Objective : To observe the brain tissue injury during hypoxia-ischemia, as well as the pathological changes and the expression of ferroptosis-related factors after the use of Shenfu injection(SFI), and to explore the protective effect of SFI on hypoxic-ischemic brain injury(HIBD) by inhibiting ferroptosis. Methods : An animal model of HIBD in SD rats was constructed and intervened with SFI. Pathologic changes in brain tissue were observed by HE staining methods. Nissen staining was used to observe neuron survival. Glutathione Peroxidase 4(GPX4) and Divalent Metal Transporter 1(DMT1) expression were detected in brain tissue by Western blot, immunohistochemistry and immunofluorescence. Reduced Glutathione(GSH), Lactate Dehydrogenase(LDH), Malondialdehyde(MDA), Superoxide Dismutase(SOD) and tissue iron content were determined with the kits. BV-2 microglial cell line(BV2) cells were culturedin vitroand divided into control group(Ctrl group), oxygen-glucose deprivation group(OGD group), iron ferroptosis-inducing group(Erastin group), iron ferroptosis-inhibiting group(Fer-1 group), Shenfu injection group(SFI group), and Erastin+Shenfu injection group(Erastin+SFI group). 2′,7′-Dichlorodihydrofluorescein diacetate(DCFH-DA) reactive oxygen species(ROS) fluorescent probe was used to detect the ROS release level; Immunofluorescence was used to observe intracellular GPX4, DMT1 expression. Results : Compared with the Sham group, rats in the HIBD group showed significant neuronal cell damage in brain tissue, decreased GPX4 expression(P<0.01), increased DMT1 expression(P<0.01), decreased GSH and SOD levels(P<0.01), and increased LDH, MDA and tissue iron levels(P<0.05,P<0.05,P<0.01). In contrast, after the intervention of SFI, GPX4 expression was elevated(P<0.01), DMT1 expression decreased(P<0.01), GSH and SOD levels were elevated(P<0.01), and LDH, MDA, and tissue iron levels decreased(P<0.05,P<0.05,P<0.01). The cells experiments showed that compared with the Ctrl group, the OGD group had a significantly higher ROS content and a decrease in the expression of GPX4 fluorescence intensity, and an increase in the fluorescence intensity of DMT1(P<0.01), compared with the OGD group, the ROS content was reduced in the SFI group, while the expression of GPX4 was elevated and the expression of DMT1 was reduced(P<0.01). Conclusion Hippocampal and cortical regions are severely damaged after HIBD in neonatal rats, and their brain tissues show decreased expression of GPX4 and increased expression of DMT1. The above suggests that ferroptosis is involved in HIBD brain injury in neonatal rats. In contrast, Shenfu injection has a protective effect on HIBD experimental animal model and BV2 cell injury model by reducing iron aggregation and ROS production.

Objective To investigate whether cisplatin induces tumor necrosis factor-α(TNF-α)secretion in human head and neck squamous cell carcinoma(HNSCC)cells to trigger RIP1/RIP3/MLKL-dependent necroptosis of the cells.Methods HNSCC cell lines HN4 and SCC4 treated with cisplatin(CDDP)or the combined treatment with CDDP and z-VAD-fmk(a caspase inhibitor)or Nec-1(a necroptosis inhibitor)for 24 h were examined for changes in cell viability using CCK8 assay and expressions of caspase-8 and necroptosis pathway proteins(RIP1/RIP3/MLKL)using Western blotting.The changes in migration of the cells were assessed with cell scratch assay,and the expressions of epithelial-mesenchymal transition(EMT)marker proteins N-cadherin,vimentin,and E-cadherin as well as the expressions of NF-κB(p65)and TNF-α were detected with Western blotting.Results The IC50 of cisplatin was 10 μg/mL in HN4 cells and 15 μg/mL in SCC4 cells.Cisplatin treatment significantly decreased the expressions of caspase-8,N-cadherin and vimentin and increased the expressions of E-cadherin,the necroptosis pathway proteins(RIP1/RIP3/MLKL),TNF-α,and NF-κB(p65),and these changes were obviously inhibited by treatment with Nec-1.Cisplatin stimulation also significantly lowered migration of the cells,and this inhibitory effect was strongly attenuated by Nec-1 treatment.Conclusion Cisplatin activates nuclear factor-κB signaling in HNSCCs to promote TNF-α autocrine and induce RIP1/RIP3/MLKL-dependent necroptosis,thus leading to inhibition of cell proliferation.

