Background and purpose:As a member of the catenin family, Delta-catenin protein could promote proliferation and invasion of tumor cells, but the accurate mechanism of Delta-catenin promoting cell proliferation is not clear. In the present study, we illustrated that Delta-catenin’s effect on cell apoptosis and their relationship with mitogen-activated protein kinase (MAPK) signaling pathway, and the possible mechanism was also explored for Delta-catenin promoting invasion and proliferation of tumor cells. Methods:The alterations of p38 and c-jun N-terminal rinasel JNK protein activity were detected in SPC and SK lung cancer cell lines with Delta-catenin overexpression or not, by Western blot method. At the same time, the apoptotic number of tumor cells was also examined by FCM method. Furthermore, the number of invasive tumor cells was examined by Matrigel invasive experiment. Results:Compared with untreated group and empty vector group, the activity of p38 protein was unchanged in lung cancer cell lines with Delta-catenin overexpressed (P>0.05), but the activity of JNK protein was decreased signiifcantly (P<0.05),meanwhile, apoptotic proportion of tumor cells were also reduced (P<0.05), and invasive ability of tumor cells was enhanced signiifcantly (P<0.05). Conclusion:Delta-catenin probably decreases apoptosis number of lung cancer cells via inhibiting the activity of JNK pathway, and then promotes invasive ability of tumor cells.

ObjectiveTo investigate the role of Notch1 gene in xenografted human cutaneous squamous cell (SCL-1) carcinoma. MethodsFifteen nude mice were divided into three groups, including untreated group(inoculated with SCL-1 cells treated with phosphate buffered saline), empty vector group (inoculated with SCL-1 cells transfected with empty vector) and Notch1 group(inoculated with SCL-1 cells transfected with Notch1 expression vector). All the mice were inoculated with SCL-1 cells(1 x 108/ml) of0.2 ml. Then, the growth of xenografted tumor was observed every other day. Fifteen days later, the mice were sacrificed, tumor tissue was dissected and subjected to terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay for the detection of cell apoptosis, reverse-transcription(RT)-PCR and Western blot for the examination of mRNA and protein expressions of Notch1, bcl-2 and bax, respectively. ResultsThe proliferation of xenografted tumor in Notch1 group was obviously inhibited compared with the untreated group. The weight of xenografted tumor in Notch1 group was significantly lower than that in the untreated group and empty vector group (0.574 ± 0.219 g vs. 2.642 ± 0.404 g and 2.606 ± 0.512 g, F= 26.642, P＜ 0.01). TUNEL assay demonstrated that the number of apoptotic cells per 500 cells in tumor tissue specimens was(87 ± 9) in Notch1 group, evidently higher than that in the untreated group(8 ± 2) and empty vector group(10 ± 3) (F = 194.266, P ＜ 0.05 ). Further, RT-PCR and Western blot revealed that the mRNA and protein expressions of Notch1 and bax were significantly upregulated, but those of bcl-2 were markedly downregulated in the Notch 1 group, with significant difference among the three groups(all P ＜ 0.05). ConclusionsNotch 1 gene can inhibit the growth of xenogra ffted human cutaneous squamous cell(SCL-1) carcinoma and induce SCL-1 cell apoptosis likely by upregulating bax expression and downregulating bcl-2 expression.

A glucose biosensor was fabricated from a glucose oxidasE-immobilized by chitosan and oxygen electrode. The effects of concentration of chitosan(0.3%), enzyme loading(0.8 mg), pH 7.0, phosphate buffer concentration(300 mmol/L), and temperature 25 ℃ for the response of the biosensor were investigated. The glucose biosensor has a linear response range of 0.016-1.10 mmol/L with a detection limit of 8.0 μmol/L(S/N=3). The response time was less than 60 s. The biosensor showed extremely good stability with a shelf-life of at least 3 months. The biosensor exhibited good repeatable response to a 0.25 mmol/L glucose olution with a relative standard deviation of 2.5%(n=10). The precision of fabrication of the biosensors using four different membranes was good with a RSD of 4.7%. Some common potential components in sample such as niacinamide, vitamin B6, vitamin B12, vitamin E, Ca2+, Mg2+, K+ and Zn2+ showed no interferences on the response of the glucose biosensor. The biosensor was successfully applied to determine the glucose in commercial beverage samples.

Objective To discuss the clinical efficacy of closed reduction and plaster splint fixation ( CRPSF) in the treatment of gerontal patients with distal radius fractures ( DRF) .Methods 76 elderly patients with DRF who treated by CRPSF were selected .According to AO classification of fractures ,the patients were divided into three groups,the A group had 27 cases;B group had 26 cases,C group had 23 cases.The treatment effect was evalua-ted by analyzing the follow-up data,the corresponding imaging measurement parameters and clinical scores .Results All patients had 12-month clinical and imaging follow-up.In the last follow-up, the arm-shoulder-hand dysfunction score and palmar angle ,ulnar deviation angle of A and B group were significantly better than those of C group , the differences were statistically significant(all P<0.05),the difference between A and B group was not statistically sig-nificant (P>0.05).In the last follow-up,the satisfaction score of C group was slightly lower than that of the A or B group,but had no statistically significant difference (P>0.05).Conclusion CRPSF in the treatment of gerontal pa-tients with DRF has good function and imaging effects ,and the improvement level has a certain relationship with the degree of fracture,but has no significant impact on the patients'satisfaction.

