Objective To study the value of serum S-100B protein in the diagnosis of cerebral radi-ation injuries in patients with brain malignant tumor. Methods Serum S-100B protein level was deteetod by enzyme-linked immunosorbent assay in 56 patients with brain malignant tumor before, during and after radio-therapy. Effects of dose and method of radiotherapy, peritumoral edema degree and Karnofsky performance status on serum S-100B level were studied. Results The levels of serum S-100B protein in the patients be-fore radiotherapy and control group were 0. 039μg/L and 0.044 μg/L ( t = 1.48 ,P =0. 186). The levels of serum S-100B protein before, in the middle of (30-40 Gy) and after (60 -70 Gy) radiotherapy were 0.044 μ/L, 0.049 μ/L and 0.079 μg/L, respectively ( F = 67.26, P = 0.000). The differences after ra-diotherapy were also significant among patients with three methods of radiotherapy (F = 20.32, P = 0.000), different degree of pefitumoral edema ( F = 12.94, P =0. 000 ) and Karnofsky perforrnanee status ( t = 2.71, P =0.007). Conclusions High level of serum S-100B protein is associated with cerebral radiation injuries in patients with brain malignant tumor, which is influenced by the dose and method of radiotherapy, Karnof-sky performance stares and degree of peritumoral edema. High level of serum S-100B protein may serve as an early predictor of cerebral radiation injury.

Objective At present, the methods of separating and identifying nasopharyngeal cancer stem cells are not yet mature.This study was to explore the methods of culturing nasopharyngeal cancer stem cell microspheres and identify the cancerous stem cell biological features of CNE-2 cell microspheres. Methods We conducted suspension culture of human nasopharyngeal cancer CNE2 cells and C666-1 cells in serum-free medium ( SFM) containing growth factors.Then we measured the proportion of CD133 +cells in CNE2 monolayer ( CNE2-MN) and CNE2 microsphere cells ( CNE2-SC) by flow cytometry, determined their in vitro invasiveness through Transwell chamber experiments, and detected their in vivo tumorigenicity via nude mouse experiments.We ob-served the differentiation potency of the CNE2-SCs in the adherent-cultured serum-containing medium and detected the expressions of the cancer stem cell-related genes Bmi-1, Oct4, and Twist1 in CNE2-MN and CNE2-SCs by flow cytometry and RT-PCR analysis. Results In the special blend of SFM, both of the cell lines can form microspheres that can be stably transferred.Fresh SFM prepara-tion, substituting cell separation agent Accutase for pancreatic enzyme for transfer, and maintaining the state of cell suspension contrib-uted to the formation and proliferation of microspheres.Adherent culture with serum-containing medium induced the differentiation of CNE2-MN cells, which exhibited no significant difference from the CNE2 microsphere cells.The CD133 +cells accounted for 98.79%in the CNE2 microspheres, significantly higher than 0.98%in the CNE2 cells (P<0.01).Compared with the CNE2-MN cells, the CNE2-SCs showed highly increased expressions of Bmi-1, Oct4, and Twist1 (P<0.01).The numbers of membrane-penetrating cells in the CNE2-SCs and CNE2-MN cells were 122 ±6 and 36 ±7 per visual field, the former with a stronger invasive ability than the latter genesis of 1 ×106 CNE2-SCs vs that of the same number of CNE2 -MNcells was (2.332 ±0.549) cm 3 sv (0.669 ±0 .278) cm3 ( P<0.01). Conclusion Using suspension culture with a specially prepared SFM, nasopharyngeal cancer CNE2 microsphere cells can be obtained, in which there are large numbers of cancer stem cells.This culture method may provide a base for further studies of naso-pharyngeal cancer stem cells.

