Objective:To analyze the drug-resistant gene loci of Mycoplasma pneumoniae (MP) using metagenomic next-generation sequencing (mNGS). Methods:From November 2022 to October 2023, 697 clinical samples (including sputum, alveolar lavage fluid and blood) of 686 children with Mycoplasma pneumoniae positive detected by mNGS were retrospectively analyzed. Samples were divided into intensive care unit (ICU) group and non-ICU group, Chi-square test was used to compare groups, and Mann-Kendall trend test was used to analyze the change trend of the detection rate of drug resistance gene loci over time. Results:Of the 697 samples, 164 were from the ICU group and 533 were from the non-ICU group. The detection rate of Mycoplasma pneumoniae resistance gene was 44.3% (309/697), and all detected drug-resistant gene loci of MP were A2063G. The detection rate of Mycoplasma pneumoniae in ICU group was 50.0% (82/164), and the detection rates of Mycoplasma pneumoniae resistance gene loci in sputum, alveolus lavage fluid and blood samples were 75.0% (18/24) and 48.4% (62/128), respectively. The detection rate in sputum was higher than alveolus lavage fluid samples ( χ2=5.72, P=0.017). The detection rate of Mycoplasma pneumoniae in non-ICU group was 42.6% (227/533), the detection rate of Mycoplasma pneumoniae resistance gene loci in sputum and alveolar lavage fluid was 40.0% (16/40), 44.3% (201/454), and no detection rate in blood samples (0/12). There was no significant difference in the detection rate of alveolar lavage fluid and sputum ( χ2=0.27, P=0.602). From November 2022 to October 2023, the detection rate of submitted samples showed an increasing trend month by month (overall: Z=3.99, ICU inspection group: Z=2.93, non-ICU group: Z=3.01, all P<0.01). Among the bacteria commonly detected with Mycoplasma pneumoniae, Streptococcus pneumoniae accounted for the highest proportion, the detection rate was 15.5% (108/697), and Epstein-Barr virus accounted for the highest proportion of 17.6% (123/697). Conclusions:From November 2022 to October 2023, the detection rate of Mycoplasma pneumoniae drug resistance gene loci showed an increasing trend. The detection rate of drug resistance gene loci in sputum samples of ICU group was higher than alveolus lavage fluid. No new drug resistance site were detected.

Objective To investigate the feasibility of granzyme B(GB)promoter-controlled ferritin heavy chain(FTH1)reporter gene expression for monitoring the activation status of chimeric antigen receptor T cells(CAR-T)by magnetic resonance imaging(MRI).Methods Cytotoxic T lymphocytes(CTLs)were screened by Ficoll density gradient centrifugation and flow sorting.The GB promoter and FTH1 gene were ligated together with disialoganglioside 2(GD2)CAR,and lentiviral vectors were transfected into CTLs to construct GD2-CAR-T/pGB-FTH1 cells.GD2-CAR-T/pCMV-FTH1,GD2-CAR-T,and T cells served as control cells.CytoTox96@non-radioactive cytotoxicity was used to detect the killing effect of each group of cells after co-culture with human neuroblastoma cells(SK-N-SH).ELISA was employed to detect the coincubation factor as well as the amount of GB secretion.Western blotting,Prussian blue staining and cellular MRI were applied to detect the expression of the FTH1 gene after co-culture.Results CTLs were successfully obtained,and then GD2-CAR-T/pGB-FTH1,GD2-CAR-T/pCMV-FTH1 and GD2-CAR-T cells were constructed.The killing effect,co-incubation factor and GB secretion of the above 3 groups of cells were significantly higher than those of the T cells,and the level of GB expression was highest at day 1,and then decreased in order at day 3 and day 7 after co-culturing with SK-N-SH cells.The relative expression of FTH1 and iron content of the GD2-CAR-T/pGB-FTH1 cells showed the same trend as GB expression,and the MRI signals were gradually increased.There were no significant differences in the relative expression of FTH1,iron content and MRI signals in the GD2-CAR-T/pCMV-FTH1 cells at all time points.No FTH1 expression or iron aggregation was observed in the GD2-CAR-T and T cells groups.Conclusion MRI based on the FTH1 reporter gene driven by the granzyme B promoter can reflect the GB expression level and tumor killing effect of CAR-T cells,which provides a potential real-time visual means to monitor the cell activation status for CAR-T therapy.

