原发性甲状旁腺功能亢进症患者术前病变甲状旁腺定位方法的评估

目的评价锝[^（99m）Tc]甲氧基异丁基异腈(99mTc-MIBI)显像及颈部超声对原发性甲状旁腺功能亢进症(primaryhyperparathyroidism,pHPT)患者在手术前进行病变甲状旁腺定位的敏感性和应用价值。方法对本院1983年6月到2002年6月期间收治的160例pHPT患者进行术前99mTc-MIBI显像检查和/或颈部超声检查。其中,男性53例(33.1%),女性107例(66.9%),平均年龄为(38.9±14.8)岁;100例接受99mTc-MIBI显像检查,148例进行高分辨率颈部超声检查。检查结果与手术所见和病理学进行比较。结果99mTc-MIBI显像和颈部超声对病变甲状旁腺定位的敏感性分别为94.0%和85.1%,阳性预测值分别为100%和89.1%,联合2种检查总的敏感性可达98.9%;在单个甲状旁腺病变组(n=145)中,99mTc-MIBI显像的敏感性为93.3%,颈部超声的敏感性为84.7%;甲状旁腺病变异位患者进行99mTc-MIBI显像的检测敏感性达100%,但是应用超声检测敏感性为56.3%。结论99mTc-MIBI显像和颈部超声对pHPT具有不同的敏感性,2种检查方法联合应用可以使敏感性增加。甲状旁腺异位对99mTc-MIBI显像的敏感性无影响,但是会使超声的敏感性下降,肿瘤大小影响超声检查敏感性,并且观察到99mTc-MIBI假阴性的各种原因,对于99mTc-MIBI检查阴性者应该排除可能的干扰因素。

脂肪乳剂所致的脂质过氧化及DNA损害

目的：了解静脉输注脂肪乳剂是否会增加机体的脂质过氧化和改变机体血浆维生素Ｅ浓度，脂肪乳剂所致的脂质过氧化是否会造成机体组织细胞的ＤＮＡ损害。 方法：２２ 例健康男性志愿者进行两个阶段的随机交叉研究。禁食一晚后于清晨８：００ 抽血作为基础对照，然后分别连续静脉输注２０％ Ｉｎｔｒａｌｉｐｉｄ 或２０％ Ｖａｓｏｌｉｐｉｄ ４ｈ（甘油三酯输注速度：０．１５ ｇ／ｋｇ·ｈ－ １），两种脂肪乳剂输注之间相隔２周，并分别于输注２ ｈ、４ ｈ 时抽血测定血浆脂质过氧化终产物ｍ ａｌｏｎａｌｄｅｈｙｄｅ（ＭＤＡ）和４－ｈｙｄｒｏｘｙａｌｋｅｎａｌ的总量；测定血浆α－ｔｏｃｏｐｈｅｒｏｌ代表维生素Ｅ浓度；自实验前一天起连续收集３ 天尿，检测ＤＮＡ 损害终产物——８－Ｈｙｄｒｏｘｙｇｕａｎｉｎｅ（８－ＯＨＧｕａ）量，作为ＤＮＡ 损害程度的分析。 结果：静脉输注脂肪乳剂过程中，血浆ＭＤＡ和４－ｈｙｄｒｏｘｙａｌｋｅｎａｌ明显增高，输注后２４ ｈ 降至输注前水平，两种脂肪乳剂之间无统计学上差异，脂肪乳剂输注过程中血浆维生素Ｅ浓度无明显变化。两种脂肪乳剂应用前后，尿中８－ＯＨＧｕａ含量无变化。 结论：静脉输注脂肪乳剂可增加机体脂质过氧化。

中链脂肪乳剂的临床应用

由于中链脂肪乳剂的结构、生化特征 ,具有氧化快而完全、较少依赖肉毒碱转运等优点 ,目前已普遍应用于临床营养支持。但是 ,临床上有关中链脂肪乳剂与传统的长链脂肪乳剂在各领域中应用的比较研究却较少 ,而且结果相差很大。本综述通过总结临床上有关中、长链脂肪乳剂的前瞻性随机对照研究资料 ,比较中。

结构甘油三酯与长链甘油三酯在健康志愿者中的代谢研究

目的:通过比较结构甘油三酯(STG)与长链甘油三酯(LCT)在健康志愿者中的代谢效应,以评估结构脂肪乳剂代谢特征。 方法:选择16例健康志愿者进行前瞻性随机交叉对照研究,采用高甘油三酯钳闭技术比较STG和LCT的代谢特征。 结果:STG的血浆清除率较LCT快1倍左右,STG组血清总游离脂肪酸、游离甘油及β-羟基丁酸盐浓度明显高于LCT组,血浆脂蛋白脂酶(LPL)活性明显大于LCT组并与所输注的甘油三酯量密切相关。 结论:健康志愿者血浆STG的清除率明显大于LCT,STG 的水解速度明显快于LCT。