A method was developed for determination of 12 kinds of phosphate compounds in water and sediment by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) coupled with solid phase extraction (SPE) and ultrasonic extraction.The water samples were concentrated by HLB solid-phase extraction (SPE) column and eluted twice with ethyl acetate, ultrasonic solvent extraction for sediment samples and then repeated the operation of water samples after diluted with deionized water.The sample were separated on a ZORBAX Eclipse Plus C18 (150 mm × 2.1 mm, 3.5 μm) column by a gradient elution with 0.2% formic acid aqueous solution and methanol as the mobile phase.Ion mode analysis was monitored by high performance liquid chromatography mass spectrometer (MRM).The target compounds were quantified by external standard method.At the spiked levels (0.05, 0.1 and 0.5 μg/L), the average recoveries of 12 kinds of OPEs in water samples ranged from 66.4% to 115%, except for TMP (28.5%-47.8%) and TEHP (22.4%-73.8%).The relative standard deviation RSD (n=3) was 0.5%-9.09%, and the method quantification (MOQ) was 0.001-0.05 μg/L, However at the spiked levels of 5, 10 and 50 μg/kg, the average recoveries of 12 kinds of OPEs in sediment samples ranged from 65.4% to 120.0%, except for TMP (35.7%-44.9%) and TCEP (31.2%-48.9%).The relative standard deviation RSD (n=3) was 0.01%-9.54%, and the MOQ for sediment was 0.02-2.0 μg/kg dw.Based on the above methods, the detection and analysis of the targets in the water and sediments samples of Taihu Lake were carried out.The results showed that the concentrations of ΣOPEs were 0.1-1.7 μg/L and 8.1-420 μg/(kg dw), respectively.

AIM: To illuminate the effect of SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily b, member 1 (SMARCB1) in early diagnosis and prognosis of hepatocellular carcinoma (HCC) by determining the clinical expression of SMARCB1 in HCC tissue and benign liver tissue.METHODS: The specific target gene SMARCB1 was selected from these genes by using The Cancer Genome Atlas (TCGA).SMARCB1 expression in HCC tissue and benign liver tissue was measured by immunohistochemistry.Further statistical analysis of TCGA was performed to illuminate the role of SMARCB1 on HCC occurrence and progression.RESULTS: Compared with the benign liver tissue, immunohistochemical staining showed that SMARCB1 expression was significantly up-regulated in the HCC tissue (P<0.01).In addition, SMARCB1 expression was significantly associated with advanced tumor stage (P<0.05).The relation between SMARCB1 expression at mRNA level and clinical prognosis was analyzed.The results indicated that high SMARCB1 expression was an independent prognostic factor for HCC (P<0.05).CONCLUSION: SMARCB1 may play a part as a carcinogenic gene in tumorigenesis.We can distinguish primary HCC samples from non-malignant samples according to its different clinical expression.High SMARCB1 expression probably predicts poor outcome in HCC patients.

Objective There is still a lack of effective clinical prognostic factors for predicting outcomes and guiding treatments in extranodal nasal-type NK/T-cell lymphoma (NKTCL).This study was aimed to investigate the clinical features and prognostic role of primary tumor burden (PTB).Methods A total of 1383 patients were recruited from ten hospitals, including 947 stage Ⅰ patients (68.5%), 326 stage Ⅱ patients (23.6%), and 110 stage Ⅲ-IV patients (8.0%).There were 751 patients (54.3%) presenting with high PTB (H-PTB).The Kaplan-Meier method was used to calculate survival rates, and the log-rank test was conducted for survival difference analysis.Meanwhile, a multivariate analysis was performed using the Cox regression model.Results H-PTB was associated with high invasive potential, high frequency of B symptoms, advanced stage, regional lymph node involvement, lactate dehydrogenase elevation, and poor performance status.The patients with H-PTB had significantly lower 5-year overall survival (OS) and progression-free survival (PFS) rates than those with low PTB (L-PTB)(OS:50.2% vs.72.1%, P=0.000;PFS:41.8% vs.62.5, P=0.000).PTB was an independent prognostic factor for both OS (HR=1.851) and PFS (HR=1.755) according to the Cox multivariate analysis.Moreover, H-PTB was associated with significantly lower locoregional control (LRC) in early-stage NKTCL, and the 5-year LRC rate was 71.6% in patients with H-PTB and 84.3% in those with L-PTB (P=0.000).Conclusions H-PTB is associated with multiple adverse clinical features in NKTCL, and it is an independent indicator for poor outcomes and LRC.H-PTB can be used as a reliable indicator for risk stratification and treatment decision.

