Here,5 important pathogens carried by ticks in Maanshan City,Anhui Province,China were identified.In to-tal,642 ticks were collected from 13 villages around Maanshan City and identified by morphological and mitochondrial COI genes.The 16S rRNA gene of Francisella tularensis,ssrA gene of Bartonella,16S rRNA,ompA and ompB genes of Rickett-sia,16S rRNA and gltA genes of Anaplasma,and groEL and rpoB genes of Coxiella were sequenced.Reference sequences were retrieved from a public database.Phylogenetic trees were constructed with MEG A1 1.0 software.In total,36 Rickettsiae isolates were detected in 640 Haemaphysalis longicornis ticks,which included 20 isolates of Rickettsia heilongjian-gensis,16 of Candidatus Rickettsia jingxinensis,2 of Ana-plasma bovis,and 186 of Coxiella-like endosymbiont.R.hei-longjiangensis HY2 detected in this study and Anhui B8 strain,Ca.R.jingxinensis QL3 and those from Shanxi Prov-ince and Jiangsu Province,A.bovis JX4 and those from Shanxi Province were clustered on the same branch.Overall,17 ticks had combined infections and none of the 5 bacteria were detected in two Amblyomma testudinarium ticks.This is the first report of Ca.R.jingxinensis detected in H.longicornis ticks from Anhui Province.It is recommended that the two types of Rickettsia that cause spotted fever and A.bovis should be reported to local health authorities to initiate appropriate prevention and control measures.

Cell cycle analysis is essential for determining the cell proliferation state,studying cell func-tions,and evaluating drug effects.Flow cytometry is a commonly used method for cell cycle detection,with propidium iodide(PI)being the most widely used fluorescein.Nevertheless,various factors may af-fect the test results.Additionally,comparing distributions of immune cell subpopulations across different cell cycle stages can provide valuable insights into immunological responses and disease conditions.In this research,the B16-F10 cell line was used to study the impact of three factors on PI staining-based cell cycle detection:fixation settings,sample preparation conditions,and software analysis.To fix cells,it is suggested to suspend 3 × 106 cells in 300 μL of pre-cooled PBS,add 700 μL of 100％ethanol drop-wise,fix overnight at 4℃ or-20℃,and collect at a low flow rate(400-600 events/s)to ensure collec-tion of at least 3 000 singlets.Furthermore,dual-labeling with 5-ethynyl-2'-deoxyuridine(EdU)and PI can accurately distinguish cell cycle phases.And various immune cell subpopulations can be analyzed without cell sorting by combining surface marker staining with PI and Ki-67 staining.Here we review fac-tors affecting cell cycle identification using the PI staining method and provide a standard operating proto-col for the experiment.We established the method to combine EdU with PI for cell cycle detection and a-nalysis of immune cell subpopulations,thus expanding the approaches for cell cycle detection.

Hereditary hemochromatosis is a rare autosomal genetic disorder that can cause multi-organ dysfunction in the liver,pancreas,spleen,heart and pituitary gland,with diverse clinical manifestations,make the diagnosis difficult.In recent years,with the deepening of clinical understanding and the development of genetic diagnosis tools,the diagnostic rate of this disease has increased significantly.In this paper,we report a case of hereditary hemochromatosis type 3 involving multiple organs and complicated by severe heart failure,aiming to improve the clinicians'understanding of this disease and reduce the leakage and misdiagnosis.

Objective:To understand the current situation of fear of dementia among the elderly in Changsha and its related factors in order to provide empirical evidence for lowering the level of dementia fear among the elderly and formulating effective strategies.Methods:From July to September 2022, 300 elderly people living in communities in Changsha were surveyed using a random sampling method.Data were collected via a questionnaire for general information, the Fear of Dementia Scale, the Social Support Rating Scale, and the Generalized Anxiety Disorder Scale, and multiple linear regression was used to analyze related factors for fear of dementia in the elderly.Results:For elderly people in Changsha, the fear of dementia score was 46.00(33.25, 59.00), the social support score was 40.00(34.00, 44.75), and the generalized anxiety disorder score was 3.00(2.00, 4.00).According to analysis by Spearman's correlation, the level of fear of dementia among the elderly in Changsha was positively correlated to the level of generalized anxiety( r=0.232, P<0.01)and negatively correlated to the level of social support( r=-0.226, P<0.01).Multiple linear regression analysis revealed that level of education( β=-2.385, t=-3.201, P=0.002), family history of dementia( β=10.801, t=4.151, P<0.001), frequency of physical examination( β=-2.601, t=-3.497, P=0.001), and social support( β=-0.347, t=-3.093, P=0.002)were related factors for fear of dementia in the elderly in Changsha. Conclusions:Fear of dementia among the elderly in Changsha is at a moderate level, and there is no widespread generalized anxiety.Family history of dementia, education level, frequency of physical examination, and level of social support are relevant factors for fear of dementia.To lessen the level of fear of dementia, specific intervention methods should be formulated according to the relevant factors.

