Virtual Screening and Testing of GSK-3 Inhibitors Using Human SH-SY5Y Cells Expressing Tau Folding Reporter and Mouse Hippocampal Primary Culture under Tau Cytotoxicity

Glycogen synthase kinase-3β (GSK-3β) is an important serine/threonine kinase that implicates in multiple cellular processes and links with the neurodegenerative diseases including Alzheimer’s disease (AD). In this study, structure-based virtual screening was performed to search database for compounds targeting GSK-3β from Enamine’s screening collection. Of the top-ranked compounds, 7 primary hits underwent a luminescent kinase assay and a cell assay using human neuroblastoma SH-SY5Y cells expressing Tau repeat domain (TauRD) with pro-aggregant mutation ΔK280. In the kinase assay for these 7 compounds, residual GSK-3β activities ranged from 36.1% to 90.0% were detected at the IC50 of SB-216763. In the cell assay, only compounds VB-030 and VB-037 reduced Tau aggregation in SH-SY5Y cells expressing ΔK280 TauRD-DsRed folding reporter. In SH-SY5Y cells expressing ΔK280 TauRD, neither VB-030 nor VB-037 increased expression of GSK-3α Ser21 or GSK-3β Ser9. Among extracellular signal-regulated kinase (ERK), AKT serine/threonine kinase 1 (AKT), mitogen-activated protein kinase 14 (P38) and mitogenactivated protein kinase 8 (JNK) which modulate Tau phosphorylation, VB-037 attenuated active phosphorylation of P38 Thr180/ Tyr182, whereas VB-030 had no effect on the phosphorylation status of ERK, AKT, P38 or JNK. However, both VB-030 and VB-037 reduced endogenous Tau phosphorylation at Ser202, Thr231, Ser396 and Ser404 in neuronally differentiated SH-SY5Y expressing ΔK280 TauRD. In addition, VB-030 and VB-037 further improved neuronal survival and/or neurite length and branch in mouse hippocampal primary culture under Tau cytotoxicity. Overall, through inhibiting GSK-3β kinase activity and/or p-P38 (Thr180/Tyr182), both compounds may serve as promising candidates to reduce Tau aggregation/cytotoxicity for AD treatment.

