Aim To construct CX3CR1GFP transgenic mice based on the Cre/Loxp system,and to analyze the expression efficiency of CX3CR1GFP.Methods Targeted vectors were designed using restriction enzyme-based cloning technology to create a linearized targeted vector for transfecting embryonic stem cells(ES).The ES cells with a deletion of the neomycin resistance gene(neo)were then cloned into blastocysts to generate chimeric CX3CR1+/GFPmice.These mice were subsequently bred with wild-type mice(WT),and repeated backcrossing was performed to obtain CX3CR1GFP/GFP mice.DNA and mRNA from WT and CX3CR1GFP mice were extracted and genotyped using agarose gel electrophoresis.The expression level of CX3CR1 in various tis-sues of the mice was detected by RT-qPCR.Western blot analy-sis was used to analyze the expression of GFP protein in periph-eral blood mononuclear cells(PBMC)and various tissues.The labeling efficiency of immune cells in bone marrow was detected by flow cytometry.The expression of GFP in different mouse tis-sues was observed by immunofluorescence.Results The results of agarose gel electrophoresis showed that the transgenic mouse genotype was CX3CR1GFP/GFP homozygote.RT-qPCR and West-ern blot showed that EGFP were targeted to replace CX3CR1 gene,so CX3CR1 expression was very low in CX3CR1GFP mice,while GFP expression was significantly upregulated.Flow cytom-etry and immunofluorescence showed that GFP effectively marked CX3CR1GFP mice,expressed in various tissues and cells with different expression levels.Conclusion This study con-structs and identifies the CX3CR1GFP genetic reporter mice,and GFP is stably expressed in mice,which provides a foundation for further research into the potential mechanisms of CX3CR1 in im-mune regulation.

AIM:To explore the mechanism of ac-tion of the microtubular inhibitor of CA-4 deriva-tive LGD5 on human cervical cancer HeLa cells.METHODS:HeLa cells were selected and divided in-to blank group,CA-4 positive control group,and dif-ferent concentrations of LGD5 were formed into the experimental group.MTT was used to investi-gate the growth inhibition of LGD5 on HeLa cells and to determine the assay concentration.Cell morphology and apoptosis were observed before and after drug administration by inverted micro-scope and acridine orange staining.Immunofluo-rescence staining was used to examine the effect of LGD5 on microtubules using DAPI.The effect of LGD5 on cell cycle by PI flow cytometry.Protein im-munoblotting was used to examine the effect of LGD5 on cyclins and apoptosis-related proteins.RE-SULTS:MTT experiments showed that LGD5 inhibit-ed HeLa cells in a time-and dose-dependent man-ner.Timed photography and acridine orange stain-ing observed that LGD5 induced apoptosis in HeLa cells and produced significant chromatin agglutina-tion and apoptotic bodies.Inhibition of microtu-bule polymerization in HeLa cells by LGD5 was ob-served by DAPI staining.The PI flow cytometry re-sults showed that LGD5 induced G2/M cycle arrest in HeLa cells,was time-dependent and dose-depen-dent within 12 h,and had a significant difference(P＜0.01),apoptosis was induced after 24 h and it was time-dependent.The results of Western blot show that,LGD5 downregulates Cdc 2 and Cdc25C,up-regulation of p-Cdc 2,and Cyclin B1 and p-histone H3,further verified that LGD5 induced G2/M cycle arrest in HeLa cells,besides,LGD5 caused in-creased Caspase 3 expression in HeLa cells,upregu-lated Caspase 9 and Bax,down-regulation of Pro-caspase 9 and Bcl-2,this result indicates that HeLa cell apoptosis induced by LGD5 is related to the mi-tochondrial pathway.CONCLUSION:The CA-4 deriv-ative LGD5 inhibited microtubule polymerization in HeLa cells,induced their G2/M cycle arrest,and subsequently induced cell apoptosis through the mitochondrial pathway.

