Objective To investigate the effect of bFGF on the grafting ability (grafing efficiency)of CD34 + cord blood cells seeded in 5 FU injured human bone marrow stromal cells.Methods Using combined methods of bone marrow stromal cell culture system,light microscopy and electron microscopy,flow cytometry,we investigated the toxic effects of 5 FU on human bone marrow stromal cells (hBMSC) and the benefit of bFGF on dynamics of hBMSC as well as the ability of the cord blood stem cell to bind hBMSC.Results After 3 hours incubation with 5 FU,significant changes in hBMSC cytosolic and nuclear morphology were observed.Nuclear splitting and abnormal shapes of the pseudopodia were found under the electron microscope.Flow cytometry revealed that apoptosis appeared before G 0/G 1,and that cells in G 0/G 1and G 2/M phases were remarkably lower,while the cells in S phase were remarkably higher than the controls.Two hours after adding the cord blood hematopoietic progenitor cells,the group treated with bFGF(injury with intervention of bFGF group,IBG)showed higher proprtion of the adherence layer compared to the group without adding bFGF(injury group,IG)(29% versus 12%) Conclusion 5 FU causes DNA damage in hBMSC.It also impairs the binding ability of cord blood stem cell to hBMSC.bFGF is able to repair hBMSC damage and improve the binding of CD34 + cord blood hematopoietic progenitor cells onto hBMSC.

Objective To elucidate the role of human bone marrow stromal cells (hBMSC) in combination with exogenous cytokines such as stem cell factor(SCF) and FLT 3 ligand (FL) in supporting the in vitro expansion of CD 34 + cells purified from cord blood.Methods CD 34 + cells were isolated from umbilical cord blood by using a high gradient magnetic cell sorting system(MACS),expanded in combination with SCF,IL3 (interleukin 3),IL 6 (interleukin 6),FL,EPO (erythropoietin),and were planted onto the pre established irradiated (20Gy) stroma layer(hBMSC plus combinations of cytokines) respectively.On day 10,total cells were counted, and hematopoietic progenitor cells were assessed by semisolid culture assay,CD 34 + cells were quantitated by FACS.Results After separation by MACS,the frequency of CD 34 + cells was 92%?0.04%.On day 2,almost all the inoculated cells adhered to the stroma layer,with only a small number in the supernatant.Then,cells in the supernatant increased gradually, but the percentage of CD 34 + cells decreased.Compared with control, expansion fold of CD 34 + cells, CFU GM, BFU E were significantly higher in hBMSC group (P 0.5).Conclusion ① CD 34 + cells from cord blood formed foci adherent to the monolayer and colony forming cells remained in the culture of 10days,indicating that hBMSC can support hematopoiesis in vitro;②using human bone marrow stromal cells in cooperation with exogenous cytokines may be a feasible way to expand hematopoietic stem/progenitor cells.

Objective To evaluate the curative effect of traditional Chinese medicine fumigation combined with conventional therapy for the diabetic foot. Methods A total of 90 patients with type 2 diabetes who were treated in the hospital from March 2015 to August 2017 were enrolled in this study. On the basis of conventional treatment, the treatment group received the traditional Chinese medicine fumigation, while the control group received warm water fumigation. The total limb popliteal artery, femoral artery, posterior tibial artery and dorsalis pedis artery and a score of symptoms and signs were compared between two groups after 4 weeks. Results The total effective rate of the treatment group was 73.3% (33/45), while the control group was 53.3% (24/45), and the difference was statistically significant (χ2=3.876, P=0.049). After treatment, total limb femoral artery(58.38 ± 12.53 cm/s vs.51.94 ± 11.62 cm/s,t=2.528),popliteal artery(43.87 ± 9.46 cm/s vs.39.87 ± 8.28 cm/s,t=2.134),posterior tibial artery(35.72 ± 8.29 cm/s vs.30.89 ± 8.51 cm/s,t=2.725),dorsalis pedis artery (26.82 ± 8.10 cm/s vs.22.87 ± 7.39 cm/s,t=2.413)of the treatment group were significantly higher than those of the control group(P<0.05).The main symptom score(2.0 ± 1.5 vs.4.1 ± 2.1,t=5.419),sub syndrome score(3.1 ± 1.8 vs.5.1 ± 2.3,t=4.692),tongue and pulse condition score(1.8 ± 0.8 vs.2.4 ± 0.6,t=4.277)of the treatment group were significantly lower than those of the control group (P<0.01). Conclusions The traditional Chinese medicine fumigation combined with conventional therapy could improve the lower extremity blood flow status in diabetic foot patients and clinical symptoms.

