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Infiltrating immune cells in the tumor microenvironment (TME) influence tumor progression and patient prognosis, making them attractive therapeutic targets for immunotherapy research. A deeper understanding of immune cell distributions in the TME in hepatocellular carcinoma (HCC) is needed to identify interactions among different immune cell types that might impact the effectiveness of potential immunotherapies. We performed multiplex immunohistochemistry using a tissue microarray of samples from 302 patients with HCC to elucidate the spatial distributions of immune cell subpopulations (CD3+, CD4+, CD8+, CD66b+, and CD68+) in HCC and normal liver tissues. We analyzed the associations between different immune subpopulations using Pearson's correlation. G(r) functions, K(r) functions and Euclidean distance were applied to characterize the bivariate distribution patterns among the immune cell types. Cox regression and Kaplan‐Meier analysis were used to evaluate the associations between tumor infiltration by different immune cells and patient outcomes after curative surgery. We also analyzed the relationship between the spatial distribution of different immune cell subpopulations with HCC patient prognosis. We found that the immune cell spatial distribution in the HCC TME is heterogeneous. Our study provides a theoretical basis for HCC immunotherapy. [ABSTRACT FROM AUTHOR]

Previous studies have indicated that complex interactions among viral, environmental and genetic factors lead to hepatocellular carcinoma (HCC). To identify susceptibility alleles for hepatitis B virus (HBV)-related HCC, the present study conducted a pilot two-phase genome-wide association study (GWAS) in 660 Han Chinese individuals. In phase 1, a total of 500,447 single-nucleotide polymorphisms (SNPs) were genotyped in 50 HCC cases and 50 controls using Affymetrix GeneChip 500k Array Set. In phase 2, 1,152 SNPs were selected from phase 1 and genotyped in 282 cases and 278 controls using the Illumina GoldenGate platform. The prior probability of HCC in control subjects was assigned at 0.01, and false-positive report probability (FPRP) was utilized to evaluate the statistical significance. In phase 1, one SNP (rs2212522) showed a significant association with HCC (P allele =5.23×10 -8 ; OR allele =4.96; 95% CI, 2.72-9.03). In phase 2, among 27 SNPs with unadjusted P allele <0.05, 9 SNPs were associated with HCC based on FPRP criteria (FPRP <0.20). The strongest statistical evidence for an association signal was with rs2120243 (combined OR allele =1.76; 95% CI, 1.39-2.22; P=2.00×10 -6 ), which maps within the fourth intron of VEPH1 . The second strongest statistical evidence for an association was identified for rs1350171 (combined OR allele =1.66; 95% CI, 1.33-2.07; P=6.48×10 -6 ), which maps to the region downstream of the FZD4 gene. The other potential susceptibility genes included PCDH9 , PRMT6 , LHX1 , KIF2B and L3MBTL4 . In conclusion, this pilot two-phase GWAS provides the evidence for the existence of common susceptibility loci for HCC. These genes involved various signaling pathways, including those associated with transforming growth factor β, insulin/phosphoinositide 3 kinase, Wnt and epidermal growth factor receptor. These associations must be replicated and validated in larger studies.

Background: Focal nodular hyperplasia (FNH), the second most common benign hepatic tumor after hemangioma, is characterized by a stellate central scar and hyperplastic nodules. Although some large FNH may be associated with significant symptoms, more frequently they are discovered incidentally on physical examination or the work-up of unrelated symptoms. Since its nature and pathogenesis are still controversial, accurate diagnosis of FNH based on clinical presentation and radiographic studies is difficult. The purpose of this study was to explore the diagnosis and treatment of FNH., Methods: Eighty-six FNH patients confirmed pathologically were treated at the Liver Cancer Institute in our hospital from 1996 to 2006. Their clinical manifestations, imaging presentation, pathological findings, and surgical results were analyzed retrospectively., Results: Of the 86 patients with 99 foci, 54 were male and 32 female, with a mean age of 37 years. Eighty patients had a single solitary focus and 6 had multiple foci. Tumor diameter was less than 5 cm in 69 patients, 5-10 cm in 15, and more than 10 cm in 2. The overall rate of correct preoperative diagnosis was 59.3% (51/86) including 32.9% (26/79) by color Doppler flow imaging (CDFI), 60.3% (35/58) by CT, and 77.4% (24/31) by MRI. All the 86 patients underwent resection with good curative effect., Conclusions: CT and MRI are important diagnostic methods for FNH but it is difficult to make a definite preoperative diagnosis for partial classical and all non-classical FNH patients. We suggest that patients with clinical symptoms or with indefinite diagnosis should accept surgical removal.

