Objectives:Aimed to establish a rapid, high-throughput, automated method for determining the creatine kinase (CK-MM) isoform levels.Methods:Magnetic beads labeled with anti-CK-MM antibodies were combined with alkaline phosphatase-based chemiluminescence detection. Clinical and diagnostic performance validation of the assay was determined by analysis of 998 and 75 dried blood spot samples from healthy newborns and Duchenne muscular dystrophy (DMD) patients, respectively, and the CK activity was also determined. The blank and detection limits, cross-reactivity, recovery rate of the method, intra-and inter-assay coefficient, and the hook effect were evaluated.Results:Blank and detection limits were 17.4 and 39.3 ng/ml, respectively. Cross-reactivity toward CK-MB and CK-BB isoforms was 0.2% and 0.02%, respectively. Intra-and inter-assay coefficients of variation were<1%. Mean recovery was 100.32%, with no hook effect in CK-MM levels<50 000 ng/ml. Overall, the mean CK-MM concentrations in newborns and DMD patients were (27.05±0.97) and (3 720±300.5) ng/ml, respectively. A significant positive correlation between the dried blood spot detected CK-MM levels and total CK enzyme activity, evaluated in corresponding serum samples from the 75 DMD patients, was observed ( r=0.91, P<0.001), ?which is in good agreement with the clinical. Conclusions:An assay for rapid quantitative determination of CK-MM that meets clinical newborn screening requirements was established. It had a good value for application.

The fungus Xylaria sp. KYJ-15 was isolated from Illigera celebica. Based on the one strain many compounds (OSMAC) strategy, the strain was fermented on potato and rice solid media, respectively. As a result, two novel steroids, xylarsteroids A (1) and B (2), which are the first examples of C₂₈-steroid with an unusual β- and γ-lactone ring, respectively, along with two new dihydroisocoumarin glycosides, xylarglycosides A (3) and B (4), were identified. Their structures were elucidated by spectroscopic methods, X-ray diffraction and electronic circular dichroism (ECD) experiments. All isolated compounds were evaluated for cytotoxicity, DPPH radical scavenging activity, acetylcholinesterase inhibitory and antimicrobial effect. Compound 1 exhibited potent AChE inhibitory activity with an IC₅₀ value of 2.61 ± 0.05 μmol·L^-1. The β-lactone ring unit of 1 is critical for its AChE inhibitory activity. The finding was further confirmed through exploring the interaction of 1 with AChE by molecular docking. In addition, both compounds 1 and 2 exhibited obvious antibacterial activity against Bacillus subtilis with a minimum inhibitory concentration (MIC) of 2 μg·mL^-1. Compounds 3 and 4 exhibited antibacterial activities against Staphylococcus aureus with MICs of 4 and 2 μg·mL^-1, respectively, which also exhibited DPPH radical scavenging activity comparable to the positive control with IC₅₀ values of 9.2 ± 0.03 and 13.3 ± 0.01 μmol·L^-1, respectively.
Humans ; Acetylcholinesterase ; Molecular Docking Simulation ; Anti-Bacterial Agents ; Glycosides ; Lactones ; Pain

Metabolic reprogramming is a hallmark of cancer, including lung cancer. However, the exact underlying mechanism and therapeutic potential are largely unknown. Here we report that protein arginine methyltransferase 6 (PRMT6) is highly expressed in lung cancer and is required for cell metabolism, tumorigenicity, and cisplatin response of lung cancer. PRMT6 regulated the oxidative pentose phosphate pathway (PPP) flux and glycolysis pathway in human lung cancer by increasing the activity of 6-phospho-gluconate dehydrogenase (6PGD) and α-enolase (ENO1). Furthermore, PRMT6 methylated R324 of 6PGD to enhancing its activity; while methylation at R9 and R372 of ENO1 promotes formation of active ENO1 dimers and 2-phosphoglycerate (2-PG) binding to ENO1, respectively. Lastly, targeting PRMT6 blocked the oxidative PPP flux, glycolysis pathway, and tumor growth, as well as enhanced the anti-tumor effects of cisplatin in lung cancer. Together, this study demonstrates that PRMT6 acts as a post-translational modification (PTM) regulator of glucose metabolism, which leads to the pathogenesis of lung cancer. It was proven that the PRMT6-6PGD/ENO1 regulatory axis is an important determinant of carcinogenesis and may become a promising cancer therapeutic strategy.

