Volume 120  ·   Issue 18  ·  2007

Abstract：Background Aberrant DNA methylation plays a key role in human carcinogenesis. 5-aza-2'-deoxycytidine inhibits DNA methylation and induces the expression of genes putatively silenced by promoter methylation in vitro. There are few studies of the biological and clinical significance of 5-aza-2'-deoxycytidine in human hepatocellular carcinoma. This study explored the mechanism of 5-aza-2'-deoxycytidine targeting transcriptional repressor complexes affecting global gene expression in hepatocellular carcinoma cell line.Methods High density oligonucleotide gene expression microarrays were used to examine the effects of 5-aza-2'-deoxycytidine treatments on human hepatocellular carcinoma cell line SMMC-7721. The 5' ends of the genes upregulated or downregulated in this manner were compared with BLAST database to determine whether they might have promoter CpG islands. Flow cytometry was used to detect stages of the cell cycle and apoptosis of SMMC-7721 after being treated with 5-aza-2'-deoxycytidine.Results Data obtained 3 days after 4 days of treatment with 5-aza-2'-deoxycytidine showed that more genes were induced in tumorigenic cells including genes that function in cell proliferation, differentiation, regulation of transcription,and cytokine signalling. Approximately 30% of induced genes did not have CpG islands within their 5' regions, suggesting that some genes activated by 5-aza-2'-deoxycytidine may not result from the direct inhibition of promoter methylation.This phenomenon may contribute to a number of upregulated genes involving regulation of transcription in the treated cell. Results showed that 100 μmol/L 5-aza-2'-deoxycytidine blocked cell cycle at S/G2-M phase increasing rate of apoptosis. Notably, we found differential expression of molecular action in the methylation although DNA methyltransferases did not show significant difference in the treated cell line.Conclusion 5-aza-2'-deoxycytidine could restore some silenced genes expression independently of DNA methylation inhibition and expression of DNA methyltransferases。

Abstract：Background Suicide gene therapy is a widely used molecular treatment for head and neck cancer. In this study, we try to use the method of homogenous recombination in bacteria to clone thymidine kinase gene (tk)-a kind of suicide gene to adenovirus backbone vectors for the construction of replication-defective adenoviruses.Methods pAdTrack-CMV/tk was constructed through subclone of a restriction endonuclease fragment including thymidine kinase gene from plasmid pCMV-tk to another plasmid pAdTrack-CMV, and then co-transfected with supercoiled pAdEasy-1, which was an adenoviral backbone vector except for deletions of E1 and E3, to competent E.coli BJ5183 for homogenous recombination using electroporation procedure. With the same method, pAdTrack-CMV was also co-transformed with pAdEasy-1 for homogenous recombination in BJ5183. Identified with restriction endonuclease Pacl and polymerase chain reaction (PCR), plasmids pAd-GFP/tk and pAd-GFP were successfully constructed. Each of them was digested with Pacl and sequently transfected into human embryo kidney 293 cells (HEK293) using Lipofectamine 2000.Results Comet-like adenovirus-producing foci of Ad-GFP/tk and Ad-GFP were observed after 5 to 7 days of cell culture.After twelve days of packaging, the replication-defective adenoviruses were collected. Identified with PCR, thymidine kinase gene was successfully constructed into Ad-GFP/tk.Conclusion The replication-defective adenoviruses containing thymidine kinase can be constructed more easily by homogenous recombination in bacteria than conventional techniques。

Abstract：Background Prostatodynia remains a difficult clinical problem. Resiniferatoxin (RTX), an ultrapotent vanilloid, can produce a selective and long-lasting desensitization of nociception via C-fiber sensory neurons. Substance P (SP) and calcitonin gene-related peptide (CGRP) released from C-fibers are key neurotransmitters in visceral pain. In this study,we evaluated the analgesic effect of intrathecal RTX on rat prostatodynia.Methods Male Sprague-Dawley rats were divided into 3 groups for different treatment. In group A, sham operation was preformed. In group B, 100 μl complete Freund's adjuvant (CFA) was injected into the rat's bilateral ventral prostate to induce chronic inflammation. In group C, after prostatitis formed, 50 μl 10 nmol/L RTX was injected into the rat's lumbosacral (L5-S2) vertebral canal. SP and CGRP contents in the spinal cord were investigated by immunohistochemistry and radioimmunoassay (RIA). Their transcriptional levels in dorsal root ganglion (DRG) were determined by reverse transcriptase polymerase chain reaction (RT-PCR). In addition, pelvic nerve afferent discharge was recorded to explore the neuro-electrophysiological mechanisms underlying RTX-induced effect.Results SP and CGRP released in the spinal cord and their synthesis in DRG were increased significantly in response to CFA-induced chronic prostatitis, whereas this increase was effectively inhibited by intrathecal RTX. Meanwhile, pelvic nerve afferent electrical activity was enhanced significantly in rats with chronic prostatitis, but it was attenuated markedly in RTX-treated rats paralleled by the change of neuropeptides.Conclusions Intrathecal RTX administration could produce an analgesic effect on rat prostatodynia. Suppression of pelvic nerve afferent electrical activity may be a crucial mechanism underlying RTX-induced analgesia. RTX intrathecal application may present a novel analgesic strategy of prostatodynia。

