In the study,calcium carbonate nanocubes(CCNC)were controllably prepared,and a simple and universal modified enzyme-linked immunosorbent assay(ELISA)method was developed for colorimetric and electrochemical dual-mode detection of lung cancer potential biomarker ferritin.The classic sandwich immunoassay mode was adopted.Monoclonal antibodies were modified on microwell of microplate and CCNC firstly.In the presence of ferritin,the CCNC were well captured on the surface of the microplate through the sandwich immunoreaction mode.Calcium ions released by acid hydrolysis formed a colored complex with the following added chromogenic agent,and the color of the sample solution changed from weak yellow to red.Spectrophotometry could be used for the quantitative detection of ferritin.Under the optimal conditions,the absorbance at 512 nm showed a good linear relationship with the logarithm of concentration of ferritin in the range of 1-250 ng/mL,with detection limit of 0.6 ng/mL,which was comparable that of the traditional ELISA method.In addition,gold nanoparticles(AuNPs)modified electrode-based electrochemical technique was also been introduced into the modified ELISA method.Under the optimized conditions,the linear range for ferritin test was 0.05-100 ng/mL,with an extreme low detection limit of 32 pg/mL(S/N>3).The CCNC probe used here exhibited obvious advantages over the enzyme probes in traditional ELISA technique in terms of stability,simplicity and cost,providing a new type of nanoprobe for ELISA technology.

The patient,a male newborn,was admitted to the hospital 2 hours after birth due to prematurity(gestational age 27+5 weeks)and respiratory distress occurring 2 hours postnatally.After admission,the infant developed fever and elevated C-reactive protein levels.On the fourth day after birth,metagenomic next-generation sequencing of cerebrospinal fluid indicated a positive result for Mycoplasma hominis(9 898 reads).On the eighth day,a retest of cerebrospinal fluid metagenomics confirmed Mycoplasma hominis(56 806 reads).The diagnosis of purulent meningitis caused by Mycoplasma hominis was established,and the antibiotic treatment was switched to moxifloxacin[5 mg/(kg·day)]administered intravenously for a total of 4 weeks.After treatment,the patient's cerebrospinal fluid tests returned to normal,and he was discharged as cured on the 76th day after birth.This article focuses on the diagnosis and treatment of neonatal Mycoplasma hominis purulent meningitis,introducing the multidisciplinary diagnosis and treatment of the condition in extremely preterm infants.[Chinese Journal of Contemporary Pediatrics,2024,26(4):432-436]

Objective To investigate the infection status and changing trend of hepatitis C virus(HCV)infection in hospitalized patients in medical institutions,and provide reference for formulating HCV infection prevention and control strategies.Methods HCV infection surveillance results from cross-sectional survey data reported to China Healthcare-associated Infection(HAI)Surveillance System in 2020 were summarized and analyzed,HCV positive was serum anti-HCV positive or HCV RNA positive,survey result was compared with the survey results from 2003.Results In 2020,1 071 368 inpatients in 1 573 hospitals were surveyed,738 535 of whom underwent HCV test,4 014 patients were infected with HCV,with a detection rate of 68.93％and a HCV positive rate of 0.54％.The positive rate of HCV in male and female patients were 0.60％and 0.48％,respectively,with a statistically sig-nificant difference(x2=47.18,P＜0.001).The HCV positive rate in the 50-＜60 age group was the highest(0.76％),followed by the 40-＜50 age group(0.71％).Difference among all age groups was statistically signifi-cant(x2=696.74,P＜0.001).In 2003,91 113 inpatients were surveyed.35 145 of whom underwent HCV test,resulting in a detection rate of 38.57％;775 patients were infected with HCV,with a positive rate of 2.21％.In 2020,HCV positive rates in hospitals of different scales were 0.46％-0.63％,with the highest in hospital with bed numbers ranging 600-899.Patients'HCV positive rates in hospitals of different scales was statistically signifi-cant(X2=35.34,P＜0.001).In 2020,12 provinces/municipalities had over 10 000 patients underwent HCV-rela-ted test,and HCV positive rates ranged 0.19％-0.81％,with the highest rate from Hainan Province.HCV posi-tive rates in different departments were 0.06％-0.82％,with the lowest positive rate in the department of pedia-trics and the highest in the department of internal medicine.In 2003 and 2020,HCV positive rates in the depart-ment of infectious diseases were the highest,being 7.95％and 3.48％,respectively.Followed by departments of orthopedics(7.72％),gastroenterology(3.77％),nephrology(3.57％)and general intensive care unit(ICU,3.10％)in 2003,as well as departments of gastroenterology(1.35％),nephrology(1.18％),endocrinology(0.91％),and general intensive care unit(ICU,0.79％)in 2020.Conclusion Compared with 2003,HCV positive rate decreased significantly in 2020.HCV infected patients were mainly from the department of infectious diseases,followed by departments of gastroenterology,nephrology and general ICU.HCV infection positive rate varies with gender,age,and region.

