Evaluation and comparison of the diagnostic performance of routine blood tests in predicting liver fibrosis in chronic hepatitis B infection.

Background & aims: Biopsy is the gold standard for staging liver fibrosis, but it may be accompanied by complications. As an alternative, non-invasive markers such as transient elastography (for liver fibrosis) and certain combinations of routine blood markers (liver function tests, full blood count) have been developed although their clinical significance remains controversial. Here, we compare the diagnostic values of non-invasive markers for liver fibrosis in patients with chronic hepatitis B infection. Methods: Transient elastography and routine laboratory tests were performed in 196 patients. Diagnostic performances were compared and were assessed based on the area under the curve (AUC) of a receiver operating characteristic (ROC) analysis. Results: Elevated GGT to platelet ratio (GPR), the fibrosis index FIB-4 [based on age, AST, platelets and ALT], platelet to lymphocyte ratio (PLR) and total bilirubin were independent predictors of liver stiffness defined by transient elastography (all P < 0.001). The AUCs of GPR in predicting both advanced fibrosis and cirrhosis were significantly larger than that of FIB-4 (P = 0.037 and P = 0.008, respectively) and AST-to-platelet ratio index (APRI) (P = 0.008 and P = 0.005). FIB-4, APRI and red cell volume distribution width-to-platelet ratio (RPR) had similar diagnostic values in discriminating different levels of liver fibrosis. Conclusions: GPR showed the best diagnostic value and RPR and PLR are easily available and inexpensive markers in evaluating fibrosis and cirrhosis. The diagnostic values of these laboratory markers are useful in diagnosing advanced fibrosis or cirrhosis, and in confirming the different levels of liver fibrosis.

MiR-940 inhibits the progression of NSCLC by targeting FAM83F.

OBJECTIVE: We investigate whether miR-940 could target family sequence similarity 83 member F (FAM83F) and further inhibit the progression of non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: The expression levels of miR-940 and FAM83F in tumor tissues and paracancerous tissues of 72 NSCLC patients were detected through quantitative real time-polymerase chain reaction (qRT-PCR). The relationship between their expression levels, tumor size, and prognosis of NSCLC was analyzed. Transfection plasmids were constructed to knockdown or overexpress miR-940 in H1299 cells (inhibitor group) and SK-MES-1 cells (mimic group). The viability of H1299 cells and SK-MES-1 cells was evaluated using cell counting kit-8 (CCK-8) assay after transfection. The combination of miR-940 and Ago2 was confirmed by RNA immunoprecipitation (RIP) experiment. The binding condition of miR-940 in FAM83F-WT and FAM83F-MUT groups was verified by luciferase reporter gene assay. RESULTS: MiR-940 expression was noticeably decreased, while FAM83F expression was distinctly upregulated in NSCLC tissues than that of paracancerous tissues. The overall survival rate of NSCLC patients with highly-expressed miR-940 was significantly higher than those with lowly-expressed miR-940. Besides, miR-940 level was negatively correlated with tumor stage and size of NSCLC patients. Knockdown of miR-940 evidently enhanced the activity of H1299 cells, while overexpression of miR-940 decreased the viability of SK-MES-1 cells. In addition, miR-940 was confirmed to combine with FAM83F. Luciferase activity of cells co-transfected with FAM83F-WT and miR-940 mimic was significantly decreased. CONCLUSIONS: MiR-940 inhibited the proliferation of cancer cells by targeting FAM83F and further restrained the progression of NSCLC.

A new route of transendocardial stem cell injection: from femoral vein to left ventricle through atrial septa.

