Objective:To investigate genetic variation profiles of δ-globin (HBD gene) and hematological phenotypes in Guangdong population.Methods:Retrospective case analysis was performed in this study. Blood samples of 11 616 couples who participated in free thalassemia screening in Guangzhou from July 2020 to December 2022 were collected which underwent blood routine tests and hemoglobin (Hb) capillary electrophoresis. According to the results, 154 samples were enrolled in this study: (1)group of 35 cases with HbA 2 <2.0% but no HbF band; (2)group of 64 cases with HbA 2 < 2.0% and HbF band; (3)group of 25 cases with HbA 2 <2.0% and suspected HbA 2 variants; (4) group of 25 cases with HbA 2 ≥2.0% and <3.5% and HbF band, as well as abnormal blood routine report [mean corpuscular volume (MCV) <82 fl and/or mean corpuscular hemoglobin (MCH) <27 pg]; (5)group of 5 cases with HbA 2 ≥2.0% and <3.0% accompanied with β thalassemia gene carriers Sanger sequencing was used to detect single nucleotide variants of δ-globin. Results:(1) A total of 22 genetic variations were detected, including 6 de novo variations, and the top 3 genetic variations were respectively c.-127T>C (57.02%, 65/114), c.-80T>C (9.65%, 11/114), c.349C>T (7.89%, 9/114). (2) In group of patients with HbA 2 <2.0% but no HbF band, 22 cases (62.85%, 22/35) had HBD gene variation, including 7 cases with MCV and MCH lower than reference values, 4 cases with α thalassemia; 13 cases had no HBD gene variation, including 12 cases with lower MCV and MCH. Among 19 cases with abnormal blood routine test results, levels of HbA 2 in patients (7 cases) with HBD gene variation were lower compared with those without HBD gene variation (12 cases) ( P<0.01%). (3)In group of patients with HbA 2<2.0% with HbF band, 59 cases (92.18%, 59/64) had HBD gene variations whose mutations all occurred in promoter region, and the HbF were all lower than 5.0%; 5 cases with HbF >5.0% had no HBD gene variation. (4) In group of patients with HbA 2 <2.0% and suspected HbA 2 variants, the detection rate was 100% (25/25) and δ-globin variants <1.0%. (5) In group of patients with HbA 2 ≥2.0% and <3.5% and HbF band accompanied with abnormal blood routine results, no HBD gene variation was found. (6) In group of 5 patients with HbA 2 ≥2.0% and <3.0% with β thalassemia gene carriers, HBD gene variation were found in all cases, and the level of HbA 2 was (2.62±0.17)% and HbF was (3.62±2.22)%. Conclusions:There are various genotypes of HBD gene variation, among which HBD: c.-127T>C is the most common in Guangdong population in China. Mutations in the promoter region may cause decrease in HbA 2 and increase in HbF which is mostly less than 5% but exceeds 5.0% when combined with β thalassemia. Our study enriched the gene mutation profiles of HBD gene in Guangdong population.

OBJECTIVE: The prepare decellularized extracellular matrix (ECM) scaffold materials derived from human cervical carcinoma tissues for 3D culture of cervical carcinoma cells. METHODS: Fresh human cervical carcinoma tissues were treated with sodium lauryl ether sulfate (SLES) solution to prepare decellularized ECM scaffolds. The scaffolds were examined for ECM microstructure and residual contents of key ECM components (collagen, glycosaminoglycan, and elastin) and genetic materials by pathological staining and biochemical content analysis. In vitro 3D culture models were established by injecting cultured cervical cancer cells into the prepared ECM scaffolds. The cells in the recellularized scaffolds were compared with those in a conventional 2D culture system for cell behaviors including migration, proliferation and epithelial-mesenchymal transition (EMT) wsing HE staining, immunohistochemical staining and molecular biological technology analysis. Resistance to 5-fluorouracil (5-Fu) of the cells in the two culture systems was tested by analyzing the cell apoptosis rates via flow cytometry. RESULTS: SLES treatment effectively removed cells and genetic materials from human cervical carcinoma tissues but well preserved the microenvironment structure and biological activity of ECM. Compared with the 2D culture system, the 3D culture models significantly promoted proliferation, migration, EMT and 5-Fu resistance of human cervical cancer cells. CONCLUSION The decellularized ECM scaffolds prepared using human cervical carcinoma tissues provide the basis for construction of in vitro 3D culture models for human cervical cancer cells.
Female ; Humans ; Decellularized Extracellular Matrix ; Extracellular Matrix ; Uterine Cervical Neoplasms ; Tissue Scaffolds/chemistry* ; Carcinoma ; Fluorouracil/pharmacology* ; Tissue Engineering ; Tumor Microenvironment