Abstract：Background Recent studies have suggested that estrogens are involved in normal and abnormal prostate growth,though their exact role is still controversial. Oestrogens exert inhibitory and stimulatory effects on prostate gland, but the expression of oestrogen receptor-α (ERα) and oestrogen receptor-β (ERβ) in malignant prostate tissue remains unresolved. We determined ERα and ERβ in prostate cancer and investigated the relationship between expression of ER and pathological features of prostate carcinoma.Methods Thirty-two cases of prostate cancer, 12 cases of normal prostate tissue and 32 cases of benign prostate hyperplasia were analyzed for the expression of ERα and ERβ using semiquantitative, reverse transcription polymerase chain reaction (RT-PCR) and the products sequenced.Results Comparisons of the normal, hyperplastic and tumour prostate tissues indicated an overexpression of ERα in tumour specimens (P＜0.01). However, the expression of ERβ significantly reduced in tumour tissues compared with normal and hyperplastic specimens (P＜0.01), suggesting that severe pathological features of prostate cancer were associated with lower ERβ expression. Spearman analysis showed negative correlation between ERβ expression and tumour stage, grade (-0.67, -0.43, respectively, both P＜0.05), and a positive correlation between ERα expression and tumour stage, grade (0.51, 0.57, respectively, both P＜0.01). Our analysis also showed that hormone refractory, prostate cancer, compared with hormone dependent, prostate cancer, displayed a decreased expression of ERβ (P＜0.01) and an increased expression of ERα.Conclusions ERa and ERβ may play important roles in the development of prostate cancer. The decrease in ERβ expression is associated with higher Gleason grade tumours and prostate cancer with higher metastatic potential. The loss of ERβ could be one of the key processes leading to uncontrolled growth of prostate epithelial cells。

Abstract：Background E-cadherin, beta-catenin, cathepsin D, matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 are all invasion-related proteins. The expression patterns of these proteins in invasive ductal breast carcinomas, and their associations with known clinicopathological parameters, tumor recurrence and expressions of estrogen receptor (ER), progesterone receptor (PR), PS2 and c-erbB2 were not well studied in Chinese patients.Methods In a set of 94 invasive ductal breast carcinomas, protein expressions of these molecular markers were investigated by immunohistochemistry, and their associations with known clinicopathological parameters, tumor recurrence and expressions of ER, PR, PS2 and c-erbB2 were also examined. In addition, the interrelationship between the expressions of these proteins were studied.Results Preserved membrane E-cadherin expression was associated with late tumor stage and tumor recurrence,whereas the reduced junctional beta-catenin associated with positive lymph node status and c-erbB2 overexpression.Positive staining of cathepsin D in tumor stromal cells displayed a significant association with late tumor stage. High expression of MMP-2 in cancer cells was associated with large tumor size and PR positive expression. TIMP-2 expression was positively associated with tumor recurrence. In addition, inter-relationship between the expressions of these biomarkers was also assessed. Cathepsin D staining in cancer cells was inversely correlated with its staining in stromal cells, and also inversely correlated with MMP-2 staining in tumor stromal cells. MMP-2 expression in stromal cells displayed an inverse correlation with TIMP-2 expression. MMP-9 expression displayed parallel associations with TIMP-1 and TIMP-2 expression.Conclusion Evaluation of E-cadherin, beta-catenin, cathepsin D, MMP-2 and TIMP-2 expression may be of some help in more accurately predicting the prognosis of invasive ductal breast carcinomas。

