Aim To establish a quantitative immunofluorescent bioactivity assay(ICW)for insulin glargine based on CHO-IN-SRB 1284 transgenic cells,and to study its equivalence with in-sulin bioassay of Ch.P.Methods The cells were diluted 25 times with 1.5 × 108 L-1 cell density plates and 1 500 μmol·L-1 insulin glargine,and then diluted with a 3-fold gradient se-ries.The cells were stimulated in microporous plates for 20 min.After fixation,permeation and antibody incubation.Quantitative immunofluorescence biological activity was detected by odyssey two-color infrared fluorescence imaging system.Results There was a good dose-effect relationship between the concentration of insulin glargine in ICW and its relative potency.The method had good specificity,and the relative accuracy,intermediate preci-sion and linearity met the requirements.The relative deviation of biological activity results of 7 batches of insulin glargine samples measured by the two methods was less than 10％.The results were analyzed by SPSS and SAS software,which showed that the methods were correlated and equivalent.Conclusions The quantitative immunofluorescence assay for the biological activity of insulin glargine can be established.The method has good spe-cificity,high accuracy and precision,and has correlation and e-quivalent with biotiter assay,which can be applied to in vitro ef-ficacy evaluation and quality control of insulin glargine.

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]

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Tuberculosis （TB） and human immunodeficiency virus infection / acquired immune deficiency syndrome （HIV/AIDS） are both serious global public health threats. Early detection of infected persons and/or patients through TB/HIV bi-directional screening is crucial for prevention and control strategy in China and globally. In recent years， with the promotion and application of new TB and HIV detection technologies worldwide， TB/HIV bi-directional screening technologies and strategies have made remarkable changes. This expert consensus introduces the significance and challenges of TB/HIV bi-directional screening， summarizes important progress of research and applications， and makes recommendations on screening measures and procedures to further strengthen TB/HIV bi-directional screening in China.

Objective Little is known about the association between whole-blood nicotinamide adenine dinucleotide(NAD+)levels and nabothian cysts.This study aimed to assess the association between NAD+levels and nabothian cysts in healthy Chinese women. Methods Multivariate logistic regression analysis was performed to analyze the association between NAD+levels and nabothian cysts. Results The mean age was 43.0±11.5 years,and the mean level of NAD+was 31.3±5.3 μmol/L.Nabothian cysts occurred in 184(27.7%)participants,with single and multiple cysts in 100(15.0%)and 84(12.6%)participants,respectively.The total nabothian cyst prevalence gradually decreased from 37.4%to 21.6%from Q1 to Q4 of NAD+and the prevalence of single and multiple nabothian cysts also decreased across the NAD+quartiles.As compared with the highest NAD+quartile(≥34.4 μmol/L),the adjusted odds ratios with 95%confidence interval of the NAD+Q1 was 1.89(1.14-3.14)for total nabothian cysts.The risk of total and single nabothian cysts linearly decreased with increasing NAD+levels,while the risk of multiple nabothian cysts decreased more rapidly at NAD+levels of 28.0 to 35.0 μmol/L. Conclusion:Low NAD+levels were associated with an increased risk of total and multiple nabothian cysts.

Objective: To understand the influence of junior middle school students health literacy on knowledge, belief and behavior of COVID-19 in rural areas of Jiangxi Province, and to enhance junior middle school students ability to deal with public health emergencies. Methods: Stratified cluster random sampling was used to investigate the health literacy, knowledge level and behavior of COVID-19 protection of 4 311 grade 7 to grade 8 students in rural areas of Jiangxi Province; Chi square test and Logistic regression analysis were used to analyze the correlation between junior high school students health literacy and COVID-19 protection knowledge, belief and behavior. Results: The rate of health literacy of junior middle school students in rural areas was 18.21 %( n =785), the reported rate of intermediate level was high ( n =2 454, 56.92%), and the reported rate of junior high school students at a low level of health literacy was 24.87%( n =1 072). The rate of junior middle school students in rural areas with good COVID-19 protection knowledge was 63.49%, the rate of positive protection attitude was 74.25%, and the rate of good protection behavior was 85.36%; Rate of COVID-19 protection knowledge ( OR=4.85, 95%CI =3.80-6.18) and positive rate of protection attitude of high level health literacy ( OR=44.07, 95%CI =24.57-79.05), protective behavior possession rate ( OR=25.99, 95%CI = 19.67-34.35) were higher than those with low level of health literacy( P <0.01). Conclusion Health literacy is associated with COVID-19 protection knowledge, belief and behavior in rural junior high school students of Jiangxi Province, the findings provide direction for junior middle school students to improve their ability to deal with public health emergencies.