Objective: To investigate the effects of rapamycin (RPM) in inhibiting the growth and metastasis of hepatocellular carcinoma (HCC)., Methods: Human HCC cells of the line MHCC97H with a high potential of metastasis were divided into 3 groups to be cultured with cyclosporine A (CsA) 100 ng/ml, RPM 10 ng/ml, or CsA + RPM for 48 hours. Flow cytometry was used to examine the apoptosis and cell cycle MTT method was used to examine the effect of RMP on the proliferation of the MHCC97H cells. RT-PCR was used to detect the mRNA expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hypoxia-inducible factor-1alpha (HIF-1alpha), and transforming growth factor b (TGFb). Another MHCC97H cells were cultured in complete medium without RPM for 48 hours, then the protein expression of VEGF in the supernatant was detected by ELISA. Twenty-eight nude LCI-D20 mice were inoculated with human HCC cells and then divided into 4 groups to be fed with CsA (25 mg/kg), RPM (2 mg/kg), CsA + RPM, and normal saline (0.2 ml, as control group) for 35 days. Then the mice were killed to take the weight of inoculated tumor, measure the blood drug concentration, calculate the lung metastasis rate and number of metastatic foci, and observe pathology of the lung., Results: CsA showed no effect on the cycle of the MHCC97H cells. The MHCC97H cells of the RPM and CsA + RPM groups arrested at the stage G(0)/G(1) (both P = 0.000). MMT method also showed that the proliferation of the MHCC97H cells in the RPM and CsA + RPM groups were blocked (P = 0.003 and P = 0.002). However, CsA did not influence the proliferation of the MHCC97H cells. Flow cytometry showed that RPM did not promote the apoptosis of the MHCC97H cells. RT-PCR showed that RPM down-regulated the mRNA expression of VEGF and HIF-1alpha (both P < 0.05), however, did not influence the mRNA expression of bFGF, TGFb, and TGFb. The VEGF protein level in the supernatant of the culture fluid of MHCC97H cells of the RPM group was (890.3 +/- 25.1) pg/ml, significantly lower than that of the control group, (1583.7 +/- 17.3) pg/ml (P = 0.000). The tumor inhibiting rate of the RPM group was 63.7%, not significantly different from that of the RPM + CsA group (80.9%, P = 1.000). The metastatic rate of the CsA and control groups were both 100% with a higher number of metastatic tumors in the CsA group (P = 0.046)., Conclusion: RPM significantly inhibits the growth and metastasis of HCC. RPM-based immunosuppressive regimen may be of value in HCC patients receiving liver transplantation.

