybrids calcinated at 300 °C exhibited the highest activity and the oleic acid (OA) conversion of 94.0% owing to the presence of the relatively high surface area, appropriate pore size and strong acidity. It was also revealed that the hybrids maintained as high as 82.0% even after nine cycles. Intriguingly, the nanoporous catalysts were found to exhibit excellent catalytic activity towards the esterification of the high acid value of Jatropha curcas oil.

Construction of a Keggin heteropolyacid/Ni-MOF catalyst for esterification of fatty acids.

This work reports the one-pot solvothermal synthesis of a Keggin heteropolyacid (phosphomolybdic acid, tungstophosphoric acid, or silicotungstic acid) immobilized on Ni-MOF composite catalysts for esterification of fatty acids, and the composites were further analyzed by XRD, FTIR, NH3-TPD, SEM, TEM, N2 adsorption/desorption, and XPS. Among the contrastive syntheses (i.e., HPW/Ni-MOF, HSiW/Ni-MOF, and HPMo/Ni-MOF), HPMo/Ni-MOF exhibits the most active catalyst toward fatty acids esterification, and the characterization results also revealed that HPMo/Ni-MOF has a strong acidity, large specific surface area, and appropriate average pore size. More significantly, this catalyst exhibits a good catalytic performance (86.1% conversion) during esterification under the optimized reaction conditions, and the HPMo/Ni-MOF catalyst can remain stable after the tenth cycle with a conversion of 73.5%. Intriguingly, the esterification reaction kinetics was studied, and the activation energy was found to be 64.6 kJ mol-1. The results indicated that the esterification of fatty acids using the HPMo/Ni-MOF catalyst is a chemically controlled reaction.