ObjectiveTo analyze the epidemiological characteristics of a local dengue fever cluster outbreak in Qingpu District of Shanghai in 2024, and to provide a reference for subsequent dengue fever prevention and control. MethodsSeven confirmed local dengue fever cases reported through the National Notifiable Infectious Diseases Surveillance System in Qingpu District of Shanghai in 2024 were selected as the research subjects. Descriptive epidemiological methods were used to conduct investigation and analysis from the aspects of onset, medical treatment and reporting, clinical symptoms, travel and contact history within 15 days before onset, and activity trajectories. ResultsA total of 7 cases were identified in this outbreak. None of the cases had a travel history to dengue-endemic areas within 15 days prior to onset, while all had shared exposure environments and mosquito bite histories, indicating a local clustered transmission pattern. The main clinical manifestations included fever (100.00%) and myalgia (42.86%). All 7 cases were positive for dengue virus serotype 2 (DENV-2) by nucleic acid testing. Genetic sequencing showed that the virus strains belonged to the Cosmopolitan genotype and were most closely related to the epidemic DENV strains circulating in southern China in recent years. ConclusionThis outbreak might be a local secondary infection caused by the short-term stay of dengue fever-infected individuals, and the possible source of importation was dengue fever endemic areas in southern China.

L-citrulline is a nonprotein amino acid that plays an important role in human health and has great market demand. Although microbial cell factories have been widely used for biosynthesis, there are still challenges such as genetic instability and low efficiency in the biosynthesis of L-citrulline. In this study, an efficient, plasmid-free, non-inducible L-citrulline-producing strain of Escherichia coli BL21(DE3) was engineered by combined strategies. Firstly, a chassis strain capable of synthesizing L-citrulline was constructed by block of L-citrulline degradation and removal of feedback inhibition, with the L-citrulline titer of 0.43 g/L. Secondly, a push-pull-restrain strategy was employed to enhance the L-citrulline biosynthesis, which realized the L-citrulline titer of 6.0 g/L. Thirdly, the NADPH synthesis and L-citrulline transport were strengthened to promote the synthesis efficiency, which achieved the L-citrulline titer of 11.6 g/L. Finally, fed-batch fermentation was performed with the engineered strain in a 3 L fermenter, in which the L-citrulline titer reached 44.9 g/L. This study lays the foundation for the industrial production of L-citrulline and provides insights for the modification of other amino acid metabolic networks.
Citrulline/biosynthesis* ; Escherichia coli/genetics* ; Metabolic Engineering/methods* ; Fermentation ; NADP/biosynthesis*

The incidence of cancer continues to rise, and improving treatment outcomes and quality of life for cancer patients has become a major goal. Death anxiety is prevalent throughout the entire disease trajectory of cancer patients. It not only severely impairs their psychological well-being and quality of life but also undermines treatment confidence and rational decision-making. Therefore, it warrants significant clinical attention. This paper reviews the current status of assessment, influencing factors, and intervention strategies for death anxiety in cancer patients, aiming to provide a reference for healthcare professionals.

The incidence of cancer continues to rise, and improving treatment outcomes and quality of life for cancer patients has become a major goal. Death anxiety is prevalent throughout the entire disease trajectory of cancer patients. It not only severely impairs their psychological well-being and quality of life but also undermines treatment confidence and rational decision-making. Therefore, it warrants significant clinical attention. This paper reviews the current status of assessment, influencing factors, and intervention strategies for death anxiety in cancer patients, aiming to provide a reference for healthcare professionals.

The quality of medical insurance settlement lists directly affects the accuracy and rationality of medical insurance payments. This study analyzed the quality problems existing in L City′s 2023 medical insurance settlement list from three dimensions: integrity, standardization and rationality, and introduced a series of medical insurance settlement list quality management measures implemented with a problem-oriented approach. City L has effectively improved the overall quality of medical insurance settlement list by compiling local medical insurance settlement list filling standards, building a closed-loop model for quality control management of medical insurance settlement lists, establishing a quality grading management system for medical insurance settlement lists, and incorporating the quality of medical insurance settlement lists into performance appraisal and agreement management. The proportion of medical insurance settlement lists with quality problems in the city decreased from 8.12% (87 500 out of 1 077 300) in 2023 to 1.80% (20 700 out of 1 152 000) in 2024. The number of problems related to integrity, standardization, and rationality decreased from 39 900, 129 300, and 105 300 to 12 600, 14 800, and 24 800, respectively. These findings could provide a reference for medical insurance settlement list quality management in other regions.

