Aim To generate a dual-reporter mouse model for tracing microglia and neurons to analyze their dynamic changes in real-time and to investigate the functional role of microglia-neuron interactions through microglial depletion.Methods Mi-croglia reporter mice were crossed with neuron reporter mice to generate Tmem119-e(2A-tdTomato-2A-DTR);Thy1-GFP dual-reporter mice.Genotyping was performed using tail DNA.Con-focal and two-photon microscopy were used to examine neuronal morphology.In vivo two-photon imaging was employed to observe the distribution of microglia and neurons in the brain.Diphtheria toxin(DT)was intraperitoneally injected to deplete microglia.Results The dual-reporter mice were successfully generated with the correct genotype.Both microglia and neurons were visu-alized,and microglia were specifically depleted by DT.Conclu-sions The successful establishment of a dual-reporter mouse model for tracing neurons and microglia will facilitate real-time in vivo observation of microglial and neuronal changes under physiological or pathological conditions,elucidating their specific roles in brain homeostasis.

Aim To generate a dual-reporter mouse model for tracing microglia and neurons to analyze their dynamic changes in real-time and to investigate the functional role of microglia-neuron interactions through microglial depletion.Methods Mi-croglia reporter mice were crossed with neuron reporter mice to generate Tmem119-e(2A-tdTomato-2A-DTR);Thy1-GFP dual-reporter mice.Genotyping was performed using tail DNA.Con-focal and two-photon microscopy were used to examine neuronal morphology.In vivo two-photon imaging was employed to observe the distribution of microglia and neurons in the brain.Diphtheria toxin(DT)was intraperitoneally injected to deplete microglia.Results The dual-reporter mice were successfully generated with the correct genotype.Both microglia and neurons were visu-alized,and microglia were specifically depleted by DT.Conclu-sions The successful establishment of a dual-reporter mouse model for tracing neurons and microglia will facilitate real-time in vivo observation of microglial and neuronal changes under physiological or pathological conditions,elucidating their specific roles in brain homeostasis.

Objective To investigate the clinical characteristics and prognosis of pneumococcal meningitis(PM),and drug sensitivity of Streptococcus pneumoniae(SP)isolates in Chinese children.Methods A retrospective analysis was conducted on clinical information,laboratory data,and microbiological data of 160 hospitalized children under 15 years old with PM from January 2019 to December 2020 in 33 tertiary hospitals across the country.Results Among the 160 children with PM,there were 103 males and 57 females.The age ranged from 15 days to 15 years,with 109 cases(68.1% )aged 3 months to under 3 years.SP strains were isolated from 95 cases(59.4% )in cerebrospinal fluid cultures and from 57 cases(35.6% )in blood cultures.The positive rates of SP detection by cerebrospinal fluid metagenomic next-generation sequencing and cerebrospinal fluid SP antigen testing were 40% (35/87)and 27% (21/78),respectively.Fifty-five cases(34.4% )had one or more risk factors for purulent meningitis,113 cases(70.6% )had one or more extra-cranial infectious foci,and 18 cases(11.3% )had underlying diseases.The most common clinical symptoms were fever(147 cases,91.9% ),followed by lethargy(98 cases,61.3% )and vomiting(61 cases,38.1% ).Sixty-nine cases(43.1% )experienced intracranial complications during hospitalization,with subdural effusion and/or empyema being the most common complication[43 cases(26.9% )],followed by hydrocephalus in 24 cases(15.0% ),brain abscess in 23 cases(14.4% ),and cerebral hemorrhage in 8 cases(5.0% ).Subdural effusion and/or empyema and hydrocephalus mainly occurred in children under 1 year old,with rates of 91% (39/43)and 83% (20/24),respectively.SP strains exhibited complete sensitivity to vancomycin(100% ,75/75),linezolid(100% ,56/56),and meropenem(100% ,6/6).High sensitivity rates were also observed for levofloxacin(81% ,22/27),moxifloxacin(82% ,14/17),rifampicin(96% ,25/26),and chloramphenicol(91% ,21/23).However,low sensitivity rates were found for penicillin(16% ,11/68)and clindamycin(6% ,1/17),and SP strains were completely resistant to erythromycin(100% ,31/31).The rates of discharge with cure and improvement were 22.5% (36/160)and 66.2% (106/160),respectively,while 18 cases(11.3% )had adverse outcomes.Conclusions Pediatric PM is more common in children aged 3 months to under 3 years.Intracranial complications are more frequently observed in children under 1 year old.Fever is the most common clinical manifestation of PM,and subdural effusion/emphysema and hydrocephalus are the most frequent complications.Non-culture detection methods for cerebrospinal fluid can improve pathogen detection rates.Adverse outcomes can be noted in more than 10% of PM cases.SP strains are high sensitivity to vancomycin,linezolid,meropenem,levofloxacin,moxifloxacin,rifampicin,and chloramphenicol.[Chinese Journal of Contemporary Pediatrics,2024,26(2):131-138]

