Adult ; Female ; Humans ; Maxillary Sinus Neoplasms ; Middle Aged ; Neoplasms, Muscle Tissue

OBJECTIVETo investigate the treatment of postoperative chyle leak after surgery for digestive malignancies.

METHODSFrom December 2008 to February 2012, in the Sun Yat-sen Memorial Hospital of Sun Yat-sen University, clinical data of 19 patients with chyle leak after digestive system cancer surgery were retrospective analyzed.

RESULTSNineteen cases of chyle leak were all identified between the second and the fourth postoperative day and were all initially managed with conservative treatment including early fasting, parenteral nutrition(PN), 24-hour continuous infusion of somatostatin, and low pressure suction drainage. Eight patients were treated successfully for 6 to 10 days with a significant reduction of the daily drainage volume. Ten patients had enteral nutrition(EN) and their drain tubes were repeatedly washed with 30 ml of compound meglumine diatrizoate injection every day until the drainage volume decreased to 200 ml/day. The time to resolution of chyle leak in these ten patients ranged from 12 to 24 days. One patient had no significant decrease in fluid drainage and developed abdominal distension after one week of conservative treatment. Surgical closure of chyle leak was performed on the 11th postoperative day, abdominal cavity drainage tube was removed on the 4th postoperative day. The patient was discharged home in good condition.

CONCLUSIONMost postoperative chyle leak after surgery for digestive malignancies can be successfully managed with conservative treatment. Somatostatin and the drainage are the main therapeutic approaches. When chyle leak is not resolved with conservative treatment, surgical treatment should be considered to prevent serious complications.

Adult ; Aged ; Anastomotic Leak ; therapy ; Chyle ; Digestive System Neoplasms ; surgery ; Female ; Humans ; Male ; Middle Aged ; Postoperative Complications ; therapy ; Retrospective Studies

OBJECTIVETo screen the specific antibody binding peptides of tuberculosis from phage-displayed random phage display (Ph.D.)-7 peptide libraries with purified IgG from tuberculosis serum, and to provide the basis for the development of serological detection reagent of tuberculosis.

METHODSPurified IgG of tuberculosis serum was used as solid ligand to screen the binding peptide from the Ph.D.-7 peptide library, according to the biopanning process of absorption, elution and amplification. Purified IgG of health people serum was used as the molecule of counter selection during the second and third selection. Phages were enriched after 3 rounds of screening, then 20 single phages separately eluted by IgG of tuberculosis and health people were randomly selected on each direction of the determination plates and amplified. The single chains DNA were extracted as template for sequencing. The combination abilities of selected clones to IgG of tuberculosis and health people were tested by indirect enzyme linked immunosorbent assay (ELISA), and the positive clone was identified. Serum samples, from 47 patients with tuberculosis and 37 healthy people vaccinated with BCG, were verified by positive phage clones using phage-ELISA. The verifying results of 24 serum samples used for panning were separately analyzed statistically.

RESULTSAfter 3 rounds of panning, remarkable enrichment of phages that could specifically bind with target molecules were observed. Single phages were randomly selected for sequencing analysis and 12 sequences were obtained. 12 phage clones with different sequences were amplified and detected with indirect ELISA and single phage H12 showed higher affinity with IgG of tuberculosis (S/N ≥ 2.1) and was identified as the positive clone. It was found that, in indirect ELISA with single Phage H12, the A(450) value of tuberculosis patients (0.105 ± 0.010) was significantly higher than that of healthy individuals (0.070 ± 0.005), and the t value was 2.912 (P < 0.0001). The A(450) value of 12 serum samples of tuberculosis patients and 12 samples of health individuals used for panning were 0.144 ± 0.016 and 0.052 ± 0.004, and the t value was 5.69 (P < 0.0001).

CONCLUSIONBy using phage-displayed random peptide libraries, we obtained the specific antibody binding peptides of tuberculosis, which showed specific binding activity with IgG of tuberculosis. It can provide a basis for the establishment of a new serological detection method of tuberculosis with peptide.

