Effects of p53 on apoptosis and proliferation of hepatocellular cardnoma cells heated with transcatheter arterial chemoembolization

AIM:To evaluate the effects of p53 on apoptosis and proliferation of hepatocellular carcinoma (HCC) cells treated with transcatheter arterial chemoernbolization (TACE).METHODS: A total of 136 patients with HCC received TACE and other management before surgery were divided into TACE group and non-TACE group.TACE group included 79 patients who had 1-5 courses of TACE before surgery,of them,11 patients had 1-4 courses of chemotherapy (group A),33 patients had 1-5 courses of chemotherapy combined with iodized oil (group B), 23 patients had 1-3 courses of chemotherapy,iodized oil and gelatin sponge (group C),12 patients had 1-3 courses of chemotherapy combined with iodized oil,ethanol and gelatin sponge (group D).Non-TACE group included the remaining 57 patients who had surgery only.The extent of apoptosis was analyzed by transferase mediated dUTP nick end labeling (TUNEL) staining.The expressions of p53, Bcl-2,Bax,Ki-67 and PCNA protein were detected by immunohistochemical method.RESULTS: P53 protein expressions in trabecular and clear cells in HCC specimens were significantly lower than that in pseudoglandar, solid, poorly differentiated or undifferentiated and sclerosis HCC (P<0.05) Expression of p53 protein in HCC cells increased with the increase of pathological grades(P<0.05),and correlated positively with expressions of Ki-67 and PCNA protein,and negatively with Bcl-2 to Bax protein expression rate and AI (P<0.05).Expression of p53 protein was significantly higher in group A than in groups B, C, D and the non-TACE group, and was higher in group B than in groups C and D,and lower in group D than in the non-TACE group (P<0.05).CONCLUSION:Expression of p53 protein can enhance proliferation of HCC cells and suppress apoptosis of HCC cells after TACE.

Clinical and experimental study on therapeutic effect of umbilical cord blood transplantation on severe viral hepatitis

AIM: To investigate the therapeutic effect of umbilical cordblood transplantation (UCBT) on patients with severe viralhepatitis and on liver lesions in rats.METHODS: One hundred and fifty three patients with severeviral hepatitis were included in the study between April 1990and July 2002. The patients were treated with adult plasmatransfusion (control), UCBT, plasma exchange (PE) and UCBTcombined with PE (UCBT+PE) respectively. The therapeuticeffectiveness was evaluated by serial determinations of liverfunction, lipids and immune function in all patients beforeand after the treatment. The model of experimental hepaticfailure was constructed in SD rats by injecting carbontetrachloride. Then, the rats were given normal saline, adultplasma or neonate cord blood intraperitoneally. After detectionof liver function, the rats were killed and morphologicalchanges of the liver were microscopically observed.RESULTS: UCBT group and UCBT+PE group had muchbetter improvement in liver and immune functions thancontrol group and PE group. The patients in UCBT+PE grouphad the best clinical efficacy. UCBT was safe and had noside effects. The animal experiment showed significantimprovements in liver function and survival rate in neonatecord blood group as compared with adult plasma group.The histopathology of rat′s liver indicated that neonate cordblood application could decrease the liver injury and increasehepatocellular regeneration.CONCLUSION: UCBT demonstrated a good therapeuticeffect on severe viral hepatitis and no obvious side effects.Umbilical cord blood can attenuate the liver lesions andreproduce hepatocyte. The treatment of UCBT combinedwith PE was much better than that of single plasma exchange, thus UCBT can enhance the therapeutic effect of plasma exchange on severe viral hepatitis.

Proteomics to display tissue repair opposing injury response to LPS-induced liver injury

AIM: To examine the protein expression alterations in liver injury/repair network regulation as a response to gut-derived lipopolysaccharide (LPS) treatment, in order to anticipate the possible signal molecules or biomarkers in signaling LPS-related liver injury.METHODS: Male BALB/c mice were treated with intraperitoneal (i.p.) LPS (4 mg/kg) and sacrificed at 0, 6, 24 and 30 h to obtain livers. The livers were stained with hematoxylin and eosin for histopathologic analyses. Total liver protein was separated by two-dimensional gel electrophoresis (2-DE). The peptide mass of liver injury or repair related proteins were drawn up and the protein database was searched to identify the proteins.RESULTS: Observations were as follows: (1) TRAIL-R2 was down regulated in livers of LPS-treated mice. TNFAIP1 was significantly up regulated at 6 h, then down- regulated at 24, 30 h with silent expression during senescent stage.(2) The amount of metaxin 2 and mitochondria import inner membrane translocase subunit TIM8a (TIMM8A) was increased upon treatment with LPS, (3) P34 cdc2 kinase was significantly up-regulated 30 h after LPS administration with silent expression during senescent, 6, 24 h treated stage. (4) The amount of proteasome activator 28 alpha subunit (PA28), magnesium dependent protein phosphatase(MDPP) and lysophospholipase 2 was decreased 6 h after LPS treatment but recovered or up-regulated 24 and 30 h after LPS treatment.CONCLUSION: LPS-treated mouse liver displaying a timedependent liver injury can result in expression change of some liver injury or repair related proteins.

