生殖器流行性卡波西氏肉瘤71例临床分析

目的 :分析生殖器流行性卡波西氏肉瘤 (Kaposi sSarcoma简称KS)的发病情况、临床特征及治疗方法。资料和方法 :1986年 6月至 1996年 12月 ,巴黎第十二大学医学院HenriMondor医院肿瘤放疗科共收治 64 5例爱滋病KS病人 ,其中 71例有阴茎及阴囊病变。单发于阴茎或阴囊有 12例 ,其余 5 9例均合并有其它部位的皮肤侵犯。中位年龄 3 7岁 ,9例有排尿困难 ,13例有水肿或 /和疼痛。均采用放射治疗 ,选用低能 ( 4 5KV)X线 ,大野照射10Gy/4次·周 ,根据病变反应 ,休息 15天 ,缩野再予 5～ 10Gy。结果 :所有病人均获客观上的缓解和器官功能的恢复。在 71例共 189个病变部位中 ,完全反应率 2 8% ,部分反应率 66% ,6%无反应。所有病人均获随访 ,平均随访期 7 5个月。结论 :发生于生殖器的流行性KS少见。放疗对本病可取得满意结果 ;

Effects of ursolic acid and oleanolic acid on human colon carcinoma cell line HCT15

AIM: Ursolic acid (UA) and oleanolic acid (OA) aretriperpene acids having a similar chemical structure and aredistributed wildly in plants all over the world. In recentyearn, it was found that they had marked anti-tumor effects.There is little literature currently available regarding theireffects on colon carcinoma cells. The present study wasdesigned to investigate their inhibitory effects on humancolon carcinoma cell line HCT15 METHODS: HCT15 cells were cultured with different drugs.The treated cells were stained with hematoxylin-eosin andtheir morphologic changes observed under a lightmicroscope. The cytotoxicity of these drugs was evaluatedby tetrazolium dye assay. Cell cycle analysis was performedby flow cytometry (FCM). Data were expressed as means +SEM and Analysis of variance and Student' t-test forindividual comparisons.RESULTS: Twenty-four to 72 h after UA or OA 60 μmol/Ltreatment, the numbers of dead cells and cell fragmentswere increased and most cells were dead at the 72 nd hour.The cytotoxicity of UA was stronger than that of OA.Seventy-eight hours after 30 μmol/L of UA or OA treatment,a number of cells were degenerated, but cell fragments wererarely seen. The IC50 values for UA and OA were 30 and 60μmol/L, respectively. Proliferation assay showed thatproliferation of UA and OA-treated cells was slightlyincreased at 24 h and significantly decreased at 48 h and 60h, whereas untreated control cells maintained anexponential growth curve. Cell cycle analysis by FCMshowed HCT15 cells treated with UA 30 and OA 60 for 36 h and72 h gradually accumulated in G0/G1 phase (both drugs P＜ 0.05 for 72 h), with a concomitant decrease of cell populationsin S phase (both drugs P＜ 0.01 for 72 h) and no detectableapoptotic fraction.CONCLUSION: UA and OA have significant anti-ttumor activity.The effect of UA is stronger than that of OA. The possiblemechanism of action is that both drugs have an inhibitoryeffect on tumor cell proliferation through cell-cycle arrest.

Expression of fragile histidine triad in primary hepatocellular carcinoma and its relation with cell proliferation and apoptosis

AIM: To evaluate the expression of fragile histidine triad (FHIT) gene protein, product of a candidate tumor suppressor,and to investigate the relationship between FHIT, cell apoptosis and proliferation, and pathological features of primary hepatocellular carcinoma (HCC).METHODS: Forty-seven HCC and ten normal liver specimens were collected during surgical operation between 2001and 2003. FHIT and proliferating cell nuclear antigen (PCNA)expression were detected by immunohistochemistry, and apoptotic level was evaluated by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay on the tissue sections.RESULTS: All normal liver tissues showed a strong expression of FHIT, whereas 28 of 47 (59.6%) carcinomas showed a significant loss or absence of FHIT expression (P = 0.001).The proportion of reduced FHIT expression in those carcinomas at stages Ⅲ-Ⅳ (70.6%) and in those with extrahepatic metastasis (86.7%) showed an increasing trend compared with those at stages Ⅰ-Ⅱ (30.8%, P= 0.013) and those without metastasis (46.9%, P = 0.010) respectively. Apoptotic incidence in advanced TNM stage carcinoma and those with positive FHIT expression was higher than that in early stage carcinoma (P = 0.030) and in those with negative FHIT expression (P = 0.044) respectively. The proliferating potential of hepatocellular carcinoma was associated with FHIT expression (P = 0.016) and the aggressive feature (P = 0.019). Kaplan-Meier analysis demonstrated that the survival time of these 47 patients correlated with TNM stage,FHIT expression and metastasis.CONCLUSION: There is marked loss or absence of FHIT expression, as well as abnormal apoptosis-proliferation balance in HCC. FHIT may play an important role in carcinogenesis and development of HCC.

