Effect of postoperative adjuvant chemotherapy on prognosis of patients with intrahepatic cholangiocarcinoma:a multicenter retrospective study / 中华外科杂志

Objectives: To examine the influence of adjuvant chemotherapy after radical resection on the survival of patients with intrahepatic cholangiocarcinoma(ICC) and to identify patients who may benefit from it. Methods: The clinical and pathological data of 654 patients with ICC diagnosed by postoperative pathology from December 2011 to December 2017 at 13 hospitals in China were collected retrospectively. According to the inclusion and exclusion criteria,455 patients were included in this study,including 69 patients (15.2%) who received adjuvant chemotherapy and 386 patients (84.8%) who did not receive adjuvant chemotherapy. There were 278 males and 177 females,with age of 59 (16) years (M(IQR))(range:23 to 88 years). Propensity score matching (PSM) method was used to balance the difference between adjuvant chemotherapy group and non-adjuvant chemotherapy group. Kaplan-Meier method was used to plot the survival curve,the Log-rank test was used to compare the difference of overall survival(OS) and recurrence free survival(RFS)between the two groups. Univariate analysis was used to determine prognostic factors for OS. Multivariate Cox proportional hazards models were then performed for prognostic factors with P<0.10 to identify potential independent risk factors. The study population were stratified by included study variables and the AJCC staging system,and a subgroup analysis was performed using the Kaplan-Meier method to explore the potential benefit subgroup population of adjuvant chemotherapy. Results: After 1∶1 PSM matching,69 patients were obtained in each group. There was no significant difference in baseline data between the two groups (all P>0.05). After PSM,Cox multivariate analysis showed that lymph node metastasis (HR=3.06,95%CI:1.52 to 6.16,P=0.039),width of resection margin (HR=0.56,95%CI:0.32 to 0.99,P=0.044) and adjuvant chemotherapy (HR=0.51,95%CI:0.29 to 0.91,P=0.022) were independent prognostic factors for OS. Kaplan-Meier analysis showed that the median OS time of adjuvant chemotherapy group was significantly longer than that of non-adjuvant chemotherapy group (P<0.05). There was no significant difference in RFS time between the adjuvant chemotherapy group and the non-adjuvant chemotherapy group (P>0.05). Subgroup analysis showed that,the OS of female patients,without HBV infection,carcinoembryonic antigen<9.6 μg/L,CA19-9≥200 U/ml,intraoperative bleeding<400 ml,tumor diameter>5 cm,microvascular invasion negative,without lymph node metastasis,and AJCC stage Ⅲ patients could benefit from adjuvant chemotherapy (all P<0.05). Conclusion: Adjuvant chemotherapy can prolong the OS of patients with ICC after radical resection,and patients with tumor diameter>5 cm,without lymph node metastasis,AJCC stage Ⅲ,and microvascular invasion negative are more likely to benefit from adjuvant chemotherapy.

Comparison of follow-up treatment regimens for colorectal cancer liver metastases without objective response to neoadjuvant chemotherapy: direct surgery or surgery after second-line chemotherapy / 中华外科杂志