Mechanism of curcumin inhibiting choroidal neovascularization in brown Norway rat / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To investigate the mechanism of curcumin inhibiting the choroidal neovascularization(CNV)of brown Norway(BN)rats.METHODS: CNV model of 36 BN rats was established through laser photocoagulation induction, and they were divided into 6 groups with 6 rats in each group. Normal group was fed normally with no intervention, while 532nm laser photocoagulation was used to establish a experimental CNV model in BN rats. Rats after modeling were respectively intervened for 14d and divided into model group, ranibizumab group, curcumin low [100mg/(kg·d)], medium [200mg/(kg·d)], and high [400mg/(kg·d)] dose group. The model group was given intragastric administration of saline for 14d, ranibizumab(10mg/mL, 0.2mL/dose)was injected at 2d after photocoagulation with 5μL once for rats in ranibizumab group, and different concentrations of curcumin were intragastrically administrated to the rats in low, medium and high groups for 14d. Fundus photography, fundus fluorescein angiography(FFA)and indocyanine green angiography(ICGA)examination were performed at 14d after photocoagulation. Ocular histopathological specimens of rats with CNV were made, and the central thickness of CNV were observed by HE staining. Ocular histopathological specimens were made, and the expressions of AKT/p-AKT/HIF-1α/VEGF signaling pathway-related proteins were observed by immunohistochemistry. The mRNA relative expressions of AKT/HIF-1α/VEGF factor in CNV tissues were detected by RT-qPCR, and the protein expressions of AKT/p-AKT/HIF-1α/VEGF factor in CNV tissues were detected by Western-blot.RESULTS: CNV generation rates in the model group, the ranibizumab group, and the low, medium and high-dose curcumin groups were 78.18%, 73.21%, 77.19%, 75.86%, 74.55%, respectively, which were higher than 70%. The average absorbance were 182.12±6.59, 119.22±8.03, 166.45±8.33, 164.34±5.69, 149.22±6.45, respectively; the ranibizumab group was significantly lower than the model group(P<0.05); the low-dose, medium-dose and high-dose groups were significantly higher than the ranibizumab group(P<0.05), and the curcumin high-dose group was significantly lower than the model group(P<0.05). HE staining showed that the retinal tissue structure of BN rats in normal group was clear and neatly arranged. The central thickness of CNV in the ranibizumab group was significantly reduced at 14d after photocoagulation compared with the model group(P<0.05); While the curcumin high-dose group was significantly reduced compared with the model group(P<0.05), but increased when compared with ranibizumab group(P<0.05). Immunohistochemistry results showed that AKT, p-AKT, HIF-1α, and VEGF factors were negatively expressed in the retinal tissue structure of BN rats in the normal group, and no brown-yellow reactants were found. The expression of AKT, p-AKT, HIF-1α, and VEGF factors in the model group were higher than that in the normal group at 14d after photocoagulation(P<0.05); the ranibizumab group was lower than the model group(P<0.05). While the expression of the curcumin high-dose group was significantly decreased compared with the model group(P<0.05), but significantly increased when compared with ranibizumab group(P<0.05). The mRNA results showed that the relative expression levels of AKT, HIF-1α and VEGF mRNA in the model group at 14d after photocoagulation were higher than those of the normal group(P<0.05); the ranibizumab group was lower than the model group(P<0.05). While curcumin high-dose group was significantly decreased compared with the model group(P<0.05), but significantly increased when compared with ranibizumab group(P<0.05). Western-blot results showed that there was no significant difference in the relative expression of AKT protein among each experimental groups at 14d after photocoagulation. The relative expression of p-AKT protein in the model group was significantly higher than that in the normal group(P<0.05); the ranibizumab group was significantly lower than the model group(P<0.05); the curcumin high-dose group was significantly lower than the model group(P<0.05). The relative expression levels of HIF-1α protein were significantly higher in the model group than in the normal group(P<0.05), and the ranibizumab group was lower than in the model group(P<0.05). The relative expression levels of HIF-1α protein was lower in the curcumin high-dose group than in the model group(P<0.05)but higher than ranibizumab group(P<0.05). The relative expression level of VEGF protein was significantly lower in the curcumin medium/high-dose group than in the model group(P<0.05).CONCLUSION: Curcumin at 400mg/(kg·d)has an inhibitory effect on CNV in BN rats. The mechanism may be closely related to inhibiting the activation of AKT/p-AKT/HIF-1α/VEGF signaling pathways.

Current situation of treatment and disposal of NORM solid waste in China / 中国辐射卫生

@#China has a huge cumulative storage of NORM solid waste, which urgently needs to be treated and disposed of in a scientific and standard way. Through analyzing the current status and technologies of treatment and disposal of NORM solid waste and relevant environmental radiation protection regulations and standards, we put forward suggestions on the improvement of regulations and standards and the applicability of treatment and disposal technologies, to provide a reference for the green and sustainable development of mineral resources in China.

Correlation between platelet distribution width and anticoagulant stability of warfarin in patients with persistent atrial fibrillation / 中国医师杂志

Objective:To explore the correlation between platelet distribution width (PDW) and the stability of warfarin anticoagulant therapy in patients with persistent atrial fibrillation.Methods:138 patients with persistent atrial fibrillation treated with warfarin in Jiujiang First People′s Hospital from January 2018 to December 2019 were selected. They were divided into groups according to whether PDW increased (PDW decreased group, normal group, PDW increased group) and subgroups stratification was performed. After stratification, the relationship between PDW and the stability of warfarin anticoagulation treatment [expressed as the percentage of time of International normalized ratio(INR) within the treatment target range (TTR)] was analyzed. At the same time, the predictive value of PDW for the stability of warfarin anticoagulation treatment was analyzed.Results:There were significant difference in PDW and TTR among the PDW decreased group, normal group, PDW increased group ( F=30.322, 10.745, all P<0.01). The PDW distribution of patients with different anticoagulation quality was significantly different (χ 2=9.532, P<0.05). Receiver operating characteristic (ROC) curve showed that the area under curve (AUC) of PDW in predicting warfarin anticoagulant stability was 0.621(95% CI: 0.524-0.737). There was significant difference in PDW and TTR among the PDW<14%, 14%-<16%, 16-<18% and ≥18% groups( F=18.075, 11.638, all P<0.01). There was no significant difference in PDW and TTR among the three subgroups of PDW<14%, 14%-<16% and 16-<18% ( P=0.843, P=0.401). There were significant difference in PDW and TTR between the two subgroup of PDW 16-<18%、≥18% ( t=4.154, 6.712, all P<0.01). Conclusions:PDW is correlated with the standard rate of warfarin anticoagulant stability, and can be used to predict the standard rate of warfarin anticoagulant stability.