Objective To evaluate the prognostic value of anemia in patients diagnosed with extranodal nasal-type natural killer (NK)/T cell lymphoma (NKTCL).Methods Clinical data of 1 225 NKTCL patients receiving the first course of treatment from 10 medical institutions in China were retrospectively analyzed.According to the diagnostic criteria in China,anemia was defined as the hemoglobin (Hb) level＜ 120 g/L for the male and＜ 110 g/L for the female from the sea-level area.The severity of anemia was classified into the extremely severe anemia (Hb ≤ 30 g/L),severe anemia (31-60 g/L),moderate anemia (61-90 g/L) and mild anemia (＞90 g/L).Results Among 1 225 patients,199(16.2％) were complicated with anemia,who had more adverse prognostic factors compared with their counterparts without anemia.Among NKTCL patients with anemia,the proportion of patients with stage Ⅱ-ⅣV,a median age＞ anemia,Eastern Cooperative Oncology Group (ECOG) score of 2-4 and NK/T-cell lymphoma prognostic index (NKTCLPI) ≥ 2 was relatively high.Patients with anemia obtained worse clinical prognosis than those without anemia.The 5-year overall survival (OS) and progression-free survival (PFS) in NKTCL patients with anemia were calculated as 49.4％ and 35.4％,significantly lower compared with 63.3％ and 56.0％ in their counterparts without anemia (both P＜0.01).Single factor analysis demonstrated that anemia,age,ECOG score,group B symptom,lactate dehydrogenase,primary tumor site,primary tumor invasion and staging were the prognostic factors of OS and PFS.Multivariate analysis revealed that anemia was still the independent prognostic factor.Conclusions Anemia is not common in patients with NKTCL and these patients obtain poor clinical prognosis.Anemia is an independent prognostic factor for patients with NKTCL.

Objective To investigate the role of ceramide in radiation induced bystander effect (RIBE) in vivo by irradiating Caenorhabditis elegans with proton microbeam.Methods The posterior pharynx of wide-type N2 and genetically mutated L4-staged worms was irradiated with 2 000 particles,then germ cell apoptosis and gene expression were analyzed.Results Point-fixed radiation to posterior pharynx of N2 worms significantly increased the number of apoptotic germ cells (t =9.007,P＜0.05),but not in the worms with deletion mutants of ceramide synthase (CerS) genes (hyl-1 and lagr-1) (P＞0.05).Realtime quantitative PCR assay indicated that in both N2 and lagr-1 (gk327);hyl-1 (ok976) worms,the genes of egl-1 and ced-13 in core apoptosis pathway were up-regulated after radiation to posterior pharynx,without statistical significance between these two strains (P＞0.05).The gene expression levels of hyl-1 and lagr-1 were increased in both N2 and hus-1 (op241) worms after radiation with no statistical significant difference between two strains (P＞0.05).Furthermore,radiation-induced germ cell apoptosis was increased in abl-1 (ok171) worms (t=13.241,P＜0.05),rather than in lagr-1(gk327);hyl-1(ok976);abl-1(ok171) worms (t=13.462,P＜0.05).Conclusions Ceramide is required for RIBE on germ cell apoptosis in C.elegans,and it might play function together with egl-1 and ced-13 in core apoptosis pathway and HUS-1 in DNA damage response pathway.But ceramide had antagonistic relationship with anti-apoptotic protein abl-1 in germ cell apoptosis.

