Astrocyte-Neuronal Communication and Its Role in Stroke.

Astrocytes are the most abundant glial cells in the central nervous system. These cells are an important hub for intercellular communication. They participate in various pathophysiological processes, including synaptogenesis, metabolic transformation, scar production, and blood-brain barrier repair. The mechanisms and functional consequences of astrocyte-neuron signaling are more complex than previously thought. Stroke is a disease associated with neurons in which astrocytes also play an important role. Astrocytes respond to the alterations in the brain microenvironment after stroke, providing required substances to neurons. However, they can also have harmful effects. In this review, we have summarized the function of astrocytes, their association with neurons, and two paradigms of the inflammatory response, which suggest that targeting astrocytes may be an effective strategy for treating stroke.

Chemical constituents from the twigs and leaves of Picrasma quassioides.

Two new compounds, including a norsesquiterpenoid, annuionone H (1), and a quassinoid, picraqualide G (2), along with eleven known compounds (3-13), were isolated from the twigs and leaves of Picrasma quassioides. Comprehensive spectroscopic analyses and NMR calculation with DP4+ analysis were used to identify their structures. Moreover, of all these compounds, compound 4 showed a week inhibition rate in the anti-inflammatory screening results against mouse macrophage J774A.1 cell.

Procyanidins Alleviated Cerebral Ischemia/Reperfusion Injury by Inhibiting Ferroptosis via the Nrf2/HO-1 Signaling Pathway.

Procyanidins (PCs), which are organic antioxidants, suppress oxidative stress, exhibit anti-apoptotic properties, and chelate metal ions. The potential defense mechanism of PCs against cerebral ischemia/reperfusion injury (CIRI) was investigated in this study. Pre-administration for 7 days of a PC enhanced nerve function and decreased cerebellar infarct volume in a mouse middle cerebral artery embolization paradigm. In addition, mitochondrial ferroptosis was enhanced, exhibited by mitochondrial shrinkage and roundness, increased membrane density, and reduced or absent ridges. The level of Fe2+ and lipid peroxidation that cause ferroptosis was significantly reduced by PC administration. According to the Western blot findings, PCs altered the expression of proteins associated with ferroptosis, promoting the expression of GPX4 and SLC7A11 while reducing the expression of TFR1, hence inhibiting ferroptosis. Moreover, the treatment of PCs markedly elevated the expression of HO-1 and Nuclear-Nrf2. The PCs' ability to prevent ferroptosis due to CIRI was decreased by the Nrf2 inhibitor ML385. Our findings showed that the protective effect of PCs may be achieved via activation of the Nrf2/HO-1 pathway and inhibiting ferroptosis. This study provides a new perspective on the treatment of CIRI with PCs.

Risk stratification in GIST: shape quantification with CT is a predictive factor.

BACKGROUND: Tumor shape is strongly associated with some tumor's genomic subtypes and patient outcomes. Our purpose is to find the relationship between risk stratification and the shape of GISTs. METHODS: A total of 101 patients with primary GISTs were confirmed by pathology and immunohistochemistry and underwent enhanced CT examination. All lesions' pathologic sizes were 1 to 10 cm. Points A and B were the extremities of the longest diameter (LD) of the tumor and points C and D the extremities of the small axis, which was the longest diameter perpendicular to AB. The four angles of the quadrangle ABCD were measured and each angle named by its summit (A, B, C, D). For regular lesions, we took angles A and B as big angle (BiA) and small angle (SmA). For irregular lesions, we compared A/B ratio and D/C ratio and selected the larger ratio for analysis. The chi-square test, t test, ROC analysis, and hierarchical or binary logistic regression analysis were used to analyze the data. RESULTS: The BiA/SmA ratio was an independent predictor for risk level of GISTs (p = 0.019). With threshold of BiA at 90.5°, BiA/SmA ratio at 1.35 and LD at 6.15 cm, the sensitivities for high-risk GISTs were 82.4%, 85.3%, and 83.8%, respectively; the specificities were 87.1%, 71%, and 77.4%, respectively; and the AUCs were 0.852, 0.818, and 0.844, respectively. LD could not effectively distinguish between intermediate-risk and high-risk GISTs, but BiA could (p < 0.05). Shape and Ki-67 were independent predictors of the mitotic value (p = 0.036 and p < 0.001, respectively), and the accuracy was 87.8%. CONCLUSIONS: Quantifying tumor shape has better predictive efficacy than LD in predicting the risk level and mitotic value of GISTs, especially for high-risk grading and mitotic value > 5/50HPF. KEY POINTS: • The BiA/SmA ratio was an independent predictor affecting the risk level of GISTs. LD could not effectively distinguish between intermediate-risk and high-risk GISTs, but BiA could. • Shape and Ki-67 were independent predictors of the mitotic value. • The method for quantifying the tumor shape has better predictive efficacy than LD in predicting the risk level and mitotic value of GISTs.