BACKGROUND: Glutamine synthetase (GS) and arginase 1 (Arg1) are widely used pathological markers that discriminate hepatocellular carcinoma (HCC) from intrahepatic cholangiocarcinoma; however, their clinical significance in HCC remains unclear. METHODS: We retrospectively analyzed 431 HCC patients: 251 received hepatectomy alone, and the other 180 received sorafenib as adjuvant treatment after hepatectomy. Expression of GS and Arg1 in tumor specimens was evaluated using immunostaining. mRNA sequencing and immunostaining to detect progenitor markers (cytokeratin 19 [CK19] and epithelial cell adhesion molecule [EpCAM]) and mutant TP53 were also conducted. RESULTS: Up to 72.4% (312/431) of HCC tumors were GS positive (GS+). Of the patients receiving hepatectomy alone, GS negative (GS-) patients had significantly better overall survival (OS) and recurrence-free survival (RFS) than GS+ patients; negative expression of Arg1, which is exclusively expressed in GS- hepatocytes in the healthy liver, had a negative effect on prognosis. Of the patients with a high risk of recurrence who received additional sorafenib treatment, GS- patients tended to have better RFS than GS+ patients, regardless of the expression status of Arg1. GS+ HCC tumors exhibit many features of the established proliferation molecular stratification subtype, including poor differentiation, high alpha-fetoprotein levels, increased progenitor tumor cells, TP53 mutation, and upregulation of multiple tumor-related signaling pathways. CONCLUSIONS GS- HCC patients have a better prognosis and are more likely to benefit from sorafenib treatment after hepatectomy. Immunostaining of GS may provide a simple and applicable approach for HCC molecular stratification to predict prognosis and guide targeted therapy.
Humans ; Carcinoma, Hepatocellular/metabolism* ; Sorafenib/therapeutic use* ; Liver Neoplasms/metabolism* ; Glutamate-Ammonia Ligase/metabolism* ; Hepatectomy ; Retrospective Studies ; Prognosis ; Neoplasm Recurrence, Local/surgery*

Objective    To compare the short- and long-term efficacy of surgery and endoscopy in the treatment of early esophageal cancer by a systematic review and meta-analysis. Methods    We extracted data independently from The Cochrane Library, PubMed, EMbase, Web of Science for studies comparing surgery with endoscopy from 2010 to 2020. The primary outcomes including R0 resection rate, long-term overall survival (OS), disease-specific survival (DSS), major complications, recurrence, hospital stay and cost. Meta-analysis was performed using RevMan 5.3 and Engauge Digitizer was used to extract survival curves from relevant literature, and relevant data were calculated based on statistical methods. Results    A total of 17 studies involving 3 705 patients were included. It was found that patients in the surgery group had a higher R0 resection rate compared with the endoscopic group (OR=0.13, 95%CI 0.07 to 0.27, P<0.001, I2=6%). The total complications rate of resection of esophageal cancer was higher than that of the endoscopic group (OR=0.28, 95%CI 0.16 to 0.50, P<0.001, I2=68%). The length of hospitalization in the endoscopic group was obviously shorter than that in the surgery group (MD=–8.28, 95%CI –12.44 to –4.13, P<0.001, I2=96%). The distant recurrence rate (OR=0.58, 95%CI 0.24 to 1.41, P=0.230, I2=0%) and the local recurrence rate after resection (OR=1.74, 95%CI 0.66 to 4.59, P=0.260, I2=40%) in the endoscopic group was similar to those of the surgery group. There was no significant difference in 5 year-OS rate between the two groups (HR=0.86, 95%CI 0.67 to 1.11, P=0.25, I2=0%), which was subdivided into two groups: adenocarcinoma (HR=0.55, 95%CI 0.15 to 2.05, P=0.37, I2=0%) and squamous cell carcinoma (HR=0.68, 95%CI 0.46 to 1.01, P=0.06, I2=0%), showing that there was no difference between the two subgroups. There was no significant difference in the DSS rate (HR=0.72, 95%CI 0.49 to 1.05, P=0.090, I2=0%) between the two groups. The cost of the surgery group was significantly higher than that of the endoscopic group (MD=–12.97, 95%CI –18.02 to –7.92, P<0.001, I2=93%). Conclusion    The evidence shows that endotherapy may be an effective treatment for early esophageal neoplasm when considering the long-term outcomes whether it is squamous or adenocarcinoma, even though it is not as effective as surgery in the short-term efficacy.

