Guided by the theory of "mutual dependence of ascending and descending"， recurrent pediatric cough and asthma are considered to result from the imbalance of the ascending and descending of zang-fu organ qi， which is closely associated with deficiency， phlegm， and blood stasis. In clinical practice， the disease is treated based on differentiation during the initial stage， progression stage， and stable stage. In the initial stage， the pathogenesis is attributed to lung deficiency complicated by external pathogens and liver qi rising to attack the lung； the treatment should focus on restraining the lung and calming the liver， using the self-formulated Longchai Yuchuan Fomulation （龙柴愈喘方）. During the progression stage， the disorder is due to the spleen failing to ascend clear qi and the kidney failing to grasp qi， leading to phlegm formation from deficiency； treatment should focus on tonifying spleen yang， supplementing kidney qi， and resolving phlegm and fluid retention， using a modified combination of Linggui Zhugan Decoction （苓桂术甘汤） and Jinkui Shenqi Pill （金匮肾气丸）. In the stable stage， qi deficiency leads to poor consolidation， with phlegm turbidity and blood stasis； the treatment should aim at tonifying qi， transforming phlegm， and dispelling blood stasis， using a modified version of Guizhi Fuling Pill （桂枝茯苓丸）.

Based on the "sweat pore-qi and liquid-collaterals" theory， it is considered that the core pathogenesis of pediatric infectious mononucleosis lies in the obstruction of sweat pores， the failure of qi and liquid to disperse， and damage to the collaterals due to pathogenic toxins. Accordingly， the treatment principles proposed include unblocking the sweat pores， regulating qi and liquid， and smoothing the collaterals. In clinical practice， treatment is differentiated according to stages： initial， acute， and late stages. In the initial stage， invasion of warm pathogenic toxins into the lung defense leads to obstruction of the sweat pores， which should be treated by unblocking the sweat pores and expelling pathogens outward. In the acute stage， the obstruction of the sweat pores worsens， leading to the failure of qi and liquid dispersal， resulting in intense heat toxins with accumulation of dampness， phlegm， and blood stasis， which should be treated by promoting qi movement， resolving dampness and phlegm， clearing heat， detoxifying， and dispersing stasis to regulate qi and liquid. In the late stage， residual pathogens remain， with qi and yin deficiency and unsmooth collaterals， which should be treated by unblocking the collaterals， dissipating nodules， tonifying qi， and nourishing yin to smooth the collaterals. This approach may provide new insights for the clinical treatment of pediatric infectious mononucleosis.

IKBKG is the essential modulator for nuclear factor-κB(NF-κB) signaling pathway, and mutations within this gene can lead to anhidrotic ectodermal dysplasia and immunodeficiency (EDA-ID). Here we report a male patient, who presented with mild frontal bossing, sparse hair, skin pigmentation, conical teeth, and recurrent infections involving bacteria, fungi, and viruses after one month of age, together with hypogammaglobulinemia. These symptoms were consistent with the phenotype of EDA-ID. Genetic analysis revealed a hemizygous mutation c.1249T > G (p.Cys417Gly) in exon 10 of the IKBKG gene in the patient. The mother of the patient was identified as heterozygous carriers of the same mutation. Immunological assessment revealed a significant reduction in memory B cells. The patient showed no skewed TCR diversity. PHA-induced T-cell proliferation was impaired. IFN-γ secretion by Th cells was significantly reduced after PHA stimulation. IκBα degradation rate retained the same in the patient. We summarized the clinical features of the patient and conducted immunological analysis, in order to increase pediatricians′awareness of this rare disease. For boys with early-onset recurrent infections together with ectodermal dysplasia, IKBKG mutation should be considered. In addition, genetic analysis is needed to exclude pseudogene interference and functional evaluations are required.

