Objective To explore the effect of HIPK2 on apoptosis of human kidney tubular epithelial cells (HKC) induced by cisplatin.Methods Apoptosis of HKC cells was induced by cisplatin and the expression of HIPK2 was detected by RT-qPCR and Western blot.Two HIPK2 siRNAs were designed according to gene sequence of HIPK2 and cell lines with HIPK2 knockdown were established through transfecting the HIPK2 siRNAs into HKC cells by liposome.The expression of HIPK2 mRNA and protein was detected by RT-qPCR and Western blot after induced by cisplatin.Then cell apoptosis was detected by Annexin V/PI after the HIPK2-knockdown cells were treated with cisplatin.Moreover,the expression of pro-apoptotic protein bax was detected by Western blot after HIPK2 was knockdown.Results The expression of HIPK2 mRNA and protein was down-regulated obviously on the process of HKC apoptosis which induced by dose-dependent cisplatin (P＜0.05).The transfection of siRNA could significantly reduce the expression of HIPK2 mRNA and protein in HKC (P＜0.05),which promotes the HKC cells apoptosis induced by cisplatin.Conclusions HIPK2 can suppress the HKC cells apoptosis induced by cisplatin.

Objective To investigate the effect of high dose mifepristone and high dose progesterone in the treatment of patients with endometrial carcinoma and to explore the possible mechanisms associating with them Methods Thirty untreated patients diagnosed as endometrial carcinoma through dilation and curettage of the uteri were divided into 3 groups at random Each group was given medroxyprogesterone acetate(MPA),(500 mg/day) or mifepristone(MIF),(100 mg/day)or MIF(100 mg/day)+ MPA(500 mg/day)for 5 days respectively On the sixth day, hysterectomy was performed on these patients The endometrial cancer specimen of post hysterectomy was compared with the one of pre administrating The morphologic changes of the endometrial cancer cells were observed through light microscope Immunohistochemistry assay (SP method) was applied to determine the localization and immunoreactive intensity of proliferating cell nuclear antigen(PCNA), estrogen receptor (ER), progesterone receptor (PR), B cell leukemia lymphoma 2 (bcl 2), bcl 2 associated X protein(bax) and CD 44 v6 Results Better differentiation degree and active excretion were observed in all of the post hysterectomy endometrial specimen In the same time, apoptosis of carcinoma cells was observed The most significant changes were seen in the MIF+MPA group In the MPA group,the pre treatment and post treatment expression of PR(2 9?1 1,1 6?0 8),ER(2 8?0 9,1 4?0 9),PCNA(0 84?0 10,0 60?0 12),bcl 2(0 236?0 089,0 157?0 981) and CD 44 v6 (4 6?1 8,2 5?1 9) were all decreased(all P 0 05) In the MIF+MPA group, the expression of PR(3 2?1 0,0 8?0 8),ER(2 7?0 9,0 7 ?0 9 ),PCNA(0 81?0 09,0 25?0 09),bcl 2(0 225?0 091,0 066?0 009)and CD 44 v6(4 5?1 9,2 7?1 6) were all decreased(all P

Objective To study the clinical significance of the expression of D-amino acid oxidase(DAO) in hepatocellular carcinoma(HCC) tissues.Methods The gene expression profiles and related clinical data of 214 HCC cases were collected.Univariate and multivariate analyses were conducted to investigate the association between DAO expression levels,clinical traits,the prognosis.Results Univariate analysis indicated that there were a lower level of blood alpha fetoprotein(AFP,P=0.001),a smaller number of nodules(P=0.042),a better TNM stage(P=0.014),a lower metastasis risk(P=0.001) and a better prognosis(P=0.011)in the samples with high DAO expression.Multivariate analysis also indicated a lower AFP level(OR:0.162,95%CI:0.078-0.336) and metastasis risk(OR:0.140,95%CI:0.069-0.284) in the samples with high DAO expression,as well as a better prognosis(OR:0.833,95%CI:0.700-0.992).Conclusion DAO expression was associated with blood AFP levels,metastasis risk and prognosis in HCC,and its high expression was a protective factor for HCC.

Objective To analyze the clinical effect of biliary-enteric anastomosis and biliary stent to palliative treatment of malignant obstructive jaundice.Methods A total of 40 patients with inoperable malignant obstructive jaundice were enrolled in this study,and 20 patients were performed biliary stent placement (stent group),simultaneously 20 patients were performed biliary-enteric anastomosis (operation group).The fatality rate after operation,the level of total bilirubin before treatment and after treatment for 4,7,14 d,the rate of hyperpyrexia,nausea and vomiting,postoperative recurrence of jaundice were compared between two groups.Results There were no dead in two groups.The level of total bilirubin was decreased after treatment,and there was no significant in two groups before treatment and after treatment for 4,7,14 d (P ＞ 0.05).The rate of hyperpyrexia in operation group was significantly lower than that in stent group (0 vs.4/20) (P ＜ 0.05).Conclusion For palliative treatment of malignant obstructive jaundice,the biliary-enteric anastomosis should be performed first if there is no significant contraindication.