Objective:To explore a preliminary method for comprehensive motor function assessment in older adults in order to promote functional maintenance and independent living.Methods:Cross-sectional study.A total of 1, 680 elderly people aged 60 years and above, aged(69.3±6.8)years, were recruited by volunteer recruitment at the Beijing Hospital Physical Examination Centre and Dongcheng District community, and exercise data were collected on 5 sit-ups, 6 m daily stride, 3 m rise and walk, grip strength, back grip test, chair sit forward bending, and three-posture standing, and the method of exercise function assessment was determined by applying statistical analysis combined with expert guidance.Results:Handgrip strength, Five-time-repeated sit-to-stand, usual gait speed, and 3-meter-timed up and go test were selected for motor function assessment(factor loadings: 0.428, 0.767, 0.725, 0.841, respectively).The motor score was calculated by weighted sum of each indicator with the same weight.Through calculating, the motor score significantly decreased with ageing(60-64 years: 13.9, 65-69 years: 13.4, 70-74 years: 12.7, 75-79 years: 11.3, 80+ years: 9.1; F=89.469, P<0.001).Older adults with moderate activities had higher scores than those without moderate activities(13.6 and 12.4, respectively; t=-8.003, P<0.001). Conclusions:A method for motor function assessment in older adults was established, which can be used in the early recognition of motor function impairment, such as sarcopenia, fall, and disability.

Objective:To explore the effect of Zhisanzhen acupuncture on cognition ability and apoptosis in the hippocampus using a murine model of cerebral ischemia.Methods:Thirty-two male C57 mice were randomly divided into a control group, a model group, a sham acupuncture group and a Zhisanzhen group, each of 8. All except the control group received middle cerebral artery occlusion and reperfusion (MCAO/R) surgery. Successful modeling was confirmed using Bederson scoring and 2, 3, 5-triphenyltetrazolium chloride staining. Beginning 24 hours after successful modeling, the Zhisanzhen group was given 30 minutes of acupuncture at the Shenting and bilateral Benshen points with the injection 3mm deep daily for five days The sham acupuncture group was given acupuncture at 5 mm next to the Shenting and bilateral Benshen points 3mm deep. Twenty-four hours after the last session of acupuncture, the rats′ cognition was tested using a water maze. After the experiment the hippocampus was sampled on the affected side and any apoptosis of hippocampal neurons was detected using the terminal deoxynucleotidyl transferase nick-end labeling (TUNEL) assays. Changes in caspase-3, cleaved caspase-3 protein and p38 mitogen-activated protein kinase (MAPK) protein in the hippocampal tissue were evaluated using western blotting. The total serine phosphorylation level of caspase-3 protein was detected using immunoprecipitation.Results:Compared to the model and the sham groups, the average escape latency in the Zhisanzhen group was significantly shorter and the times crossing the platform was significantly greater. There were significantly fewer TUNEL-positive cells in the Zhisanzhen group on average, and less caspase-3 and cleaved caspase-3, but significantly more p38-MAPK and greater serine phosphorylation of its substrate caspase-3.Conclusion:Zhisanzhen acupuncture can ameliorate recognition difficulties after cerebral ischemia. It promotes phosphorylation of caspase-3, limiting its cleavage and activation, and decreasing neuron apoptosis in the hippocampus.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.

Objective To investigate the incidence and risk factors for acute kidney injury(AKI)in children with primary nephrotic syndrome(PNS),as well as the role of neutrophil gelatinase-associated lipocalin(NGAL)and kidney injury molecule-1(KIM-1)in the early identification of AKI in these children.Methods A prospective collection of clinical data from children hospitalized with PNS at the Children's Hospital of the Capital Institute of Pediatrics from January 2021 to October 2022 was conducted.The children were divided into two groups based on the presence of AKI:the AKI group(47 cases)and the non-AKI group(169 cases).The risk factors for AKI in children with PNS were identified by multivariate logistic regression analysis.Urinary KIM-1 and NGAL levels were compared between the AKI and non-AKI groups,as well as among the different stages of AKI.Results The incidence of AKI in children with PNS was 21.8%.Multivariate logistic regression analysis revealed that steroid-resistant nephrotic syndrome,gastrointestinal infections,and heavy proteinuria were independent risk factors for AKI in these children with PNS(P<0.05).Urinary KIM-1 and NGAL levels were higher in the AKI group compared to the non-AKI group(P<0.05),and the urinary NGAL and KIM-1 levels in the AKI stage 2 and stage 3 subgroups were higher than those in the AKI stage 1 subgroup(P<0.017).Conclusions KIM-1 and NGAL can serve as biomarkers for the early diagnosis of AKI in children with PNS.Identifying high-risk populations for AKI in children with PNS and strengthening the monitoring of related risk factors is of significant importance.

