In order to investigate how macrophages promote lipid droplet hoarding in hepatocellular carcinoma cells and to delve into the roles of key metabolic enzymes of lipid droplets in the malignant biology of hepatocellular carcinoma cells, the present study was conducted to induce the generation of Hepa1-6 lipid droplets (LDs) using supernatants from tumor-associated macrophages (TAMs), and found that interleukin-10 (IL-10) in TAMs was found to promote the accumulation of lipid droplets. Acetyl-CoA carboxylase alpha (ACC1) is one of the key enzymes for fatty acid synthesis and influences the development of hepatocellular carcinoma. In this study, ACC1 was found to be highly expressed in LDhigh Hepa1-6, and subsequent blockade of ACC1 activity by means of a small molecule inhibitor of ACC1, siRNA interference, and CRISPR-cas9 knockdown was found to reduce the accumulation of Hepa1-6 lipid droplets, as well as to reduce the malignant biological behavior of Hepa1-6 proliferation, and to promote the occurrence of apoptotic events. In summary, IL-10 released by TAMs promoted lipid droplet formation in hepatocellular carcinoma cells, leading to malignant proliferation and apoptosis of hepatocellular carcinoma cells. ACC1 plays a key role in the promotion of lipid droplet accumulation in hepatocellular carcinoma cells by TAMs and may be regulated by IL-10 released by TAMs, and these findings may provide a new target for hepatocellular carcinoma treatment.

Objective To analyze the structures and molecular weight distributions of the capsular polysaccharides from 6 serotypes of pneumococcus .Methods The structures of pneumococcal capsular pol-ysaccharides of 6 serotypes were analyzed by 1 H nuclear magnetic resonance ( NMR) .Chemical shifts of all characteristic protons were investigated to analyze polysaccharide integrity and inter -assay consistency .High performance size exclusion chromatography-multi angle laser light scattering ( HPSEC-MALLS) was used to measure the molecular weights .Results The chemical shifts of all characteristic protons of the pneumococ-cal capsular polysaccharides of 6 serotypes were consistent with the standard chemical shift .The weight-aver-age molecular mass of the pneumococcal capsular polysaccharides ranged from 7.182×104 g/mol(for serotype 19A) to 1.273×106 g/mol(for serotype 9V)examined by HPSEC-MALLS.Conclusion The structures and molecular weight distributions of pneumococcal capsular polysaccharides could be rapidly and effectively ana -lyzed by 1 H NMR and HPSEC-MALLS.Moreover, C-PS and acetate contained in capsular polysaccharides could also be detected .HPSEC-MALLS is an applicable method for the quantitative analysis of molar mass distributions in different serotypes of pneumococcal capsular polysaccharides . Although 1 H NMR and HPSEC-MALLS have been accepted as the quality control measurements by WHO , to use them as the re-placements of the traditional QC method still needs further investigation .

Objective To investigate the molecular size distribution and the structure of group B me-ningococcal capsular polysaccharides for the development of vaccines .Methods The molecular size distribution of group B meningococcal capsular polysaccharides was analyzed by chromatography on a Sepharose CL -4B col-umn.The molecular weight of repeat units were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS).The structural characteristics of group B meningococcal capsular polysaccharides were analyzed by nuclear magnetic resonance ( NMR) based on the chemical shift of all charac-teristic protons by using group C meningococcal capsular polysaccharides and sialic acid as the controls .Results The KD value of group B meningococcal capsular polysaccharides extracted from 15 strains were ranged from 0.60 to 0.76.The molecular weight of repeat units was 284, which was identical to the theoretical value .The group B meningococcal capsular polysaccharides were 2→8 linked homopolymers of sialic acid lacking O-acetyl groups.Conclusion The group B meningococcal capsular polysaccharides had lower molecular weights , which might result in their poor immunogenicity .The structure of group B meningococcal capsular polysaccharides could be quickly and accurately analyzed by NMR technology .

OBJECTIVETo study the relationship between the TCM Syndrome Differentiation-types of congestive heart failure (CHF) and thyroid hormones, including triiodothyronine (T3), thyroxine (T4) and thyroid stimulating hormone (TSH), and atrial natriuretic peptide (ANP), as well as cardiac function parameters, including left ventricular ejection fraction (LVEF), mean velocity of circumferentid fiber shortening (mVcf) and A peak/E peak (A/E).

