Atomically Co-dispersed nitrogen-doped carbon for sensitive electrochemical immunoassay of breast cancer biomarker CA15-3.

The development of an ultrasensitive and precise measurement of a breast cancer biomarker (cancer antigen 15-3; CA15-3) in complex human serum is essential for the early diagnosis of cancer in groups of healthy populations and the treatment of patients. However, currently available testing technologies suffer from insufficient sensitivity toward CA15-3, which severely limits early large-scale screening of breast cancer patients. We report a versatile electrochemical immunoassay method based on atomically cobalt-dispersed nitrogen-doped carbon (Co-NC)-modified disposable screen-printed carbon electrode (SPCE) with alkaline phosphatase (ALP) and its metabolite, ascorbic acid 2-phosphate (AAP), as the electrochemical labeling and redox signaling unit for sensitive detection of low-abundance CA15-3. During electrochemical detection by differential pulse voltammetry (DPV), it was found that the Co-NC-SPCE electrode did not have a current signal response to the AAP substrate; however, it had an extremely favorable response current to ascorbic acid (AA). Based on the above principle, the target CA15-3-triggered immunoassay enriched ALP-catalyzed AAP produces a large amount of AA, resulting in a significant change in the system current signal, thereby realizing the highly sensitive detection of CA15-3. Under the optimal AAP substrate concentration and ALP catalysis time, the Co-NC-SPCE-based electrochemical immunoassay demonstrated a good DPV current for CA15-3 in the assay interval of 1.0 mU/mL to 10,000 mU/mL, with a calculated limit of detection of 0.38 mU/mL. Since Co-NC-SPCE has an excellent DPV current response to AA and employs split-type scheme, the constructed electrochemical immunoassay has the merits of high preciseness and anti-interference, and its clinical diagnostic results are comparable to those of commercial kits.

Characteristics of recurrence, predictors for relapse and prognosis of rapid relapse triple-negative breast cancer.

Background: Triple-negative breast cancer (TNBC) patients who recur at different times are associated with distinct biological characteristics and prognoses. Research on rapid-relapse TNBC (RR-TNBC) is sparse. In this study, we aimed to describe the characteristics of recurrence, predictors for relapse, and prognosis in rrTNBC patients. Methods: Clinicopathological data of 1584 TNBC patients from 2014 to 2016 were retrospectively reviewed. The characteristics of recurrence were compared between patients with RR-TNBC and slow relapse TNBC(SR-TNBC). All TNBC patients were randomly divided into a training set and a validation set to find predictors for rapid relapse. The multivariate logistic regression model was used to analyze the data of the training set. C-index and brier score analysis for predicting rapid relapse in the validation set was used to evaluate the discrimination and accuracy of the multivariate logistic model. Prognostic measurements were analyzed in all TNBC patients. Results: Compared with SR-TNBC patients, RR-TNBC patients tended to have a higher T staging, N staging, TNM staging, and low expression of stromal tumor-infiltrating lymphocytes (sTILs). The recurring characteristics were prone to appear as distant metastasis at the first relapse. The first metastatic site was apt to visceral metastasis and less likely to have chest wall or regional lymph node metastasis. Six predictors (postmenopausal status, metaplastic breast cancer,≥pT3 staging,≥pN1 staging, sTIL intermediate/high expression, and Her2 [1+]) were used to construct the predictive model of rapid relapse in TNBC patients. The C-index and brier score in the validation set was 0.861 and 0.095, respectively. This suggested that the predictive model had high discrimination and accuracy. The prognostic data for all TNBC patients showed that RR-TNBC patients had the worst prognosis, followed by SR-TNBC patients. Conclusion: RR-TNBC patients were associated with unique biological characteristics and worse outcomes compared to non-RR-TNBC patients.

Risk factors of non-sentinel lymph node metastasis in 443 breast cancer patients with sentinel lymph node-positive.