Polyhydroxyalkanoates (PHAs) have obtained much attention in biomaterial fields due to their similar physicochemical properties to those of the petroleum-derived plastics. Poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)] is one member of the PHAs family, and has better toughness and transparency compared to existing polylactic acid (PLA) and poly[(R)-3-hydroxybutyrate] [P(3HB)]. First, we confirmed the one-step biosynthesis of P(LA-co-3HB) with the lactate fraction of 23.8 mol% by introducing P(3HB-co-LA) production module into Escherichia coli MG1655. Then, the lactate fraction was increased to 37.2 mol% in the dld deficient strain WXJ01-03. The genes encoding the thioesterases, ydiI and yciA, were further knocked out, and the lactate fraction in the P(3HB-co-LA) was improved to 42.3 mol% and 41.1 mol% respectively. Strain WXJ03-03 with dld, ydiI and yciA deficient was used for the production of the LA-enriched polymer, and the lactate fraction was improved to 46.1 mol%. Notably, the lactate fraction in P(3HB-co-LA) from xylose was remarkably higher than from glucose, indicating xylose as a potent carbon source for P(3HB-co-LA) production. Therefore, the deficiency of thioesterase may be considered as an effective strategy to improve the lactate fraction in P(3HB-co-LA) in xylose fermentation.
Escherichia coli/genetics* ; Hydroxybutyrates ; Lactic Acid ; Polyesters ; Polyhydroxyalkanoates ; Xylose

OBJECTIVE To stu dy the effects of different processing me thods on the contents of the pharmacodynamic index components in Citrus aurantium and their antioxidant activity. METHODS According to the general principles of 2020 edition of Chinese Pharmacopoeia （volume Ⅳ） and the relevant processing methods in 2015 edition of Processing Specifications of Traditional Chinese Medicine in Zhejiang Province ，the samples of C. aurantium were prepared by steaming with water ，boiling with water ，stir-frying with vinegar ，stir-frying with wine ，stir-frying with bran ，processing with bran and processing with honey. The contents of moisture and ash in different products of C. aurantium were detected. The contents of naringin and neohesperidin in different products of C. aurantium were determined by high performance liquid chromatography. The antioxidant activity of different products was investigated by DPPH and ABTS + radical scavenging experiments and the total reducing power test. RESULTS The contents of moisture ，ash，naringin and neohesperidin were in line with the relevant requirements in 2015 edition of Processing Specifications of Traditional Chinese Medicine in Zhejiang Province . The content of naringin in descending order was as follow ： unprocessed sample ＞sample processed with honey ＞sample processed with bran ＞sample boiled with water ＞sample stir-fried with vinegar＞sample stir-fried with wine ＞sample stir-fried with bran ＞sample steamed with water. The content of neohesperidin in descending order was as follow ：unprocessed sample ＞sample boiled with water ＞sample processed with bran ＞sample processed with honey ＞sample stir-fried with vinegar ＞sample steamed with water ＞sample stir-fried with wine ＞sample stir-fried with bran. The samples after boiling with water ，processing with bran ，and stir-fried with bran had better DPPH radicals scavenging ability （IC50 were 7.49，8.37 and 10.22 mg/mL，respectively）. The samples after boiling with water ，steaming with water ，and processed with bran had better ABTS + radicals scavenging ability （IC50 were 1.76，2.03 and 2.72 mg/mL，respectively）. In addition ， compared with sample stir-fried with wine and processed with 发。E-mail：wanglu1286@163.com honey，unprocessed sample and other processed products of C.aurantium had bet ter total reducing ability. CONCLUSIONS After processing ，the contents of the main pharmacodynamic index components in C. aurantium have been reduced ，but they were also in line with the relevant requirements in 2015 edition of Processing Specifications of Traditional Chinese Medicine in Zhejiang Province . The antioxidant ability of some processed products has been enhanced.