Adult ; Case-Control Studies ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; Molecular Sequence Data ; Mycobacterium tuberculosis ; immunology ; Peptide Library ; Peptides ; immunology ; Tuberculosis ; diagnosis ; immunology ; microbiology ; Young Adult

OBJECTIVETo compare the Epstein-Barr virus (EBV) infection rates and the frequencies of wt-LMP1 and del-LMP1 EBV variants detected singly or dually among the four types of nasopharyngeal carcinoma (NPC) and to illustrate the possible role of del-LMP1 gene in nasopharyngeal carcinogenesis.

METHODSEBER in situ hybridization was performed in 117 NPCs, including 48 non-keratinizing carcinomas (NKCs), 25 keratinizing squamous cell carcinomas (KSCCs), 5 adenosquamous carcinomas (ASCs), 6 mucoepidermoid carcinomas (MECs) and 33 adenocarcinomas (ACs). Nested PCR for demonstration of EBV LMP1 gene was performed on the tissue samples collected from 99 EBER-positive carcinoma cases and the peripheral blood mononuclear cells (PBMCs) of 53 healthy adults (HAs).

RESULTSAs indicated by EBER in-situ hybridization, the EBV infection rates in both of 48 NKCs and 25 KSCCs were 100%; and the infection rates of 11 ASCs/MECs and 33 ACs were 9/11 and 51.5% (17/33), respectively. Worthy to note was that most of the NKC cells were EBER-positive while only a small number of EBER-positive neoplastic cells could be found in 17 ACs. The percentage of del-LMP1 EBV variant detected singly in NKCs (85.4%, 41/48) was not only significantly higher than that in PBMCs of 46 HAs (8.7%, 4/46) but also significantly higher than those detected in KSCCs (16.0%, 4/25). The dual infection rate of wt-LMP1 and del-LMP1 variants detected in KSCCs (56.0%, 14/25) was significantly higher than that of NKCs (12.5%, 6/48). The majority of the EBV detected in AC tissues (12/17) and HAs' PBMCs (34/46, 73.7%) were of dual wt-LMP1 and del-LMP1 variants.

CONCLUSIONSThe EBV infection rates are significantly different among 3 major histological categories, namely, NKC/KSCC, ASC/MEC and AC. Though NKCs and KSCCs are always consistently associated with EBV, the single del-LMP1 EBV variant detected in NKCs is predominant over that in KSCCs and most of the KSCCs contain dual wt-LMP1 and del-LMP1 EBV variants. The EBV of the del-LMP1 variant might play a crucial role in carcinogenesis of NKC.

Adult ; Epstein-Barr Virus Infections ; epidemiology ; Female ; Gene Deletion ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; etiology ; virology ; Viral Matrix Proteins ; genetics

OBJECTIVETo investigate the clinicopathologic characteristics of primary nasopharyngeal adenoid cystic carcinoma (NPACC) and its relation to Epstein-Barr virus (EBV) infection in Guangzhou where is a high-incidence area of EBV-associated nasopharyngeal carcinoma (NPC).

METHODS17 cases of NPACC with clinical record and biopsy samples were collected in Guangzhou and their clinical manifestations were reviewed. Besides HE, Alcian blue and PAS, LSAB immunohistochemistry was performed for detecting the expression of a variety of epithelial markers, CD21 and EBV encoded LMP1. EBV encoded early RNAs (EBER) was detected by using in-situ hybridization. Nested PCR was applied for studying the presence of EBV W-fragment in tissues.

