Management of anastomotic biliary stricture through utilizing percutaneous transhepatic cholangioscopy.

AIM: Occurrence of anastomotic biliary stricture (AS) remains an essential issue following hepatobiliary surgeries, and percutaneous transhepatic cholangioscopy (PTCS) has great therapeutic significance in handling refractory AS for patients with altered gastrointestinal anatomy after cholangio-jejunostomy. This present study aimed to investigate feasibility of PTCS procedures in AS patients for therapeutic indications. MATERIALS AND METHODS: This study was a single-center, retrospective cohort study with a total number of 124 consecutive patients who received therapeutic PTCS due to AS. Clinical success rate, required number, and adverse events of therapeutic PTCS procedures as well as patients survival state were reviewed. RESULTS: These 124 patients previously underwent choledochojejunostomy or hepatico-jejunostomy, and there was post-surgical altered gastrointestinal anatomy. Overall, 366 therapeutic PTCS procedures were performed for these patients through applying rigid choledochoscope, and the median time of PTCS procedures was 3 (1-11). Among these patients, there were 34 cases (27.32%) accompanied by biliary strictures and 100 cases (80.65%) were also combined with biliary calculi. After therapeutic PTCS, most patients presented with relieved clinical manifestations and improved liver functions. The median time of follow-up was 26 months (2-86 months), and AS was successfully managed through PTCS procedures in 104 patients (83.87%). During the follow-up period, adverse events occurred in 81 cases (65.32%), most of which were tackled through supportive treatment. CONCLUSION: PTCS was a feasible, safe and effective therapeutic modality for refractory AS, which may be a promising alternative approach in clinical cases where the gastrointestinal anatomy was changed after cholangio-jejunostomy.

Exosomes derived from TGF-ß1-pretreated mesenchymal stem cells alleviate biliary ischemia-reperfusion injury through Jagged1/Notch1/SOX9 pathway.

BACKGROUND: This study aimed to evaluate the efficacy of exosomes (EXO) derived from TGF-ß1-pretreated mesenchymal stem cells (MSCs) on biliary ischemia reperfusion injury (IRI) and further reveal the possible mechanisms. METHODS: Bone marrow-derived MSCs were treated with exogenous TGF-ß1, Jagged1/Notch1/SOX9 pathway inhibitor LY450139, or their combination. Then, EXO were isolated from the culture supernatants and further characterized. After establishing IRI model of biliary epithelial cells (EpiCs), EXO derived from differently-treated MSCs were applied to detect their protective effects on EpiCs, and LY450139 was applied in EpiCs to detect the possible mechanisms after treatment with MSCs-EXO. EXO derived from differently-treated MSCs were further injected into the hepatic artery immediately after establishment of intrahepatic biliary IRI for animal studies. RESULTS: Pretreatment with TGF-ß1 significantly enhanced MSCs-EXO production and elevated the levels of massive miRNAs associated with anti-apoptosis and tissue repair, which were evidently decreased after TGF-ß1 plus LY450139 cotreatment. Notable improvement was observed in EpiCs after MSCs-EXO treatment, evidenced by reduced cellular apoptosis, increased cellular proliferation and declined oxidative stress, which were more evident in EpiCs that were treated with EXO derived from TGF-ß1-pretreated MSCs. However, application of EXO derived from TGF-ß1 plus LY450139-cotreated MSCs reversely enhanced cellular apoptosis, decreased cellular proliferation and anti-oxidants production. Interestingly, LY450139 application in EpiCs after treatment with MSCs-EXO also reversed the declined cellular apoptosis and enhanced oxidative stress induced by TGF-ß1 pretreatment. In animal studies, administration of EXO derived from TGF-ß1-pretreated MSCs more effectively attenuated biliary IRI through reducing oxidative stress, apoptosis, inflammation and enhancing the expression levels of TGF-ß1 and Jagged1/Notch1/SOX9 pathway-related markers, which were reversed after administration of EXO derived from TGF-ß1 plus LY450139-cotreated MSCs. CONCLUSION: Our results provided a vital insight that TGF-ß1 pretreatment endowed MSCs-EXO with stronger protective effects to improve biliary IRI via Jagged1/Notch1/SOX9 pathway.

Assessing the efficacy of a recombinant H9N2 avian influenza virus-inactivated vaccine.