Objective:To summarize the clinical and gene mutation characteristics of a child with Cowden syndrome and review the literature.Methods:The clinical data of a child with Cowden syndrome treated in the First Affiliated Hospital of Xinxiang Medical University in June 2020 were analyzed retrospectively.Taking " Cowden syndrome" , " PTEN gene" , " hamartoma polyps" , "child" , " Cowden syndrome and child" and " PTEN and child" as key words, literature was retrieved from Chinese databases (China National Knowledge Internet database and Wanfang database) and the PubMed database from the establishment of the database to March 2021. Results:A 13-year-old male had intermittent abdominal pain and abdominal distension for 5 months.Electron microscopic gastroenteroscopy showed multiple polyps, and focal lymphocyte aggregation and infiltration were found in tissue biopsy.Whole exon sequencing revealed a hemizygous mutation of c. 475 (exon5) A>T in PTEN gene, which led to the transformation of the 159 th amino acid from arginine to tryptophan.The prediction results of the tertiary structure of the protein indicated that the variation might affect the spatial structure of the protein and damage the protein function.According to the clinical characteristics, Cowden syndrome was diagnosed.The pedigree confirmed that the variation was inherited from the mother, who had a similar phenotype.No qualified Chinese report was retrieved.Among 41 English studies on PTEN gene mutation in children, there were only two reports related to pediatric Cowden syndrome.The hemizygous mutation of PTEN gene was not reported. Conclusions:The missense mutation of c. 475 (exon5) A>T in PTEN gene in this study is a novel cause of Cowden syndrome, and the case is the first case report in China.

AIM:This study investigates the apoptotic and autophagic effects of atractylodin on lung cancer cells,elucidating the underlying molecular mechanisms.METHODS:Non-small cell lung cancer(NSCLC)A549 and H460 cells,in addition to non-cancerous HBE cells,were cultured in vitro.The effects of atractylodin at various concen-trations on cell viability were assessed using CCK-8 assay.Apoptotic effects were evaluated through Hoechst staining and flow cytometry,while Western blot analysis was performed to detect changes in protein expressions associated with apopto-sis and autophagy,including P62,beclin-1,microtubule-associated protein 1 light chain 3(LC3),Kelch-like epichloro-hydrin(ECH)-associated protein-1(Keap-1),nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HO-1),and NAD(P)H:quinone oxidoreductase 1(NQO1).Autophagic flux was further analyzed using acridine orange(AO)stain-ing,and immunofluorescence for LC3 and Nrf2.Additionally,autophagy inhibition experiments were conducted using chloroquine(CQ),followed by analyses of autophagy and apoptosis.Reactive oxygen species(ROS)levels were quanti-fied using DCFH-DA.RESULTS:Treatment with atractylodin significantly reduced the viability of A549 and H460 lung cancer cells,promoting apoptosis and inducing autophagy.This was evidenced by an increase in acidic autophagic vesi-cles,upregulation of LC3 and beclin-1,and downregulation of P62.Inhibition of autophagy by chloroquine reversed atrac-tylodin-induced apoptosis.Moreover,atractylodin heightened ROS production,inhibited Keap-1,and stimulated the ex-pression of Nrf2,HO-1 and NQO1.CONCLUSION:Atractylodin effectively inhibits the proliferation of lung cancer cells by inducing apoptosis and autophagy.These effects are mediated through the modulation of the ROS/Nrf2/HO-1 sig-naling pathway,underscoring its potential as a therapeutic agent in lung cancer treatment.