Abstract：Background Many researches suggested that obesity increased the risk of breast cancer, but the mechanism was currently unknown. Adipocytokines might mediate the relationship. Our study was aimed to investigate the relationship between serum levels of resistin, adiponectin and leptin and the onset, invasion and metastasis of breast cancer.Methods Blood samples were collected from 80 newly diagnosed, histologically confirmed breast cancer patients and 50 age-matched healthy controls. Serum levels of resistin, adiponectin and leptin were determined by enzyme-linked immunosorbent assays (ELISA); fasting blood glucose (FBG), lipids, body mass index (BMI), and waist circumference (WC) were assayed simultaneously.Results Serum levels of adiponectin ((8.60±2.92) mg/L vs (10.37±2.81) mg/L, P=0.001) and HDL-c were significantly decreased in breast cancer patients in comparison to controls. Serum levels of resistin ((26.35±5.36) μg/L vs (23.32±4.75)μg/L, P=0.000), leptin ((1.35±0.42) μg/L vs (1.06±0.39) μg/L, P=0.003), FBG and triglyceride (TG) in breast cancer patients were increased in contrast to controls, respectively. However, we did not find the significant difference of the serum levels of resistin, adiponectin and leptin between premenopausal breast cancer patients and healthy controls (P=0.091, 0.109 and 0.084, respectively). The serum levels of resistin, adiponectin and leptin were significantly different between patients with lymph node metastasis (LNM) and those without LNM (P=0.001, 0.000 and 0.006, respectively).The stepwise regression analysis indicated that the tumor size had the close correlation with leptin (R2=0.414, P=0.000)and FBG (R2=0.602, P=0.000). Logistic regression analysis showed that reduced serum levels of adiponectin (OR:0.805;95%CI: 0.704-0.921; P=0.001), HDL (OR: 0.087; 95%CI: 0.011-0.691, P=0.021), elevated leptin (OR:2.235;95%CI:1.898-4.526; P=0.004) and resistin (OR: 1.335; 95%CI: 1.114-2.354; P=0.012) increased the risk for breast cancer; Reduced serum levels of adiponectin (OR: 0.742; 95%CI: 0.504-0.921; P=0.003) and elevated leptin (OR: 2.134;95%CI:1.725-3.921; P= 0.001) were associated with lymph node metastasis of breast cancer.Conclusions The decreased serum adiponectin levels and increased serum resistin and leptin levels are risk factors of breast cancer. The low serum adiponectin levels and high serum leptin levels are independent risk factors for metastasis of cancer. The association between obesity and breast cancer risk might be explained by adipocytokines。

Abstract：Background Definitive treatment for heart attack is early reperfusion with either angioplasty or thrombolytic therapy,and the benefit is strictly time-dependent. Patient outcomes are improved with either therapy when initiated as soon as possible. Recognition of heart attack symptoms is logically tied to taking action to receive prompt emergency care.Inadequate knowledge of heart attack symptoms may prolong delay. The purpose of this study was to document knowledge about heart attack symptoms in Beijing residents and to identify the characteristics associated with increased knowledge of heart attack.Methods A structured survey was conducted in 18 communities in Beijing from March 1 through June 10 in 2006.Addresses and participants were selected randomly following a stratification. The survey was designed to collect knowledge of heart attack symptoms from sampled adults in each community.Results A total of 4627 respondents completed the questionnaires correctly, and 50.29% of them were female. Totally 64.15% of the respondents reported chest pain or discomfort (common symptoms) as a symptom of heart attack; 75.38% reported at least one of the following eight symptoms as a symptom of heart attack: back pain, shortness of breath, arm pain or numbness, nausea or vomiting, neck, jaw or shoulder pain, epigastric pain, sweating, weakness (less common symptoms); 20.36% correctly reported four or more heart attack symptoms, only 7.4% knew all the correct heart attack symptoms, and 28.94% knew about reperfusion therapy for heart attack; 31.7% reported to call 120 or 999 while having a heart attack themselves; however 89.6% reported to call 120 or 999 when someone else is suffering from a heart attack. Very old persons and those with health insurance coverage, high education level, high household income, longer living in Beijing and previous experience with heart disease had greater knowledge of heart attack symptoms.Conclusions Public knowledge of common heart attack symptoms as well as less common heart attack symptoms is deficient in Beijing residents. But their knowledge of calling emergency medical services when someone is having a heart attack is relatively adequate. Public health efforts are needed to increase the recognition of the major heart attack symptoms in both the general public and groups at high risk for an acute cardiac event, especially in socioeconomically disadvantaged subgroups, including persons with low education level, low household income, and no health insurance coverage。