Objective:To investigate the spatial distribution patterns of local tumor progression（LTP）after microwave ablation（MWA）for colorectal liver metastases（CRLMs）and identify high-risk progression zones.Methods:A total of 471 CRLM lesions from 246 patients treated with MWA at the Chinese PLA General Hospital between September 2009 and March 2022 were retrospectively analyzed. Three-dimensional visualized MRI image fusion technology was employed to evaluate the spatial relationship between ablation margins（AM）and LTP. The liver was partitioned into nine specific anatomical regions. Machine learning（Boruta algorithm）was used to assess the importance of these regions on LTP risk. Multivariate analysis of LTP was performed at the tumor level and at the patient level using the Cox mixed effects model and the Cox regression model，respectively.Results:LTP occurred in 115 lesions，with an LTP rate of 40.0%（80/200）in ablated lesions which were located in the high-risk progression area，and 12.9%（35/271）in low-risk progression area. Multivariate analysis identified AM < 5 mm，tumor size ≥3 cm and location in high-risk zones as independent risk factors for LTP. Notably，LTP in high-risk zones predominantly clustered around the ablation needle tip.Conclusions:Post-MWA LTP in CRLMs exhibits distinct spatial clustering，particularly at the needle tip within high-risk progression zones. These findings provide critical insights for optimizing ablation strategies and improving clinical outcomes.

OBJECTIVE: To explore the genetic etiology of four fetuses with Uniparental disomy (UPD), and analyze their causes. METHODS: Four fetuses undergoing prenatal diagnosis at Wenzhou Central Hospital between November 2021 and July 2024 were selected as the study subjects. Genetic testing and diagnosis were carried out through G-banded chromosomal karyotyping, single nucleotide polymorphism array (SNP-array) and methylation multiplex ligation-dependent probe amplification (MS-MLPA). This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.: L2024-11-028). RESULTS: The four cases of pathogenic UPD had involved chromosomes 2, 11, 15 and 16, respectively, of which 2 cases were accompanied by fetal ultrasound abnormalities, One fetus was shown a high risk by serological screening, while another showed a high risk by non-invasive DNA testing. The karyotype of fetus 1 was 45,X?,rob(13;15)(q10;q10), and its parents had both carried a Robertsonian translocation involving chromosomes 13 and 15, whilst the karyotypes of other three fetuses were all normal. Pedigree analysis indicated that the UPDs in three cases were paternally derived, and the remaining one was unknown. The causes of the four cases included imprinting syndrome in two cases, autosomal recessive disorder in one case, and cryptic mosaic trisomy in one case. CONCLUSION The clinical phenotypes of UPD are diverse, and the mechanisms are complex. Combined chromosomal karyotyping, SNP-array, MS-MLPA and other technologies are required to make a clear diagnosis for prenatal genetic counseling and postnatal management.
Humans ; Uniparental Disomy/diagnosis* ; Female ; Pregnancy ; Prenatal Diagnosis/methods* ; Polymorphism, Single Nucleotide/genetics* ; Karyotyping ; Adult ; Genetic Testing ; Male ; Fetus