Objective To understand the current situation of protective mask wearing behavior of health care workers(HCWs),analyze the influencing factors for the failure to wear medical protective masks in a standard manner,and provide basis for the improvement of mask-wearing related training.Methods From June 2022 to March 2023,staff in a tertiary first-class hospital were selected as the research object.Real-time quantitative fitness testing using aerosol condensation particle counting method was applied to test 5 commonly used medical protective masks available in the market.Fitness factor changes of the testing instrument and assistance from professional per-sonnel were needed to comprehensively estimate the wearing condition of medical protective masks.Participants were surveyed through a self-made general information questionnaire.Heads and faces of participants were scanned by three-dimensional(3D)laser scanning technology,and scanned images were imported into Geomagic Studio 2013 software to measure head and face dimensions.Results A total of 222 HCWs were investigated,991 real-time tests and 208 times of 3D scanning were conducted.221(22.30％)tests showed failure of participants in wearing masks in a standard manner.The non-standard wearing rates of 5 types of medical protective masks were 30.56％,25.62％,25.87％,23.15％,and 7.35％,respectively.The non-standard mask-wearing rates showed statistically significant difference between groups categorized in terms of medical protective masks with different shapes,partici-pants'occupation,time of last training for wearing medical protective masks,and participants'experience in pre-vention and control of respiratory infectious disease(all P＜0.05).There were no statistically significant differences in non-standard mask-wearing rate between groups with different brands and sizes of medical protective masks,as well as gender and department of participants,etc.(all P＞0.05).The body mass index(BMI)was significantly different among participants who wear foldable medical protective masks in the standard and non-standard manner(both P＜0.05).Conclusion Wearing medical protective masks by HCWs in a non-standard manner is influenced by multiple factors.It is recommended to conduct real-time testing before formal quantitative fitness testing,so as to save time and improve testing efficiency.When conducting training on wearing medical protective masks in the fu-ture,targeted training should be provided based on mask shape and focus on logistics personnel,interns,individuals with high BMI,those who have never received training on wearing medical protective masks,and those who have never participated in the prevention and treatment of respiratory infectious diseases.

Objective To observe the efficacy of gray-scale ultrasound-based radiomics for differentiating peripheral pulmonary adenocarcinoma and squamous cell carcinoma.Methods Data of 88 patients with single peripheral lung adenocarcinoma and 58 patients with single peripheral lung squamous cell carcinoma proved pathologically with puncture biopsy and clearly visualized with lung ultrasound were retrospectively analyzed.The patients were divided into training set(n=103)and test set(n=43)at the ratio of 7:3.Based on gray-scale ultrasound of training set,radiomics features associated with differential diagnosis of peripheral lung adenocarcinoma and lung squamous cell carcinoma were extracted and screened.Using 4 different classifiers,including support vector machine(SVM),linear discriminant analysis(LDA),logistic regression(LR)and the least absolute shrinkage and selection operator combined with logistic regression(LASSO-LR),4 corresponding radiomics models were obtained,and the relative best models were selected according to their performances under 10-fold cross validation.The receiver operating characteristic curves were drawn,the areas under the curve(AUC)were calculated to evaluate the differentiating efficacy of each model,and DeLong test was used for the comparison.The differentiating accuracy of models were obtained under the best cutoff value with the maximum Youden index.Results The AUC of SVM,LDA,LR and LASSO-LR radiomics models for differentiating peripheral lung adenocarcinoma and lung squamous carcinoma in test set was 0.864,0.867,0.880 and 0.844,respectively,and no significant difference was found among 4 models(all P＞0.05).Under the best cutoff value of each model,the corresponding accuracy of SVM,LDA,LR and LASSO-LR radiomics models for differentiating peripheral lung adenocarcinoma and lung squamous cell carcinoma was 86.05％,83.72％,88.37％and 86.05％,respectively.Conclusion Radiomics models based on gray-scale ultrasound could be used to differentiate peripheral lung adenocarcinoma and lung squamous cell carcinoma.