Objective: To compare the effect of direct surgery or surgery after second-line chemotherapy for colorectal cancer patients with liver metastases who did not achieve objective remission after neoadjuvant chemotherapy. Methods: A retrospective case cohort study was used. The clinical and pathological data of 107 patients with colorectal cancer liver metastases who did not achieve objective response to neoadjuvant chemotherapy at Department of Hepatobiliary Surgery,Cancer Hospital,Chinese Academy of Medical Sciences from December 2008 to December 2016 were retrospectively collected. There were 71 males and 36 females, median age was 57 years (range: 28 to 79 years). According to the different treatment regimens after neoadjuvant chemotherapy,107 cases were divided into a direct surgery group (direct group,n=65) and an operation after receiving second-line chemotherapy group (second-line group,n=42). The propensity score matching(PSM) of the Logistic regression model was used to match the bilobar distribution of liver metastases and the number of first-line chemotherapy cycles in the two groups of patients. The caliper value was set to 0.10 and the matching ratio was 1∶2. T test, Mann-Whitney U test, χ2 test or Fisher's exat test was used to analyzed the data between the tuo groups, respectively. Survival analysis design was used to investigate the difference in prognosis between the two groups of patients. Results: The follow-up time(M(IQR)) was 56.3(34.3) months (range: 2.1 to 95.0 months),and all patients were followed up. After PSM,there were 28 cases in the direct group and 42 cases in the second-line group, there were no significant differences in whether R0 resection was feasible,blood loss,blood transfusion,postoperative complications and postoperative hospital stay between the two groups (all P>0.05). The 1,3,and 5-year progression-free survival(PFS) rates of the direct group were 40.0%,16.5%,and 11.0%,and the 1,3,and 5-year overall survival(OS) rates were 98.5%,61.2%,and 41.4%,respectively, the second-line group 1,3,5 years PFS rates were 35.7%,14.3%,14.3%,1,3,5-year OS rate were 95.2%,55.1%,44.4%,respectively. The median PFS time of the direct group and the second-line group was 8.5 months and 7.5 months,respectively,and the difference was not statistically significant (P=0.826). The median OS time of the direct group and the second-line group were 33.8 months and 46.9 months,respectively. The difference was not statistically significant(P=0.646).The median PFS time of the direct group and second-line chemotherapy complete remission and partial remission group(CR/PR group) was 10.2 months and 9.1 months,respectively,and the difference was not statistically significant(P=0.669). The median OS time of the direct group and the second-line CR/PR group was 51.0 months and 46.9 months,respectively,and the difference was not statistically significant(P=0.427). The results of survival analysis suggested that major liver resection was an independent prognosis factor for PFS (HR=1.809,95%CI: 1.067 to 3.067,P=0.028) and OS(HR=2.751,95%CI: 1.317 to 5.747,P=0.007). Second-line chemotherapy was not an independent prognostic factor for PFS (HR=0.945, 95%CI:0.570 to 1.567,P=0.828) and OS (HR=0.866,95%CI: 0.468 to 1.602,P=0.646). Conclusions: There is no significant difference in the short-term outcome and long-term prognosis between direct surgery patients and second-line chemotherapy followed by surgery. Second-line chemotherapy is not an independent prognostic factor for colorectal cancer liver metastases patients who fail to achieve objective response after neoadjuvant chemotherapy.

Stereotactic body radiation therapy for patients with lung and liver oligometastases from colorectal cancer: a phase Ⅱ trial / 中华肿瘤杂志

Objective: To explore the safety and effectiveness of stereotactic body radiation therapy (SBRT) for oligometastases from colorectal cancer (CRC). Methods: This is a prospective, single-arm phase Ⅱ trial. Patients who had histologically proven CRC, 1 to 5 detectable liver or lung metastatic lesions with maximum diameter of any metastases ≤5 cm were eligible. SBRT was delivered to all lesions. The primary endpoint was 3-year local control (LC). The secondary endpoints were treatment-related acute toxicities of grade 3 and above, 1-year and 3-year overall survival (OS) and progression free survival (PFS). Survival analysis was performed using the Kaplan-Meier method and Log rank test. Results: Petients from 2016 to 2019 who were treated in Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College. Forty-eight patients with 60 lesions were enrolled, including 37 liver lesions and 23 lung lesions. Forty-six patients had 1 or 2 lesions, with median diameter of 1.3 cm, the median biologically effective dose (BED(10)) was 100.0 Gy. The median follow-up was 19.5 months for all lesions. Twenty-five lesions developed local failure, the median local progression free survival was 15 months. The 1-year LC, OS and PFS was 70.2% (95% CI, 63.7%~76.7%), 89.0% (95% CI, 84.3%~93.7%) and 40.4% (95%CI, 33.0%~47.8%). The univariate analysis revealed that planning target volume (PTV) and total dose were independent prognostic factors of LC (P<0.05). For liver and lung lesions, the 1-year LC, OS and PFS was 58.7% and 89.4% (P=0.015), 89.3% and 86.5% (P=0.732), 30.5% and 65.6% (P=0.024), respectively. No patients developed acute toxicity of grade 3 and above. Conclusion: SBRT is safe and effective treatment method for oligometastases from CRC under precise respiratory motion management and robust quality assurance.