Objective To investigate the current awareness of pharmaceutical service fees among pharmacists in hospitals of Shaanxi province to provide a theoretical basis and decision-making framework for establishing such fees in hospitals of various provinces and cities in the future.Methods A questionnaire survey was conducted among 47 representative hospitals and 53 pharmacists within these hospitals in Shaanxi province.The results were analyzed using differential analysis.Results In most hospitals of Shaanxi province,pharmaceutical services are not provided or not charged for,indicating a lack of practical experience in the establishment of pharmaceutical service fees.Among hospitals that provide and charge for pharmaceutical services,there remains a need for uniformity in specific service content and fee standards,clear regulatory policy support,and a unified evaluation system.Significant differences exist among hospitals of different levels and types in terms of their capacity to provide pharmaceutical services and the forms in which they are offered.There is inconsistency among pharmacists within hospitals regarding crucial aspects of establishing pharmaceutical service fees,and further enhancement is needed in their awareness of relevant policies and the latest guidelines.Conclusions There is considerable room for improvement in establishing pharmaceutical service fees in hospitals of Shaanxi province.Stakeholders should promptly establish and standardize the fee establishment model,differentiate the fee standards for various services,enhance the publicity and dissemination of relevant document requirements to support the smooth implementation of pharmaceutical service fee policies.

Objective:To investigate the effect of integrated psychological and behavioral intervention on cognitive function and P300 in children with attention-deficit/hyperactivity disorder (ADHD).Methods:A randomized controlled study was conducted on 112 children with ADHD admitted to the Shaoxing 7 th People's Hospital from March 2022 to September 2023. These children were randomly divided into two groups: Group A ( n = 56) and Group B ( n = 56), using a random number table method. Group A received conventional drug treatment, whereas Group B received conventional drug treatment and an integrated psychological and behavioral intervention. The treatment lasted for 3 months. Before and after treatment, the levels of norepinephrine (NE) and dopamine (DA), scores on the Integrated Visual and Auditory Continuous Performance Test, electroencephalogram θ/β ratio, and P300 (latency and amplitude) were determined in each group. The clinical efficacy of the treatment was evaluated in both groups. Results:After treatment, the NE and DA levels in Group B were (340.83 ± 43.22) ng/L and (1.12 ± 0.25) ng/L, respectively. These levels were significantly lower than those in Group A [(416.15 ± 48.37) ng/L, (1.30 ± 0.31) ng/L, t = 8.69, 3.38, both P < 0.05]. The full scale attention quotient and full scale response control quotient in Group B were (82.52 ± 14.15) points and (96.34 ± 13.95) points, respectively. These values were significantly higher than those in Group A [(75.20 ± 13.83) points, (90.62 ± 15.16) points, t = -2.77, -2.08, both P < 0.05). The electroencephalogram θ/β ratio was significantly lower in Group B than in Group A ( t = 3.22, P < 0.05). The latencies of P300 in the Cz and Fz regions in Group B were (346.21 ± 40.52) ms and (344.65 ± 39.52) ms, respectively. The latencies were significantly shorter than those in Group A [(376.16 ± 43.04) ms, (369.35 ± 42.16) ms, t = 3.79, 3.20, both P < 0.05]. Similarly, the amplitudes of P300 in the Cz and Fz regions were significantly greater in Group B [(7.73 ± 1.80) μV, (7.22 ± 1.73) μV] than in Group A [(5.90 ± 1.15) μV and (5.76 ± 1.12) μV] ( t = -6.41, -5.30, both P < 0.05). The total response rate in Group B was significantly higher than that in Group A [94.6% (53/56) vs. 82.1% (46/56), χ2 = 4.26, P < 0.05]. Conclusion:The integrated psychological and behavioral intervention in children with ADHD is good. It can reduce NE and DA levels, improve cognitive function, decrease electroencephalogram θ/β , and improve P300.