RESULTSThe ratio of male to female was 7:10. The patients' age ranged from 30 to 63 years, and the median age was 46 years. 14 out of 17 tumors showed markedly local aggressive growth, presenting as T3 or T4. However, only 1 patient had metastasis of an ipsilateral cervical lymph node. The majority of neoplastic cells were basal-cell like in shape and with scanty cytoplasm and a deeply stained nucleus. Intercellular hyaline or mucinous substance was always present in between the carcinoma cells. Cribriform structure formed by the neoplastic cells could be found in 16 out of these 17 biopsies. The NPACC always express the wide-spectrum cytokeratin and the epithelium membrane antigen. Only a few or a small number of carcinoma cells showed nuclear EBER-signals in 9 cases (9/17). Concurrently, these 9 NPACCs showed a 192 bp W-fragment positive band on electrophoresis gel by nested PCR. LMP1 expression had been found in 5 out of the 9 NPACCs (55.6%) accompanying with EBER-positive carcinoma cells. The EBER-positive infiltrating lymphocytes could also be found in the stroma of 3 out of the 9 EBER-stained NPACC slides. All the tumor cells, including the EBER-positive cell of the 17 NPACCs showed no CD21 expression.

CONCLUSIONSThe female is predominant over the male in development of NPACC, which often accompanied with a markedly invasive capacity at the nasopharynx and its neighboring sites. Only a small number of tumor cells, nearly a half of the studied cases were infected with EBV. Therefore, it's postulated that there seems no close relation present between NPACC and EBV infection.

Adult ; Carcinoma, Adenoid Cystic ; etiology ; pathology ; Epstein-Barr Virus Infections ; complications ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; etiology ; pathology ; RNA, Viral ; analysis ; Viral Matrix Proteins ; analysis

Adult ; Epstein-Barr Virus Infections ; complications ; Humans ; Leukocytes, Mononuclear ; virology ; Nasopharyngeal Neoplasms ; pathology ; virology ; Viral Matrix Proteins ; genetics

OBJECTIVE: To examine the correlation of CCBE1 expression in adjacent tissues of tongue squamous cell carcinoma (TSCC) with pericancerous lymphatic vessel proliferation, cervical lymph node metastasis and survival outcomes of the patients. METHODS: Lymphatic vessel density was quantified in pericancerous tissue sections of 44 cases of cT_1-2N₀ TSCC using D2-40 as the lymphatic vessel endothelial marker for calibration and counting of the lymphatic vessels. Of these 44 cases, 22 showed a relatively low lymphatic vessel density (group A) and the other 22 had a high lymphatic vessel density (group B), and the expression levels of CCBE1 in the adjacent tissues determined using immunohistochemistry, immunofluorescence assay and Western blotting were compared between the two groups. The expression level of CCBE1 was also measured in another 90 patients with TSCC using immunohistochemistry, and all the patients were followed up for their survival outcomes. RESULTS: Immunohistochemistry and Western blotting showed a significantly lower rate of high CCBE1 expression in group A than in group B (P < 0.05). Immunofluorescence assay showed co-localization of CCBE1 and D2-40 in the adjacent tissues of TSCC. In the 90 TSCC patients with complete follow-up data, a high expression of CCBE1 was found to correlate with lymph node metastasis and a poor 5-year survival outcomes of the patients (P < 0.05). CONCLUSION A high expression of CCBE1 in the adjacent tissues of TSCC is closely related with pericancerous lymphatic vessel proliferation, cervical lymph node metastasis and a poor 5-year survival of the patients, suggesting the value of CCBE1 as a potential prognostic predictor for TSCC.
Humans ; Tongue Neoplasms/pathology* ; Carcinoma, Squamous Cell/metabolism* ; Lymphatic Metastasis ; Prognosis ; Lymphatic Vessels/pathology* ; Cell Proliferation ; Tongue/pathology* ; Calcium-Binding Proteins/metabolism* ; Tumor Suppressor Proteins/metabolism*

Adult ; Embryonic Development ; Female ; Fetal Growth Retardation ; etiology ; physiopathology ; Gestational Age ; Humans ; Pregnancy ; Pregnancy Trimester, First

OBJECTIVETo detect the sequence variations frequently found within the N- and C-terminal regions of Epstein-Barr virus (EBV) LMP1 gene in nasopharyngeal carcinoma (NPC) and to study the underlying mechanisms.

