Background and Aims:Accurate early pathogen diagnosis is a breakthrough for improving the prognosis of infectious pancreatic necrosis(IPN)patients.However,there is currently a lack of efficient methods for early identification of IPN in clinical settings.This study was performed to assess the application value of next-generation sequencing technology based on metagenomic capture(MetaCAP)in the pathogen diagnosis of IPN. Methods:A prospective study was conducted on 29 patients suspected of having acute necrotizing pancreatitis at Xiangya Hospital of Central South University between January and July 2024.Blood samples were tested using MetaCAP and conventional pathogen culture.The results of peritoneal fluid pathogen culture were used as the gold standard to compare the diagnostic efficacy of the two methods. Results:Due to three cases lacking peritoneal fluid culture results,a total of 26 cases were included in the final analysis.The overall mortality rate was 23.1％(6/26).During hospitalization,9 cases(34.6％)were diagnosed with IPN.The sensitivity and negative predictive value of MetaCAP for diagnosing IPN were significantly higher than those of conventional pathogen culture(77.8％vs.11.1％,P=0.031;86.7％vs.65.2％,P=0.032),while the differences in specificity(76.5％vs.88.2％,P=0.689)and positive predictive value(63.6％vs.33.3％,P=0.347)between the two methods were not statistically significant.The average detection time for MetaCAP was 33(20-49)h,while microbial culture took 125(45-142)h,with a significant difference(P＜0.001).The average cost for blood MetaCAP testing was 2 500 yuan per case,but it accounted for only 1.19％of the average hospitalization cost. Conclusion:MetaCAP has significant value in the early pathogen diagnosis of IPN,with a shorter detection time,good testing efficacy,and health-economic value,demonstrating a promising clinical application prospect.

Different types of tumors or common anti-tumor treatment measures can lead to cancer-re-lated cognitive impairment (CRCI) appearance in pediatric patients.Based on the theory of unpleasant symp-toms (TOUS),this paper summarizes a total of 20 key influencing factors in 3 levels:the physiological level includes the disease type,treatment method,treatment-related complications,age,gender,etc.;the psychologi-cal level encompasses fatigue,anxiety,depression,and sleep disturbances,etc.;the background level involves the family environment and socioeconomic status,parental education level,physical activity,etc.This paper e-lucidates these influencing factors to provide reference for further research in this field.

Objective:To investigate the application value of metagenomic next-genera-tion sequencing (mNGS) in pathogenic diagnosis of suspected infected severe acute pancreatitis (SAP).Methods:The prospective study was conducted. The clinical data of 25 patients with suspected infected SAP who were admitted to the Xiangya Hospital of Central South University from May to September 2023 were collected. Upper limb venous blood samples of all the patients were collected for both of mNGS and routine pathogen microbial culture. Observation indicators: (1) grouping situations of the enrolled patients; (2) comparison of the diagnostic efficiency of mNGS and routine pathogen microbial culture; (3) results of peripheral blood pathogen microbial testing and peri-pancreatic effusion microbial culture; (4) testing time and cost. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3). Count data were expressed as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Results:(1) Grouping situations of the enrolled patients. A total of 25 patients were selected for eligibility. There were 18 males and 7 females, aged 48(40,59)years. The duration of hospital stay of 25 patients was 30(20,50)days. The etiologies of 25 patients included 14 cases of hyperlipidemic pancreatitis, 8 cases of biliary pancreatitis, 1 case of alcohol-induced acute pancreatitis, and 2 cases of pancreatitis caused by other causes. Of the 25 patients, there were 17 cases with infected pancreatic necrosis (IPN) including 7 cases of death, and 8 cases with sterile pancreatic necrosis including no death. (2) Comparison of the diagnostic efficiency of mNGS and routine pathogen microbial culture. The positive rates of mNGS and routine pathogen microbial culture in diagnosis of suspected infected SAP were 72.0%(18/25) and 32.0%(8/25), respectively, showing a significant difference between them ( χ2=8.01, P<0.05). The sensitivity and negative predic-tive value of mNGS and routine pathogen microbial culture in diagnosis of IPN were 94.1%(16/17), 35.3%(6/17) and 85.7%(6/7), 35.3%(6/17), showing significant differences between them ( χ2=12.88, 5.04, P<0.05). The specificity and positive predictive value of mNGS and routine pathogen microbial culture in diagnosis of IPN were 75.0%(6/8), 75.0%(6/8) and 88.9%(16/18), 75.0%(6/8), showing no significant difference between them ( χ2=0, 0.82, P>0.05). (3) Results of peripheral blood pathogen microbial testing and peripancreatic effusion microbial culture. Of the 17 patients with IPN, 36 strains of pathogenic bacteria were detected by mNGS, and 6 strains were detected by routine pathogen microbial culture. There were 16 of 17 patients with IPN showing positive mNGS pathogenic testing, of which 13 cases were consistent with the pathogenic testing results of peri-pancreatic effusion microbial culture, showing a consistency rate of 76.5%(13/17). There were 6 pati-ents with IPN showing positive routine pathogen microbial culture, with a consistency rate of 35.3%(6/17) to peripancreatic effusion microbial culture. (4) Testing time and cost. Testing time of mNGS and routine pathogen microbial culture were (43±17)hours and (111±36)hours, showing a signifi-cant difference between them ( t=9.31, P<0.05). Testing cost of mNGS was (2 267±0)yuan/case, accoun-ting for 1.7% of the hospitalization expenses of (133 759±120 744)yuan/case. Testing cost of routine pathogen microbial culture was (240±0)yuan/case, accounting of 0.2% of the hospitalization expenses. Conclusion:mNGS has important value for early pathogenic diagnosis of suspected infected SAP, and has a high timeliness.