Effect of phase coarsening under melt annealing on the electrical performance of polymer composites with a double percolation structure.

The effect of phase coarsening on the evolution of the carbon black (CB) nanoparticle network under quiescent melt annealing and the electrical performance of polypropylene/polystyrene/carbon black (PP/PS/CB) composites with a double percolation structure was investigated. The results showed that when the CB content is low, the coarsening process of PP/PS/CB blends can be divided into two stages. In the first stage, the coarsening rate is fast before the formation of the CB nanoparticle network, and after annealing for a certain time, the evolution of the co-continuous morphology can drive the CB nanoparticles to self-assemble into a complete nanoparticle network. In the second stage, the coarsening rate is slow after the formation of the CB nanoparticle network. When the CB content is high, the CB nanoparticle network can be maintained throughout the whole annealing process, so that the conductivity and morphology of the PP/PS/CB composites are stable. Moreover, the electrical conductivity of the PP/PS/CB composites greatly increases after annealing for a certain time, and a percolation threshold as low as 0.07 vol% can be obtained. These results reveal the relationship between the evolution of the morphology and the conductivity in the conductive polymer composites with a double percolation structure, and provide a more in-depth and comprehensive understanding of the double percolation structure.

Decreased expression and DNA hypermethylation of syncytin-1 in human villus tissues with unexplained recurrent spontaneous abortion.

Recurrent spontaneous abortion (RSA) affects approximately 5 % of women of reproductive age worldwide. The etiology and pathogenesis of approximately 50 % of RSA cases currently remain unclear, which known as unexplained RSA (URSA). Syncytin-1, an envelope protein encoded by HERV-W gene, is essential for human embryonic development. The purpose of this study was to explore the correlation between syncytin-1 expression and URSA occurrence. The villi tissues of URSA patients and patients with voluntary termination of pregnancy for non-medical reasons in early pregnancy (Control group) were collected. Compared with the Control group, syncytin-1 was abnormally low expressed in URSA villus tissues, and the HERV-W gene promoter was hypermethylated. Compared with the control group, the global DNA methylation level and the expression level of DNA methylases in the villus tissues of the URSA group had no significant difference. In addition, compared with the Control group, URSA villus tissue showed obviously abnormal apoptosis. Overexpression of syncytin-1 promoted the proliferation of HTR-8 cells and inhibited their apoptosis; while knockdown of syncytin-1 inhibited cell proliferation and promoted cell apoptosis. URSA villus tissue exhibited hypermethylation of the HERV-W gene and down-regulation of syncytin-1 expression. Syncytin-1 has the potential to be a predictive and diagnostic biomarker for URSA.

Metabolic reprogramming regulates microglial polarization and its role in cerebral ischemia reperfusion.

