Low expression of SLC22A1 is associated with a poor prognosis of hepatocellular carcinoma: analysis of 303 patients / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the association between SLC22A1 expression and the outcomes of hepatocellular carcinoma (HCC) patients.</p><p><b>METHODS</b>A tissue microarray of 303 HCC and matched adjacent noncancerous liver tissues (ANLTs) were constructed. The expression of SLC22A1 was tested by immunohistochemistry (IHC) and scored by two pathologists according to a 12-score scale (a score>6 was defined as high expression, and a score≤6 as low expression). The correlation of SLC22A1 expression with the clinicopathological features and the patients' outcome was analyzed.</p><p><b>RESULTS</b>All the ANLTs had a IHC score of 12, as compared to only 29 (9.6%) of the HCC tissues. The patients were divided into 2 groups based on the IHC scores: 59% (180/303) in low expression group and 41% (123/303) in high expression group. The disease-free survival (DFS) rates and overall survival (OS) rates were significantly lower in low SLC22A1 expression group than in the high expression group. The 1-, 3-, and 5-year DFS rates were 43%, 31% and 27% in the low expression group, and were 58%, 47% and 43% in the high expression group, respectively. The 1-, 3-, and 5-year OS rates were 66%, 38% and 32% in low expression group, and were 80%, 57% and 50% in the high expression group, respectively. A low expression of SLC22A1 was positively correlated with the tumor diameter, BCLC stage, tumor differentiation, and AFP levels (P<0.05), and was an independent predictor of poor overall survival (HR=1.454; 95% CI, 1.050-2.013).</p><p><b>CONCLUSIONS</b>Down-regulation of SLC22A1 is a malignant feature and a potential prognostic marker of HCC.</p>

Radiofrequency-assisted anterior approach right hepatectomy for hepatocellular carcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the safety and efficacy of radiofrequency-assisted anterior approach right hepatectomy for hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The clinic data of 12 HCC patients who underwent radiofrequency-assisted anterior approach right hepatectomy from January 2010 to July 2011 was analyzed retrospectively. Surgical techniques and treatment response were retrospectively reviewed. All the 12 patients were male, aging from 38 to 57 years with a mean of (48 ± 6) years. Ten of the 12 patients were infected with hepatitis B virus. A retrohepatic tunnel anterior to the surface of the inferior vena cava (IVC) was developed. The liver was hanged away from the IVC and radiofrequency was carried out along the Cantline's line. Scalpel was used to cut off the liver parenchyma along the middle of the ablated area until the parenchyma was fully resected. After short hepatic veins and the right hepatic vein were ligated, ligaments of right liver were fully isolated and right liver was resected. The t test was performed between 2 groups.</p><p><b>RESULTS</b>The surgical time was 165 to 295 minutes, with a mean of (230 ± 55) minutes. The bleeding was 150 to 1500 ml, with a mean of (516 ± 378) ml, which was better than those of anterior approach right hepatectomy ((1291 ± 1159) ml) and classical right hepatectomy ((2129 ± 2012) ml; t = 1.236, 3.265; P < 0.05). The postoperative hospital stay was 8 - 19 days, with a mean of (12 ± 4) days. There were no medical complications and no postoperative death. All patients were cured and discharged.</p><p><b>CONCLUSIONS</b>Radiofrequency-assisted anterior approach right hepatectomy for HCC is safe and effective and could effectively decrease intra-operative bleeding and shorten surgical time.</p>

Identification of Max binding protein as a novel binding protein of Nck1 and characterization of its role in inhibiting human liver cancer SK-HEP-1 cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The tendency of tumor cells to disperse throughout the liver is a distinct feature of hepatocellular carcinoma (HCC). Nck family adaptor proteins function to regulate actin cytoskeletal reorganization that leads to cell motility. We previously found that Max binding protein (MNT) was differentially expressed in HCC, and interacted with Nck1 by 2-DE. MNT is a protein member of the Myc/Max/Mad network which plays roles in cell proliferation, differentiation, and death. We investigated the effects of MNT on migration of human liver cancer SK-HEP-1 cells to study the migration regulatory role of MNT in HCC cells.</p><p><b>METHODS</b>Interaction between MNT and Nck1 was further validated in hepatoma cells by GST-pull down assay and immunoprecipitation. siRNAs specific to MNT (MNT siRNA) were used to knockdown MNT expression. Western blotting, transwell assay were used to determine the migration potential of cells.</p><p><b>RESULTS</b>Interaction between MNT and Nck1 was validated in hepatoma cells. MNT knockdown promoted the migration of human liver cancer SK-HEP-1 cells (P < 0.01).</p><p><b>CONCLUSION</b>The results suggest that MNT, via interaction with Nck1, inhibits hepatoma cell migration.</p>

