AIM: To provide references for the non-clinical evaluation of therapeutic targets or drugs for retinoblastoma, fluorescently labeled Y79 cells are injected into the vitreous body of BALB/c-nu mice to establish a retinoblastoma model, and the Melphalan treatment group is used as a positive control, which is verified by fluorescence imaging technology.METHODS: BALB/c-nu mice were intravitreous injected with GFP transfected Y79 cells(1.0×107 cell/mL, 3 μL)to establish the model. On the 27th day, the mice were randomly divided into model control group and different doses of Melphalan groups(1, 3, 10 μg/eye groups)according to the fluorescence value of in vivo imaging, with vitreous body single administrated and ocular symptoms observed daily. Slit-lamp examination was performed at 12, 20, 29, 35, 42, 48, 55, 76, and 83 d after modeling. In vivo imaging was performed on 12, 20, 27, 41, 48, 55, 62, 69, 76, and 83 d. At the last treatment, the eyeball, brain and cerebellum tissues were removed for histopathological examination.RESULTS: From the sixth day of modeling, cloud-like substances could be seen in the eyes of the animals, and the cloud-like substances occupied the whole eyeball of the mice in the model control group at the later stage, accompanied by irregular growth of blood vessels. After 27 days of modeling, the fluorescence value was detected in all the animals, and the fluorescence value continued to increase with the extension of modeling time. The fluorescence value of the tumor reached the peak after 69-83 days of modeling. Histological examination showed severe proliferation of intraocular tumor cells in the model control group, and tumor cells were observed in the brain of 1 model animal. In the 10 μg/eye Melphalan group, the fluorescence value was significantly decreased at 17 d after administration. The fluorescence value of the 3 μg/eye Melphalan group was significantly inhibited at 59 d after administration. No tumor cells were found in the brain tissue of animals in all Melphalan groups.CONCLUSION: After vitreous injection of Y79/pCDH-LUC-copGFP cells in BALB/c-nu mice, significant ocular lesions and proliferation of tumor cells were observed in the eyes. Meanwhile, Melphalan intervention significantly inhibited tumor cells in a dose-dependent manner, indicating that the mouse model of retinoblastoma was successfully constructed.

Objective:To establish the reference intervals (RIs) of systemic immune inflammatory index (SII), platelet to lymphocyte ratio (PLR), neutrophil to lymphocyte ratio (NLR), lymphocyte to monocyte ratio (LMR) and monocyte to lymphocyte ratio (MLR) in healthy pregnant women in Henan province, China.Methods:A retrospective analysis was conducted on the data of the healthy pregnant women without a history of adverse pregnancy events who participated in health check-ups from August 2016 to February 2019. A total of 4 016 healthy pregnant women were selected for establishing RIs. Data from healthy adult control group were derived from the healthy adult cohort in Henan established earlier by our team, and the Propensity Score Matching analysis was used and 3 595 healthy adult women and 3 595 healthy pregnant women to compare the indicators between the two groups. The RIs of the above indicators were established using the indirect method with a 95% confidence interval. The Tukey Rule was used to identify and remove outliers. The RIs were stratified and grouped based on the differences in each indicator during the pregnancy: SII: 3 929 cases, including 712 in the first trimester, 1 947 in the second trimester, and 1, 270 in the third trimester; PLR: 3 927 cases, no grouping; NLR: 3 925 cases, including 712 in the first trimester and 3 213 in the second and third trimesters; LMR: 3 925 cases, including 723 in the first trimester, 1 942 in the second trimester, and 1 260 in the third trimester; MLR: 3 904 cases, including 721 in the first trimester, 1 928 in the second trimester, and 1 255 in the third trimester. After the RIs were established, another 396 healthy pregnant women without a history of adverse pregnancy events who participated in health check-ups from February to April 2019 were selected for the validation of the RIs.Results:SII, NLR, LMR, MLR, and PLR differ significantly between healthy adult women and healthy pregnant women. There were significant differences in SII, LMR, and MLR among the three trimesters ( P<0.05). NLR in the first trimester was significantly lower than that in the second and third trimesters ( P<0.05), while there was no significant difference between the second and third trimester ( P=0.124). PLR only showed significant differences between the second and third trimester ( P<0.05), while no significant differences were found among the other groups. Based on the above results, the stratified RIs of each index in healthy pregnant population were established and verified. SII: first trimester (341-1 426)×10 9/L, second trimester (437-1 680)×10 9/L, third trimester (379-1 580)×10 9/L; PLR: 73-215; NLR: first trimester 1.78-5.60, second and third trimester 2.21-6.74; LMR: first trimester 2.20-6.61, second trimester 1.85-5.42, third trimester 1.63-4.82; MLR: first trimester 0.14-0.42, second trimester 0.17-0.49, third trimester 0.18-0.55. The rejection rate of 396 cases was less than 10%. Conclusions:The RIs of SII, NLR, LMR, MLR and PLR for healthy pregnant women in Hernan province of China were established and validated, and4 could be used in clinical practice.