The brain is quite sensitive to changes in energy supply because of its high energetic demand. Even small changes in energy metabolism may be the basis of impaired brain function, leading to the occurrence and development of cerebral ischemia/reperfusion (I/R) injury. Abundant evidence supports that metabolic defects of brain energy during the post-reperfusion period, especially low glucose oxidative metabolism and elevated glycolysis levels, which play a crucial role in cerebral I/R pathophysiology. Whereas research on brain energy metabolism dysfunction under the background of cerebral I/R mainly focuses on neurons, the research on the complexity of microglia energy metabolism in cerebral I/R is just emerging. As resident immune cells of the central nervous system, microglia activate rapidly and then transform into an M1 or M2 phenotype to correspond to changes in brain homeostasis during cerebral I/R injury. M1 microglia release proinflammatory factors to promote neuroinflammation, while M2 microglia play a neuroprotective role by secreting anti-inflammatory factors. The abnormal brain microenvironment promotes the metabolic reprogramming of microglia, which further affects the polarization state of microglia and disrupts the dynamic equilibrium of M1/M2, resulting in the aggravation of cerebral I/R injury. Increasing evidence suggests that metabolic reprogramming is a key driver of microglial inflammation. For example, M1 microglia preferentially produce energy through glycolysis, while M2 microglia provide energy primarily through oxidative phosphorylation. In this review, we highlight the emerging significance of regulating microglial energy metabolism in cerebral I/R injury.

The role of mitochondrial dynamics in cerebral ischemia-reperfusion injury.

Stroke is one of the leading causes of death and long-term disability worldwide. More than 80 % of strokes are ischemic, caused by an occlusion of cerebral arteries. Without question, restoration of blood supply as soon as possible is the first therapeutic strategy. Nonetheless paradoxically, reperfusion can further aggravate the injury through a series of reactions known as cerebral ischemia-reperfusion injury (CIRI). Mitochondria play a vital role in promoting nerve survival and neurological function recovery and mitochondrial dysfunction is considered one of the characteristics of CIRI. Neurons often die due to oxidative stress and an imbalance in energy metabolism following CIRI, and there is a strong association with mitochondrial dysfunction. Altered mitochondrial dynamics is the first reaction of mitochondrial stress. Mitochondrial dynamics refers to the maintenance of the integrity, distribution, and size of mitochondria as well as their ability to resist external stimuli through a continuous cycle of mitochondrial fission and fusion. Therefore, improving mitochondrial dynamics is a vital means of treating CIRI. This review discusses the relationship between mitochondria and CIRI and emphasizes improving mitochondrial dynamics as a potential therapeutic approach to improve the prognosis of CIRI.

Upregulation of CDGSH iron sulfur domain 2 attenuates cerebral ischemia/reperfusion injury.

CDGSH iron sulfur domain 2 can inhibit ferroptosis, which has been associated with cerebral ischemia/reperfusion, in individuals with head and neck cancer. Therefore, CDGSH iron sulfur domain 2 may be implicated in cerebral ischemia/reperfusion injury. To validate this hypothesis in the present study, we established mouse models of occlusion of the middle cerebral artery and HT22 cell models of oxygen-glucose deprivation and reoxygenation to mimic cerebral ischemia/reperfusion injury in vivo and in vitro, respectively. We found remarkably decreased CDGSH iron sulfur domain 2 expression in the mouse brain tissue and HT22 cells. When we used adeno-associated virus and plasmid to up-regulate CDGSH iron sulfur domain 2 expression in the brain tissue and HT22 cell models separately, mouse neurological dysfunction was greatly improved; the cerebral infarct volume was reduced; the survival rate of HT22 cells was increased; HT22 cell injury was alleviated; the expression of ferroptosis-related glutathione peroxidase 4, cystine-glutamate antiporter, and glutathione was increased; the levels of malondialdehyde, iron ions, and the expression of transferrin receptor 1 were decreased; and the expression of nuclear-factor E2-related factor 2/heme oxygenase 1 was increased. Inhibition of CDGSH iron sulfur domain 2 upregulation via the nuclear-factor E2-related factor 2 inhibitor ML385 in oxygen-glucose deprived and reoxygenated HT22 cells blocked the neuroprotective effects of CDGSH iron sulfur domain 2 up-regulation and the activation of the nuclear-factor E2-related factor 2/heme oxygenase 1 pathway. Our data indicate that the up-regulation of CDGSH iron sulfur domain 2 can attenuate cerebral ischemia/reperfusion injury, thus providing theoretical support from the perspectives of cytology and experimental zoology for the use of this protein as a therapeutic target in patients with cerebral ischemia/reperfusion injury.

