Development and Validation of a 18F-FDG PET-Based Radiomic Model for Evaluating Hypermetabolic Mediastinal-Hilar Lymph Nodes in Non-Small-Cell Lung Cancer.

BACKGROUND: Accurate evaluation of lymph node (LN) status is critical for determining the treatment options in patients with non-small cell lung cancer (NSCLC). This study aimed to develop and validate a 18F-FDG PET-based radiomic model for the identification of metastatic LNs from the hypermetabolic mediastinal-hilar LNs in NSCLC. METHODS: We retrospectively reviewed 259 patients with hypermetabolic LNs who underwent pretreatment 18F-FDG PET/CT and were pathologically confirmed as NSCLC from two centers. Two hundred twenty-eight LNs were allocated to a training cohort (LN = 159) and an internal validation cohort (LN = 69) from one center (7:3 ratio), and 60 LNs were enrolled to an external validation cohort from the other. Radiomic features were extracted from LNs of PET images. A PET radiomics signature was constructed by multivariable logistic regression after using the least absolute shrinkage and selection operator (LASSO) method with 10-fold cross-validation. The PET radiomics signature (model 1) and independent predictors from CT image features and clinical data (model 2) were incorporated into a combined model (model 3). A nomogram was plotted for the complex model, and the performance of the nomogram was assessed by its discrimination, calibration, and clinical usefulness. RESULTS: The area under the curve (AUC) values of model 1 were 0.820, 0.785, and 0.808 in the training, internal, and external validation cohorts, respectively, showing good diagnostic efficacy for lymph node metastasis (LNM). Furthermore, model 2 was able to discriminate metastatic LNs in the training (AUC 0.780), internal (AUC 0.794), and external validation cohorts (AUC 0.802), respectively. Model 3 showed optimal diagnostic performance among the three cohorts, with an AUC of 0.874, 0.845, and 0.841, respectively. The nomogram based on the model 3 showed good discrimination and calibration. CONCLUSIONS: Our study revealed that PET radiomics signature, especially when integrated with CT imaging features, showed the ability to identify true and false positives of mediastinal-hilar LNM detected by PET/CT in patients with NSCLC, which would help clinicians to make individual treatment decisions.

Upregulated expression of serum exosomal miR-375 and miR-1307 enhance the diagnostic power of CA125 for ovarian cancer.

BACKGROUND: Ovarian cancer (OC) is associated with high mortality in gynecological oncology; this is mainly due to the low diagnosis rate. Exosomal miRNA has demonstrated potential as a tumor biomarker. We aimed to explore the diagnostic potential of serum exosomal miR-1307 and miR-375 for OC. METHODS: The first six candidate miRNAs were selected from the previous literature. The relative quantification of qRT-PCR was used to screen for the stability of exosomal miRNAs, followed by validation of the cohort. ROC analysis was employed to analyze the specificity and sensitivity of exosomal miRNA. RESULTS: MiR-1307 and miR-375 were confirmed stably existing in serum exosomes of OC. Moreover, miR-1307 and miR-375 were both significantly up-regulated in serum exosomes of OC compared to ovarian benign and healthy groups. The overexpressed miRNAs showed independent diagnostic power and enhanced the diagnostic accuracy of traditional biomarkers when combined with CA-125 and HE4. MiR-1307 was associated with tumor staging, and miR-375 was associated with lymph node metastasis of OC. CONCLUSION: Our results suggest that serum exosomal miR-1307 and miR-375 could serve as potential tumor biomarkers to improve diagnostic efficiency for OC.

Outbreaks of acute gastroenteritis associated with a re-emerging GII.P16-GII.2 norovirus in the spring of 2017 in Jiangsu, China.