Objective:To validate and optimize the interpretation methods and cut-off values of prolonged activated partial thromboplastin time (APTT) mixing test, in order to elevate the clinical applicational value in differentiating single factor deficiency, positive lupus anticoagulant (LA) and FⅧ inhibitors.Methods:This cross-sectional retrospective study involved 130 cases with single APTT prolongation for unknown reasons who underwent mixing tests in the First Hospital of Jilin University from January 1st, 2022 to December 31st, 2023, including 84 males and 46 females. Rosner index (RI), Percentage correction ("Chang") method, Normal reference interval (NRI) method, differential analysis, post-incubation prolongation time and percentage (including mixed immediately and ) were respectively calculated and analyzed for both immediate mixing and 2-hour post-mixing incubation. According to clinical diagnosis, patients were divided into single factor deficiency (51 cases), positive lupus anticoagulant (LA) (56 cases), and positive factor Ⅷ (FⅧ) inhibitor (23 cases). Receiver operation characteristic (ROC) curve analysis was used to determine the cut-off values for APTT mixing test methods in patients with different reasons for prolonged APTT, which were evaluted and adjusted by AUC, sensitivity, and specificity, and compared with the methods currently used for interpretations.Results:For the single factor deficiency cases, the AUCs of immediate RI index <7.5% and incubated RI index <13.9% were 0.87 and 0.90, respectively, with sensitivities at 87.8% and 83.8%, and specificities at 72.0% and 80.0%, whose combination had sensitivity and specificity at 87.3%, and 94.0%, respectively; the judging accuracy increased from 84.6% (110/130) in currently used rule to 93.1% (121/130). For the positive lupus anticoagulant (LA) cases, the combination of immediate RI index >10.3% and prolonged post-incubation clotting time percentage <9.1% showed 88.5% sensitivity and 99.8% specificity; the judging accuracy improved from 80.0% (104/130) in currently used rule to 86.2% (112/130). For positive FⅧ inhibitor cases, the combination of incubated RI index >17.0% and prolonged post-incubation clotting time percentage >9.1% had 99.8% sensitivity and 100.0% specificity; the judging accuracy increased from 89.2% (116/130) in currently used rule to 94.6% (123/130).Conclusion:This study validated and optimized the interpretation methods and cut-off values for the prolonged APTT mixing test in differentiating single factor deficiencies, prositive FⅧ inhibitors and LA positive cases, significantly improving the judging sensitivity and specificity.

