ObjectiveTo genotype the specimens from hand-foot-and-mouth disease (HFMD) cases in Dehong Prefecture, Yunnan Province in 2023 by gene sequencing, and to analyze the genetic characteristics of Coxsackievirus A4 (CVA4), CVA6, CVA10, CVA16 and enterovirus A71 (EV-A71), so as to clarify the serotypes of enteroviruses causing endemic HFMD and the genotype composition of CVA4, CVA6, CVA10, CVA16 and EV-A71 prevalent in HFMD, and to provide baseline genotype data for monitoring the dynamic changes of the five virus genotypes. MethodsStool specimens from HFMD cases in Dehong Prefecture in 2023 that tested positive for EV-A71, CVA16 and other enteroviruses by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) were first subjected to gene sequencing and typing of the VP4/VP2 junction region to determine the viral types. Subsequently, the complete VP1 gene of each virus was amplified in two segments using group-specific VP1 gene primers corresponding to enterovirus groups (i.e., enterovirus group A, B, and C) and sequenced (group A virus: primers 486/488 and 487/489, group B virus: primers 490/492 and 491/493, group C virus: primers 494/496 and 495/497). The representative reference sequences of VP1 genes for CVA4, CVA6, CVA10, CVA16 and EV-A71 were downloaded from the literature, and the VP1 gene phylogenetic tree was constructed using MEGA 6.0 software for genetic characterization. ResultsA total of 256 stool specimens were collected from HFMD cases in Dehong Prefecture in 2023, of which 94 tested positive for nucleic acid by RT-qPCR, yielding a positive rate of 36.72%. Of the 94 positive specimens, 83 were identified as group A viruses, and were classified into 5 serotypes (17 strains of CVA4, 3 of CVA6, 6 of CVA10, 49 of CVA16, and 8 of EV-A71). Nine strains were group B viruses, comprising 2 serotypes: 2 strains of echovirus type 3 (E3) and 7 strains of Coxsackievirus B type 5 (CVB5). The remaining 2 strains were group C viruses, both identified as poliovirus type 3 (PV3). Genetic analyses of the VP1 region showed that all 17 CVA4 strains were identified as subgenotype C2, all 3 CVA6 strains as subgenotype D3a, all 6 CVA10 strains as genotype C. Among the 49 CVA16 strains, 46 were identified as subgenotype B1a and 3 as subgenotype B1b, and all 8 EV-A71 strains were identified as subgenotype C4a. ConclusionIn 2023, group A viruses dominated the pathogenic spectrum of HFMD in Dehong Prefecture. Among them, CVA16 had the highest positivity rate. Genetic characteristic analyses showed that CVA4, CVA6, CVA10 and EV-A71 were each composed of a single genotype (subgenotype), while CVA16 was composed of two subgenotypes (B1a and B1b), with B1a being the predominant one. It is recommended that molecular epidemiological surveillance of HFMD pathogens, particularly group A viruses, be continuously strengthened in Dehong Prefecture.

Autoimmune encephalitis is a type of encephalitis mediated by autoimmune mechanisms, with more than half of the patients requiring intensive care due to consciousness disorder, severe movement disorders, autonomic dysfunction, and status epilepticus. In the early stages, the symptoms of autoimmune encephalitis are varied, including cognitive decline, memory impairments, behavioral changes, movement disorders, and seizures. Without timely and appropriate treatment, it may progress into severe autoimmune encephalitis (SAE) due to challenges in early recognition. Early diagnosis and prompt immunotherapy are crucial for determining the prognosis. The current research evidence and related guidelines, discussing aspects such as symptom management, complication management, and immunotherapy strategies for SAE patients were summarized in this article, with the aim of providing effective references for clinical practice.

Autoimmune encephalitis is a type of encephalitis mediated by autoimmune mechanisms, with more than half of the patients requiring intensive care due to consciousness disorder, severe movement disorders, autonomic dysfunction, and status epilepticus. In the early stages, the symptoms of autoimmune encephalitis are varied, including cognitive decline, memory impairments, behavioral changes, movement disorders, and seizures. Without timely and appropriate treatment, it may progress into severe autoimmune encephalitis (SAE) due to challenges in early recognition. Early diagnosis and prompt immunotherapy are crucial for determining the prognosis. The current research evidence and related guidelines, discussing aspects such as symptom management, complication management, and immunotherapy strategies for SAE patients were summarized in this article, with the aim of providing effective references for clinical practice.