16,17-dinor-abietane diterpenoids from Casearia kurzii.

Eleven undescribed 16,17-dinor-abietane diterpenoids, caseazins A-K (1-11), and ten known diterpenoids (12-21) were isolated from the twigs and leaves of Casearia kurzii (Flacourtiaceae). Caseazins A-K were the first abietane -type dinorditerpenoids to have been isolated from the plant of Casearia kurzii. Their chemical structures were elucidated using a combination of 1D and 2D NMR spectroscopy and mass spectrometry. The absolute configurations of 5 and 10 were established by electronic circular dichroism calculations. Moreover, compounds 2, 3, 13, 14, and 18 exhibited anti-inflammatory activity with IC50 values of 0.17, 0.36, 6.55, 1.30, and 4.53 µM, respectively. IL-1ß and caspase-1 analyses suggested that compound 14 inhibited NLRP3 inflammasome activation and blocked macrophage pyroptosis.

Nudifloids A-N, structurally diverse 3,4-seco-labdane diterpenoids from Callicarpa nudiflora with inflammatory inhibitory activity.

Fourteen undescribed seco-type diterpenoids, named nudifloids A-N, together with ten known analogs, were isolated from the leaves of Callicarpa nudiflora. Nudifloids A-N had a characteristic 3,4-seco-labdane-type diterpenoid skeleton, whereas nudifloids A-C and K-N were 3,4-seco-norditerpenoids. Nudifloid A was the first example of a 3,4-seco-12,13,14,15,16-quartnor-labdane diterpenoid, with a seven-membered lactone ring formed through esterification between C-3 and C-11. Nudifloids B and C were a pair of highly modified 3,4-seco-labdane nor-diterpenoid epimers, of which C-2 and C-18 were cyclized together to form a cyclohexene fragment. The structures of these undescribed diterpenoids were established by spectroscopic analysis and reference data. The anti-inflammatory activity of diterpenoids in rich yield was evaluated by analyzing the inhibition of lipopolysaccharide plus nigericin-induced pyroptosis in J774A.1 cells. Nudifloids D and E exhibited prominent anti-NLRP3 inflammasome activity, with IC50 values of 1.80 and 1.59 µM, respectively. Cell permeability assays revealed that nudifloid D inhibited pyroptosis, which could ameliorate inflammation by blocking the activation of the NLRP3 inflammasome.

Quality improvement of nursing on patients with complex cerebral arteriovenous malformation undergoing hybrid surgery: a prospective single-center study.

BACKGROUND: We sought to explore an optimal clinical nursing mode following a hybrid surgery for cerebral arteriovenous malformation. METHODS: Patients with complex cerebral arteriovenous malformations seen in our neurosurgery department from January 2016 to December 2017 were prospectively enrolled. The hybrid surgery protocol included "angiographic diagnosis, surgical resection, and intraoperative angiographic evaluation" and "angiographic diagnosis and embolization, surgical resection, and intraoperative angiographic evaluation". The patients were randomly stratified into intensive care group and routine care group. After surgery, intensive or routine care was provided, and the prognosis of patients was evaluated, with a subsequent comparative analysis. RESULTS: A total of 109 cases were divided into the routine nursing group (n = 54 cases) and intensive nursing group (n = 55 cases). There were no significant differences between the two groups in baseline data before surgery. Postoperative lung infection in the intensive nursing group was significantly less frequent than those in the routine nursing group (5.5% vs. 18.5%, P=0.039) with pulmonary infection and lower extremity venous thrombosis (5.5% vs. 24.1%, P=0.006). The average hospital stay in the intensive nursing group was 14.4 ± 5.78 days, which was significantly lower than that in the routine nursing group (19.3 ± 6.38 days, P=0.013). At 3 months' follow-up after surgery, the Generic Quality of Life Inventory-74 (GQOLI-74) dimension score and GQOLI-74 total score in the enhanced group were significantly better than those in the routine nursing group (P=0.017 and 0.023, respectively). CONCLUSIONS: Intensive postoperative nursing can improve the safety of patients after hybrid surgery, reduce the postoperative complications and the average length of hospital stay, and improve the quality of life of patients.