Objective To examine the impact of vacuum sealing drainage on clinical outcomes of patients with post-sternotomy mediastinitis after cardiac surgery. Methods A systematic search were performed in Cochrane Library, Pubmed, Embase, China Biology Medicine(CBM), WanFang, VIP database. The quality of articles was critically appraised and data were extracted by 2 reviewers independently. Meta-analysis were conducted for the eligible researches. Results Fourteen cohort studies were inclued finally. Patients treated with VSD had significantly lower in-hospital mortalityand lower re-infection compared to those treated without VSD. While there had no significant defferences in length of ICU stay(days) and in-hospital stay (days) between VSD group and control group. Conclusions VSD therapy was associated with lower re-infection and in-hospital mortality than other conventional methods in patients with post-sternotomy mediastinitis after cardiac surgery and those results should be further tested in future research and practice.

Objective To evaluate the effect of case-PBL method and case-PBL method combined with SPSS software on the teaching of medical statistics for medical undergraduates. Methods Students from two classes major in experimental medicine from Nantong University Xingling College were selected as Experimental Class 1 ( using case-PBL teaching ) and Experimental Class 2 ( using case-PBL teaching combined with SPSS software). Each class attended lessons for five times. The teachers would send cases for each lesson one week in advance, and students would form into groups to access information, learn and discuss independently. Representatives from groups spoke in class, members would complement and discuss, while teachers offered guidance, review, analysis and summary. We shortened the discussion time in the class, and added instructions on the operation steps of SPSS software for case analysis and how to interpret the analysis result correctly in Experiment Class 2 . The Control Class received traditional teaching . Questionnaires, process assessment and final assessment were used to evaluate teaching effectiveness. SPSS 21.0 software was used to implement variance analysis and chi-square test for comparison between groups. Results The scores of the two experimental classes were higher than those of the control class in the five process assessments ( P<0 . 05 ) . Experimental Class 2 was the best in three section process assessments including those of t-test, the rank sum test and the straight-line correlation (P<0.05). However, there was no difference in the theoretical test scores among the three classes (P=0.670). 94％-100％of the students in the two experimental classes believed that the teaching effects of the case-PBL method and case-PBL method combined with SPSS software were better than the traditional teaching method (P<0.05) in such areas as stimulating interest in learning, improving learning efficiency, etc. Students in Experimental Class 2 were more prone to think these methods improved teaching effect. Conclusion The case-PBL teaching combined with the application of SPSS software will further help students to systematically grasp the statistical knowledge and cultivate statistical practice ability . However , neither methods will improve students' theoretical test scores.

Objective To compare the clinical value of XELOX and FOLFOX6 in the treatment of elderly patients with colorectal cancer. Methods Clinical data of 68 elderly patients(aged>60 years)with colorectal cancer were retrospectively analyzed.The patients were divided into XELOX group(n =32)and FOLFOX6 group(n =36) according to the chemotherapy regimen.The response evaluation criteria in solid tumors,progression free survival, 2 -year mortality,chemotherapy related adverse reactions and health related quality of life in the two groups were compared.Results Compared with the FOLFOX6 group,the neurotoxicity incidence in the XELOX group was signifi-cantly lower (3.13% vs.22.22%,χ2 =5.380,P =0.020).The neutrophils reduced incidence was significantly lower(9.38% vs.30.56%,χ2 =4.649,P =0.039).The hand foot syndrome complications significantly increased (34.38% vs.8.33%,χ2 =7.027,P =0.008).There was no significant difference in the incidence of nausea,vomi-ting,diarrhea,oral mucosal ulcer,thrombocytopenia and elevation of transaminase in the two groups(χ2 =0.001, 0.000,0.008,2.102,1.071,1.851,all P >0.05).The quality of health related quality of life was significantly higher in the XELOX group than in the FOLFOX6 group at 6 months after the operation[(67.94 ±12.49)points vs. (57.39 ±13.59)points,t =3.198,P =0.015].There were no significant differences in the curative effect grade at 6 months after the operation,2 -year mortality rate and progression free survival time between the two groups(χ2 =0.949,0.008,t =0.573,all P >0.05).Conclusion Although XELOX increases the incidence of hand foot syn-drome in elderly patients with colorectal cancer,but it significantly reduces the incidence of neurotoxicity and neutro-phil,and improves the quality of life of patients after surgery.