Twelve DEK-NUP214 fusion gene-positive patients with acute myeloid leukemia and on allo-HSCT treatment at the Hematology Hospital of the Chinese Academy of Medical Sciences from November 2016 to August 2022 were included in the study, and their clinical data were retrospectively analyzed. The patients comprised five men and seven women with a median age of 34 (16-52) years. At the time of diagnosis, all the patients were positive for the DEK-NUP214 fusion gene. Chromosome karyotyping analysis showed t (6;9) (p23;q34) translocation in 10 patients (two patients did not undergo chromosome karyotyping analysis), FLT3-ITD mutation was detected in 11 patients, and high expression of WT1 was observed in 11 patients. Nine patients had their primary disease in the first complete remission state before transplantation, one patient had no disease remission, and two patients were in a recurrent state. All patients received myeloablative pretreatment, five patients received sibling allogeneic hematopoietic stem cell transplantation, and seven patients received haploid hematopoietic stem cell transplantation. The median number of mononuclear cells in the transplant was 10.87 (7.09-17.89) ×10 8/kg, and the number of CD34 + cells was 3.29 (2.53-6.10) ×10 6/kg. All patients achieved blood reconstruction, with a median time of 14 (10-20) days for neutrophil implantation and 15 (9-27) days for platelet implantation. The 1 year transplant-related mortality rate after transplantation was 21.2%. The cumulative recurrence rates 1 and 3 years after transplantation were 25.0% and 50.0%, respectively. The leukemia free survival rates were (65.6±14.0) % and (65.6±14.0) %, respectively. The overall survival rates were (72.2±13.8) % and (72.2±13.8) %, respectively.

Objective To investigate whether cisplatin induces tumor necrosis factor-α(TNF-α)secretion in human head and neck squamous cell carcinoma(HNSCC)cells to trigger RIP1/RIP3/MLKL-dependent necroptosis of the cells.Methods HNSCC cell lines HN4 and SCC4 treated with cisplatin(CDDP)or the combined treatment with CDDP and z-VAD-fmk(a caspase inhibitor)or Nec-1(a necroptosis inhibitor)for 24 h were examined for changes in cell viability using CCK8 assay and expressions of caspase-8 and necroptosis pathway proteins(RIP1/RIP3/MLKL)using Western blotting.The changes in migration of the cells were assessed with cell scratch assay,and the expressions of epithelial-mesenchymal transition(EMT)marker proteins N-cadherin,vimentin,and E-cadherin as well as the expressions of NF-κB(p65)and TNF-α were detected with Western blotting.Results The IC50 of cisplatin was 10 μg/mL in HN4 cells and 15 μg/mL in SCC4 cells.Cisplatin treatment significantly decreased the expressions of caspase-8,N-cadherin and vimentin and increased the expressions of E-cadherin,the necroptosis pathway proteins(RIP1/RIP3/MLKL),TNF-α,and NF-κB(p65),and these changes were obviously inhibited by treatment with Nec-1.Cisplatin stimulation also significantly lowered migration of the cells,and this inhibitory effect was strongly attenuated by Nec-1 treatment.Conclusion Cisplatin activates nuclear factor-κB signaling in HNSCCs to promote TNF-α autocrine and induce RIP1/RIP3/MLKL-dependent necroptosis,thus leading to inhibition of cell proliferation.