Biobinder Nanocoating for Upgrading the Assembling Structures of High-Capacity Composite Electrodes with a Robust Polymeric Artificial Solid Electrolyte Interphase.

The success of next-generation lithium-ion batteries (LIBs) fundamentally depends on the rational design of not only the microstructure of an individual component but the component assembling structures on the electrode level. However, building advanced assembling structures for especially high-capacity electrodes is an urgent but a challenging task due to the lacking of in-depth understanding and effective strategies. Here, we propose a functional nanocoating biobinder using the well-known poly(lactic acid) to address the above need. It is found that the composite electrodes with this nanocoating biobinder are upgraded with uniform and robust assembling structures, including the electron-transportation network, ion-transportation network, and interfaces. Importantly, the nanocoating finally works as a new type of polymeric artificial cathode electrolyte interphase (poly-CEI) to protect the active particles. Therefore, a remarkable improvement in the electrochemical performance has been achieved for high-capacity electrodes (LiFePO4, lithium nickel cobalt manganite (NCM), and lithium nickel cobalt aluminum acid (NCA)). In particular, the LFP cathode can deliver a high discharge capacity of 74.6 mA h g-1 at 10C and a high capacity retention of 95.5% even after 850 cycles at 2C. For NCA and NCM cathodes, the cycling stability is dramatically improved due to the protection by the poly-CEI. In short, this study may reshape the essential roles of a binder in composite electrodes by highlighting its critical link to assembling structures.

[A test of screening to predict lung cancer among dust-exposed tin miners with sputum imaging cytometry].

OBJECTIVE: To evaluate efficacy of sputum imaging cytometry in early diagnosis on lung cancer among tin miners exposed to dust and analyze possible risk factors related to lung cancer among tin miners. METHODS: From a cohort of tin miners in Guangxi Province, a total of 345 male tin miners higher than 45 years old and with high exposure to crystalline silica dust were randomly selected as the objects. Imaging cytometry was used to implement the sputum analysis for the screening on lung cancer according to the experience diagnostic standard. All objects were then followed up to the end of 2006. Clinical diagnosis of lung cancer was used as the golden standard to evaluate the efficacy of screening. RESULTS: From 1998 to 2006, 11 new cases were diagnosed as clinical lung cancer. Except of age and exposure to occupational hazards, smoking status (P = 0.0384) and mean smoking dose (P = 0.0078) were significantly associated with lung cancer, and the adjusted odds ratio of high level to the low was 18.21 (2.15 approximately 154.39). The sensitivity, specificity and Youden's index of the sputum imaging cytometry for the experience diagnosis were 27.3%, 83.9% and 11.2% respectively. According to the ROC curve analysis, area under ROC Curve (AUC) of C2.5 (the percentage when the DNA index ranged from 1.25 to 2.50) was 0.647 (0.525 approximately 0.768), with the optimal operating point (OOP) of 1.70%. Sensitivity, specificity, agreement rate, positive predictive value, negative predictive value and Youden's index for predicting lung cancers in high-exposure tin miners were found to be 72.7%, 62.3%, 62.6%, 6.0%, 98.6% and 35.0% respectively. CONCLUSION: Smoking is confirmed as an important risk factor of lung cancer in tin miners. The diagnostic efficiency can be improved if the diagnostic point of C2.5 is adjusted to 1.70%.