Objective:To describe the distributions of demographic and clinic pathological characteristics and relations with survival on female breast cancer patients in Guangzhou from 2008 to 2017.Methods:The baseline information of the subjects was obtained from the Guangzhou cancer registry and the outcomes were from the Cancer Follow-up System of Guangzhou. Kaplan-Meier was used to calculate the 1-, 3-, 5-year overall survival rates. Univariate and multivariate Cox proportional hazards regression models were used to identify the factors related to the overall survival.Results:Among the 12 465 breast cancer patients recruited in the study, the average age at diagnosis was 53.9 years old, with those aged 45 to 54 making up the largest proportion (43.9 %). Only 15.6 % of the patients had college or above degrees. Patients with normal BMI accounted for 78.2 %. Most of the patients (90.0 %) had received surgical treatment. Invasive ductal carcinoma appeared the most common histologic type, accounting for 82.3 %. Among the 2 640 patients diagnosed in the four large hospitals, clinical stages 0-Ⅰ, Ⅱ, Ⅲ and Ⅳ accounted for 35.0 %, 44.8 %, 17.2 % and 3.0 %, respectively. The proportions of ER-positive, PR-positive and HER-2 positive breast cancer were 79.5 %, 70.8 %, and 19.2 %, respectively. In terms of subtypes, Luminal B was the most common one, accounted for 53.3 %. The 1-, 3- and 5-year overall survival rates were 99.0 %, 95.3 % and 92.1 %, respectively. Results from the multivariate analysis indicated that factors as: age over 55 years old at diagnosis, advanced TNM stage, ER negative, PR negative, Luminal B subtype and triple-negative subtype were associated with poorer prognosis. Conclusions:Compared with the previous hospital-based studies in China, this population-based study revealed that the proportions of patients with advanced age, early clinical stage or ER positive breast cancer were relatively high and the overall survival rate for breast cancer was higher than that in the previous studies. Relationships between characteristics and prognosis of breast cancer were consistent with the previous findings.

Antiviral Agents ; chemistry ; metabolism ; Crystallography, X-Ray ; Ligands ; Protein Binding ; Protein Structure, Tertiary ; RNA Helicases ; chemistry ; classification ; metabolism ; Static Electricity ; Viral Proteins ; chemistry ; classification ; metabolism ; Zika Virus ; enzymology