Objective:Different methods were used to determine the dissolution of piroxicam patch,and the disso-lution results were evaluated,in order to select a determination method that could more accurately reflect the drug release process of piroxicam patch,so as to provide a reference for the scientific and accurate evaluation of drug quality.Methods:The liquid chromatography method for the detection of piroxicam was established,and the 24 hour dissolution curves of piroxicam patch were investigated by paddle over disk method,rotating cylinder method and vertical diffusion cell method,and the dissolution curves were compared by f1 difference factor method,f2 simi-larity factor method and Weibull model fitting,and the in vitro dissolution behavior of different methods was evalua-ted.Results:Piroxicam had a good linear relationship in the range of 1-150 μg·mL-1(r=1.000),the accura-cy was 100.9％(n=9),the precision was 1.7％(n=9),and the sample solution was stable within 72 hours.The results of the comparison of dissolution curves showed that the dissolution of piroxicam patch was more in line with the Weibull model.Under the same conditions of dissolution medium and temperature,there was little differ-ence between the paddle over disk method and rotating cylinder method,and there was a possibility of substitution for each other,and there were significant differences between the vertical diffusion cell method and the other two methods.Conclusion:The vertical diffusion cell method is more in line with the dissolution process of drugs in actual use,and provides more references for quality evaluation.

Objective:To investigate the application value of virtual simulation experiment in the teaching of Laboratory Sciences for Virus Infection Testing.Methods:Undergraduate students(n=58)majoring in Health Inspection and Quarantine of grade 2020 were selected as the research objects.The"Virtual simulation experiment of influenza virus isolation and identification in BSL-2 laboratory"was included in the teaching of Laboratory Sciences for Virus Infection Testing.The application effect of the virtual simulation experiment was evaluated by a self-designed questionnaire.Results:A total of 50 students completed the questionnaire effectively.After completing the virtual simulation experiment,96.0%of the students thought that the simulation degree of the experiment was high.Students could understand the instrument composition,internal structure,operation process,and matters needing attention.And 52.0%of the students thought that the experiment had an impact on autonomous learning.Students believed that the experiment could improve their interest in the course(88.0%),increase their mastery of theoretical knowledge(60.0%),and improve their ability to analyze and solve problems(46.0%).And 72.0%of the students thought that the running speed of the virtual simulation experiment was slow.The top three advantages of the experiment evaluated by students were easy operation,no damage to the instrument and no worry about the dangers in the experiment,and helpful to understand the knowledge.The top three disadvantages of this experiment were no improvement in practical ability,no familiarity with the use of instruments,different feelings from the real experiment and too idealized experimental process.And 66.0%of the students thought that virtual simulation experiments should complement traditional experimental teaching.Conclusions:Virtual simulation experiment can carry out dangerous experiments that are difficult to perform in traditional laboratories.It can improve students'mastery of theoretical knowledge,enhance their interest in learning,and improve their ability to analyze and solve problems.However,there are still some problems in its application,which should be combined with traditional experimental teaching to improve the teaching effect.

Blood stasis syndrome is one of the core clinical syndrome of rheumatoid arthritis (RA), but the biological connotation of this syndrome is not clear, and there is a lack of disease improved animal models that match the characteristics of this disease and syndrome. The aim of this study was to screen the candidate biomarker gene set of blood stasis syndrome of RA, reveal the biological connotation of this syndrome, and explore and evaluate the preparation method of the improved animal model based on the characteristics of "disease-syndrome-symptom". The study was approved by the ethics committee of Guang'anmen Hospital, Chinese Academy of Traditional Chinese Medicine (No. 2019-073-KY-01) and the First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine (No. TYLL2021[K]018), and the study subjects gave their informed consent. Animal welfare and experimental procedures followed the regulations of the Experimental Animal Ethics Committee of the Chinese Academy of Traditional Chinese Medicine (No. IBTCMCACMS21-2207-01). The whole blood samples were collected clinically from RA patients with blood stasis syndrome (3 cases) or other syndromes (7 types, 3 cases/type), and healthy volunteers (4 cases), and then transcriptome sequencing, KEGG, gene set enrichment analysis (GSEA) and weighted correlation network analysis (WGCNA) analysis were performed. 126 pivotal genes were screened, and their functional annotation results were significantly enriched in "immune-inflammation" related pathways and lipid metabolism regulation (sphingolipids, ether lipid metabolism and steroid biosynthesis). Syndrome-symptom mapping of hub gene set to the TCM primary and secondary symptoms, Western phenotypic symptoms and pathological links showed that joint tingling, abnormal joint morphology, petechiae and abnormal blood circulation are representative of blood stasis syndrome of RA. The results of the improved animal model showed that the rats in the collagen-induced arthritis + adrenaline hydrochloride (CIA+Adr) 3 model group had increased blood rheology, coagulation, platelet function and endothelial function abnormalities compared with the CIA-alone model group, suggesting that the rats with blood stasis syndrome of RA may be in a state of "blood stasis". The results of the study can help to advance the objective study of the evidence of blood stasis syndrome in RA, and provide new ideas for the establishment of an animal model that reflects the clinical characteristics of the disease and syndrome.

