Objective To obtain short peptides which specifically binds to HepG2 cell line from 12 peptide libraries, and lay foundation for screening and identifying the new liver cancer markers for early diagnosis and treatment of liver cancer. Methods The liver cancer cell line HepG2 was used as the antigen and LO-2 as the absorber cells for subtraction biopanning from a phage display peptide library at 37℃. The positive phage clones were identified by cell enzyme-linked immunosorbentassay (EL1SA), and the identity of DNA sequence and amino acids were analyzed. Results After 3 rounds of screening, 30 phage clones were identified by EL1SA, ZS-9 of them bind to the HepG2 specifically. The amino acid sequence was blast in NCBI and Swiss-Prot, the results show that the sequence has not identity with the known genes and proteins in the database, and the sequence was not reported in literature. All these show that we had discovered several novel liver cancer associated antigen ligand. Conclusion Several candidate peptide binding to liver cancer specifically have been selected by phage display technology, providing the potential uses for early diagnosis of liver cancer or targeted therapy for liver cancer.

Objective To investigate the effects of rhGH on hypoalbuminemia in cirrhotic rats after partial hepatectomy. Methods Thirty rats were randomly divided into normal control group ( n =6), liver cirrhosis group (LC group, n =6), liver cirrhosis and hepatectomy group (LCH group, n =6), PN (parenteral nutrition) group ( n =6, given PN after hepatectomy) and rhGH+PN group ( n =6,given rhGH and PN after hepatectomy). Liver function and blood glucose were measured. The expression of ALB mRNA was detected by RT PCR. Liver Ki67 immunohistochemistry was studied. Results Compared with PN group, serum ALP was lower; serum ALB and blood glucose were higher in rhGH+PN group. The expression of hepatic ALB mRNA was higher, and hepatic Ki67 labeling index was higher as well in this group. Conclusion rhGH can improve hypoalbuminemia after partial hepatectomy in cirrhotic rats with partial hepatectomy.

Objective To study the treatment of hepatocellular carcinoma(HCC) complicated with bile duct thrombus.Methods Restrospective study was performed on the treatment of 34 cases of HCC complicated with bile duct tumor thrombus from January 1995 to December 2002 admitted to our hospital.Results Among the 34 cases, one patient had no treatment,6 cases received PTCD and 27 cases received operation.In surgical treatment patients,the postoperative 30 days mortality was 22.2%(6/27),the morbidity was 55.6%(15/27).In 16 patients who received hepatectomy with thrombectomy and biliary duct drainage, the survival time was 1～27 months with a median survival time of 16.5 months.Conclusion The prognosis of HCC complicated with bile duct tumor thrombus is poor. Early diagnosis and aggressive operative are the key points to improving the survival time of patients.

ObjectiveTo explore the effects of different nutrition methods on the number and activity of natural killer (NK) cells in liver after hepatectomy in cirrhotic rats. MethodsTwenty thioacetamide-induced cirrhotic rats were randomly divided into enteral nutrition (EN) group ( n =10) and parenteral nutrition (PN) group ( n =10), in which EN or PN was administered for 5 consecutive days 1 day after hepatectomy. The percentage of the NK cells in livers was examined by flow cytometry. Activity of NK cells was measured by 4 h51Cr-release assay.ResultsCompared with the PN group, the percentage (22.2 ％ ± 1.7％ vs. 17.4％ ± 1.8％, P =0.000) and the activity of NK cells (41.6％ ± 2.3％ vs. 37.1％ ± 2.2％, P =0.000) in the livers were significantly higher in the EN group. ConclusionEN after posthepatectomy can ameliorate the percentage and the activity of NK cells in the livers in cirrhotic rats.

Objective To investigate the role of p21-activated kinase 1 (PAK1) in colorectal mucosal carcinogenesis and the relationship between PAK1 expression and biological behavior of colorectal carcinoma. Method PAK1 was detected with immunohistochemical method in 10 normal colorectal mucosas,40 colorectal villous or tubular adenomas and 60 colorectal carcinomas. Results The positive rate for PAK1 was 10.0% in normal colorectal mucesas,and 25.0%, 33.3% and 33.3% in slight, moderate and severe dys-plastic adenomas, respectively, 65.0% was found in colorectul carcinomas. The positive rate for PAK1 in col-orectal carcinomas was higher than that in normal colorectal mucosas(P<0.01), colorectal villous or tubular adenomas(P<0.01), slight dysplastic adenomas(P<0.01) and moderate dysplastic adenomas (P<0.05).The positive rate for PAK1 of poor differentiated colorectal carcinomas was higher than that of high differentiated ones (P<0.05), and the positive rate for PAK1 of patients with lymph node metastasis was higher than that of patients without lymph node metastasis (P<0.05). In clinical stages, the positive rate for PAK1 in Dukes C and D stages patients was higher than that in Dukes A stage patients, respectively (P<0.05). Conclusion PAK1 maybe play some role in the process of carcinogenesis of colorectal mucesa, and be used as an useful marker for assessment of the biological behavior and prognosis of colorectal carcinoma.

STAT-3 and Ki67 protein were significantly higher(P＜0.05)in the EN 5 days group.Conclusion Post-hepatectomy EN in cirrhotic rats can speed up liver function recovery,prevent cholestasis,and promote liver protein synthesis and liver regeneration.

Objective To find the liver cancer specific peptide for serological screen of liver cancer patients via screening phage-display peptide library. Methods Fifteen sera from liver cancer patients and physical examinates were collected for the four-round screening with Ph. D. 12TM phage display peptide kit. Highly specific phage monoclones were selected based on the ELISA results of the serological assay. The peptide labeled with FITC was synthesized according to the DNA sequencing of the optimal monoclone and tested with serum via fluorescent imagery. Results Nine highly specific monoclones were found among 50 selected ones after 4 rounds of screenings. The positive rate of the optimal monoclone,ZH-3, reached 46.7 %. The peptide sequence of ZH-3 was concluded by DNA sequencing as SAHGTSTGVPWP. Desirable specificity and affinity were also shown in the serum of liver cancer patients. Conclusion The peptide ZH-3 can be used as a diagnostic reagent for liver cancer.