The H9N2 avian influenza virus has been widely spread in poultry around the world. It is proved to the world that the avian influenza virus can directly infect human beings without any intermediate host adaptation in "1997 Hong Kong avian influenza case," which shows that the avian influenza virus not only causes significant losses to the poultry industry but also affects human health. In this study, we aimed to address the problem of low protection of avian H9N2 subtype influenza virus vaccine against H9N2 wild-type virus. We have rescued the H9.4.2.5 branched avian influenza virus isolated in South China by reverse genetics technology. We have recombined these virus (rHA/NA-GD37 and rHA/NA-GD38) which contain hemagglutinin and neuraminidase genes from the H9N2 avian influenza virus (MN064850 or MN064851) and 6 internal genes from the avian influenza virus (KY785906). We compared the biological properties of the virus for example virus proliferation, virus elution, thermostability, and pH stability. Then, we evaluated the immune effects between rHA/NA-GD37 and GD37, which show that the recombinant avian influenza virus-inactivated vaccine can stimulate chickens to produce higher antibody titers and produce little inflammatory response after the challenge. It is noticeable that the recombinant virus-inactivated vaccine had better immune impact than the wild-type inactivated vaccine. Generally speaking, this study provides a new virus strain for the development of a H9N2 vaccine.

The epigallocatechin gallate derivative Y6 inhibits human hepatocellular carcinoma by inhibiting angiogenesis in MAPK/ERK1/2 and PI3K/AKT/ HIF-1α/VEGF dependent pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Hypervascularity has been considered as one of the major features of many solid tumors. Green tea is one of the commonly drink resources in China, and its active component, Epigallocatechin gallate (EGCG), exhibits antiangiogenic activities in various experimental tumor models. However, EGCG has many shortages, e.g., relatively unstable, low lipid solubility, poor bioavailability, and short duration of action. AIM OF THE STUDY: To overcome the shortages of EGCG for antiangiogenic antitumor usage, our study developed a novel EGCG derivate, Y6(5,3',4',3″,4″,5″-6-0-ethyl-EGCG). The underlying mechanism was also elucidated. MATERIAL AND METHODS: we evaluated the effects of EGCG, Y6 on HCC and angiogenesis in vivo and in vitro. Moreover, to understand their antitumor mechanisms, key factors within angiogenesis-related signaling pathways (MAPK/ERK1/2, PI3K/AKT, HIF-1 VEGF) were analyzed by using western blot, immunohistochemistry (IHC), quantitative real-time quantitative PCR (RT-PCR). HepG2 xenograft model and the chorioallantoic membrane (CAM) were used to investigate the effects of Y6 and EGCG on tumors and anti-angiogenesis in vivo. Micro-vessel density (MVD) was analyzed by IHC of CD34 staining. IHC, qRT-PCR and Western blot were used to detect the expression of HIF-1α and VEGF protein in tumor tissues. The protein levels of MAPK/ERK1/2, PI3K/AKT, HIF-1α, and VEGF in tumor tissues were detected by western blot. RESULTS: Our results demonstrated that both EGCG and Y6 displayed antiangiogenetic and antitumor effects against HCC cells in vitro and in vivo. We found that rather than equal amount of EGCG, Y6 displayed better abilities in inhibiting the growth of HCC tumor cells, as well as inhibiting the growth of neovascularization in the chick embryos and HepG2 xenograft tumors bearing-mice, based on the data obtained from MTT assay, immunohistochemistry (IHC), chick chorioallantoic membrane (CAM) assays. In the comparison of equivalent dose of EGCG, qRT-PCR data showed that Y6 induced more significant decrease of the mRNA levels of HIF-1α and VEGF in supernatant-treated SMMC-7721 cells under hypoxic condition, as well as in the in xenograft tumor tissues; whereas Y6 also significantly reduced the protein levels of MAPK/ERK1/2, PI3K/AKT, HIF-1α, and VEGF to a greater extent than EGCG, determined by western blotting assay. CONCLUSIONS: our work suggests that the new EGCG derivate Y6 could significantly inhibit tumor growth and angiogenesis which is possibly involved with the signaling intervention of MAPK/ERK1/2 and PI3K/AKT/HIF-1α/VEGF pathways, and is supposed to be a potential therapeutic reagent for anti-angiogenesis treatment of solid tumors.