Sulforaphane (SFN), a natural anti-tumor compound from cruciferous vegetables, has been reported to induce protective autophagy to cancer cells, which might impair the anti-tumor efficiency of SFN. However, the accurate function and mechanism of SFN inducing autophagy in cancers are still obscure, especially in esophageal squamous cell carcinoma (ESCC), one of malignancies with high incidence in North China. Here, we mainly explored the potential function of autophagy upon SFN treatment in ESCC and molecular mechanism. We demonstrated that SFN could inhibit cell proliferation and induce apoptosis by activating caspase pathway. Moreover, we found activation of NRF2 pathway by SFN was responsible for the induction of autophagy and also a disadvantage element to the anti-tumor effects of SFN on ESCC, indicating that SFN might induce protective autophagy in ESCC. We, therefore, investigated effects of autophagy inhibition on sensitivity of ESCC cells to SFN and found that chloroquine (CQ) could neutralize the activation of SFN on NRF2 and enhance the activation of SFN on caspase pathway, thus improved the anti-tumor efficiency of SFN on ESCC

Objective:To explore the status quo and influencing factors of caregivers' perception of nurses' empowerment behavior in children with type 1 diabetes mellitus (T1DM) , and to provide scientific basis for focusing on and efficiently empowering caregivers of children with T1DM.Methods:This study was a cross-sectional survey. A total of 360 children with T1DM who were admitted to 2 tertiary hospitals in Zhejiang Province from January 2019 to May 2020 were selected by the convenient sampling method. General information questionnaire, Patient Perceptions of Patient-Empowering Nurse Behaviors Scale (PPPNBS) and General Self-Efficacy Scale (GSES) were used to investigate and analyze the influencing factors. A total of 360 questionnaires were issued and 347 valid questionnaires were recovered, with an effective response rate of 96.4%.Results:The PPPNBS score of caregivers of 347 children with T1DM was (185.85±50.05) . Multiple linear regression analysis showed that the course of the disease, the age of the caregiver, the education background of caregivers and self-efficacy sense were influencing factors of perception of empowerment behavior of caregivers of children with T1DM ( P<0.05) . Conclusions:The perception of caregivers of T1DM children with nurses' empowerment behavior is at a moderate level. The course of the disease, age of caregivers, the education background of caregivers and self-efficacy are the influencing factors in perception of caregivers' empowerment behaviors of T1DM children. Medical staff should formulate targeted intervention measures based on the influencing factors of the perception of empowerment behaviors to improve the perception of nurses' empowerment behaviors by caregivers of patients.

Objective:To study the expression level of CD73 in cutaneous melanoma and its clinical significance, and to explore its role and molecular mechanism in the occurrence and development of melanoma.Methods:Expression level of the gene NT5E in melanoma was analyzed according to RNA sequencing data from the CCLE database. CD73 expression level was tested in 145 melanoma tissues and corresponding para-tumor tissues by immunohistochemistry tissue microarray. The samples came from 78 male and 67 female patients who received surgical resection and were pathologically diagnosed as melanoma in the department of plastic surgery in Zhongshan Hospital from January of 2008 to December of 2017. The relationship between CD73 expression level and clinicopathological features as well as prognosis was analyzed. Melanoma cell line was transfected with CD73 gene overexpression lentivirus and negative control vectors, respectively. The transfection efficiency was verified by qRT-PCR and Western blot. CCK-8, wound healing and Transwell assays were used to evaluate the proliferation, migration and invasion abilities of the overexpression group and control group. Subcutaneous xenograft tumor models were established in nude mice to assess the tumorigenicity of the two groups. Transcriptome sequencing, GO and KEGG analyses were performed. The expression levels of pAKT and pGSK3β were detected by Western blot. The measurement data were expressed as Mean ±SD. Independent sample t-test was used for comparison between the two groups, and One-way ANOVA was employed for multi-group comparisons. The chi-square test was used for the categorical variables. P<0.05 was considered as statistically significant difference. Results:The expression of CD73 in melanoma was higher than that in para-tumor tissues ( P< 0.05). Higher expression level of CD73 was associated with higher Clark grade ( P= 0.014) and clinical stage ( P= 0.040). The overall survival and disease-free survival of patients with high-CD73-expression were significantly lower than those of patients with low-CD73-expression. CD73 expression, Clark grade and clinical stage are independent risk factors for the poor prognosis of patients with melanoma. Overexpression of CD73 significantly enhanced the proliferation, migration and invasion of melanoma cells in vitro and the tumorigenicity in vivo( P< 0.05). Transcriptome sequencing showed that CD73 was involved in several tumor related signaling pathways, among which the expression of pAKT and pGSK3β was elevated( P< 0.05). Conclusions:CD73 promotes the proliferation, migration and invasion of melanoma cells in vitro and the tumorigenicity in vivo through activating AKT signaling. CD73 is expected to become a new prognostic biomarker and therapeutic target for melanoma.