Role of tissue factor in hepatocellular carcinoma genesis, invasion and metastasis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Numerous studies indicate that tissue factor (TF), namely tissue thromboplastin, has a close relationship with malignant tumor genesis and progress. It contributes to blood coagulation as well as the regulation of cellular differentiation, the formation of blood vessels, and also tumor recurrence and metastasis. The present study aimed to detect TF expression in hepatocellular carcinoma (HCC) patients and to elucidate its association with prognosis and clinical features of the disease.</p><p><b>METHODS</b>The plasma TF levels of 50 HCC patients and 30 controls were assayed by ELISA. The expressions of TF mRNA and protein in HCC tissues, adjacent tissues and normal tissues were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. The acquired data were analyzed with related clinic-pathological documents. The patients were followed up for five years, and the relationship between TF and prognosis was analyzed.</p><p><b>RESULTS</b>The plasma TF levels were significantly increased in HCC compared to the controls (P < 0.05), presenting a close relationship with differentiation level, tumor size and hepatocirrhosis occurrence (P < 0.05). There were remarkably higher values in cases of lymphatic metastasis, extrahepatic metastasis and portal tumor thrombus (PTT) (P < 0.05) compared to non-metastasis or non-tumor thrombus, but no significant difference with different focus number or envelope (P > 0.05). The positive rates and the relative expression of TF mRNA in HCC tissue were 63.0% (17/27) and 0.567 ± 0.268, respectively, significantly higher than that in adjacent tissues or normal tissues (P < 0.05). In the patients with positive results, the relative expression intensity varied significantly with different tumor size and index of local invasion and metastasis (P < 0.05). The positive rates and the relative expression intensities of TF protein in HCC tissue were 74.1% (20/27) and 4.093 ± 1.256, respectively, significantly higher than those in adjacent tissue or normal tissue (P < 0.05). In the patients with positive results, the relative expression intensity showed significant difference in different tumor size, differentiation level, and index of local invasion and metastasis (P < 0.05).</p><p><b>CONCLUSIONS</b>The TF levels were significantly higher in plasma and tissues of HCC patients, presenting a close relationship with the index of invasion and metastasis. It indicated that TF might be related to differentiation and metastasis of HCC.</p>

Combination with vascular resection and reconstruction in resection of hilar cholangiocarcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the value of vascular resection and reconstruction in resection of hilar cholangiocarcinoma.</p><p><b>METHODS</b>The clinical data of 17 patients with hilar cholangiocarcinoma received resection in combination with vascular resection and reconstruction from January 2000 to September 2009 was retrospectively analyzed. Among the 17 patients, 6 underwent portal vein segmental resection and end-to-end anastomosis, 3 underwent portal vein wedge resection, 1 underwent hepatic artery ligature, 2 underwent hepatic artery segmental resection and end-to-end anastomosis, 1 underwent portal vein arterialization, 1 underwent portal vein wedge resection and hepatic artery ligature simultaneously, 2 underwent portal vein segmental resection and hepatic artery segmental resection and end-to-end anastomosis simultaneously, 1 underwent portal vein segmental resection and right hepatic artery and gastroduodenal artery end-to-end anastomosis simultaneously.</p><p><b>RESULTS</b>Four patients died and the mortality was 4/17. Three patients died of renal dysfunction followed with multiple organ dysfunction and 1 patient died of sepsis shock. Among the 13 survive patients, 6 had a smooth postoperative recover and 7 developed complications: 3 had bile leakage, 1 had respiratory failure, 1 had cholangitis due to obstruction of U tube, 1 had abdominal infection and thrombosis in portal vein system and 1 had portal vein stenosis and liver abscess. Follow-up investigation showed that the median survival time was 18 months and four patients still alive.</p><p><b>CONCLUSIONS</b>Combination of vascular resection and reconstruction in the resection of hilar cholangiocarcinoma may help to improve the resection rate but still have a high postoperative risk. The complications of renal dysfunction should be alert during the postoperative observation. The procedure of hepatic arterial reconstruction may help to reduce postoperative morbidity.</p>