Objective:To explore the impact of home enteral nutrition (HEN) on the treatment strategy of patients with high position intestinal fistula.Methods:The clinical and follow-up data of 36 patients with high position intestinal fistula requiring HEN treated in Beijing Tsinghua Changgung Hospital from Jan 2021 to Sep 2023 was retrospectively analyzed.Results:Among the 36 cases, 2 had indwelling nasogastric tubes, 12 had indwelling nasojejunal nutritional tubes, and 22 had percutaneous jejunostomy. The incidence of HEN-related complications in patients was 13.9%, and there were no serious catheter complications.During HEN, high position intestinal fistula healed in 19 cases (52.8%), returned to the hospital for the next stage of intestinal fistula treatment in 11 cases (30.6%), needed to return to the hospital for nutritional support in 1 case (2.8%), and intestinal fistula aggravated to terminate HEN in 2 cases (5.6%).Conclusion:Under the management of professional team, HEN via nasogastric/jejunal nutritional tube or percutaneous jejunostomy is safe and feasible in patients with high intestinal fistula.

Objective To explore the effect of CD8+T cell subsets on the formation of atheroscle-rosis(AS)in mice.Methods Lasso regression and random forest were employed to identify key infiltrating immune cells in AS blood vessels.Apolipoprotein E gene(ApoE)/recombination acti-vation gene 1(Rag1)double knockout(ApoE/Rag1-DKO)mice were constructed,and then fed with high-fat chow to establish an AS model.The ApoE knockout mice were divided into control group(Con group)and high-fat diet group(HFD group),with 3 mice in each group.And the ApoE/Rag1-DKO mice were divided into DKO+S group[(high-fat diet+phosphate buffer saline(PBS)],DKO+Tem group(high-fat diet with CD8+Tem cell infusion),and DKO+Temra group(high-fat diet with CD8+Temra cell infusion),with 3 mice in each group.Oil red O staining and HE staining were conducted to observe the formation of AS plaques in the vascular walls in the Con,HFD,DKO+S,DKO+Tem,and DKO+Temra groups respectively.Results The results of Lasso regression and random forest showed that CD8+Tem was the most important CD8+T cell subset in the formation of AS.Oil red O staining and HE staining displayed that compared with the Con group,the AS plaque area were significantly larger in the HFD group(P＜0.05);com-pared with the HFD group,there was no significant increase in the AS plaque area in the DKO+S group(P＞0.05);compared with the DKO+S group,the AS plaque area were notably increased in the DKO+Tem group and the DKO+Temra group(P＜0.05).Conclusion CD8+T cell sub-sets are closely associated with the formation of AS in mice,and CD84 Tem and CD84 Temra cells significantly induce the formation.

Background: and Purpose Patients presenting with clinical characteristics that are strongly suggestive of neuromyelitis optica spectrum disorders (NMOSD) have a high risk of developing definite NMOSD in the future. Little is known about the clinical course, treatment, and prognosis of these patients with likely NMOSD at disease onset. Methods: This study prospectively recruited and visited 24 patients with the limited form of NMOSD (LF-NMOSD) at disease onset from November 2012 to June 2021. Their demographics, clinical course, longitudinal aquaporin-4 immunoglobulin G (AQP4-IgG) serology, MRI, therapeutic management, and outcome data were collected and analyzed. Results: The onset age of the cohort was 38.1±12.0 years (mean±standard deviation). The median disease duration was 73.5 months (interquartile range=44.3–117.0 months), and the follow-up period was 54.2±23.8 months. At the end of the last visit, the final diagnosis was categorized into AQP4-IgG-seronegative NMOSD (n=16, 66.7%), AQP4-IgG-seropositive NMOSD (n=7, 29.2%), or multiple sclerosis (n=1, 4.2%). Seven of the 24 patients (29.2%) experienced conversion to AQP4-IgG seropositivity, and the interval from onset to this serological conversion was 37.9±21.9 months. Isolated/mixed area postrema syndrome (APS) was the predominant onset phenotype (37.5%). The patients with isolated/mixed APS onset showed a predilection for conversion to AQP4-IgG seropositivity. All patients experienced a multiphasic disease course, with immunosuppressive therapy reducing the incidence rates of clinical relapse and residual functional disability. Conclusions Definite NMOSD may be preceded by LF-NMOSD, particularly isolated/ mixed APS. Intensive long-term follow-up and attack-prevention immunotherapeutic management is recommended in patients with LF-NMOSD.