The recent explosive outbreak of Zika virus (ZIKV) infection has been reported in South and Central America and the Caribbean. Neonatal microcephaly associated with ZIKV infection has already caused a public health emergency of international concern. No specific vaccines or drugs are currently available to treat ZIKV infection. The ZIKV helicase, which plays a pivotal role in viral RNA replication, is an attractive target for therapy. We determined the crystal structures of ZIKV helicase-ATP-Mn(2+) and ZIKV helicase-RNA. This is the first structure of any flavivirus helicase bound to ATP. Comparisons with related flavivirus helicases have shown that although the critical P-loop in the active site has variable conformations among different species, it adopts an identical mode to recognize ATP/Mn(2+). The structure of ZIKV helicase-RNA has revealed that upon RNA binding, rotations of the motor domains can cause significant conformational changes. Strikingly, although ZIKV and dengue virus (DENV) apo-helicases share conserved residues for RNA binding, their different manners of motor domain rotations result in distinct individual modes for RNA recognition. It suggests that flavivirus helicases could have evolved a conserved engine to convert chemical energy from nucleoside triphosphate to mechanical energy for RNA unwinding, but different motor domain rotations result in variable RNA recognition modes to adapt to individual viral replication.
Crystallography, X-Ray ; Protein Domains ; RNA Helicases ; chemistry ; RNA, Viral ; chemistry ; Viral Proteins ; chemistry ; Zika Virus ; enzymology

Objective To study the effect of pioglitazone(PIO) on AdipoR1 and cholesterol ester(CE) in foam cells derived from THP-1-derived macrophages.Methods THP-1-derived macrophages were incubated with increasing concentrations of PIO for 24 hours.After co-cultured with low density lipoprotein(LDL),the accumulation of cholesterol in macrophages was measured by fluorescence spectrophotometric method.The lipid peroxide within cells was detected by TBARS method,the foam cells were observed by oil red staining.AdipoR1 levels were determined by Western blot.Results Compared with the ox-LDL group (0 μmol/L),oil red O-positive cells of the PIO protective groups were greatly reduced.TC,CE,MDA of the PIO protective groups were also obviously decreased.TC (53.6 ± 1.2) μg/mg,CE (30.2 ± 3.6) μg/mg,MDA (3.42 ± 0.06) μg/mg of 5 μμ mol/L PIO group were lower than those of 0μmol/L PIO group[(98.2 ± 3.5),(65.5 ± 6.5),(8.50 ± 1.21)] μg/mg (P ＜ 0.05).TC (25.6 ± 1.8) μg/mg,CE (22.5 ± 4.5) μg/mg,MDA (1.90 ± 0.42) μg/mg of 50 μmol/L PIO group.TC (16.8 ± 2.2) μg/mg,CE(5.9 ± 1.4) μg/mg,MDA (0.65 ± 0.05) μg/mg of 100μmol/L PIO group.Concomitantly,PIO significantly increased AdipoR1 protein expresion,AdipoR1 of 5μmol/L PIO group(0.06±0.05) was higher than that of 0μmol/L PIO group(0.03 ±0.07).AdipoR1 of 50μmol/L PIO group(0.11 ±0.07) was higher than that of 5μmol/L PIO group (0.06 ± 0.05).AdipoR1 of 100 μmol/L PIO group (0.40 ± 0.05) was obviously higher than that of 50 μ mol/L PIO group (0.11 ± 0.07).Conclusion PIO inhibited THP-1-derived formation by up-regulation the expression of AdipoR1,which may play an important role in the development and progression of atherosclerosis.

BACKGROUNDnm23-H1 gene is a well-known tumor metastasis suppression gene. Our previous study has found that transfection of wild type nm23-H1 gene can significantly downregulate the ERK1/2 activity of human high-metastatic large cell lung cancer cell line L9981. The aim of this study is to investigate the influence of nm23-H1 and exogenous ERK1/2 pathway inhibitor U0126 on the extracellular signal-regulated kinase (ERK1/2) of human high-metastatic large cell lung cancer cell line L9981 and its malignant biological behaviors.

METHODSThe expressive levels of total-ERK1/2, dually phosphorylated ERK1/2 and ERK1/2 relative activity of the human high-metastatic large cell lung cancer cell lines, L9981 (parent cell line with nm23-H1 gene hetero-deletion), L9981-nm23-H1 (transfected with nm23-H1 gene ) and L9981-PLXSN (transfected with vector) were detected by Western blot and immunoprecipitation technique after treating with U0126 (40μmol/L for 20 minutes). The in vitro proliferative and invasive abilities among the above three lung cancer cell lines were determined by MTT and improved Boyden chamber methods.