Expression of vascular endothelial growth factor-C and the relationship between lymphangiogenesis and lymphatic metastasis in colorectal cancer

AIM: To investigate the expression of vascular endothelial growth factor-C (VEGF-C) and the relationship between VEGF-C and lymphangiogenesis, lymph node metastasis in colorectal cancer. METHODS: Fifty six cases of colorectal cancer were selected randomly. Expression of VEGF-C was detected by immunohistochemistry, and lymphatic vessels were stained by enzyme histochemical method. RESULTS: VEGF-C expression was found in 66.7% (37/56) patients. In VEGF-C positive and negative patients, the lymphatic vessel density was 25.16±7.52 and 17.14±7.22, respectively (P<0.05). The rate of lymph node metastasis in VEGF-C positive patients (81.1%) was significantly higher than that in the negative group (42.1%). CONCLUSION: VEGF-C expression may induce lymphangiogenesis in colorectal cancer, as a result, tumor cells can entry the lymphatic vessels easily. VEGF-C may serve as a useful prognotic factor in colorectal carcinoma.

Hemizygous deletion and hypermethylation of RUNX3 gene in hepatocellular carcinoma

AIM：To analyze the genetic and epigenetic alterations of RUNX3 gene, a potential putative tumor suppressor gene,in hepatocellular carcinoma (HCC).METHODS: PCR-based loss of heterozygosity (LOH) detection, analysis of mutation with PCR-single strand conformational polymorphism (SSCP) and sequencing, and methylation study with methylation specific PCR (MSP) were performed on RUNX3 gene in a series of 62 HCCs along with their matched normal tissues.RESULTS:Mutation of RUNX3 gene was not found, but one single nucleotide polymorphism with T to A transversion at the second nucleotide of the 18th condon was found.Nine of 26 informative cases (34.6%) showed allelic loss on the polymorphic site and 30 cases (48.4%) revealed hypermethylation of RUNX3 gene in promoter CpG islands.Furthermore,of the 9 cases with LOH, 8 (88.9%) also had hypermethylation.CONCLUSION:Our findings indicate that inactivation of RUNX3 gene through allelic loss and promoter hypermethylation might be one of the major mechanisms in hepatocellualr carcinogenesis.

Comparison between chemoembolization combined with radiotherapy and chemoembolization alone for large hepatocellular carcinoma

AIM: To investigate the efficacy of transcatheter arterial chemoembolization (TACE) combined with radiotherapy for unresectable large hepatocellular carcinoma (HCC).METHODS: From June 1994 to June 1999, a total of 76patients with large unresectable HCC were treated with TACE followed by external-beam irradiation. 89 patients with large HCC, who underwent TACE alone during the same period,served as the control group. Clinical features, therapeutic modalities, acute effects and survival rates were analyzed and compared between TACE plus irradiation group and TACE alone group. A multivariate analysis of nine clinical variables and one treatment variable (irradiation) was performed by the Cox proportional hazards model.RESULTS: The clinical features and therapeutic modalities except irradiation between the two groups were comparable (P＞0.05). The objective response rate (RR) in TACE plus irradiation group was higher than that in TACE alone group (47.4 % vs28.1%, P＜0.05). The overall survival rates in TACE plus irradiation group (64.0 %, 28.6 %, and 19.3 %at 1, 3, 5 years, respectively) were significantly higher than those in TACE alone group (39.9 %, 9.5 %, and 7.2%,respectively, P=0.0001). Cox proportional hazards model analysis showed that tumor extension and Child grade were significant and were independent negative predictors of survival, while irradiation was an independent positive predictor of survival.CONCLUSION: TACE combined with radiotherapy is more effective than TACE alone, and is a promising treatment for unresectable large HCC.