Objective:To investigate the effectiveness and safety of low-dose oral misoprostol solution for cervical ripening in late gestation.Methods:This was a prospective cohort study including 396 primiparas with singleton pregnancy who received low-dose oral misoprostol solution for cervical ripening (oral group) in Peking University Third Hospital from March to October 2022. They were further allocated to receive oral misoprostol alone (OA group, n=167) or oral misoprostol in combination with oxytocin/amniotomy (OC group, n=229). Moreover, 218 cases who received vaginal misoprostol for cervical ripening (vaginal group) during the same period in 2021 were reviewed (a retrospective cohort). Among them, 77 were given vaginal misoprostol alone (VA group) and 141 received vaginal misoprostol in combination with oxytocin/amniotomy (VC group). The OA group and VA group (72 and 73 cases) as well as the OC group and VC group (108 and 103 cases) were matched using propensity scores. Basic clinical information, hospital stay, duration of labor induction, uterine hyperstimulation, rate of labor initiation, vaginal delivery rate, rate of delivery within 24 h, duration of labor, neonatal condition, adverse pregnancy outcomes, and other information were compared between different groups. All data were statistically analyzed using independent sample t test, analysis of variance, nonparametric test, Chi-square test, or Fisher's exact probability test. Logistic regression model was used to analyze the factors affecting the labor initiation and the failure of labor induction. Results:The average hospital stay, the duration from medication to labor initiation and the duration from medication to vaginal delivery were significantly shorter in the oral group than those in the vaginal group [(5.4±2.4) vs. (6.5±2.6) d, (34.2±24.1) vs. (38.9±25.7) h, (45.8±25.8) vs. (53.4±27.8) h; t=5.24, 2.10 and 3.39; all P<0.05]. The total labor initiation rate and vaginal delivery rate in the oral group were significantly higher than those in the vaginal group [92.9% (368/396) vs. 83.5% (182/218), 72.2% (286/396) vs. 60.1% (131/218); χ 2=13.43 and 9.50; both P<0.05]. The incidence of failed induction of labor, uterine hyperstimulation, fetal distress, and intrauterine infection in the oral group were lower than those in the vaginal group [2.0% (8/396) vs. 6.9% (15/218), 4.3% (17/396) vs. 17.9% (39/218), 8.8% (35/396) vs. 14.7% (32/218), 1.3% (5/396) vs. 3.7% (8/218); χ 2=9.21, 31.36, 4.93 and 3.93; all P<0.05]. The duration from medication to labor initiation and to vaginal delivery in the OA group were higher than those in the VA group [(25.8±17.0) vs. (17.4±10.8) h, (37.2±18.8) vs. (29.7±13.5) h; t=3.49 and 2.74; both P<0.05]. There were no significant differences in the labor initiation rate, vaginal delivery rate, rate of delivery within 24 h or the incidence of failed induction of labor between the OA and VA groups (all P>0.05). Women in the VA group were more likely to develop uterine hyperstimulation than those in the OA group [19.2% (14/73) vs. 4.2% (3/72), χ2=7.89, P=0.005]. There were no significant differences in the duration from medication to labor initiation or to vaginal delivery between the VC and OC groups (both P>0.05), but the duration were significantly longer than those in the corresponding medication alone group (VC vs. VA groups: (49.7±24.6) vs. (17.4±10.8) h and (61.6±25.7) vs. (29.7±13.5) h, t=5.31 and 5.13, both P<0.05; OC vs. OA groups: (45.3±26.6) vs. (25.8±17.0) h and (56.1±27.2) vs. (37.2±18.8) h, t=10.35 and 9.78, both P<0.05]. The labor initiation rate, vaginal delivery rate and rate of delivery within 24 h in the OC group were higher than those in the VC group [88.9% (96/108) vs. 77% (87/113), 63.0% (68/108) vs. 47.8% (54/113), 10.3% (7/108) vs. 0.0% (0/113); χ 2=5.49, 5.14 and 7.56; all P<0.05]. The incidence of uterine hyperstimulation in the OC group was 4.6% (5/108), which was lower than that in the VC group [18.6% (21/113), χ 2=10.37, P=0.001]. Logistic regression analysis showed that oral misoprostol and gestational age were positively correlated with labor initiation [ OR (95% CI): 2.18 (1.24-3.90) and 1.43 (1.14-1.79)], while maternal age was negatively correlated with labor initiation [ OR (95% CI): 0.90 (0.82-0.98)]. Moreover, failed induction of labor was negatively correlated with oral misoprostol [ OR (95% CI): 0.37 (0.14-0.91)], but positively correlated with maternal age [ OR (95% CI): 1.21 (1.05-1.40)]. Conclusions:Oral administration of low-dose misoprostol solution is as effective as vaginal misoprostol in promoting cervical ripening. Besides, it can shorten the average hospital stay and reduce the incidence of uterine hyperstimulation, suggesting that low-dose oral misoprostol solution is relatively safer and can be used to promote cervical ripening in late gestation.