OBJECTIVE: To evaluate the feasibility and safety of a new route of transendocardial stem cell injection. MATERIALS AND METHODS: Atrial septal puncture was performed in six young male pigs, and then a 6F syringe was passed through the puncture to reach the left atrium. Meanwhile, a guide wire was inserted into the left ventricle through the femoral artery, and echocardiography was used to confirm the relative position of the guide wire with the syringe. RESULTS: After atrial puncture through femoral vein, the syringe could reach the left atrium and finally the left ventricle. Echocardiography confirmed that both the guide wire and the syringe were located in the left ventricle. The diameter of atrial septa puncture and the diameter of the syringe implantation were 4.1 ± 0.5 mm and 8.4 ± 0.7 mm, respectively. But there is no difference in Left ventricle end-systolic dimension (LVES), left ventricle end-diastolic dimension (LVED) and left ventricle ejection fraction (LVEF) before surgery, after atrial septa puncture, after syringe implantation or one month after surgery. CONCLUSIONS: It is feasible to perform transendocardial stem cell injection by 6F syringe inserted through femoral vein. The surgery may cause atrial septa tearing but does not jeopardize myocardial function.

Tumor development is associated with decrease of TET gene expression and 5-methylcytosine hydroxylation.

The TET (ten-eleven translocation) family of α-ketoglutarate (α-KG)-dependent dioxygenases catalyzes the sequential oxidation of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine and 5-carboxylcytosine, leading to eventual DNA demethylation. The TET2 gene is a bona fide tumor suppressor frequently mutated in leukemia, and TET enzyme activity is inhibited in IDH1/2-mutated tumors by the oncometabolite 2-hydroxyglutarate, an antagonist of α-KG, linking 5mC oxidation to cancer development. We report here that the levels of 5hmC are dramatically reduced in human breast, liver, lung, pancreatic and prostate cancers when compared with the matched surrounding normal tissues. Associated with the 5hmC decrease is the substantial reduction of the expression of all three TET genes, revealing a possible mechanism for the reduced 5hmC in cancer cells. The decrease of 5hmC was also observed during tumor development in different genetically engineered mouse models. Together, our results identify 5hmC as a biomarker whose decrease is broadly and tightly associated with tumor development.

Intraosseous venous malformations of the facial bone: a retrospective study in 11 patients.

OBJECTIVES: To investigate the clinical symptoms, computed tomography (CT) features and treatments of intraosseous venous malformations (IVM) that occur in the facial bone. METHODS AND RESULTS: Eleven patients with facial IVM were treated with two surgical techniques, excision (n = 4) or curettage (n = 7). No recurrence was encountered at follow-up (45.8 ± 16.0 months). Postoperative deformities were left in two paediatric patients who were treated with excision. CONCLUSIONS: The diagnosis of IVM can be difficult and is mainly based on clinical symptoms and CT features. IVM should be differentiated from other lesions, including ameloblastoma, odontogenic cysts, osteosarcoma, aneurysmal bone cysts and arteriovenous malformations, among others. Conventional block biopsy should be replaced by fine needle aspiration cytology for further diagnosis. Curettage is a more appropriate method for IVM compared with excessive en-bloc osteotomy, while transosseous embolo-sclerotherapy may be a promising alternative method. Finally, the terminological confusion between 'intraosseous haemangioma' and 'intraosseous venous malformation' should be avoided according to the binary classification.

Sclerotherapy of voluminous venous malformation in head and neck with absolute ethanol under digital subtraction angiography guidance.

OBJECTIVES: Venous malformation (VM) is the most common symptomatic low-flow vascular malformation, which predominantly occurs in the head and neck region. The aim of this paper was to evaluate the results of endovascular sclerotherapy of voluminous VM, when the lesion is either >or=15 cm in maximum diameter or the lesion invades more than one anatomical space, in the head and neck region using absolute ethanol under digital subtraction angiography (DSA) guidance. METHODS: A total of 23 patients with head and neck VMs between October 2005 and December 2008 were retrospectively reviewed. All patients received direct puncture ethanol sclerotherapy under DSA guidance. Follow-up assessments were performed at 3-25 months after therapies were completed, and complications were reported in some cases. RESULTS: All patients were satisfied with the results of therapy. Seventeen patients (73.9%) achieved excellent responses and six patients (26.1%) achieved good responses in magnetic resonance imaging assessments. Minor complications developed during the procedures, all of which were successfully managed with full recovery during follow-ups. Serious complications such as acute pulmonary hypertension, cardiovascular collapse and pulmonary embolism were not encountered. CONCLUSION: It is concluded that sclerotherapy with absolute ethanol under DSA guidance is an important alternative therapy for voluminous and extensive VM, as the procedure is reasonably safe and offers good therapeutic results.