Six-year follow-up of pancreatic β cell function in adults with latent autoimmune diabetes

AIM: To investigate the characteristics of the progression of islet β cell function in Chinese latent autoimmune diabetes in adult (LADA) patients with glutamic acid decarboxylase antibody (GAD-Ab) positivity, and to explore the prognostic factors for β cell function. METHODS: Forty-five LADA patients with GAD-Ab positivity screened from phenotypic type 2 diabetic (T2DM) patients and 45 T2DM patients without GAD-Ab matched as controls were followed-up every 6 mo. Sixteen patients in LADA1 and T2DM1 groups respectively have been followed-up for 6 years, while 29 patients in LADA2 and T2DM2 groups respectively for only 1.5 years. GAD-Ab was determined by radioligand assay, and C-peptides (CP) by radioimmune assay.RESULTS: The percentage of patients whose fasting CP(FCP) decreased more than 50% compared with thebaseline reached to 25.0% at 1.5th year in LADA1 group, and FCP level decreased (395.8±71.5 vs 572.8±72.3 pmol/L, P＜0.05) at 2.5th year and continuously went down to the end of follow-up. No significant changes of the above parameters were found in T2DM1 group. The average decreased percentages of FCP per year in LADA and T2DM patients were 15.8% (4.0-91.0%) and 5.2% (-3.5 to 35.5%, P= 0.000) respectively. The index of GAD-Ab was negatively correlated with the FCP in LADA patients (rs= -0.483, P = 0.000). The decreased percentage of FCP per year in LADA patients were correlated with GAD-Ab index, body mass index (BMI) and age at onset (rs = 0.408, -0.301 and -0.523 respectively, P＜0.05). Moreover, GAD-Ab wasthe only risk factor for predicting βcell failure in LADA patients (B = 1.455, EXP (B) = 4.283, P = 0.023). CONCLUSION: The decreasing rate of islet β cell function in LADA, being highly heterogeneous, is three times that of T2DM patients. The titer of GAD-Ab is an important predictor for the progression of islet β cell function, and age at onset and BMI could also act as the predictors.

MR diffusion-weighted imaging of rabbit liver VX-2 tumor

AIM: To investigate the implanting method of rabbit liver VX-2 tumor and its MR diffusion-weighted imaging (DWI) characteristics. METHODS: Thirty-five New Zealand rabbits were included in the study. VX-2 tumor was implanted subcutaneously in 14 rabbits and intrahepatically in 6 for pre-experiments. VX-2 tumor was implanted intrahepatically in 12 rabbits for experiment and three were used as the control group. DWI, T1- and T2-weighted of MRI were performed periodically in 15 rabbits for experiment before and after implantation. The distinction of VX-2 tumors on DWI was assessed by their apparent diffusion coefficient (ADC) values. The statistical significance was calculated by analysis of variance (ANOVA) of the randomized block design using SPSS10.0 software. RESULTS: The successful rate of subcutaneous implantation of VX-2 tumor was 29% (4/14) while that of intrahepatic implantation of it was 33% (2/6) in the preexperiment. The successful rate of intrahepatic implantation of VX-2 tumor in the experiment was 83% (10/12) and 15 tumors grew in 10 successfully implanted rabbits. The DWI signal of VX-2 tumor was high and became lower when the b value increased step by step. The signal of VX-2 tumor on the map of ADC was low. When the b value was 100 or 300 s/mm2, the ADC value of normal group and VX-2 tumor group was respectively 2.57±0.26, 1.73±0.31, 1.87±0.25 and 1.57±0.23 mm2/s. Their distinction was significant (F= 43.26, P<0.01), the tumor ADC value between b values 100 and 300 s/mm2 was significant (Tukey HSP,P<0.05) and the ADC value between VX-2 tumor and normal liver was also significant (Tukey HSP, P<0.01). VX-2 tumor developed quickly and metastasized early to all body, especially to the lung, liver, lymph nodes of mediastinum, etc. CONCLUSION: The DWI signal of rabbit VX-2 tumor has its characteristics on MR DWI and DWI plays an important role in diagnosing and discovering VX-2 tumor.

Induction of Heat Shock Protein 72 in RGCs of Rat Acute Glaucoma Model after Heat Stress or Zinc Administration