Objective To understand the distribution and antimicrobial resistance changes of bacteria isolated from pleural and peritoneal effusion in Hunan Province,and provide reference for correct clinical diagnosis and rational antimicrobial use.Methods Data reported by member units of Hunan Provincial Antimicrobial Resistance Survei-llance System from 2012 to 2021 were collected.Bacteria antimicrobial resistance surveillance method was imple-mented according to technical scheme of China Antimicrobial Resistance Surveillance System(CARSS),and WHO-NET 5.6 software was used to analyze the data of bacteria isolated from pleural and peritoneal effusion as well as antimicrobial susceptibility testing results.Results From 2012 to 2021,a total of 28 934 bacterial strains were iso-lated from specimens of pleural and peritoneal effusions from member units of Hunan Provincial Antimicrobial Re-sistance Surveillance System,with 5 752 strains from pleural effusion and 23 182 from peritoneal effusion.The top five bacteria isolated from pleural effusion were Escherichia coli(n=907,15.8％),Staphylococcus aureus(n=535,9.3％),Klebsiella pneumoniae(n=369,6.4％),Staphylococcus epidermidis(n=452,7.9％),and Staphy-lococcus haemolyticus(n=285,5.0％).The detection rate of methicillin-resistant Staphylococcus aureus(MR-SA)from pleural effusion was 24.3％-39.2％,and that of methicillin-resistant coagulase negative Staphylococcus(MRCNS)was 58.8％-77.1％.The top five bacteria isolated from peritoneal effusion were Escherichia coli(n=8 264,35.6％),Klebsiella pneumoniae(n=2 074,9.0％),Enterococcus faecium(n=1 458,6.3％),Staphylo-coccus epidermidis(n=1 383,6.0％),and Pseudomonas aeruginosa(n=1 152,5.0％).The detection rate of MRSA from peritoneal effusion was 22.1％-52.4％,which presented a decreasing trend(P=0.004).The detec-tion rate of MRCNS was 60.4％-79.4％.The resistance rates of Enterobacterales from peritoneal effusion to ce-fazolin,cefuroxime,ceftriaxone and cefepime all showed decreasing trends(all P＜0.05).Vancomycin-,linezo-lid-,and teicoplanin-resistant Staphylococcus strains were not found in pleural and peritoneal effusions.The resis-tance rates of Enterococcus faecium to most tested antimicrobial agents were higher than those of Enterococcus fae-calis.The resistance rates of Enterobacterales to imipenem and meropenem were ≤8.5％.The resistance rates of non-fermentative Gram-negative bacilli to imipenem and meropenem were ≤43.3％.Conclusion The data structure of Hunan Antimicrobial Resistance Surveillance System for pleural and peritoneal effusions from 2012 to 2021 is relatively complete.The constituent and antimicrobial susceptibility of isolated pathogenic bacteria vary in different years.

Objective To observe the value of preoperative CT enterography(CTE)for predicting the necessity of enterectomy in ovarian cancer(OC)patients for achieving optimal debulking surgery(ODS).Methods Preoperative CTE data of 49 patients with pathologically confirmed OC were retrospectively analyzed,including 13 cases received neoadjuvant chemotherapy(NACT)and 36 cases did not.The length(L)and circumference(C)of muscular involvement shown on CTE were taken as criteria for judging the necessity of enterectomy in OC patients([degree 0:no muscular involvement,degree 1:L＜3.0 cm and C＜1/3,degree 2:L≥3.0 cm[A]or C≥1/3[B]).The patients were followed up for at least 1 year after surgery,and the actual operation results were recorded to estimate the ideal surgical strategy.The consistency of CTE evaluated necessity of enterectomy and the ideal surgical strategy was analyzed.Results For patients not undergoing NACT,according to degree 2 A or B showed on CTE,predicted results had excellent agreement with the ideal surgical strategy(Kappa=0.852,P＜0.001),while those according to degree 1 or 2,2 A,2 B were all in good agreement with the ideal surgical strategy(Kappa=0.684-0.771,all P＜0.001).For patients undergoing NACT,the predicted results of degree 2 muscular involvement showed on CTE before NACT had excellent consistencies with the ideal surgical strategy(all Kappa=0.806,all P=0.003).The predicted results of degree 2 muscular involvement showed on CTE after NACT had moderate consistencies with the ideal surgical strategy(all Kappa=0.581,all P=0.021).Conclusion Preoperative CTE could be used to predict the necessity of enterectomy in OC patients for achieving ODS.