Objective: To analyze the clinical epidemiological characteristics including composition of pathogens , clinical characteristics, and disease prognosis acute bacterial meningitis (ABM) in Chinese children. Methods: A retrospective analysis was performed on the clinical and laboratory data of 1 610 children <15 years of age with ABM in 33 tertiary hospitals in China from January 2019 to December 2020. Patients were divided into different groups according to age,<28 days group, 28 days to <3 months group, 3 months to <1 year group, 1-<5 years of age group, 5-<15 years of age group; etiology confirmed group and clinically diagnosed group according to etiology diagnosis. Non-numeric variables were analyzed with the Chi-square test or Fisher's exact test, while non-normal distrituction numeric variables were compared with nonparametric test. Results: Among 1 610 children with ABM, 955 were male and 650 were female (5 cases were not provided with gender information), and the age of onset was 1.5 (0.5, 5.5) months. There were 588 cases age from <28 days, 462 cases age from 28 days to <3 months, 302 cases age from 3 months to <1 year of age group, 156 cases in the 1-<5 years of age and 101 cases in the 5-<15 years of age. The detection rates were 38.8% (95/245) and 31.5% (70/222) of Escherichia coli and 27.8% (68/245) and 35.1% (78/222) of Streptococcus agalactiae in infants younger than 28 days of age and 28 days to 3 months of age; the detection rates of Streptococcus pneumonia, Escherichia coli, and Streptococcus agalactiae were 34.3% (61/178), 14.0% (25/178) and 13.5% (24/178) in the 3 months of age to <1 year of age group; the dominant pathogens were Streptococcus pneumoniae and the detection rate were 67.9% (74/109) and 44.4% (16/36) in the 1-<5 years of age and 5-<15 years of age . There were 9.7% (19/195) strains of Escherichia coli producing ultra-broad-spectrum β-lactamases. The positive rates of cerebrospinal fluid (CSF) culture and blood culture were 32.2% (515/1 598) and 25.0% (400/1 598), while 38.2% (126/330)and 25.3% (21/83) in CSF metagenomics next generation sequencing and Streptococcus pneumoniae antigen detection. There were 4.3% (32/790) cases of which CSF white blood cell counts were normal in etiology confirmed group. Among 1 610 children with ABM, main intracranial imaging complications were subdural effusion and (or) empyema in 349 cases (21.7%), hydrocephalus in 233 cases (14.5%), brain abscess in 178 cases (11.1%), and other cerebrovascular diseases, including encephalomalacia, cerebral infarction, and encephalatrophy, in 174 cases (10.8%). Among the 166 cases (10.3%) with unfavorable outcome, 32 cases (2.0%) died among whom 24 cases died before 1 year of age, and 37 cases (2.3%) had recurrence among whom 25 cases had recurrence within 3 weeks. The incidences of subdural effusion and (or) empyema, brain abscess and ependymitis in the etiology confirmed group were significantly higher than those in the clinically diagnosed group (26.2% (207/790) vs. 17.3% (142/820), 13.0% (103/790) vs. 9.1% (75/820), 4.6% (36/790) vs. 2.7% (22/820), χ²=18.71, 6.20, 4.07, all P<0.05), but there was no significant difference in the unfavorable outcomes, mortility, and recurrence between these 2 groups (all P>0.05). Conclusions: The onset age of ABM in children is usually within 1 year of age, especially <3 months. The common pathogens in infants <3 months of age are Escherichia coli and Streptococcus agalactiae, and the dominant pathogen in infant ≥3 months is Streptococcus pneumoniae. Subdural effusion and (or) empyema and hydrocephalus are common complications. ABM should not be excluded even if CSF white blood cell counts is within normal range. Standardized bacteriological examination should be paid more attention to increase the pathogenic detection rate. Non-culture CSF detection methods may facilitate the pathogenic diagnosis.
Adolescent ; Brain Abscess ; Child ; Child, Preschool ; Escherichia coli ; Female ; Humans ; Hydrocephalus ; Infant ; Infant, Newborn ; Male ; Meningitis, Bacterial/epidemiology* ; Retrospective Studies ; Streptococcus agalactiae ; Streptococcus pneumoniae ; Subdural Effusion ; beta-Lactamases

OBJECTIVETo investigate the expression of endothelium tight junction protein Claudin-5 and intercellular adhesion molecule CD99 in solid-pseudopapillary neoplasms (SPN) and neuroendocrine tumors of pancreas (P-NET), and their significance in the differential diagnoses.

METHODSImmunohistochemical staining of Claudin-5 and CD99 was performed in 37 cases SPN and 21 cases of P-NET.

RESULTSMembranous Claudin-5 expression was observed in all cases of SPN but was absent in all cases of P-NET. The difference was significant (P < 0.01). In SPN, 91.9% (34/37) of the cases displayed paranuclear dot-like immunoreactivity for CD99; in contrast, 61.9% (13/21) of the cases of P-NET displayed membranous staining (P < 0.01). There was a positive association between the expression of Claudin-5 and CD99 in SPN (r = 0.421,P = 0.001).