METHODSOne hundred patients with CHF were divided into 4 Syndrome Differentiation-type groups, their cardiac function parameters, ANP and thyroid hormones were determined and compared with those in the 23 subjects in the control group.

RESULTSIn CHF patients with edema and blood stasis Syndrome type, the level of plasma ANP was significantly higher than that in the control group (P < 0.05); level of T3 was significantly lower than that in the control group and in CHF patients of other three (Xin-qi deficiency, Yin-deficiency and blood stasis) Syndrome groups (P < 0.01, P < 0.01, P < 0.05 and P < 0.01); levels of LVEF and mVcf were significantly lower than those in the other three Syndrome groups (all P < 0.01). Level of T4 in other three Syndrome groups significantly increased than that in the edema and blood stasis Syndrome type. A/E value showed a higher level in patients of all TCM type than that in the control (P < 0.01). Correlation analysis showed that T3 was positively correlated with LVEF and T4 (r = 0.200, P < 0.05, and r = 0.293, P < 0.01), and negatively correlated with ANP (r = -0.263, P < 0.01); T4 was negatively correlated with A/E (r = -0.226, P < 0.05).

CONCLUSIONThe lowering of T3 and T4 and increasing of ANP may be one of the important reasons for lowering of LVEF in CHF patients with edema and blood stasis Syndrome-type. The decrease of T4 may be one of the important reasons for elevation of A/E and aggravation of left ventricular diastolic dysfunction in CHF patients of all the 4 TCM Syndrome-types.

Adult ; Aged ; Atrial Natriuretic Factor ; metabolism ; Diagnosis, Differential ; Female ; Heart Failure ; blood ; physiopathology ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Myocardial Contraction ; Stroke Volume ; physiology ; Thyroid Hormones ; blood ; Thyrotropin ; blood ; Thyroxine ; blood ; Triiodothyronine ; blood ; Ventricular Dysfunction, Left ; physiopathology ; Ventricular Function, Left

Objective: To optimize the existing methods of isolation and purification for exosomes from urine and explore the effects of different storage conditions on the content of exosomal RNA in urine. Methods: The exosomes in human urine samples were extracted by different precipitation method, i.e., precipitation following first concentrating and direct precipitation, respectively, and the separation efficiency and cost of the two methods were compared. ExoQuick-TCTM precipitation kit was used to extract exosomes. Nanoparticle tracking analysis technique (NTA) was used to detect the concentration and particle size distribution of exosome. Dynamic light scattering (DLS) was used to detect the potential of exosome. Transmission electron microscopy (TEM) was used to observe morphology of exosomes. western blot was used to analyze the exosomal marker molecules CD63 and Alix. The extraction method of the precipitation following first concentrating was used to verify the reliability of the optimized method in 10 clinical urine samples . Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of exosomal RNA marker let-7c and PSA mRNA in the urinary exosomes from 20 patients with prostate cancer after repeated freeze-thaw (0 [i.e., fresh], 1 , 3 and 5 times) and 9 patients with prostate cancer frozen at -80 ℃ for different time (0 [i.e., fresh], 1, 2 and 4 weeks), and were statistically analyzed by Wilcoxon rank sum test for differences between the 2 groups. Results: The size distribution of exosomes extracted by the two methods was 30 to 150 nm by NTA, both of which were displayed as single peaks. The results of DLS showed that the potentials of exosome extracted by the two methods were negative values. The size of the exosomes extracted by the two methods was consistent observed under TEM namely the diameter distribution was 30 to 150 nm. western blot analysis confirmed that CD63 and Alix, the exosome labeling molecules, existed in the optimized method. The concentration of exosomes extracted from the 10 urine samples all reached 10 9 to 10 11 particles/mL. The contents of let-7c and PSA mRNA in exosomes decreased significantly after 5 freeze-thaw cycles, and the Z values were -1.79 and -1.73, respectively (P＜0.05). The RNA content of the exosomes remained stable after freezing at -80 ℃ for 1 month. Conclusion The optimized exosome extraction method could reduce greatly the cost under the premises of ensuring the concentration and quality of exosomes. The isolated exosomes may keep stable RNA content after freezing at -80 ℃ for a short time, but could not be frozen and thawed repeatedly for more than 5 times.