A total of 64 acute gastroenteritis outbreaks with 2,953 patients starting in December of 2016 and occurring mostly in the late spring of 2017 were reported in Jiangsu, China. A recombinant GII.P16-GII.2 norovirus variant was associated with 47 outbreaks (73.4%) for the gastroenteritis epidemic, predominantly occurring in February and March of 2017. Sequence analysis of the RNA-dependent RNA polymerase (RdRp) and capsid protein of the viral isolates from these outbreaks confirmed that this GII.P16-GII.2 strain was the GII.P16-GII.2 variant with the intergenotypic recombination, identified in Taiwan, Hong Kong, and other cities in China in 2016. This GII.P16-GII.2 recombinant variant appeared to a re-emerging strain, firstly identified in 2011-2012 from Japan and USA but might be independently originated from other GII.P16-GII.2 variants for sporadic and outbreaks of gastroenteritis in Japan and China before 2016. Further identification of unique amino acid mutations in both VP1 and RdRp of NoV strain as shown in this report may provide insight in explaining its structural and antigenic changes, potentially critical for the variant recombinant to gain its predominance in causing regional and worldwide epidemics.

Molecular epidemiology of genogroup II norovirus infection among hospitalized children with acute gastroenteritis in Suzhou (Jiangsu, China) from 2010 to 2013.

Noroviruses (NoVs) are the most common cause of acute gastroenteritis in both sporadic and outbreak cases. Genotyping and recombination analyses were performed in order to help getting more knowledge of the distribution and genetic diversity of NoVs in Suzhou, located in Jiangsu province of China. All stool samples were collected from hospitalized children younger than 5 years old with acute gastroenteritis. For genotyping, the open reading frame (ORF) 1 and ORF2 were partially amplified and sequenced. 26.9% of stool samples were positive for genogroup II NoVs. The most common genotype was GII.4 and its variants included Den Haag-2006b, New Orleans-2009, and Sydney-2012. The Den Haag-2006b variants predominated during 2010-2012. In 2013, it was replaced by the Sydney-2012 variant. The second most common genotype was GII.12/GII.3. NoVs could be detected throughout the year, with GII.4 and GII.12/GII.3 coexisting during the cold months, and GII.4 was the main genotype during the warm months. The highest prevalence of NoV was detected in young children aged <24 months. Patients infected with GII.4 had a higher chance of getting moderate fever than other NoV-positive patients, while those infected with GII.12/GII.3 tended to have a mild degree of fever. NoV is an important pathogen responsible for viral gastroenteritis among children in Suzhou. Analyses of NoV circulating between 2010 and 2013 revealed a change of predominant variant of NoV GII.4 in each epidemic season and intergenotype recombinant strains represented an important part.

Seroepidemiology of hepatitis B virus infection and impact of vaccination.

AIM: To investigate hepatitis B virus (HBV) prevalence in the general population in China. METHODS: A total of 148931 individuals were investigated by multistage random sampling in Eastern China. Data were collected on demographics and hepatitis B vaccination history, and serum was tested for hepatitis B surface antigen (HBsAg) by ELISA. RESULTS: A total of 11469 participants (7.70%, 95%CI: 7.57%-7.84%) were positive for HBsAg. HBsAg prevalence was 0.77% among children < 5 years old but increased progressively from adolescents (1.40%-2.55%) to adults (5.69%-11.22%). A decrease in HBsAg prevalence was strongly associated with vaccination and familial history of HBV among both children and adult groups. Meanwhile, HBsAg risk in adults was associated with invasive testing and sharing needles. The HBV immunization rate among participants aged < 20 years was 93.30% (95%CI: 93.01%-93.58%). Significant difference in HBsAg prevalence appeared between vaccinated and unvaccinated participants (3.59% vs 10.22%). CONCLUSION: Although the national goal of HBsAg prevalence < 1% among children < 5 years old has been reached, immunization programs should be maintained to prevent resurgence.

Epidemiological and clinical characteristics and risk factors for death of patients with avian influenza A H7N9 virus infection from Jiangsu Province, Eastern China.