Expression of TIMP-1 and TIMP-2 in rats with hepatic fibrosis

AIM: To investigate the location and expression of TIMP-1 and TIMP-2 in the liver of normal and experimental hepatic fibrosis in rats. METHODS: The rat models of experimental immunity hepatic fibrosis (n=20) were prepared by the means of immunologic attacking with human serum albumin (HSA),and normal rats (n=10) served as control group. Both immunohistochemistry and in situ hybridization methods were respectively used to detect the TIMP-1 and TIMP-2 mRNA and related antigens in liver. The liver tissue was detected to find out the gene expression of TIMP-1 and TIMP-2 with RT-PCR. RESULTS: The TIMP-1 and TIMP-2 related antigens in livers of experimental group were expressed in myofibroblasts and fibroblasts (TIMP-1: 482±65 vs 60±20; TIMP-2:336±48 vs 50±19, P<0.001). This was the most obvious in portal area and fibrous septum. The positive signals were located in cytoplasm, not in nucleus. Such distribution and location were confirmed bysitu hybridization (TIMP-1/β-actin: 1.86±0.47 vs 0.36±0.08; TIMP-2/β-actin: 1.06±0.22 vs 0.36±0.08,P<0.001). The expression of TIMP-1 and TIMP-2 was seen in the liver of normal rats, but the expression level was very low. However, the expression of TIMP-1 and TIMP-2 in the liver of experimental group was obviously high. CONCLUSION: In the process of hepatic fibrosis, fibroblasts and myofibroblasts are the major cells that express TIMPs.The more serious the hepatic fibrosis is in the injured liver,the higher the level of TIMP-1 and TIMP-2 gene expression.

Expression of p57^(kip2) and its relationship with clinicopathology, PCNA and p53 in primary hepatocellular carcinoma

AIM: To investigate the expression of p57kip2 and its relationship with dinicopathology, PCNA and p53 in primary hepatocellular carcinoma (HCC).METHODS: Expression of p57kip2, PCNA and p53 in tumor tissues from 32 patients with HCC and 10 liver tissues of normal persons was detected with Elivision immunohistochemical technique.RESULTS: The p57kip2 protein positive-expression rate in HCC was 56.25%, lower than that in normal tissues (100%, P＜0.05). The reduced expression of p57kip2 protein correlated significantly with moderate or low differentiation of tumor cells (P = 0.007 ＜0.05), high clinical stage (P = 0.041 ＜0.05) and poor prognosis (P = 0.036 ＜0.05),but did not correlate significantly with metastasis, tumor size, level of AFP and age (P＞0.05). The PCNA positiveexpression rate was 56.25%, which was correlated significantly with the expression of p57kip2 (P= 0.025＜0.05).The p53 positive-expression rate was 46.88%, which was not correlated significantly with the expression of p57kip2(P＞0.05).CONCLUSION: There is a marked loss or absence of p57kip2 expression and high expression of PCNA in HCC,which are involved in carcinogenesis and development of HCC. The p57kip2 and p53 may induce apoptosis via different mechanisms.

Tumor suppressor gene p16 and Rb expression in gastric cardia precancerouslesions from subjects at a high incidence area in northern China

AIM: To further understand the molecular basis for gastriccardia carcinogenesis and to provide etiological clues.METHODS: Endoscopic mucosa biopsy and histopathologicalexaminations were made on 37 subjects from a high incidencearea for both esophageal and gastric cardia carcinomas innorthem China. All the biopsy samples were fixed in 850 mi. -1 Lalcohol and embedded in paraffin. Each block contained onepiece of tissue and was serially section at 5 μm.Immunohistochemistry (ABC) was carried out on these gastriccardia samples to determine the alterations of p16 and Rb.RESULTS: Based on the histopathlogical examinationtherewere 11 cases of chronic superficial gastritis, 12 cases ofchronic atrophic gastritis and 14 cases of dysplasia. Theimmunostaining demonstrated different levels of unclearimmunostaining of p16 and Rb in normal gastric cardiatissue and the tissues with different severity of lesions. Withthe lesions progressing, the positive immunostaining ratesfor pi6 protein had a decreasing tendency. In contrast, thepositive immunostaining rate for Rb protein had anincreasing tendency. There was a significant negativerelationship between the two parameters. Changes of p16wasCSG 11(100 % ), CAG 7(58 % ), DYS 4(29 % ) andchanges of Rb was CSG 2(18 %), CAG 8(67 %) and DYS 12(86 %), (p＜0.05).CONCLUSION: The alterations of p16 and Rb protein may playa role in the early stages of gastric cardia carcinogenesis.