Objective To investigate the levels of serum folate and vitamin B12(VB12)and their association with the level of neurodevelopment in preschool children with autism spectrum disorder(ASD).Methods A total of 324 ASD children aged 2-6 years and 318 healthy children aged 2-6 years were recruited.Serum levels of folate and VB12 were measured using chemiluminescent immunoassay.The Social Responsiveness Scale and the Childhood Autism Rating Scale were used to assess the core symptoms of ASD children,and the Gesell Developmental Schedule was employed to evaluate the level of neurodevelopment.Results The levels of serum folate and VB12 in ASD children were significantly lower than those in healthy children(P＜0.05).Serum folate levels in ASD children were positively correlated with gross and fine motor developmental quotients(P＜0.05),and serum VB12 levels were positively correlated with adaptive behavior,fine motor,and language developmental quotients(P＜0.05).In ASD children aged 2 to＜4 years,serum folate levels were positively correlated with developmental quotients in all domains(P＜0.05),and serum VB12 levels were positively correlated with language developmental quotient(P＜0.05).In male ASD children,serum VB12 levels were positively correlated with language and personal-social developmental quotients(P＜0.05).Conclusions Serum folate and VB12 levels in preschool ASD children are lower than those in healthy children and are associated with neurodevelopmental levels,especially in ASD children under 4 years of age.Therefore,maintaining normal serum folate and VB12 levels may be beneficial for the neurodevelopment of ASD children,especially in ASD children under 4 years of age.[Chinese Journal of Contemporary Pediatrics,2024,26(4):371-377]

Objective To explore the potential mechanism of carvacrol on inhibiting the formation of biofilm of hy-pervirulent Klebsiella pneumoniae(hvKP).Methods The possible mechanisms of carvacrol were analyzed based on the detection of its effects on the formation and morphology of biofilms,changes in extracellular polysaccharide and capsule polysaccharide content,as well as changes in the expression levels of biofilm-related genes rmpA2,mag A,mrkA,mrkB,and treC of hvKP.Results The minimum inhibitory concentration of carvacrol on hvKP was 512 μg/mL,with an obvious inhibitory effect on the biofilm formation of hvKP,presenting a concentration-depen-dent effect.Under the scanning electron microscope,it was observed that the biofilm structure was loose and the in-tercellular connections were not dense under the intervention of carvacrol.The Congo Red adsorption test and m-hydroxybiphenyl colorimetric method showed that carvacrol could reduce the content of capsule polysaccharides of hvKP,but didn't affect the total extracellular polysaccharide content.Fluorescence quantitative polymerase chain reaction(PCR)showed that under the effect of carvacrol at sub-inhibitory concentration,the synthesis of capsule polysaccharide,expression levels of sugar transport system and pili adhesion-related genes all decreased by more than 50％.Conclusion Carvacrol has a significant inhibitory effect on the formation of biofilm in hvKP,and its mechanism may be related to the decrease of synthesis of capsule polysaccharide as well as expression of biofilm-re-lated genes,such as sugar transport system and pili adhesion.