10.3969/j.issn.2095-4344.2013.26.002

Objective To construct expression vectors of calmodulin(CaM)mutants N2 and C2,and to express,purify,and identify the mutant proteins,in order to study the interactions between CaM and calcium channels. Methods The cDNA of N?lobe and C?lobe of CaM were used to prepare the cDNA of N2 and C2. Next,the recombinant cDNAs were cloned into a pGEX?6p?3 plasmid,and the recombinant plasmids were trans?ferred into E.coli BL21 cells. The transfected BL21 cells were stimulated with IPTG. The fusion proteins were extracted by ultrasonication and puri?fied by using GS?4B beads. Finally,protein activity was identified by the pull?down assay. Results Both the restriction digestion map and the DNA sequence identification results confirmed that the recombinant plasmids were successfully constructed. SDS?PAGE results showed high purity and concentration of N2 and C2 proteins. Their activities and binding abilities with the calcium channel fragment were confirmed by the pull?down assay.Conclusion In this study,expression vectors of N2 and C2 are successfully constructed,and physiologically active N2 and C2 CaM mutant proteins are obtained.

Objective : To explore the association between coffee intake and all cancer mortality in East Asian populations. Methods : We searched literatures which assessed the relationship between coffee intake and cancer mortality in Asian populations published by December 10th,2018 from China National Knowledge Infrastructure,Wanfang Database,VIP Database and PubMed. We conducted category and dose-response meta-analyses using Stata 15.0. Results : A total of 335 relevant articles were retrieved; five articles were finally included in the meta-analyses,of which four were carried out in Japanese population and one in Singaporean Chinese population. The total sample size of the five articles was 361 802,and the number of deaths from cancer was 17 664. The results showed that coffee intake reduced the risk of all cancer mortality in East Asian populations（RR=0.93,95%CI：0.87-0.99）. There was no statistical significant association between coffee intake and all cancer mortality in East Asian men（RR=0.94,95%CI：0.77-1.15）. Among East Asian women,coffee consumption reduced the risk of all cancer mortality by 12%（RR=0.88,95%CI：0.81-0.95）. All cancer mortality risk decreased with the increase of coffee intake,and reached the lowest point at one and a half cups of coffee per day（RR=0.92,95%CI：0.86-0.98）. Conclusion Coffee intake reduced the risk of all cancer mortality in East Asian populations,which was obviously found in East Asian women. Drinking one and a half cups of coffee a day had the lowest risk of all cancer mortality.

Objective: To explore the molecular mechanism of miR-200b-3p regulates the proliferation, invasion, migration and apoptosis of pancreatic cancer cells by targeting vascular endothelial growth factor A (VEGFA). Methods: The expression of miR-200b-3p in pancreatic cancer tissues and cell lines was detected by quantitative real-time fluorescence PCR (qRT-PCR). Pancreatic cancer PANC-1 cells were divided into NC group, miR-200b-3p mimic group, si-VEGFA group and si-VEGFA+ miR-200b-3p inhibitor group. The proliferation, migration and invasion of PANC-1 cells were measured by CCK-8 and Transwell assay. The apoptosis of PANC-1 cells was detected by Annexin V-FITC/PI double staining flow cytometry assay. The targeted relationship of miR-200b-3p and VEGFA was estimated by dual luciferase reporter gene assay and Western blotting. Results: The expression of miR-200b-3p in pancreatic cancer tissues and cell lines was decreased. After miR-200b-3p was overexpressed in PANC-1 cells for 48 h, the cell viabilities of PANC-1 cells in NC group and miR-200b-3p mimic group were 1.250±0.028 and 0.983±0.044, the numbers of migrated cells were 402.700±21.530 and 158.000±17.620, the numbers of invaded cells were 478.300±31.050 and 170.000±32.470, and the cell apoptosis rates were (5.280±0.352)% and (7.430±0.393)%. The cell viability, migration and invasion of PANC-1 cells in miR-200b-3p mimic group were significantly decreased than those in NC group (t=5.060, P=0.007; t=8.796, P=0.001; t=6.863, P=0.002). The cell apoptosis rate in miR-200b-3p mimic group was significantly higher than that in NC group (t=4.076, P=0.015). The fluorescence intensity in VEGFA-WT group was 1.000±0.027, which was significantly higher than that in VEGFA-WT+ miR-200b-3p mimic group (0.632±0.048; t=6.637, P=0.003). The fluorescence intensities in VEGFA-MUT group and VEGFA-MUT + miR-200b-3p mimic group were 1.000±0.049 and 0.868±0.047, with no statistically significant difference (t=1.944, P=0.124). After miR-200b-3p was overexpressed in PANC-1 cells for 48 h, the expressions of VEGFA in NC group and miR-200b-3p mimic group were 1.000±0.058 and 0.762±0.020, respectively. The expression level in miR-200b-3p mimic group was lower than that in NC group (t=3.908, P=0.017). After transfection of PANC-1 cells with si-VEGFA or si-VEGFA + miR-200b-3p inhibitor for 48 h, the cell viabilities of PANC-1 cells in NC group, si-VEGFA group and si-VEGFA + miR-200b-3p inhibitor group were 1.300±0.058, 0.943±0.047 and 1.143±0.023, the numbers of migrated cells were 446.000±17.350, 206.300±19.360 and 428.300±30.330, and the numbers of invaded cells were 510.300±24.550, 175.700±24.290 and 473.700±35.530, with statistically significant differences (F=15.830, P=0.004, F=33.530, P=0.001, F=38.860, P<0.001). The cell viability, migration and invasion of PANC-1 cells in si-VEGFA group were significantly decreased than those in NC group (P=0.003, P<0.001, P<0.001). There was no significant difference between si-VEGFA + miR-200b-3p inhibitor group and NC group (P=0.107, P=0.854, P=0.671). The cell apoptosis rates in NC group, si-VEGFA group and si-VEGFA+ miR-200b-3p inhibitor group were (3.810±0.577)%, (7.373±0.482)% and (3.650±0.514)%, with a statistically significant difference (F=16.020, P=0.004). The cell apoptosis rate in si-VEGFA group was significantly higher than that in NC group (P=0.007), but there was no significantly difference between si-VEGFA + miR-200b-3p inhibitor group and NC group (P=0.975). Conclusion miR-200b-3p suppresses the proliferation, invasion and migration and promotes the apoptosis of pancreatic cells by down-regulating VEGFA.