Efficacy of auricular acupoint bean-embedding therapy in the treatment of residual dizziness after successful reduction of benign paroxysmal positional vertigo and its effect on skin sympathetic response / 中国基层医药

Objective:To investigate the efficacy of auricular acupoint bean-embedding therapy in the treatment of residual dizziness after successful reduction of benign paroxysmal positional vertigo (BPPV) and its effect on skin sympathetic response.Methods:A total of 110 patients with residual dizziness after successful reduction of BPPV who were admitted to Wenzhou Hospital of Traditional Chinese Medicine from January 2019 to May 2021 were included in this study. They were randomly divided into control and study groups, with 55 patients in each group. Patients in the control group were treated with drugs, and those in the study group were treated with auricular acupoint bean-embedding therapy. Before and after treatment, dizziness handicap inventory (DHI) score, activities-specific balance confidence (ABC) score, and Hamilton Anxiety Scale (HAMA) score were compared between the two groups. The latency and amplitude of sympathetic skin response (SSR) were measured.Results:After 15 days of treatment, DHI scores of all dimensions in the study group were significantly lower than those in the control group ( t = 16.13-20.62, all P < 0.05). ABC score in the study group was significantly higher than that in the control group [(87.90 ± 6.01) points vs. (80.55 ± 8.73) points, t = 3.10, P < 0.05). HAMA score in the study group was significantly lower than that in the control group [(7.85 ± 1.06) points vs. (13.30 ± 2.49) points, t = 8.43, P < 0.001]. The initial latency value of SSR in the study group was significantly higher than that in the control group [(1.95 ± 0.27) ms vs. (1.67 ± 0.21) ms, and the amplitude of SSR in the study group was significantly lower than that in the control group [(1.49 ± 0.15) mV vs. (1.70 ± 0.22) mV, t = 4.73, 4.04, both P < 0.001]. The incidence of adverse reactions in the study group was significantly lower than that in the control group [0.0% (0/55) vs. 10.9% (6/55), χ2 = 4.40, P < 0.05]. Conclusion:Auricular acupoint bean-embedding therapy can effectively alleviate the symptoms of dizziness after successful reduction of BPPV, improve patient's psychological status and autonomic nerve function, which is worthy of clinical promotion.

Strategies for improving the scientific research ability of trainees in standardized residency training of dermatology / 中华医学教育探索杂志

In this paper, combined with the characteristics of rotational learning of dermatologists, we have taken various measures such as improving the scientific research training system, strengthening the awareness of scientific research, optimizing the teaching mode, adding new assessment mechanism, hand-in-hand teaching and other measures to realize the close combination of clinical and scientific research, and lay a solid foundation for the cultivation of high-quality innovative talents in dermatology.

Analysis of volatile constituents and chemical relationship of cultivated and wild Angelica dahurica / 南方医科大学学报