Objective To explore the mechanism of modified Sijunzi Decoction in the intervention of premature ovarian failure(POF)rats through PTEN-mediated PI3K/AKT signaling pathway.Methods Totally 50 SPF-grade healthy female SD rats were divided into normal group(10 rats)and model group(40 rats).The model group was injected with cyclophosphamide to establish a POF model,and then randomly divided into model group and TCM low-,medium-and high-dosage group,with 10 rats in each group,TCM low-,medium-and high-dosage group were given modified Sijunzi Decoction 6.3,12.6,25.2 g/kg by gavage,the normal group and model group were given equal volume of normal saline,once a day for 28 days.The general condition and body mass of rats in each group were observed and recorded daily.After administration,the ovarian index was measured,HE staining was used to observe the morphology of ovarian tissue,ELISA was used to detect the contents of serum estradiol(E2)luteinizing hormone(LH),follicle-stimulating hormone(FSH),anti-Müllerian hormone(AMH),gonadotropin-releasing hormone(GnRH)and β-endorphin(β-EP),RT-PCR was used to detect the mRNA expression of PTEN,PI3K and AKT in ovarian tissue,Western blot was used to detect the protein expression of PTEN,PI3K,AKT,p-PI3K and p-AKT,immunohistochemistry was used to detect the positive expression of PTEN,PI3K and AKT.Results Compared with the normal group,the rats in the model group were depressed,the hair fell off,the diet and drinking water were reduced,the stools became thinner,the body mass was reduced(P<0.05),and the ovarian index was reduced(P<0.05),the ovarian tissue structure was disordered,the number of follicles decreased,and the contents of serum LH,FSH and GnRH increased(P<0.05),the contents of E2,AMH and β-EP decreased(P<0.05),the expression of PTEN mRNA and protein decreased(P<0.05),the expressions of PI3K and AKT mRNA and protein increased(P<0.05),and the expressions of p-PI3K and p-AKT protein increased(P<0.05).Compared with the model group,the general condition of rats in each dosage of TCM groups improved significantly,the body mass increased(P<0.05),ovarian index increased(P<0.05),the number of follicles in rat ovarian tissue increased,the contents of LH,FSH and GnRH decreased(P<0.05),the contents of E2,AMH and β-EP increased(P<0.05),the expression of PTEN mRNA and protein increased(P<0.05),the expression of PI3K and AKT mRNA and protein decreased(P<0.05),and the protein expression of p-PI3K and p-AKT decreased(P<0.05).Conclusion Modified Sijunzi Decoction may regulate the PI3K/AKT signaling pathway mediated by PTEN,repair the excessive consumption of follicles in ovarian tissue of POF rats,improve the ovarian function,so as to intervene the process of POF.

Objective:To analyze the epidemiological and clinical features of infectious diseases of the central nervous system (CNS).Methods:A cross-sectional study and analysis were conducted to summarize the epidemiological and clinical characteristics of 9 918 patients with CNS infectious diseases, who were diagnosed and treated at 29 hospitals across China from January 1, 2001 to December 31, 2020. Data collected included demographic data, clinical manifestations, health economic indicators, and prognostic outcomes.Results:Among the 9 918 collected cases of CNS infectious diseases, 5 559 were male (56.0%) and 4 359 were female (44.0%), with an onset age of 38 (25, 53) years. Education level: slightly more junior high school education (2 651 cases, 26.7%), and less elementary school education and below (2 181 cases, 22.0%) were found. Occupational distribution: farmers were found predominant (3 215 cases, 32.4%), followed by workers (1 826 cases, 18.4%) and students (1 633 cases, 16.5%). Clinical manifestations: headache (6 074 cases, 61.2%), fever (5 869 cases, 59.2%) and positive meningeal irritation signs (2 273 cases, 22.9%) were the 3 most common clinical manifestations, followed by nausea and (or) vomiting (2 095 cases, 21.1%), impaired consciousness (2 077 cases, 20.9%), psychiatric symptom (1 866 cases, 18.8%) and epilepsy (1 627 cases, 16.4%), etc., and cranial nerve involvement was found in 669 cases (6.7%). Major pathogens included viruses in 6 814 cases (68.7%), Mycobacterium tuberculosis in 1 677 cases (16.9%), common bacteria in 864 cases (8.7%), fungi in 254 cases (2.6%), spirochetes of syphilis in 183 cases (1.8%), parasites in 121 cases (1.2%), and rickettsiae in 5 cases (0.1%). Urban-rural distribution: slightly more cases were found in the countryside (5 418 cases, 54.6%) than in the towns (4 500 cases, 45.4%). Distribution of onset by season: 2 412 cases (24.3%) fell ill in spring, 2 835 cases (28.6%) in summer, 2 187 cases (22.1%) in fall, and 2 484 cases (25.0%) in winter. Health economics: the duration of hospitalization was 15 (8, 27) days, and the cost of hospitalization was 1.53 (0.91, 3.02)×10 000 yuan. Prognosis: 9 531 cases (96.1%) were cured or improved, and 92 cases (0.9%) died. Conclusions:The pathogens responsible for CNS infectious diseases are predominantly viruses. Although the incidence is slightly higher during the summer months, the overall seasonal pattern is not particularly pronounced. These infections are more commonly observed in young and middle-aged males and present with a diverse range of clinical manifestations, contributing to a significant disease burden.