Prognostic Analysis of 102 Patients with Synchronous Colorectal Cancer and Liver Metastases Treated with Simultaneous Resection / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The liver is the most common site for colorectal cancer (CRC) metastases. Their removal is a critical and challenging aspect of CRC treatment. We investigated the prognosis and risk factors of patients with CRC and liver metastases (CRCLM) who underwent simultaneous resections for both lesions.</p><p><b>METHODS</b>From January 2009 to August 2016, 102 patients with CRCLM received simultaneous resections of CRCLM at our hospital. We retrospectively analyzed their clinical data and analyzed their outcomes. Overall survival (OS) and disease-free survival (DFS) were examined by Kaplan-Meier and log-rank methods.</p><p><b>RESULTS</b>Median follow-up time was 22.7 months; no perioperative death or serious complications were observed. Median OS was 55.5 months; postoperative OS rates were 1-year: 93.8%, 3-year: 60.7%, and 5-year: 46.4%. Median DFS was 9.0 months; postoperative DFS rates were 1-year: 43.1%, 3-year: 23.0%, and 5-year 21.1%. Independent risk factors found in multivariate analysis included carcinoembryonic antigen ≥100 ng/ml, no adjuvant chemotherapy, tumor thrombus in liver metastases, and bilobar liver metastases for OS; age ≥60 years, no adjuvant chemotherapy, multiple metastases, and largest diameter ≥3 cm for DFS.</p><p><b>CONCLUSIONS</b>Simultaneous surgical resection is a safe and effective treatment for patients with synchronous CRCLM. The main prognostic factors are pathological characteristics of liver metastases and whether standard adjuvant chemotherapy is performed.</p>

Excision Repair Cross-complementation Group 1 is a Prognostic Biomarker in Patients with Colorectal Cancer Receiving Chemotherapy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Conflicting results about the association between expression level of excision repair cross-complementation group 1 (ERCC1) and clinical outcome in patients with colorectal cancer (CRC) receiving chemotherapy have been reported. Thus, we searched the available articles and performed the meta-analysis to elucidate the prognostic role of ERCC1 expression in patients with CRC.</p><p><b>METHODS</b>A thorough literature search using PubMed (Medline), Embase, Cochrane Library, Web of Science databases, and Chinese Science Citation Database was conducted to obtain the relevant studies. Pooled hazard ratios (HR s) or odds ratios (OR s) with 95% confidence intervals (CI s) were calculated to estimate the results.</p><p><b>RESULTS</b>A total of 11 studies were finally enrolled in this meta-analysis. Compared with patients with lower ERCC1 expression, patients with higher ERCC1 expression tended to have unfavorable overall survival (OS) (HR = 2.325, 95% CI: 1.720-3.143, P < 0.001), progression-free survival (PFS) (HR = 1.917, 95% CI: 1.366-2.691, P < 0.001) and poor response to chemotherapy (OR = 0.491, 95% CI: 0.243-0.990, P = 0.047). Subgroup analyses by treatment setting, ethnicity, HR extraction, detection methods, survival analysis, and study design demonstrated that our results were robust.</p><p><b>CONCLUSIONS</b>ERCC1 expression may be taken as an effective prognostic factor predicting the response to chemotherapy, OS, and PFS. Further studies with better study design and longer follow-up are warranted in order to gain a deeper understanding of ERCC1's prognostic value.</p>