BACKGROUND:Pressing massage applied to myofascial trigger points(MTrPs)has shown clear effect in relieving pain.However,further research is needed to investigate the effects of different levels of pressure applied during the massage. OBJECTIVE:To investigate the different strength of pressing on MTrPs in rats with chronic pain. METHODS:Sixty SPF-rated male Sprague-Dawley rats were randomly divided into a blank group of 10 rats not involved in MTrPs modeling and 50 rats involved in modeling.The MTrPs model was established in the left medial thigh muscle of rats by blunt strikes combined with centrifugal exercise and 40 rats that met the evaluation criteria after modeling were randomly divided into model group,light press group,medium press group and heavy press group,with 10 rats in each group.The rats in the blank group and the model group were not intervened,while the rats in the light press group,the medium press group and the heavy press group were intervened with a homemade press stimulator with light force(0.3 kg),medium force(0.5 kg)and heavy force(0.7 kg)to MTrPs.The intervention time was 7.5 minutes per session,with one session every other day,totaling seven sessions.Electromyogram,soft tissue tension and mechanical pain threshold were detected by electrophysiological instruments,soft tissue tension tester,and pressure painmeter,respectively.After the intervention,in the blank group,muscle tissue was taken from the inside of the left thigh,while in the other groups,MTrPs tissue was taken.The pathological morphology was observed by hematoxylin-eosin staining,while enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of cyclooxygenase-2,prostaglandin E2 and bradykinin. RESULTS AND CONCLUSION:Compared with the blank group,the model group had lower mechanical pain thresholds,higher soft tissue tone,higher amplitude frequency of spontaneous electrical activity,significant pathomorphological changes,and increased levels of cyclooxygenase-2,prostaglandin E2 and bradykinin(P<0.05).Compared with the model group,the medium press group and the heavy press group showed increased mechanical pain thresholds,decreased soft tissue tension,decreased spontaneous potential frequency amplitude,and decreased levels of cyclooxygenase-2,prostaglandin E2 and bradykinin(P<0.05),and significant recovery on pathomorphological changes.No significant changes in the above indicators were observed in the light press group(P>0.05).Compared with the medium press group,the heavy press group showed better improvement in the above indicators(P<0.05).To conclude,moderate to heavy pressing is often required to alleviate MTrPs pain.

Objective To assess the risk factors for carbapenem-resistant Acinetobacter baumannii(CRAB)bloodstream infection(BSI)and 28-day short-term mortality in elderly patients,and provide reference for the pre-vention and treatment of CRAB BSI.Methods Clinical data of patients aged ≥60 years and diagnosed with AB BSI in a hospital in Yulin City from January 2013 to December 2022 were retrospectively analyzed,including demogra-phic and microbiological characteristics,as well as clinical outcomes of the patients.Variables which were significant in univariate analysis were selected for multivariate analysis using binary logistic regression model and Cox propor-tional hazards model.Independent risk factors for infection were further determined,and survival analysis was per-formed using Kaplan-Meier curve.Results A total of 150 patients were included in the study,out of which 16 pa-tients(10.7％)had CRAB BSI and 134 had carbapenem-sensitive AB(CSAB)BSI.The 28-day short-term mortali-ty of AB BSI in elderly patients was 15.3％(23/150,95％CI:9.6％-21.1％),and the short-term mortality of CRAB BSI was higher than that of CSAB([56.3％,9/16]vs[10.4％,14/134]).Deep venous catheterization(OR:15.598,95％CI:1.831-132.910)and combined infections of other sites(OR:15.449,95％CI:1.497-159.489)were related to CRAB BSI in elderly patients.The independent risk factors for 28-day mortality in elderly patients with AB BSI were hemodialysis(OR:11.856,95％CI:2.924-48.076),intensive care unit admission(OR:9.387,95％CI:1.941-45.385),and pulmonary infection being suspected source of bacteremia(OR:7.019,95％CI:1.345-36.635).Conclusion The occurrence of CRAB BSI in elderly patients is related to the combined infection of other sites and deep vein catheterization.Hemodialysis,admission to ICU,and pulmonary infection being suspected source of bacteremia are independent risk factors for the prognosis of AB BSI in elderly patients.