Objective:To investigate the risk factors for mortality in neonates with necrotizing enterocolitis (NEC).Methods:This retrospective cohort study included patients diagnosed with NEC at stage Ⅱ-Ⅲ (Bell's criteria) and admitted to the Neonatology Department of Guangzhou Women and Children's Medical Center Affiliated to Guangzhou Medical University from January 2017 to December 2022. According to the outcomes, these patients were divided into the mortality and survival groups. Perinatal conditions, clinical manifestations, disease status during hospitalization, and blood routine parameters at different time points after birth were compared between the two groups to analyze the risk factors for mortality of NEC. Statistical analysis was performed using independent-sample t-test, Mann-Whitney U test, Chi-square test (or Fisher's exact test), and multivariate logistic regression analysis. Results:(1) A total of 118 NEC cases were included, with 100 in the survival group and 18 in the mortality group. (2) The gestational age and birth weight were significantly lower in the mortality group than in the survival group [(28.9±4.2) weeks vs. (33.7±3.9) weeks, t=4.78; 940 g (685-1 275 g) vs. 1 830 g (1 352-2 368 g), Z=4.18; both P<0.05]. The incidence of neonatal asphyxia was higher in the mortality group [9/18 vs. 18% (18/100), χ2=7.13, P<0.05]. (3) Compared with the survival group, the mortality group had higher proportions of patients who were at NEC stage Ⅲ, accepted surgery, were treated with vasoactive drugs, had undergone invasive mechanical ventilation, or had metabolic acidosis, hyperlactatemia, shock, or hematochezia [17/18 vs. 24% (24/100), χ2=33.39; 17/18 vs. 31% (31/100), χ2=22.88; 16/18 vs. 22% (22/100), χ2=31.26; 16/18 vs. 39% (39/100), χ2=15.26; 18/18 vs. 28% (28/100), χ2=30.29; 16/18 vs. 20% (20/100), χ2=34.15; 17/18 vs. 21% (21/100), χ2=37.69; 9/18 vs. 82% (82/100), χ2=7.13; all P<0.05]. (4) The proportions of patients who developed late-onset sepsis, hemodynamically- significant patent ductus arteriosus, respiratory distress syndrome, or pulmonary hemorrhage were significantly higher in the mortality group than in the survival group [15/18 vs. 33% (33/100), χ2=16.01; 9/18 vs. 21% (21/100), χ2=5.32; 15/18 vs. 39% (39/100), χ2=12.08; 7/18 vs. 7% (7/100), χ2=11.94; all P<0.05]. (5) Within 24 h of birth, the levels of red blood cells and platelets were lower in the mortality group than in the survival group [4.1×1012/L (3.8×10 12/L-4.6×10 12/L) vs. 4.6×10 12/L (4.0×10 12/L-4.9×10 12/L), Z=2.04; (199.9±68.6)×10 9/L vs. (239.8±72.6)×10 9/L, t=2.16; both P<0.05]; at 6-8 d after birth, the levels of red blood cells, hemoglobin (Hb) and hematocrit (HCT) were lower in the mortality group than in the survival group [(3.2±0.5)×10 12/L vs. (3.9±0.8)×10 12/L, t=3.30; (111.2±19.2) vs. (138.1±28.3) g/L, t=3.51; (33.0±5.4)% vs. (40.9±8.1)%, t=3.61; all P<0.05]; at the diagnosis of NEC, red blood cell count, Hb level, HCT, and platelet count were lower in the mortality group than in the survival group [(3.3±1.0)×10 12/L vs. (3.8±0.8)×10 12/L, t=2.47; (102.8±28.8) vs. (124.4±26.3) g/L, t=3.59; 31.0% (25.9%-38.4%) vs. 37.2% (31.5%-43.7%), Z=2.62; 87.0×10 9/L (50.2×10 9/L-157.0×10 9/L) vs. 228.0×10 9/L (130.0×10 9/L-414.7×10 9/L), Z=3.78; all P<0.05], while mean platelet volume (MPV), platelet distribution width, and the differences in hemoglobin (ΔHb) and hematocrit (ΔHCT) between the first 24 h after birth and 6-8 d after birth were significantly higher in the mortality group than in the survival group [13.1 fl (11.4-13.6 fl) vs. 11.6 fl (10.7-12.4 fl), Z=3.26; 19.6% (13.9%-25.2%) vs. 14.8% (12.0%-18.6%), Z=2.76; 35.5 g/L (28.3-57.3 g/L) vs. 27.0 g/L (8.0-42.5 g/L), Z=2.20;11.5% (9.4%-16.3%) vs. 6.3% (2.2%-11.2%), Z=2.85; all P<0.05]. (6) Late-onset sepsis ( OR=5.568, 95% CI: 1.201-25.816), hyperlactatemia ( OR=6.702, 95% CI: 1.193-37.651), shock ( OR=10.616, 95% CI: 1.157-97.406) and MPV elevation at the diagnosis of NEC ( OR=2.769, 95% CI: 1.468-5.223) were independent risk factors, while gestational age ( OR=0.836, 95% CI: 0.708-0.986), and HCT at 6-8 d after birth ( OR=0.848, 95% CI: 0.759-0.947) were protecting fctors for death in NEC. Conclusions:Preterm infants with smaller gestational age are more prone to mortality of NEC. Early identification and management of late-onset sepsis, shock, and hyperlactatemia may reduce the risk of mortality in NEC.