OBJECTIVE@#To determine the volatile constituents and their contents in the roots of 5 cultivated Angelica dahurica and one wild A. dahurica and analyze the chemical relationship among the plants of A. dahurica.@*METHODS@#The essential oil was extracted from the roots of 5 cultivated plants of Angelica dahurica and one wild A. dahurica by water steam distillation. Gas chromatography-mass spectrometry (GC-MS) was used to separate and identify all the volatile oil components in the extracts, and their relative contents were calculated with area normalization method. We also conducted clustering analysis and principal component analysis of the volatile oil components.@*RESULTS@#We identified a total of 81 compounds from the roots of the 6 plants of Angelica dahurica, including 27 in Chuanbaizhi (Angelica dahurica cv. 'Hangbaizhi'), 34 in Hangbaizhi (Angelica dahurica cv. 'Hangbaizhi'), 24 in Qibaizhi (Angelica dahurica cv. 'Qibaizhi'), 32 in Yubaizhi (Angelica dahurica cv.'Qibaizhi'), 28 in Bobahizhi (Angelica dahurica cv.'Qibaizhi'), and 34 in Xinganbaizhi (Angelica dahuirca). These compounds included, in the order of their relative contents (from high to low), alkanes, olefins, esters, organic acids and alcohols. Among the common components found in the roots of all the plants of A. dahurica, nonylcyclopropane, cyclododecane and hexadecanoic acid were identified as the volatile oil components that showed the highest relative contents. Clustering analysis of the volatile oil components showed that wild Angelica dahurica (Xing'anbaizhi) and the 5 cultivated Angelica dahurica (Chuanbaizhi, Hangbaizhi, Qibaizhi, Yubaizhi, Bobaizhi) could be divided into two groups, and the cultivated Angelica dahurica could be divided into two subgroups: Chuanbaizhi, Yubaizhi and Hangbahizhi were clustered in one subgroup, and Qibaizhi and Bobaizhi in another. The results of principal component analysis was consistent with those of clustering analysis.@*CONCLUSION@#The main volatile oil components and their contents vary among the 6 plants of A. dahurica. Nonylcyclopropane, cyclododecane and hexadecanoic acid are the most abundant volatile oil components in all the plants of A. dahurica, which can be divided into two clusters.

Analysis of the associations of chemokine receptors expression on circulating Tfh2 and Th2 cells with sIgE level and disease severity in patients with AR / 中华耳鼻咽喉头颈外科杂志

Objective: To clarify the chemotactic characteristics of type 2 helper T cells (Th2 cells) and type 2 follicular helper T cells (Tfh2 cells) in peripheral blood of patients with allergic rhinitis (AR), and to explore the associations between the chemokine receptors expression and the levels of antigen-specific IgE (sIgE) and the severity of the disease. Methods: The peripheral blood mononuclear cells of 41 patients with AR (20 males and 21 females, aged 35.0 (24.5, 47.0) years) and 42 healthy controls (24 males and 18 females, aged 35.0 (24.8, 46.5) years) treated in Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from April 2017 to February 2018 were isolated. The expressions of chemokine receptor (CCR)2, CCR3, CCR4, CCR5, CCR7, CCR8, chemokine C-X3-C-motif receptor 1 (CX3CR1) and C-X-C motif receptor 4 (CXCR4) in Th2 and Tfh2 cells were explored by fluorescence activated cell sorting (FACs). The relationship between the expression of these chemokine receptors in Th2 cells and Tfh2 cells and the levels of serum sIgE and the scores of visual analogue scale (VAS) was analyzed. Graphpad prism 7.0 software was used for statistical analysis. Results: The significant differences in chemotactic characteristics between Th2 cells and Tfh2 cells in the control group were found: Th2 cells highly expressed chemokine receptors CCR2, CCR3, CCR5, CCR8 and CX3CR1, while Tfh2 cells highly expressed immune cell homing chemokine receptors CCR7 and CXCR4. AR patients, compared to the control, expressed higher levels of CCR2, CCR5 and CX3CR1 on peripheral Th2 cells(all P<0.01). At the same time, the proportion of CCR2+and CCR5+Th2 cells was positively correlated with VAS score (r value was 0.58 and 0.61, respectively, both P<0.01). In AR patients, higher expression levels of CCR7 on Tfh2 cells were detected (P<0.01), and the proportion of CCR7+Tfh2 cells was positively correlated with the level of serum sIgE (r=0.51, P<0.01). Conclusion: The percentage of CCR2+ and CCR5+ Th2 cells in peripheral of AR patients can reflect the severity of AR to some extent, while the percentage of CCR7+ Tfh2 cells is positively correlated with the level of serum sIgE.