Objective To investigate the efficacy of different courses of saccharomyces boulardii powder combined with triple therapy in eradicating helicobacter pylori(Hp)in children.Methods A total of 135 children with Hp-related gastritis who received initial treatment in the Department of Pediatrics,Affiliated Hospital of Xuzhou Medical University from October 2021 to June 2022 were selected and divided into three groups according to random number table method:group A,group B and group C,with 45 cases in each group.Group A:triple therapy(omeprazole + clarithromycin + amoxicillin)for 14 days;group B and group C:on the basis of triple therapy,saccharomyces boulardii powder was added from the first day of treatment for 2 weeks and 4 weeks,respectively.The adverse reactions during treatment were recorded,the levels of serum pepsinogen Ⅰ(PGⅠ)and pepsinogen Ⅱ(PGⅡ),clinical efficacy and Hp eradica-tion rate were observed 4 weeks after the end of treatment.Results The levels of serum PGⅠ and PGⅡ in the three groups after treat-ment were significantly lower than those before treatment(P<0.05),the levels of serum PGⅠ and PGⅡ in group B and group C after treatment were lower than those in group A(P<0.05),and the levels of serum PGⅠ and PGⅡ in group B were lower than those in group C after treatment(P<0.05).The clinical effective rates of group B(93.0%)and group C(90.4%)were higher than those of group A(62.5%)(P<0.05).The clinical effective rate of group B was higher than that of group C(P>0.05).The Hp eradication rate of group B and group C was significantly higher than that of group A(P<0.05),and the Hp eradication rate of group B was higher than that of group C(P>0.05).The incidence of diarrhea and loss of appetite in group B and group C were lower than those in group A(P<0.05);the incidence of diarrhea and loss of appetite in group B was lower than that in group C(P<0.05).There was no signifi-cant difference in the incidence of abdominal pain,nausea and vomiting among the three groups(P>0.05).Conclusion Saccharomy-ces boulardii decoction combined with triple therapy for 2 or 4 weeks can effectively regulate PG level,improve the eradication rate of Hp,and reduce the incidence of adverse reactions.It is suggested that 2 weeks is the best course of treatment.

Objective:To investigate the effect of helicobacter pylori (HP) infection and eradication treatment on small intestinal bacterial overgrowth (SIBO) in children.Methods:A prospective case-control study was conducted to select 68 children with symptoms of abdominal distension, abdominal pain, diarrhea and suspected digestive system diseases admitted to the Affiliated Hospital of Xuzhou Medical University from June 2021 to June 2022. They were divided into HP negative group and HP positive group according to HP infection. HP positive group received triple standardized HP eradication treatment, 14 days as a course of treatment. The baseline SIBO positive rate and gastrointestinal symptom rating scale (GSRS) score of the two groups were compared. The HP positive group was followed up for 4 and 12 weeks after drug withdrawal for quantitative assessment of gastrointestinal symptoms and LHBT. The SIBO positive rate, GSRS score of the two groups and the change of SIBO positive rate and GSRS score of the HP positive group before and after treatment were compared. The measurement data with normal distribution were expressed, and independent sample t-test was used for comparison between the two groups. M( Q1, Q3) was used to represent the measurement data of non normal distribution, and Mann Whitney U test was used to compare the two groups; Friedman test was used for comparison between multiple time points, and Nemenyi test was used for pairwise comparison. Four grid table or paired χ 2 test was used to compare the counting data between groups. Results:The positive rate of SIBO in HP negative group was lower than that in HP positive group (36.1% (13/36) vs 62.5% (20/32)), the difference was statistically significant (χ 2=4.72, P=0.030). Four weeks after drug withdrawal, the SIBO positive rate in HP positive group was higher than that before treatment (87.5% (28/32) vs 62.5% (20/32)), and 12 weeks after drug withdrawal was lower than that before treatment (21.9% (7/32) vs 62.5% (20/32)), with statistically significant differences (χ 2=8.00, P=0.008; χ 2=13.00, P<0.001). There was no statistically significant difference in GSRS score between HP negative group and HP positive group ( P=0.098). The clinical symptoms of 32 children in HP positive group were improved 4 and 12 weeks after HP eradication was stopped. GSRS scores were lower than those before treatment (8.0 (6.0, 12.8), 7.0 (5.0, 9.0) points vs 15.0 (12.0, 19.0) points) , and the differences were statistically significant ( Z values were -3.91, -4.68, respectively; all P<0.001). Conclusions:HP infection can increase the positive rate of SIBO in children with suspected digestive system diseases. The standardized triple HP eradication therapy may further aggravate the overgrowth of intestinal bacteria while treating HP infection, but this effect can be eliminated after 12 weeks of treatment.