Effect of intraoperative radiotherapy combined with external beam radiotherapy following internal drainage for advanced pancreatic carcinoma

AIM: To determine the survival of advanced pancreatic cancer patients treated with intraoperative radiotherapy (IORT) combined with external beam radiation therapy (EBRT) following internal drainage (cholecystojejunostomy or choledochojejunostomy). METHODS: Eighty-one patients with advanced pancreatic cancer who received IORT combined with EBRT following internal drainage (ID) between 1996 and 2001 were retrospectively analyzed. Among the 81 patients, 18 underwent ID+IORT, 25 ID+IORT+EBRT (meanwhile, given 5-Fu 300 mg/m^2 iv drip, 2f/w), 16 EBRT, 22 had undergone simple internal drainage. The IORT dose was 15-25Gy in a single fraction. The usual EBRT dose was 30-40Gy with a daily fraction of 1.8-2.0 Gy. RESULTS: The complete remission rate, partial remission rate of patients with backache and abdominal pain treated with ID+IORT were 55.5%, 33.3% respectively. Alleviation of pain was observed 2 or 3 wk after IORT. The median survival time (MST) of ID+IORT group was 10.7 mo. The pain remission rate of patients treated with ID+IORT+EBRT was 92%, and their MST was 12.2 mo. The MST of patients treated with EBRT and simple internal drainage was 5.1 mo and 7.0 mo, respectively. The survival curve of ID+IORT group and ID+IORT+EBRT group was significantly better than that of EBRT group (P<0.05). The difference between the ID+IORT+EBRT group and ID group was significant (P<0.05). CONCLUSION: IORT combined with EBRT following internal drainage can alleviate pain, improve quality of life and prolong survival time of patients with advanced pancreatic cancer.

Surgical treatment and prognosis of gastric cancer in 2613 patients

AIM: To analyze the factors influencing the prognosis of patients with gastric cancer after surgical treatment, in order to optimize the surgical procedures.METHODS: A retrospective study of 2 613 consecutive patients with gastric cancer was performed. Of these patients, 2 301 (88.1%) received operations; 196 explorative laparotorny (EL), 130 by-pass procedure (BPP), and 1 975 surgical resection of the tumors (891 palliative resection and 1 084 curative resection). The survival rate was calculated by theactuarial life table method, and the prognostic factors were evaluated using the Cox regression proportional hazard model.RESULTS: Of the patients, 2 450 (93.8%) were followed-up.The median survival period was 4.6 mo for patients without operation, 5.2 mo for EL, 6.4 mo for BPP, and 15.2 mo for palliative resection (P = 0.0001). Of the patients with surgical resection of the tumors, the overall 1, 3 and 5-yearsurvival rates after were 82.7%, 46.3% and 31.1%,respectively, with the 5-year survival rate being 51.2% in patients with curative resection, and 7.8% for those with palliative resection. The 5-year survival rate was 32.5% for patients with total gastrectorny, and 28.3% for those with total gastrectomy plus resection of the adjacent organs. The factors that independently correlated with poor survival included advanced stage, upper third location, palliative resection, poor differentiation, type IV of Borrman nclassification, tumor metastasis (N3), tumor invasion into the serosa and contiguous structure, proximal subtotal gastrectomy for upper third carcinoma and D1 lymphadenectomy aftercurative treatment.CONCLUSION: The primary lesion should be resected as long as the local condition permitted for stage III and IV tumors, in order to prolong the patients' survival and improve their quality of life after operation. Total gastrectomy is indicated for carcinomas in the cardia and fundus, and gastric cancer involving the adjacent organs without distant metastasis requires gastrectomy with resection of the involved organs.

