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Objective:To analyze the diagnostic value of sex hormone combined with carcinoembryonic antigen on lymph node metastasis in breast cancer patients.Methods:52 cases of breast cancer patients who underwent surgical resection were collected and divided into non-metastasis group and metastases group after axillary lymph node ultrasonography. The blood samples was collected from patients and the levels of serum estradiol, testosterone, progesterone and carcinoembryonic antigen were detected; The diagnostic value of the above indexes in patients with lymph node metastasis was analyzed; Logistic risk regression model was used to analyze the independent risk factors for lymph node metastasis after surgical resection.Results:There were significant differences between the non-metastatic group and the metastatic group in the largest tumor diameter and menopause (all P<0.05), but there were no significant differences among other general data (all P>0.05). The serum estradiol level in the non-metastatic group was (153.97±35.55) pg/ml, the progesterone level was (0.33±0.05) ng/ml, and the carcinoembryonic antigen level was (11.44±3.77) ng/ml, while the estradiol level in the metastatic group was (207.19±52.11) pg/ml ( t=4.13, P<0.001), progesterone level (0.38±0.04) ng/ml ( t=4.01, P<0.001), carcinoembryonic antigen level (15.41±3.46) ng/ml ( t=3.94, P<0.001). The above three indicators were significantly increased in patients in the transfer group. The area under the curve of estradiol was 0.83, the area under the curve of progesterone was 0.80, the area under the curve of carcinoembryonic antigen was 0.77, the area under the curve of the combination of the three was 0.85, and the area under the curve of the combination of the three was the largest. Logistic risk regression model showed that estradiol, progesterone, and carcinoembryonic antigen levels were independent risk factors affecting lymph node metastasis in breast cancer patients (all P<0.05) . Conclusion:The sex hormone estradiol, progesterone combined with carcinoembryonic antigen has a high diagnostic value for lymph node metastasis in patients with breast cancer, and can independently predict the occurrence of lymph node metastasis in breast cancer patients.
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Objective:The effect of intermedin (IMD) on ATP-induced activation of inflammatory bodies and pyroptosis of cells and its mechanism were studied using lipopolysaccharide (LPS)-sensitized mouse macrophage line RAW 264.7.Methods:The cells were divided into the control groups, the LPS groups, LPS+IMD groups, and LPS+IMD+LY294002 groups. The expression of interleukin (IL)-1β and IL-18 and the activation of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammatory cells were detected by real-time PCR and western blotting, and the pyroptosis of cells was detected by propidium iodide (PI) staining. The measurement data was represented by MS± SD, and the inter-group difference was compared with ANOVA calculations, and P<0.05 represented the difference with statistical significance. Results:Compared with the control group [(0.83±0.09) vs (0.49±0.04)], the ratio of phosphorylated phosphatidylinositol-3-kinase, p-PI3K)/phosphatidylinositol-3-kinase (PI3K) (0.44±0.05) and p-Akt/Akt (0.27±0.06) in the LPS group was significantly decreased. The ratios of p-PI3K/PI3K (1.22±0.18) and pAkt/Akt (0.83±0.09) in LPS+IMD group was significantly increased ( F=31.40, P<0.001; F=50.88, P<0.001). Compared with the control group, the mRNA and protein expressions of IL-1β, IL-18 and NLRP3 inflammasome (NLRP3, caspase-1, ASC) in RAW264.7 cells were up-regulated in the LPS group (LPS and ATP). Compared with LPS group, IMD treatment inhibited the expression of inflammatory cytokines IL-1β, IL-18 and NLRP3 inflammasome, which was blocked by LY294002, a blocker of PI3K/Akt pathway. The results of real-time PCR showed that the relative expression of IL-1β mRNA was (1.00±0.11) in the control group, (8.32±0.61) in the LPS group, (8.32±0.55) in the LPS+IMD group, and (7.23±0.41) in the LPS+IMD+LY group ( F=15.42, P<0.001). The relative expression of IL-18 mRNA in the control group was (1.00±0.17), (1.82±0.21) in the LPS group, (1.14±0.15) in the LPS+IMD group, and (1.53±0.11) in the LPS+IMD+LY group respectively ( F=18.16, P<0.001). The relative expression of NLRP3 mRNA in the control group was (1.00±0.13), (2.58±0.18) in the LPS group, (1.07±0.17) in the LPS+IMD group, and (1.33±0.32) in the LPS+IMD+LY group respectively ( F=15.98, P< 0.001); The relative expression of caspase-1 mRNA in the control group was (1.00±0.09), (6.20±0.19) in the LPS group, (3.43±0.06) in the LPS+IMD group, and (5.50±0.45) in the LPS+IMD+LY group respectively ( F=18.39, P<0.001). The relative expression of ASC mRNA in the control group was (1.00±0.21), (4.58±0.48) in the LPS group, (2.07±0.51) in