Objective To investigate the inhibitory effect of mannose on proliferation and apoptosis of HeLa cells of cervical cancer and its effect on Wnt/β-catenin pathway.Methods Cervical cancer HeLa cells were cultured in vitro and divided into control group(without any treatment)and low,medium and high dose experimental groups(cells treated with 5,10 and 25 mmol·L-1 mannose).Cell counting kit 8(CCK-8)and flow cytometry were used to detect cell proliferation and apoptosis.The B cell lymphoma-2(Bcl-2),p21,Bcl-2 associated X protein(Bax)and Wnt/β-catenin pathway-related proteins were detected by Western blot.Results The cell inhibition rates of control group and low,medium,high dose experimental groups were(0.00±0.13)％,(11.25±3.42)％,(23.78±3.41)％and(35.98±4.52)％;apoptosis rates were(4.59±0.35)％,(11.45±1.32)％,(18.47±2.01)％and(25.69±2.43)％;p21 protein expression were 0.21±0.03,0.34±0.04,0.51±0.06 and 0.69±0.05;the Bcl-2 protein expression were 0.89±0.06,0.67±0.04,0.52±0.05 and 0.35±0.06;Bax protein expression were 0.34±0.05,0.49±0.06,0.65±0.07 and 0.81±0.06;Wnt1 protein expression were 0.74±0.07,0.60±0.05,0.46±0.05 and 0.32±0.04;β-catenin protein expression were 0.80±0.06,0.65±0.05,0.47±0.06 and 0.29±0.05.The above indexes showed statistically significant differences between control group and low,medium and high dose experimental groups(all P＜0.05).Conclusion Mannose inhibits proliferation and induces apoptosis of HeLa cells of cervical cancer,and the mechanism may be related to the inhibition of Wnt/β-catenin pathway.

Isocitrate dehydrogenase 1 (IDH1) R132H is the most common mutated gene in grade II-III gliomas and oligodendrogliomas. Instead of activating telomerase (a reverse transcriptase which using RNA as a template to extend telomere length), the majority of IDH1^R132H mutant glioma maintain telomere length through an alternative mechanism that relies on homologous recombination (HR), which is known as alterative lengthening of telomere (ALT).The phenotype of ALT mechanism include: ALT associated promyelocytic leukemia protein (PML) bodies (APBs); extrachromosomal telomeric DNA repeats such as C- and T-loops; telomeric sister chromatid exchange (T-SCE), etc. The mechanism of ALT activation is not fully understood. Recent studies have shown that mutation IDH1 contributes to ALT phenotype in glioma cells in at least three key ways. Firstly, the IDH1^R132H mutation mediates RAP1 down-regulation leading to telomere dysfunction, thus ensuring persistent endogenous telomeric DNA damage, which is important for ALT activation. Spontaneous DNA damage at telomeres may provide a substrate for mutation break-induced replication (BIR)‑mediated ALT telomere lengthening, and it has been demonstrated that RAP1 inhibits telomeric repeat-containing RNA, transcribed from telomeric DNA repeat sequences (TERRA) transcription to down-regulate ALT telomere DNA replication stress and telomeric DNA damage, thereby inhibiting ALT telomere synthesis. Similarly, in ALT cells, knockdown of telomere-specific RNaseH1 nuclease triggers TERRA accumulation, which leads to increased replication pressure. Overexpression of RNaseH1, on the other hand, attenuates the recombination capacity of ALT telomeres, leading to telomere depletion, suggesting that RAP1 can regulate the level of replication pressure and thus ALT activity by controlling TERRA expression. Secondly, the IDH1^R132H also alters the preference of the telomere damage repair pathway by down-regulating XRCC1, which inhibits the alternative non-homologous end joining (A-NHEJ) pathway at telomeres and alters cellular preference for the HR pathway to promote ALT. Finally, the IDH1^R132H has a decreased affinity for isocitric acid and NADP+ and an increased affinity for α ketoglutarate (α‑KG) and NADPH, so that the mutant IDH1^R132H catalyzes the hydrogenation of α‑KG to produce 2-hydroxyglutarate (2-HG)in a NADPH-dependent manner. Because 2-HG is structurally similar to α‑KG, which maintains the trimethylation level of H3k9me3 by competitively inhibiting the activity of the α‑KG-dependent histone demethylase KDM4B, and recruits heterochromatin protein HP1α to heterochromatinize telomeres, and promote ALT phenotypes in cooperation with the inactivating of ATRX. In addition, it has been shown that APBs contain telomeric chromatin, which is essentially heterochromatin, and HP1α is directly involved in the formation of APBs. Based on these studies, this article reviews the mechanism of IDH1^R132H mediated telomere dysfunction and the preference of DNA repair pathway at telomeres in cooperate with ATRX loss to promote ALT, which may provide references for clinical targeted therapy of IDH1^R132H mutant glioma.