Angiosarcomas of the head and neck: a clinico-immunohistochemical study of 8 consecutive patients.

Angiosarcoma, also known as malignant hemangioendothelioma, is a rare and aggressive malignant vascular tumour arising from endothelial cells, which accounts for approximately 10% of soft tissue sarcomas in the head and neck. Between October 1996 and July 2008, 10 patients were diagnosed with angiosarcomas (AS) in the head and neck region, 8 of whom were included in this study (there were 7 high-grade and 1 low-grade lesions). AS were characteristically positive for vascular markers such as CD31, CD34, and factor VIII. CD31 was thought to be the most sensitive and specific marker for AS and was positive in 6 patients. Six patients were treated surgically with or without postoperative radiotherapy and/or chemotherapy. Two patients had large and extensive lesions that were considered to be inoperable, they were given palliative chemotherapy and/or radiotherapy. Of the 8 patients reviewed in this study, 5 died of local recurrence or distant metastasis with a survival time of 8-19 months, 2 patients are alive with disease and 1 patient is free of disease. The predeliction for local recurrence and distant metastasis and the high-grade characteristics of this tumour seemed to be correlated to the poor prognosis, although the small number of patients prevented statistical analysis.

Hepatitis B virus X protein induces hypermethylation of p16(INK4A) promoter via DNA methyltransferases in the early stage of HBV-associated hepatocarcinogenesis.

The aim of the present study was to authenticate the involvement of DNA methyltransferases (DNMTs) and methyl-CpG binding domain protein 2 (MBD2) in the process of HBx induced p16(INK4A) promoter hypermethylation in HBV-related hepatocellular carcinoma (HCC) and their corresponding noncancerous liver tissues. Eighty-eight fresh tissue specimens of surgically resected HBV-associated HCC and their corresponding noncancerous liver tissues were studied. The methylation status of the p16(INK4A) promoter was determined by methylation-specific polymerase chain reaction (MSP). Reverse transcription and real-time polymerase chain reaction (RT-PCR) showed the expression of DNMTs, MBD2 and HBx. Western blot and immunohistochemistry were used for the protein analysis of HBx, DNMT1, DNMT3A and P16. Tissue HBV-DNA levels were determined by RT-PCR. HBV genotype was examined by nested PCR and restriction fragment length polymorphism (RFLP). In the corresponding noncancerous liver tissues, higher HBx expression was associated with the hypermethylation of the p16(INK4A) promoter. HBx was positively correlated with the DNMT1 and DNMT3A at both the mRNA and protein level. Furthermore, HBx, DNMT1 and DNMT3A protein expression were negatively correlated with p16 protein expression. In HCC tissues, HBx was positively correlated with DNMT1 and DNMT3A at both mRNA and protein level, but HBx expression did not correlate with hypermethylation of the p16(INK4A) promoter or p16 protein expression. The methylation status of the p16(INK4A) promoter did not correlate with clinicopathological characteristics. DNMT1 and DNMT3A may play important roles in the process of HBx inducing hypermethylation of the p16(INK4A) promoter in the early stages of HBV-associated HCC. HBx-DNMTs-p16(INK4A) promoter hypermethylation may constitute a mechanism for tumorigenesis during HBV-associated hepatocarcinogenesis.

Malignant solitary fibrous tumors of the head and neck: a clinicopathological study of nine consecutive patients.

Malignant solitary fibrous tumor (MSFT) is one of the angiosarcomas arising from or resembling the components of blood vessels. Because of its rarity, the diagnosis is difficult. Between 1998 and 2008, 24 patients were diagnosed as having hemangiopericytomas (16) and solitary fibrous tumors (8). Nine of them were confirmed to be MSFT and treated at the Department of Oral and Maxillofacial Surgery, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine. All patients were treated with surgery as the primary modality. Eight patients received postoperative adjuvant radiation therapy. The average follow-up duration was 4.6 years (range: 6 months-10 years). Of the nine patients reviewed in this study, three died of locoregional or distant metastasis, six survived 6 months-6 years (four survived over 4 years). The size of primary tumor, the number of mitoses per 10 high powered fields and cervical lymph node metastasis seemed to be correlated with poor prognosis in MSFT patients, although the small number of patients did not permit the performance of statistical analysis.