Aim: To report a retrospective analysis of preliminary results of 36 patients who received sirolimus (SRL, Rapamune, rapamycin) in a consecutive cohort of 248 liver allograft recipients., Methods: Thirty-six liver transplant patients with hepatocellular carcinoma (HCC) who were switched to SRL-based immunosuppression therapy from tacrolimus were enrolled in this study. The patients who were diagnosed as advanced HCC before orthotopic liver transplantation (OLT) were divided into group A (n = 11), those who were found to have HCC recurrence and/or metastasis after OLT were assigned to group B (n = 18), and those who developed renal insufficiency caused by calcineurin inhibitor (CNI) were assigned to group C (n = 7) after OLT., Results: The patients were followed up for a median of 10.4 mo (range, 3.8-19.1 mo) after conversion to SRL therapy and 12.3 mo (range, 5.1-34.4 mo) after OLT. Three patients developed mild acute cellular rejection 2 wk after initiating SRL therapy, which was fully reversed after prednisolone pulse therapy. In group A, only 1 patient was found to have HCC recurrence and metastasis 12 mo after OLT. In group B, 66.7% (12/18) patients (2 with progressive tumor, 7 with stable tumor and 3 without tumor) were still alive due to conversing to SRL and/or resection for HCC recurrence at the end of a median follow-up of 6.8 mo post conversion and 10.7 mo posttransplant. In group C, no HCC recurrence was demonstrated in 7 patients, and renal function became normal after SRL therapy. Thrombocytopenia (n = 2), anemia (n = 8), and oral aphthous ulcers (n = 7) found in our cohort were easily manageable., Conclusion: The conversion to SRL-based immunosuppression may inhibit the recurrence and metastasis of HCC and improve CNI-induced renal insufficiency in OLT patients with HCC.

Objective: To evaluate the effects of different selection criteria on the prognosis of hepatocellular carcinoma (HCC) patients undergoing liver transplantation (LT) and to explore the new criteria adapted for Chinese National Situation., Methods: A retrospective analysis was performed on 251 consecutive patients with HCC who underwent LT between April 2001 and January 2006 at our institution. We compared the outcome of the patients meeting different criteria such as milan, UCSF and the Pittsburgh modified TNM criteria. Survival rates were calculated using the Kaplan-Meier method, and differences between the curves were assessed by log-rank test., Results: There was no significant difference in 1, 2, 3-year survival rates and recurrence-free survival rates between milan criteria (n = 93; 86%, 77%, 77% and 91%, 86%, 86%) and UCSF criteria (n = 131; 90%, 83%, 83% and 92%, 89%, 89%). According to Pittsburgh criteria (n = 207), the 1, 2, 3-year survival rates and recurrence-free survival rates were 84%, 74%, 67% and 85%, 83%, 73%, respectively. For advanced tumors (over all the criteria), the survival rates and recurrence-free survival rates decreased significantly (n = 44; 65%, 43%, 43% and 47%, 43%, 43% respectively). When criteria (named as Shanghai Fudan Criteria) were expanded to HCC patients with solitary lesions < or = 9 cm in diameter, or no more than 3 lesions, the largest < or = 5 cm, with a total tumor diameter < or = 9 cm, there was no significant difference in 1, 2, 3-year survival rates and recurrence-free survival rates (n = 151; 88%, 80%, 80% and 90%, 88%, 88%, respectively) as compared with milan criteria, but more patients using Shanghai Fudan Criteria could undergo liver transplantation., Conclusion: Shanghai Fudan Criteria, which expanded the tumor size limits, does not adversely impact survival of HCC patients after LT.

Objective: To analyze the risk factors influencing the prognosis of orthotopic liver transplantation for hepatocellular carcinoma (HCC) and sum up the relevant clinical experience in diagnosis and treatment of HCC., Methods: The clinical data of 198 HCC patients, 177 males and 21 females, aged 49 (24-83), were analyzed., Results: The 0.5-, 1-, and 2-year survival rates were 89%, 78%, and 65 respectively. The rates of disease-free survival (DFS) were 85%, 73, and 67% respectively. Univariate analysis revealed that tumor size, presence of vascular invasion, Edmondson grade, TNM classification, and preoperative alpha-fetoprotein (AFP) were significantly related to DFS, and the 4 foregoing factors were also related to the survival rate. Cox regression analysis suggested that presence of vascular invasion was an independent prognostic factor of survival rate and DFS., Conclusion: Vascular invasion plays a leading role in evaluating the prognosis of orthotopic liver transplantation for HCC. It is important to discover the micro-metastasis and explore more effective approaches to prevent recurrence after transplantation.

