A Highly Sensitive and Robust Method for Hepatitis B Virus Covalently Closed Circular DNA Detection in Single Cells and Serum.

Despite implications of persistence of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) in the development of hepatocellular carcinoma (HCC), little is known about serum cccDNA in HBV-infected diseases. We developed a cccDNA-selective droplet digital PCR (ddPCR) to assess cccDNA content and dynamics across different stages of HCC development. One hundred forty-seven serum samples and 35 formalin-fixed, paraffin-embedded tumor tissues were derived from patients with HCC or HBV hepatitis/cirrhosis. After specific amplification and selective digestion, probe-based ddPCR was used to quantify cccDNA copy numbers in single cells and clinical samples. The cccDNA in single HepG2.2.15 cells ranged from 0 to 10.8 copies/cell. Compared with non-HCC patients, HCC patients showed a higher cccDNA-positive rate (89.9% versus 53.2%; P = 4.22 × 10-6) and increased serum cccDNA contents (P = 0.002 and P = 0.041 for hepatitis and cirrhosis patients, respectively). Serum cccDNA ranged from 84 to 1.07 × 105 copies/mL. Quantification of serum cccDNA and HBV-DNA was an effective way to discriminate HCC patients from non-HCC patients, with areas under the curve of receiver operating characteristic of 0.847 (95% CI, 0.759-0.935; sensitivity, 74.5%; specificity, 93.7%). cccDNA-selective ddPCR is sensitive to detect cccDNA in single cells and different clinical samples. Combined analysis of serum cccDNA and HBV-DNA may be a promising strategy for HBV-induced HCC surveillance and antiviral therapy evaluation.

Extrapleural locating method in computed tomography-guided needle biopsies of 1,106 lung lesions.

Transthoracic needle biopsy is considered to be safe and effective for the diagnosis of focal lung lesions. The aim of the present study was to evaluate factors affecting the accuracy and safety of automated cutting needle lung biopsy (ACNB) using a new extrapleural locating (EPL) method. Computed tomography (CT)-guided needle biopsies were performed on 1,065 patients between March 2005 and May 2012 using the EPL method. The locating needle remained in the chest following extrapleural positioning, while the radiologist confirmed the puncture angle and distance between the locating needle and lesion. The biopsy instrument was advanced into the lung, and the core needle was subsequently fired into the lesion based on the direction indicated by the locating needle. Univariate and multivariate regression analyses were used to evaluate the diagnostic accuracy and safety of the procedure. The sensitivity, specificity, positive predictive value and negative predictive value of the extrapleural method were 91.9, 100, 100 and 82.9%, respectively, and the overall diagnostic accuracy was 94.2%. Significant risk factors affecting accuracy were younger age, atelectasis, hemoptysis and lesion depth (P<0.03). Multivariate logistic regression analysis revealed that the risk of malignant lesions receiving a false-negative diagnosis decreased for each additional year of subject age [odds ratio (OR), 0.97; P=0.027] and increased with each millimeter increase in lesion depth (OR, 1.03; P=0.008). Among the 1,106 lesions biopsied, 207 were associated with pneumothorax, 251 with hemorrhage and 58 with hemoptysis. Multivariate analysis revealed that lesion size and emphysema affected pneumothorax incidence, while age, lesion location and depth and emphysema significantly affected hemorrhage incidence (P<0.05). In conclusion, low-dose, CT-guided ACNB with the EPL method provides a safe and accurate diagnosis.

Clinical utility of dual-energy CT for gout diagnosis.

