AFP from fetal serum and serum of patients with primary hepatic cancer were purified by affinity chromatography. Then, the lentil lectin-reactive and nonreactive variants of these purified glycoproteins were prepared by affinity chromatography with immobilized lectin. Glycopeptides and oligosaccharides were prepared from variants by protease treatment and hydrazinolysis, respectively, and subjected to carbohydrate and amino acid analysis. A small difference in the carbohydrate compositions of each variant was observed. Analysis of chemical compositions of AFP variants is useful for the differential diagnosis between benign liver diseases and primary hepatic cancer.

Aim To investigate the effect of selective cyclooxygenase-2 inhibitor, NS-398, on cell apoptosis in leukemia cell line K562 cells and its underlying mechanism. Methods Apoptotic cell percentage was examined by flow cytometry(FCM). The protein expression of Bcl-2 and Caspase-3 was detected by Western blot, and the activity of Caspase-3 was checked by FCM. Results NS-398 induced the cell apoptosis in K562 cells in a dose-dependent manner. After NS-398 treatment, the expression of Bcl-2 protein was downregulated, whereas the expression of Caspase-3 protein was upregulated. Moreover,the activity of Caspase-3 was increased in a dose-dependent way after NS-398 treatment. Conclusion Selective cyclooxygenase-2 inhibitor,NS-398,significantly induced apoptosis in K562 cells. The underlying mechanism might be related to the downregulation of Bcl-2 and the activation of Caspase-3.

Objective To study the prevention and treatment of intra- or post- operative complications of radiofrequency ablation (RFA) for hepatic malignancy. Methods A series of 353 patients with primary hepatic carcinoma or secondary hepatic malignancy verified by pathological examinations and clinical manifestations underwent 452 times of RFA between October 1999 and March 2003, including 344 cases of percutaneous RFA and 9 cases of RFA during open surgery. There were 198 cases of primary hepatic carcinoma previously unoperated, 81 cases of recurrent hepatic carcinoma after surgery, and 74 cases of secondary hepatic malignancy. All the patients were followed up to make out short- or long-term surgical complications. Results Complications happened in 12 cases: skin burn by the electrode (2 cases), pneumothorax (1 case), right hydrothorax (1 case), vena cava thrombus (1 case), right bile duct injury and stricture (1 case), gastric external fistula (1 case), bile leakage in the left lateral lobe of the liver (1 case), hepatic abscess (1 case), external fistula of the ascending colon resulted from hepatic abscess (1 case), internal fistula of the duodenum, liver and right thorax resulted from hepatic abscess (1 case), and intra-abdominal hemorrhage (1 case). The incidence of complication was 2 65% (12/452) and the complication-related mortality was 8 33% (1/12). Conclusions RFA is a kind of minimally invasive procedure. But RFA especially percutaneous RFA may involve some risks for patients with the tumor located at the porta hepatic or the surface of the liver, or adhered to hollow organs, especially those who had a history of abdominal operation and adhesions of hollow viscera to the liver. Some complications can be prevented and effectively treated in the event of proper selection of patients, careful performance during the surgery, intensive postoperative care, effective hemostasia and efficient infection prevention.

Objective To explore the clinical significance of expression of MKK4 in osteosarcoma,and to discuss the relationship between MKK4 and osteosarcoma occurrence,development and prognosis.Methods The expression of MKK4 protein was determined with immunohistochemical SP method in 30 cases of osteosarcoma.Results The MKK4 was obviously expressed in both nucleus and cytoplasm in osteosarcoma.The expression of MKK4 in the cytoplasm of osteosarcoma was positively correlated with clinical stage (r =0.72,P =0.000),the expression of MKK4 in the nucleus of osteosarcoma had no correlation with clinical stage (r =0.44,P =0.080).MKK4 expression in the cytoplasm was higher in transfer osteosarcoma than no transfer osteosarcoma(x2 =9.69,P ＜0.01),while no obvious difference in the nucleus of osteosarcoma(x2 =1.00,P =1.000).Conclusion The MKK4 is highly expressed in the nucleus and cytoplasm of osteosarcoma,suggests that MKK4 may be related with the carcinogenesis development of osteosarcoma.The expression of MKK4 in the cytoplasm of osteosarcoma is related to the malignant,metastasis and prognosis of osteosarcoma,may be used as a potential prognostic indicator of judgment osteosarcoma.