Evaluation of eight different clinical staging systems associated with overall survival of chinese patients with hepatocellular carcinoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Hepatocellular carcinoma (HCC) is a common cancer in China, an area of high hepatitis B virus (HBV) infection. Although several staging systems are available, there is no consensus on the best classification to use because multiple factors, such as etiology, clinical treatment and populations could affect the survival of HCC patients.</p><p><b>METHODS</b>This study analyzed 743 HBV-related Chinese HCC patients who received surgery first and evaluated the predictive values of eight different commonly used staging systems in the clinic.</p><p><b>RESULTS</b>The overall 1-, 3-, 5-year survival rates and a median survival were 91.5%, 70.3%, 55.3% and 72 months respectively. Barcelona Clinic Liver Cancer (BCLC) staging systems had the best stratification ability and showed the lowest Akaike information criterion (AIC) values (2896.577), followed by tumor-node-metastasis 7 th (TNM 7 th ) (AIC = 2899.980), TNM 6 th (AIC = 2902.17), Japan integrated staging score (AIC = 2918.085), Tokyo (AIC = 2938.822), Cancer of the Liver Italian Program score (AIC = 2941.950), Chinese University Prognostic Index grade (AIC = 2962.027), and Okuda (AIC = 2979.389).</p><p><b>CONCLUSIONS</b>BCLC staging system is a better staging model for HBV infection patients with HCC in Chinese population among the eight currently used staging systems. These identifications afford a large group of Chinese HCC patients with HBV infection and could be helpful to design a new staging system for a certain population.</p>

Prognosis of hepatocellular carcinoma: a study of 832 cases / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the prognostic factors of hepatocellular carcinoma.</p><p><b>METHODS</b>The purpose of this study was to retrospectively analyze the surgical outcomes of hepatocellular carcinoma (HCC) in 832 patients who underwent hepatic resection between February 2002 and June 2010 in the Cancer Hospital of Chinese Academy of Medical Sciences. Post-resection prognostic factors were assessed using a univariate Kaplan-Meier analysis and a multivariate Cox proportional hazards model.</p><p><b>RESULTS</b>The overall 1-, 3- and 5-year survival rates were 92.0%, 70.2% and 53.6%, respectively. The disease free survival rates (DFS) were 90.2%, 61.5% and 40.5%, respectively. The univariate analysis showed that a better prognosis for overall survival (OS) was associated with asymptomatic presentation, small tumor, single lesion, high-grade histological differentiation, no vascular tumor embolus, negative serum alpha-fetoprotein (AFP), negative serum alkaline phosphatase (ALP), Child-Pugh class A, no ascites, no/mild cirrhosis, new surgical techniques, no blood transfusion, no regional lymph node metastasis, no major vascular invasion, and no extra-hepatic invasion. The multivariate analysis showed that asymptomatic presentation, small tumor, single lesion, no tumor embolus, negative serum alpha-fetoprotein (AFP), no regional lymph node metastasis, no major vascular invasion, no extra-hepatic invasion, no/mild cirrhosis, and surgical techniques are independent factors for a longer overall survival.</p><p><b>CONCLUSIONS</b>The prognosis of HCC after resection is influenced by a number of factors. Therefore, regularly screening and early diagnosis, applying surgical techniques to minimize the liver injury, and preventing the aggravation of cirrhosis are important measures to improve the overall survival of HCC patients. For those patients with high risk factors of recurrence, routine follow-up is one of the best methods to be recommended.</p>