Clonal analysis of gastric carcinoma and precancerous lesions and its relation to Ki-67 protein expression.

The pathogenesis of intestinal carcinoma is characterized as progressing through multiple steps, which begin with atrophic gastritis followed by intestinal metaplasia, dysplasia and carcinoma. However, the clonal status of gastric precancerous lesions and its association with proliferative kinetics have not been fully understood. In this study, gastric lesions and normal epithelial cells were isolated from formalin-fixed paraffin embedded tissues using a laser capture microdissection (LCM) system, the clonality was analyzed with human androgen receptor gene (HUMARA) polymerase chain reaction (PCR), and the PCR products were examined using Applied Biosystems 3730 DNA Analyzer. The relationship between the clonal status and Ki-67 protein expression was also investigated. Ki-67 was detected by two-step immunohistochemical staining. 5/32 intestinal metaplasia lesions, 10/45 low grade intraepithelial neoplasia, 25/36 high grade intraepithelial neoplasia and 20/20 intestinal gastric carcinoma were of monoclonal origin. Similar to monoclonal inactivation, the expression rate of Ki-67 also increased along the multi-step gastric carcinogenesis. Clonal status was associated with the expression rate of Ki-67 to a certain extent, which may be useful in assessing susceptibility to gastric carcinoma. Key words: gastric carcinoma; precancerous lesion; clonal analysis; Ki-67.

Infant ectopic cervical thymus in submandibular region.

Sporadic cases have been reported of ectopic thymic tissue formed along the path of embryologic descent from the mandibular region to the mediastinum, usually manifesting as an asymptomatic mass. Here is reported the case of an 8-month-old boy with a tender palpable mass in the right upper lateral neck. Preoperative posteroanterior chest radiograph revealed normal structures in the mediastinum superior including the thymus. Magnetic resonance imaging showed a 4-cm x 4-cm soft-tissue mass in the left submandibular region. Surgical resection was performed and histopathologic examination showed that the mass was composed of thymic lymphoid tissue and epithelial cells. Immunohistochemical features included positive expression of LCA, CKpan, EMA, CD20 and CD43 antibodies. The clinical 14-month follow up was negative and the child was growing normally after operation. Ectopic thymus in the submandibular region is uncommon; surgical treatment is the definitive means of pathological diagnosis. Prior to surgery, the presence of a mediastinal thymus should be confirmed to prevent the risk of a total thymectomy.

Clonality analysis of intraductal proliferative lesions using the human androgen receptor assay.

Recently, it is accepted that invasive breast carcinoma is of monoclonal origin. Ductal intraepithelial neoplasia (DIN) may progress toward invasive carcinoma with an increased risk. However, it is not fully understood whether DIN is polyclonal or monoclonal. In this current study, we detected clonal origin of DIN using x-inactivation at the human androgen receptor (HUMARA) locus. Lesional and normal breast gland cells were microdissected from paraffin-embedded tissues using a laser capture microdissection system. Genomic DNA was extracted. After digestion by restriction enzyme Hpa II, the HUMARA exon1 was amplified by a fluorescent nested-PCR procedure and the PCR products were separated on DNA sequencer and analyzed the fluorescent intensity of the two HUMARA alleles. DNA from 88 of 101(87%) patients was able to be amplified at the HUMARA locus and 68 of them (77.3%) were heterozygous and informative. 9/12 usual ductal hyperplasia (UDH) and 5/18 DIN 1A showed a polyclonal inactivation. 3/12 UDH, 13/18 DIN 1A, 28/28 DIN 1B, 10/10 carcinoma in situ are of monoclonal origin. Taken together, DIN 1A, 1B and carcinoma in situ, are monoclonal and DIN 1, but not UDH, represents the obligate and direct precursor of DCIS.

Carotid artery resection and reconstruction: clinical experience of 28 consecutive cases.