Objective To explore the impact of pre positioned nasopharyngeal airway combined with high head pre inhalation of oxygen on lung oxygenation and blood gas analysis indicators in morbid obese patients undergoing laparoscopic weight loss surgery.Methods 100 morbid obesity patients from January 2020 to April 2022 planned to undergo elective laparoscopic weight loss surgery were selected as the study subjects.All the patients were divided into two groups according to the random number table method:group A with a head height of 25° and a pre installed nasopharyngeal airway;group B with a head height of 25° and no pre installed nasopharyngeal airway,with 50 patients in each group.Two groups were pre oxygenated for 3 min before undergoing intravenous anesthesia to induce tracheal intubation.Observe and record the pH value,partial pressure of oxygen in arterial blood(PaO2),partial pressure of carbon dioxide(PCO2),partial pressure of oxygen in arterial blood/fractional concentration of inspiratory oxygen(PaO2/FiO2)as well as the ratio of arterial pressure to alveolar oxygen partial pressure(a/APO2)of the two groups of patients who breathed air(T0)after entering the room,ventilated with mask positive pressure for 3 min(T1),and intubated with trachea for 3 min(T2).Record plateau pressure(Pplat),peak airway pressure(Ppeak),and dynamic lung compliance(Cdyn)at T1,T2,and 5 min after pneumoperitoneum(T3).Record the time for percutaneous arterial oxygen saturation(SpO2)to decrease to 92.0%under different artificial ventilation after tracheal intubation,the time for SpO2 to recover to 96.0%after resumption of ventilation,and the occurrence of adverse reactions.Results Compared with Group B,at time point T1,Group A showed a decrease in PCO2 and an increase in PaO2,with statistically significant differences(all P<0.05);Compared with T0,at time points T1 and T2,PaO2/FiO2 and PCO2 in the two groups were increased,while a/APO2 decreased(all P<0.05).At T1 time point,Pplat and Ppeak in Group A were lower than those in Group B,while Cdyn was higher than that in Group B,with statistical significance(P<0.05);Compared with T1 time point,at T2 and T3 time point,Pplat and Ppeak in Group A increased,while Cdyn decreased,with statistically significant differences(all P<0.05);Compared with T1,Ppeak increased in B groups at T2 time point(P<0.05),while Pplat and Ppeak increased in T3 time point,and Cdyn decreased in B group,with statistical significance(all P<0.05).Compared with Group B,Group A had a longer time for SpO2 to decrease to 92.0%and a shorter time for SpO2 to recover to 96.0%(P<0.01).Conclusion The combination of pre positioned nasopharyngeal airway and high head pre inhalation of oxygen can effectively improve acute respiratory obstruction during induction of general anesthesia insertion in morbid obesity patients,and extend the duration of no ventilation.

Malignant pleural mesothelioma(MPM)is a rare,highly aggressive,and lethal tumor with poor prognosis.Its survival period ranges from four months to one year,and the 5-year survival rate is only about 10%.MPM is highly resistant to chemotherapy,and conventional treatments such as cisplatin combined with pemetrexed or raltitrexed only have a certain effect in about 20%of patients.In recent years,with the continuous in-depth understanding of the genetic variation characteristics of MPM,some progress has been made in the molecular mechanism underlying the chemotherapy resistance of MPM.This article will summarize the research progress of the molecular mechanism underlying the chemotherapy resistance of MPM,including BAP1 gene mutation,microRNA,MTA1-mediated DNA damage repair pathway,GITR-GITRL pathway,TGFa pathway,tumor stem cell,EGFR,and PTEN.The aim of this work is to provide a reference for exploring new therapeutic targets and combined treatment options for MPM.