Objective:To explore the expression of the nucleotide-binding oligomerization domain-like receptor containing pyrin domain protein 6 (NLR family,pyrin domain containing 6,NLRP6) in the gastric tissue and gastric juice of children with chronic non-atrophic gastritis (CNG), and to analyze the influence of Helicobacter pylori (Hp) infection on the expression of NLRP6.Methods:A case-control study was conducted to select 120 CNG patients in pediatrics of Department of Pediatrics, The Affiliated Hospital of Xuzhou Medical University from October 2020 to July 2021. According to pathological diagnosis, endoscopic gastric mucosal damage and Hp infection, they were divided into 4 groups: mild CNG group Hp negative, Moderate to severe CNG group Hp negative, Mild CNG group Hp positive, Moderate to severe CNG group Hp positive. The enzyme-linked immunosorbent assay (ELISA) was used to detect the expression level of NLRP6 in the four groups of gastric tissue and gastric juice, and Western blot was used to detect the expression of NLRP6 in the gastric tissue of the 4 groups, and the significance of expression in CNG of children is analyzed. Independent sample t-test was used to compare the mean between the two groups. One way ANOVA was used to compare the mean of multiple groups of samples, and LSD t-test was used for pairwise comparison. Comparison between count data groups χ 2 inspection. Results:The positive rate of Hp in the moderate to severe chronic non-atrophic gastritis group was 62.96% (34/54) higher than that in the mild chronic non-atrophic gastritis group 37.04% (20/54), and the difference was statistically significant (χ 2=18.32, P<0.001). Under the same Hp conditions, the expression of NLRP6 in the mild chronic non-atrophic gastritis group (Hp negative mild CNG: gastric tissue (653.73±37.71) ng/L, gastric juice (471.75±38.47) ng/L; Hp positive mild CNG: Gastric tissue (616.69±43.33) ng/L, gastric juice (445.29±36.39) ng/L was higher than the moderate to severe chronic non-atrophic gastritis group (Hp negative moderate to severe CNG: gastric tissue (623.82±52.99) ng/L, gastric juice (446.48±47.49) ng/L; Hp positive Moderate to severe CNG: gastric tissue (580.43±62.75) ng/L, gastric juice (406.88±51.85) ng/L, the difference is statistically significant (under Hp negative, mild compared with moderate to severe CNG: gastric tissue P=0.035; gastric juice P=0.046; Under Hp positive, mild compared with moderate to severe CNG: gastric tissue P=0.010;gastric juice P=0.002); in the same degree of gastric mucosal injury, NLRP6 expression in Hp-negative group (Hp-negative mild CNG: gastric tissue (653.73±37.71) ng/L, gastric juice (471.75±38.47) ng/L; Hp negative moderate to severe CNG: gastric tissue (623.82±52.99) ng/L, gastric juice (446.48±47.49) ng/L higher than the positive group (Hp positive mild CNG: gastric tissue (616.69±43.33) ng/L, gastric juice (445.29±36.39) ng/L; Hp positive moderate to severe CNG: gastric tissue (580.43±62.75) ng/L, gastric juice (406.88±51.85) ng/L, the difference is statistically significant (under mild CNG, Hp negative is compared with positive: Gastric tissue P=0.005; gastric juice P=0.023; under moderate to severe CNG, negative versus positive: gastric tissue P=0.004; gastric juice P=0.003). Conclusion:Under the same Hp conditions, the more severe the gastric mucosal damage, the lower the NLRP6; under the same degree of mucosal damage, the expression level of NLRP6 in the Hp-negative group was significantly higher than that of the Hp-positive group. It is suggested that NLRP6 plays a role in inhibiting inflammation in chronic gastritis, maintaining the integrity of epithelial cells, and Hp can inhibit the expression of NLRP6.

