Objective To investigate the pro-apoptotic effects of an ?1-adrenergic inhibitor,doxazosin,in HeLa cells and the role of transcription factor activator protein-2? (AP-2?) in the process.Methods The HeLa cells were treated with doxazosin (0,10,40 or 60 ?mol/L) for 16 h,and cell apoptosis was detected by flow cytometric analysis.After doxazosin (60 ?mol/L) treated the HaLa cells,the cells transfected with AP-2? overexpressing constructs or an antisense oligonucleotide against AP-2? for 16 h,the apoptosis was analyzed with FCM.The expression of AP-2? and caspase-3 in above 3 groups of cells was detected by relative quantitative RT-PCR and Western blot analysis,respectively.Results Doxazosin increased the apoptotic rate and total cell death rate of the HeLa cells,and upregulated the expressions of AP-2? and caspase-3 in a dose-dependent manner.Overexpressing AP-2? improved the increased apoptotic rate and total cell death rate induced by doxazosin,and enhanced the increased expression of caspase-3.Whereas doxazosin-induced apoptosis and the total cell death in HeLa cells were decreased by antisense AP-2?,and antisense AP-2? in part abolished the increased effects of doxazosin on caspase-3 expression.Conclusion Doxazosin induces apoptosis in HeLa cells in a dose-dependent manner,and transcription factor AP-2? is functionally involved in this process.

Objective To study the function of let-7 and miR-24 in UVB-induced apoptosis.Methods After NIH3T3 cells were irradiated with UVB.Hoechest33342/PI staining was used to study the cell apoptosis and RT-PCR was used to uralyzc the expression level of let-7 and miR-24.In addition,potential target genes of these miRNAs in PicTar were classified into different function categories through GOstat software.Results Compared to the control,the NIH3T3 cells exposure to UVB appeared typical apoptotic and necrotic ceils by fluorescence microscope.The exprossion level of let-7 and miR-24 in NIH3T3 cells after UVB irradiation was higher than that of the control.Among the target genes,casp3,bc1212,map3kl and cdk5 were also involved in UVB-induced apoptosis mechanism.Conclusion Let-7 and miR-24 play a role in UVB-induced apoptosis.

Objective To explore the effects of ultraviolet on DNA damage in keratinocytes and to observe the protective role of resveratrol for the cells. Methods Comet assay was employed to evaluate the damage after radiation with different doses of UV rays (UVA, UVB and UVC) of 0, 10, 30, 50, 70 and 90 mJ/cm2, and the effects after pretreatment with various concentrations of resveratrol under irradiation with 30 mJ/cm2. Results UVA irradiation (0 ~ 90 mJ/cm2) had no significant effects on HaCaT cells. However, TailDNA%, TailLength, CometLength, TailMoment and OliveTailMoment showed both UVB and UVC induced DNA damage in a dose-de-pentent manner. UVC was more harmful than UVB at the same dose. Conclusions The DNA breakage induced by UVB and UVC is dose-dependent. As compared with UVB, UVC is more harmful to HaCaT cells. Resveratrol exerts a protective effect in HaCaT cells irradiated by UVB or UVC.

Objective To investigate the effects of hyperbaric oxygen preconditioning (HBOP) on brain edema, inflammatory reaction and neuronal cell apoptosis induced by experimental hemorrhage in rats. Method Eighteen male Spraque-Dawley rats, weighing 300 - 350 g,received five successive sessions of HBOP with 3 atmosphere absolute pressure and 100% O2 one hour daily for five successive days, and other eighteen rats received five successive sessions of pretreatment with one atmosphere absolute pressure, air, one hour daily for five successive days. Twenty-four hours after the final pre-conditioning, rats received an infusion of 100 μL autologous blood into the basal ganglion. Seventy-two hours later, rats were sacrificed for brain edema measurements in 12 rats of each group. The histopathological changes around the hematoma were observed microscopically, and the neuronal cell apoptosis was detected by using the terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) in six rats of each group. Data of brain water content were analyzed by using Stata 7.0 software and statistical analysis was carried out by two-tailed Student t -test. Results Compared with the control group, HBOP significantly attenuated brain edema 72 hours after intra-cerebral hemorrhage in experimental rats (81. 6± 0. 7% vs. 82. 8± 0.9%, P < 0.01). Inflammatory cell infiltration and neuronal cell apoptosis were also significantly decreased in the HBOP group. Conclusions HBOP protects the rats against brain edema formation, and quells inflammatory reaction and neuronal cell apoptosis following intra-cerebral hemorrhage in experimental rats.

Objective: To observe the expression of Smad in diabetic rat’s kidney and the effect of BaoYuan II to it.Methods: SD rats were randomly divided into four groups: control group,model group,Benazepril-treatment group and BaoYuan II-treatment group.The latter three groups were induced to be diabetic nephropathy by intraperitoneal injection with streptozotocin and then the drugs were treated respectively.After eight weeks,the plasma albumin,blood glucose,urinary protein,urinary?2-M were determined and expression of mRNA of TGF-?1,Smad2,Smad3,Smad7 was detected by RT-PCR.Results: Compared with diabetic rats group,the level of plasma albumin was increased and the level of blood glucose,urinary protein,urinary?2-M was depressed in BaoYuan II-treatment group.The expression of TGF-?1 and Smad2,Smad3 were down-regulated and Smad7 was up-regulated in model group.The effects of BaoYuan II were similar to Benazepril.Conclusion: BaoYuan II had protected effect to diabetic nephropathy rats.It acted by regulating TGF-?1 and Smad2,Smad3,Smad7 possibly.