Differentiation of rat marrow mesenchymal stem cells into pancreatic islet beta-cells

AIM: To explore the possibility of marrow mesenchymal stem cells (MSC) in vitro differentiating into functional isletlike cells and to test the diabetes therapeutic potency of Islet-like cells. METHODS: Rat MSCs were isolated from Wistar rats and cultured. Passaged MSCs were induced to differentiate into islet-like cells under following conditions: pre-induction with L-DMEM including 10 mmol/L nicotinamide+l mmol/L β-mercaptoethanol+200 mL/L fetal calf serum (FSC) for 24 h, followed by induction with serum free H-DMEM solution including 10 mmol/L nicotinamide+1 mmol/L,β-mercaptoethanol for 10 h. Differentiated cells were observed under inverse microscopy, insulin and nestin expressed in differentiated cells were detected with immunocytochemistry. Insulin excreted from differentiated cells was tested with radioimmunoassay. Rat diabetic models were made to test in vivo function of differentiated MSCs. RESULTS: Typical islet-like clustered cells were observed. Insulin mRNA and protein expressions were positive in differentiated cells, and nestin could be detected in predifferentiated cells. Insulin excreted from differentiated MSCs (446.93±102.28 IU/L) was much higher than that from pre-differentiated MSCs (2.45+0.81 IU/L (P<0.01). Injected differentiated MSCs cells could down-regulate glucose level in diabetic rats. CONCLUSION: Islet-like functional cells can be differentiated from marrow mesenchymal stem cells, which may be a new procedure for clinical diabetes stem -cell therapy, these cells can control blood glucose level in diabetic rats. MSCs may play an important role in diabetes therapy by islet differentiation and transplantation.

Clinical features,diagnosis,treatment and prognosis of multiple primary colorectal carcinoma

AIM: To investigate the clinical features, diagnosis,treatment and prognosis of multiple primary colorectal carcinomas (MPCC). METHODS: A retrospective analysis of 37 patients with MPCC from 1974 to 1998 was carried out.RESULTS: The incidence of MPCC was 2.74%(37/1 348) in patients with primary colorectal carcinomas, 15 cases of them were patients with synchronous carcinomas (SC) and 22 cases were diagnosed as metachronous carcinomas (MC). Most tumors were located in the right colon and rectum. Fifty-five percent (12/22) of MC were diagnosed within 3 years after tumor resection and 41%(9/22) of MC occurred after 8 years. Radical resections were performed in all patients except for 1 case. The 5-year survival rate of SC was 72.7%(8/11) and that of MC after the first cancer and second cancer was 71.4%(15/21) and 38.9%(7/18), respectively. CONCLUSION: The results indicate the importance of complete preoperative examination, careful intraoperative exploration and periodic postoperative surveillance. Early diagnosis and radical resection can increase survival rate of MPCC.

Constitutive activation of Stat3 signaling pathway in human colorectal carcinoma

AIM: Signal transducers and activators of transcription (STATs) are a family of transcription factors activated in response to cytokines and growth factors. Constitutive activation of Stat3 has been observed in a growing number of tumor-derived cell lines, as well as tumor specimens from human cancers. The purpose of this study was to investigate the expression of p-Stat3, activated form of Stat3, and its downstream mediators including cyclin D1 and Bcl-XL in colorectal carcinoma (CRC), and to explore the possible mechanism of Stat3 signaling pathway in the tumorigenesis of colorectal carcinoma. METHODS: Tissue samples from 45 patients of primary colorectal carcinoma were selected for studying Stat3 signaling pathway protein expression. Western blot analysis was used to measure the expression of p-Stat3, cyclin D1, and Bcl-xu proteins in colorectal carcinomas. Furthermore, the expression patterns of these proteins were analyzed for their distribution at the cellular level by immunohistochemical staining of the tissues. RESULTS: Protein levels of p-Stat3, cyclin D1, and Bcl-XL were increased in colorectal carcinomas compared with adjacent normal mucosae (P<0.05). Elevated levels of pStat3 were correlated with the nodal metastasis and the stage (P<0.05). Overexpression of cyclin D1 was associated with the nodal metastasis (P<0.05). There was also a significant correlation between the expressions of p-Stat3 and cyclin D1 (r=0.382, P<0.05). CONCLUSION: Constitutive activation of Stat3 may play an important role in the tumorigenesis of colorectal carcinoma, and the detailed mechanism of Stat3 signaling pathway in CRC deserves further investigation.