Influence of survivin and caspase-3 on cell apoptosis and prognosis in gastric carcinoma

AIM: To evaluate the role of survivin and caspase-3 in apoptosis of gastric carcinoma, as well as in prognosis of patients with gastric carcinoma.METHODS: Expressions of survivin and caspase-3 were investigated immunohistochemically in 80 gastric carcinoma patients without a history of chemo-radiation therapy. Tumor cell apoptosis was examined by TUNEL method.RESULTS: Immunohistochemical analysis showed that survivin expression was positive in 61 of 80 patients (76%) with gastric carcinoma. In contrast, no expression of survivin in adjacent normal tissues was detected. Expression level of caspase-3 was higher in normal tissues than in carcinoma.Patients with higher expression of survivin had worse histological grades and pathological stages. Expression of caspase-3 was significantly associated with histological stages, but not with the pathological stages. Although survivin expression in carcinoma was not inversely related to caspase-3, patients with survivin (-) and caspase-3(+) had the maximum apoptosis index.CONCLUSION: Expression level of survivin was associated with histological grades and pathological stages of the tumor,indicating that survivin may be a poor prognosis factor for gastric carcinoma. Unlike caspase-3, survivin (an apoptosis inhibitor) can markedly inhibit the apoptosis of tumor cells.

Expression of cyclooxygenase-2 in colorectal cancer and its clinical significance

AIM: To clarify the clinicopathologic significance of COX-2expression in human colorectal cancer.METHODS: A total of 128 surgically resected colorectal cancer specimens were immunohistochemically analyzed with the use of anti-COX-2, anti-VEGF and antiMMP-2 antibodies. The relationship between the cyclooxygenase-2 expression in primary lesions of colorectal cancer and clinicopathologic parameters was evaluated by chi-square test.RESULTS: Among 128 cases of colorectal cancer, 87(67.9%) were positive for cyclooxygenase-2. The expression of cyclooxygenase-2 was significantly correlated with the depth of invasion, stage of disease,and metastasis (lymph node and liver). Patients in T3-T4,stages Ⅲ-Ⅳ and with metastasis had much higher expression of cyclooxygenase-2 than ones in T1-T2, stages Ⅰ-Ⅱ and without metastasis (P＜0.05). Among 45 cases of colorectal cancer with lymph node metastasis, the COX-2-positive rate was 86.7% (39/45) for primary lesions and diffuse cytoplasmic staining for COX-2 protein was detected in cancer cells in 100% of metastatic lesions of the lymph nodes. VEGF expression was detected in 49tumors (38.3%), and VEGF expression was closely correlated with COX-2 expression. The positive expression rate of VEGF (81.6%) in the cyclooxygenase-2-positive group was higher than that in the cyclooxygenase-2-negative group (18.4%, P＜0.05). MMP-2 expression was detected in 88 tumors (68.8%), and MMP-2 expression was closely correlated with COX-2 expression. The positive expression rate of MMP-2 (79.6%) in the positive COX-2group was higher than that in the negative COX-2 group (20.4%, P＜0.05).CONCLUSION: Cyclooxygenase-2 may be associated with tumor progression by modulating the angiogenesis and cancer cell motility and invasive potential in colorectal cancer and it can be used as a possible biomarker.

Clinicopathological significance of expression of paxillin, syndecan-1 and EMMPRIN in hepatocellular carcinoma

AIM: To evaluate the relationship of expression of paxillin,syndecan-1 and EMMPRIN proteins with clinicopathological features in hepatocellular carcinoma (HCC).METHODS: Fifty-one patients who underwent HCC resection were recruited in the study. Paxillin, syndecan1 and EMMPRIN proteins in HCC tissues were detected with immunohistochemical staining.RESULTS: Of 51 cases of HCC, 23 (45%) exhibited paxillin protein positive expression. Of 42 cases of adjacent nontumor liver tissues, 24 (57%) exhibited positive expression.Positive paxillin protein expression was associated with low differentiation (r= 0.406, P= 0.004), with the presence of portal vein thrombosis (r = 0.325, P = 0.021), with extra-hepatic metastasis (r = 0.346, P = 0.014). Of 51cases of HCC, 28 (55%) exhibited syndecan-1 protein positive expression. Of 42 cases of adjacent non-tumor liver tissues, 23 (55%) exhibited positive expression.Positive snydecan-1 protein expression was associated with well differentiation (r = 0.491, P = 0.001), with no extra-hepatic metastasis (r = 0.346, P = 0.014). Of 51cases of HCC, 28 (55%) exhibited EMMPRIN protein positive expression. Of 42 cases of adjacent non-tumor liver tissues, 21 (50%) exhibited positive expression.Expression of EMMPRIN protein was not associated with serum AFP level, HBsAg status, presence of microsatellite nodule, tumor size, presence of cirrhosis and necrosis,differentiation, presence of portal vein thrombosis, extrahepatic metastasis, disease-free survival and overall survival (P＞0.05). Expression of paxillin protein was correlated conversely with the expression of syndecan-1protein in HCC (r = -0.366, P = 0.010).CONCLUSION: Expression of paxillin and syndecan-1proteins in HCC may affect its invasive and metastatic ability of the tumor. There may be a converse correlation between the expression of paxillin and syndecan-1 protein in HCC. Expression of EMMPRIN protein may be detected in HCC, but it may play little role in the invasion and metastasis of HCC.