Clinicopathological and prognostic analysis of primary clear cell carcinoma of the liver / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathological features and prognostic factors of primary clear cell carcinoma of the liver (PCCCL).</p><p><b>METHODS</b>The clinical data of 41 PCCCL patients who underwent hepatic resection for PCCCL from October 1998 to June 2012 in our department were retrospectively analyzed. There were 31 male and 10 female patients. The median age was 56 years (range, 25 to 80 years), and the diagnosis was confirmed by postoperative pathological examination. The data of 106 well or moderately differentiated non-clear cell hepatocellular carcinoma (HCC) patients and 86 poorly differentiated non-clear cell HCC patients who underwent hepatic resection in the same period in our hospital in the same period were compared. The χ(2) test or Fischer's exact test, as appropriate, was used to compare group frequencies. Survival analysis was estimated by Kaplan-Meier method. Cox proportional hazards model was used in multivariate analysis.</p><p><b>RESULTS</b>The proportion of fibrous capsule formation in the PCCCL tumors (46%, 19/41) was significantly higher than that of the other two groups (P < 0.05), whereas the PCCCL group had a lower rate of intravascular tumor embolus (2/41) and vascular invasion (1/41) (P < 0.05). The median survival time of PCCCL group was 65 months, the 1-, 3-, 5-year survival rates for PCCCL patients were 90.2%, 67.4% and 42.0%, significantly better than that of poor differentiated NCCHCC group's (82.9%, 33.3%, 7.2%, P < 0.01). However, there were no statistic significant differences between PCCCL group and well or moderately differentiated NCCHCC group (84.7%, 55.7%, 34.4%, P > 0.05). Tumor capsule formation was an independent favorable prognostic factor. In contrast, preoperative serum α-fetoprotein (AFP) level and hepatitis B virus infection were independent unfavorable prognostic factors for PCCCL.</p><p><b>CONCLUSIONS</b>PCCCL is a rare, low degree malignant pathological subtype of HCC. Surgical resection may achieve favorable prognosis and even long-term survival.</p>

Surgical management of nodular hyperplasia tumor-like hepatic lesions: a study of 72 cases / 中华外科杂志

<p><b>OBJECTIVE</b>To explore and improve the diagnosis and treatment of benign hyperplasia tumor-like hepatic lesion.</p><p><b>METHODS</b>The clinical data of 72 patients who had undergone hepatic resection for benign non-cystic hepatic lesions between January 1987 and December 2010 were analyzed retrospectively. There were 46 male and 26 female patients. The median age was 49 years (ranging 15 to 72 years), and diagnosis were confirmed by postoperative pathological examination. Thirty-four cases had symptoms, such as abdominal discomfort in right upper quadrant, fever, fatigue. All the cases had undergone hepatic resection; totally 78 lesions were removed. The surgical procedure including hepatic lobectomy for 13 cases, hepatic segmentectomy for 19 cases and non-anatomy resection for 39 cases.</p><p><b>RESULTS</b>The finally diagnosis included focal nodular hyperplasia in 47 cases, adenomatous hyperplasia in 3 cases, hepatic dysplastic nodule in 3 cases, inflammatory pseudotumor in 3 cases, hepatic granuloma in 4 cases, nodular cirrhosis in 3 cases, hepatitis nodule in 6 cases, nodular regenerative hyperplasia in 1 case, lymphoid hyperplasia in 1 case. The postoperative complication rate was low (19.4%, 14/72). The follow-up period was 6 to 96 months. There was no mortality caused by lesion. One patient developed recurrence after 3 years.</p><p><b>CONCLUSIONS</b>It is suggested that symptomatic lesions, lesions when malignancy cannot be excluded, and lesions which have canceration tendency, just like adenomatous hyperplasia, dysplastic nodule, and nodular cirrhosis regenerative hyperplasia need surgical resection. Operation is not necessary for other nodular hyperplasia lesions if the diagnoses are identified.</p>

