Development and validation of a nomogram for predicting survival of breast cancer patients with ipsilateral supraclavicular lymph node metastasis.

BACKGROUND: Breast cancer patients with ipsilateral supraclavicular lymph node metastasis (ISLNM) but without distant metastasis are considered to have a poor prognosis. This study aimed to develop a nomogram to predict the overall survival (OS) of breast cancer patients with ISLNM but without distant metastasis. METHODS: Medical records of breast cancer patients who received surgical treatment at the Affiliated Cancer Hospital of Zhengzhou University, Jiyuan People's Hospital and Huaxian People's Hospital between December 21, 2012 and June 30, 2020 were reviewed retrospectively. Overall, 345 patients with pathologically confirmed ISLNM and without evidence of distant metastasis were identified. They were further randomized 2:1 and divided into training (n = 231) and validation (n = 114) cohorts. A nomogram to predict the probability of OS was constructed based on clinicopathologic variables identified by the univariable and multivariable analyses. The predictive accuracy and discriminative ability were measured by calibration plots, concordance index (C-index), and risk group stratification. RESULTS: Univariable analysis showed that estrogen receptor-positive (ER+), progesterone receptor-positive (PR+), human epidermal growth factor receptor 2-positive (HER2+) with Herceptin treatment, and a low axillary lymph node ratio (ALNR) were prognostic factors for better OS. PR+, HER2+ with Herceptin treatment, and a low ALNR remained independent prognostic factors for better OS on multivariable analysis. These variables were incorporated into a nomogram to predict the 1-, 3-, and 5-year OS of breast cancer patients with ISLNM. The C-indexes of the nomogram were 0.737 (95% confidence interval [CI]: 0.660-0.813) and 0.759 (95% CI: 0.636-0.881) for the training and the validation cohorts, respectively. The calibration plots presented excellent agreement between the nomogram prediction and actual observation for 3 and 5 years, but not 1 year, OS in both the cohorts. The nomogram was also able to stratify patients into different risk groups. CONCLUSIONS: In this study, we established and validated a novel nomogram for predicting survival of patients with ISLNM. This nomogram may, to some extent, allow clinicians to more accurately estimate prognosis and to make personalized therapeutic decisions for individual patients with ISLNM.

[Cloning, expression and purification of Schistosoma japonicum ribosomal protein S4 as well as the preliminary study of the diagnostic value of the recombinant protein].

OBJECTIVE: To express and purify Schistosoma japonicum ribosomal protein S4(SjRPS4) in Escherichia coli, and assess its value in immunodiagnosis of Schistosomiasis japonica. METHODS: Gene fragment of SjRPS4 was amplified by screening the cercaria cDNA library of Schistosoma japonicum. The target gene was cloned into the expressive vector pQE30 and transformed into E. coli M15. The recombinant protein expression was induced by isopropylthio-ß-D-galactoside (IPTG). This fusion protein was purified by Ni(2+)-NTA chromatography and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blot and ELISA. RESULTS: The plasmid pQE30/SjRPS4 was constructed successfully and expressed a SjRPS4 fusion protein in E. coli as showing a single special band on SDS-PAGE gel at Mr 30 × 10(3) position. It reached a purity of above 90% after purification. The Western blot result confirmed that the recombinant protein could specifically react with the serum samples from patients of schistosomiasis. Detecting the serum of Schistosomiasis japonica patients by ELISA, the sensitivity and specificity of the ELISA method were 90.91% (70/77) and 92.59% (25/27), the positive rate of recombinant protein expression was 67.30% (70/104). There was no cross-reaction with paragonimiasis patients' serum. CONCLUSION: Protein SjRPS4 was successfully cloned and expressed, and it was confirmed that SjRPS4 antibodies were valuable in the diagnosis of Schistosomiasis japonica.

Schistosoma japonicum: screening of cercariae cDNA library by specific single-chain antibody against SIEA26-28 ku and immunization experiment of the recombinant plasmids containing the selected genes.

To obtain the gene encoding SIEA26-28 ku, which has been proven to be a potential anti-schistosomiasis vaccine candidate, screening Schistosoma japonicum (Sj) cercariae cDNA library with soluble specific single-chain antibody (SIEA26-28 ku-scFv) was performed. A large amount of specific single-chain antibody was harvested through construction of recombinant expression vector pET32a/scFv. The protein was purified and characterized. By using this protein (PET32a-scFv) as a probe, S. japonicum cercariae cDNA library was screened. Two strong positive clones were selected, and their eukaryotic recombinant plasmids were constructed. These genes were named as S. japonicum ribosomal protein S4 (SjRPS4) and S. japonicum ribosomal protein L7 (SjRPL7), respectively. Experiments of mice showed that both SjRPS4 and SjRPL7 DNA vaccines could induce significant immunoprotection. Result of these experiments further proved that the specific single-chain antibody is a very valuable tool in screening of cDNA library to get the corresponding molecules.

[Research progress on schistosome vaccine and prospect of its application in China].

Development of vaccine against schistosomiasis japonica has been incorporated into WHO/TDR and China's main disease control research programs. In recent years,the research on the anti-schistosomiasis vaccine has made significant progress. With the development of proteomics and molecular biology technology, Anti-Schistosoma japonicum vaccine research has been developed to a stage of genetic engineering in our country and DNA vaccines have become the main direction. It reveals new ways to enhance the immunoprotection of Schistosoma japonicum vaccine through screening new candidate antigens, optimizing combination of the mixed/multivalent vaccines, or adjuvant addition.