Objective To explore the distribution of common TCM syndromes and symptoms of insomnia;To prepare for the construction of the theoretical framework and item pool of syndrome diagnosis and efficacy evaluation scale.Methods TCM guideline standards of insomnia,textbooks and journals over the years were retrieved,the information of TCM syndromes,syndrome elements and symptoms was extracted,the guideline textbook and journal database were established,and descriptive statistics,association rules,systematic clustering,factor analysis,potential categories and implicit structure analysis were carried out.Results Totally 116 guide standards and textbooks over the years were included,and 454 articles of journals were included.The high-frequency symptoms accounted for≥3%of the guide textbooks and journal databases were 87 and 79 categories,respectively,and the cumulative proportion was 87.48%and 87.75%,respectively.According to the analysis results,five common TCM syndromes and their characteristic symptom classification of insomnia were finally deduced.According to the frequency/person time distribution,they were heart and spleen deficiency syndrome,yin deficiency and fire hyperactivity syndrome,liver fire disturbing heart syndrome,phlegm heat disturbing heart syndrome,heart and gallbladder qi deficiency syndrome.Conclusion There are five common TCM syndromes of insomnia,and the characteristic symptoms of each TCM syndrome provide a reference source for the theoretical framework of syndrome diagnosis and efficacy evaluation scale and the establishment of item pool.

Objective:To compare the clinical effects of using xenogenic bone graft materials with or without collagen components for tooth micro crestal flap-alveolar ridge preservation (MCF-ARP) at molars with severe periodontitis.Methods:This study included patients who visited Department of Periodontology, Peking University School and Hospital of Stomatology from May to November 2023 due to severe periodontitis, requiring tooth extraction and planning for implant-retained prostheses. A total of 24 molars from 24 patients with severe periodontitis were assigned into two groups: the deproteinized bovine bone mineral (DBBM) group and the DBBM with collagen (DBBM-C) group. Twelve affected teeth from 12 patients were included in each group. Both groups underwent minimally invasive tooth extraction and MCF-ARP, with DBBM and DBBM-C implanted in the extraction socket, respectively. Cone beam CT (CBCT) was performed before and 6 months after the surgery for assessing changes of hard tissue. Intraoral scanning was performed before and at 2 weeks, 1 month, 3 months, and 6 months after the surgery for assessing soft tissue contour changes and patterns in both groups.Results:After 6 months of healing, the central bone height increased by (8.35±2.25) mm in the DBBM group and (7.70±2.36) mm in the DBBM-C group. The ridge width at 1 mm apically from the higher bone crest increased by 6.43 (-0.76,7.96) mm in the DBBM group and 6.01 (5.41,7.90) mm in the DBBM-C group. There was no statistically significant difference in the changes of bone height and width between the two groups (all P>0.05). In terms of soft tissue contour changes, although the buccal contour collapses were less in the DBBM-C group, the two groups showed no statistically significant difference (all P>0.05). Conclusions:Within the limitations of this study, it was demonstrated that the clinical effects of MCF-ARP using xenogenic bone graft materials with or without collagen components in molars with severe periodontitis were comparable.