Objective To develop a multiplex PCR assay for simultaneous detection of four intestinal parasites, including Giardia duodenalis, Cryptosporidium parvum, Enterocytozoon bieneusi and Moniezia, and to preliminarily evaluate its detection efficiency. Methods Four pairs of specific primers were designed based on the conserved sequences of the corresponding genes of G. duodenalis (GenBank accession number: XM_001710026.2), C. parvum (GenBank accession number: XM_626998.1), E. bieneusi (GenBank accession number: KJ719492.1) and Moniezia (GenBank accession number: OM296991.1) retrieved from the GenBank database, and a multiplex PCR assay for simultaneous detection of G. duodenalis, C. parvum, E. bieneusi and Moniezia was developed and optimized. A total of 116 fresh goat stool samples were collected from four goat farms in Zhanjiang City, Guangdong Province during the period from October to December 2022, including 96 samples used for evaluating the detection efficacy of the multiplex PCR assay, and 20 samples as baseline controls for sample testing. Genomic DNA extracted from 96 goat stool samples was tested using the single-target PCR assay and the developed multiplex PCR assay, and the sensitivity, specificity, positive predictive value, and negative predictive value of the multiplex PCR assay were evaluated for detection of G. duodenalis, C. parvum, E. bieneusi and Moniezia DNA in goat stool samples with the single-target PCR assay as the gold standard. Results The multiplex PCR assay developed in this study allowed simultaneous amplification of specific gene fragments of G. duodenalis, C. parvum, E. bieneusi and Moniezia, with 1 400, 755, 314 bp and 585 bp in sizes, respectively, and the detection limit was 10² and higher copies of parasite DNA clones, while the multiplex PCR assay was negative for gene amplification of Schistosoma japonicum, Fasciola hepatica, Echinococcus granulosus, Blastocystis hominis and Homalogaster paloniae. Single-target PCR assay and the developed multiplex PCR assay were employed to test DNA samples extracted from 96 goat stool samples, and single-target PCR assay tested positive in 40 goat stool samples (41.67%), including 39 positive samples tested with the multiplex PCR assay, with a mean coincidence rate of 97.50% (39/40). The multiplex PCR assay tested positive for G. duodenalis DNA in 26 goat stool samples (27.10%), C. parvum DNA in 22 samples (22.90%), E. bieneusi DNA in 24 samples (25.00%), and Moniezia in 9 samples (9.40%), which was consistent with the detection using the single-target PCR assay. The sensitivity, negative predictive value, and positive predictive value of the multiplex PCR assay were 96.15%, 95.83%, 100.00% and 100.00%, 98.90%, 98.92%, 100.00% and 100.00%, 100.00%, 100.00%, 100.00% and 100.00% for detection of G. duodenalis, C. parvum, E. bieneusi and Moniezia DNA in goat stool samples, respectively, if the single-target PCR assay served as the gold standard. Conclusion A highly sensitive and specific multiplex PCR assay has been developed for simultaneous detection of G. duodenalis, C. parvum, E. bieneusi and Moniezia in goats, which is suitable for rapid, large-scale screening of intestinal parasites in sheep stool samples.