BACKGROUND: A novel avian influenza A (H7N9) virus has caused great morbidity as well as mortality since its emergence in Eastern China in February 2013. However, the possible risk factors for death are not yet fully known. METHODS AND FINDINGS: Patients with H7N9 virus infection between March 1 and August 14, 2013 in Jiangsu province were enrolled. Data were collected with a standard form. Mean or percentage was used to describe the features, and Fisher's exact test or t-test test was used to compare the differences between fatal and nonfatal cases with H7N9 virus infection. A total of 28 patients with H7N9 virus infection were identified among whom, nine (32.1%) died. The median age of fatal cases was significant higher than nonfatal cases (P<0.05). Patients with older age were more strongly associated with increased odds of death (OR = 30.0; 95% CI, 2.85-315.62). Co-morbidity with chronic lung disease and hypertension were risk factors for mortality (OR = 14.40; 95% CI, 1.30-159.52, OR = 6.67; 95% CI, 1.09-40.43, respectively). Moreover, the presence of either bilateral lung inflammation or pulmonary consolidation on chest imaging on admission was related with fatal outcome (OR = 7.00; 95%CI, 1.10-44.61). Finally, dynamic monitoring showed that lymphopenia was more significant in fatal group than in nonfatal group from day 11 to week five (P<0.05). The decrease in oxygenation indexes were observed in most cases and more significantly in fatal cases after week three (P<0.05), and the value of nearly all fatal cases were below 200 mmHg during our evaluation period. CONCLUSIONS: Among cases with H7N9 virus infection, increased age accompanied by co-morbidities was the risk of death. The severity of lung infection at admission, the persistence of lymphocytopenia, and the extended duration of lower oxygenation index all contributed to worsened outcomes of patients with H7N9 virus infection.

BF3·Et2O-catalyzed formal [3 + 2] reaction of aziridinofullerenes with carbonyl compounds.

The BF3·Et2O-catalyzed formal [3 + 2] reaction of aziridinofullerenes with various carbonyl compounds for the easy preparation of fullerooxazolidines has been developed. Moreover, the reaction of aziridinofullerene with ethyl formate affords the simplest fullerooxazole without substituent.

Emergence of two novel norovirus genotype II.4 variants associated with viral gastroenteritis in China.

Noroviruses (NoVs) are the principal cause of epidemic viral gastroenteritis worldwide, including industrialized and developing countries. Eight hundred and fifty sporadic specimens from hospitalized children with acute gastroenteritis and 131 specimens from seven gastroenteritis outbreaks were collected during 2011-2012 in Jiangsu, China. All specimens were tested for the presence of norovirus (NoV) by real time RT-PCR, and in these, 225/850 of sporadic specimens and 76/131 of outbreak specimens were positive. By sequencing, two novel variants termed JS2011/CHN variant and JS2012/CHN variant were found. By complete genome sequencing and phylogenetic analysis confirmed that both JS2011/CHN variant and JS2012/CHN variant shared more than 98% identity with GII.4 New Orleans/2009/USA strain and GII.4 Sydney/2012/AUS. Both of them had mutations in some key sites in nucleotide sequence and amino acid sequence of ORF1-ORF3. Whether two novel variants will cause epidemic of NoV outbreaks in China deserves further attention. A national surveillance network may be needed to identify trends in molecular evolution of NoVs for prevention of future epidemics.

Spatial analyses of typhoid fever in Jiangsu province, People's Republic of China.

An analysis of the geographical distribution of typhoid incidence rates, based on various statistical approaches such as trend surface, spatial autocorrelation, spatial correlation and spatial regression, was carried out at the county level in Jiangsu province, People's Republic of China. Temperature, moisture content, proximity to water bodies and the normalized difference vegetation index in the autumn were the four underlying factors found to contribute the most to the development of the epidemic. Typhoid infection was most severe in the south-eastern region of Jiangsu and a significant hotspot with high positive autocorrelation was detected in Taicang county in the south-east of the province. To improve the typhoid situation, intervention efforts should be concentrated in the south-eastern region of the province, targeting the hotspot and include reduction of lake pollution.

[Molecular features of human bocavirus from infantile diarrhea in Suzhou area, Jiangsu province in 2010-2011].

OBJECTIVE: To investigate the status of human bocavirus and to identify its epidemiological characteristics as well as genotype distribution in patients with infantile viral diarrhea in Suzhou, Jiangsu province. METHODS: 832 fecal specimens from patients with infantile virus diarrhea cases were collected from Suzhou Children's Hospital in 2010-2011. Human bocavirus were detected by Real-Time RT-PCR, and genotype were determined by sequence analysis. RESULTS: Among all the fecal specimens, 51 (6.1%) cases were positive for human bocavirus. The peak season of rotavirus infection was between July and September. Of all the episodes on rotavirus diarrhea, 96% occurred before 2 years of age, with peaks in children with 7-12 months of age. Data from Nucleotide Sequence analysis showed that among 28 samples that carrying HBoV-1, 5 strains belonged to HBoV-2, HBoV type 3 but type 4 were absent. CONCLUSION: Human bocavirus were detected from fecal specimens of infantile virus diarrhea in Suzhou, with genotype HBoV-1 as the major strain. HBoV-2 genotype was also found.

