A novel presentation of idiopathic adulthood ductopenia (IAD) in a young man.

A 24-years-old male patient was admitted to our institution for intermittent jaundice, fatigue, anorexia and dark urine which occurred six times in the past 8 years (twice in 2019). Liver function test showed elevated levels of bilirubin and liver enzymes. He had no fever, vomiting, abdominal pain or diarrhea, and denied special medication, heredity or family history. He drank alcohol occasionally.

Microbial synthesis of efficient palladium electrocatalyst with high loadings for oxygen reduction reaction in acidic medium.

Whereas limited amount of precious metal adsorbed by bacteria conflicting the needs of high loadings for better catalytic performances, cell disruption technology was adopted to smash Shewanella cells in this work, releasing abundant oxygen functional groups inside the cells for better adsorption of palladium ion. Then palladium catalysts were synthesized in two ways: 1) Pd catalyst supported on carbonized-broken-bacterial (Pd/FHNC) was obtained after direct carbonization and reduction; 2) Electrospinning technology was used to spin the broken Shewanella into fibers, and Pd nanoparticles supported on nitrogen-doped carbon nanofiber (Pd/NCNF) was prepared following carbonization and hydrogen reduction. The as-prepared catalysts exhibit excellent oxygen reduction reaction (ORR) electrocatalytic performance in the acid medium. The mass specific activities at 0.7 V of Pd/FHNC and Pd/NCNF were 0.213 A mg-1 and 0.121 A mg-1 which were 5.92 and 3.36 times than those of commercial Pd/C(0.036 A mg-1) respectively, and they also displayed higher stability than Pd/C. Furthermore, the Pd loadings of Pd/FHNC and Pd/NCNF were 21.52% and 17.13% respectively. An explanation for the improved performance is the co-doping of nitrogen and phosphorus, also the tight integration of Pd and broken-bacterial. Herein, we propose a novel and effective method for synthesis of ORR electrocatalysts.

High expression of FABP4 in colorectal cancer and its clinical significance.

OBJECTIVES: To investigate the relationship between the fatty acid-binding protein 4 (FABP4) and colorectal cancer (CRC). METHODS: Using an enzyme-linked immunosorbent assay (ELISA), we measured the expression of FABP4 in plasma of 50 patients who underwent surgery for CRC from October 2017 to May 2018 and 50 healthy controls. The content of the visceral fat area (VFA) as seen with abdominal computed tomography (CT) scanning was measured by ImageJ software. The expression levels of FABP4, E-cadherin, and Snail proteins in CRC and adjacent tissues were determined by immunohistochemistry. RESULTS: The mean concentration of plasma FABP4 of CRC patients was higher than that of the control group (22.46 vs. 9.82 ng/mL; P<0.05). The concentration of plasma FABP4 was related to the tumor, node, metastatis (TNM) stage and lymph node metastasis and was independent of age, body mass index (BMI), tumor size and location, and the degree of differentiation of CRC. The concentration of plasma FABP4 was positively correlated with high VFA and lipoprotein-a (LPA) (P<0.05); but it was not correlated with total cholesterol (TG), total triglyceride (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), or apolipoprotein AI (Apo-AI). The expression of FABP4 protein in CRC tissues was positively correlated with the degree of CRC differentiation, tumor stage, and lymph node metastasis. The level of FABP4 protein was negatively correlated with E-cadherin protein (r=-0.3292, P=0.0196) and positively correlated with Snail protein (r=0.5856, P<0.0001). CONCLUSIONS: High LPA and VFA were risk factors for increased plasma FABP4 in CRC patients. FABP4 protein was highly expressed in CRC tissues and associated with TNM stage, differentiation, and lymph node metastasis of CRC. The level of FABP4 in CRC tissue was correlated with E-cadherin and Snail expression, suggesting that FABP4 may promote CRC progression related to epithelial-mesenchymal transition (EMT).

FABP4 promotes invasion and metastasis of colon cancer by regulating fatty acid transport.