Heat shock protein 70 antisense oligonucleotide inhibits cell growth and induces apoptosis in human gastric cancer cell line SGC-7901

AIM: Heat shock protein (HSP)70 is over-expressed in human gastric cancer and plays an important role in the progression of this cancer. We investigated the effects of antisense HSP70 oligomer on human gastric cancer cell line SGC-7901, and its potential role in gene therapy for this cancer.METHODS: Human gastric cancer cell line SGC-7901 was treated in vitro with various concentrations of antisense HSP70 oligonucleotides at different intervals. Growth inhibition was determined as percentage by trypan blue dye exclusion test. Extracted DNA was electrophoresed on agarose gel, and distribution of cell cycle and kinetics of apoptosis induction were analyzed by propidium iodide DNA incorporation using flow cytometry, which was also used to detect the effects of antisense oligomer pretreatment on the subsequent apoptosis induced by heat shock in SGC-7901 cells. Proteins were extracted for simultaneous measurement of HSP70 expression level by SDS-PAGE Western blotting.RESULTS: The number of viable cells decreased in a doseand time-dependent manner, and ladder-like patterns of DNA fragments were observed in SGC-7901 cells treated with antisense HSP70 oligomers at a concentration of 10 μmol/L for 48 h or 8 μmol/L for 72 h, which were consistent with inter-nucleosomal DNA fragmentation. Flow cytometric analysis showed a dose- and time-dependent increase in apoptotic rate by HSP70 antisense oligomers. This response was accompanied with a decrease in the percentage of cells in the G1 and S phases of the cell cycle, suggesting inhibition of cell proliferation. In addition, flow cytometry also showed that pretreatment of SGC-7901 cells with HSP70 antisense oligomers enhanced the subsequent apoptosis induced by heat shock treatment. Western blotting demonstrated that HSP70 antisense oligomers inhibited HSP70 expression, which preceded apoptosis, and HSP70 was undetectable at the concentration of 10 μmol/L for 48 h or 8 μmol/L for 72 h.CONCLUSION: Antisense HSP70 oligomers can abrogate HSP70 expression in SGC-7901 cells, which may in turn induce apoptosis and inhibit cell proliferation, conversely suggesting that HSP70 is required for the proliferation and survival of human gastric cancer cells under normal conditions.

Gallbladder motor function, plasma cholecystokinin and cholecystokinin receptor of gallbladder in cholesterol stone patients

AIM: To study the interactive relationship of gallbladder motor function, plasma cholecystokinin (CCK) and cholecystokinin A receptor (CCK-R) of gallbladder in patients with cholesterol stone disease.METHODS: Gallbladder motility was studied by ultrasonography in 33 patients with gallbladder stone and 10 health subjects as controls. Plasma CCK concentration was measured by radioimmunoassay in fasting status (CCK-f) and in 30 min after lipid test meal (CCK-30).Radioligand method was employed to analyze the amount and activity of CCK-R from 33 gallstone patients having cholecystectomy and 8 persons without gallstone died of severe trauma as controls.RESULTS: The percentage of cholesterol in the gallstone composition was more than 70%. The cholesterol stone type was indicated for the patients with gallbladder stone in this study. Based on the criterion of gallbladder residual fraction of the control group, 33 gallstone patients were divided into two subgroups, contractor group (14 cases)and non-contractor group (19 cases), The concentration of CCK-30 was significantly higher in non-contractor group than that in both contractor group and control group (55.86±3.86 pmol/l vs 37.85±0.88 pmol/l and 37.95±0.74 pmol/L, P＜0.01), but there was no difference between contractor group and control group. Meanwhile no significant difference of the concentration of CCK-f could be observed among three groups. The amount of CCK-R was lower in non-contractor group than those in both control group and contractor group (10.27±0.94 fmol/mg vs24.59±2.39 fmol/mg and 22.66±0.55 fmol/mg, P＜0.01).The activity of CCK-R shown as KD in non-contractor group decreased compared to that in control group and contractor group. Only was the activity of CCK-R lower in contractor group than that in control group. The ejection fraction correlated closely with the amount of CCK-R (r = 0.9683,P＜0.01), and the concentration of CCK-30 correlated negatively with the amount of CCK-R closely (r = -0.9627,P＜0.01).CONCLUSION: The distinctive interactive relationship of gallbladder emptying, plasma CCK and CCK-R in gallbladder from this study suggested that the defect of CCK-R may be a key point leading to the impairment of gallbladder motor function and the pathogenesis of cholesterol gallstoneformation may differ in two subgroups of gallstone patient,gallbladder non-contractor group or contractor group.