[Molecular characteristics of acute gastroenteritis outbreaks caused by norovirus, in Jiangsu province].

OBJECTIVE: To study both the epidemiologic and molecular characteristics of outbreaks caused by norovirus (NoV) with its variants, in Jiangsu. METHODS: 67 specimens from seven gastroenteritis outbreaks were collected from October 2012 to March 2013 in Jiangsu. NoV gene group was detected by Real-Time RT-PCR. NoV portions of RdRp gene and VP1 gene were amplified under RT-PCR. RESULTS: Seven gastroenteritis outbreaks were caused by NoV. Among all the fecal specimens,45 (67.2%) showed positive to NoV G II. Study on the genotype was conducted through analyzing the nucleotide sequence of RdRp gene. Based on the RdRp region, 7 strains appeared to be G II, with 3 and 38 strains belonged to G II.4--Sydney variants. Results from phylogenetic analysis confirmed that 38 variants shared 99% identity with G II.4--Sydney. We also amplified the VP1 genes from 6 variants and comparing with 9 epidemic strains on the sequence amino acid sequence. All the strains showed mutation in amino acid sequence at some key sites which were closely related to the forming of neutralizing epitopes. CONCLUSION: The short interval periods between all 7 NoV outbreaks with identical viral strain indicated the emergence of a new NoV variant in Jiangsu province,that had caused a number of epidemics abroad. Results from our study suggested that the development of monitoring programs on this novel G II.4--Sydney variant should be a part of the NoV surveillance in Jiangsu province or even in the country.

[Analysis on the epidemiological and genetic characteristics of enterovirus type 71 and Coxsackie A16 virus infection in Jiangsu, China].

OBJECTIVE: To determine the epidemiological features of hand-foot-and-mouth disease (HFMD) outbreaks and the genetic characteristics of enterovirus type 71(EV71) isolates from patients in Lianyungang, Jiangsu province in May, 2008. METHODS: Epidemiological, microbiological, cellular and molecular methods were performed to investigate pathogens and to describe the homogeneity of isolated strains. RESULTS: 21 cases were reported in this HFMD outbreak with the attack rate as 20.0%. 3 EV71 virus strains were isolated from 10 stool samples. The nucleotide and amino acid homogeneity of these 3 Jiangsu strain with Anhui Fuyang strains were 97.9%-100.0% and 99.7%-100.0%, respectively. These 4 Jiangsu strains were within genotype C sub-geno group C4 in phylogenetic tree. Data from the follow-up study showed that shedding of EV71 and Coxsackie A16 virus (CA16) in the latent period appeared in the outbreak of HFMD. Human beings could be infected by both EV71 virus and CA16 at the same time and could also carry the two viruses. We also discovered that EV71 virus could be expelled out of the human body through stool in the first week and last for 10 weeks. CONCLUSION: The recently identified EV71 isolates from this HFMD outbreak belonged to sub-geno group C4. Facts as: the release of viruses in the latent period, co-infection or coexisting of two viruses at the same time and super long period of expulsion of toxin exist in EV71 and CA16 did exist.

Proteomic analysis of effect of hyperthermia on spermatogenesis in adult male mice.

We characterized cellular and molecular mechanisms involved in spermatogenesis following short-term heat exposure of murine testis. For these studies, we utilized a proteomic approach with two-dimensional gel electrophoresis (2DE) analyses and mass spectroscopic identification of proteins with altered expression in mouse testes at different times after heat shock. We established a proteome reference map from 7-wk-old mouse testis linked to a federated proteome database. We used these tools to analyze quantitative variations in the tissue over a time course of 0.5, 2, 6, and 12 h following heat exposure. We separated 108 protein spots expressed differentially between the heat shock tissues and the control mouse testes. Of these spots, we identified 36 by comparing with the control reference map. We then focused on the heterogeneous nuclear ribonucleoproteins (hnRNPs) and the chaperonins containing t-complex polypeptide-1 (CCT). Further analysis in this heat-shocked model suggests numerous potential mechanisms for heat shock-induced spermatogenic disorder.