Objective:To investigate the clinical effect of using free bilateral anterolateral thigh flaps(ALTF) in series to repair large area soft tissue defects of forearm.Methods:The clinical data of 11 patients with large soft tissue defects of forearm admitted in the Department of Plastic(Repair and Reconstruction) Surgery, Lishui Hospital of Zhejiang University from March 2014 to December 2021 were retrospectively analyzed, including 8 males and 3 females. Aged 36 to 68 years old, with an average of 48 years old. VSD treatment was performed after debridement, and until fresh removed 3 to 5 days after the operation. Until the wound was fresh. The wound was repaired with free bilateral ALTF in series until fresh. The size of the forearm wound was 18 cm×15 cm-28 cm×13 cm. The cut area of a single flap was 10 cm×8 cm-20 cm×13 cm. The series of bilateral flaps: One of the flaps was used as the proximal flap, and its vascular pedicle was anastomosed with the arteries and veins of the recipient area. The other flap was used as the distal flap, and the arteries and veins between the 2 ALTFs on both sides were anastomosed. The vascular pedicle beyond the distal flap was ligated or anastomosed to the distal end of the ulnar artery or the distal end of the radial artery. The flap and the surrounding skin of the recipient area were sutured immediately. The donor sites of the flap was closed directly. Periodic and regular outpatient follow-up was performed after operation and the clinical efficacy was analyzed.Results:All the flaps successfully survived after the surgery. The postoperative follow-up lasted for 6-18 months, 12 months in average. The flaps survived well with good soft texture, without swelling, the capillary reaction time was normal, without surface ulceration, in rosy colour and restored protective sensations. The hand function of the affected limb recovered well. The wound at donor sites healed well without complications. At the last follow-up, the hand function of the affected limb was evaluated by the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, the result was 7 in excellent and 4 in good. The patients were satisfied with the flap and the therapeutic effect.Conclusion:The free bilateral ALTF in series can be used to repair a wound surface with large area, and the donor site can be closed at the same time. It is an effective method to repair large soft tissue defect of forearm.

Objective To explore olanzapine effect on the cognitive function and neuronal damage of aged schizophrenic rats based on the PI3 K/Akt signaling pathway. Methods Ten-week-old SD rats were randomly divided into a blank control group（n=12） and a modeling intervention group（n=48）. The modeling group were injected with didroxapine maleate [MK-801,0.2 mg/（kg·d）] for 14 days. And the model was evaluated by general behavioral studies to determine the success of model building. The model rats were randomly divided into model group and low, medium, and high dose olanzapine groups [10, 20, 40 mg/（kg·d）], each with 12 rats. The control group and model group were given distilled water; the low, medium, and high dose olanzapine groups were given olanzapine for 21 days. The stereotyped lines were scored by the standard of Sams Dodd and Hoffman, the cognitive evaluation of the rats was performed by the Morris water maze, and the levels of interleukin-6（IL-6） and tumor necrosis factor-α（TNF-α） in the serum were determined by ELISA. The activities of dihydrokaempferol（Ach） and acetyl cholinesterase（AchE）in brain tissue were detected by acetylcholinesterase activity assay kit. Rat brain tissue PI3 K, Akt, mammalian target of rapamycin（mTOR） mRNA expression levels were detected by Real-time PCR. Results Compared with the model group, the stereotyped behavior and ataxia scores, escape latency, number of crossing platforms, serum levels of IL-6, TNF-α, AchE, phosphorylated PI3 K（p-PI3 K）, phosphorylated Akt（p-Akt） protein expression decreased（P＜0.05 or P＜0.01）, while brain tissue Ach, PI3 K, mTOR and phosphorylated mTOR（p-mTOR） protein content increased（P＜0.05 or P＜0.01） in the low, medium and high dose olanzapine groups. The content of Akt was increased in the low-dose group. Compared with the model group, Akt and mTOR mRNA in the brain tissue of rats in the low, medium, and high-dose alanzapine groups expression levels were down-regulated（P＜0.05 or P＜0.01）. PI3 K mRNA in the brain tissue of rats in the low, medium, and high-dose alanzapine groups expression levels were down-regulated（P＜0.05 or P＜0.01）. Conclusion Olanzapine can reduce stereotyped behavior and ataxia scores, escape latency, number of crossing platforms, IL-6, TNF-α, AchE and increase Ach content and regulate the PI3 K/Akt signaling pathway to relieve the schizophrenia.