Expression of Epstein-Barr virus genes in EBV-associated gastric carcinomas

AIM: To understand the expression of latent and lytic genes of Epstein-Barr virus (EBV) in EBV-associated gastric carcinoma (EBVaGC) and to explore the relationship between EBV-encoded genes and development of EBVaGC at molecular level.METHODS: One hundred and seventy-two gastric carcinoma tissues and 172 corresponding para-carcinoma tissues were tested for EBV genome by polymerase chain reaction (PCR)-Southern blotting. EBV-encoded small RNA (EBER)1 of the PCR positive specimens was detected by in situ hybridization (ISH). Gastric carcinomas with positive EBER1signals were classified as EBVaGCs. RT-PCR and Southern hybridization were applied to the detection of expression of nuclear antigen (EBNA) promoters (Qp, Wp and Cp),EBNA 1 and EBNA 2, latent membrane proteins (LMP) 1,2A and 2B and lytic genes (immediate early genes BZLF1and BRLF1, early genes BARF1 and BHRF1, late genes BcLF1and BLLF1) in EBVaGCs.RESULTS: Eleven EBV positive samples existed in gastric carcinoma tissues (6.39%). No EBV positive sample was found in corresponding para-carcinoma tissues. The difference between EBV positivity in carcinoma tissues and corresponding para-carcinoma tissues was significant(x2 = 9.0909, P = 0.0026). Transcripts of Qp and EBNA1were detected in all the 11 EBVaGCs, while both Wp and Cp were silent. EBNA2, LMP1 and LMP2B mRNA were absent in all the cases, while LMP2A mRNA was detected in 4 of the 11 cases. Of the 11 EBVaGCs, 7 exhibited BcLF1 transcripts and 2 exhibited BHRF1 transcripts. The transcripts of BZLF1and BARF1 were detected in 5 cases, respectively. No BLLF1and BRLF mRNA were detected.CONCLUSION: The latent pattern of EBV in gastric carcinoma corresponds to the latency Ⅰ/Ⅱ. Some lyric infection genes are expressed in EBVaGCs tissues. BARF1 and BHRF1genes may play an important role in tumorigenesis of gastric carcinoma.

Downregulation of retinoic acid receptor-β_z expression is linked to aberrant methylation in esophageal squamous cell carcinoma cell lines

AIM:To study the role of hypermethylation in the loss of retinoic acid receptor β2(RARβ2) in esophageal squamous cell carcinoma (ESCC).METHODS:The role of hypermethylation in RARβ2 gene silencing in 6 ESCC cell lines was determined by methylation-specific PCR (MSP),and its methylation status was compared with RARβ2 mRNA expression by RT-PCR.The MSP results were confirmed by bisulfite sequencing of RARβ2promoter regions.RESULTS:Methylation was detected in 4 of the 6 cell lines,and the expression of RARβ2 was markedly downregulated in 3 of the 4 methylated cell lines. The expression of RARβ2 was restored in one RARβ2-downregulated cell line with the partial demethylation of promoter region of RARβ2 after 5-aza-2′-deoxycytidine (5-aza-dc) treatment.CONCLUSION:The methylation of the 5′ region may play an important role in the downregulation of RARβ2 in some ESCC cell lines, suggesting that multiple mechanisms contribute to the loss of RARβ2expression in ESCC cell lines.This study may have clinical applications for treatment and prevention of ESCC.