Pure drug-assembled nanomedicines (PDANs) are currently under intensive investigation as promising nanoplatforms for cancer therapy. However, poor colloidal stability and less tumor-homing ability remain critical unresolved problems that impede their clinical translation. Herein, we report a core-matched nanoassembly of pyropheophorbide a (PPa) for photodynamic therapy (PDT). Pure PPa molecules are found to self-assemble into nanoparticles (NPs), and an amphiphilic PEG polymer (PPa-PEG

Tumor metastasis is responsible for chemotherapeutic failure and cancer-related death. Moreover, circulating tumor cell (CTC) clusters play a pivotal role in tumor metastasis. Herein, we develop cancer-specific calcium nanoregulators to suppress the generation and circulation of CTC clusters by cancer membrane-coated digoxin (DIG) and doxorubicin (DOX) co-encapsulated PLGA nanoparticles (CPDDs). CPDDs could precisely target the homologous primary tumor cells and CTC clusters in blood and lymphatic circulation. Intriguingly, CPDDs induce the accumulation of intracellular Ca

Objective: To detect the enterovirus VP4 and VP1 genes in 510 stool samples collected from hand, foot and mouth disease (HFMD) cases and analyze the phylogenetic characteristics of the entire VP1 genes of coxsackievirus A6 (CV-A6) strains in six prefectures/cities of Yunnan Province in 2018. Methods: Viral RNA was abstracted from the stool samples. VP4 gene sequences were amplified by RT-PCR and sequenced using the MD91/OL68-1 primer pair to identify viral genotypes. Whole VP1 gene sequences were amplified and sequenced using appropriate primer pairs. The whole VP1 gene sequences of CV-A6 reference strains were downloaded from GenBank. MEGA5.2 software was used to analyze the similarity in nucleotide and amino acid sequences between different strains and phylogenetic tree was constructed for analysis of genetic characteristics and molecular epidemiology. Results: VP4 and VP1 gene sequences were obtained from 57 out of 510 stool samples with a positive rate of 11.17% (57/510). There were 43 CV-A6 (8.43%, 43/510), six CV-A10 (1.17%, 6/510), two enterovirus A71 (EV-A71, 0.39%, 2/510) and two CV-A9 (0.39%, 2/510) strains. The other four strains were CV-A4 (0.19%, 1/510), CV-A5 (0.19%, 1/510), CV-B1 (0.19%, 1/510) and E11 (0.19%, 1/510). The phylogenetic analysis showed that all 43 CV-A6 strains belonged to sub-genotype D3. Conclusions In the 510 HFMD samples, CV-A6 strains were mostly detected with a detection rate of 8.43% and accounted for 75.44% (43/57) of all isolates, followed by CV-A10 (1.17%, 6/510) and EV-A71 (0.39%, 2/510). There was a large HFMD outbreak mainly caused by CV-A6 in Yunnan Province in 2018. The outbreak was caused by CV-A6 of sub-genotype D3, as was the case with pervious outbreaks in China.

Objective To isolate and analyze the genetic characteristics of a new strain of entero-virus EV-C99 from a healthy child in Yunnan Province in 2016. Methods Virus isolation was performed ac-cording to the World Health Organization ( WHO) recommended procedures. Viral RNA was extracted from the supernatant of cell culture. RT-PCR and sequencing analysis of VP1 gene were used for virus identifica-tion. VP1 sequence was edited and stitched by Sequencher 5. 0 software. The edited sequence was BLAST searched in GenBank and the preliminary result indicated that it was an EV-C99 virus. Nucleotide (nt) and amino acid (aa) identities were calculated by MEGA5. 2 software. Serotype of the virus was identified ac-cording to the standard for enterovirus sequence typing. Results The virus could be amplified by 494/496 and 495/497 primer pairs and the edited sequence was about 1 200 bp. Result of the " BLAST" search showed that it was a new strain of enterovirus EV-C99. Comparative analysis with the prototype strain BAN00-10461 (GenBank ID:EF015008) by MEGA5. 2 software showed that the VP1 gene of that virus was 909 bp and the identities between them were 77. 05% in nt sequence and 90. 04% in aa sequence. According to the standard for enterovirus sequence typing,the virus was a new strain of enterovirus EV-C99 belonging to hu-man enterovirus species C (EV-C). Conclusion A new strain of enterovirus EV-C99 was isolated during an investigation of enteroviruses among healthy children in Yunnan Province in 2016. To our knowledge,this is the third report of EV-C99 in China. Phylogenetic analysis reveals that the EV-C99 Yunnan strain,Xinjiang strains and Shandong strains all belong to genotype B,but group into different clusters,indicating that Chinese strains have diverse genetic characteristics.