OBJECTIVE: To explore whether casticin (CAS) suppresses stemness in cancer stem-like cells (CSLCs) obtained from human cervical cancer (CCSLCs) and the underlying mechanism. METHODS: Spheres from HeLa and CaSki cells were used as CCSLCs. DNA methyltransferase 1 (DNMT1) activity and mRNA levels, self-renewal capability (Nanog and Sox2), and cancer stem cell markers (CD133 and CD44), were detected by a colorimetric DNMT activity/inhibition assay kit, quantitative real-time reverse transcription-polymerase chain reaction, sphere and colony formation assays, and immunoblot, respectively. Knockdown and overexpression of DNMT1 by transfection with shRNA and cDNA, respectively, were performed to explore the mechanism for action of CAS (0, 10, 30, and 100 nmol/L). RESULTS: DNMT1 activity was increased in CCSLCs compared with HeLa and CaSki cells (P<0.05). In addition, HeLa-derived CCSLCs transfected with DNMT1 shRNA showed reduced sphere and colony formation abilities, and lower CD133, CD44, Nanog and Sox2 protein expressions (P<0.05). Conversely, overexpression of DNMT1 in HeLa cells exhibited the oppositive effects. Furthermore, CAS significantly reduced DNMT1 activity and transcription levels as well as stemness in HeLa-derived CCSLCs (P<0.05). Interestingly, DNMT1 knockdown enhanced the inhibitory effect of CAS on stemness. As expected, DNMT1 overexpression reversed the inhibitory effect of CAS on stemness in HeLa cells. CONCLUSION CAS effectively inhibits stemness in CCSLCs through suppression of DNMT1 activation, suggesting that CAS acts as a promising preventive and therapeutic candidate in cervical cancer.
Female ; Humans ; Cell Line, Tumor ; HeLa Cells ; Neoplastic Stem Cells/metabolism* ; RNA, Small Interfering/metabolism* ; Uterine Cervical Neoplasms/metabolism*

Objective:To retrospect the blood transfusion status of patients with extensive burns in multiple centers and analyze its influencing factors.Methods:A retrospective case series study was conducted. Clinical data of 455 patients with extensive burns who met the inclusion criteria and were admitted to the burn centers of 3 hospitals from January 2016 to June 2022 were collected, including 202 patients from the First Affiliated Hospital of Nanchang University, 179 patients from the Second Affiliated Hospital of Zhejiang University School of Medicine, and 74 patients from the First Affiliated Hospital of Anhui Medical University. The following data were collected from patients during their hospitalization, including infusion of red blood cells, plasma, and platelets during hospitalization; age, gender, body mass index, combined underlying diseases, cause of injury, time of admission after injury, type of admission, total burn area, full-thickness burn area, combination of inhalation injury, combination of other trauma, and combination of pulmonary edema; the blood lactic acid, serum creatinine, total bilirubin, and albumin values within 24 h of admission; combination of bloodstream, wound, lung, and urinary tract infection, and combination of sepsis; the number of escharectomy or tangential excision and skin grafting surgery (hereinafter referred to as surgery) and total surgical blood loss volume; occurrence of hemoglobin<70 g/L, admission to intensive care unit (ICU), conduction of mechanical ventilation and continuous renal replacement therapy (CRRT), length of hospital stay, and prognosis were recorded. In 602 surgeries of patients within 14 days after injury, data including area of escharectomy or tangential excision and skin graft harvesting, duration of operation, and surgical blood loss volume per surgery, operation site, and use of tourniquet and wound graft were collected. Data were statistically analyzed with Mann-Whitney U test, Kruskal-Wallis H test, and Spearman correlation analysis. Combined with the results of single factor analysis and clinical significance, multiple linear regression analysis was performed to screen the independent influencing factors of red blood cell infusion volume and plasma infusion volume, as well as blood loss volume per surgery. Results:During the whole hospitalization period, 437 (96.0%) patients received blood transfusion therapy, including 435 (95.6%) patients, 410 (90.1%) patients, and 73 (16.0%) patients who received transfusion of plasma, red blood cells, and platelets, respectively. The patients were mainly male, aged 18 to 92 years. There were statistically significant differences in the plasma infusion volume among patients with different combination of underlying disease, combination of inhalation injury, combination of other trauma, combination of pulmonary edema, combination of bloodstream infection, combination of wound infection, combination of lung infection, combination of urinary tract infection, combination of sepsis, occurrence of hemoglobin value <70 g/L, admission to ICU, conduction of mechanical ventilation, and conduction of CRRT (with Z values of -2.06, -4.67, -2.11, -6.13, -9.56, -4.93, -8.08, -4.78, -9.12, -6.55, -9.37, -11.46, and -7.17, respectively, P<0.05). The total burn area, full-thickness burn area, blood lactic acid value within 24 h of admission, serum creatinine value within 24 h of admission, albumin value within 24 h of admission, number of surgeries, and total surgical blood loss volume were correlated with the plasma infusion volume of patients (with r values of 0.39, 0.51, 0.14, 0.28, -0.13, 0.47, and 0.56, respectively, P<0.05).There were statistically significant differences in the red blood cell infusion volume among patients with different gender, combination of inhalation injury, combination of other trauma, combination of pulmonary edema, combination of bloodstream infection, combination of wound infection, combination of lung infection, combination of urinary tract infection, combination of sepsis, occurrence of hemoglobin value <70 g/L, admission to ICU, conduction of mechanical ventilation, and conduction of CRRT (with Z values of -2.00, -4.34, -3.10, -4.22, -8.24, -7.66, -8.62, -4.75, -7.42, -9.36, -6.12, and -8.31, -6.64, respectively, P<0.05). The age, total burn area, full-thickness burn area, blood lactic acid value within 24 h of admission, serum creatinine value within 24 h of admission, total bilirubin value within 24 h of admission, number of surgeries, and total surgical blood loss volume were correlated with the red blood cell infusion volume of patients (with r values of 0.12, 0.22, 0.49, 0.09, 0.18, 0.13, -0.15, 0.69, and 0.77, respectively, P<0.05). Combined underlying diseases, full-thickness burn area, combined pulmonary edema, serum creatinine value within 24 h of admission, combined sepsis, conduction of CRRT, number of surgeries, and total surgical blood loss volume were the independent influencing factors for plasma infusion volume during hospitalization in patients with extensive burns (with standardized regression coefficients of 0.09, 0.16, 0.12, 0.07, 0.11, 0.15, 0.31, and 0.26, respectively, P<0.05). Female, full-thickness burn area, serum creatinine value within 24 h of admission, combined sepsis, occurrence of hemoglobin value <70 g/L, conduction of CRRT, and total surgical blood loss volume were the independent influencing factors for red blood cell infusion volume during hospitalization in patients with extensive burns (with standardized regression coefficients of 0.10, 0.12, 0.10, 0.11, 0.05, 0.19, and 0.54, respectively, P<0.05). There were statistically significant differences in blood loss volume per surgery of patients with different surgical site and wound graft (with Z values of -2.54 and -2.27, respectively, P<0.05). The area of escharectomy or tangential excision and skin graft harvesting and duration of operation were correlated with the blood loss volume per surgery of patients (with r values of 0.40 and 0.21, respectively, P<0.05). The area of escharectomy or tangential excision and skin graft harvesting, duration of operation, and active wound grafts were the independent influencing factors for blood loss volume per surgery of patients with extensive burns (with standardized regression coefficients of 0.41, 0.16, and 0.12, respectively, P<0.05). Conclusions:The major factors influencing blood transfusion status in patients with extensive burns are female, combined underlying diseases, full-thickness burn area, serum creatinine value within 24 h of admission, combined pulmonary edema, occurrence of hemoglobin value <70 g/L, combined sepsis, conduction of CRRT, number of surgery, and total surgical blood loss volume. In addition, the area of escharectomy or tangential excision and skin graft harvesting, duration of operation, and active wound grafts indirectly affect the patient's blood transfusion status by affecting the blood loss volume per surgery.