Abstract：Background The peripheral enhancement of small hepatocellular carcinoma (SHCC) is a rare appearance in dual phase images by helical computed tomography (CT). This study discusses this phenomenon and its correlative histopathology.Methods The helical CT dual phase appearance of peripheral enhancement in SHCC was analyzed in 21 cases (22 lesions). All lesions were confirmed as SHCC by histopathological examination.Results In these 22 lesions, enhanced peripheral ring in 20 lesions was incomplete, the thickness of enhanced peripheral ring varied and mural node could be found in hepatic arterial phase; only 2 lesions had complete peripheral ring enhancement and ring of uniform thickness in hepatic arterial phase. The enhancement of some peripheral rings and mural nodes dropped to very low density in portal venous phase. The tumour cells were grade Ⅰ in 3 lesions, Ⅱ in 16, Ⅲ in 2 and Ⅳ in 1. The vascular supply was more abundant at the border than in the centre of 15 lesions and the vascular supply was deficient in both centre and border of the remaining 7 lesions. In 3 lesions, the pseudocapsule showed in the border of the lesion. In 12 lesions, flecks of necrosis were found in the border and/or centre of the lesion.Conclusions The characteristic peripheral enhancement in helical CT dual phase images of small hepatocellular carcinoma correlates with different vascular supplies, fibrous capsule and necrosis of the lesion。

Abstract：Background It is known that the hypothalamic-pituitary-adrenal (HPA) axis is highlighted by stimulation, such as sepsis,trauma, etc, when corticortropin increases and plasma cortisol levels enhance. Relative adrenal insufficiency is not uncommon in critically ill patients and may occur in severe sepsis patients with high plasma cortisol levels. It has been demonstrated that a short corticotropin test has a good prognostic value and is helpful in identifying patients with septic shock at high risk for death, but it has not been established for all severe sepsis patients, especially in China. The aim of this study is to explore the relationship between adrenal function and prognosis in patients with severe sepsis.Methods This prospective study was conducted between July and December 2004 in 6 teaching hospitals. Two hundred and forty patients with severe sepsis were enrolled in this study. A short corticotropin stimulation test was performed in all patients by intravenous injection of 250 μg of corticotropin. Blood samples were taken immediately before the test (T0), 30 (T30) and 60 (T60) minutes afterward, and the plasma cortisol concentration was measured by radio-immunoassay. At the onset of severe sepsis, the following parameters were recorded: age, sex, Acute Physiology and Chronic Health Evaluation (APACHE) Ⅱ, heart rate, mean arterial pressure (MAP), arterial partial pressure of oxygen (PaO2)/fraction of inspired oxygen (FiO2), peripheral blood of hemoglobin, platelets and leukocyte concentration and the number of organ failure. Patients were designated into two groups (survival and non-survival groups) according to the 28-day mortality. Relative adrenal insufficiency was defined as the difference between T0 and the highest value of T30 or T60 (△Tmax) ≤9 μg/dl.Results (1) Two hundred and forty patients with severe sepsis were included in this study, with 134 patients in the survival group and 106 in the non-survival group. The 28-day mortality was 44.2%. (2) Between the survival group and non-survival group age, APACHE Ⅱ, peripheral blood of platelets, the number of organ failures, T0 and △Tmax showed significant differences. T0 was (23±10) μg/dl and (36±18) μg/dl in the survival group and nonsurvival group respectively.△Tmax was (18±9) and (10±8) μg/dl in the survival group and non-survival group respectively. The areas under the ROC curve for T0 and △Tmax were both 0.72, and the area under the ROC curve for APACHE Ⅱ was 0.70. By multivariate analysis age, T0, the number of organ failures and relative adrenal insufficiency (△Tmax ≤9 μg/dl) were independent predictors of death. (3) The incidence of relative adrenal insufficiency was 38.3% in total, 19.4% in the survival group and 62.3% in the non-survival group (P＜0.001). The 28-day mortality was 71.7% among the relative adrenal insufficiency patients but 27.0% among normal adrenal function patients.Conclusions The prevalence of relative adrenal insufficiency is high in severe sepsis. Relative adrenal insufficiency has a good prognostic value for severe sepsis。