Purpose:To investigate the dynamics of heat shock protein 72(HSP72) expression in retinal ganglion cells(RGCs) in rat model of acute glaucoma treated with heat stress or intraperitoneal injection of zinc sulfate.Methods:Twenty-seven male Wistar rats were used to make acute glaucoma models. Five others served as normal control. Acute glaucoma models were made by intracameral irrigation in the right eyes with balanced salt saline (BSS) at 102 mmHg for 2 hours. Nine model rats were killed at different intervals after intracameral irrigation without treatment,which served as damage control. Ten were treated with heat stress 40℃～42℃, and 8 were used for zinc sulfate administration 2 days posterior to intracameral irrigation. Treated model rats were sacrificed at designed intervals after treatment. Right eyes were enucleated immediately, and the retinas were dissected for Western blot.Results:No HSP72 was found in RGCs of normal Wistar rats. In damage control group, slight HSP72 was detected during 6～36 hours posterior to intracameral irrigation. HSP72 was detected significantly expressed in RGCs of both heat shock group and zinc sulfate group. But the dynamics of HSP72 production were quite different in these two treated groups. In heat shock group, HSP72 appeared at the sixth hour after treatment, and increased gradually until its peak production emerged at the 48th hour. HSP72 vanished 8 days later after treatment. In zinc sulfate group, HSP72 expression began 24 hours later after zinc administration, and reached its highest level at the 72th hour posterior to treatment. HSP72 expression then decreased slowly, and disappeared 21 days later after treatment. Conclusion:HSP72 can be induced in RGCs of rat acute glaucoma models with heat stress or zinc sulfate adddministration. But the dynamics of the HSP72 induction in those two groups were quite different. Eye Science 2004;20:30-33.

Matrix Metalloproteinases Expression in Choroidal Neovascular Membranes

Purpose: To investigate the expression of matrix metalloproteinases (MMPs) in choroidal neovascular membranes with age-related macular degeneration (AMD).Methods: Seventeen choroidal neovascular membranes surgically removed from AMD patients with pars plana vitrectomy and subretinal membranes peeling were investigated. The expression of MMP-2 and MMP-9 was determined with immunohistochemical technique. Results: Immunohistochemistry staining in choroidal neovascular membranes for MMP-2 and MMP-9 was observed in 17 specimens. There was no detective of MMP-2 and MMP-9 in normal retinas. Conclusions: MMP-2 and MMP-9 were found in choroidal neovascular membranes, may degrade the Bruch membrane and be associated with the perforation of new vessels into Bruch membrane, involving a basic pathogenic process of AMD.

Study on the Difference in Expression of VEGF and bFGF and their Receptors in Young and Postmenopausal Women with Breast Cancer

OBJECTIVE To study the difference in expression of VEGF and bFGF and their receptors in young and postmenopausal women with breast cancer,METHODS Immunohistochemical methods (SABC) were used to study the expression of VEGF, FLK-1, bFGF and FLG on paraffin-embeddedsections from 40 cases of young and 30 cases of postmenopausal women with breast cancer, The relationship between axillary lymph node metastasis and the expression of the growth factors and their receptors was studied.RESULTS The mean expression of VEGF and bFGF and the positive rate of axillary lymph nodes in the young group were higher than that in the postmenopausal group (P<0.01 or P<0.05 ); the mean values of VEGF,bFGF, FLK-1 and FLG in cases of axillary lymph node metastasis were higher in patients without axillary lymph node metastasis in each group (P<0.05 or P <0.01); there was a significant difference between the meanexpression of VEGF. bFGF, FLK-1 and FLG in cases of stage 0-Ⅱ comparaed to cases of stage Ⅲ-Ⅳ (P<0.05 or P<O.01).CONCLUSION The tumor vasculature is directly related to the high breast cancer aggressiveness in young women, a characteristic that might be due to the high expression of VEGF and bFGF.

The preliminary efficacy of interferon-α and ribavirin combination treatment of chronic hepatitis C in HIV-infected patients

Background It is internationally accepted that in drug-naieve individualswith hepatitis C virus (HCV) and human immunodeficiency virus (HIV) co-infection, chronic hepatitisC should be treated first if the CD4 cell count does not require the initiation of anti-retroviraltherapy. Present paper evaluated the clinical effect and side-effect of interferon-α (IFN-α) andribavirin ( RBV) combination therapy for Chinese patients with HCV-HIV co-infection, and comparedwith them for HIV infection alone. Methods Ten patients with HCV-HIV and 17 patients with HCVreceived 5 million unit IFNα-2b every other day intramuscularly, and 300 mg RBV triple daily byoral. Dynamic observations were made for HCV RNA and HIV RNA loads, CD4^+ and CDS^+ T lymphocytecounts, liver function and blood cell measurement, and the medicine side-effects. Results After12-week and 24-week treatments of IFN-α and RBV combination therapy, mean HCV RNA levels reduced 1.14 logs and 1. 56 logs from the baseline at week 0 in HCV-HIV co-infection, and reduced 1. 48 logsand 1. 75 logs in HCV infection, respectively. The HIV RNA levels decreased 1. 22 logs and 1. 32logs from the base line; however, there were no obvious different changes at T lymphocyte counts ofHCV-HIV and HCV patients through 24-week treatments. Whole 27 patients showed satisfactorybiochemical response to therapy. There were some mild or mediate influence-like symptoms, intestinaluncomfortable and depressed blood cell counts in early stage of the treatments. No neuropsychiatricand auto-immune disorders were found. Conclusions IFN-α and RBV combination therapy had similaranti-HCV effects during 24-week treatment for HCV-HIV and HCV infected Chinese patients, and someanti-HIV effect. There were no obvious different biochemical responses and side-effects between twogroups above.