Objective To investigate the effect of pituitary tumor-transforming gene (PTTG)siRNA on the growth,invasion of,and expression of metastasis-related cytokines including matrix metalloproteinase-2 (MMP-2)and MMP-9 in xenografted human cutaneous squamous cell carcinoma in nude mice.Methods SCL-1 cells were subcutaneouslv inoculated into Balb/c nude mice to establish a xenograft model of human cutaneous squamous cell carcinoma.Then,15 mice bearing xenografted carcinoma were equally divided into 3 groups to be inoculated with phosphate buffer saline (PBS),control siRNA,and PTTG siRNA of 50 nmoI/L,respectively,ever),other day for 2 weeks.The size of xenograted carcinoma in these mice was measured every other day.At the end of 2-week treatment.the mice were killed followed by the evaluation of tumor weight,as well as the quantification of mRNA and protein expression of PTTG,MMP-2 and MMP-9 by reverse transcription (RT)-PCR and Western-blot,respectively.Results The xenograft model of human cutaneous squamous cell carcinoma was successfully established.The treatment with PTTG siRNA obviously inhibited the growth of the xenografted tumom and the expression of PTTG mRNA and protein compared with PBS and control siRNA (all P<0.05).In addition,the expression of MMP-2 and MMP-9 in xenografted tumors in PTTG siRNAtreated mice were significantly lower than those in PBS and control siRNA-treated mice.suggesting that PTTG siRNA evoked the decrease in invasive and metastatic ability of xenografted tumors.Conclusions PTTG siRNA can inhibit the growth of human cutaneous squamous cell carcinoma xenografts in nude mice,and downregulate the expression of invasion-and metastasis-related cytokines,including MMP-2 and MMP-9.

ObjectiveTo investigate the role and clinical pathological significance of PTTG and bFGF in cutaneous squamous cell carcinoma(CSCC).MethodsTissue specimens were collected from the lesions of 42 patients with CSCC and normal skin of 42 normal human controls.The protein and mRNA expressions of PTTG and bFGF were detected by immunohistochemistry and in situ hybridization in these specimens respectively.ResultsA significant increase was observed in the positive expression rates of PTTG and bFGF proteins[64.3％(27/42) vs.11.9％(5/42),73.8％(31/42) vs.21.4％(9/42),both P＜ 0.05] and mRNA [59.5％(25/42) vs.7.1％(3/42),75.0％(29/42) vs.16.7％(7/42),both P＜ 0.05] in the CSCC tissue specimens than in the control specimens.The protein and mRNA expressions of PTTG were positively correlated with those of bFGF(both P ＜ 0.05),and closely correlated with histological grade of CSCC (both P ＜0.05).ConclusionThe high expression of PTTG and bFGF may be associated with the initiation of CSCC.

[ ABSTRACT] AIM:To investigate the effect of DEK downregulation on the apoptosis of gastric carcinoma SGC-7901 cells, and to explore its associations with NF-κB signaling pathway and apoptosis related proteins.METHODS:SGC-7901 cells with different treatments were divided into 3 groups including untreated group, control siRNA group and DEK siRNA group.The expression of DEK at mRNA and protein levels in the SGC-7901 cells was detected by real-time PCR and Western blot.The cell apoptosis was examined by flow cytometry.Furthermore, the activities of caspase-3 and caspase-9 in the SGC-7901 cells were investigated by Caspase-Glo?-3/9 kit.Finally, the expression of key regulatory pro-tein p65 of NF-κB signaling pathway and apoptosis-related proteins Bcl-2 and Bax in the SGC-7901 cells was investigated by Western blot.RESULTS:Compared with untreated group and control siRNA group, the expression of DEK at mRNA and protein levels was significantly downregulated in DEK siRNA group (P<0.05).In addition, the ratios of early phase apoptosis and total apoptosis in DEK siRNA group were markedly higher than those in untreated group and control siRNA group (P<0.05).Most notably, the decrease in p65 and Bcl-2 proteins, increase in Bax protein and the increases of caspase-3 and caspase-9 activities were observed in DEK siRNA group.CONCLUSION:Downregulation of DEK mediates cell apoptosis of gastric carcinoma may be tightly associated with NF-κB signaling pathway.