Expressions of cysteine-rich61,connective tissue growth factor and Nov genes in hepatocellular carcinoma and their clinical significance

AIM: To investigate the expression of cysteine-rich61 (Cyr61),connective tissue growth factor (CTGF) and nephroblastomaoverexpressecl gene (Nov) in hepatocellular carcinoma (HCC),and to evaluate the relationship between Cyr61, CTGF and Nov genes expression with invasion and metastasis of HCC.METHODS: Thirty-one HCC specimens were divided into small hepatocellular carcinoma (SHCC), nodular hepatocellular carcinoma (NHCC), solitary large hepatocellular carcinoma (SLHCC) according to their diameter and number of nodes. Reverse transcription polymerse chain reaction (RT-PCR) was used to detect the mRNA expression levels of Cyr61, CTGF and Nov genes in 31 resected specimens of hepatocellular carcinoma and para-cancerous normal liver tissues semi-quantitatively and the relation between their expression levels and clinical pathological parameters were compared.RESULTS: The expressions of Cyr61 and CTGF mRNA in carcinoma tissues were significantly higher than those in para-cancerous normal liver tissues (P<0.01). The expressions of Cyr61 and CTGF mRNA in HCC with venous invasion were higher than those in HCC without venous invasion. CTGF expression in HCC Edmondson's grade Ⅲ-IV was significantly higher than that in HCC Edmondson's grade I-II (P = 0.022). There was no obvious correlation between Nov mRNA and clinical-pathological features.Compared to NHCC, SLHCC had better cell differentiation,easier capsule formation, less microscopic venous invasion,milder liver cirrhosis. The expressions of Cyr61 and CTGF mRNA in NHCC were significantly higher than those in SLHCC and SHCC.CONCLUSION: Cyr61 and CTGF genes may play an important role in hepatocellular carcinogenesis and correlate with recurrence and metastasis of hepatocellular carcinoma.SLHCC has better biological behaviors than NHCC.

Risk factors of pancreatic leakage after pancreaticoduodenectomy

AIM: To analyze the risk factors for pancreatic leakage after pancreaticoduodenectomy (PD) and to evaluate whether duct-to-mucosa pancreaticojejunostomy could reduce the risk of pancreatic leakage.METHODS: Sixty-two patients who underwent PD at our hospital between January 2000 and November 2003 were reviewed retrospectively. The primary diseases of the patients included pancreas cancer, ampullary cancer, bile duct cancer, islet cell cancer, duodenal cancer, chronic pancreatitis, pancreatic cystadenoma, and gastric cancer.Standard PD was performed for 25 cases, PD with extended lymphadenectomy for 27 cases, pylorus-preserving PD for 10 cases. A duct-to-mucosa pancreaticojejunostomy was performed for patients with a hard pancreas and a dilated pancreatic duct, and a traditional end-to-end invagination pancreaticojejunostomy for patients with a soft pancreas and a non-dilated duct. Patients were divided into two groups according to the incidence of postoperative pancreaticojejunal anastomotic leakage: 10 cases with leakage and 52 cases without leakage. Seven preoperative and six intraoperative risk factors with the potential to affect the incidence of pancreatic leakage were analyzed with SPSS10.0 software. Logistic regression was then used to determine the effect of multiple factors on pancreatic leakage.RESULTS: Of the 62 patients, 10 (16.13%) were identified as having pancreatic leakage after operation. Other major postoperative complications included delayed gastric emptying (eight patients), abdominal bleeding (four patients), abdominal abscess (three patients) and wound infection (two patients). The overall surgical morbidity was 43.5% (27/62). The hospital mortality in this series was 4.84% (3/62), and the mortality associated with pancreatic fistula was 10% (1/10). Sixteen cases underwent duct-to-mucosa pancreaticojejunostomy and 1 case (1/16, 6.25%) devel-oped postoperative pancreatic leakage, 46 cases underwent invagination pancreaticojejunostomy and 9 cases (9/46, 19.6%)developed postoperative pancreatic leakage. General risk factors including patient age, gender, history of jaundice,preoperative nutrition, pathological diagnosis and the length of postoperative stay were similar in the two groups.There was no statistical difference in the incidence of pancreatic leakage between the patients who received the prophylactic use of octreotide after surgery and the patients who did not undergo somatostatin therapy.Moreover, multivariate logistic regression analysis showed that none of the above factors seemed to be associated with pancreatic fistula. Two intraoperative risk factors,pancreatic duct size and texture of the remnant pancreas,were found to be significantly associated with pancreatic leakage. The incidence of pancreatic leakage was 4.88%in patients with a pancreatic duct size greater than or equal to 3 mm and was 38.1% in those with ducts smaller than 3 mm (P = 0.002). The pancreatic leakage rate was 2.94% in patients with a hard pancreas and was 32.1% in those with a soft pancreas (P = 0.004). Operative time,blood loss and type of resection were similar in the two patient groups. The incidence of pancreatic leakage was 6.25% (1/16) in patients with duct-to-mucosa anastomosis,and was 19.6% (9/46) in those with traditional invagination anastomosis. Although the difference of pancreatic leakage between the two groups was obvious, no statistical signific-ance was found. This may be due to the small number of patients with duct-to-mucosa anastomosis.By further analyzing with multivariate logistic regression,both pancreatic duct size and texture of the remnant pancreas were demonstrated to be independent risk factors (P= 0.007 and 0.017, OR = 11.87 and 15.45). Although anastomotic technique was not a significant factor,pancreatic leakage rate was much less in cases that underwent duct-to-mucosa pancreaticojejunostomy.CONCLUSION: Pancreatic duct size and texture of the remnant pancreas are risk factors influencing pancreatic leakage after PD. Duct-to-mum pancreaticojejunostomy,as a safe and useful anastomotic technique, can reduce pancreatic leakage rate after PD.