[Testosterone supplementary therapy for type-2 diabetes mellitus males with hypogonadism: Controversy and analysis].

As more and more studies suggest that type 2 diabetes mellitus (T2DM) is closely related to male hypogonadism, people begin to pay more attention to the role of testosterone in the development of T2DM and the effect and safety of testosterone supplementary therapy. There is some controversy in randomized controlled studies and meta-analyses about the effects of testosterone supplementation on the blood glucose level, androgen deficiency symptoms, and cardiovascular diseases. This review focuses on the diagnosis of hypogonadism in T2DM males, differences in the therapeutic effects and safety of testosterone replacement among different studies, and rational use of testosterone supplementation for T2DM patients.

Serum Insulin-like growth factor-1 levels of healthy adults in southern China.

It is to establish the normal range and investigate the distribution characteristics of serum Insulin-like growth factor-1 (IGF-1) for healthy adults in southern China. IGF-1 levels of 515 healthy adults (254 males and 261 females) were measured by automated chemiluminescence immunoassay. The subjects were strictly selected healthy volunteers, aged 20 to 84 years old, with equal five year intervals and without abnormal conditions that impacted IGF-1 levels. The reference ranges were calculated using the smooth centile curves of the LMS method (L: coefficient of skewness, M: median, S: coefficient of variation). IGF-1 declined with aging in adults. There were statistically significant differences for the IGF-1 levels between men and women in some subgroups of age. Gender differences varied depending on the age. Middle-aged females had higher IGF-1 whilst elder females had lower IGF-1. The statistical differences were seen in three subgroups of age between this study and a German cohort that is the reference range for the laboratory test kit. Here, the age- and gender-specific normal range was established for Chinese adults. A Z Score of IGF-1 for an individual could be obtained via the LMSchartmaker application, which standardized IGF-1 research worldwide.

[Hypogonadism and the quality of life in male patients with type-2 diabetes mellitus].

OBJECTIVE: To compare the level of testosterone between type-2 diabetes mellitus (T2DM) patients and healthy controls and to investigate the status of hypogonadism and the influence of hypopgonadism on the quality of life. METHODS: We collected serum total testosterone (TT), free testosterone (FT), sex hormone-binding globulin (SHBG), and other clinical data from 166 T2DM patients aged over 30 years and 186 age-matched healthy controls. We investigated the quality of life (QoL) of the two groups of subjects using the questionnaires of Androgen Deficiency in Aging Males (ADAM), Aging Male Symptoms (AMS), 36-Item Short-Form Health Survey (SF-36), and Special Quality of Life for Diabetes Mellitus (DSQL). RESULTS: The level of calculated FT (cFT) was remarkably lower in the T2DM patients than in the healthy controls (P<0.05), but no statistically significant differences were observed between the two groups in the levels of TT, bio-available testosterone (Bio-T), and SHBG. The T2DM males with hypogonadism showed significant differences from those without in age, height, systolic blood pressure, and creatinine (P<0.05). Based on the criteria of cFT <0.3 nmol/L and AMS score ≥27, the incidence rate of hypogonadism was 51.81% in the T2DM patients, 31.58% in the 30－39 yr group, 32.50% in the 40－49 yr group, 50% in the 50－59 yr group, 69.23% in the 60－69 yr group, and 77.27% in the ≥70 yr group, elevated by 77.4% with the increase of 10 years of age (OR = 1.774, P<0.001). The AMS score was significantly correlated with the scores of DSQL (r = 0.557, P<0.001) and SF-36 (r = －0.739, P<0.001) in the T2DM patients. CONCLUSIONS: T2DM patients have lower levels of cFT than healthy men, accompanied with a higher incidence of hypogonadism. Age is a main risk factor of hypogonadism. Severer testosterone deficiency symptoms are associated with lower scores of QoL in T2DM males.

Enhancement of power production with tartaric acid doped polyaniline nanowire network modified anode in microbial fuel cells.

The feasibility to use tartaric acid doped PANI for MFC anode modification was determined. Uniform PANI nanowires doped with tartaric acid were synthesized and formed mesoporous networks on the carbon cloth surface. By using this tartaric acid doped PANI modified carbon cloth (PANI-TA) as the anode, the voltage output (435 ± 15 mV) and power output (490 ± 12 mW/m(2)) of MFC were enhanced by 1.6 times and 4.1 times compared to that of MFC with plain carbon cloth anode, respectively. Strikingly, the performance of PANI-TA MFC was superior to that of the MFCs with inorganic acids doped PNAI modified anode. These results substantiated that tartaric acid is a promising PANI dopant for MFC anode modification, and provided new opportunity for MFC performance improvement.