The aim of this study was to analyse the experience at a single institution in carotid artery resection with or without reconstruction performed as part of an oncological procedure or emergency haemostasis. A total of 28 patients were included in this retrospective study; 17 underwent ligation or resection of the carotid artery, and 11 underwent reconstruction of the carotid artery. The perioperative complications and surgical outcomes were recorded and analysed. Of the 17 patients with ligation or resection of the carotid artery, 4 developed neurologic deficit within 2 weeks postoperatively. Three patients with malignant tumours died 1 month (1) and 4 months (2) postoperatively. Of the 11 patients undergoing carotid reconstruction, no major cerebral complications were noted after operation. Colour Doppler showed patent vascular graft 1 year postoperatively in nine patients. Due to the higher complication rates both in short and long term with ligation or resection of the carotid artery, resection and revascularization of the carotid artery is advocated for patients with carotid artery involvement when possible.

Detection of serum Cyfra 21-1 in patients with primary oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is the most common malignant tumour in the oral and maxillofacial region, and has a poor prognosis. Cyfra 21-1 is a useful tumour marker for squamous cell carcinoma, but the clinical value of Cyfra 21-1 in OSCC has not been confirmed. In order to investigate the diagnostic and prognostic value of serum Cyfra 21-1 in primary OSCC patients, the preoperative serum Cyfra 21-1 concentration of 100 OSCC patients and 56 healthy subjects was detected by enzyme-linked immunosorbent assay (ELISA). The cut-off value was calculated with a receiver operating characteristic (ROC) curve, and prognostic analysis was performed using the Kaplan-Meier method and Cox regression models. The preoperative serum Cyfra 21-1 concentration in OSCC patients (1.18+/-1.20 microg/L) was significantly higher (t=6.585, P<0.001) than that in healthy subjects (0.40+/-0.16 microg/L). With a cut-off value of 0.65 microg/L, the diagnostic sensitivity and specificity was 0.570 and 0.964, respectively. There was significant correlation with tumour recurrence and survival rate: the higher the serum Cyfra 21-1; the higher the tumour recurrence rate and lower the survival rate. Serum Cyfra 21-1 was an independent prognostic factor for OSCC using univariate and multivariate Cox models.

Gene expression profile of transgenic mouse kidney reveals pathogenesis of hepatitis B virus associated nephropathy.

Hepatitis B virus (HBV)-associated nephritis has been reported worldwide. Immune complex deposition has been accepted as its pathogenesis, although the association between the presence of local HBV DNA and viral antigen and the development of nephritis remains controversial. To understand better the roles played by HBV protein expression in the kidney, the global gene expression profile was studied in the kidney tissue of a lineage of HBV transgenic mouse (#59). The mice expressed HBsAg in serum, and HBsAg and HBcAg in liver and kidney, but without virus replication. Full-length HBV genome (adr subtype, C genotype) isolated from a chronic HBV carrier was used to establish the transgenic mice #59. Similarly manipulated mice that did not express HBV viral antigens served as controls. Southern blotting, hybridization with HBV probe, and immuno-histochemical staining were used to study HBV gene expression. mRNA extracted from the kidney tissue was analyzed using Affymetrix microarrays. HBsAg and HBcAg were located mainly in the cytoplasm of tubular epithelium. Altogether 520 genes were "up-regulated" more than twofold and 76 genes "down-regulated" more than twofold in the kidney. The complement activation, blood coagulation, and acute-phase response genes were markedly "up-regulated". Compared to the controls, the level of serum C3 protein was decreased in #59 mice, while the level of C3 protein from kidney extract was increased. Results indicate that expression of HBsAg and HBcAg in tubular epithelial cells of the kidney per se can up-regulate complement-mediated inflammatory gene pathways, in addition to immune complex formation.

[Reconstruction of the common carotid artery with freeze dried arterial allografts in goats].