Long-term outcome and prognostic factors of intrahepatic cholangiocarcinoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The management of intrahepatic cholangiocarcinoma (ICC) remains a challenge due to poor prognosis. The aim of this study was to summarize the surgical management experience in recent 10 years and to identify the influencing factors related to outcome of patients with ICC in a single hepatobiliary center.</p><p><b>METHODS</b>From January 1995 to June 2005, 136 patients with ICC undergoing surgery were reviewed retrospectively. Survival rates of patients were calculated using the Kaplan-Meier method and compared by using the log-rank test. The prognostic factors were identified by the Cox regression model.</p><p><b>RESULTS</b>Seventy-nine of 136 patients underwent resection, and 65 of 79 patients were curative (R0). The surgical mortality was 2.2%. The 1-, 3- and 5-year survival rates of patients undergoing R0 resection were 72.1%, 35.6% and 20.1% respectively, which were significantly longer than those who underwent palliative resection and exploration, respectively (P < 0.01). At stage IV of the disease, 10 patients who underwent aggressive curative resection achieved a better median survival than those (n = 12) without resection (14 months vs 3 months, P < 0.001). The independent prognostic factors of the whole group were TNM stage (OR, 2.013, P = 0.008) and curative resection (OR, 2.957, P = 0.003). Higher TNM stage (OR, 1.894, P = 0.004) and lymph node metastasis (OR, 4.248, P = 0.005) linked to poor prognosis after R0 resection. For patients without lymph node metastasis, the median survival of those who underwent regional lymphadenectomy was comparable with those who did not (18 months vs 22 months, P = 0.817).</p><p><b>CONCLUSIONS</b>R0 resection is mandatory for ICC patient to achieve long-term survival. Aggressive resection benefits for selected patients with local advanced disease. Higher TNM stage and lymph node metastasis were poor prognostic factors for ICC patients after R0 resection.</p>

Combined hepatic resection and intraoperative thermal ablation for multifocal hepatocellular carcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the safety and efficacy of hepatic resection combined with intraoperative ablation to treat multifocal hepatocellular carcinoma.</p><p><b>METHODS</b>Clinical data of patients diagnosed with multifocal hepatocellular carcinoma and treated with hepatic resection combined with intraoperative ablation from March 1998 to September 2007 were retrospectively reviewed. Treatment response, postoperative complications and survival data were analyzed.</p><p><b>RESULTS</b>Combined treatment modalities were well tolerated except one patient dying of postoperative hepatic functional failure. The postoperative complication rate was 23.5% with a mortality rate of 6.7%. Postoperative complication included wound infection (1 case), bile leakage (1 case), subphrenic and pleural effusion (1 case), ablation-associated liver abscess (1 case), all of which were treated with non-surgical methods. The median survival time was 25.9 months. The 1, 3, 5 year survival rates were 70.6% (12/17), 23.5% (4/17), 17.6% (3/17), respectively. Three patients survived more than 5 years after surgery. Up to April 2008, 4 patients were still alive.</p><p><b>CONCLUSION</b>Hepatectomy combined with intraoperative thermal ablation provides a treatment modality for patients with multifocal hepatocellular carcinoma and may improve the prognosis.</p>