Axillary lymph node dissection is the standard surgical procedure for breast cancer patients with sentinel lymph node (SLN) positive. In clinical practice, axillary lymph node dissection may be an unnecessary treatment for some breast cancer patients with non-sentinel lymph node (NSLN) negative. The aim of this study was to analyze the risk factors of NSLN metastasis in breast cancer patients with SLN positive. Four hundred fifty-six clinical early stage breast cancer patients with SLN positive were collected and analyzed in the oncological surgery department of Fujian Provincial Hospital during 2013 to 2018. All these patients underwent surgical treatment. The average age and tumor size of 443 patients with SLN positive breast cancer were (49.8 ± 10.8) years and (2.42 ± 0.94) cm. Univariate analysis showed that the size of primary tumor, the number of positive SLN, the number of negative SLN, the ratio of positive SLNs, and the type of metastases in SLN were the influencing factors of NSLN metastasis. Multivariate regression analysis showed that primary tumor size T > 2 cm (P < .001, OR = 2.609), the positive number of SLNs ≥3 (P = .002, OR = 5.435), the ratio of positive SLNs ≥ 50% (P = .017, OR = 1.770), and SLN macrometastases (P < 0.001, OR = 16.099) were independent risk factors for NSLN metastasis. Combined with the 4 independent risk factors, the area under the curve to predict NSLN metastasis was 0.747 > 0.7. For clinical early breast cancer with positive SLN, primary tumor size T > 2 cm,the positive number of SLNs ≥ 3, the ratio of positive SLNs ≥ 50%, and SLN macrometastases could predict NSLN metastasis well, and guide surgery to avoid overtreatment.

Risk Factors of Brain Metastasis and Prognosis in HER2-Positive Breast Cancer: A Single-Institution Retrospective Analysis from China.

Background: Brain metastasis (BM) frequently occurs in HER2-positive breast cancer (BC) patients, but the risk factors of BM in this type of patients are still unknown. Our study aims to assess the risk factors of BM and prognostic analysis in HER2-positive BC patients. Methods: Univariate analysis used t-test, chi-square test, and Fisher's exact test to find out the risk factors for BM, and multivariable analysis was done with stepwise logistic regression analysis. Prognostic data analysis was estimated by the Kaplan-Meier method. Results: A total of 228 HER2-positive BC patients were included, of whom 214 patients were postoperative metastatic patients and 14 patients were de novo stage IV patients. Through comparing the stratified variables between 51 postoperative metastatic patients with BM and 163 postoperative metastatic patients without BM, the multivariate analysis showed that age ≤40 years (OR 2.321, 95% CI: 1.089 to 4.948) and first metastatic site with lung metastasis (OR 2.168, 95% CI: 1.099 to 4.274) were independent risk factors for BM in HER2-positive BC patients. Prognostic data of all 65 HER2-positive BC patients with BM showed that the time from the diagnosis of BC to the development of breast cancer brain metastasis (BCBM) was 36.3 months (95% CI: 30.0 to 42.1 months). The time from the diagnosis of first recurrence and metastasis stage to the diagnosis of BCBM was 11.35 months (95% CI: 7.1 to 18.4 months). The time from the diagnosis of BCBM to the time of follow-up was 24.1 months (95% CI: 13.9 to 37.5 months). Up until the time of follow-up data, a total of 38 patients had died, and the time from the diagnosis of BM of these 38 patients to death was 11.0 months (95% CI: 9.0 to 20.4 months). Conclusion: The prognosis of HER2-positive BC patients with BM was poor due to the lack of effective treatments for BM. Age ≤40 years and first metastatic site with lung metastasis were the independent risk factors for BM in HER2-positive BC patients. Future research about pre-emptive medical interventions may help to improve the prognosis of HER2-positive BC patients with high risk to develop BM.

Overexpression of c-Ski promotes cell proliferation, invasion and migration of gastric cancer associated fibroblasts.