Objective:To investigate the association of serum fibroblast growth factor 23(FGF23) level with insulin resistance(IR) and sex hormone levels in patients with polycystic ovary syndrome(PCOS).Methods:A retrospective study was performed in eighty-seven patients with PCOS, fifty-seven patients with simple IR, and sixty-one healthy women who were admitted to Affiliated Hospital of Zunyi Medical University during October 2021 and November 2022. According to the homeostasis model assessment-IR index, all subjects were divided into normal control group( n=61), IR group( n=57), PCOS without IR group(PCOS group, n=15), and PCOS+ IR group( n=72). The levels of serum FGF23, adiponectin, and sex hormones in all groups were compared, and their correlations with glucose and lipid metabolism indicators were analyzed. Results:The FGF23 level was significantly elevated in the IR group, while markedly reduced in the PCOS group and PCOS+ IR group, with the PCOS group showing a significantly lower concentration. The adiponectin levels were significantly decreased in the IR group, PCOS group, and PCOS+ IR group(all P<0.05). The correlation analysis showed that FGF23 level was positively correlated with adiponectin and sex hormone binding globulin, and negatively correlated with luteinizing hormone, luteinizing hormone/follicle stimulating hormone, and free testosterone index(all P<0.05). Logistic regression results indicated that both FGF23 and adiponectin could be used as good indicators for the diagnosis of PCOS and PCOS with IR(all P<0.05). Conclusion:FGF23 is closely related to IR and androgen as well, and under certain conditions, it can reflect the severity of IR and hyperandrogenemia in PCOS patients. The cutoff value of FGF23 obtained in this study can provide a good reference for the diagnosis of PCOS diseases.

Objective:To evaluate the efficacy and safety of Tuina(Chinese therapeutic massage)in the treatment of functional dyspepsia(FD)in children and adults. Methods:Related articles in PubMed,Excerpta Medica Database(EMBASE),Cochrane Library,Web of Science,China Biology Medicine Disc(CBM),Wanfang Academic Journal Full-text Database(Wanfang),China National Knowledge Infrastructure(CNKI),and Chongqing VIP Database(CQVIP)were collected.The retrieval time was from each database's start to March 2022.Two researchers independently screened the literature,extracted the data,and evaluated the risk of bias in the included studies.A meta-analysis was then performed using the RevMan 5.4 software. Results:A total of 19 clinical trials were included,9 of which encompassed studies on adults while 10 were on children with FD,comprising a total of 1961 patients.The findings of the meta-analysis showed that the effective rate of FD in children and adults treated with Tuina was significantly higher than that in the control group[risk ratio(RR)=1.15,95%confidence interval(CI)(1.09,1.21),P<0.001],[RR=1.13,95%CI(1.06,1.21),P<0.001].In addition,the effective rate of FD in children and adults treated with Tuina combined with other treatments was significantly higher than that in the control group[RR=1.14,95%CI(1.07,1.21),P<0.001],[RR=1.12,95%CI(1.02,1.24),P=0.02].In terms of single symptoms,Tuina improved epigastric burning sensation score in adults[standardized mean difference(SMD)=-0.41;95%CI(-0.79,-0.02);Z=2.08;P=0.04]compared with that of the Western medicine group.Compared with children treated with oral Chinese medications(CM)or Chinese patent medicine(CPM),children with FD demonstrated lower scores of epigastric pain[SMD=-0.38,95%CI(-0.56,-0.19);Z=3.96;P<0.001],postprandial fullness[SMD=-0.30,95%CI(-0.50,-0.10);Z=2.88;P=0.004],and early satiety[SMD=-0.26,95%CI(-0.47,-0.06);Z=2.54;P=0.01]after receiving Tuina combined with CM or CPM treatment.No adverse events were reported in the Tuina treatment group,and the follow-up indicated that the symptom scores in the Tuina group improved. Conclusion:Compared with the control group,both Tuina and Tuina combined with other treatments are shown to have better effective rates,lower incidence of adverse events,and better follow-up outcomes.The study results suggest that Tuina may be a clinically viable complementary therapy.However,due to limitations in the number and quality of the included studies,the above conclusions should be verified by further high-quality studies.