The impact of preoperative biliary drainage on surgical morbidity in hilar cholangiocarcinoma patients / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the impact of preoperative biliary drainage on surgical morbidity in hilar cholangiocarcinoma patients underwent surgery.</p><p><b>METHODS</b>One hundred and eleven consecutive patients with hilar cholangiocarcinoma whose serum total bilirubin (TBIL) level > 85 micromol/L and underwent surgery in the period from June 1998 to August 2007 were enrolled. There were 67 male and 44 female patients, aged from 26 to 82 years old with a mean of 56 years old.</p><p><b>RESULTS</b>Fifty-five patients underwent preoperative biliary drainage with a mean of 11.4 d of drainage period (drainage group), the other (n = 56) were the non-drainage group. The preoperative TBIL level of drainage group was (154 +/- 69) micromol/L, which was significantly lower than the value of pre-drainage (256 +/- 136) micromol/L (P = 0.000) and the value of non-drainage group (268 +/- 174) micromol/L (P = 0.005). ALT and GGT levels could be lowered by preoperative biliary drainage. The postoperative complications of these two groups were comparable (36.3% vs. 28.6%, P = 0.381). Four patients in drainage group and 5 patients in non-drainage group died of liver failure. Multivariate logistic regression indicated that hepatectomy (OR = 0.284, P = 0.003) was the independent risk factor associated with postoperative morbidity. Bismuth-Corlette classification (OR = 0.211, P = 0.028) was the independent risk factor linked to postoperative mortality.</p><p><b>CONCLUSIONS</b>Preoperative biliary drainage could alleviate liver injury due to hyperbilirubin, but it could not decrease the surgical morbidity and postoperative mortality. Concomitant hepatectomy and Bismuth-Corlette classification were independent risk factors linked to surgical risks.</p>

Changes in TGF-beta1/Smads signaling pathway in rats with chemical hepatocarcinogenesis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the changes in transforming growth factor beta 1 (TGF-beta1)/Smads signaling pathway in rats with chemical hepatocarcinogenesis.</p><p><b>METHODS</b>Fresh diethylnitrosamine (DENA) solution was administered in SD rats to induce hepatocellular carcinoma (HCC). The protein expressions of TGF-beta1, phosphorylated Smad2, Smad4 and Smad7 were detected in these rats with immunohistochemistry, and the mRNA expression of Smad4 was evaluated with RT-PCR.</p><p><b>RESULTS</b>Cirrhotic nodules occurred in the rats 8 weeks after DENA treatment, and HCC nodules were found 16 weeks after the treatment. In the normal liver tissue, very low levels of TGF-beta1 and Smad4 expressions, low Smad7 expression and high phosphorylated Smad2 expression were detected. The development of liver cirrhosis was accompanied by increased expressions of TGF-beta1, Smad4 and Smad7 but at 8 weeks after DENA treatment, the expression of phosphorylated Smad2 was significantly decreased, followed then by gradual increment till nearly the normal level. Twenty-two weeks after DENA treatment, Smad4 expression in liver tissue decreased markedly as compared with the levels at 8 and 16 weeks. The expressions of Smad4 and phosphorylated Smad2 in the HCC tissue was significantly lower than those in normal liver tissue.</p><p><b>CONCLUSION</b>Hepatocarcinogenesis involves very complex mechanisms, can can be related partially to the decreased Smad4 and phosphorylated Smad2 expression and TGFbeta1 and Smad7 overexpression in advanced stage of liver cirrhosis.</p>

A survey of bile duct injuries sustained during laparoscopic cholecystectomy / 中华外科杂志