BACKGROUND: The prognosis of colon cancer is poor for metastasis, while the mechanism, especially adipocytes related, is not yet clear. The purpose of this study is to determine the effects of fatty acid binding protein 4 (FABP4), a transporter for lipids, on colon cancer progression. METHODS: The distribution of lipids and FABP4 was tested in the colon cancer tissues and adjacent normal tissues, and their relationship was also verified in vitro. Experiments about cellular invasion, migration and proliferation were performed to detect the impacts of FABP4 on the biological behaviors of colon cancer, and the positive results were checked in vivo. Meanwhile, the regulatory role of FABP4 in the energy and lipid metabolism was evaluated by the levels of triglyceride, ATP, LDH, glycerol and NEFA. At last, GO and KEGG analysis based on FABP4 overexpressed cells was performed, and the AKT pathway and epithelial-mesenchymal transition (EMT)-related proteins were determined by Western blot. RESULTS: Higher accumulation of lipids and stronger FABP4 transcription were observed in colon cancer tissues. Having been incubated with adipose tissue extract and overexpressed FABP4, colon cancer cells demonstrated enhanced lipid accumulation. In functional experiments, co-culture with adipose tissue extract significantly enhanced the invasion and migration of colon cancer cells, as well as the energy and lipid metabolism, and all these processes were reversed by FABP4 inhibitor. In addition, the metastasis of FABP4-overexpressed colon cancer cells was also significantly enhanced in vitro and in vivo. In terms of mechanism, the bioinformatics analysis showed that FABP4 was enriched in 11 pathways related to metabolic processes in FABP4 overexpressed cells. Finally, FABP4 overexpression improved EMT progression of colon cancer, as evidenced by the upregulation of Snail, MMP-2 and MMP-9, the downregulation of E-cadherin. The expression of p-Akt was also elevated. CONCLUSION: FABP4 overexpression could increase FAs transport to enhance energy and lipid metabolism, and activate AKT pathway and EMT to promote the migration and invasion of colon cancer cells.

Clinical Application of Folate Receptor-Mediated Staining Solution Detection in Cervical Cancer Screening.

OBJECTIVE: Cervical cancer is the fourth most deadly women's cancer worldwide, and regular screening is essential to lower mortality rates. The folate receptor-mediated staining solution detection (FRD) has been suggested to be a rapid and cost-effective screening method. We aim to evaluate the validity of FRD testing in cervical cancer screening. METHODS: A total of 207 participants were enrolled in the study. The validity of screening by FRD, cytology screening, and a HPV test were compared using histopathology as the gold standard. Sensitivity, specificity, positive predictive value, negative predictive value, Kappa value, positive likelihood ratio, negative likelihood ratio, percent agreement, and positive detection rates were compared among the three screening methods. RESULTS: 83(40.1%) participants were diagnosed as NILM, 50(24.15%) were diagnosed as CIN1, and 74(35.74%) were diagnosed as CIN2+. For CIN2+, the detection rates for the FRD, cytology screening, and HPV were 75.68%, 82.09% and 93.22%, respectively. For CIN2+, the sensitivity of HPV testing (93.22%) was significantly higher than that of cytology screening (82.09%) and FRD (75.68%), while the specificity of FRD (63.91%) was higher than that of cytology screening (35.34%) and HPV test (7.56%). The percent agreement and Kappa value of FRD were significantly higher than those of the cytology screening and HPV test. In HPV-HC2+ and ASCUS patients, FRD was associated with a lower false positive rate compared to other screening methods. CONCLUSION: Our study indicates that FRD has a good sensitivity and high specificity in cervical cancer screening, and could be a rapid, valid and cost-effective screening test.

Increased HMGB1 expression correlates with higher expression of c-IAP2 and pERK in colorectal cancer.

The aim of this study was to investigate the relationship between high-mobility group box 1 (HMGB1) and colorectal cancer (CRC).In this prospective study, patients with CRC undergoing primary surgery and healthy subjects (control group) were enrolled from July 2013 to December 2014. The serum HMGB1 concentration and HMGB1 mRNA expression were determined using enzyme-linked immunosorbent assay reverse transcription-polymerase chain reaction, respectively. Immunohistochemical analysis was performed to determine HMGB1, pERK, and c-inhibitor of apoptosis protein 2 (c-IAP2) protein expression levels in the cancer tissues.A total 144 patients with CRC and 50 healthy subjects underwent serum HMGB1 testing. Resected specimens of 50 patients were used for HMGB1 mRNA and protein expression analyses. Mean serum HMGB1 level in the patients with CRC was higher than that of the control group (8.42 µg/L vs 1.79 µg/L, P < .05). Mean serum HMGB1 level in the patients with CRC with distant metastasis was significantly higher than that of the controls (13.32 µg/L vs 7.37 µg/L, P < .05). The HMGB1 mRNA and protein expression levels in the CRC tissues were significantly higher than those in the adjacent normal mucosa. HMGB1 protein expression positively correlated with the lymph node metastasis. There were positive correlations between HMGB1 and c-IAP2 (r = 0.457, P < .05), HMGB1 and pERK (r = 0.461, P < .05), as well as pERK and c-IAP2 (r = 0.399, P < .05).HMGB1 expression in CRC correlates with distant and lymph node metastasis. It may inhibit apoptosis by inducing activation of pERK and c-IAP2.

Effect of operating conditions on hydrothermal liquefaction of Spirulina over Ni/TiO2 catalyst.