Microbial fuel cell-based biosensors for environmental monitoring: a review.

The microbial fuel cell (MFC) is an innovative technology that was initially designed to harness energy from organic waste using microorganisms. It is striking how many promising applications beyond energy production have been explored in recent decades. In particular, MFC-based biosensors are considered to be the next generation biosensing technology for environmental monitoring. This review describes recent advances in this emerging technology of MFC-based biosensors, with a special emphasis on monitoring of biochemical oxygen demand and toxicity in the environment. The progress confirms that MFC-based biosensors could be used as self-powered portable biosensing devices with great potential in long-term and remote environmental monitoring.

A whole-cell electrochemical biosensing system based on bacterial inward electron flow for fumarate quantification.

Fumarate is of great importance as it is an oncometabolite as well as food spoilage indicator. However, cost-effective and fast quantification method for fumarate is lacking although it is urgently required. This work developed an electrochemical whole-cell biosensing system for fumarate quantification. A sensitive inwards electric output (electron flow from electrode into bacteria) responded to fumarate in Shewanella oneidensis MR-1 was characterized, and an electrochemical fumarate biosensing system was developed without genetic engineering. The biosensing system delivered symmetric current peak immediately upon fumarate addition, where the peak area increased in proportion to the increasing fumarate concentration with a wide range of 2 µM-10 mM (R(2)=0.9997). The limit of detection (LOD) and the limit of quantification (LOQ) are 0.83 µM and 1.2 µM, respectively. This biosensing system displayed remarkable specificity to fumarate against other possible interferences. It was also successfully applied to samples of apple juice and kidney tissue. This study added new dimension to electrochemical biosensor design, and provide a simple, cost-effective, fast and robust tool for fumarate quantification.

Adsorption and removal of triphenylmethane dyes from water by magnetic reduced graphene oxide.

Triphenylmethane (TPM) dye is one of the most prevalent and recalcitrant water contaminants. Magnetic reduced graphene oxide (rGO) is an efficient adsorbent for organic pollutants removal. However, the performance and adsorption kinetics of magnetic rGO towards TPM have not yet been studied. In this study, a magnetic Fe3O4@rGO nano-composite, which could be easily removed from water with a simple magnetic separation step was synthesized and characterized. The magnetic rGO showed fast adsorption rate and high adsorption capacity towards different TPM dyes (the Langmuir monolayer adsorption capacity is 64.93 mg/g for adsorption of crystal violet). The adsorption processes are well-fitted to the pseudo-second-order kinetic model (R(2) > 0.99) and the Langmuir isotherm model (R(2) = 0.9996). Moreover, the magnetic rGO also showed excellent recycling and regeneration capabilities. The results indicated that adsorption with magnetic rGO would be a promising strategy to clean up the TPM contamination.

Enhanced bioelectricity generation by improving pyocyanin production and membrane permeability through sophorolipid addition in Pseudomonas aeruginosa-inoculated microbial fuel cells.

Improvement on electron shuttle-mediated extracellular electron transfer (EET) is of great potential to enhance the power output of MFCs. In this study, sophorolipid was added to enhance the performance of Pseudomonas aeruginosa-inoculated MFC by improving the electron shuttle-mediated EET. Upon sophorolipid addition, the current density and power density increased ∼ 1.7 times and ∼ 2.6 times, respectively. In accordance, significant enhancement on pyocyanin production (the electron shuttle) and membrane permeability were observed. Furthermore, the conditions for sophorolipid addition were optimized to achieve maximum pyocyanin production (14.47 ± 0.23 µg/mL), and 4 times higher power output was obtained compared to the control. The results substantiated that enhanced membrane permeability and pyocyanin production by sophorolipid, which promoted the electron shuttle-mediated EET, underlies the improvement of the energy output in the P. aeruginosa-inoculated MFC. It suggested that addition of biosurfactant could be a promising way to enhance the energy generation in MFCs.