Objective:To observe the efficacy and adverse reactions of adrenocorticotropic hormone (ACTH) in the treatment of recurrent frequently relapsing nephrotic syndrome (FRNS), and explore the feasibility of treatment of ACTH in children.Methods:From November 2017 to June 2018, in the First Affiliated Hospital of Xinxiang Medical University of a total of 32 cases of FRNS ACTH therapy were all the role of ACTH consecutive 3-8 courses of treatment (when the dosage of prednisone was less than or equal to 0.5 mg/kg, 0.4 U/kg ACTH should be used every day.After 3 days of continuous application, the dosage of Prednisone should be reduced to 1.25-5.00 mg.ACTH was used for 3 days as a course of treatment, and continued to use ACTH for 2 courses until corticosteroid stopped). The number of recurrence, Prednisone maintenance dose, immunosuppressive use, serum cortisol and other relevant experimental indicators and adverse reactions were recorded during the follow-up period after ACTH treatment, and were followed up for 8-15 months.Results:Among the 32 children, the onset age (53.47±25.81) months, the course of disease (35.25±23.87) months, 22 patients (68.75%) had no recurrence after corticosteroid withdrawal, 7 patients (21.87%) had recurrence after corticosteroid withdrawal, and 3 patients (9.37%) had recurrence during corticosteroid withdra-wal.Compared with before ACTH treatment, Prednisone was significantly decreased[(0.08±0.14) mg/(kg·d) vs. (0.23±0.23) mg/(kg·d)], and the difference was statistically significant ( t=3.661, P<0.05), the number of immunosuppressant cases decreased significantly[42.38%(12/32 cases) vs. 58.62%(17/32 cases), χ2= 14.500, P<0.05]. Serum cortisol was measured at 8 Am increased significantly[(11.78±4.64) μg/dL vs. (4.42±3.13) μg/dL, t=7.340, P<0.05]. The results were stable during follow-up, with 2 patients presenting with systemic urticaria once and 1 patient presenting with headache after infusion. Conclusions:ACTH therapy is safe and effective in hormone therapy for child with FRNS, and less adverse reactions.

A case of primary amoebic meningoencephalitis (PAM) treated in the First Affiliated Hospital of Xinxiang Medical University in March 6, 2022 was reported.The proband was a 14-year-old boy, who was admitted to the hospital because of " fever, headache and vomiting for 2 days" . Metagenomic next-generation sequencing (mNGS) was consequently conducted to examine the pathogens in cerebrospinal fluid.Naegleria infection was detected, so the child was diagnosed with PAM.The disease developed rapidly, and the patient died 29 hours after admission.In the paper, a total of 13 studies were reviewed, and 15 children with PAM were reported.Of the reported cases, only 1 case survived, 14 cases died.PAM had a low incidence, a dangerous condition, and high mortality.Most cases were diagnosed by autopsy or pathogen diagnosis in cerebrospinal fluid.This case is confirmed by mNGS of pathogens, and it is rarely reported at home and abroad.

Objective:To investigate the effects of radiation dose to the host immune system during radiotherapy on disease progression and survival in patients with peripheral early-stage non-small cell lung cancer (ES-NSCLC) receiving stereotactic body radiation therapy (SBRT).Methods:Clinical data of pathologically confirmed ES-NSCLC patients who were treated with SBRT at Tianjin Medical University Cancer Institute and Hospital between January 2007 and December 2020 were retrospectively analyzed. The prognostic significance of the estimated dose of radiation to immune cells (EDRIC) in ES-NSCLC patients undergoing SBRT was cited and validated. EDRIC was calculated using the model developed by Kong et al. and improved by Ladbury et al. Kaplan-Meier method and Cox proportional hazards regression were adopted to estimate cancer-specific survival (CSS), progression-free survival (PFS), local progression-free survival (LPFS), and distant metastasis-free survival (DMFS). Pearson's correlation was used to assess the correlation between variables. Results:The median prescription dose/fraction was 60 Gy/5 fractions (range: 48-60 Gy in 3-10 fractions). The median follow-up time was 52.17 (1.17-154.77) months. The median gross tumor volume (GTV) and EDRIC were 10.98 (0.91-120.34) cm 3 and 2.064 (0.426-6.015) Gy, respectively. Person's correlation analysis showed that GTV was positively correlated with EDRIC ( r=0.712, P<0.001). In multivariate analysis, EDRIC was an important prognostic variable of CSS and DMFS. Higher EDRIC was significantly associated with worse CSS ( HR=1.763, P=0.004) and DMFS ( HR=1.902, P=0.004). Compared to patients with EDRIC ≤ 1.56 Gy, those with EDRIC > 2.64 Gy and EDRIC between <2.06-2.64 Gy exhibited significantly lower CSS ( P<0.001, P=0.049). There were significant differences in DMFS among the groups divided by quartiles of EDRIC (compared to EDRIC ≤1.56 Gy, the P values were <0.001, 0.004, and 0.022 respectively). Conclusions:EDRIC is an important predictor of CSS and DMFS in ES-NSCLC patients treated with SBRT, suggesting that radiation dose to the immune system is a critical determinant of treatment outcomes. EDRIC can be used to quantify the effects of radiation therapy on the host immune system.