RNAi is a process of effective gene silencing. Human papillomaviruses 16 (HPV16), as the most prevalent genotype of cervical cancer, encodes the E6 and E7 oncogenes, which is pivotal for maintenance of malignant phenotypes. Recently, some significant advances have been obtained in RNA interfering effect on HPV16 E6/E7. In vitro, it has been developed more effective vector systems and screened for more specific targeted sequences. In vivo, the corresponding animal models have been established, which lay a good foundation for the clinical research.

Objective To investigate the application of contrast-enhanced ultrasonograhy CEUS in the early assessment of antiangiogenic treatment in metastatic colorectal cancer Methods Among 41 colorectal cancer patients with liver metastases enrolled in this study 25 patients received treatment of antiangiogenic treatment bevacizumab combined with chemotherapy BV group and 16 patients received chemotherapy only non-BV group To assess the treatment responses in the liver lesions between and within these two groups CEUS scans were performed before the treatment T0 or??baseline on the day 2 T1 and on the day 7 T2 after the treatment The percentage differences were evaluated according to the CEUS quantitative parameters such as rise time RT peak intensity PI mean transit time MTT area under the curve AUC time from peak to one half TPH wash in slope WIS and time to peak TTP Results On T1 both the RT and MTT increased within the BV group and decrease within the non-BV group when compared with their own baselines the differences were statistically significant P <0 05 On T2 the MTT increased within the BV group and decreased within the non-BV group when compared with their baselines the differences again were statistically significant P <0 05 All other parameters measured on T1 and T2 did not show a significant difference when compared with baselines Within the BV group the RT decreased and WIS and TPH increased for the responding subjects when compared with the non-responding subjects the differences were statistically significant P < 0 05 Within the non-BV group however no statistically significant differences were found for all parameters between the responding subjects and non-responding subjects P >0 05 Conclusions CEUS can be used to assess the treatment response in liver metastatic patients with colorectal cancer by monitoring the change of the tumor perfusion before and after the treatment.

Objective To investigate the effects of contrast dose and region of interest (ROI) depth on quantitative analysis of liver by contrast-enhanced ultrasound (CEUS) during clinical application.Methods After bolus injection of contrast agent,the change of quantitative parameters [including echo mean(EM),rise time(RT),peak intensity(PI),mean transit time(MTT),area under the curve(AUC),time from peak to one half(TPH),wash in slope(WIS),time to peak(TTP)] of time-intensity curves were analyzed based on groups from different doses (1.0 ml and 1.6 ml) and different depth (＜30 mm,30-60 mm,and ≥60 mm).Results MTT and TPH were increased with dose increasing from 1.0 ml to 1.6 ml (P＜0.05).With the dose 1.0 ml,TPH,WIS,PI,AUC and MTT showed significant difference when the depth of the ROI changes (P ＜0.05),with the depth increased,TPH,WIS,PI,and AUC all decreases and MTT increases.For all the other parameters,no significant changes were found (P ＞0.05).Conclusions CEUS and its imaging process can directly influence the accuracy of the parameters from the quantitative analysis.Standardization of contrast agent with predefined dose and depth can potentially facilitate future clinical studies in liver CEUS.

Objective:To investigate the value of ultrasound combined with thyroglobulin (Tg) in preoperative N staging of thyroid carcinoma.Methods:The clinical data of 1 138 patients with thyroid carcinoma in the Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital) from August 2018 to October 2020, who confirmed by surgery and pathology were analyzed retrospectively. The 1 138 cases were divided into pN0, pN1a, and pN1b stages. Kappa consistency test was used to analyze the consistency of ultrasound evaluation of N staging and pathological N staging. Pathology result was taken as the gold standard to analyze the correlation between some preoperative serum markers and lymph node metastasis. The ROC curve was used to compare the diagnostic value of ultrasound, Tg, and ultrasound combined with Tg for lymph node metastasis.Results:The preoperative ultrasound assessment of N staging was moderately consistent with pathology(Kappa=0.459, P<0.01). Between pN0 and pN1(pN1a+ pN1b) stages, the differences in free triiodothyronine (fT3), anti-thyroid peroxidase antibody (TPOAb) and Tg were statistically significant (all P<0.05). Among the different indicators, only Tg had significant effect on lymph node metastasis ( P<0.01) .The area under the ROC curve (AUC) of Tg in predicting lymph node metastasis of thyroid cancer was 0.679, while the best cut-off value for Tg was 25.245 μg/L. The AUC of only ultrasound and ultrasound combined with Tg were 0.699 and 0.775, respectively. Therefore, combined diagnosis method was better than ultrasound only. Conclusions:Preoperative ultrasound and thyroglobulin has a specific value in evaluating the N staging of thyroid carcinoma. The combination of the two is more valuable in the diagnosis of lymph node metastasis than ultrasound only.