CONCLUSIONSAlthough the macroscopic and microscopic features of SPN are quite characteristic, they may not allow confident differentiation from P-NET in all cases, especially when these characteristics are not classical. If necessary, immunostaining for Claudin-5 and CD99 can help to differentiate between these entities.

12E7 Antigen ; Adolescent ; Adult ; Aged ; Antigens, CD ; metabolism ; Carcinoma, Papillary ; metabolism ; pathology ; Cell Adhesion Molecules ; metabolism ; Child ; Claudin-5 ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Neuroendocrine Tumors ; metabolism ; pathology ; Pancreatic Neoplasms ; metabolism ; pathology ; Retrospective Studies ; Tight Junctions ; metabolism ; Young Adult

OBJECTIVETo study the chemical constituents in Astragalus dahuricus.

METHODThe compounds were isolated and purified by column chromatography on silica gel and HPLC, and their structures were elucidated by their spectroscopic evidences.

RESULTSix compounds were identified as: 7, 2'-dihydroxy-3', 4'-dimethoxyisoflavan (1), 2'-hydroxy-3', 4'-dimethoxyisoflavan-7-O-beta-D-glucopyranoside (2), 8, 2'-dihydroxy-7, 4'-dimethoxyisoflavan (3), 7-hydroxy-4'-methoxyisoflavone (4), 7, 3'-dihydroxy-4'-metho-xyisoflavone (5), 9, 10-dimethoxypterocarpan-3-O-beta-D-glucopyranoside (6).

CONCLUSIONCompounds 1-6 were obtained from this plant for the first time.

Astragalus Plant ; chemistry ; Chromatography, High Pressure Liquid ; Isoflavones ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy

OBJECTIVETo determine the feasibility and the effectiveness of HIV risk behavior intervention characterized by initiator taking the lead combined with peer's participation, as to preventing HIV epidemic through promoting condom use and reducing the number of sexual partners among men who have sex with men (MSM) groups.

METHODSSubjects were recruited via peer referral chain. Twelve key MSM were recruited as initiators in bars or other MSM venues in 3 cities of Hefei, Wuhu and Fuyang. Then, each initiator recruited up to 3 MSM to participate and also each of them continued recruiting others. A total of 218 eligible MSM were recruited, and there were four intervention activities conducted. Firstly, twelve initiators were trained according to intervention manual and then intervention activities were implemented by initiators based on their referral chain. Participants were required to complete self-administrated questionnaire at baseline and the third month after intervention finished. The comparison of the results before and after intervention was conducted two months later to see any improvement in HIV/AIDS knowledge, and condom use.

RESULTSOf 218 participants, 170 (77.9%) were followed up in assessment three months later. The results from paired t-Test and Chi Square Test showed that intervention increased HIV/STDs related knowledge (baseline, 14.71 +/- 2.59; follow-up, 16.95 +/- 1.81; t = -10.647, P < 0.01) and the rate of having female sexual partner during previous 2 months (baseline, 17.6%; follow-up, 11.2%; P < 0.01) were of significant differences. Meanwhile, the intervention increased rate of condom use in the last three times of anal intercourse with homosexual partners, casual homosexual partners and primary homosexual partners (baseline, 55.3%, 43.2%, 49.1%; follow-up, 65.2%, 52.2%, 60.9%; chi(2) = 9.979, P < 0.01; chi(2) = 5.797, P < 0.05; chi(2) = 13.082, P < 0.01; respectively) and decreased rate of non-condom use in the last anal intercourse with homosexual partners, casual homosexual partners and primary homosexual partners (baseline, 41.2%, 35.3%, 45.3%; follow-up, 25.3%, 27.1%, 31.2%; P < 0.01, P < 0.05, P < 0.01; respectively) were all of some improvement. Other relevant indicators of 218 participants with 170 followed were compared, excepting the above similar findings, there were no differences in rate of number of female sexual partner during previous 2 months and rate of condom use in the last three anal intercourse with casual homosexual partners and rate of non-condom use in the last anal intercourse with casual homosexual partners.

CONCLUSIONHIV risk behavior intervention based on MSM peer groups is feasible and might increase the condom use among MSM.

Adult ; Condoms ; Contact Tracing ; Feasibility Studies ; HIV Infections ; epidemiology ; prevention & control ; Homosexuality, Male ; Humans ; Male ; Middle Aged ; Risk-Taking ; Sexual Partners

Objective To investigate the situation of dog rabies and analyze it's relationship with human rabies. Methods In Guizhou, Guangxi and Hunan provinces which suffered from rabies most heavily, one or two cities were selected respectively from regions with high-, middle-, low-incidence rate of human rabies as investigation spots where Dogs' brain specimens were collected and detected with both direct immunofluorescence assay (IFA) and RT-PCR. Results A total of 2887 specimens were collected and 66 of these were positive for IFA and RT-PCR. Therefore, the rate of positive specimens was 2.3 %. However, there was not a close relationship in the incidence rate of dog rabies and human rabies. Conclusion Dog's infection situation of rabies contributed to the severe epidemic of human rabies.