Objective To prepare the anti-hUART1 polyclonal antibody and investigate the subcellular localization of hURAT1 protein. Methods The full-length hURAT1 gene was obtained by RT-PCR and inserted into the fusion expression vector pEGFP-N3,the predicated antigen epitope was cloned into GST fusion protein expression plasmid pGEX-5X-1, transformed into E. coli BL21 cells for expressing recombinant GST-hURAT1 protein induced by IPTG. The purified hURAT1-GST fusion protein was employed to immunize rabbit for preparing the polyclonal antibody. The expression of hURAT1 was analyzed by western-blot and immunohistochemistry in human kidney. pEGFP-hURAT1 was transfected into the LLC-PK1 cell in order to observe the subcellular localization of the gene using confocal microscopy. Results Specific anti-hURAT1 rabbit polyclonal antibody was obtained, and both Western-blot and immunohistochemistry showed that hURAT1 was expressed in the human kidney brush border,localized in the apical membrane of the LLC-PK1 cell. Conclusions hURAT1 protein was a membrane protein located in renal proximal tubule, which could be detected in the apical membrane. The anti-hURAT1 polyclonal antibody could be used for studying the physiological function of hURAT1 and its pathology.

Activities of N-acetyl-?-glucosaminidase (NAG) in sera of normal subjects and patients with hepatocellular carcinoma (HCC) and other diseases were microquantitatively determined by spectrophotometry. The results showed that after the reaction for 4 h, activity of serum NAG in patients with HCC (397.10 ? 174.97 nmol/ml) was obviously higher than that in normal subjects (280.00 ? 63.83 nmol/ml). The elevation of serum NAG in HCC patients had no relationship with their serum a-fetoprotein. However, the increase of serum NAG activity in some patients with benign liver diseases was always accompanied by the abnormality in their liver functions. It is suggested that serum NAG has some referable values in the diagnosis of HCC.

Aim To study the protective effect of total flavones of rhododendra pharmacological preconditioning(TFR-PP)on myocardial ischemia and reperfusion injury and its mechanisms involved in myocardial inflammation in rats.Methods In model group the isolated perfused rat hearts set up by Langendorff system were subjected to 30 min ischemia followed by 40 min reperfusion. The hearts in TFR-PP groups were subjected to three cycles of 5 min perfusion with and without TFR before 30 min ischemia followed by 40 min reperfusion. In all groups the activities of lactate dehydrogenase (LDH), creatine phosphokinase (CK), and myeloperoxidase (MPO) in myocardium, the expressions of nuclear factor-kappa B (NF-?B), tumor necrosis factor-alpha(TNF-?), and intercellular adhersion molecule-1 (ICAM-1) were measured, and the myocardial pathomorphological changes were examined.Results TFR-PP(25、50、100 mg?L-1) could inhibit markedly the reductions of LDH and SOD activities in myocardium induced by ischemia and reperfusion injury.100 mg?L-1 TFR-PP also significantly improved the pathomorphological changes of injury. TFR-PP (25、50、100 mg?L-1) could inhibit the expressions of NF-?B, TNF-? and ICAM-1 to varying degrees.Conclusion TFR-PP has marked protective effect on ischemia and reperfusion injury in isolated rat heart via inhibiting the inflammation of myocardium.

Objective The purpose of the study was to investigate the correlation in incidence and severity between peripheral arterial disease(PAD) and coronary artery disease and stroke in Chinese people,in order to assess the essentiality of Ankle Brachial Index(ABI) examination for all the patients.Methods Patients hospitalized to Departments of Cardiology,Neurology and Vascular Diseases from March 2006 to December 2007 were enrolled.ABI was measured,and the disease history was collected.Ejection fraction(EF),coronary arteriography and ultrasound examination of the cervical artery were recorded to analyze the relationship between them and ABI.The prevalence of decreased ABI was calculated.Results In 132 patients hospitalized in the Department of Cardiology,the prevalence of ABI

CXCR4, a receptor for stromal cell-derived factor-1 (SDF-1, also know as CXCL12) and a member of the chemokine receptor family, has been shown to play an important role in the proliferation, migration, invasion and metastasis of breast cancer cells. CXCR4 expression is an early event in breast cancer development and may serve as a biomarker for early detection and diagnosis of breast cancer. Expression of CXCR4 has been demonstrated to correlate with metastasis and poor prognosis in breast cancer. Inhibition of CXCR4 expression may be a potential target of breast cancer treatment.

Objective To present the experience of repairing advanced scrotal hypospadias with the combined use of tubed transverse preputial island flap (Duckett) and inversed scrotal raphe flap urethroplasty. Methods 19 cases were operated on with repairing distal urethral tube by Duckett preputial island flap and scrotal raphe flap urethroplasty with suturing together the adjoining ends of the two tubes on an oblique plane.The age of the patients ranged from 2 to 12 years. The length of urethroplasty was 5～9 cm. Results All the cases were followed up for 6 months to 4 years. 17 cases recovered fully on one stage operation. 2 were complicated by urethral fistula. Conclusions The results are satisfactory.The use of a casing tube is reasonable and nontraumatic.