Using 18F-fluorodeoxyglucose positron emission tomography/computed tomography to estimate the length of gross tumor and involvement of lymph nodes in esophagogastric junction carcinoma.

OBJECTIVE: To determine the optimal approach for estimating the length of gross tumor and involvement of the lymph nodes with 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) in esophagogastric junction carcinoma (EGJC). The result was verified with pathologic examination. MATERIALS AND METHODS: Twenty patients with diagnosed and untreated EGJC were enrolled. The length of the gross tumor was measured using different approaches with PET/CT: Standardized uptake value (SUV) 1.5-5.5 in intervals of 1.0 and 10%-50% of maximum SUV (SUVmax) on 18F-FDG PET/CT in intervals of 10%. The results were expressed as L1.0-L5.0, and L10%-L50%, respectively. The pathological length of gross tumor (Lpath) was calculated based on the shrinkage ratio of primary tumor. The measurable lymph nodes were measured on PET/CT preoperatively, labeled during operation, and examined for pathology. RESULTS: Lpath was 6.87 ± 2.25 cm, L30% and L2.5 were 6.61 ± 1.76 cm and 7.56 ± 1.89 cm, respectively. L30% was closer to Lpath than other % SUVmax, L2.5 was closer to Lpath than other absolute SUV thresholds. The diagnostic performance of 18F-FDG PET/CT for lymph nodes was best at the cutoff SUV of 2.7, providing sensitivity of 70% and a specificity of 83.7% for detecting lymph node metastases. CONCLUSIONS: The tumor length with 30% SUVmax as the threshold was closest to the actual pathological length of EGJC. The diagnostic efficiency of 18F-FDG PET/CT was best at the cutoff SUVmax of 2.7 for detecting lymph node metastases in EGJC.

Further characterization of the epidermal growth factor receptor ligand 11C-PD153035.

BACKGROUND: (11)C-4-N-(3-bromoanilino)-6,7-dimethoxyquinazoline ((11)C-PD153035) has been reported as a tracer for imaging human tumors that overexpress epidermal growth factor receptor (EGFR). However it is still unclear whether (11)C-PD153035 uptake correlates with EGFR expression levels. The objective of this study was to investigate the relationship between (11)C-PD153035 accumulation and EGFR expression levels. METHODS: Synthesis of (11)C-PD153035 was performed in the Tracerlab FXc system. Accumulation of (11)C-PD153035 by MDA-MB-468, A549 and MDA-MB-231 cells was measured in vitro. There were six tumor-bearing mice in each group. (11)C-PD153035 uptake in tumors was determined by positron emission tomography/computed tomography (PET/CT). Tumor/normal muscle tissue (T/NT) analysis in PET images was applied to quantify the PET data. Sixty minutes after PET/CT scanning, the nude mice were sacrificed and the tumors were excised. The (11)C-PD153035 accumulation in different tumors was determined by a gamma counter. RESULTS: Close correlation existed between the uptake and the level of EGFR expression both in vitro and ex vivo (r(2) = 0.72, P < 0.001; r(2) = 0.63, P = 0.003). When the static T/NT analysis method was applied to analyze the PET data, the observed correlation was again excellent (r(2) = 0.70, P = 0.001). CONCLUSIONS: The uptake of PET tracer (11)C-PD153035 closely correlates with the EGFR expression levels in tumor cells. (11)C-PD153035 has the potential to yield useful information for both cancer diagnosis and therapy.

[Correlation between FDG PET /CT and the expression of glutl and ki-67 antigen in esophageal cancer].