Objective To explore the correlation of nerve function and prognosis with serum uric acid(UA),homocysteine(Hcy)and low-density lipoprotein cholesterol(LDL-C)in patients with acute cerebral infarction(ACI)after alteplase intravenous thrombolysis.Methods A total of 220 ACI patients undergoing thrombolysis in Changsha First Hospital ICU between January 2020 and December 2022 were enrolled,and according to mRS score at 3 months after thrombolysis,they were divided into poor prognosis group(mRS score＞2,91 cases)and good prognosis group(mRS score ≤2,129 cases).The serum levels of UA,Hcy and LDL-C were compared between the two groups.The correlation between the three indexes and score of National Institutes of Health Stroke Scale(NIHSS),and their predictive value for poor prognosis were analyzed.Results At 1 and 3 d after thrombolysis,the serum levels of UA,Hcy and LDL-C and NIHSS score were sig-nificantly decreased in both groups,and the serum levels of UA and Hcy and NIHSS score at 3 d after thrombolysis were significantly lower than those at 1 d(P＜0.05).The poor prognosis group had obviously higher serum levels of UA,Hcy and LDL-C and NIHSS score at 1 and 3 d after thrombolysis than the good prognosis group(P＜0.05,P＜0.01).Pearson correlation analysis showed that the serum levels of UA,Hcy and LDL-C were positively correlated with NIHSS score at 1 and 3 d after thrombolysis(P＜0.01).ROC curve analysis indicated that the AUC values of UA,Hcy and LDL-C at 1 d after thrombolysis for predicting poor prognosis were 0.707(95％CI:0.639-0.776),0.800(95％CI:0.739-0.860)and 0.624(95％CI:0.550-0.698),respectively,while the values of them at 3 d after thrombolysis were 0.655(95％CI:0.583-0.726),0.730(95％CI:0.664-0.795)and 0.573(95％CI:0.497-0.649),respectively.Conclusion In ACI patients after thrombolysis,the serum levels of UA,Hcy and LDL-C are increased in those with poor prognosis,and are associated with the severity of nerve injury.The levels at 1 d after throm-bolysis have good predictive value for poor prognosis.

Objective:To evaluate the left ventricular myocardial work parameters of the animal models with left bundle branch block (LBBB) and evaluate the effects of LBBB on left ventricular function and motion pattern by pressure-strain loops (PSL) of speckle tracking imaging (STI).Methods:In Twenty-four healthy male beagles, LBBB was induced by radio frequency ablation under anesthesia, and blood pressure was measured at the same time. Electrocardiograms and echocardiography images were acquired before (baseline), 30 minutes after (acute-LBBB) and 3 months after(chronic-LBBB) the creation of LBBB respectively. STI was applied to measure the left ventricular global longitudinal strain (GLS) and obtain the the PSL of each time point to evaluate the left ventricular global and segmental myocardial work parameters.Results:Compared to the baseline, the global work efficiencies(GWE) were obviously reduced ( P<0.05) and global wasted works(GWW) were significantly increased ( P<0.01) in the acute-LBBB and chronic-LBBB, significant differences were observed in GLS between acute-LBBB and baseline( P=0.04). In baseline, the work efficiency (WEsept) and the constructive work (CWsept) in the basal and middle segments of the septal wall were both obviously higher than the corresponding segments of left ventricular lateral wall( P<0.01), while the distribution of the wasted work(WWsept) was opposite( P<0.01). In acute-LBBB, the WEsept of all segments were significantly decreased ( P<0.05), the WWsept were obviously increased( P<0.05), the CWsept of basal segment was significantly reduced( P=0.01), while the wasted work in the basal segment of lateral wall(WWlat) was increased( P=0.04) compared with the baseline. Compared with the acute-LBBB, the WEsept of basal and middle segments were mildly recovery( P=0.03) in chronic-LBBB, but were still lower than the baseline ( P=0.001), the changes of the other myocardial work parameters of septal and lateral wall were similar to the acute-LBBB. Conclusions:Both acute-LBBB and chronic-LBBB can lead to the changes of left ventricular global and segmental myocardial work parameters. The myocardial work parameters of left ventricle can quantitatively analyze the changes of left ventricular function and motion pattern of the LBBB.