8-60hIPP5(m)-induced G2/M cell cycle arrest involves activation of ATM/p53/p21(cip1/waf1) pathways and delayed cyclin B1 nuclear translocation.

Protein phosphatase 1 (PP1) is a major serine/threonine phosphatase that controls gene expression and cell cycle progression. The active mutant IPP5 (8-60hIPP5(m)), the latest member of the inhibitory molecules for PP1, has been shown to inhibit the growth of human cervix carcinoma cells (HeLa). In order to elucidate the underlying mechanisms, the present study assessed overexpression of 8-60hIPP5(m) in HeLa cells. Flow cytometric and biochemical analyses showed that overexpression of 8-60hIPP5(m) induced G2/M-phase arrest, which was accompanied by the upregulation of cyclin B1 and phosphorylation of G2/M-phase proteins ATM, p53, p21(cip1/waf1) and Cdc2, suggesting that 8-60hIPP5(m) induces G2/M arrest through activation of the ATM/p53/p21(cip1/waf1)/Cdc2/ cyclin B1 pathways. We further showed that overexpression of 8-60hIPP5(m) led to delayed nuclear translocation of cyclin B1. 8-60hIPP5(m) also could translocate to the nucleus in G2/M phase and interact with pp1α and Cdc2 as demonstrated by co-precipitation assay. Taken together, our data demonstrate a novel role for 8-60hIPP5(m) in regulation of cell cycle in HeLa cells, possibly contributing to the development of new therapeutic strategies for cervix carcinoma.

Inadequate glycaemic control and antidiabetic therapy among inpatients with type 2 diabetes in Guangdong Province of China.

BACKGROUND: Diabetes mellitus has become epidemic in recent years in China. We investigated the prevalence of hyperglycaemia and inadequate glycaemic control among type 2 diabetic inpatients from ten university teaching hospitals in Guangdong Province, China. METHODS: Inadequate glycaemic control in diabetic patients was defined as HbA1c = 6.5%. Therapeutic regimens included no-intervention, lifestyle only, oral antiglycemic agents (OA), insulin plus OA (insulin + OA), or insulin only. Antidiabetic managements included monotherapy, double therapy, triple or quadruple therapy. RESULTS: Among 493 diabetic inpatients with known history, 75% had HbA1c = 6.5%. Inadequate glucose control rates were more frequently seen in patients on insulin + OA regimen (97%) than on OA regimen (71%) (P < 0.001), and more frequent in patients on combination therapy (81% - 96%) than monotherapy (75%) (P < 0.05). Patients on insulin differed significantly from patients on OA by mean HbA1c, glycemic control rate, diabetes duration, microvascular complications, and BMI (P < 0.01). CONCLUSIONS: This study showed that glycaemic control of type 2 diabetic patients deteriorated for patients who received insulin and initiation time of insulin was usually delayed. It is up to clinicians to move from the traditional stepwise therapy to a more active and early combination antidiabetic therapy to provide better glucose control.

Multicenter clinical study on the efficacy and safety of inhalable insulin aerosol in the treatment of type 2 diabetes.

BACKGROUND: A new inhalable insulin aerosol (Inh-Ins) was developed in China. The aim of this multicenter clinical study was to evaluate the efficacy and safety of this new Inh-Ins as a treatment of type 2 diabetes. Regular porcine insulin (RI) was used as a control. METHODS: This study is a prospective, randomized, open-label, parallel-group multicenter clinical trial in which 253 qualified patients with type 2 diabetes received the insulin Glargine daily at bedtime plus either a pre-meal Inh-Ins or a pre-meal subcutaneous RI for 12 weeks. HbA1c, fasting plasma glucose (FPG), the 1-hour-postprandial blood glucose (1hPBG) and the 2-hour-postprandial blood glucose (2hPBG) were measured. Events were monitored for adverse effects. RESULTS: After 12 weeks, the HbA1c decreased significantly from baseline in both treatment groups, with no significant difference between the two regimens. In the Inh-Ins group, FPG, both 1hPBG and 2hPBG significantly declined from baseline after the 8th- and 12th-weeks of treatment. The reduced values of FPG or 1hPBG between the two groups showed a more significant hypoglycemic effect with the Inh-Ins than the RI. After 12 weeks, the pulmonary carbon monoxide diffusing capacity (DLco) was significantly lower in Inh-Ins group than in the RI. The main side effects of Inh-Ins were coughing, excessive sputum, and hypoglycemia. CONCLUSIONS: Inh-Ins was effective in decreasing HbA1c like the RI. It was better in lowering the FPG and the 1hPBG than the RI. Its main side effects were coughing, excessive sputum, and hypoglycemia. Also, Inh-Ins slightly impaired DLco.