the LPS+IMD group, and (3.33±0.32) in the LPS+IMD+LY group respectively ( F=15.19, P<0.001). Western blotting results showed that the relative expression of IL-1β protein was as follows: (100%) in the control group, [(188±14)%] in the LPS group, [(112±11)%] in the LPS+IMD group, and [(171±27)%] in the LPS+IMD+LY group respectively ( F=21.25, P<0.001). The relative expression of IL-18 protein in the control group was 100%, [(183±16)%] in the LPS group, [(115±19)%] in the LPS+IMD group, and [(179±23)%] in the LPS+IMD+LY group respectively ( F=19.62, P<0.001). The relative expression of NLRP3 protein was 100% in the control group, [(149±15)%] in the LPS group, [(106±10)%] in the LPS+IMD group, and [(144±15)%] in LPS+IMD+LY group respectively ( F=14.35, P<0.001). The relative expression of ASC protein was 100% in the control group, [(188±12)%] in the LPS group, [(110±18)%] in the LPS+IMD group, and [(192±8)%] in the LPS+IMD+LY group ( F=15.79, P<0.001). Conclusion:IMD inhibits the activation of NLRP3 inflammasome and cell pyroptosis by regulating PI3K/Akt activity.
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Objective:To investigate the expression of DNA damage repair factor DNA damage-binding protein 1 (DDB1) in hepatocellular carcinoma tissues, and the effect of DDB1 gene silencing on DNA repair and targeted killing in human hepatocellular carcinoma SMMC-7721 cells.Methods:The UALCAN platform was used to perform bioinformatics analysis on the expression of DDB1 in hepatocellular carcinoma tissues (371 cases) and paracancerous tissues (50 cases) in The Cancer Genome Atlas (TCGA) database and the correlation of DDB1 expression with the overall survival of liver cancer patients were analyzed by bioinformatics using the UALCAN platform. SMMC-7721 cells were transfected with small interfering RNA (siRNA) targeting DDB1 and negative control siRNA, which were DDB1 silencing group and negative control group, respectively. X-ray irradiation induced exogenous DNA double strand break (DSB) damage in the two groups of cells. Immunofluorescence staining (γH2AX was used for assessing cellular DSB damage, RPA32s33 and Rad51 were used for assessing homologous recombination repair) and Western blotting (were used to detect the level of RPA32s33 protein) were used to analyze the effect of DDB1 gene silencing on DSB damage repair. Sister chromosome exchange (SCE) experiment was used to analyze the frequency of SCE of homologous recombination of cells in DDB1 silencing group and negative control group. Tetramethylazozolium salt (MTT) method was used to analyze the killing effect of PARP inhibitor olapani (10 μmol/L), cisplatin (1 μg/ml) and olapani combined with cisplatin on SMMC-7721 cells in DDB1 silencing group and negative control group.Results:Bioinformatics analysis showed that the level of DDB1 mRNA in liver cancer tissues was higher than that in paracancerous tissues ( P < 0.001), and the overall survival of patients with high expression of DDB1 was worse than that of patients with low expression of DDB1 ( P = 0.029). When cultured for 4 hours after X-ray irradiation, the number of γH2AX foci in cells of the negative control group had mostly disappeared, and there were still more cells in DDB1 silencing group [(5.1±2.0) per cell vs. (13.4±2.0) per cell, t = -5.08, P = 0.007]. When cultured for 4 hours after X-ray irradiation, the number of RPA32s33 foci in the negative control group [(30.8±5.0) per cell vs. (13.2±1.6) per cell] and the number of Rad51 foci [(19.5±1.8) per cell vs. (8.3±3.3) per cell] were higher than those in the DDB1 silencing group, and the differences between the two groups were statistically significant (both P < 0.01). The frequency of SCE in the negative control group was higher than that in the DDB1 silencing group [(21.2±3.0)% vs. (11.2±1.6)%, t = 5.07, P = 0.007]. MTT assay showed that after olaparib treatment, the survival rate of cells in the DDB1 silencing group was lower than that in the negative control group [(40.3±3.6)% vs. (79.8±1.3)%, t = 17.94, P < 0.001]. When treated with olapani combined with cisplatin, the survival rate of cells in the two groups further decreased, and the survival rate of cells in the DDB1 silencing group was lower than that in the negative control group [(10.2±2.8)% vs. (29.6±3.4)%, t = 7.72, P = 0.002]. When treated with cisplatin alone, there was no significant difference in cell survival between DDB1 silencing group and negative control group [(41.9±5.1)% vs. (49.8±3.3)%, t = 2.71, P = 0.054]. Conclusions:The high expression of DDB1 in hepatocellular carcinoma tissues may be an important factor in the treatment resistance and poor prognosis of hepatocellular carcinoma. Knockdown of DDB1 gene expression can promote the sensitivity of SMMC-7721 cells to PARP inhibitors, and its mechanism may be related to the homologous recombination repair defect of SMMC-7721 cells caused by DDB1 silencing.