Objective The study aims to optimize the conditions for constructing orthotopic nude mouse models of liver cancer by injecting human liver tumor cell lines and to explore appropriate timings for drug administration. Methods Human hepatocellular carcinoma Hep3B and hepatoblastoma HepG2 cell lines, which stably expressing the luciferase reporter gene (LUC), were selected. The linear correlation between the luciferase luminescence intensity and the number of liver tumor cells was analyzed using a Small Animal In Vivo Imaging system to verify the luminescent efficiency of the human liver tumor cells. Different concentrations (8×10⁶, 2.4×10⁷, 7.2×10⁷ cells/mL) and resuspension media (PBS, Matrigel) of human liver tumor cell suspensions HepG2-LUC and Hep3B-LUC were orthotopically inoculated into the liver lobes of 5-week-old female BALB/c nude mice (12 groups, 7 mice each) to construct human liver tumor nude mouse orthotopic cancer models. Every 7 days, the weights of mice were recorded, and the growth of orthotopic tumors was monitored using the Small Animal In Vivo Imaging system. On day 35 post-cell inoculation, mouse livers were dissected, and pathological slices were prepared for HE staining to observe histopathological changes in liver tissues. Results The luminescence intensity of human liver tumor cell lines was positively correlated with the number of cells (R²=0.983 1, R²=0.970 5), indicating their suitability for orthotopic model construction. Successful modeling was achieved in the high-concentration groups of HepG2-LUC, the low-, medium-, and high-concentration groups of HepG2-LUC+Matrigel, the medium- and high-concentration groups of Hep3B-LUC, and the low-, medium-, and high-concentration groups of Hep3B-LUC+Matrigel. For both HepG2-LUC+Matrigel and Hep3B-LUC+Matrigel groups, mice in the high-concentration groups exhibited significantly reduced body weight compared to the low- and medium-concentration groups (both with P<0.05). The luminescence intensity of successfully modeled mice increased exponentially over time (R²>0.950 0), and reached a minimum of 1.0×10⁷ p/(s·cm²·sr) by day 14 post-transplantation. Mice in the low- and medium-concentration groups of HepG2-LUC and the low-concentration group of Hep3B-LUC showed no significant pathological changes, while the other groups exhibited evident liver tumors and hepatocyte lesions. Conclusion For the HepG2-LUC cell line, the recommended injection volume is 50 µL with a cell density of 2.4×10⁷ cells/mL, resuspended with Matrigel, followed by drug administration or prognostic measures on day 7 post-modeling. For the Hep3B-LUC cell line, the recommended injection volume is 50 µL with a cell density of 7.2×10⁷ cells/mL, not resuspended with Matrigel, with administration or prognostic measures on day 14 post-modeling.

[Objective] To establish a real-time fluorescence quantitative PCR nucleic acid detection method of human parvovirus B19 and validate the method systematically. [Methods] Specific primers and probes for the highly conserved regions of the three genotypes of B19 virus were designed, and B19 quantitative amplification standard curves were established. The accuracy, precision (repeatability and intermediate precision), linear range, quantification limit, detection limit, specificity, anti cross contamination, genotyping and anti-interference ability of this method were verified. [Results] When the quantitative reference range for B19 virus was 2.0×10¹ to 1.0×10⁸ IU/mL, a double logarithmic regression analysis was performed between the measured values and the theoretical values, and the regression equation R²≥0.98 showed good linear correlation. The quantification limit was 20 IU/mL, with a detection rate of 100%. The detection limit was 10 IU/mL, and the detection rate is 95.23%. Three genotypes of B19 virus samples can be effectively detected. The plasma of seven non B19 pathogens, including hepatitis A virus, hepatitis B virus, hepatitis C virus, human immuno-deficiency virus, human cytomegalovirus, hepatitis E virus and Treponema pallidum, was non reactive and has good species specificity. Simultaneously, in the presence of seven other concurrent pathogens, positive samples with a weak positive concentration of E3 IU/mL could be stably detected, and the B19 nucleic acid testing method was not interfered with. When the hemoglobin concentration was 431 mg/dL, triglycerides (1 269 turbidity) and unconjugated bilirubin concentration was 20 mg/dL, this method was non reactive for all three common plasma interfering substances. In the presence of three common plasma interfering substances, positive samples with a weak positive concentration of E3 IU/mL could be stably detected, and the B19 nucleic acid testing method was not interfered with. The deviation between the detection values of standard substances at two concentration levels of S1 (E5 IU/mL) and S2 (E4 IU/mL) and the target values were≤±0.5 log value. The CV values of positive sample 1 (concentration level E5 IU/mL) and positive sample 2 (concentration level E4 IU/mL) for daily precision confirmation and continuous 5-day intra-day precision confirmation were both≤5%. [Conclusion] This method has strong specificity, high sensitivity, wide linear range, stability, reliability and high accuracy, and can be used for the detection of human parvovirus B19 nucleic acid in plasma.