China ; Humans ; Lenalidomide/therapeutic use* ; Multiple Myeloma/drug therapy* ; Thalidomide ; Therapeutic Equivalency

Objective: To evaluate the outcomes of myelodysplastic syndromes (MDS) patients who received HLA-matched sibling donor allogeneic peripheral blood stem cell transplantation (MSD-PBSCT) . Methods: The clinical data of 138 MDS patients received MSD-PBSCT from Sep. 2005 to Dec. 2017 were retrospectively analyzed, and the overall survival (OS) rate, disease-free survival (DFS) rate, relapse rate (RR) , non-relapse mortality (NRM) rate and the related risk factors were explored. Results: ①After a median follow-up of 1 050 (range 4 to 4 988) days, the 3-year OS and DFS rates were (66.6±4.1) % and (63.3±4.1) %, respectively. The 3-year cumulative incidence of RR and NRM rates were (13.9±0.1) % and (22.2±0.1) %, respectively. ②Univariate analysis showed that patients with grade Ⅲ-Ⅳ acute graft-versus-host disease (aGVHD) or hematopoietic cell transplantation comorbidity index (HCT-CI) ≥2 points or patients in very high-risk group of the Revised International Prognostic Scoring System (IPSS-R) had significantly decreased OS[ (42.9±13.2) %vs (72.9±4.2) %, χ²=8.620, P=0.003; (53.3±7.6) %vs (72.6±4.7) %, χ²=6.681, P=0.010; (53.8±6.8) %vs (76.6±6.2) %vs (73.3±7.7) %, χ²=6.337, P=0.042]. For MDS patients with excess blasts-2 (MDS-EB2) and acute myeloid leukemia patients derived from MDS (MDS-AML) , pre-transplant chemotherapy or hypomethylating agents (HMA) therapy could not improve the OS rate[ (60.4±7.8) %vs (59.2±9.6) %, χ²=0.042, P=0.838]. ③Multivariate analysis indicated that the HCT-CI was an independent risk factor for OS and DFS (P=0.012, HR=2.108, 95%CI 1.174-3.785; P=0.008, HR=2.128, 95%CI 1.219-3.712) . Conclusions HCT-CI was better than the IPSS-R in predicting the outcomes after transplantation. The occurrence of grade Ⅲ-Ⅳ aGVHD is a poor prognostic factor for OS. For patients of MDS-EB2 and MDS-AML, immediate transplantation was recommended instead of receiving pre-transplant chemotherapy or HMA therapy.

Objective: To evaluate the outcomes of human leukocyte antigen (HLA) matched unrelated donor hematopoietic stem cell transplantation (MUD-HSCT) for adult acute myeloid leukemia (AML) in a single center. Methods: Consecutive adult AML who received MUD-HSCT in our center from January 2008 to April 2017 were studied retrospectively, comparing with patients undergoing matched sibling donor (MSD) -HSCT in the same period. The rates of overall survival (OS) , disease free survival (DFS) , relapse, non-relapse mortality (NRM) , engraftment, acute and chronic graft-versus-host disease (aGVHD and cGVHD) were analyzed. Results: A total of 247 consecutive cases were enrolled, including 46 patients with MUD-HSCT and 201 with MSD-HSCT. All the patients experienced neutrophil engraftment except for one patient who died early in the MSD group, but the median day of engraftment was longer in the MUD group (15.0 vs 14.0, P=0.017) . The accumulative engraftment rate of platelet was comparable between the two groups (93.5%vs 98.0%, P=0.128) . The accumulative incidences of aGVHD (50.0%vs 46.3%, P=0.421) and cGVHD (37.8%vs 43.0%, P=0.581) were not statistically different between the two groups. Compared with the MSD group, the accumulative NRM rate at+36 months after transplantation was significantly higher in the MUD group (22.0%vs 10.4%, P=0.049) , while the relapse rate was not statistical difference (20.5 vs 28.3%, P=0.189) . Both the 3-year OS (61.6%vs 63.3%, P=0.867) and DFS (57.5%vs 61.6%, P=0.760) were comparable between the two groups. Four independent risk factors were confirmed by the multivariate analysis: patient age ≥45 years old, CR2 or NR before transplantation, a history of extramedullary infiltration and the occurrence of grade Ⅲ-Ⅳ aGVHD. No statistical differences were demonstrated in the survival rate between MUD-and MSD-HSCT in different subgroups. Conclusions The outcomes, such as GVHD, relapse, OS and DFS, were comparable between MUD-and MSD-HSCT for adult AML, but higher incidence of NRM and longer time to neutrophil engraftment in the MUD group. MUD-HSCT is practical and feasible for adult AML who are lack of MSD.