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ObjectiveTo investigate the anti-tumorigenesis function of rhDCN on the leukemia K562 cells in vitro and analyze the possible mechanism.Methods Exponential phase of K562 cells were transfected with pcDNA3.1(+)-DCN,and PBS,liposome alone,and pcDNA3.1(+) vector were as control groups.Morphology change of K562 cells was detected by Wright stain,and cell proliferation activity was detected by MTT. Cell cycle and apoptosis of K562 cells were assessed by FCM. The expression of apoptosis-related protein,including bcl-xl,Mcl-1 and Bax were detected by Western blot.ResultsWright stain showed that typical apoptotic morphology of K562 cells were observed in DCN transfected group.There were no morphological changes of apoptosis in other groups. MTT method results showed that proliferation inhibition rate of the transfected cells [24 h (16.14±1.08) ％,48 h (14.07±1.01) ％,72 h (20.29±1.19) ％]was higher than that of the other control groups (P ＜ 0.05).FCM results showed that the apoptosis index (20.15±1.31) ％of the DCN transfected group was higher than that of the other groups (P ＜ 0.05),and most of cells arrested in the G0/G1 phase (51.15±0.57) ％ (P ＜ 0.05).Western blot results showed the expression levels of Bax were increased while bcl-xl and Mcl-1 were decreased in pcDNA3.1(+)-DCN/K562 group.ConclusionrhDCN can inhibit the growth of K562 cells and induce the apoptosis.The effect of DCN on bcl-xl,Mcl-1,Bax may play a role in its mechanism.
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ObjectiveTo investigate the value for the clinical application of urokinase combined with Sodium Tanshion ⅡA in the treatment of cerebral thrombosis.Methods160 cases with cerebral thrombosis in our hospital were chosen.They were randomly divided into A group with 80 cases and B group with 80 cases.A group was the observation group (Tanshion ⅡA Urokinase combined treatment group),B group was the control group (urine kinase treatment group).2 weeks treatment was a course.The efficacy of treatment in the two groups at the end was taken for statistical analysis.ResultsThe neurological deficits were significantly improved in A and B group before and after treatment (P ＜ 0.05,there was statistically significant).The neurological deficit scores in A group after treatment was significantly lower than group B ( P ＜ 0.05,there was statistically significant).The total effective rate was 95 ％ by the treatment of urokinase and Tanshion Ⅱa in A group which was higher than group B ( P ＜ 0.05,there was statistically significant difference).ConclusionThe treatment of urokinase combined with Sodium Tanshion Ⅱa could improve neurological deficits and improve the clinical therapeutic effect.