Objective: To explore the changing characteristics of Wnt2/β⁃catenin pathway in liver regeneration and repair in C57BL/6 mice. Methods : Male C57BL/6 mice were randomly divided into a sham⁃operated group (Sham group) , 1 d post⁃operative group , 2 d post⁃operative group , 4 d post⁃operative group , 6 d post⁃operative group , and 8 d post⁃operative group. Mice in the operated group underwent partial hepatectomy ( PHx) to remove the left and middle lobes of the liver, respectively. Plasma and liver tissues were collected on postoperative days 1 , 2 ,4 ,6 , and 8 , and plasma ALT and AST activity was measured by ALT(alanine aminotransferase) and AST( aspartate aminotransferase) biochemical analysis kits; Ki67 ⁃positive cells were identified by immunohistochemistry in each group;the number of HNF4⁃α and Ki67 double positive cells , the number of LYVE1 and Ki67 double positive cells , and the number of β ⁃catenin transferred into the nucleus were determined by immunofluorescence method. The expression of Wnt2 protein in each group was detected by Western blot , and the time characteristics of its expression during liver regeneration were analyzed. Results: Liver weight and liver/body weight ratio peaked on day 6 after PHx and approached the level of the Sham group. After PHx , there was severe liver damage on day 1 , but it had normalized by day 4. On day 2 after PHx , mainly hepatocytes proliferated;and on days 4 and 6 , mainly liver sinusoidal endothelial cells proliferated ,while the Wnt2/β⁃catenin pathway was activated. Conclusion The liver has a powerful regenerative repair ability ,which is closely related to the rapid proliferation of hepatocytes and liver sinusoidal endothelial cells , and the activation of the Wnt2/β⁃catenin pathway is activated in the regenerative repair of the mouse liver.

Objective:To investigate the serum and follicular fluid levels of sterol regulatory element-binding protein 1c(SREBP-1c), leucine-rich α-2-glycoprotein 1(LRG1) and the correlation with insulin resistance(IR) in non-ovarian etiology infertility patients and polycystic ovary syndrome(PCOS) patients with or without IR.Methods:Forty-nine PCOS patients and 66 infertility patients with non-ovarian etiology were collected in this retrospective study, homeostasis model assessment for insulin resistance(HOMA-IR) was used to evaluate IR, and were divided into control group( n=36), IR group( n=30), PCOS alone group( n=28) and PCOS-IR group(PCOS with IR group, n=21). The concentrations of serum, follicular fluid LRG1 and SREBP1c levels in each group were compared, and their correlation with relevant hormones and glycolipid metabolism were analyzed. Results:The levels of serum, follicular fluid LRG1 and SREBP1c in IR group, PCOS alone group and PCOS-IR group were significantly higher than those in control group; The PCOS-IR group showed a more significant increase in the levels of serum, follicular fluid LRG1 and SREBP1c( P<0.05). Correlation analysis showed that serum, follicular fluid LRG1 was positively correlated with body mass index, fasting plasma glucose(FPG), fasting insulin(FINS), triglycerides(TG), and HOMA-IR( P<0.05). Serum, follicular fluid SREBP1c was positively correlated with body mass index, FPG, FINS, TG, total cholesterol, LDL-C, LH, total teststerone, DHEAS, FAI, and HOMA-IR( P<0.05). Binary logistic regression analysis showed that serum SREBP1c was a risk factor for PCOS( P<0.05). Conclusion:The serum and follicular fluid levels of LRG1 and SREBP-1c were elevated in PCOS patients, especially in those with IR. The elevated levels of serum and follicular fluid LRG1 and SREBP-1c may be associated with IR and glucose-lipid metabolism abnormalities in PCOS patients. Serum LRG1 and SREBP-1c levels may serve as new indicators for predicting IR, early diagnosis, and intervention in PCOS patients.