Objective: To investigate the measures to further improve the therapeutic efficacy of liver transplantation., Methods: The clinical data of 203 consecutive cases of orthotopic liver transplantation (OLT) performed in Zhongshan Hospital, Fudan University April 2001 to October 2004, with the indications for considering OLT of hepatocellular carcinoma (HCC) (142 cases), liver cirrhosis (36 cases), fulminant hepatic failure (7 cases), Wilson's disease (6 cases), and other end-stage liver diseases (12 cases), including 199 cases of cadaveric OLT, inclusive of 2 cases of reduced-size and 1 case of splitting liver transplantation, and 4 cases of living-donor liver transplantation, were retrospectively analyzed. Multivariate analysis using Cox proportional hazards regression model was applied to determine the risk factors predicting liver transplantation prognosis for HCC., Results: For the whole group of the 201 patients, the 1-year and 2-year cumulative survival rates were 85.0% and 82.4% respectively and the rejection rate was 12.3%. In the HCC group the 1- and 2-year cumulative survival rates were 80.2% and 78.4%, and the 1- and 2-year disease-free survival (DFS) rates were 85.3% and 80.3% respectively. HCC recurrence was observed in 20 patients after OLT with a recurrence rate of 14.1%. Multivariate analysis revealed that the tumor size and portal vein tumor thrombus were the most independent and statistically significant factors affecting the DFS., Conclusion: OLT may be the most effective treatment option for patients with end-stage liver diseases and may also provide the opportunity of curative treatment or survival improvement for selected patients with hepatic malignancies.

Objective: To explore the diagnosis and management of early hepatic artery thrombosis (HAT) after liver transplantation., Methods: Routine examination of Color Doppler Imagine (CDI) was used to detect hepatic artery flow after liver transplantation in 220 cases from April 2001 to November 2004. Suspected patients were further confirmed by immediate hepatic artery angiography, and continuous infusion of urokinase through hepatic artery with catheter was performed to the patients with HAT., Results: HAT was identified in 6 patients (2.7%), occurring 5.5 days (2 - 19 days) after liver transplantation. Hepatic artery recanalization was obtained in 6 cases. One patient died from lung infection 2 months after liver transplantation. One patient underwent the second liver transplantation because of the recurrence of HAT 6 months after the first transplantation, but died from multiple system organ failure 2 months after the operation. The other 4 cases have been surviving well disease-freely., Conclusions: CDI is useful for the diagnosis of early HAT after liver transplantation. Contributes to recovery of the hepatic artery flow, continuous infusion of urokinase through hepatic artery with catheter may be the first choice of the therapy for early HAT.

Background: Selection of patients with hepatocellular carcinoma (HCC) for orthotopic liver transplantation (OLT) remains controversial. Since there is a trend to expand the transplant criteria for HCC patients, we reviewed the data of patients with HCC who had received OLT at our institute to determine their survival and prognostic factors., Methods: A total of 67 patients with HCC who had undergone OLT from April 2001 through December 2003 were reviewed retrospectively. Selection OLT candidates with HCC was dependent on the anatomical characteristics and/or the severity of underlying liver cirrhosis. The 67 patients were followed up for more than 6 months after transplantation. Their survival rate was calculated by the Kaplan-Meier method. Univariate and multivariate analyses using the Cox proportional hazards regression model were performed to reveal the factors affecting the survival rate., Results: No perioperative death occurred in this series. The 1- and 2-year cumulative survival rates were 90.0% and 65.6%, and the disease-free survival (DFS) rates were 77.5% and 62.5% respectively. Univariate analysis revealed the tumor size, portal vein tumor thrombus (PVTT), serum alpha-fetoprotein level, bilobular distribution of tumors, pTNM stage and histological differentiation were statistically significant factors affecting the DFS (P < 0.05). Multivariate analysis showed tumor size and PVTT were independent and statistically significant factors affecting the DFS (P = 0.005 and 0.010, respectively). In this series, all but 2 received systemic chemotherapy, among them 13 had tumor recurrence within 8 months after OLT., Conclusions: OLT is indicated for patients with HCC, even for some patients with end-stage liver disease who may survive longer without tumor recurrence. Adjuvant chemotherapy may decrease the recurrence of HCC after OLT.