In this study, the effects of reaction temperature, holding time, algae/water ratio and catalyst dosage on the yield and quality of bio-oil produced via the HTL of Spirulina were investigated. The maximum bio-oil yield (43.05 wt%) and energy recovery (ER) value (64.62%) were obtained at 260 °C for 30 min, with an algae/water ratio of 1/4 and a catalyst dosage of 5 wt%. The bio-oil samples were characterized by elemental analysis, Gas Chromatography-Mass Spectrometry (GC-MS), Fourier Transform Infrared (FI-IR), and Thermo-gravimetric analysis (TGA). Results indicated that higher heating values (HHVs) of bio-oils were in the range of 27.28-36.01 MJ/kg, and main compounds of bio-oil were amides, esters, nitriles, hydroperoxide and alkanes. Adding of the Ni/TiO2 catalyst can decrease the contents of oxygenated and nitrogenous compounds and promote the formation of desirable components such as esters and alkanes.

Hydrothermal liquefaction of microalgae over transition metal supported TiO2 catalyst.

Hydrothermal liquefaction (HTL) of microalgae Nannochloropsis (NAS) over various transition metal M/TiO2 (M = Fe, Co, Ni, Mo, and Mn) was investigated. Ni/TiO2 was the most effective catalyst to improve the yield and quality of biocrude and the liquefaction conversion. Ni/TiO2 was characterized by XRD, XRF, and XPS. The research of Effect of reaction temperature on HTL of NAS over Ni/TiO2 suggested that 300 °C led to a maximum biocrude yield of 48.23% and the highest liquefaction conversion of 89.28%. Adding Ni/TiO2 catalyst reduced the viscosity and provided more light-fraction in biocrude while brought a slight increase in total acid number (TAN). Gas chromatography-mass spectrometry (GC-MS) analysis demonstrated that adding Ni/TiO2 considerably changed the composition of biocrude and the possible pathways were discussed. Reproduction test showed the Ni/TiO2 has an excellent reproduction ability in HTL of NAS.

Research and progress on ClC­2 (Review).

Chloride channel 2 (ClC-2) is one of the nine mammalian members of the ClC family. The present review discusses the molecular properties of ClC­2, including CLCN2, ClC­2 promoter and the structural properties of ClC­2 protein; physiological properties; functional properties, including the regulation of cell volume. The effects of ClC­2 on the digestive, respiratory, circulatory, nervous and optical systems are also discussed, in addition to the mechanisms involved in the regulation of ClC­2. The review then discusses the diseases associated with ClC­2, including degeneration of the retina, Sjögren's syndrome, age­related cataracts, degeneration of the testes, azoospermia, lung cancer, constipation, repair of impaired intestinal mucosa barrier, leukemia, cystic fibrosis, leukoencephalopathy, epilepsy and diabetes mellitus. It was concluded that future investigations of ClC­2 are likely to be focused on developing specific drugs, activators and inhibitors regulating the expression of ClC­2 to treat diseases associated with ClC­2. The determination of CLCN2 is required to prevent and treat several diseases associated with ClC­2.

Ultra-sensitive and absolute quantitative detection of Cu2+ based on DNAzyme and digital PCR in water and drink samples.

Here, we developed an ultra-sensitive and absolute quantitative detection method of Cu2+ based on DNAzyme and digital PCR. The binding model between DNAzyme and Cu2+ and the influence caused by the additional primer sequence were revealed to ensure quantitation independent of standard curves. The binding model of DNAzyme and Cu2+ showed that one molecular DNAzyme could bind one Cu2+ in the biosensor step. Thus, the final quantitative results, evaluated by three parallels, showed that the limit of quantitation (LOQ) was as low as 0.5pmol, while the sensitivity was evaluated as 50fmol. The specificity evaluation of our methodologies shows that extremely low crossing signal is existed within the non-specific ions. Moreover, the results of practical detection have shown that the quantitative results were stable and accurate among different food substrates. In conclusion, a flexible quantitative detection method with ultra-sensitivity was developed to detect trace amounts Cu2+ within different substrates.

Ultra-sensitive "turn-on" detection method for Hg(2+) based on mispairing biosensor and emulsion PCR.

Sensor-based detection methods have inspired the idea that chemical or physical signals could be converted to nucleic acid signals to be quantitatively detected using a combination of appropriate detection tools. To achieve ultra-sensitive and absolute quantitative detection of mercury ion (Hg(2+)), we have combined a mispairing biosensor for Hg(2+) and emulsion PCR. The parameters that might influence the biosensor step, such as the duration of isothermal amplification and the concentration of the sensor oligonucleotide, have been firstly optimized in our study to achieve the most efficient biosensor detection. The evaluation results of secondary structures between the biosensors with different number of T-Hg-T structures achieved by Circular Dichroism have indicated that the secondary hairpin structure would be varied according to the change of number of T-Hg-T structures, which could influence the quantitative detection results. Further optimization of number of T-Hg-T within the biosensor sequences showed that 5 T-Hg-T structures could generate the most efficient amplification. After the above optimizations, the emulsion PCR has been employed to achieve the absolute quantitation of nucleic acid signals. The final results have shown that the limit of quantitation (LOQ) in our study was as low as 40fmol, and the limit of detection (LOD) was 10fmol. The practical detection tests showed that the quantitative results were stable and accurate for all substrates. In conclusion, by combining a mispairing biosensor with emulsion PCR, we developed a flexible and stable quantitative "turn-on" detection method with ultra-sensitivity that can detect trace amounts Hg(2+) within different substrates.