METHODSFresh tumor tissues were sampled from 63 patients with untreated NPC encountered in Affiliated Tumor Hospital of Sun Yat-sen University, Guangzhou. The N-terminal region of EBV LMP1 gene was amplified with nested polymerase chain reaction (PCR), followed by XhoI enzyme digestion. Nested PCR was also employed to detect the 30 base pairs deletion within the C-terminal region. Four-colored fluorescence terminator sequencing method was applied for bi-directional solid-phase sequencing of the 8 representative PCR products in 4 cases of NPC. The DNA sequence within the N- and C-terminal regions of LMP1 gene was then analyzed.

RESULTSThere were 4 patterns of sequence variations, namely, wt-XhoI/wt-LMP1 (4 cases, 6.3%), wt-XhoI and XhoI-loss/del-LMP1 (4 cases, 6.3%), wt-XhoI/del-LMP1 (5 cases, 7.9%) and XhoI-loss/del-LMP1 (50 cases, 79.5%), detected in the 63 studied cases. Sequence analysis showed that the EBV LMP1 gene had underwent non-synonymous and synonymous substitutions, as compared with the prototype of B95-8 cells. The ratio of non-synonymous to synonymous substitutions was 2.25.

CONCLUSIONSXhoI-loss/del-LMP1 is the predominant sequence variation pattern of EBV LMP1 gene in NPC from Guangzhou. The XhoI-loss variation seems to develop on top of del-LMP1. When compared with the EBV LMP1 gene in peripheral blood B-lymphocytes of virus carriers and in preinvasive epithelial lesions (reported previously), it is likely that the sequence variation patterns of LMP1 gene may represent 4 different phases of intrahost evolution of EBV during nasopharyngeal carcinogenesis.

Adult ; Aged ; Base Sequence ; DNA, Viral ; genetics ; Deoxyribonucleases, Type II Site-Specific ; genetics ; Female ; Gene Deletion ; Genetic Variation ; Herpesvirus 4, Human ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Nasopharyngeal Neoplasms ; virology ; Point Mutation ; Sequence Analysis, DNA ; Viral Matrix Proteins ; genetics

OBJECTIVETo investigate the risk factors of intrauterine hepatitis B virus (HBV) infection and the impact of HBV DNA on the infection.

METHODSThe serum levels of HBsAg, HbsAb, HBeAg, HBeAb, HBcAb and HBV DNA were determined in blood samples from 230 HBsAg-positive pregnant women and their newborns by enzyme-linked immunosorbent assay (ELISA) and fluorescence quantitative PCR (FQ-PCR), respectively. The newborns acquiring HBV infection via intrauterine transmission were selected as the case group and others as the control group. The risk factors for intrauterine HBV infection were analyzed by non-conditional logistic regression model.

RESULTSSix infants were found to be HBsAg-positive, and 18 HBV DNA-positive, and 3 of them were positive for both HBsAg and HBV DNA. The rate of intrauterine HBV infection was 9.6% (22/230). The grade of HBV DNA level was identified as the only risk factor of intrauterine HBV infection by non-conditional logistic regression model, with odds ratio (OR) of 1.57 (95% confidence interval 1.12-2.21). Of the 119 pregnant women positive for HBV DNA, 18 were diagnosed as having intrauterine HBV infection, and the likeliness of the infection significantly increased for a maternal serum HBV DNA level > or =10(7) copies/ml (chi(2)=7.92, P<0.05).

CONCLUSIONThe grade of serum HBV DNA level is the predominant risk factor for intrauterine HBV infection in pregnant women, and for those with serum HBV DNA lever > or =10(7) copies/ml, the chance for intrauterine HBV infection can be significantly increased.

DNA, Viral ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Hepatitis B ; transmission ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Logistic Models ; Odds Ratio ; Polymerase Chain Reaction ; Pregnancy ; Pregnancy Complications, Infectious ; blood ; virology ; Risk Factors