The present study aimed to investigate the effects of c-Ski on cell proliferation, invasion and migration of gastric cancer associated fibroblasts (CAFs). Expression of c-Ski in gastric cancer (GC) tissues was determined using immunohistochemistry. Both CAFs and non-cancerous gastric fibroblasts (NGFs) were isolated and cultured. c-Ski and Smad3 were over-expressed or knocked down using pcDNA3.0-c-Ski/Smad3 or siRNA, respectively. Cell viability, invasion and migration were measured and expression of c-Ski, α-SMA, and Smad3 in cells was determined using real time quantitative PCR (RT-qPCR) and Western blotting. Expression of c-Ski was significantly higher in both in GC tissues and cell lines, and was the highest in tissues of diffuse type. Both c-Ski and α-SMA were significantly over-expressed in CAFs compared with that in the NGFs. When c-Ski was over-expressed in NGFs, cell viability, cell invasion and migration were all enhanced and expression of Smad3 was downregulated. When c-Ski was inhibited, cell viability, cell invasion, and migration were all suppressed and expression of Smad3 was upregulated. Meanwhile, overexpression of Smad3 significantly reversed the effects of over-expressed c-Ski in NGFs, and knockdown of Smad3 dramatically reversed the effects of si-c-Ski in CAFs. Over-expressed c-Ski could enhance cell viability, promote cell invasion, and migration of GC CAFs, and the effects might be through regulation of Smad3 signaling. This study may give deeper insights for relationship between c-Ski and CAFs, as well as role of c-Ski in cancer development, and also provide some novel research targets for treatment of GC.

Association of postlunch napping duration and night-time sleep duration with cognitive impairment in Chinese elderly: a cross-sectional study.

OBJECTIVE: To analyse the independent and combined associations of postlunch napping duration and night-time sleep duration with risk of cognitive impairment among Chinese elderly. DESIGN: A cross-sectional study. SETTING: We analysed the data from Zhejiang Ageing and Health Cohort, a population-based survey of seven counties located in Zhejiang province in eastern China. PARTICIPANTS: 10 740 participants aged 60 years or older were included in final analysis. PRIMARY AND SECONDARY OUTCOME MEASURES: Cognitive impairment was assessed through Mini-Mental State Examination. Data on sleep-related characteristics was collected in the behavioural habits section within the questionnaire. RESULTS: Relative to participants with 1-30 min of postlunch napping, those who did not nap and who napped longer had significantly higher risks for cognitive impairment. OR of cognitive impairment were 1.41 (95% CI 1.14 to 1.75) for participants with longer night-time sleep duration (≥9 hours), compared with those sleeping 7-8.9 hours. In addition, combined effects were further identified. Participants with both longer night-time sleep duration (≥9 hours) and longer postlunch napping duration (>60 min) (OR=2.01, 95% CI 1.30 to 3.13), as well as those with both longer night-time sleep duration (≥9 hours) and appropriate postlunch napping duration (1-30 min) (OR=2.01, 95% CI 1.20 to 3.38), showed significantly higher risk of cognitive impairment than those with sleeping 7-8 hours and napping 1-30 min. Meanwhile, a 34% increase in odds of cognitive impairment was observed in participants with both shorter night-time sleep duration (5-6.9 hours) and no napping. CONCLUSION: Both postlunch napping duration and night-time sleep duration were independently and jointly associated with cognitive impairment, which needs verification in prospective studies.

[Effects of Huaiqihuang granules on immune function in children with severe Mycoplasma pneumoniae pneumonia].

OBJECTIVE: To observe the effects of Huaiqihuang granules on the immune function in children with severe Mycoplasma pneumoniae pneumonia. METHODS: Pediatric inpatients with severe Mycoplasma pneumoniae pneumonia were randomly divided into Huaiqihuang granule treatment group (n=51) and conventional treatment group (n=47). The Huaiqihuang granule treatment group was orally administered Huaiqihuang granules in addition to the conventional treatment, while the conventional treatment group received conventional treatment only. Levels of serum IgA, IgG, and IgM, percentages of CD4+ and CD8+ T lymphocyte subsets, and CD4+/CD8+ ratio were examined in the two groups. The incidence rate of respiratory tract re-infection within three months following treatment was compared between the two groups. RESULTS: The levels of serum IgA, IgG, and IgM, the percentage of CD4+ T lymphocytes, and the CD4+/CD8+ ratio were significantly higher in the Huaiqihuang granule treatment group than in the conventional treatment group three months after treatment (P<0.05). In contrast, the percentage of CD8+ T lymphocytes was significantly lower in the Huaiqihuang granule treatment group than in the conventional treatment group (P<0.05). In addition, the incidence rate of respiratory tract re-infection within three months following treatment was significantly lower in the Huaiqihuang granule treatment group than in the conventional treatment group (P<0.05). CONCLUSIONS: Huaiqihuang granules can regulate immune functions and reduce the incidence of short-term respiratory tract re-infection in children with severe Mycoplasma pneumoniae pneumonia.