Clinical features and mismatch repair gene mutation screening in Chinese patients with hereditary nonpolyposis colorectal carcinoma

AIM: Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominantly- inherited cancer-susceptibility syndrome that confers an increased risk for colorectal cancer and a variety of other tumors at a young age. It has been associated with germline mutations in five mismatch repair (MMR) genes (hMSH2, hMLH1, hPMS1, hPMS2, and hMSH6/GTBP). The great majority of germline mutations were found in hMSH2 and hMLH1. The purpose of this study was to analyze the clinical features of Chinese HNPCC patients and to screen hMSH2 and hMLH1 gene mutations.METHODS: Twenty-eight independent Chinese families were collected, of which 15 met Amsterdam criteria I and 13 met the Japanese clinical diagnosis criteria. The data were recorded including sex, site of colorectal cancer (CRC),age of diagnosis, history of synchronous and/or metachronous CRC, instance of extracolonic cancers, and histopathology of tumors. Peripheral blood samples were collected from all pedigrees after formal written consents were signed. PCR and denaturing high-performance liquid chromatography (DHPLC) were used to screen the coding regions of hMSH2 and hMLH1 genes. The samples showing abnormal DHPLC profiles were sequenced by a 377 DNA sequencer.RESULTS: One hundred and seventy malignant neoplasms were found in one hundred and twenty-six patients (multiple cancer in twenty-three), including one hundred and twentyseven CRCs, fifteen gastric, seven endometrial, and five esophageal cancers. Seventy-seven point eight percent of the patients had CRCs, sharing the features of early occurrence (average age of onset, 45.9 years) and of the right-sided predominance reported in the literature. In Chinese HNPCC patients, gastric cancer occurred more frequently, accounting for 11.9% of all cancers patients and ranking second in the spectrum of HNPCC predisposing cancers. Synchronous CRCs occurred less frequently, only accounting for 3.1% of the total CRCs. Twenty percent of the colorectal patients had metachronous CRCs within 10 years after operation. Eight hMSH2 or hMLH1 gene sequence variations were found in twelve families, including the first Mongolian kindred with a hMSH2 gene mutation.CONCLUSION: HNPCC is characterized by an early-age onset, proximal predominance of CRC, multiple metachronous CRCs, and an excess of extra-colonic cancers. Frequent gastric cancer occurrence and less synchronous CRCs are the remarkable features in Chinese HNPCC patients. DHPLC is a powerful tool in hMSH2 and hMLH1 gene mutation screening, hMLH1 gene mutations, especially of the first nine exons, have been found more common than hMSH2 gene mutations in Chinese patients. Three of seven mutations have been found to be novel, and the germline G204X nonsense mutation in the third exon of hMSH2 has become the first MMR gene mutation found in Chinese Mongolian people.

Clinical characteristics and management of patients with early acute severe pancreatitis:Experience from a medical center in China