Abstract：Background Compensatory sweating (CS) is one of the most common postoperative complications after thoracic sympathectomy, sympathicotomy or endoscopic sympathetic block (ESB) for palmar hyperhidrosis. This study was conducted to examine the relevance between CS and the sympathetic segment being transected in the surgical treatment of palmar hyperhidrosis, and thus to detect the potential mechanism of the occurrence of CS.Methods Between October 2004 and June 2006, 163 patients with primary hyperhidrosis were randomly divided into two groups, T3 sympathicotomy (78 patients) and T4 sympathicotomy(85), who were operated upon under general anesthesia via single lumen intubation and intercostal video-mediastinoscopy (VM).Results No morbidity or mortality occurred. Palmar hyperhidrosis was cured in all patients. Follow-up(mean (13.8±6.2)months) showed no recurrence of palmar hyperhidrosis. The difference of rates of mild CS in groups T3 and T4 was of no statistical significance. The rate of moderate CS was significantly lower in group T4 than in group T3. No severe CS occurred.Conclusion The rates of occurrence and severity of CS are lowered with the lower sympathetic chain being transected。

Abstract：Background Although the use of hepatitis B immunoglobulin (HBIG) may lead to a significant reduction in recurrent hepatitis B virus (HBV) infection and improve the survival of patients who have undergone liver transplantation (LT) for hepatitis B-related diseases, the recurrence of the disease still remains at a lower level. Different clinical curative effects were observed in patients with the same HBV-related diseases and the same therapy. This study was undertaken to investigate whether the efficacy of HBIG is associated with FCGR3A gene polymorphisms in Chinese liver transplant patients.Methods Altogether 77 patients who had received liver transplantation for hepatitis B-related diseases with more than one-year survival after surgery were studied. The recurrence of HBV was characterized by the appearance of HBsAg in serum after the operation. The FCGR3A genotyping was performed using genomic DNA sequencing (ABI 3037). Single nucleotide polymorphism at nucleotide 559 was detected by Polyphred.Results Of the 77 patients, 14 were complicated with HBV recurrence post-transplant. The FCGR3A at nucleotide 559 TT was observed in 35 (45.5%) subjects, whereas TG in 31(40.3%) and GG in 11(14.3%). In the 559G carrier group (n=42, 54.5%), the risk of HBV recurrence was 9.5%, and 1- and 2-year recurrence-free survival rates were 95.2% and 88.7%, respectively. In the 559G noncarrier group (n=35, 45.5%), the risk of HBV recurrence was 28.6%, and 1- and 2-year recurrence-free survival rates were 74.3% and 69.3%, respectively. The risk of HBV recurrence and the recurrence-free survival rate were both statistically different between the 559G carrier and noncarrier groups (P＜0.05).Conclusions A single nucleotide polymorphism (T/G) at position 559 of the FCGR3A gene was found in Chinese patients. The efficacy of HBIG in prophylaxis of HBV recurrence after LT is associated with the gene polymorphism, so detecting FCGR3A genotypes can be a clinical reference of the HBIG administration。

Protein kinases C (PKCs), a big family including 12 isoenzymes, are divided into three major groups according to the variability of their regulatory domains。

HIV-1 Nef protein plays an important role in HIV-1 pathogenesis, including downregulation of the cell surface CD4 and class Ⅰ major histocompatibility complex (MHC), enhancement of virion infectivity and alteration of a variety of cellular signal transduction pathways。

Epidemiologic investigations have shown that the morbidity of obstructive sleep apnea syndrome (OSAS) among adults is 2%-4%, among the population aged 30 years and over is 4.63%, and among patients with hypertension or coronary atherosclerostic disease (CAD) is as high as 30%-50%。

Adult idiopathic thrombocytopenic purpura (ITP) is a chronic acquired organ-specific autoimmune hemorrhagic disease characterized by the production of auto-antibodies against antigens on the membranes of platelet, resulting in enhanced Fc-mediated destruction of the platelets by macrophages in the reticuloendothelial system。

To the Editor: With interest we read the article by Wu et al1 on a 53-year male with agenesia of the right lung and left-ventricular hypertrabeculation / noncompaction (LVHT). The paper raises the following concerns。

To the Editor: We read with great interest the letter from Drs. Finsterer and St(o)llberger, on our case report.1 We are grateful to their comments。