Perioperative cimetidine administration promotes peripheral blood lymphocytes and tumor infiltrating lymphocytes in patients with gastrointestinal cancer:Results of a randomized controlled clinical trial

AIM: To study the effects of perioperative administration of cimetidine (CIM) on peripheral blood lymphocytes, natural killer (NK) cells and tumor infiltrating lymphocytes (TIL) in patients with gastrointestinal (GI) cancer. METHODS: Forty-nine GI cancer patients were randomized into treatment group, who took CIM in perioperative period,and control group, who did not take the drug. The treatment was initiated 7 days before operation and continued for 10 days after surgery. At baseline examination before operation, on the 2rid and 10th postoperative days, total T lymphocytes, T helper cells, T suppressor cells, and NK cells in peripheral blood were measured respectively by immunocytochemical method using mouse-anti human CD3,CD4, CD8 and CD57 monoclonal antibodies. Blood samples from 20 healthy volunteers were treated in the same way as normal controls. Surgical specimens were examined during routine histopathological evaluation for the presence of TiL in tumor margin. Immunohistochemical study was performed to measure the proportion of T and B lymphocytes in T[L population. T and B lymphocytes were detected respectively using mouse-anti-human CD3 and CD20 monoclonal antibodies. RESULTS: in comparison with normal controls, both the treatment and control groups had decreased T cells, T helper cells and NK cells at baseline. In control group, total T cells,T helper cells and NK cells declined continuously with the disease progression and the decrease became more obvious after operation. From baseline to the 2nd postoperative day,the proportion of total T cells, T helper cells, and NK cellswent down from 60.5±4.6% to 56.2±3.8%, 33.4±3.7% to28.1±3.4%, and 15.0±2.8% to 14.2±2.2%, respectively.On the other hand, there were significant improvements in these parameters after CIM treatment. On the 10th postoperative day, the treatment group had significantly higher percentages of total T cells, T helper cells and NK cells than control group. Moreover, CIM treatment also boosted TiL response, as was reflected by findings that 68%(17/25) of the patients in treatment group had significant TIL responses and only 25% (6/24) of the cases had discernible TIL responses (P<0.01). CONCLUSION: Perioperative application of CIM to GI cancer patients could help restore the diminished cellular immunity induced by tumor burden and surgical maneuver. The drug could also boost TIL responses to tumor. These effects suggest that the drug be used as an immunomodulator for GI cancer patients.

Effect of mutated IKBa transfection on multidrug resistance in hilar cholangiocarcinoma cell lines

AIM: To explore the expression effect of mutated IκBαtransfection on multidrug resistance gene (MDR-1) in hilar cholangiocarcinoma cells by inhibiting the activity of nuclear transcription factor-κB (NF-κB).METHODS: We used the mutated IκBα plasmid to transfect QBC939HCVC+ cells and QBC939 cells, and electrophoretic gel mobility shift assay (EMSA) to detect the binding activity of NF-κB DNA and the effect of the transfrecting mutated IκBα plasmid on multidrug resistance gene (MDR-1) in hilar cholangiocarcinoma cells and its expression protein (P-GP).RFSULTS: Plasmid DNA was digested by restriction enzymes Xbal and Hand Ⅲ, and its product after electrophoresis showed two bands with a big difference in molecular weight,with a size of 4.9 kb and 1.55 kb respectively, which indicated that the carrier was successfully constructed and digested with enzymes. The radioactivity accumulation of QBC939HCVC+and QBC939 cells transfected with mutated IκBα plasmid was significantly lower than that of the control group not transfected with mutated IκBα plasmid. Double densimeter scanning showed that the relative signal density between the tansfection group and non-transfection group was significantly different, which proved that the mutated IκBα plasmid could inhibit the binding activity of NF-κB DNA in hilar cholangiocarcinoma cells. Compared to control group not transfected with m IκBα plasmid, the expression level of MDR-1mRNA in the QBC939 and QBC939HCVC+ cells transfected with mutated IκBα plasmid was lower. The expression intensity of P-GP protein in QBC939 and QBC939HCVC+ cells transfected with mutated IκBα was significantly lower than that of the control group not transfected with mutated IκBα plasmid.CONCLUSION: The mutated IκBα plasmid transfection can markedly reverse the multidrug resistance of hilar cholangiocarcinoma cells. Interruption of NF-κB activity may become a new target in gene therapy for hilar cholangiocarcinogenesic carcinoma.