RESULTSThe phosphorylated ERK1/2 expression level and relative activity in L9981-nm23-H1 lung cancer cell line were remarkably lower than those in L9981 and L9981-PLXSN lung cancer cell lines after being treated with U0126 (P < 0.01), but there was no significant difference between L9981 and L9981-PLXSN lung cancer cell lines. No significant difference of total ERK1/2 expression level was observed among the three lung cancer cell lines (P > 0.05) after being treated with U0126. The in vitro proliferation and invasion of L9981-nm-23H1 lung cancer cell line were remarkably lower than those of L9981 and L9981-PLXSN lung cancer cell lines (P < 0.01 ), but no significant difference was found between L9981 and L9981-PLXSN lung cancer cell lines (P > 0.05 ); U0126 could significantly down-regulate the in vitro proliferation and invasion of L9981 lung cancer cell line (P < 0.01).

CONCLUSIONSBlocking the activity of ERK1/2 in L9981 lung cancer cell line and transfecting the nm23-H1 gene into the L9981 lung cancer cell line may produce similar cell biological behavior changes, namely the significant reduction of in vitro proliferation and invasion of L9981 lung cancer cell line. These results indicate that the molecular mechanism which nm23-H1 gene reverses invasion and proliferation of the human high-metastatic large cell lung cancer cell line may be related to its effects of down-regulating the activity of the key kinase ERK1/2 of Ras-to-MAPK signal transduction pathway.

nm23-H1 is a proven tumor metastasis suppressive gene, tumor metastasis phenotype could be reversed by transfected nm23-H1 cDNA. This study was conducted to transfect nm23-H1 cDNA into L9981 cells and to explore the function of nm23-H1 in reversing the malignant phenotype of L9981 cells. The plasmid of pLXSN-nm23-H1-EGFP was constructed by gene clone technique, and the transfected nm23-H1 cDNA cell lines of L9981-nm23-H1 was established. The protein expression of nm23-H1 was detected by Western blot. The biologic features of L9981-nm23-H1 cells were studied in vitro and in vivo. The results showed that the fusion protein of nm23-H1-EGFP was stable, continuous and expressed with high efficiency in L9981-nm23-H1 cells. The cell proliferation, colon formation and invasive ability are significantly lowered in L9981 cells transfected nm23-H1 cDNA (P < 0.01); the tumorgenesis and the lung metastasis incidence was lower in tranfected nm23-H1 cells than in L9981 and L9981-Plxsn in nude mice (P < 0.01); the rate for inhibiting tumorgenesis of nm23 -H1 was 82.56%. These data suggest that the malignant phenotype could be reversed by wild nm23-H1 gene in L9981 cells.
Animals ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Lung Neoplasms ; pathology ; Mice ; Mice, Nude ; NM23 Nucleoside Diphosphate Kinases ; genetics ; Neoplasm Metastasis ; Neoplasm Transplantation ; Recombinant Fusion Proteins ; genetics ; Transfection ; Tumor Suppressor Proteins ; genetics

BACKGROUNDIt has been proved that tumor development and metastasis are dependent on angiogenesis. Suppression of tumor angiogenesis can inhibit tumor growth and metastasis. Collagen X VIII/endostatin is one of the most effective inhibitors of angiogenesis at present. The aim of this study is to study the relationship between transcription expression of endostatin mRNA and clinical and pathophysiological characteristics in non-small cell lung cancer (NSCLC).

METHODSThe transcription expression of endostatin mRNA was detected in 46 lung cancer tissues and paracancerous lung tissues, 14 benign pulmonary lesion tissues as control by RT-PCR method.

RESULTS(1)The transcription expression of endostatin mRNA in lung cancer tissues (0.872±0.071) was significantly higher than that in paracancerous tissues (0.717±0.073) and benign pulmonary lesion tissues (0.611±0.026) (P < 0.001).(2)The transcription expression of endostatin mRNA in lung cancer tissues was closely related to P-TNM stages, distant metastasis, grade of cell differentiation and size of the primary tumors (P < 0.05), but not to location of tumor, lymph node status, histological classification, age and sex of the patients and smoking or not (P > 0.05).

CONCLUSIONSThe transcription expression of endostatin mRNA in NSCLC tissues is significantly higher than that in paracancerous tissues and benign pulmonary lesion tissues, and is closely related to P-TNM stages, distant metastasis, grade of cell differentiation and size of the primary tumors, hence it might be helpful to evaluate the biological behavior of lung cancer.