OBJECTIVE: To evaluate the correlation between standardized uptake valus (SUV) of 18F-fluorodeoxyglucose (18 F-FDG) of tumor at PET/CT examination and the expression of glucose transporter-1 (Glutl) and Ki-67 in esophageal cancer. METHODS: 56 patients with esophageal cancer were evaluated with 18 F-FDG PET/CT examination before operation. The expression of Glut1 and Ki-67 antigen in the tumor tissues was detected by immunohistochemistry after operation. RESULTS: (1) Positive rate of Glutl and Ki-67 expression in esophageal cancer tissues was 100% , respectively. There was a positive correlation between the expression of Glutl and Ki-67 and the clinical stages and differentiation of the tumor. The more the tumor and the clinical stages were advanced and the lower was the tumor differentiation, the more Glutl and Ki-67 were expressed. (2) There were abnormal radioactive high uptake regions on PET/CT imaging of esophagus in the 56 patients, which were confirmed by pathology as the primary carcinoma. The SUV was higher than 2. 5. There was a gradually increasing tendency in SUV along with the lowering of the tumor differentiation and the advance of clinical stages. (3)There was a correlation between the expression of Glutl, Ki-67 and the SUV, the more Glutl and Ki-67 were expressed, the higher the SUV of tumor 18F-FDG at PET/CT examination was in esophageal tumor tissues. CONCLUSION: There is a widespread expression of Glutl in esophageal cancer tissues, and the SUV may be used to indirectly evaluate the proliferative capacity of esophageal cancer.

[Assessment of cervical intraepithelial neoplasia (CIN) lesions by DNA image cytometry].

OBJECTIVE: To compare the value of conventional cytology and DNA image cytometry (DNA-ICM) assisted cytology in detection and prognostic assessment of cervical CIN lesions. METHODS: 87 women were enrolled in this study. Cervical samples were collected employing cervix brushes which were then washed in Sedfix. After preparing single cell suspensions by mechanical procedure, cell monolayers were prepared by cyto-spinning the cells onto microscope slides. Two slides were prepared from each case: one slide was stained by Papanicolou staining for conventional cytology, another was stained by Feulgen-Thionin method for measurements of the amount of DNA in the cell nuclei using an automated DNA imaging cytometer. Biopsies from the cervical lesions were also taken for histopathology and Ki-67 immunohistochemistry. RESULTS: Of the total of 20 ASCUS cases called by conventional cytology, no CIN, nor greater lesions were found. Among the 20 cases, 7 cases did not show any cells with DNA amount greater than 5c, while CIN2 lesions were found in 11 of other 13 cases that had some aneuploid cells with DNA amount greater than 5c. Of 30 LSIL cases called by conventional cytology, CIN2 lesions were detected in 3 out of 7 cases that did not contain any aneuploid cells with DNA greater than 5c, but in 22 out of the other 23 cases that contained aneuploid cells with DNA amount greater than > 5c. Of the remaining 7 cases called HSIL by conventional cytology, all case contained aneuploid cells containing DNA greater than 5c. If cytology was used to refer all cases of LSIL and HSIL to colposcopy procedure to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive value were 58.2%, 84.4%, 86.5% and 54.0%, respectively. If DNA-ICM were used and all cases having 3 or more cells with a DNA amount greater than 5c were assessed to be referred to pathology to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive were 72.7% , 87.5%, 90.9% and 65.1%, respectively. We also compared Ki67 positive cells in these samples and found that DNA-ICM results were comparable to this biomarker method. CONCLUSION: The study demonstrated that DNA-ICM approach can be successfully used to detect significant (i.e. CIN2 or greater) lesions, and also provide a prognostic assessment of CIN lesions.

Detection of cervical cancer and high grade neoplastic lesions by a combination of liquid-based sampling preparation and DNA measurements using automated image cytometry.