Fungal spectrum identified by a new slide culture and in vitro drug susceptibility using Etest in fungal keratitis.

PURPOSE: To investigate the spectrum of fungal species causing keratitis and to test antifungal drug susceptibility to each isolate using Etest. METHODS: Microbial cultures were performed for patients who were clinically diagnosed with fungal keratitis between September 2002 and July 2004. Modified slide culture was established to identify the fungal species of the isolates. Etest (AB BIODISK, Solna, Sweden) was applied to determine the antifungal agent susceptibility of each isolate to itraconazole, fluconazole, and amphotericin B in vitro, respectively. RESULTS: Among 73 eyes of 73 patients with clinical diagnosis of fungal keratitis, 61 strains of fungi were isolated from 61 eyes. The rate of positive culture was 81.3% of all cases. The spectrum of fungal species involved: 58 (95.1%) isolates of filamentous fungi, including the two most common genera-Fusarium (n = 33, 54.1%) and Aspergillus (n = 9, 14.8%),-followed by 16 (26.2%) isolates of other genera of filamentous fungi such as Alternaria (n = 3, 4.9%), Trichophyton (n = 3, 4.9%), Curvularia (n = 2, 3.3%), Chrysosporium (n = 2, 3.3%), Acremonium (n = 2, 3.3%), and Scedosporium (n = 1, 1.6%), 1 (1.6%) yeast of Candida, as well as two (3.3%) dimorphic fungi of Blastomyces and Sporothrix isolate each. Three filamentous fungi of the isolates failed to be identified according to the information provided by slide culture. The results of Etest showed that 20 (60.6%) isolates of Fusarium were susceptible to amphotericin B, whereas all of them were resistant to itraconazole and fluconazole. All nine (100%) isolates of Aspergillus were sensitive to itraconazole, whereas four (44.4%) of them were sensitive to amphotericin B, and only two (22.2%) of them were sensitive to fluconazole. Seventeen (89.5%), 13 (68.4%), and 10 (52.6%) isolates of the remaining 19 organisms were sensitive to amphotericin B, itraconazole, and fluconazole, respectively. CONCLUSIONS: Fusarium and Aspergillus are the most frequent pathogenic organisms in causing fungal keratitis, whereas other species of fungi can also cause corneal infection. In vitro Etest for assessing antifungal drug susceptibility is a simple and practical method and may provide referential information for clinical consideration of choosing antifungal agents to treat fungal keratitis.

A two-dimensional electrophoresis reference map of human ovary.

The ovary plays a central role in oogenesis and gonadal hormone secretion. Proteomic analysis is a valuable approach for gaining an increased understanding of the molecular nature of the ovary. In this work, two-dimensional electrophoresis for protein separation followed by matrix-assisted laser desorption/ionization mass spectrometry and database searches, identified 231 protein spots corresponding to 138 individual proteins that were found in gels representing both the follicular and luteal phases. The data were used to construct a database online (http://reprod.njmu.edu.cn/2d). The identified proteins were functionally classified into seven groups: (1) cell signaling/communication, (2) cell division, (3) gene/protein expression, (4) metabolism, (5) cell structure and motility, (6) cell/organism defense, and (7) unclassified. Among the proteins identified, 47% had not been previously reported in the human ovary. In addition, a number of disease-related proteins were identified in this protein map, including some cancer- and polycystic ovarian syndrome-related proteins. Two proteins with phosphorylation were verified by Western blot analysis. Comparison of protein abundance between follicular and luteal stages produced seven protein spots that had been identified in our database. This study provides a preliminary reference map of normal human ovary that will form a basis for comparative studies on normal and pathological conditions of the human ovary and may serve as a potential tool for clinical diagnosis, therapeutics, and prognosis.

[A two-dimensional reference map of mouse ovary proteins].