ObjectiveTo present the health status of traditional Chinese medicine （TCM） constitutions more intuitively and comprehensively based on improved radar chart. MethodsParticipants who completed a 26-week comprehensive intervention based on TCM constitution from February 2013 to January 2014 in Zhuhai branch of Guangdong Provincial Hospital of Chinese Medicine were included in the study. They were divided into groups according to gender and age， i.e. young， middle-aged， and elderly male and female groups. TCM constitution scale and health survey short form （SF-36） were used to evaluate the 9 basic TCM constitution types and quality of life at three time points， including pre-intervention （T1）， at 13-week intervention （T2）， and at 26-week intervention （T3）. The improved radar charts were drawn to visually present the comprehensive evaluation results on the health status of 9 TCM constitutions， and graphic features （area S value， perimeter L value） were extracted to construct a comprehensive health index for TCM constitutions （H value）. Spearman correlation analysis was used to analyze the correlation between H value and SF-36 total score. ResultsAmong the included 509 participants， there were 45 elderly male， 76 elderly female， 60 middle-aged male， 140 middle-aged female， 53 young male and 135 young female. The radar charts for comprehensive evaluation of TCM constitution health status showed that the total areas for all groups increased at T3 compared to T1， with the most significant increase in the young population. In the middle-aged population， the fan-shaped areas of certain constitutions decreased at T2 than T1. At T3， the radar chart shapes for females were more balanced than males in the same age group. By calculating the features of function graphs， it was found that the S， L， and H values for the elderly population were relatively higher than those for the middle-aged and young population with the same gender， and the young population increased by highest ratio. The values measured at T3 compared to T1 showed average increase of 26% for S value （11% for the middle-aged and 14% for the elderly）， 22% for L value （10% for the middle-aged and the elderly each）， and 22% for H value （10% for the middle-aged and 9% for the elderly）. The female had lower S and L values， as well as higher H value than the male of the same age group measured at T3. The correlation coefficient between the H value of all participants and the total SF-36 score was 0.662 （P＜0.01）. ConclusionThe comprehensive evaluation model for the health status of TCM constitution based on the improved radar chart constructed in this study can present the health status of TCM constitutions and intervention effectiveness more comprehensively and intuitively. It is suggested to regulate the constitution in pursuit of the dynamic balance of the constitution health status， as well as consider the parts from the whole， and put focus on the balance of nine TCM constitutions.

Objective:To construct the evaluation index system of the core competence of neonatal specialist nurses, so as to provide reference for clinical training of neonatal specialist nurses.Methods:From January 2020, through literature review, theoretical analysis, interview, Delphi method and superiority chart, the evaluation index and weight of core competence of neonatal specialist nurses were determined.Results:Totally 28 experts in China were invited for 3 rounds of consultation. The effective recovery rate of the questionnaires was 93.33%(28/30) in the first round. The effective recovery rate of the questionnaires was 100.00%(28/28) in the second and third rounds. The authority coefficient of the experts was 0.85 in the third round. The Kendall′s coefficients of concordance of the first-level indexes, second-level indexes and third-level indexes were 0.150, 0.221 and 0.161, respectively. The final evaluation index system of the core competence of neonatal specialist nurses included 5 first-level indicators, 17 second-level indicators and 58 third-level indicators.Conclusions:The evaluation index system of the core competence of neonatal specialist nurses constructed in this study has certain scientific, reliable and clinical application value, which is conducive to the training of newborn specialized nurses in China.