Objective:To analyze the clinical characteristics and gene mutations of 4 patients with the male phenotype of 17α-hydroxylase /17, 20-lyase deficiency(17-OHD), in order to improve the recognition and appropriate management of atypical cases.Methods:A retrospective analysis was performed on the clinical features, biochemical findings, and gene mutations of 4 patients with the male phenotype of 17-OHD treated in our hospital between 2018 and 2023.Results:The social gender of all 4 patients with 17-OHD was male. None of the 4 patients had hypertension or hypokalemia, but all had micropenis and gynecomastia. Two patients had adrenal hyperplasia, while adrenal morphology was normal in the other two. One patient had decreased bone mass. There were typical changes in the steroid synthesis-related hormone spectrum: progesterone was significantly elevated in all 4 patients, 17-hydroxyprogesterone was not markedly abnormal, cortisol, dehydroepiandrosterone sulfate(DHEAS) and estradiol levels were low, and testosterone levels were also low.Conclusion:17-OHD is a rare type of congenital adrenal hyperplasia, with the male phenotype being even rarer. Early symptoms are often atypical, resulting in high rates of misdiagnosis and missed diagnosis. Patients without hypertension and hypokalemia are particularly prone to diagnostic confusion. Male patients with unexplained progesterone elevation, poor secondary sexual development, and gynecomastia should undergo timely steroid hormone profiling and genetic testing to avoid misdiagnosis and missed diagnosis.

Objective:To investigate the efficacy of radiotherapy in patients with advanced esophageal cancer receiving first-line chemoimmunotherapy.Methods:A retrospective analysis was conducted on the data of 137 patients with Stage Ⅳ esophageal squamous cell carcinoma (ESCC) treated at our hospital from January 2018 to May 2023. These patients were divided into two groups: a group treated with first-line chemoimmunotherapy combined with radiotherapy (chemoimmunotherapy + radiotherapy group, n = 43) and a group treated with only chemoimmunotherapy ( n = 94). Inverse probability of treatment weighting (IPTW) was applied to balance baseline characteristics between the groups. With overall survival (OS) and progression-free survival (PFS) as study endpoints, the survival data were analyzed using the Kaplan-Meier method, the log-rank test, and the Cox regression method. Results:Before calibration, the chemoimmunotherapy + radiotherapy group significantly outperformed the sole chemoimmunotherapy group in median PFS (13.6 months vs. 7.0 months; HR: 0.501, 95% CI: 0.309-0.811, P = 0.005). After calibration using the COX proportional-hazards model for age, gender, Eastern Cooperative Oncology Group (ECOG) performance status, smoking history, T/N/M stage, and tumor location, the chemoimmunotherapy + radiotherapy group still had significant advantages in PFS (14.7 months vs. 7.0 months; HR: 0.441, 95% CI: 0.261-0.745, P = 0.002). IPTW analysis further confirmed this trend (13.9 months vs. 7.0 months; HR: 0.492, 95% CI: 0.304-0.795, P < 0.001). Specifically, the median OS of the chemoimmunotherapy + radiotherapy group demonstrated significant improvement in all analyses: pre-calibration (29.5 months vs. 18.0 months; HR: 0.507, 95% CI: 0.297-0.867, P = 0.013), after calibration using the Cox model (27.5 months vs. 16.7 months; HR: 0.470, 95% CI: 0.266-0.830, P = 0.009), and after calibration using IPTW (29.5 months vs. 16.9 months; HR: 0.448, 95% CI: 0.262-0.764, P < 0.001). Conclusions:The combination of radiotherapy and first-line chemoimmunotherapy can significantly improve survival outcomes of patients with advanced ESCC, suggesting its potential as a standard treatment strategy.