Objective:To study the expression level of CD73 in cutaneous melanoma and its clinical significance, and to explore its role and molecular mechanism in the occurrence and development of melanoma.Methods:Expression level of the gene NT5E in melanoma was analyzed according to RNA sequencing data from the CCLE database. CD73 expression level was tested in 145 melanoma tissues and corresponding para-tumor tissues by immunohistochemistry tissue microarray. The samples came from 78 male and 67 female patients who received surgical resection and were pathologically diagnosed as melanoma in the department of plastic surgery in Zhongshan Hospital from January of 2008 to December of 2017. The relationship between CD73 expression level and clinicopathological features as well as prognosis was analyzed. Melanoma cell line was transfected with CD73 gene overexpression lentivirus and negative control vectors, respectively. The transfection efficiency was verified by qRT-PCR and Western blot. CCK-8, wound healing and Transwell assays were used to evaluate the proliferation, migration and invasion abilities of the overexpression group and control group. Subcutaneous xenograft tumor models were established in nude mice to assess the tumorigenicity of the two groups. Transcriptome sequencing, GO and KEGG analyses were performed. The expression levels of pAKT and pGSK3β were detected by Western blot. The measurement data were expressed as Mean ±SD. Independent sample t-test was used for comparison between the two groups, and One-way ANOVA was employed for multi-group comparisons. The chi-square test was used for the categorical variables. P<0.05 was considered as statistically significant difference. Results:The expression of CD73 in melanoma was higher than that in para-tumor tissues ( P< 0.05). Higher expression level of CD73 was associated with higher Clark grade ( P= 0.014) and clinical stage ( P= 0.040). The overall survival and disease-free survival of patients with high-CD73-expression were significantly lower than those of patients with low-CD73-expression. CD73 expression, Clark grade and clinical stage are independent risk factors for the poor prognosis of patients with melanoma. Overexpression of CD73 significantly enhanced the proliferation, migration and invasion of melanoma cells in vitro and the tumorigenicity in vivo( P< 0.05). Transcriptome sequencing showed that CD73 was involved in several tumor related signaling pathways, among which the expression of pAKT and pGSK3β was elevated( P< 0.05). Conclusions:CD73 promotes the proliferation, migration and invasion of melanoma cells in vitro and the tumorigenicity in vivo through activating AKT signaling. CD73 is expected to become a new prognostic biomarker and therapeutic target for melanoma.

Objective:To investigate the risk factors for 90 d death after endovascular mechanical thrombectomy (MT) in patients with acute anterior circulation large-artery occlusive stroke.Methods:From October 2015 to March 2018, patients with acute anterior circulation large-artery occlusive stroke treated with MT in People's Hospital of Shanghai Pudong New Area and the Second Affiliated Hospital of Soochow University were enrolled retrospectively. The primary outcome events were defined as death within 90 d after operation. Univariate and multivariate logistic regression models were used to identify the independent risk factors for death within 90 d after operation. Results:A total of 116 patients were enrolled, 23 (19.8%) of them died within 90 d after operation. Univariate analysis showed that there were significant differences in age, baseline National Institutes of Health Stroke Scale (NIHSS) score, the Alberta Stroke Program Early CT Score (ASPECTS), and the proportion of the baseline NIHSS score classification (≤8, 9-15, ≥16), ASPECTS ≤7, the number of attempts to pass >3 times, modified Thrombolysis in Cerebral Infarction (mTICI) blood flow grade 2b/3, hemorrhagic transformation (HT), and symptomatic HT in the death group compared with the survival group (all P<0.05). Multivariate analysis showed that after adjusting for age, fasting blood glucose, baseline NIHSS score, number of attempts to pass >3, and mTICI grade 2b/3, lower ASPECTS (odds ratio [ OR] 0.647, 95% confidence interval [ CI] 0.456-0.917; P=0.014), longer time from onset to vascular recanalization ( OR 1.004, 95% CI 1.000-1.007; P=0.046) and symptomatic HT ( OR 13.522, 95% CI 2.719-67.258; P=0.001) were the independent predictors of death within 90 d. Conclusion:The ASPECTS, time from onset to recanalization, and symptomatic HT were the independent risk factors for death within 90 d after MT in patients with acute anterior circulation large-artery occlusive stroke.