[The comparison of insulin aspart and human soluble insulin used in insulin pump therapy in newly diagnosed type 2 diabetic patients].

OBJECTIVE: To compare the efficacy of insulin aspart and human soluble insulin used in insulin pump therapy on the islets beta cell function in newly diagnosed type 2 diabetic patients. METHODS: Fifty-nine hospitalized newly diagnosed type 2 diabetic patients, 35 males and 24 females, aged 51 +/- 12, were and randomly divided into 2 groups to undergo insulin pump therapy with insulin aspart (aspart group, n = 30) or human soluble insulin (human insulin group, n = 29) for 2 weeks. The targets of glycemic control included fasting blood glucose (FBG) < 6.1 mmol/L and 2 h postprandial blood glucose (PBG) < 8.0 mmol/L. The changes of blood glucose, and the time and the doses of insulin needed for good glycemic control were compared between the two groups. The frequency of hypoglycemia and pump-related side effects were recorded. RESULTS: On the 2nd day of insulin pump therapy, FBG and 3 meals PBG levels were significantly reduced in both groups while the post-breakfast and post-dinner blood glucose levels were far more decreased in the aspart group than in the human insulin group (8.4 mmol/L +/- 2.8 mmol/L vs 11.3 mmol/L +/- 3.8 mmol/L, and 9.0 mmol/L +/- 2.4 mmol/L vs 10.7 mmol/L +/- 2.8 mmol/L, both P < 0.05). The FBG and 3 meals PBG were significantly lowered in the aspart group than in the human insulin group on the 7th day and after the stopping of insulin pump therapy. The time of good glycemic control of the aspart group was 2.0 d, significantly shorter than that of the human insulin group (6.0 d, P < 0.01). The mean dose of insulin used during insulin pump therapy in the aspart group was 0.6 U/kg, significantly less than that in the human insulin group (0.8 U/kg, P = 0.002). There was no significant difference in the AIR, mean area under the curve (AUC) of insulin and C peptide during IVGTT, HOMA-beta and proinsulin between the two groups before and after insulin pump therapy. No pump-related side effects were observed in both groups. CONCLUSION: In newly diagnosed type 2 diabetic patients with short term insulin pump therapy, the use of insulin aspart was more effective and faster with less doses of insulin in acquiring good glucose control compared with humulin R.

[Apoptosis of hepatocarcinoma cell line HLE induced by the combination of wild type P53 gene and 1,2:5,6-dianhydro-3,4-diacetylgalactitol].

BACKGROUND & OBJECTIVE: P53 gene is a tumor suppressor gene,and its mutation in human tumor cells is frequently observed. Previous studies revealed that wild type p53 (wt-p53)gene can suppress proliferation ,and induce apoptosis of tumor cells. However,the enhancive effect of wt-p53 on apoptosis of tumor cells is not so obvious when it is used alone. Therefore,it is a new field for tumor research that wt-p53 gene combined with drug is used to enhance apoptosis rate of tumor cells. This study was to investigate the enhancement effect of the combination of wt-p53 and 1,2:5,6-dianhydro-3,4-diacetylgalactitol (DADAG)on apoptosis of human hepatocarcinoma cell line HLE. METHODS: HLE cells were transfected with pUHD10-3-p53 plasmid contained wt-p53 gene,and treated with DADAG. After 96-hour treatment,apoptosis was evaluated by flow cytometry and DNA electrophoresis. RESULTS: The apoptosis rates were: 1.4% in untreated group, 3.5% in pUHD10-3-transfection group, 32.6% in DADAG group, 43.4% in pUHD10-3-p53-transfection group, and 74.6% in pUHD10-3-p53-transfection plus DADAG-treatment group. DNA ladder was observed in pUHD10-3-p53-transfection plus DADAG-treatment group. CONCLUSION: Apoptosis of HLE cells could be induced by both wt-p53 gene and DADAG,and the effect was more obvious when HLE cells were treated by the combination of wt-p53 gene and DADAG.