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Objective To investigate the relationship between infection of helicobaeter pylori(Hp) and iron deficiency anemia(IDA) and to explore effective clinical treatments for patients with Hp associated IDA . Method 1. The Hp infection ratio of 40 chronic gastritis with IDA and 42 patients without IDA were counted up respectively. 2. 36 patients with Hp-positive chronic gastritis were divided into two groups randomly. One group was treated by using Hp eradication therapy in conjunction with oral iron supplement and the other using iron supplement only. The hema-tological parameters before and after treatment are measured and the effectiveness of the treatment was evaluated. Re-sult The Hp infection ratio in chronic gastritis patients with IDA is |figher than that in patients without IDA and the difference is significant. After Hp eradication therapy in conjunction with iron supplement Hb. serum iron and serum ferritin level in Hp associated IDA patients are increased significantly while that for patients treated by using iron sup-plement only have no significant improvement. Conclusion It appears that Hp infection may be related to IDA. When iron supplement treatment has no obvious effect to an IDA patient,lt may suggest a ease of Hp associated IDA. The i-ton supplement treatment has positive effects to IDA patients after Hp eradication.
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Objective To investigate the influence of different diluents(physiologic saline, dis-tilled water and human inactivated serum) on measurement of 15 items of biochemical parameters.Methods Fifteen items of biochemical parameters [alanine transaminase(ALT), aspartic transaminase (AST), alkaline phosphatase( ALP), gamma glutamyl transpeptidase ( GGT), creatine kinase ( CK),lactate dehydrogenase(LD), hydroxybutyrate dehydrogenase(HBDH), total bilirubin(TBIL), direct bilirubin(DBIL), total bile acid(TBA), ereatinine(Cr), uric acid(UA), eholesterol(CHO), glucose (GLU), and blood urea nitrogen(BUN)] were chosen. For each parameter, 45 serum samples with different eoncentrations of the parameter were collected. After diluted with different diluents(physio-logic saline, distilled water or human inactivated serum), the serum samples were detected by applying the fully automated biochemical analyzer. The mean value was calculated and statistical analysis was performed. Results There were some differences of detection results when the specimens were diluted with different diluents. ALT, AST, GGT, DBIL, and HBDH serum samples could be diluted by 10 times with physiologic saline, distilled water or human inactivated serums ALP and TBA serum sam-ples could only be diluted with inactivated serum, otherwise its result would be lower; GLU, TBIL samples could be diluted with distilled water and inactivated serums for BUN, CR, UA, CK, LDH,and CHO samples, physiologic saline or human inactivated serum might be optimal; if distilled water was chosen, the results of other parameters tented to decline except UA. It was BUN was improper to dilute the BUN samples with distilled water. In addition, there was no significant difference between the items diluted by 5 times and 10 times with physiologic saline. All the 15 items could be diluted with inactivated serum. Conclusion The inactivated serum should be the first choice of diluents to e-nure the accurate results of biochemical parameters. If the prepared inactivated serum is absent, we may choose other diluents according to the above-mentioned results.
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The search for a nonthrombogenic material with the potential for use in small diameter vascular graft applications continues to be a field of extensive investigation. This article describes the choice of biomaterials used as vascular prosthesis, the innovation of construction of tissue-engineered blood vessels, the indispensability, methods and the effect produced by surface modification of vascular prosthesis. The article also points out that research achievements of vascular prosthesis must be made with the exploitation of new nonthrombogenic biomaterial and the development of tissue engineering.
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Biocompatible Materials , Bioprosthesis , Blood Vessel Prosthesis , Prosthesis Design , Surface Properties , Tissue EngineeringABSTRACT
Objective To clone human neuron-specific enolase (NSE)gene and prepare the monoclonal antibodies against human neuron-specific enolase and to test the expression of NSE in tumor cell lines by immunocytochemistry (ICC).Methods The gene fragment of human NSE was amplified by RT-PCR and ligated to the pGEM vector. After the sequencing of recombinant NSE, it was ligated to the expression vector pMS-31b. The MS2-NSE fusion protein was expressed after higher temperature induction. The purified target protein was used for immunizing BALB/C mice to prepare McAbs against NSE.Results Full length of NSE gene with 1 305 bp was cloned. Molecular weight of MS2-NSE was 57 000. 1.42 mg/L of MS2-NSE fusion protein could be expressed. Two strains of hybridoma secreting NSE McAbs were obtained by ELISA screening. The subtypes of the NSE McAbs were IgG1and IgG2a. The two McAbs could react with A549 cell lines in ICC. NSE positive staining in ICC was mainly located in cytoplasm.Conclusions We clone human neuron-specific enolase gene, obtain the anti-NSE monoclonal antibodies and examine the expression of NSE in lung cancer tumor cell line.