AIM:To study clinical characteristics and management of patients with early severe acute pancreatitis (ESAP).METHODS: Data of 297 patients with severe acute pancreatitis (SAP) admitted to our hospital within 72h after onset of symptoms from January 1991 to June 2003 were reviewed for the occurrence and development of early severe acute pancreatitis (ESAP).ESAP was defined as presence of organ dysfunction within 72 h after onset of symptoms.Sixtynine patients had ESAP, 228 patients without organ dysfunction within 72 h after onset of symptoms had SAR The clinical characteristics,incidence of organ dysfunction during hospitalization and prognosis between ESAP and SAP were compared.RESULTS.Impairment degree of pancreas (Balthazar CT class) in ESAP was more serious than that in SAP (5.31±0.68vs 3.68±0.29, P<0.01). ESAP had a higher mortality than SAP (43.4% vs 2.6%, P<0.01), and a higher incidence ofhypoxemia (85.5% vs 25%, P<0.01), pancreas infection (15.9% vs 7.5%, P<0.05),abdominal compartment syndrome (ACS) (78.3% vs 23.2%,P<0.01) and multiple organ dysfunction syndrome (MODS)(78.3% vs 10.1%,P<0.01).In multiple logistic regression analysis,the main predisposing factors to ESAP were higher APACHE II score, Balthazar CT class,MODS and hypoxemia.CONCLUSION:ESAP is characterised by MODS, severe pathological changes of pancreas,early hypoxemia and abdominal compartment syndrome. Given the poor prognosis of ESAP,these patients should be treated in specialized intensive care units with special measures such as close supervision, fluid resuscitation, improvement of hypoxemia,reduction of pancreatic secretion, elimination of inflammatory mediators,prevention and treatment of pancreatic infections.

Amelioration of hemodynamics and oxygen metabolism by continuous venovenous hemofiltration in experimental porcine pancreatitis

AIM: To investigate the potential role of continuous venovenous hemofiltration (CVVH) in hemodynamics and oxygen metabolism in pigs with severe acute pancreatitis (SAP).METHODS: SAP model was produced by intraductal injection of sodium taurocholate [4%, 1 mL/kg body weight (BW)] and trypsin (2 U/kg BW). Animals were allocated either to untreated controls as group 1 or to one of two treatment groups as group 2 receiving a low-volume CWH [20 mL/(kg·h)], and group 3 receiving a high-volume CVVH [100 (mL/kg·h)]. Swan-Ganz catheter was inserted during the operation. Heart rate, arterial blood pressure, cardiac output, mean pulmonary arterial pressure, pulmonary arterial wedge pressure, central venous pressure, systemic vascular resistance, oxygen delivery, oxygen consumption,oxygen extraction ratio, as well as survival of pigs were evaluated in the study.RESULTS: Survival time was significantly prolonged by low-volume and high-volume CWHs, which was more pronounced in the latter. High-volume CWH was significantly superior compared with less intensive treatment modalities (low-volume CWH) in systemic inflammatory reaction protection.The major hemodynamic finding was that pancreatitis-induced hypotension was significantly attenuated by intensive CVVH (87.4±12.5 kPa vs 116.3±7.8 kPa, P＜0.01). The development of hyperdynamic circulatory failure was simultaneously attenuated, as reflected by a limited increase in cardiac output,an attenuated decrease in systemic vascular resistance and an elevation in oxygen extraction ratio.CONCLUSION: CVVH blunts the pancreatitis-induced cardiovascular response and increases tissue oxygen extraction. The high-volume CWH is distinctly superior in preventing sepsis-related hemodynamic impairment.

Detection of micrometastasis in peripheral blood by multi-sampling in patients with colorectal cancer

AIM: To evaluate the reverse transcriptase-PCR assay and multiple sampling for detection of cytokeratin-positive cells in peripheral blood of colorectal carcinoma patients and to investigate the clinical significance of micrometastasis in peripheral blood.METHODS: The expression of CK20 mRNA by RT-PCR was investigated in bone marrow, portal vein and peripheral blood in 58 colorectal cancer patients and 12 controls without known cancer. The peripheral blood was sampled twice at intervals of 3 d before operation. All the patients were followed up for one year.RESULTS: There was no positive expression of CK20mRNA in 12 volunteers. The positive expression of CK20mRNA was 77.6% (45/58) in bone marrow, and that in portal vein was 74.1% (43/58) of colorectal carcinoma patients.The positive expression of CK20mRNA cells in peripheral blood rose from 44.8% (26/58) to 69.0% (40/58) (P＜0.01).The total positivity of CK20mRNA expression in peripheral blood was similar to the positivity of CK20mRNA in bone marrow and portal vein. The positive rates became higher in later clinical stages than in early stages. The CK20mRNA positive patients had a higher relapse rate within one year than the CK20mRNA negative patients.CONCLUSION: Multiple blood sampling can increase the detection of tumor cells in peripheral blood by RT-PCR for CK20mRNA in colorectal carcinoma patients and it is as sensitive and specific as that of bone marrow and portal vein. This technique may be reliable and convenient to diagnose micrometastasis of colorectal carcinoma and has an important significance in determining the prognosis of cancer patients.