A highly sensitive and specific method for the screening detection of genetically modified organisms based on digital PCR without pretreatment.

Digital PCR has developed rapidly since it was first reported in the 1990 s. It was recently reported that an improved method facilitated the detection of genetically modified organisms (GMOs). However, to use this improved method, the samples must be pretreated, which could introduce inaccuracy into the results. In our study, we explored a pretreatment-free digital PCR detection method for the screening for GMOs. We chose the CaMV35s promoter and the NOS terminator as the templates in our assay. To determine the specificity of our method, 9 events of GMOs were collected, including MON810, MON863, TC1507, MIR604, MIR162, GA21, T25, NK603 and Bt176. Moreover, the sensitivity, intra-laboratory and inter-laboratory reproducibility of our detection method were assessed. The results showed that the limit of detection of our method was 0.1%, which was lower than the labeling threshold level of the EU. The specificity and stability among the 9 events were consistent, respectively. The intra-laboratory and inter-laboratory reproducibility were both good. Finally, the perfect fitness for the detection of eight double-blind samples indicated the good practicability of our method. In conclusion, the method in our study would allow more sensitive, specific and stable screening detection of the GMO content of international trading products.

Development and application of a quantitative loop-mediated isothermal amplification method for detecting genetically modified maize MON863.

BACKGROUND: A SYBR Green I-based quantitative loop-mediated isothermal amplification (LAMP) assay was developed for the rapid detection of genetically modified maize MON863. A set of primers was designed based on the integration region of the Cry3Bb1 and tahsp17 genes. RESULTS: The qualitative and quantitative reaction conditions (dNTPs, betaine, primers, Mg(2+), Bst polymerase, temperature, reaction time) were optimized. The concentrations of Mg(2+) and betaine were found to be important to the LAMP assay. The detection limits of both qualitative and quantitative LAMP for MON863 were as low as 4 haploid genomic DNA, and the LAMP reactions can be completed within 1 h at an isothermal temperature of 65 °C. CONCLUSION: The results of this study demonstrate that this new SYBR Green I-based quantitative LAMP assay system is reliable, sensitive and accurate.

Restriction enzyme cutting site distribution regularity for DNA looping technology.

The restriction enzyme cutting site distribution regularity and looping conditions were studied systematically. We obtained the restriction enzyme cutting site distributions of 13 commonly used restriction enzymes in 5 model organism genomes through two novel self-compiled software programs. All of the average distances between two adjacent restriction sites fell sharply with increasing statistic intervals, and most fragments were 0-499 bp. A shorter DNA fragment resulted in a lower looping rate, which was also directly proportional to the DNA concentration. When the length was more than 500 bp, the concentration did not affect the looping rate. Therefore, the best known fragment length was longer than 500 bp, and did not contain the restriction enzyme cutting sites which would be used for digestion. In order to make the looping efficiencies reach nearly 100%, 4-5 single cohesive end systems were recommended to digest the genome separately.

Single universal primer multiplex ligation-dependent probe amplification with sequencing gel electrophoresis analysis.

In this study, a novel single universal primer multiplex ligation-dependent probe amplification (SUP-MLPA) technique that uses only one universal primer to perform multiplex polymerase chain reaction (PCR) was developed. Two reversely complementary common sequences were designed on the 5' or 3' end of the ligation probes (LPs), which allowed the ligation products to be amplified through only a single universal primer (SUP). SUP-MLPA products were analyzed on sequencing gel electrophoresis with extraordinary resolution. This method avoided the high expenses associated with capillary electrophoresis, which was the commonly used detection instrument. In comparison with conventional multiplex PCR, which suffers from low sensitivity, nonspecificity, and amplification disparity, SUP-MLPA had higher specificity and sensitivity and a low detection limit of 0.1 ng for detecting single crop species when screening the presence of genetically modified crops. We also studied the effect of different lengths of stuffer sequences on the probes for the first time. Through comparing the results of quantitative PCR, the LPs with different stuffer sequences did not affect the ligation efficiency, which further increased the multiplicity of this assay. The improved SUP-MLPA and sequencing gel electrophoresis method will be useful for food and animal feed identification, bacterial detection, and verification of genetic modification status of crops.