Peri-operative treatment for hepatocellular carcinoma patients complied with cirrhosis and hypersplenism / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the influence of combined hepatectomy with splenectomy on safety of operation for hepatocellular carcinoma patients complied with cirrhosis and hypersplenism and the best peri-operative treatment of these patients.</p><p><b>METHODS</b>Clinical data of 177 hepatocellular carcinoma patients complied with cirrhosis and hypersplenism admitted from January 1999 to December 2009 were analyzed retrospectively. Among which, 71 patients received concomitant splenectomy with hepatectomy (splenectomy group), 106 patients only receive a hepatectomy (non-splenectomy group). The safety of operation, complications, liver function and WBC and PLT counts were compared between the two groups.</p><p><b>RESULTS</b>There was no significant difference of general conditions, counts of WBC and PLT between the two groups before operation. The counts of PLT at 1, 10, 30 day after operation were (88.4 ± 23.6) × 10⁹/L, (345.3 ± 98.2) × 10⁹/L and (210.8 ± 92.2) × 10⁹/L respectively in splenectomy group, which were significantly higher than that of non-splenectomy group (P < 0.05). The operation time of splenectomy group was (216 ± 105) min, which was longer than that of non splenectomy group (P < 0.05), but the blood loss and transfusion rate had not significantly difference between the two groups. The complication rates of splenectomy group and non-splenectomy group were 11.3% and 6.6% respectively, there was no significant difference between the two groups.</p><p><b>CONCLUSION</b>Combined hepatectomy with splenectomy will be safe for hepatocellular carcinoma patients complied with cirrhosis and hypersplenism as if the operative indication and increase the ability of peri-operative treatment are strictly obeyed.</p>

Analysis of prognostic factors influencing the outcome of intrahepatic cholangiocarcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To analyze the clinicopathologic factors influencing the outcome of surgically treated intrahepatic cholangiocarcinoma (ICC) and to explore the proper treatment choice of ICC.</p><p><b>METHODS</b>The clinicopathological data of 43 surgically treated ICC patients in our hospital were retrospectively analyzed. Of the 43 patients, hepatic resection was performed in 40 patients, ethanol injection in 2, and laparoscopic exploration alone in 1. Kaplan-Meier method and Cox regression model were used for the analysis of factors influencing survival after operation.</p><p><b>RESULTS</b>The accumulative 1-, 3- and 5-year survival rates were 64.4%, 30.9%, 25.8% for the whole group, and 74.7%, 33.3%, 27.8% for the 40 patients with hepatic resection, respectively. Univariate analysis revealed that tumor size, carcinoembryonic antigen (CEA) level, lymph node involvement and TNM stage were factors significantly affecting the survival (P < 0.05). Cox multivariate analysis demonstrated that only tumor size and lymph node involvement were the independent factors significantly affecting the survival (P < 0.05).</p><p><b>CONCLUSION</b>Our results show that tumor size and lymph node involvement are independent factors affecting the survival. CEA level and TNM stage are important prognostic factors for surgical management. Radical resection is still the optimal treatment for patient with intrahepatic cholangiocarcinoma.</p>

Clinical application of anatomic method of separation in hepatectomy / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the effectiveness of anatomic method of separation in hepatectomy methods of decreasing postoperative complication and mortality for liver cancer patients.</p><p><b>METHODS</b>The clinical data of 398 patients with liver malignant tumors, admitted in our hospital during 2001 to 2007, were retrospectively analyzed. The anatomic method group (group A) included 243 contiguous patients of liver cancer who received hepatectomy by anatomical method of separation, while the traditional method group (group B) included 155 patients of liver cancer who received hepatectomy by traditional method of separation during the same period. Blood loss and transfusion during operation, postoperative liver function, complication and mortality of operation, and postoperative hospital stay were compared between these two groups.</p><p><b>RESULTS</b>In group A, the tumor diameters ranged (6.02 +/- 3.24) cm, the operative blood loss was (445 +/- 240) ml, and 52 patients (24.3%) underwent blood transfusion [range: (520 +/- 280) ml]. No mortality and intraabdominal hemorrhage, liver function failure, or other severe complications were noted. Only 12 patients (4.9%) suffered mild complications. The postoperative hospital stay was (13.4 +/- 4.9) days. In group B, the tumor diameters ranged (5.84 +/- 2.93 cm, the operative blood loss was (1200 +/- 320) ml, and 53 patients (34.2%) underwent transfusion [range: (1400 +/- 623) ml]. Five patients (3.23%) died within 30 days after operation. The total complication morbidity was 16.1% (25/155). Five patients suffered intraabdominal hemorrhage and 5 experienced liver function failure. Postoperative hospital stay was (18.9 +/- 10.3) days. The volume of blood loss and transfusion in group A were significantly less than in group B (P < 0.05). Mortality, severe complications, and total complication morbidity were significantly lower in group A than in group B (P < 0.05, P < 0.01). The postoperative stay was also significantly shorter in group A than in group B (P < 0.05).</p><p><b>CONCLUSION</b>Anatomical method of separation is an effective method of hepatectomy with relatively low complication and mortality.</p>