Drug resistance mechanisms of Mycoplasma pneumoniae to macrolide antibiotics.

Throat swabs from children with suspected Mycoplasma pneumoniae (M. pneumoniae) infection were cultured for the presence of M. pneumoniae and its species specificity using the 16S rRNA gene. Seventy-six M. pneumoniae strains isolated from 580 swabs showed that 70 were erythromycin resistant with minimum inhibitory concentrations (MIC) around 32-512 mg/L. Fifty M. pneumoniae strains (46 resistant, 4 sensitive) were tested for sensitivity to tetracycline, ciprofloxacin, and gentamicin. Tetracycline and ciprofloxacin had some effect, and gentamicin had an effect on the majority of M. pneumoniae strains. Domains II and V of the 23S rRNA gene and the ribosomal protein L4 and L22 genes, both of which are considered to be associated with macrolide resistance, were sequenced and the sequences were compared with the corresponding sequences in M129 registered with NCBI and the FH strain. The 70 resistant strains all showed a 2063 or 2064 site mutation in domain V of the 23S rRNA but no mutations in domain II. Site mutations of L4 or L22 can be observed in either resistant or sensitive strains, although it is not known whether this is associated with drug resistance.

[Expression of MDR1 gene in cancer stem cells in breast cancer tissues of different molecular subtypes].

OBJECTIVE: To investigate the association between MDR1 gene expression in breast cancer stem cells and the molecular subtypes of breast cancer tissue. METHODS: According to ER, PR, Her-2 and CK5/14 expression profiles, 153 breast cancer specimens were divided into 5 molecular molecular subtypes, in which the expression of MDR1 was detected to analyze the relationship between MDR1 gene expression and the subtypes of breast cancer stem cells. RESULTS: The expression of MDR1 in Luminal A subtype breast cancer was 0.26∓0.04, which showed no significant difference from that of Luminal B subtype (0.31∓0.03, P>0.05). Compared with these two subtypes, HER-2 (+) subtype breast cancer tissues showed a significantly higher MDR1 expression(0.56∓0.05, P<0.05). MDR1 expression in Basal-like subtype and Normal-like subtype breast cancers was comparable (0.98∓0.01 vs 0.90∓0.15, P<0.05), but both significantly higher than that in Luminal A and B subtypes and HER-2 (+) subtype (P<0.05). CONCLUSION: The expression of MDR1 gene in cancer stem cells is related with the molecular subtypes of breast cancer tissue.

Prognostic value of different amounts of cancer stem cells in different molecular subtypes of breast cancer.

OBJECTIVE: To investigate the differences in the amount of cancer stem cells in different breast cancer subtypes. METHODS: THE SPECIMENS OF BREAST CANCER TISSUES WERE DIVIDED INTO FIVE GROUPS: group A (luminal A type), B (luminal B type), C (HER-2 + type), D (basal-like type) and E (normal-like type) according to their molecular subtypes. The cancer stem cell spheres in each group were measured through colony formation method, and then the relations of the amount of cancer stem cells with the subtypes of breast cancer were analyzed. RESULTS: The numbers of cancer stem cell spheres in group A and B were (1.1±0.2)/1,000 cells and (1.3±0.1)/1,000 cells, respectively, and this had no significant difference (P>0.05). The number of cancer stem cell spheres in group C was (8.6±1.0)/1,000 cells, which was significantly higher than that in group A and B (both P<0.05). The numbers of cancer stem cell spheres in group D and E were (22.4±1.2)/1,000 cells and (17.7±2.0)/1,000 cells, respectively, and this was statistically similar (P>0.05) but either was significantly higher than that in the other three groups (all P<0.05). CONCLUSIONS: Breast cancer tissues of different molecular subtypes contain varying amounts of cancer stem cells, and this may have certain significance for predicting the prognosis of breast cancer.

Molecular mechanisms of macrolide resistance in clinical isolates of Mycoplasma pneumoniae from China.

Fifty clinical Mycoplasma pneumoniae strains were isolated from 370 children with respiratory tract infections. Four strains were susceptible to macrolides, while the other 46 (92%) were macrolide resistant. The molecular mechanism of resistance was shown to be associated with point mutations in 23S rRNA at positions 2063 and 2064.