OBJECTIVE: To establish if measurements of DNA ploidy could be used to assist cytopathologists and cytotechnologists in population based cervical cancer screening programs in countries where manually reading the slides is impossible due to the lack of sufficient skilled cytotechnologists. The goal of such program is to identify only clinically significant lesions, i.e. those where a clinical intervention to remove the lesion is required immediately. STUDY DESIGN: A total of 9905 women were enrolled in the study. Cervical samples were taken with a cervix brush that was then placed into a fixative solution. The cells were separated from mucus by mechanical and chemical treatment and then deposited onto microscope slides by a cytocentrifuge. Two slides were prepared from each case; one slide was stained by Papanicolaou stain for manual cytology examination, while the other slide was stained by a DNA specific stain. The latter slide was used to determine the relative amount of DNA in the cell nuclei. RESULTS: A total of 876 women were followed by colposcopy examination where biopsies were taken from the visible lesions or from suspicious areas and histopathology diagnosed 459 as normal or benign cases, 325 as CIN1, 36 as CIN2, 25 as CIN3/CIS, and 31 as invasive cancer. Of these 876 cases, manual cytology called 655 normal or ASCUS, 197 as LSIL, 16 cases as HSIL, and 8 as cancer. DNA measurements found 704 cases having no cells with DNA greater than 5c, 98 cases where there were 1 or 2 cells having DNA amount greater than 5c, and 74 cases where there were 3 or more cells having DNA amount greater than 5c. If manual cytology were to be used to refer all cases of HSIL and cancer to colposcopy and biopsy, 23 lesions that had to be removed would have been discovered (2 CIN2, 11 CIN3/CIS, and 10 cancers), for a sensitivity of 25.0+/-5.2% at specificity of 99.9+/-0.1%. If DNA assisted cytology were to be used instead, and all cases having 3 or more cells with DNA amount greater than 5c were to be referred to colposcopy and biopsy, then 50 lesions that had to be removed would have been discovered (10 CIN2, 15 CIN3/CIS and 25 cancers) for the sensitivity of 54.3+/-6.2% at specificity of 96.9+/-0.6%. CONCLUSIONS: The study suggests that screening for high grade cervical neoplastic lesions and cervical cancer by DNA assisted cytology could be implemented with minimal use of skilled cytotechnologists, at least in those countries where it would be difficult to introduce population based screening for cervical cancer due to the lack of availability of such skilled cytotechnologists.

Comparison of primary target volumes delineated on four-dimensional CT and 18 F-FDG PET/CT of non-small-cell lung cancer.

BACKGROUND: To determine the optimal threshold of 18 F-fluorodexyglucose (18 F-FDG) positron emission tomography CT (PET/CT) images that generates the best volumetric match to internal gross target volume (IGTV) based on four-dimensional CT (4DCT) images. METHODS: Twenty patients with non-small cell lung cancer (NSCLC) underwent enhanced three-dimensional CT (3DCT) scan followed by enhanced 4DCT scan of the thorax under normal free breathing with the administration of intravenous contrast agents. A total of 100 ml of ioversol was injected intravenously, 2 ml/s for 3DCT and 1 ml/s for 4DCT. Then 18 F-FDG PET/CT scan was performed based on the same positioning parameters (the same immobilization devices and identical position verified by laser localizer as well as skin marks). Gross target volumes (GTVs) of the primary tumor were contoured on the ten phases images of 4DCT to generate IGTV10. GTVPET were determined with eight different threshold using an auto-contouring function. The differences in the position, volume, concordance index (CI) and degree of inclusion (DI) of the targets between GTVPET and IGTV10 were compared. RESULTS: The images from seventeen patients were suitable for further analysis. Significant differences between the centric coordinate positions of GTVPET (excluding GTVPET15%) and IGTV10 were observed only in z axes (P < 0.05). GTVPET15%, GTVPET25% and GTVPET2.0 were not statistically different from IGTV10 (P < 0.05). GTVPET15% approximated closely to IGTV10 with median percentage volume changes of 4.86%. The best CI was between IGTV10 and GTVPET15% (0.57). The best DI of IGTV10 in GTVPET was IGTV10 in GTVPET15% (0.80). CONCLUSION: None of the PET-based contours had both close spatial and volumetric approximation to the 4DCT IGTV10. At present 3D-PET/CT should not be used for IGTV generation.