AIM To investigate the effects of pioglitazone on cardiac hypertrophy in vitro. METHODS Hypertrophy in neonatal rat cardiac myocytes (MC) and cardiac nonmyocytes (NMC) was established with angiotensinⅡ (AngⅡ). mRNA expression of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) was measured by reverse transcription polymerase chain reaction (RT PCR). MTT assay and 3H TdR uptake was used to estimate proliferation of NMC. The surface area of MC was analyzed by the aid of NIH Image J software, and the synthetic rate of protein in MC was detected by 3H leucine incorporation. RESULTS In the condition of hypertrophy, increases of surface area,mRNA expression of ANP and BNP and 3H leucine incorporation in MC and an increase of proliferation in NMC were detected, but no changes in mRNA expression of ANP and BNP in NMC. Pioglitazone inhibited the changes above and reduced mRNA expression of ANP and BNP in NMC in a dose dependent manner. CONCLUSION The results demonstrate that pioglitazone inhibits cardiac hypertrophy in vitro and it suggests that pioglitazone has a potential role in the prevention and treatment of cardiac diseases such as cardiac hypertrophy.

Objective To evaluate the effects of branched-chain amino acids-enriched early parenteral and enteral nutrition on the liver function and serum aminograms in cirrhotic rats after partial hepatectomy. Methods In this prospective randomized controlled study, 24 cirrhotic rats, induced by thioacetamide, were randomized into three groups: enteral nutrition (EN) group, EN + branched-chain amino acid (BCAA) group, and parenteral nutrition (PN) + BCAA group. After receiving partial hepatectomy, rats in all three groups were nutritionally supported with equal amount of calorie and nitrogen contents from the 1st postoperative day ( PO day 1 ) to PO day 5. On PO day 6, parameters including body weight, liver functions, prealbumin, transferring, and serum aminograms were measured or determined, and the level of liver albumin mRNA was detected by reversal transcription-polymerase chain reaction and morphological examinations such as HE staining and immunohistochemical staining, which were assessed by index of Ki67 protein index. Results Body weight was significantly decreased in all three groups on PO day 6 (P ＜0.05 ). Compared with EN + BCAA group, serum aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase after partial hepatectomy were significantly higher in PN + BCAA group (P ＜0.05 ). Serum alkaline phosphatase level was significantly higher in PN + BCAA group than in EN group ( P ＜0. 05). The level of prealbumin was significantly lower in PN + BCAA group when compared with EN group or EN +BCAA group ( both P ＜ 0. 05 ), although no such significant difference was noted in terms of transferrin ( P ＞0. 05 ). The levels of leucine and isoleucine elevated while those of tyrosine, phenylalanine, arginine and tryptophan declined in PN + BCAA group or EN + BCAA group when compared with EN group ( P ＜ 0. 05 ). Aminograms were not significantly different between EN + BCAA group and PN + BCAA group ( P ＞ 0. 05 ). Levels of total amino acid and aromatic amino acid (AAA) were significantly lower while BCAA and ratio between BCAA and AAA (BCAA/AAA) were significantly higher in PN + BCAA group or EN + BCAA group than in EN group (P ＜ 0. 05 ).Significantly lower level of albumin mRNA and index of Ki67 were observed in PN + BCAA group than in EN group or EN + BCAA group (P ＜ 0.05 ) on PO day 6. Conclusions BCAA-enriched EN or PN reverses amino acid disequilibrium and restores BCAA/AAA in cirrhotic rats after partial hepatectomy. Compared with PN, EN is superior in improving postoperative liver function, promoting protein synthesis, and speed up tissue regeneration in the postoperative liver. However, it still can not restore serum albumin in a short term.

Objective To observe the levels of serum VEGF in different molecular subtypes of breast cancer and explore its relationship with response to neoadjuvant chemotherapy. Methods Levels of serum VEGF in 110 cases with breast cancer underwent neoadjuvant chemotherapy were detected by ELISA prior to and after 3 cycles of neoadjuvant chemotherapy. Clinical response to neoadjuvant chemotherapy was evaluated by physical examination and ultrasonography. Results Levels of serum VEGF were significantly increased in breast cancer patients with≥4 lymph node metastasis than those with < 4 lymph node[(307.31 ± 101.42) pg/mL vs. (170.16 ± 73.07) pg/mL,P = 0.017]. Patients with positive HER-2 status had significantly higher levels of serum VEGF than those with HER-2 negative status [(235.15 ± 88.42 ) pg/mL vs. (179.82 ± 69.90) pg/mL, P = 0.024]. No significant difference was observed among age , menopausal status and hormone status. In patients with neoadjuvant chemotherapy of cCR and Cpr,the mean levels of serum VEGF were (205.75 ± 78.12) pg/mL and (226.04 ± 89.04) pg/mL, respectively. After 3 cycles of chemotherapy, levels of serum VEGF decreased to (145.15 ± 67.08) pg/mL and (161.27 ± 93.57) pg/mL. There was significant difference between two groups (P=0.009,0.014). In patients with SD or PD response, no significant difference was observed between levels of serum VEGF before and after chemotherapy (P = 0.577). Conclusions Levels of serum VEGF in breast cancer correlate with lymph nodes metastasis and HER-2 status and may decrease after neoadjuvant chemotherapy. However,whether or not the levels of serum VEGF can be used as a biomarker for response to neoadjuvant chemotherapy needs more further studies.