Objective To understand the characteristics and developmental differences between cerebral organoids in vitro and normal cerebral cortices in vivo. Methods 1. Grouping: cerebral cortices in vivo group and cultured cerebral organoids in vitro group. 2. Sample collection: cortical tissues were collected from Kunming mouse embryos at embryonic day 7.5(E7.5), E9.5, E11.5, E14.5, and postnatal day 3 (P3) or P7. Three specimens were taken from each group. Meanwhile, cerebral organoids were cultured with mouse induced pluripotent stem cells (iPSCs), and samples at different culture time point were collected, and more than 3 samples were collected at each time point. 3. Detection method: the distribution of different types of cells in each group of specimens was analyzed by immunofluorescent staining. Results While relative similarities between in vivo cerebral cortical development and the cerebral organoids in vitro were observed, including the histogenesis, and the morphological differentiation of nerve cells and glial cells, the lamellar architecture of cerebral cortex in mouse brain was not observed in cerebral organoids. Conclusion The development of cerebral organoids in vitro has some similarity with body's cortical development. Therefore, cerebral organoids can be used to a substitution of cortex and diseases' models, but improvement of the existing technologies is necessary.

Objective Using event-related potentials(ERPs),to study the effect and mechanism of negative emotion accumulation hepatic insufficiency on working memory in normal people.Methods Fifty subjects in each of the emotionally stable group and emotionally unstable group were given two load tasks(0-back and 1-back)in the N-back paradigm,the reaction time and correct rate were recorded,and the ERPs components N200 and P300 were detected.The latency and amplitude of P300 were analyzed statistically.Results ①Compared with the emotionally stable group,the emotionally unstable group had a longer reaction time(P<0.05).②Compared with the emotionally stable group,the subjects in the emotionally unstable group had prolonged N200 latency,decreased P300 amplitude significantly(P<0.05),and P300 latency had a tendency to extend(P<0.1).Conclusion Long-term accumulation of negative emotions and liver failure in normal people have the performance of decreased working memory,which may be related to the reduction of attention resource allocation and the impairment of cognitive processing function.