Objective:To investigate the clinical value of puncture biopsy for the diagnosis of vessels that encapsulate tumor clusters (VETC) and macrotrabecular-massive (MTM) subtypes of hepatocellular carcinoma (HCC).Methods:One hundred and eighty-four patients with HCC who underwent surgical resection at the Fifth Medical Centre of Chinses PLA General Hospital from November 2023 to July 2024 were prospectively collected, including 154 males and 30 females, aged (57.1±8.6) years. By simulating the clinical puncture procedure, puncture biopsy tissue specimens were obtained postoperatively from the patient's isolated tumors. The puncture biopsies and surgical resection specimens were stained with HE and CD34, and evaluated for VETC and MTM. Patients were divided into two groups based on the histopathological VETC results of surgically resected specimens: the VETC-positive group ( n=41) and the VETC-negative group ( n=143); and two groups based on the histopathological MTM results of surgically resected specimens: the MTM-positive group ( n=39) and the MTM-negative group ( n=145). Clinical data such as gender, age, tumor length, and alpha-fetoprotein (AFP) were recorded. Logistic regression analysis was performed to screen the risk factors of VETC and MTM. Evaluating the diagnostic efficacy of puncture biopsy for VETC and MTM. Results:The results of multivariable logistic analysis showed that puncture biopsy VETC-positive ( OR=63.97, 95% CI: 16.28-251.29), grade of M2 microvascular invasion ( OR=5.07, 95% CI: 1.31-19.59) and tumor length ≥5 cm ( OR=3.42, 95% CI: 1.11-10.52) were the risk factors for VETC-positive (all P<0.05); whereas the risk factors for MTM-positive were only puncture biopsy MTM-positive ( OR=34.78, 95% CI: 12.06-100.29, P<0.001). Puncture biopsy correctly diagnosed VETC subtype in 163 patients with a diagnostic accuracy of 0.89, sensitivity of 0.61, specificity of 0.97, positive predictive value (PPV) of 0.83, and negative predictive value (NPV) of 0.90; MTM subtype was correctly diagnosed in 164 patients with a diagnostic accuracy of 0.89, sensitivity of 0.72, specificity of 0.94, PPV of 0.76, and NPV of 0.93. Using the three indicators of puncture biopsy diagnosis, tumor length and AFP level as a combined indicator, the accuracy to diagnose VETC was 0.83, sensitivity was 0.71, specificity was 0.87, PPV was 0.60, and NPV was 0.91; and the combined indicator diagnosis of MTM had a diagnostic accuracy of 0.85, a sensitivity of 0.82, specificity of 0.86, PPV of 0.68 and NPV of 0.95. Conclusion:Puncture biopsy has high specificity and accuracy in the diagnosis of VETC and MTM subtypes, but the sensitivity is relatively limited, and the role of puncture combined with clinical factors in improving diagnostic efficacy is limited.

Objective:A nomogram model for predicting double positivity of microvascular invasion (MVI) and vascular endothelial-to-mesenchymal transition (VETC) in patients with hepatocellular carcinoma (HCC) was constructed and its predictive performance was evaluated.Methods:A retrospective analysis was conducted on 326 HCC patients who were treated at the Third People's Hospital of Nantong and the First Affiliated Hospital of Soochow University from January 2013 to June 2023, including 240 males and 86 females, with an average age of (58.7±9.0) years. The 326 patients were randomly divided into a training set ( n=228) and a test set ( n=98) at a ratio of 7: 3 using the random number table method. The training set was divided into a double-positive group ( n=54) and a control group ( n=174) based on whether the HCC patients were double positive for MVI and VETC. Univariate and multivariate logistic regression analyses were performed to identify the influencing factors of double positivity of microvascular invasion in HCC patients, and a nomogram for predicting double positivity of microvascular invasion patterns was constructed based on the multivariate. The predictive performance and clinical net benefit of the nomogram were evaluated using the receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis. Results:There were statistically significant differences in alpha-fetoprotein, gamma-glutamyl transferase, and phosphatidylinositol proteoglycan between the two groups (all P<0.05). Multivariate logistic regression analysis showed that LI-RADS category ( OR=8.58, 95% CI: 1.87-39.38), intratumoral hemorrhage ( OR=2.16, 95% CI: 1.14-4.07), and intratumoral arteries ( OR=2.59, 95% CI: 1.19-5.64) were all influencing factors of double positivity of microvascular invasion patterns in HCC patients (all P<0.05). Based on the multivariate results, a nomogram was constructed. In the training set, the area under the ROC curve for predicting double positivity of microvascular invasion patterns in HCC patients was 0.769 (95% CI: 0.720-0.814). In the test set, the area under the ROC curve for predicting double positivity of microvascular invasion patterns in HCC patients was 0.756 (95% CI: 0.622-0.850). The calibration curve showed a good fit between the predicted model and the ideal curve. Decision curve analysis showed that the clinical applicability was good when the threshold was 0.01-0.80 in the training set and 0.01-0.65 in the test set. Conclusion:The nomogram model based on LI-RADS category, intratumoral hemorrhage, and intratumoral arteries can effectively predict double positivity of microvascular invasion patterns in HCC patients and has good clinical applicability.