OBJECTIVE To study the anti-ulcerative colitis(UC) effect of Sargentodoxae Caulis and explore its mechanism. METHODS The UC mice model induced by dextran sodium sulfate was used to evaluate the anti-UC effect of Sargentodoxae Caulis. The ingredients of Sargentodoxae Caulis were obtained according to the CNKI and PubMed website, component targets were screened by SwissTargetPrediction database, GEO gene chip was used to extract UC differential genes, then a network of "ingredients-targets-disease" of the Sargentodoxae Caulis was constructed. After screening the core targets, protein interaction and cluster analysis, biological process and pathway enrichment analysis were performed, and the reliability of network analysis was preliminarily verified by molecular docking and literatures. RESULTS Sargentodoxae Caulis could significantly improve the disease activity index score, colon shortening and colonic histopathological changes of UC mice, and had a good anti-UC effect. The network analysis found that the core components of the anti-UC of Sargentodoxae Caulis include (+)-Dihydroxyurearetic acid, Isorhaponigenin and Pinosylvin, and 63 core targets, such as EGFR, STAT1 and LCK, regulating PI3K-Akt signal pathway and cancer proteoglycan and other related signal pathways of immune anti-inflammatory and anti-cancer, and it could affect the biological processes such as amino acid modification, kinase activity regulation, cell reaction and oxidative stress to treat UC. Molecular docking and literature showed that the constructed network had high reliability. CONCLUSION Sargentodoxae Caulis has a good anti-UC effect, and its mechanism may be closely related to the regulation of intestinal immune inflammation and cell proliferation, differentiation and migration. It has the characteristics of multi-component, multi-target and multi-way.

Objective To investigate the effect of cinobufacini on inhibiting colorectal cancer metastasis by regula-ting the polarization of M2 macrophages.Methods THP-1 was induced into M0 type macrophages.The condi-tioned medium of HCT116 cells was collected to stimulate M0 type macrophages.The polarization of M2 type mac-rophages was observed by flow cytometry,real-time quantitative PCR and ELISA experiments.The conditioned me-dium of M0 type macrophages and HCT116-Mφ cells was collected to stimulate HCT116 cells.The ability of migra-tion and invasion was observed by wound healing assay and Transwell assay.The effect of cinobufacini on the via-bility of HCT116 cells was detected by CCK-8 assay.The conditioned medium of HCT116 and HCT116+cinobufa-cini was collected to stimulate M0 type macrophages.The polarization of M2 type macrophages was observed by flow cytometry,real-time quantitative PCR and ELISA experiments.The conditioned media of HCT116-Mφ cells and(HCT116+cinobufacini)-Mφ cells were collected to stimulate HCT116 cells.The changes of migration and inva-sion ability were observed by wound healing assay and Transwell assay.Results After stimulation of M0 type mac-rophages in HCT116 cell conditioned medium,the morphology of M0 macrophages turned into fusiform cells,the proportion of CD11b+CD206+cells increased,and the expression of M2 macrophage markers IL-10 and TGF-β in-creased.The migration and invasion ability of HCT116 cells were significantly enhanced after stimulation in the conditioned medium of HCT1 16-Mφ cells.After the addition of cinobufacini,not only the polarization proportion of M2 macrophages decreased,but also the metastatic effect mediated by M2 macrophages was inhibited.Conclusion HCT116 cells can induce the polarization of M2 macrophages,while cinobufacini can inhibit the tumor metastasis mediated by M2 macrophages by inhibiting the polarization of M2 macrophages.

Objective To explore the molecular mechanism of Atractylodes macrocephala in the treatment of Ulcerative colitis(UC)based on network pharmacology,and verify it with animal experiments.Methods The active components of Atractylodes macrocephala was screened from the TCMSP database,the TCM-ID database,and in combination with relevant references,and the corresponding targets were obtained through Swiss database.The relevant targets of UC were obtained from GeneCards database,construct the"drug-component-target-disease"network diagram and"pathway-active ingredient-target"network diagram and draw PPI network diagram;GO function enrichment analysis and KEGG signal pathway annotation analysis were carried out.Autodock software is used for molecular docking of active components and targets.Then,the experimental validation of the network pharmacology prediction was carried out.The mouse UC model was induced by dextran sodium sulfate(DSS).The pathological changes of the colon tissue,the number of goblet cells,and the positive expression of inflammatory factorswere detected by HE staining,AB-PAS staining and immunohistochemistry in colon tissue of UC mice.Results The results have shown 30 active ingredients including atractylolactone I,II and III were screened,and 591 corresponding targets were obtained,of which the key target was IL-1β、TNF-α and so on.Molecular docking show that the core components had good binding affinity with the key targets.And the results of animal experiments showed that the alcohol extract of Atractylodes macrocephala could significantly increase the colon length,reduce the DAI score,improve the pathological changes of colon tissue of UC mice,increase the number of goblet cells,and inhibit the expression of IL-1β,TNF-α in colon tissue.Conclusion This study indicated that Atractylodes macrocephala could regulate the release of inflammatory factors through multiple components,multi-target and multi-channel,which could inhibit inflammatory reaction and play a role in improving UC.