<p><b>OBJECTIVE</b>To summarize the reasons for bile duct injury (BDI) after laparoscopic cholecystectomy (LC), and to determine the effect of multiple treatment after BDI.</p><p><b>METHODS</b>A retrospective cohort study was performed. The medical records of 110 patients diagnosed with BDI after LC from October 1993 to November 2007, in ten large hospitals in Guangdong of China, were reviewed.</p><p><b>RESULTS</b>Among 110 patients with BDI, 58 cases (52.7%) were local patients, whereas 52 cases (47.3%) were transferred from outside hospitals. Reasons for BDI following LC were: (1) Lack of experience of the LC operator (48.2%); (2) LC performed during acute cholecystitis (20.0%); (3) The structure of Calot triangle was unclear (15.5%); (4) Variable anatomical position (11.8%); (5) Intra-operation bleeding (4.5%). The commonest sites of injury were the choledochus and common hepatic duct (76.4%). Following BDI, endoscopic stenting or operative repair was performed in 106 patients. The overall success rate was 95.3% (101/106), with a mortality rate was 0.9% (1/106). Cholangitis occurred in 3.8% (4/106) cases. Choledocho-enterostomy operation was performed in almost 60.0% (63/106) cases, and the success rate was 93.7% (59/63). Endoscopic stenting or operative repair was performed immediately following BDI in 23.6% (25/106) patients, the success rate was 100%; and within 30 days in 63.2% (67/106) patients. Eighty-eight out of 106 patients who underwent repair were successful following the first operative procedure.</p><p><b>CONCLUSIONS</b>Factors such as an un-experienced operator and unclear anatomical position were causes of BDI following LC. Early operative repair should be regarded as the treatment of choice, in patients diagnosed with BDI. Early refer to an experienced hepatobiliary operator ensures a high success rate.</p>

Efficacy of postoperative chemotherapy combined with immunotherapy for hepatocellular carcinoma with major portal vein tumor thrombus / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of surgical treatment combined with immunochemotherapy via portal vein for hepatocellular carcinoma (HCC) with major portal vein tumor thrombus (PVTT).</p><p><b>METHODS</b>Between January 2001 and December 2005 76 HCC patients with tumor thrombus in portal trunk and (or) the first-order branch were recruited into the study. Patients were divided into group A (n = 29) and B (n = 47). Patients in group A were treated with hepatectomy plus portal thrombectomy in combination with postoperative adjuvant immunochemotherapy administered via portal vein. The immunochemotherapy regimen consisted of 5-Fluorouracil, Adriamycin, platinol and alpha-Interferon (PIAF). Patients in group B were subjected to hepatectomy plus thrombectomy alone. Survival rates were compared between two groups, and prognostic factors were identified.</p><p><b>RESULTS</b>Half-, One-, two- and three-year cumulative survival rates were markedly greater in group A than group B, being 82.3% vs 52.7%, 46.5% vs 20.2%, 14.3% vs 5.8%, 14.3% vs 5.8%, respectively. Group A had a significantly longer median survival time and median tumor-free survival time as compared with group B, being 11.5 months vs 6.0 months (P = 0.010), 4.5 months vs 2.4 months (P = 0.032), respectively. Multivariate analysis revealed that immunochemotherapy, pathological grading and tumor size were independent factors for survival times. And immunochemotherapy and pathological grading were independent factors for tumor-free survival time.</p><p><b>CONCLUSIONS</b>Surgical resection combined with adjuvant immunochemotherapy via portal vein represents as an effective modality for HCC with PVTT.</p>

The timing of hepatectomy for hepatolithiasis complicated with acute cholangitis / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the optimal timing of hepatectomy for intrahepatic lithiasis complicated with acute cholangitis.</p><p><b>METHODS</b>One hundred and twenty-six patients with hepatolithiasis who had a history of acute cholangitis and underwent hepatectomy were reviewed retrospectively. According to the period between the surgery and last attack of acute cholangitis, 126 patients were divided into 3 groups: > 3 months (group A, n = 73), 1 approximately 3 months (group B, n = 28), < 1 month (group C, n = 25). The operation time, blood loss, hospital stay, postoperative complications and stone residual rate were compared among the groups.</p><p><b>RESULTS</b>The intraoperative blood loss of C group was (644.0 +/- 625.7) ml, which was significantly higher than those of A and B group [(409.2 +/- 250.7) ml and (423.2 +/- 237.1) ml, respectively]. The numbers of patients who needed transfusion and the amount of blood transfusion in group C were also higher than those of group A and B. The incidence rate of complications, residual stone in group C were all markedly higher than those of group A and B. The period of hospital stay in group C was much longer than that in group A and B.</p><p><b>CONCLUSIONS</b>The optimal timing of hepatectomy for hepatolithiasis complicated with acute cholangitis is at least one month after subsidence of cholangitis.</p>