Objective: To study the technique and effect of transcervical resection of myoma (TCRM) in treatment of hysteromyoma., Methods: 962 women suffering type 0 hysteromyoma (n = 281), 316 type 1 hysteromyoma (n = 316), type 2 hysteromyoma (n = 282), submucous and intramural myoma (n = 34), cervical myoma (n = 11), prolapse myoma (n = 23), and adenomyoma (n = 15) underwent TCRM with "five-step technique", monitored by B-ultrasound or laparoscopy, Follow-up lasted more than 6 months., Results: The primary operation successful rate was 99.77%. The mean size and depth of uterus were 7.44 +/- 1.3 gestation weeks and (8.31 +/- 1.43) cm, the diameter of the biggest myoma was 7.2 cm. The mean weight of the resected tissues was (22.63 +/- 31.41) g, and the mean operation time and blood loss during the operation were 32.50 +/- 172.72 minutes and (7.75 +/- 19.49) ml. No transfusion was needed. The complications included postoperative fever (3 cases), uterine bleeding (1 case), uterine perforation (1 case), and TURP syndrome (2 cases). Postoperative scanty menstrual rate was 100% in the type 0 hysteromyoma group, 99.1% in the type I hysteromyoma group, 94.02% in the type II hysteromyoma group, 100% in the cervical myoma and prolapse myoma group, 84% in the multiple myoma and intramural myoma group, and 87% in the adenomyoma group respectively. The alleviation rates of dysmenorrhea and anemia were 78% and 82.95% respectively. Those who have severe complications of internal medicine showed obvious improvement. The residual myoma of 2 cases were resected during the secondary operation 9 days and 3 months after the primary operation. 455 cases (52.17%) resumed their work in 1 month postoperatively. 32 living infants were delivered., Conclusion: Safe and highly effective, TCRM can be the first choice in treatment of submucous and intramural hysteromyoma.

Aim: Hepatocellular carcinoma(HCC) is a significant health problem in China. But the molecular mechanisms of HCC remains unclear. APC/beta-Catenin/Tcf signaling pathway, also known as Wnt pathway, plays a critical role in the development and oncogenesis. As little is known about the alteration of human T-cell transcription factor-4 (hTcf-4) gene in HCC, it is of interest to study the expression and mutation of hTcf-4 gene in HCC and the relationship between hTcf-4 gene and progression of HCC., Methods: Reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect the expression of hTcf-4 mRNA in 32 HCC and para-cancerous tissues and 5 normal liver tissues. PCR-single strand conformation polymorphism (PCR-SSCP) method was used to detect the mutation of hTcf-4 exons 1, 4, 9 and 15 in HCC. The correlation of expression and mutation of the hTcf-4 gene with clinicopathological characteristics of HCC was also analyzed., Results: RT-PCR showed that the expression rate of hTcf-4 mRNA in HCC, para-cancerous tissues and normal liver tissues was 90.6 %, 71.9 % and 80 %, respectively. The gene expression level in tumor was 0.71+/-0.13, much higher than that in para-cancerous liver 0.29+/-0.05 and normal liver 0.26+/-0.05 (P<0.001), although there was no significant difference in gene expression level between para-cancerous tissues and normal liver (P>0.05). Furthermore, hTcf-4 gene expression was closely associated with tumor capsule status and intrahepatic metastasis of HCC. On SSCP, 2 of 32 cases of HCC (6.25 %) displayed characteristic mutational mobility shifts in exon 15 of the hTcf-4 gene. No abnormal shifting bands were observed in para-cancerous tissues., Conclusion: The high expression level of hTcf-4 in HCC, especially in tumors with metastasis, suggests that the over-expression of hTcf-4 gene may be closely associated with development and progression of HCC, but the mutation of this gene seemed to play less important role in this respect.