Objective:To analyze the long-term prognosis and prognostic factors of allergic bronchopulmonary aspergillosis(ABPA) in children suffering from cystic fibrosis (CF).Methods:An observational study was performed.All children who were admitted to the Department of Respiratory, Beijing Children′s Hospital, Capital Medical University from August 2014 to June 2018, with more than 2 years of followed up for the diagnosis of CF accompanied by ABPA were involved.Results:Three children met the inclusion criteria, with 2 boys and 1 girl, and their diagnostic age were 14, 8 and 9 years old, respectively.The follow-up duration ranged from 2 to 6 years.All the 3 cases were treated with systemic corticosteroids and antifungal agent.In case 1, the initial dose of prednisone was 0.75 mg/(kg·d), and the course of treatment was more than 5 years.The corticosteroid-dependent patient suffered from expectoration and chest pain, and radiographic findings indicated exacerbation, while his lung function was normal.Treating with initial dose of prednisone 2 mg/(kg·d) for 9 months, case 2 had normal serum immunoglobulin E(IgE) concentration, but his pulmonary artery was infiltrated by lesions, thus leasing to lobectomy.In case 3, the initial dose of prednisone was 0.6 mg/(kg·d), and the course of treatment was 18 months.And she developed persistent hypoxemia, and decreased pulmonary function, so lung transplantation was necessary 2 years after diagnosis.Conclusions:Systemic glucocorticoid combined with antifungal therapy is the main treatment for CF with ABPA, but there are individual differences in the efficacy.The level of serum total IgE is not always consistent with lung function and chest images.The overall prognosis is poor, and it is infeasible to evaluate the prognosis by single factor.

The widespread application of next generation sequencing (NGS) in clinical settings has enabled testing, diagnosis, treatment and prevention of genetic diseases. However, many issues have arisen in the meanwhile. One of the most pressing issues is the lack of standards for reporting genetic test results across different service providers. The First Forum on Standards and Specifications for Clinical Genetic Testing was held to address the issue in Shenzhen, China, on October 28, 2017. Participants, including geneticists, clinicians, and representatives of genetic testing service providers, discussed problems of clinical genetic testing services across in China and shared opinions on principles, challenges, and standards for reporting clinical genetic test results. Here we summarize expert opinions presented at the seminar and report the consensus, which will serve as a basis for the development of standards and guidelines for reporting of clinical genetic testing results, in order to promote the standardization and regulation of genetic testing services in China.