AIM: To investigate the effect of 3-methyladenine (3-MA) combined with trichostatin A (TSA) on triple-negative breast cancer cells and the mechanism involved.METHODS: The viability of MDA-MB-231 cells was detected by MTT assay, the migration ability was determined by scratch assay, and the expression of autophagy-related proteins was detected by Western blot.RESULTS: TSA significantly inhibited the viability of MDA-MB-231 cells in a time-and dose-dependent manner.The results of scratch assay showed that TSA inhibited cell migration ability.Western blot data indicated that TSA resulted in a moderate increase in LC3-Ⅱ expression.Moreover, 3-MA inhibited cell autophagy induced by TSA, and combination of 3-MA and TSA further inhibited the viability of MDA-MB-231 cells.CONCLUSION: Combination of 3-MA and TSA may effectively inhibit the growth of triple-negative breast cancer cells.

Objective To investigate the carcinogeic role of miR-365 in cuntanerous squamous cell carcinoma (cSCC).Methods Normal HaCaT cells were divided into control and irradiation groups,the later was exposed by UVB irradiation (50 J/m2).MicroRNA expression profiles of the two groups were analyzed by microRNA array.The expression variations of miR-365 in HaCaT,A431,Tca8113 and HSC-1 cells were validated by qRT-PCR analysis.The colony-forming and invasion capacities were dectected by colony forming assay and Transwell migration assay in vitro,respectively.HaCaTpre-miR365-2 highly expressing miR-365 was constructed by retroviral vector infection.Tumorigenicity evaluation was carried out by subcutaneously inject of the cells at the right back flank of nude mice.Results There were 30 microRNAs differentially expressed in HaCaT cells after UVB irradiation and miR-365 was one of the most sensitive miRNAs(as high 6.7 times as control).Expression of miR-365 in all the cSCC cell lines A431,Tca8113 and HSC-1 were significantly higher than that in HaCaT cell,in which the maximum was A431 (15.67 ±1.12 times,P ＜ 0.01),and the minimum was TcaS113 (4.72 ± 0.85 times,P ＜ 0.05).Knockdown of miR-365 in cSCC cell lines significantly inhibited the colony forming ability (t =13.68,P ＜ 0.05) and cell migration (t =19.98,P ＜ 0.05) in vitro.HaCaT cells overexpressing miR-365 by transient transfection significantly increased the ability of colony formation (t =7.11,P ＜ 0.05) and cell migration (t =22.03,P ＜0.05) in vitro.In addition,HaCaTpre-miR-365-2 cell line stably expressing miR-365 could successfully establish tumors in nude mice.Conclusions MiR-365 is an oncogene for cutaneous squamous cell carcinoma.

Objective To study the expression level of miR-21 in UVB irradiated HaCaT cells and A431 cells. Methods Real-time qPCR was used to examine the expression level of miR-21 in HaCaT cells and A431 cells after 50 J/m2UVB radiation. The possible target genes were predicted by PicTar and performed function categories with Gostat analysis. Results Compared with the HaCaT cells, miR-21 the expression level in A431 cells increased over 4 times. At 2h and 4h after UVB irradiation, the expression level of miR-21 in HaCaT cells were up regulated, and it lowered 2 times at 8 h compared with the control.There was no further change in the expression level of miR-21 after 12 h. While miR-21 expression levels in A431 cells were not changed signifcantly. The results of target prediction and Gostat analysis suggested that PIK3R1, BCL2 and E2F3 were involved in the cell differentiation and cell process. Conclusion miR-21 possibly involved in the pathogenesis of epidermal squamous cell carcinoma and the mechanism of UVB-induced injury.

Objective To establish the model and software for quality assessment of fetal nuchal translucency ultrasound image using computer image recognition technology.Methods The proposed approach firstly divided the input image into four sub-image blocks:the nasal bone(NB) area,the nuchal translucency (NT) area,the midbrain area,and the jaw and chest area.For each sub-image block,the algorithm compared the image block with the corresponding area of the standard training image set,and then determined whether the current image block was the qualified one using the the Gabor feature and Bayesian decision.The input ultrasound image was determined to be qualified only if it had four qualified sub-image blocks.Results The difference between our automatic method and the manual screening by experts wasting small,the method obtained Kappa =0.795 and P ＜0.001.Moreover,the efficiency of our method was much higher than the manual screening method.Conclusions Image recognition technology can effectively assist the sonographer to assess the quality of fetal NT of ultrasound image.The proposed approach can reduce the subjectivity and randomness of the manual evaluation of NT image.

The infection of HBV may cause acute and chronic hepatitis B,and potentially lead to hepatocirrhosis and liver cancer.As a defense mechanism of organism to resist external infection,RNA interference(RNAi) has become a powerful tool for us to study its effects on antiviral infection and gene therapy in recent years.In this article we summarize the mechanisms of RNA interference and the progress on anti-HBV infection studies by RNAi.