Traditional Chinese medicine has been paid more and more attention in the development of modern healthcare,and its clinical diagnosis and treatment have broad prospects and enormous potential.However,the current traditional Chinese medicine diagnosis and treatment model have serious shortcomings in service capacity and,diagnosis,and treatment effect.The rapid development of big data and artificial intelli-gence technology provides an opportunity for the iterative upgrade of traditional Chinese medicine diagnosis and treatment models.This article reviewed the current situation of artificial intelligence empowering tradi-tional Chinese medicine clinical diagnosis and treatment,clarified the problems and challenges faced by artifi-cial intelligence technology in data integration,data quality,and data analysis in traditional Chinese medicine clinical diagnosis and treatment,and proposed to empower from the aspects of disciplinary integration,data quality optimization,data privacy protection,and promotion and application,so as to provide reference for im-proving the effectiveness of traditional Chinese medicine clinical diagnosis and treatment.

Objective To evaluate the effect of downregulation of microRNA (miR)-373 expression on cell cycle and apoptosis of a cutaneous squamous cell carcinoma (CSCC) cell line A431.Methods A431 cells at exponential growth phase were classified into 3 groups:miR-373 inhibitor group and negative control group transfected with miR-373 inhibitor and negative control miRNA respectively,and untreated group receiving no treatment.At 48 hours after the transfection,real-time PCR was performed to determine the expression of miR-373 in the above 3 groups,cell counting kit-8 (CCK-8) assay to evaluate the effect of downregulated expression of miR-373 on the proliferation of A431 cells,flow cytometry to investigate the distribution of cell cycle and changes in apoptosis of A431 cells in different treatment groups,and colorimetric analysis to detect the changes in caspase-3 activity in different treatment groups.Statistical analysis was carried out with SPSS 17.0 software by using two-sample t test for the comparison between two groups,one-way analysis of variance (ANOVA) for the comparison among 3 groups,and least significant difference (LSD)-t test for multiple comparisons.Results The expression of miR-373 was significantly lower in the miR-373 inhibitor group (0.120 ± 0.036) than in the untreated group (1.002 ± 0.022) and negative control group (1.037 ± 0.028,LSD-t =36.21,34.83,respectively,both P ＜ 0.001).At 48,72 and 96 hours,the miR-373 inhibitor group showed significantly decreased proliferative activity of A375 cells compared with the untreated group and negative control group (F =10.805,13.720 and 30.907 respectively,P =0.038,0.010 and 0.001 respectively).The proportion of A375 cells in G0/G1 phase was significantly higher in the miR-373 inhibitor group (64.69％ ± 1.18％) than in the untreated group (52.74％ ± 0.66％,t =15.51,P ＜ 0.001) and negative control group (53.80％ ± 0.80％,t =13.24,P ＜ 0.001).The proportion of total apoptotic cells and activity of caspase-3 in the miR-373 inhibitor group were 22.69％ ± 1.24％ and 1.238 ± 0.057 respectively,which were significantly higher than those in the untreated group (9.62％ ± 1.14％,0.413 ± 0.028 respectively,both P ＜ 0.001)and negative control group (9.66％ ± 0.97％,0.437 ± 0.036 respectively,both P ＜ 0.001).Conclusion MiR-373 may play an important role in the regulation of cell cycle and induction of apoptosis of the CSCC cell line A431.

The purpose was to optimize the vitrification for porcine embryos cryopreservation. Blastocyst/Morula (5-6th day-embryos) were collected from superovulated Bama mini-pigs (sows/gilts). We compared different cryopreservation methods, cryopreservation tools, thining of zona pellucida (ZP) and recipient breeds on the efficiency of porcine embryo cryopreservation. The results showed that: in embryo survival rate and blastocyst cell number, there were no significant differences between cryopreservation method I [embryos were vitrified by two step method with open pulled straw (OPS) and glass micropipette (GMP) in solution 1 (TCM199 + 20% FBS + 10% EG + 10% DMSO) for 3 min, and solution 2 (TCM199 + 20% FBS + 20% EG + 20% DMSO + 0.4 mol/L SUC) for 1 min, stored in liquid nitrogen] and method II[Blastocysts were cultured for 25 min in NCSU23 + 7.5 microg/mL cytochalasin B, centrifuged at approximately 13 000 xg for 12-13 min, and recovered back into pNCSU23. They were then equilibrated for 5 min in 2 mol/L ethylene glycol in pNCSU23, washed quickly in the vitrification medium, 8 mol/L ethylene glycol, 7% polyvinylpyrrolidone (PVP) in pNCSU23, loaded into OPS/GMP, and plunged into liquid nitrogen]. GMP vitrification method was more suitable and efficient than OPS method (P < 0.05) in embryo survival rate (83.8% vs 77.6%) and blastocyst cell number (53.1 vs 47.5) after thawing. Thining of ZP did not increase the survival rate, but significantly improved blastocyst cell number in the survival blastcysts (60.1 and 46, P < 0.01). Local pig breeds (Fengjing sows) were more suitable as recipients for embryo transfer of vitrified/warmed blastcysts, which can improve pregnant rate and embryo efficiency.
Animals ; Blastomeres ; cytology ; Cryopreservation ; methods ; veterinary ; Embryo Transfer ; veterinary ; Embryo, Mammalian ; Swine ; Swine, Miniature ; Vitrification ; Zona Pellucida ; physiology