Surgical treatment for 68 patients with Caroli's disease / 中华外科杂志

<p><b>OBJECTIVE</b>To retrospectively investigate the diagnosis and the outcome of Caroli's disease treated by surgical procedures.</p><p><b>METHODS</b>The clinical data of 68 patients with Caroli's disease treated by surgical procedures between 1996 and 2002 were reviewed, retrospectively.</p><p><b>RESULTS</b>The patients, with a M/F ratio of 1:1.35 and a mean age of 46, presented mainly with recurrent cholangitis. Of all the patients, 26 had a history of operation for cholelithiasis or cholangitis. On admission, the image investigations suggested that the lesions located at left lobe in 44 patients, right lobe in 9 patients, and whole liver in 15 patients. The coexisting cyst in common bile duct was found in 20 patients. The malignant transformation was found in 5 patients (8.8%). Hepatectomy was performed in 82.4% of patients, with a morbidity rate of 15.0% and mortality rate of 0 after the surgery. The long-term outcome of symptom-free in hepatectomy group was 90.2%, significantly higher than the 33.3% in non-hepatectomy group (P < 0.01) after a 3 to 10 years of follow-up.</p><p><b>CONCLUSIONS</b>Hepatectomy offers a curative procedure for local Caroli's disease, and liver transplantation is a good option for diffuse sufferers.</p>

Experience of surgical resection of 103 hilar cholangiocarcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To summarize the experience of surgical resection of 103 hilar cholangiocarcinoma.</p><p><b>METHODS</b>One hundred and three consecutive cases of hilar cholangiocarcinoma who underwent surgical resection at our hospital over the past ten years were reviewed retrospectively. The clinical data and long-term outcome were analyzed.</p><p><b>RESULTS</b>Out of 103 cases, 43 patients underwent radical resection (41.7%), and 60 patients underwent palliative resection. There were 34 patients developed postoperative complications and 8 patients died in hospital. For the radical resection group, the median survival time was 29.9 months and 1-year, 3-year, 5-year survival rate was 69.6%, 42.0%, 20.9%, respectively, which was significant greater than 34.1%, 10.2%, 0 of the palliative resection group (P < 0.05). Over the past five years, 42 cases underwent pre-operative drainage of bile and the rate of combined liver resection reached 53.8%. The tumor radical resection rate has increased to 45.7%, the median survival time have reached 24.7 months (P < 0.05).</p><p><b>CONCLUSIONS</b>Improvement of pre-operative management, intraoperative pathology for resection margin, and combined liver resection may help in increasing the radical resection rate. Radical resection can improve postoperative survival, and produce a satisfactory outcome for patient with hepatic hilar cholangiocarcinoma.</p>

Use of autologous tumor vaccine in preventing recurrence of hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the effects of autologous tumor vaccines in preventing recurrences of hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>From March 1999 to June 2003, 80 patients with HCC undergoing resections were randomly assigned into a tumor vaccine group (n=40) and a control group (n=40). Tumor vaccines, consisting of formalin-fixed HCC tissue fragments, biodegradable sustained-releasers of granulocyte-macrophage-colony stimulating factor, interleukin-2, and an adjuvant, were developed. Every vaccine group patient received 3 vaccinations at a 2-week interval and the control group just received the adjuvant. Delayed-type-hypersensitivity (DTH) test and recurrent rates were analyzed.</p><p><b>RESULTS</b>Eight patients of the vaccine group and five patients of the control group were lost in the follow-up. Thirty-two patients completed the tumor vaccine procedure and no essential adverse effects occurred. 23/32 patients developed DTH responses against the fragments of HCC. The follow-up averaged 34.3 months (from 15 to 55 months). 1-, 2-, 3-year recurrence rates of the vaccine group were 12.6%, 35.9% and 54.0%, respectively; 1-, 2-, 3-year recurrence rates of the control group were 31.6%, 61.3% and 72.1%, respectively. The recurrent rate was significantly better in the tumor vaccine group than in the control group (P = 0.037).</p><p><b>CONCLUSIONS</b>Autologous tumor vaccine is a promising adjunctive modality to prevent recurrence of human HCC.</p>