Expression of MMP-2,TIMP-2 protein and the ratio of MMP-2/TIMP-2 in gallbladder carcinoma and their significance

AIM: To study the correlation between expression of MMP-2, TIMP-2 protein and the ratio of MMP-2/TIMP-2 and clinicalpathological parameters of patients with gallbladder carcinoma.METHODS: Carcinomas (n=45) and polypoid lesions (n=15) of the gallbladder were studied for the expression of MMP-2 and TIMP-2 protein by immunohistochemical avidin-biotin-complex method and image analysis. Clinicalpathological data of patients with gallbladder carcinoma such as histological type, grade of differentiation, level of infiltration, liver invasion and lymph node involvement, etc, were recorded.RESULTS: There was significant difference between the average level (1.123±0.108 VS 1.030±0.054, P=0.002) of MMP-2, the ratio (1.050±0.013 VS0.937±0.078, P=0.003) of MMP-2/TIMP-2 in gallbladder carcinomas and in polypoid lesions of the gallbladder. Significant difference was found between the expression of MMP-2 in early stage and advanced tumors, but there was no correlation between MMP-2 protein expression and histological type, differentiation degree, infiltration level, lymph node involvement or liver invasion. Although no difference was observed between TIMP-2 expression and histological type or differentiation degree, signific ant difference was found between TIMP-2 expression and different Nevin stage, infiltration level, local lymph node involvement or liver invasion (1.168±0.067 VS1.048±0.075, 1.170±0.062 vs 1.039±0.06g, 1.039±0.076 VS1.147±0.083, 1.048±0.074 vs 1.103±0.095, P＜0.05). MMP-2/TIMP-2 ratio did not correlate with histological type, grade of differentiation and liver invasion, but significant differences were found between MMP-2/TIMP-2 ratio and different Nevin stage, infiltration level and lymph node involvement in patients with carcinoma of gallbladder.CONCLUSION: TIMP-2 and MMP-2/TIMP-2 ratio could reflect more accurately biological characteristic of gallbladder carcinoma and MMP-2/TIMP-2 ratio might be a new significant marker in early diagnosis, in the judgment of invasion or metastasis and the estimate of prognosis in patients with gallbladder carcinomas.

Matrix metalloproteinase-2 and tissue inhibitor of metallo-proteinase-2 in colorectal carcinoma invasion and metastasis

AIM: To explore the relationship between matrix metalloProteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TTMP-2) in the development of colorectal carcinoma and to provide a valuable marker for clinical diagnosis.METHODS: Twenty-five patients with colorectal carcinoma underwent surgical resection. Samples were taken from tumor sites and normal tissues. MMP-2 activity was determined by gelatin zymography. Western blot and ABC immunohist ochemical staining were used to detect the expression levels of MMP-2 and TIMP-2 in normal and colorectal carcinoma tissues. Statistical analyses were performed using the Student's ttest and one-way ANOVA. P＜0.05 was considered statistically .significant. All the statistical analyses were performed using SPSS 10.0software. RESULTS: MMP-2 activity could be detected in both normal and colorectal carcinoma tissues. MMP-2 actMty in colorectal carcinoma tissues was much higher than that in normal tissues (P＜0.05, t =3.916, 4.227). MMP-2 adivity was positively related to the colorectal carcinoma invasion depth, lymph node metastasis and Duke's stage. Western blot and ABC immunohistochemical staining demonstrated that the expression level of MMP-2 in colorectal carcinoma tissues was much higher than that in normal tissues (P＜0.05, t = 9.429), but the expression level of TIMP-2 in colorectal carcinoma tissues was much lower than that in normal tissues (P＜0.05, t = 7.329). The MMP-2/TIMP-2 ratio of colorectal carcinoma was much higher than that of normal tissues. With the progression of invasion depth, lymph node metastasis and tumor Duke's stage, the activity and expression level of MMP-2 and TIMP-2 gradually increased, but the MMP-2/TIMP-2 ratio gradually decreased. CONCLUSION: The balance between MMP-2 and TIMP-2 plays a crucial role in the process of colorectal carcinoma invasion and metastasis.

Different approaches to caudate lobectomy with“curettage and aspiration”technique using a special instrument PMOD:A Report of 76 cases

AIM: To study different approaches to caudate lobectomy with "curettage and aspiration" technique using Peng's multifunctional operative dissector (PMOD). The surgical procedure of isolated complete caudate lobectomy was specially discussed.METHODS: In 76 cases of various types of caudate lobectomy, three approaches were used including left side approach, right side approach, and anterior approach.Among the 76 cases, isolated complete caudate lobectomy was carried out in 6 cases with transhepatic anterior approach. The surgical procedure consisted of mobilization of the total liver, ligation and separation of the short hepatic veins, splitting the liver parenchyma through the Cantlie's plane, ligation and division of the caudate portal triads from the hilum, dissection of the root of major hepatic veins,detachment of the caudate lobe from liver parenchyma.RESULTS: The mean operative time was 285±51 min,the mean blood loss was 1 600 mi. No severe complications were observed. Among the 6 cases receiving isolated complete caudate lobectomy with transhepatic anterior approach, one case died 17 months after operation due to disease recurrence and liver failure, the other 5 cases have been alive without recurrence, with one longest survival of 49 months.CONCLUSION: The choice of approach is essential to the success of caudate lobectomy. As PMOD and "curettage and aspiration" technique can delineate intrahepatic or extrahepatic vessels clearly, caudate lobe resection has become safer, easier and faster.