OBJECTIVE: To perform a preliminary proteomic analysis of mouse ovaries and to study the protein's function in mouse ovary. METHODS: The two-dimensional gel electrophoresis (2-DE) and matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) were used to analyze mouse ovarian proteome. A 12.5% sodium dodecyl sulfate (SDS) reference gel was generated by immobilized pH gradient isoelectric focusing of mouse ovary proteins in a non-linear gradient (pH 3-10). And GRP78 was selected to perform with immunohistochemistry within mouse ovaries. RESULTS: Based on peptide mass fingerprinting, 52 proteins were identified and classified into seven functional groups: Cell/organism defense and antioxidant, cell signaling/communications proteins, cell structure/motility proteins, metabolism proteins, RNA synthesis processing, protein synthesis and processing, and unclassified proteins. The immunoreactivity of GRP78 was detected in GCs in the follicular, and with during GCs Luteinizing in the menstrual cycle, the protein expression (brown) increased continually and came to a head when ovulation happened. CONCLUSION: This work provides a first step toward the establishment of a systematic ovary protein database and stands as a valuable resource for molecular analyses of normal and pathologic conditions affecting mouse ovaries.

Differential expression of glucose-regulated protein 78 during spermatogenesis.

The aim of the present study was to isolate and identify proteins involved in the process of spermatogenesis. To achieve this goal we used the technique of proteomic analysis. Comparison of testis protein patterns obtained by high-resolution two-dimensional gel electrophoresis from 1-week- and 7-week-old mice showed significant differences in protein spot intensities. Subsequently several of these variant protein spots were identified by mass spectrometry. Glucose-regulated protein (GRP) 78 (Bip) was one of them. GRP78, expressed at a lower level in 1-week-old mouse testes compared to 7-week-old mouse testes, is a member of the heat shock 70 protein family. It has recently been shown to be important for protecting cells from apoptosis in somatic cells, especially in progressively growing tumor cells. Further, immunohistochemical (IHC) analyses of mouse and human testes sections were performed to determine the cellular distribution of this protein. A strong GRP78 staining was seen beginning with pachytene spermatocytes. These findings suggested that GRP78 might perform an important function in the process of spermatogenesis.

[An improved quantitative method for evaluating neurological deficits in mice with focal cerebral ischemia].

The purpose of this study was to develop a quantitative and objective method for evaluating neurological deficits in mice with focal cerebral ischemia. After middle cerebral artery occlusion (MCAO), the neurological deficits were evaluated 24 h later. We measured the mean angles, dominant angles and turns in a hanged test in which the mice were sticked on the wall, and the holding angles in an inclined plane test as well, Then we determined the cerebral infarct volumes, neuron density in hippocampus, cortex and subcortical areas 24 h after MCAO. The correlations among infarct volume, neuron density and neurological deficits were analyzed. We also compared the quantitative method with two typical complex methods of behavioral assessment. The effect of [pranlukast, 4-oxo-8-[p-(4-phenylbutyloxy) benzoylamino]-2-(tetrazol-5-yl)-4H-1-benzopyran hemihydrate] (ONO-1078), a neuroprotective agent, on ischemic injury was observed using this method. We found that the variables measured by both quantitative and typical behavioral methods significantly changed in the ischemic mice, and correlated with the infarct volumes and neuron densities. The quantitative variables well correlated with those of typical behavioral assessment, too. ONO-1078 inhibited ischemic injury and reduced the total scores of quantitative assessment. Thus, the quantitative method we developed is useful in evaluating neurological deficits of focal cerebral ischemia with the advantages of objectivity, quantification, simplicity and non-invasion, and can be used in the evaluation of neuroprotective effects of drugs.

[Application of two-dimensional electrophoresis and mass spectrometry in the research of testicular proteins].

OBJECTIVES: To evaluate the application of two-dimensional electrophoresis and mass spectrometry in the research of proteins expressed in testis. METHODS: Protein from adult ICR mouse testes was extracted by two-dimensional electrophoresis. Two protein spots were cut from the gel, then the proteins were digested in-gel by enzyme and the generated peptides were measured by matrix assisted laser desorption/ionization-time of flight mass spectrometry. The proteins were identified through database searching. RESULTS: Two spots in Commasie Brilliant Blue-stained gel were identified as serum albumin and protein disulfide isomerase by database searching. CONCLUSIONS: This rapid high resolution and efficient method is a very powerful way to analyze testicular proteins.