OBJECTIVE: The present study was to evaluate the clinical value of dual-energy computed tomography (DECT) for detecting monosodium urate crystals in patients with gouty arthritis. METHODS: Two hundred and two patients, who experienced arthrocele and (or) joint pain, were enrolled into our study. DECT scans of upper or lower extremity were performed. One hundred and sixty one patients who conformed to the American College of Rheumatology classification standard were defined as the gout group. The rest (41) of the patients were regarded as the without-gout group. DE (80kV/140kV) datasets were reconstructed via DE gout software. Images were reviewed independently by two senior radiologists. RESULTS: In the gout group, DECT scans revealed a total of 379 areas of urate deposition in 121 patients. In the without-gout group, 3 areas of green urate deposition were detected. The sensitivity and specificity were 75.2% and 92.7%, respectively; when we increased the ratio to 1.32 and decreased the range to 3, the number of patients with green urate deposition increased, and the areas of urate deposition were more extensive. The sensitivity and specificity were 91.9% and 85.4%. DECT images could illustrate the palpable reduction in the tissue urate deposits compared to baseline images before and after treatment. CONCLUSIONS: DECT has comparable sensitivity and specificity for the detection of gouty arthritis in a clinical setting, and DECT can monitor the clinical treatment. However, DECT results should be interpreted carefully because there could be some false-negative or false-positive findings.

Computed tomography-guided cutting needle pleural biopsy: Accuracy and complications.

In cases of pleural lesion, tissue samples can be obtained through thoracoscopy or closed needle biopsy for histopathological analysis. Cutting needle biopsy is a relatively recent addition to these techniques. The aim of this study was to evaluate the diagnostic accuracy and safety of computed tomography-guided cutting needle pleural biopsy (CT-CNPB), as well as the associated complications, in patients with pleural lesion. This study was a retrospective analysis of 92 percutaneous CT-CNPBs on 90 patients between March 2008 and May 2013. For group comparisons, χ2 tests were used to detect the risk factors for diagnostic accuracy (false-negative rate). Of the 92 CT-CNPBs, malignant lesions were diagnosed in 55 cases (mesothelioma in 12, metastatic pleural disease in 36, synoviosarcoma in one, indeterminate-origin disease in one and false-negative lesion in five) and benign pleural disease was diagnosed in 37 cases (inflammation in 15, tuberculosis in 10, granuloma in three, solitary fibrous tumor in two, hematoma in one, fungus in one and indeterminate-origin disease in five). The sensitivity of diagnostic malignant lesion was 90.9%, and the specificity and positive and negative predictive values were 100, 100 and 88.1%, respectively. The overall diagnostic accuracy was 94.6%. A specific diagnosis was achieved in 89.1% of malignant lesions and 86.4% of benign lesions. Univariate analysis of the risk factors affecting accuracy (false-negative rate) did not reveal any significant differences (all P>0.05). The complication rates were 6.5% for pneumothorax, 8.7% for hemorrhage and 1.1% for hemothorax. In conclusion, CT-CNPB is a safe and accurate diagnostic technique that can be recommended as the primary method of diagnosis in patients with pleural thickening or lesions observed by CT scan.

Primary hepatic peripheral T-cell lymphoma: A case report.

Primary hepatic peripheral T-cell lymphoma (PHL) is extremely rare. A case of primary hepatic peripheral T-cell lymphoma of a 59-year-old male is presented in the current study. PHL lesions are diagnosed by the existence of a hepatic mass, in the absence of lymphadenopathy, splenomegaly or bone marrow involvement associated with normal tumor markers. Treatment options are surgical resection and subsequent chemotherapy. Histopathological examination by immunohistochemical staining of the tissue biopsies at laparotomy confirmed a diagnosis of PHL.

Primary retroperitoneal extraskeletal mesenchymal chondrosarcoma involving the vena cava: A case report.

The current study presents a case of extraskeletal mesenchymal chondrosarcoma (ESMC) involving the vena cava that originally occurred in the retroperitoneum of a 61-year-old female. Following excision of the masses, pathological examination confirmed a diagnosis of primary ESMC. Mesenchymal chondrosarcomas are extremely rare in comparison to conventional chondrosarcomas and even more so when arising in an extraskeletal location. In the current report, the major characteristics of ESMC are discussed and a review of the current knowledge regarding this rare disease entity is presented.

Preoperative CT findings of thymoma are correlated with postoperative Masaoka clinical stage.