OBJECTIVE: To observe the patency rate and histopathologic changes at different times after implantation of freeze dried arterial allografts. METHODS: 4 healthy adult goats were used as the experimental models. A segment of the common carotid artery 2 cm in length was surgically removed and bridged with freeze-dried graft of common carotid artery from another goat. The eight necks of 4 animals were divided into A and B group randomly, each containing 4 arteries. In group A, both cut ends of the carotid artery were anastomosed end to end with the arterial allograft. In group B, the proximal end of the carotid artery was anastomosed end to side with the allograft. The patency of the graft was monitored with Laser Doppler Flowmeter within one week and carotid angiography was performed in selected animal. The allografts with anastomoses were harvested at 1 week, 2 weeks, 3 months and 6 months after implantation, respectively. The surgical fresh specimens were prepared and subject to histopathologic evaluation under light microscope, scanning electron microscope and transmission electron microscope. RESULTS: In Group A, 2 grafts were patent at 1 week and 2 weeks, respectively. One graft was not patent at 3 months, one was largely occluded at 6 months, only 1/4 part of the graft was patent. In Group B, 3 grafts were patent at 1 week, 2 weeks, 3 months, respectively. One graft was occluded at 6 months. There was no significant difference in patency rate between the two groups. Histopathologic examinations showed that in freezed dried arterial homograft, the endothelial cells were lost but the matrix was complete. One week after implantation, mononuclear macrophages were infiltrated into the wall in large amount and adventitia of loose connective tissues was formed. After 2 weeks of implantation, intimia was noted, intimal hyperplasia was also noted at the anastomotic sites and migrated toward the central portions of the graft. At 3 months, the homograft displayed a complete endothelial cell coverage with only small areas of fresh fibrin and endothelial disruption. The rejection reaction was diminished prominently. The medial matrix was largely substituted by proliferation of smooth muscle cells, the homograft was near normal in structure. CONCLUSION: Reconstruction of carotid artery defect with freeze dried arterial homografts was feasible. The rejection reaction was not significant. If the endothelial lining does not regenerate within 3 months, long term graft patency is hardly achieved.

[Reconstruction of the common carotid artery with autogenous external jugular vein graft in goats].

OBJECTIVE: To establish a method for reconstruction of carotid artery defect with autogenous graft of external jugular vein, to investigate the patency rate and histopathologic changes at different times after implantation. METHODS: 3 healthy adult goats were used as the experimental models. A segment of 2 cm autogenous external jugular vein was used to bridge a surgical defect of the common carotid artery (2 cm long). The six necks of the three animals were divided into A and B group randomly. In Group A, both cut ends of the artery were anastomosed end to end with the venous graft. In Group B, the proximal end of the artery was anastomosed end to side with the venous graft. The patency rate was recorded at different times. The venous grafts with anastomotic sites were harvested at 2 weeks, 3 months and 6 months after implantation, respectively, for histopathologic examinations. RESULTS: The grafts in Group A were patent at 2 weeks and 3 months, but occluded at 6 months. The grafts in Group B were patent at 2 weeks and 6 months, but not patent at 3 months. Chi-square test demonstrated insignificant difference in patency rate between Group A and Group B. Histopathologic examinations showed intimal hyperplasia and venous arterialization in venous autografts. The venous wall was loose and edematous, intimal and subintimal hyperplasia was prominent at 2 weeks. Proliferation and migration of smooth muscle cell was found at 3 months after implantation. At 6 months, the venous media was predominantly composed of smooth muscle. Intimal hyperplasia was most prominent adjacent to the anastomosis. CONCLUSION: Reconstruction of the common carotid artery with autogenous graft of external jugular vein had a higher patency rate. The main histopathologic changes after implantation is intimal hyperplasia and venous arterialization. Intimal hyperplasia is the result of proliferation and migration of host arterial smooth muscle cell, which passes through the anastomosis to the intima of the vein graft.

The identification and toxicity of the microcystin in the waterbloom obtained from an eutrophied lake.

The water of "J" lake has been seriously eutrophied; concentration of total nitrogen (TN), total phosphorus (TP) and chlorophyll a were all far above the 3rd level of the National Standard of Ground Water of China. The concentration of microcystin (MCYST) of the water at one site (M) was 1865 micrograms/l. There were 2.36 micrograms MCYST-LR per mg dry waterbloom powder.