Establishment of a sandwich ELISA method for detection of vascular endothelial growth factor in serum samples of hepatocellular carcinoma patients / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To establish a sandwich ELISA method for detecting vascular endothelial growth factor (VEGF) in sera of population and the patients with hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Full length and two truncated human VEGF cDNA sequences were amplified from a commercial plasmid pBLAST49-hVEGF by PCR and inserted into the prokaryotic-expression plasmid pET-32a or pGEX-2T. Various VEGF proteins were expressed and purified from E. coli in His-Trx or GST fusion forms. The specific VEGF antibodies were elicited in experimental rabbits and mice by immunization of the full length VEGF fusion protein His-Trx-VEGF1-165. After purification of antibodies with chromatograph of Protein G, a sandwich ELISA technique was established. Serum VEGF levels were evaluated in 229 adults and 291 HCC patients.</p><p><b>RESULTS</b>SDS-PAGE displayed that the molecular weights of the expressed full length (His-Trx-VEGF1-165), N-terminal (His-Trx-VEGF1-100) and C-terminal (GST-VEGF100-165) human VEGF fusion proteins were about 38KD, 31KD, and 33KD, respectively. Western blots confirmed that the prepared antisera were able to recognize both prokaryoticly and eukaryoticly expressed recombinant VEGF proteins. Assays of serially diluted His-Trx-VEGF1-100 by the established sandwich ELISA method showed that the linear range of the standard curve was 0.625-320 ng/mL, with the squared correlation coefficient R2 = 0.991. Screening of a serum panel containing 291 serum samples of HCC patients and 229 health adults revealed that the average VEGF level in HCC patients was higher than that in healthy controls, with a statically significant difference.</p><p><b>CONCLUSION</b>The established sandwich ELISA reflects the level of serum VEGF and provide scientific basis for screening metastasis and recurrence of HCC using serum VEGF as an index.</p>

Establishment of a sandwich ELISA method for detection of matrix metalloproteinase 9 protein in sera samples and preliminary application for hepatocellular carcinoma patients / 中华检验医学杂志

Objective Expression of recombinant human matrix metalloproteinase 9 (MMP9) protein in E coli,To establish a sandwich ELISA method for detecting MMP9 in sera and analyses of the difference of serum MMP9 in health population and the patient with hepatocellular carcinoma (HCC). Methods A fragment MMP9 sequence from nt 721-1 156 was amplified with PCR from the synthesized cDNAs of human liver tissues and inserted into the prokaryotic-expression plasmid pQE30.HIS-MMP9 fusion protein was expressed and purified from E coli MIS cells.The specific antibodies were elicited in rabbits and guinea pigs by immunization of the purified HIS-MMP9.After purifing antibodies with CNBr-activated Sepharose 4B,a sandwich ELISA technique was established.The serum MMP9 proteins were evaluated in 227 health adults and 193 HCC patients.Results SDS-PAGE displayed that the molecular-weight of the expressed fusion protein was about 17 000.The prepared antisera were able to recognize both recombinant and endogenously expressed MMP9 from human neutrophil and HepG2 cells.It was found that the average level of MMP9 proteins in HCC patients was higher than that in health control,showing significantly statistic difference.Conclusions The established method could reflect the level of serum MMPg.The data in this study supply scientific basis of generating methodology for screening metastasis and reoccurrence of HCC, using serum MMP9 as index.