Objective To summarize the experience ofmultidisciplinary team (MDT) in diagnosis and treatment of complicated and refractory thyroid tumors.Methods A retrospective review was performed on clinical data of 46 cases with complicated and refractory large thyroid tumors admitted to our hospital from Jan.2010 to Dec.2016.There were 23 cases in MDT group and 23 cases in the control group,respectively.The MDT group received diagnosis and treatment provided by multidisciplinary team during perioperative period whereas the control group received conventional surgery.Results Short-term complications such as trouble breathing and thyroid crisis were not observed in 46 patients after surgery.In the control group,the mean durations were (52±11.5)minutes for anesthesia,(159±38.1) minutes for surgery and (11 ±3.5) days for hospital stay,respectively.After surgery,bleeding occurred in 5 cases,hoarseness in 5 cases,irritating cough when drinking in 7 cases,transient hypocalcemia in 8 cases,permanent hypocalcemia in 6 cases,and neck tracheotomy due to tracheomalacia during surgery in 2 cases.In MDT group,the mean durations were (37±8.5) minutes for anesthesia,(134±28.5) minutes for surgery and (7±1.5) days,respectively.After surgery,bleeding occurred in 0 case,hoarseness in 0 case,irritating cough when drinking in 1 case,transient hypocalcemia in 2 cases,permanent hypocalcemia in 0 case,and neck tracheotomy due to tracheomalacia during surgery in 4 cases.Conclusion Application of multidisciplinary team in diagnosis and treatment of complicated and refractory thyroid tumors can reduce duration of preoperative endotracheal anesthesia as well as surgery,decrease postoperative complications,shorten duration of hospitalization and improve life quality after surgery.

Objective To explore the effects of different vitamin A(VA) levels on thyroid cells apoptosis and its gene expression of mice taking excessive iodine. Methods Kunming mice were randomly divided into 6 groups according to body weight 3 weeks after born: normal control(NI) group, high iodine(HI) group, low vitamin (LVA) group, high iodine plus low vitamin A(HI+LVA) group, high iodine plus vitamin A1 (HI+VA1) group, high iodine plus vitamin A2(HI+VA2) group. The VA was given in food(4000,4000,0,0,8000,16 000 U/kg), and the iodine was given as potassium iodate in water (I-:50,3000,50,3000,3000,3000 μg/L). The apoptosis was tested using in situ end labehng(TUNEL) method. Reverse transcription polymerase chain reaction (RT-PCR) were used to measure the level of mRNA of apoptosis gene(Fas, FasL, Bcl-2) in tissues. Results Apoptotic index measured by TUNEL method was rising along with the mice age. Compared to NI group[(14.09±5.68)%], apoptotic index was significantly increased in HI[(20.91±9.57)%], HI+LVA[(20.29±9.90)%]and HI+VA2 [(19.51±8.25)%]groups in the three months(P < 0.05). Compared to NI group[(16.80±9.90)%], apoptotic index was significantly increased(P < 0.05) in HI[(23.22±8.58)%],LVA[(22.56±6.17)%],HI+LVA [(25.99±9.62)%],HI+VA1 [(21.65±7.74)%]groups in the six months. Compared with the NI group(Fas: 1.29±0.25,1.27±0.26; FasL: 1.60±0.13,1.65±0.13), the mRNA levels of Fas and FasL in HI group(Fas: 1.57±0.36,1.49±0.35; FasL: 1.85±0.46,1.84±0.32) were increased, but the differences were not remarkable(P > 0.05) in the three and six months. Compared with the HI group, the mRNA levels of Fas in HI+ VA1, HI+VA2(1.33±0.35, 1.30±0.26) groups were decreased to the level in NI group in the six months. The mRNA levels of Fas and FasL were not different (P > 0.05) between HI+LVA(I.60±0.27,1.88±0.46) and HI groups in the three months. The mRNA levels of Bcl-2 were not remarkably differences in the three months (1.05±0.19,0.96±0.33,0.95±0.26,1.18±0.27,1.10±0.19,0.98±0.36, all P > 0.05), and in the six months (1.35±0.28,1.60±0.25,1.48±0.18,1.71±0.26,1.66±0.29,1.56±0.35, all P > 0.05). Conclusions Excessive iodine can cause thyroid cells apoptosis in mice. Supplementation of suitable amount of VA can regulate the levels of the apoptosis-related genes expression, and partly antagonize the apoptosis caused by high iodine.