OBJECTIVETo compare quantitatively the enhanced thin CT section with pathologic findings in pulmonary carcinoma, pulmonary inflammatory pseudotumor (IPT) and pulmonary tuberculoma so as to demonstrate the relation of degree of enhancement and the vascular structure within the lesion with special emphasis on pulmonary carcinoma.

METHODSEnhanced thin CT sections were obtained in 35 cases with nodular or patchy lesions in the peripheral lung field which are difficult to differentiate clinically. There were pulmonary carcinoma 21, inflammatory pseudotumor 7 and tuberculoma 7. The number of small vessels (inner diameter 0.02 approximately 0.1 mm), relatively large vessels (inner diameter > 0.1 mm) and their vascular bed areas were analyzed by computed image analyzing system. The relation between CT average attenuation and the number of vessels or the vascular bed areas were statistically evaluated.

RESULTS1. The differences of average attenuation in carcinoma, inflammatory pseudotumor and tuberculoma were statistically significant (P < 0.05). 2. The differences in number of small vessels, relatively large vessels and vascular bed areas among these three types of lesion were also significant (P < 0.05). 3. A positive correlation was found in the average CT affenuation of lung carcinoma and its number of small vessels and relatively large vessels and 4. A positive correlation was found between the average CT attenuation in these three lesions and the relatively large vessels, total vascular amount and vascular bed areas.

CONCLUSIONS1. The average degree of attenuation, being divided into four degrees, is of practical value in the differentiation of lung carcinoma, inflammatory pseudotumor and tuberculoma. 2. The average CT attenuation of lung carcinoma, inflammatory pseudotumor and tuberculoma is in direct proportion to the number of vessels and vessel bed areas and 3. The characteristic CT enhancement in lung carcinoma reflexes the condition of vessels and blood supply within the tumor.

Adult ; Aged ; Female ; Humans ; Lung ; blood supply ; diagnostic imaging ; pathology ; Lung Neoplasms ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Plasma Cell Granuloma, Pulmonary ; diagnostic imaging ; pathology ; Tomography, X-Ray Computed ; methods ; Tuberculoma ; diagnostic imaging ; pathology ; Tuberculosis, Pulmonary ; diagnostic imaging ; pathology

Objective To investigate whether human RhoA is modified by SUMO. Methods Overlap extension PCR and double digestion technique were used to construct the eukaryotic expression vector pcDNA3-3flag-RhoA, which was identified by sequencing. The plasmid was transfected into HEK293T cells and its expression was detected by Western blotting. Immunofluorescence assay was used to detect whether RhoA is co-localized with SUMO. Co-Immunoprecipitation was used to detect whether RhoA is modified by SUMO. Results The recombinant plasmid pcDNA3-3flag-RhoA was successfully constructed and verified. Western blotting showed that the recombinant plasmid pcDNA3-3flag-RhoA expressed abundant fusion protein in HEK293T cells. Immunofluorescence showed that RhoA was co-localized with SUMO2/3 but not with SUMO1. Co-immunoprecipitation verified that RhoA was modified by SUMO2/3 but not SUMO1. Conclusion Human RhoA is modified by SUMO2/3 and probably participates in the regulation of axon regrowth after nervous system injury.