RATIONALE AND OBJECTIVES: Both preoperative computed tomography (CT) staging and postoperative surgical Masaoka clinical staging are of great clinical importance for diagnosing thymomas. Our study aimed to investigate the relationships between these two staging systems. MATERIALS AND METHODS: This was a retrospective review of 129 patients who had undergone thymoma surgery. Helical CT and 16-slice CT were performed preoperatively. Surgical findings were evaluated according to the Masaoka clinical staging system. RESULTS: A significant association was shown between Masaoka clinical staging and CT staging, especially of features including tumor size (P = .004), tumor shape (P < .001), tumor density (P < .001), capsule completeness (P < .001), and involvement of surrounding tissues (P < .001). Based on the CT findings, there were 35.09% of Masaoka stage I patients who had a tumor size <5 cm as compared to 14.81% of stage IV patients. Only 8.77% of Masaoka stage I patients had a tumor size ≥10 cm as compared to 40.74% of stage IV patients. In stages III and IV, most tumors were irregularly shaped with an uneven density and incomplete capsule. Invasive tumors were more frequently found in stages III (81.48%) and IV (88.89%) than in stages I (0%) and II (38.89%). The incidence of myasthenia gravis was comparable in different stages. Consistency between CT and Masaoka clinical stages was higher in stage I (37.98%) than other stages (approximately 10%). CONCLUSION: This study documented a close relationship between preoperative CT thymoma staging and postoperative Masaoka clinical staging. Thus, preoperative CT findings can be beneficial for determining the proper management and prognosis of thymoma patients.

Identification of cisplatin-resistance associated genes through proteomic analysis of human ovarian cancer cells and a cisplatin-resistant subline.

Chemoresistance to cancer therapy is a major obstacle to the effective treatment of human cancers with cisplatin (DDP), but the mechanisms of cisplatin-resistance are not clear. In this study, we established a cisplatin- resistant human ovarian cancer cell line (COC1/DDP) and identified differentially expressed proteins related to cisplatin resistance. The proteomic expression profiles in COC1 before and after DDP treatment were examined using 2-dimensional electrophoresis technology. Differentially expressed proteins were identified using matrix- assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and high performance liquid chromatography-electrospray tandem MS (NanoUPLC-ESI-MS/MS). 5 protein spots, for cytokeratin 9, keratin 1, deoxyuridine triphosphatase (dUTPase), aarF domain containing kinase 4 (ADCK 4) and cofilin1, were identified to be significantly changed in COC1/DDP compared with its parental cells. The expression of these five proteins was further validated by quantitative PCR and Western blotting, confirming the results of proteomic analysis. Further research on these proteins may help to identify novel resistant biomarkers or reveal the mechanism of cisplatin-resistance in human ovarian cancers.

Relationship between computed tomography manifestations of thymic epithelial tumors and the WHO pathological classification.

OBJECTIVE: To explore the relationship between computed tomography (CT) manifestations of thymoma and its WHO pathological classification. METHODS: One hundred and five histopathologically confirmed cases were collected for their pathological and CT characteristics and results were statistically compared between different pathological types of thymoma. RESULTS: Tumor size, shape, necrosis or cystic change, capsule integrity, invasion to the adjacent tissue, lymphadenopathy, and the presence of pleural effusion were significantly different between different pathological types of thymomas (P<0.05). Type B2, B3 tumors and thymic carcinomas were greater in size than other types. More than 50% of type B3 tumors and thymic carcinomas had a tumor size greater than 10 cm. The shape of types A, AB, and B1 tumors were mostly round or oval, whereas 75% of type B3 tumors and 85% of thymic carcinomas were irregular in shape. Necrosis or cystic change occurred in 67% of type B3 thymomas and 57% of thymic carcinomas, respectively. The respective figures for capsule destruction were 83% and 100%. Increases in the degree of malignancy were associated with increases in the incidence of surrounding tissue invasion: 33%, 75%, and 81% in type B2, type B3, and thymic carcinomas, respectively. Pleural effusion occurred in 48% of thymic carcinomas, while calcification was observed mostly in type B thymomas. CONCLUSIONS: Different pathological types of thymic epithelial tumors have different CT manifestations. Distinctive CT features of thymomas may reflect their pathological types.