TIP30 regulates apoptosis-related genes in its apoptotic signal transduction pathway

AIM: To investigate the role of TIP30 in apoptotic signal pathway in hepatoblastoma cells and to provide a basis for TIP30 as a gene therapy candidate in the regression of hepatoblastoma cells.METHODS: Apoptosis of human hepatoblastoma cell lines HepG2 (p53 wild), Hep3B (p53 null) and PLC/RPF/5 (p53mutant) infected with Ad-TIP30 (bearing a wild type human Tip30 gene) were analyzed and p53, Bax and Bclxl expression levels were compared among these cells.MlT assay, DNA fragmentation, in situ 3' end labeling of DNA, annexin-Ⅴ FITC staining were used to detect cell death and apoptosis in cells at various time intervals subsequent to infection, and to determine whether TIP30 had an effect on the expression levels of some apoptosis-related gene products such as Bax, p53 and Bcl-xl. A similar time course experiment was performed by Western blotting.RESULTS: In MTT assay, the viability of HepG2 cells decreased significantly from 99.7% to 10% and displayed more massive cell death within 5-8 d than Hep3B and PLC/RPF/5 cells, with their viability decreased from 97.8% to 44.3% and 98.1% to 50.4%, respectively. In annexin-ⅤFITC assay, the percentage of apoptosis cells in HepG2cells was two to three-fold higher than that in control cells (infected with Ad-GFP), two-fold higher than that in Hep3B cells and 1.4-fold higher than that in PLC/RPF/5 cells 36 h after infection, respectively. Moreover, in HepG2 cells, the p53 began to increase 6-8 h after infection, reaching a maximum level between 8 and 12 h after infection and then dropped. Bax showed a similar increase in the cells as p53 reached the maximum at 8-12 h and subsequently decreased. Interestingly, Bcl-xl protein levels were down regulated during 24 to 36 h after Ad-TIP30 infection. In contrast, ectopic expression of TIP30 in Hep3B and PLC/RPF/5 cells had no effect on the regulation of Bax expression, but had an effect on Bcl-xl levels. In comparison with HepG2 cells, these data suggested that up-regulation of p53 levels by TIP30 might be a pre-requisite for Bax and Bax/Bcl-xl ratio increase. We hypothesized that TIP30 might regulate Bax gene partly through p53, which sensitizes cells to apoptosis by involving a p53 apoptosis signal transduction pathway.CONCLUSION: TIP30 plays an important role in predisposing hepatoblastoma cells to apoptosis through regulating expression levels of these genes. Ad-TIP30carrying exogenous TIP30-anti-tumor genes may be regarded as a potential candidate for the treatment of hepatocellular carcinoma.

Drug therapy for ulcerative colitis

Ulcerative colitis (UC) is an inflammatory destructive disease of the large intestine occurred usually in the rectum and lower part of the colon as well as the entire colon. Drug therapy is not the only choice for UC treatment and medical management should be as a comprehensive whole.Azulfidine, Asacol, Pentasa, Dipentum, and Rowasa all contain 5-aminosalicylic acid (5-ASA), which is the topical anti-inflammatory ingredient. Pentasa is more commonly used in treating Crohn's ileitis because Pentasa capsules release more 5-ASA into the small intestine than Asacol tablets. Pentasa can also be used for treating mild to moderate UC. Rowasa enemas are safe and effective in treating ulcerative proctitis and proctosigmoiditis. The sulfafree 5-ASA agents (Asacol, Pentasa, Dipentum and Rowasa) have fewer side effects than sulfa-containing Azulfidine. In UC patients with moderate to severe disease and in patients who failed to respond to 5-ASA compounds,systemic (oral) corticosteroids should be used. Systemic corticosteroids (prednisone, prednisolone, cortisone, etc.)are potent and fast-acting drugs for treating UC, Crohn's ileitis and ileocolitis. Systemic corticosteroids are not effective in maintaining remission in patients with UC.Serious side effects can result from prolonged corticosteroid treatment. To minimize side effects, corticosteroids should be gradually reduced as soon as the disease remission is achieved. In patients with corticosteroid-dependent or unresponsive to corticosteroid treatment, surgery or immunomodulator is considered. Immunomodulators used for treating severe UC include azathioprine/6-MP,methotrexate, and cyclosporine. Integrated traditional Chinese and Western medicine is safe and effective in maintaining remission in patients with UC.