Clinical characteristics and prognostic factors of severe acute pancreatitis

AIM: To investigate the clinical characteristics and prognostic factors of a consecutive series of patients with severe acute pancreatitis (SAP). METHODS: Clinical data of SAP patients admitted to our hospital from January 2003 to January 2004 were retrospectively reviewed. Collected data included the age, gender, etiology, length of hospitalization, APACHE Ⅱ score at admission, local and organ/systemic complications of the patients. RESULTS: Of the 268 acute pancreatitis patients, 94 developed SAP. The mean age of SAP patients was 52 years, the commonest etiology was cholelithiasis (45.7%), the mean length of hospitalization was 70 d, the mean score of APACHE Ⅱ was 7.7. Fifty-four percent of the patients developed necrosis, 25% abscess, 58% organ/systemic failure. A total of 23.4% (22/94) of the SAP patients died. Respiratory failure was the most common organ dysfunction (90.9%) in deceased SAP patients, followed by cardiovascular failure (86.4%), renal failure (50.0%). In the SAP patients, 90.9% (20/22) developed multiple organ/systemic failures. There were significant differences in age, length of hospitalization, APACHE Ⅱ score and incidences of respiratory failure, renal failure, cardiovascular failure and hematological failure between deceased SAP patients and survived SAP patients. By multivariate logistic regression analysis, independent prognostic factors for mortality were respiratory failure, cardiovascular failure and renal failure. CONCLUSION: SAP patients are characterized by advanced age, high APACHE Ⅱ score, organ failure and their death is mainly due to multiple organ/systemic failures. In patients with SAP, respiratory, cardiovascular and renal failures can predict the fatal outcome and more attention should be paid to their clinical evaluation.

Survivin antisense compound inhibits proliferation and promotes apoptosis in liver cancer cells

AIM: To evaluate the effects of survivin on cell proliferation and apoptosis in liver cancer.METHODS: MTT assay was used to generate and optimize phosphorothioate antisense oligonucleotides (ODNs)LipofectamineTM2000 (LiP) compound by varying ODNs (μg):LiP (μL) ratios from 1:0.5 to 1:5. Then, liver cancer cells (HepG2) were transfected with the compound. By using RT-PCR and Western blot, the expression levels of survivin mRNA and proteins were detected in HepG2 cells treated with antisense compounds (ODNs:LiP = 1:4), and compared with those treated with sense compounds (1:4) as control.MTT assay was applied to the determination of cell proliferation in HepG2 cells. Active caspase-3 was evaluated by flow cytometric analysis. The morphological changes were assessed by electron microscopy. Laser scanning confocal microscopy was performed to detect the subcellular localization of survivin proteins in treated and untreated cells.RESULTS: Antisense compounds (1:4) down-regulated survivin expression (mRNA and protein) in a dose-dependent manner with an IC50 of 250 nmol/L. Its maximum effect was achieved at a concentration of 500 nmol/L, at whichmRNA and protein levels were down-regulated by 80%.The similar results were found in MTT assay. Antisense compound (1:4)-treated cells revealed increased caspase3-like protease activity compared with untreated cells.Untreated cells as control were primarily negative for the presence of active-caspase-3. As shown by transmission electron microscopy, treated cells with antisense compounds (1:4) resulted in morphological changes such as blebbing and loss of microvilli, vacuolization in the cytoplasm,condensation of the cytoplasm and nuclei, and fragmented chromatin. Immunofluorescence analysis confirmed the presence of survivin protein pool inside the cytoplasm in untreated cells. Labeled-FITC immunofluorescence staining of survivin clearly showed that survivin was distributed mainly in a spotted form inside the cytoplasm. Whereas cells treated with antisense compounds were rare and weak inside the cytoplasm.CONCLUSION: Down-regulation of survivin expression induced by the antisense compounds reduces tumor growth potential, promotes apoptosis and affects the localization of survivin proteins in HepG2 cells. Furthermore, survivin protein is a key molecule associated with proliferation and apoptosis, and antisense oligonucleotides targeting survivin have a bright prospect in the therapy of liver cancer.