[The factor analysis of the incidence of complication in CT-guided lung automated cutting needle biopsy with extrapleural locating method].

OBJECTIVE: To evaluate the complication rate and analyze its impact of multiple facts of CT-guided percutaneous lung automated cutting needle biopsies (ACNB) with extrapleural locating method (EPL). METHODS: Retrospective study where information was obtained from the department of medical imaging, the sign of complication after 480 cases CT-guided ACNB with EPL was observed and its relationship with multiple factors were analyzed by multiple logistic regression model. RESULTS: The diagnostic accuracy was made in 456 cases (95.0%). The length of the biopsy procedures was 16 ± 2 min and the time of the biopsy needle in pulmonary parenchyma was < 20 s. Seventy-one (14.8%) cases presented pneumothorax which include 7 (1.5%) cases later pneumothorax, and 2 (0.4%) required chest tube insertions. The multivariate logistic regression analysis showed that emphysema, depth of intrapulmonal biopsy path and lesion size were sole effective factors of pneumothorax (OR = 7.991, 1.083, and 0.945 respectively). Lesions with emphysema, depth of intrapulmonal biopsy path and lesion size ≤ 10 mm had higher pneumothorax rates. Eight-seven (18.1%) cases presented pulmonary hemorrhage and twenty-six (5.4%) presented hemoptysis. The multivariate logistic regression analysis showed that depth of intrapulmonal biopsy path, emphysema, and number of pleural needle passes were sole effective factors (OR = 1.143, 0.712, and 0.521, respectively) of pulmonary hemorrhage. In patients with depth > 10 mm or 20 mm, emphysema and number of pleural needle passes had higher hemorrhage rates. The multivariate logistic regression analysis showed that depth of intrapulmonal biopsy path and emphysema were sole effective factors (OR = 1.077, and 0.578, respectively) of hemoptysis. Lesions with depth > 20 mm had higher hemoptysis rate. One case with pulmonary Cryptococcus presented pleural reaction, and three cases had the insistent pain. The total number of severe complications was 15 (3.1%) cases. CONCLUSION: ACNB with EPL was an accurate method for diagnosing pulmonary lesions. ACNB can be safely performed, which reduces the rate of pneumothorax and hemorrhage. Pneumothorax rate was influenced by emphysema, depth of intrapulmonal biopsy path and lesion size ≤ 10 mm. Hemorrhage was related with depth >10 mm or 20 mm, emphysema and number of pleural needle passes. Hemoptysis was related with depth of intrapulmonal biopsy path >20 mm. Later and severe complications should be considered in procedure.

[Clinical analysis of solitary pulmonary nodule measuring less than 10 mm].

BACKGROUND & OBJECTIVE: Lung cancer is a malignant disease with high incidence and mortality, and its prognosis is to large degree determined by early diagnosis and treatment. X-ray chest film and computed tomography (CT) are important means to discover and diagnose lung cancer. But it is difficult to diagnose single pulmonary nodules (SPN) measuring less than 30 mm, especially for those less than 10 mm. This study was to analyze the CT imaging features of SPN measuring less than 10 mm. METHODS: The clinical and radiologic features of 102 cases of pathologically diagnosed SPN measuring less than 10 mm with CT reports were analyzed. The differences between benign and malignant lesions were compared. Imaging features of SPN of lung cancer were summarized. RESULTS: Of the 102 cases of SPN measuring less than 10 mm, 66 (64.7%) were benign lesions, 25 (24.5%) were primary lung cancer, and 11 (10.8%) were metastatic lung cancer. The benign lesions included inflammatory pseudotumor, benign tumor, lung tuberculosis, and so on. Primary lung cancers, including 6 advanced cases (stage IIA and higher stage), had several radiological features different from benign nodules, such as ill-defined margins, speculation, bronchus or vessel involvement. Also, the enhanced CT value was higher in malignant nodules than in benign nodules except inflammation pseudotumor. The SPNs of metastatic lung cancer had similar radiologic features to those of primary lung cancer. CONCLUSION: SPNs measuring less than 10 mm are mainly composed of benign lung tumor and primary lung cancer.