ObjectiveTo explore the influence of the core symptoms of attention deficit hyperactivity disorder （ADHD） on behavioral problems of children with ADHD propensity， so as to provide references for early identification and targeted intervention for children with ADHD propensity. MethodsFrom July to August 2021， 25 children with ADHD propensity were screened as the ADHD propensity group， and 25 children matched for age， gender and grade were included as the normal group in an elementary school in Guangzhou. ADHD core symptoms were assessed by the Chinese version of the Swanson Nolan and Pelham， version IV-parent form for ADHD （SNAP-IV）， and behavioral problems were assessed by Questionnaire-Children with Difficulties （QCD） and Conners Parental Symptom Questionnaire （PSQ）. Spearman correlation analysis was used to examine the correlation between ADHD core symptoms and QCD and PSQ scores， and hierarchical linear regression analysis was used to explore the effect of ADHD core symptoms on behavioral problems. Results① The differences between the groups showed that both attention deficit and hyperactivity-impulsivity factor scores were higher in the ADHD propensity group than those in the normal group （t=7.771， 6.726， P<0.01）. ② Correlation analysis showed that the attention deficit factor score was negatively correlated with QCD total score （r=-0.440， P<0.05）， and positively correlated with the learning problem factor score of PSQ （r=0.457， P<0.05）. The score of hyperactivity-impulsivity was negatively correlated with score of anxiety factor in PSQ （r=-0.457， P<0.05）， and positively correlated with impulse-hyperactivity factor score （r=0.552， P<0.01）. ③ Hierarchical linear regression analysis showed that the attention deficit factor score negatively predicted the total score of QCD （B=-0.682， P<0.05， R²=0.468）. The hyperactivity-impulsivity factor score had a negative predictive effect on the anxiety factor score of PSQ （B=-0.048， P<0.05， R²=0.367）， and had a positive predictive effect on the impulsivity-hyperactivity factor score （B=0.077， P<0.01， R²=0.424）. ConclusionChildren with ADHD propensity have significant attention deficit symptoms， hyperactivity-impulsivity symptoms and behavioral problems， and the attention deficit may be the main cause of their daily behavioral problems， while hyperactivity-impulsivity may be the main cause of their impulsive-hyperactivity problems.

BACKGROUND: Ischemic acute kidney injury (AKI) is a common syndrome associated with considerable mortality and healthcare costs. Up to now, the underlying pathogenesis of ischemic AKI remains incompletely understood, and specific strategies for early diagnosis and treatment of ischemic AKI are still lacking. Here, this study aimed to define the transcriptomic landscape of AKI patients through single-cell RNA sequencing (scRNA-seq) analysis in kidneys. METHODS: In this study, scRNA-seq technology was applied to kidneys from two ischemic AKI patients, and three human public scRNA-seq datasets were collected as controls. Differentially expressed genes (DEGs) and cell clusters of kidneys were determined. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, as well as the ligand-receptor interaction between cells, were performed. We also validated several DEGs expression in kidneys from human ischemic AKI and ischemia/reperfusion (I/R) injury induced AKI mice through immunohistochemistry staining. RESULTS: 15 distinct cell clusters were determined in kidney from subjects of ischemic AKI and control. The injured proximal tubules (PT) displayed a proapoptotic and proinflammatory phenotype. PT cells of ischemic AKI had up-regulation of novel pro-apoptotic genes including USP47 , RASSF4 , EBAG9 , IER3 , SASH1 , SEPTIN7 , and NUB1 , which have not been reported in ischemic AKI previously. Several hub genes were validated in kidneys from human AKI and renal I/R injury mice, respectively. Furthermore, PT highly expressed DEGs enriched in endoplasmic reticulum stress, autophagy, and retinoic acid-inducible gene I (RIG-I) signaling. DEGs overexpressed in other tubular cells were primarily enriched in nucleotide-binding and oligomerization domain (NOD)-like receptor signaling, estrogen signaling, interleukin (IL)-12 signaling, and IL-17 signaling. Overexpressed genes in kidney-resident immune cells including macrophages, natural killer T (NKT) cells, monocytes, and dendritic cells were associated with leukocyte activation, chemotaxis, cell adhesion, and complement activation. In addition, the ligand-receptor interactions analysis revealed prominent communications between macrophages and monocytes with other cells in the process of ischemic AKI. CONCLUSION Together, this study reveals distinct cell-specific transcriptomic atlas of kidney in ischemic AKI patients, altered signaling pathways, and potential cell-cell crosstalk in the development of AKI. These data reveal new insights into the pathogenesis and potential therapeutic strategies in ischemic AKI.
Humans ; Mice ; Animals ; Transcriptome/genetics* ; Ligands ; Kidney/metabolism* ; Acute Kidney Injury/metabolism* ; Ischemia/metabolism* ; Reperfusion Injury/metabolism* ; Sequence Analysis, RNA ; Adaptor Proteins, Signal Transducing/metabolism* ; Tumor Suppressor Proteins/metabolism*