Dysregulation of mTORC1/mTORC2 pathway is observed in many cancers and mTORC1 inhibitors have been used clinically in many tumor types; however, the mechanism of mTORC2 in tumorigenesis is still obscure. Here, we mainly explored the potential role of mTORC2 in esophageal squamous cell carcinoma (ESCC) and its effects on the sensitivity of cells to mTOR inhibitors. We demonstrated that RICTOR, the key factor of mTORC2, and p-AKT (Ser473) were excessively activated in ESCC and their overexpression is related to lymph node metastasis and the tumor-node-metastasis (TNM) phase of ESCC patients. Furthermore, we found that mTORC1/ mTORC2 inhibitor PP242 exhibited more efficacious anti-proliferative effect on ESCC cells than mTORC1 inhibitor RAD001 due to RAD001-triggered feedback activation of AKT signal. Another, we demonstrated that down-regulating expression of RICTOR in ECa109 and EC9706 cells inhibited proliferation and migration as well as induced cell cycle arrest and apoptosis. Noteworthy, knocking-down stably RICTOR significantly suppresses RAD001-induced feedback activation of AKT/PRAS40 signaling, and enhances inhibition efficacy of PP242 on the phosphorylation of AKT and PRAS40, thus potentiates the antitumor effect of RAD001 and PP242 both and . Our findings highlight that selective targeting mTORC2 could be a promising therapeutic strategy for future treatment of ESCC.

Objective To comparatively analyze the safety and efficacy of direct mechanical thrombectomy and bridging therapy for patients with acute anterior circulation large-artery occlusive stroke.Methods A total of 116 patients with acute anterior circulation large-artery occlusive stroke,admitted to our hospitals from October 2015 to March 2018,were chosen in our study;their clinical data were analyzed retrospectively.Among them,63 patients accepted direct mechanical thrombectomy and 53 accepted bridging therapy.The preoperative baseline data and the diagnoses and treatments of the two groups were analyzed;the degrees of modified thrombolysis in cerebral infarction (mTICI),incidences of hemorrhage transformation and symptomatic intracranial hemorrhage,and modified Rankin scale (mRS) scores and mortality rate 90 d after operation were compared between the two groups.Results The preoperative Alberta stroke program early CT scale (ASPECTS) and Glasgow Coma Scale (GCS) scores of the direct mechanical thrombectomy group were significantly lower than those of the bridge therapy group (P＜0.05),and the time from onset to admission was significantly longer than that of the bridging therapy group (P＜0.05).The incidence of postoperative hemorrhage transformation in the direct mechanical thrombectomy group was significantly higher than that in the bridging therapy group (34.9％ vs.17.0％,P＜0.05),but there were no significant differences in the effective recanalization rate (69.8％ vs.79.3％),intracranial symptomatic hemorrhage rate (15.9％ vs.7.6％),favorable outcome rate (28.6％ vs.35.9％) and mortality (22.2％ vs.17.0％) between the two groups (P＞0.05).Conclusion The clinical efficacy and safety of direct mechanical thrombectomy and bridging therapy for patients with acute anterior circulation large-artery occlusive stroke are similar.

Objective:To explore the influencing factors for intracranial hemorrhage (ICH) in patients with acute large vessel occlusion stroke (ALVOS) of anterior circulation after mechanical thrombectomy (MT).Methods:From October 2015 to March 2018, the clinical data of 116 patients with ALVOS of anterior circulation accepted MT were analyzed retrospectively in our hospitals. These patients were divided into ICH group ( n=31) and non-ICH group ( n=85) according to whether there was ICH after operation. Univariate and multivariate Logistic regression models were used to analyze the independent influencing factors for ICH after MT. Results:Univariate analysis showed that there were significant differences in preoperative Alberta Stroke Program Early CT (ASPECT) scores, baseline systolic pressure, and preoperative blood glucose level between ICH group and non-ICH group ( P<0.05). Multivariate analysis showed that after adjusting baseline Glasgow coma scale scores, preoperative ASPECT scores ( OR=0.770, 95%CI: 0.610-0.0.971, P=0.027), baseline systolic pressure ( OR=1.029, 95%CI: 1.005-1.054, P=0.017), preoperative blood glucose level ( OR=1.177, 95%CI: 1.010-1.372, P=0.036) were independent influencing factors for ICH. Conclusion:The patients with low preoperative ASPECT scores, high baseline systolic pressure, and high blood glucose are prone to have ICH after MT in patients with ALVOS of anterior circulation.