Application effects of bundle nursing of citric acid extracorporeal anticoagulation on continuous renal replacement therapy of severe burn patients / 中华烧伤杂志

Objective: To explore the application effects of bundle nursing of citric acid extracorporeal anticoagulation on continuous renal replacement therapy (CRRT) of severe burn patients. Methods: A non-randomized controlled study was conducted. Forty-six patients who met the inclusion criteria and received regular nursing of citric acid extracorporeal anticoagulation during CRRT in the First Affiliated Hospital of Army Medical University (the Third Military Medical University) from January to December 2017 were included in regular nursing group (30 males and 16 females, aged 42.0 (38.7,47.0) years, with 201 times of CRRT performed), and 48 patients who met the inclusion criteria and received bundle nursing of citric acid extracorporeal anticoagulation during CRRT in the same hospital from January to December 2018 were included in bundle nursing group (32 males and 16 females, aged 41.0 (36.0,46.0) years, with 164 times of CRRT performed). The clinical data of all the patients in the two groups were recorded, including the length of intensive care unit (ICU) stay, total cost of treatment in ICU, cost of CRRT, unplanned ending of treatment, ending of treatment due to operation (with the rates of unplanned ending of treatment and ending of treatment due to operation calculated), times of disposable hemodialysis filter and supporting pipeline filter (hereinafter referred to as filter) with use time>24 h, times of CRRT, and lifetime of filter. For the patients in both groups who continuously received CRRT for 3 days or more from the first treatment, the prothrombin time (PT), activated partial thromboplastin time (APTT), international normalized ratio (INR), total calcium, ionic calcium (with the difference of total calcium or ionic calcium between before and after treatment calculated), creatinine, urea, β2 microglobulin, cystatin C, platelet count, mean arterial pressure, pH value, oxygenation index, bicarbonate radical, and lactic acid levels before the first treatment (hereinafter referred to as before treatment) and 3 days after the first treatment (hereinafter referred to as after 3 days of treatment). The treatment-related complications of all patients in the two groups were recorded during hospitalization. Data were statistically analyzed with independent sample t test, Mann-Whitney U test, and chi-square test. Results: Compared with those in regular nursing group, the length of ICU stay was significantly shortened (Z=-4.71, P<0.01), the total cost of treatment in ICU was significantly reduced (t=-1.39, P<0.01), the cost of CRRT had no significant change (P>0.05), the rates of unplanned ending of treatment and ending of treatment due to operation were both significantly decreased (with χ2 values of 12.20 and 17.83, respectively, P<0.01), the times of filter service time>24 h was increased significantly (Z=-5.93, P<0.01), the times of CRRT were significantly reduced (Z=-4.75, P<0.01), and the filter service life was significantly prolonged (Z=-9.24, P<0.01) among patients in bundle nursing group. Thirty-one patients in bundle nursing group and 28 patients in regular nursing group continuously received CRRT for 3 days or more from the first treatment. Before treatment, PT, APTT, and INR of patients in bundle nursing group were 24.10 (16.08, 39.20) s, 38.81 (32.32, 45.50) s, and 1.17 (1.12, 1.19), respectively, similar to 31.75 (22.99, 40.96) s, 41.82 (35.05, 48.06) s, and 1.15 (1.11, 1.19) of patients in regular nursing group (P>0.05); the levels of total calcium and ionic calcium of patients in the two groups were similar (P>0.05). After 3 days of treatment, PT, APTT, and INR of patients in bundle nursing group and regular nursing group were 29.06 (20.11, 39.46) s, 35.25 (30.06, 40.28) s, 1.13 (1.09, 1.17) and 36.51 (26.64, 42.92) s, 39.89 (34.81, 46.62) s, 1.14 (1.10, 1.18), respectively, similar to those before treatment (P>0.05); the level of ionic calcium of patients in regular nursing group was significantly higher than that before treatment (Z=-2.08, P<0.05); the levels of total calcium and ionic calcium of patients in bundle nursing group were both significantly higher than those before treatment (with Z values of -3.55 and -3.69, respectively, P<0.01); compared with those in regular nursing group, APTT of patients was significantly shorter (Z=-2.29, P<0.05), while the total calcium level of patients was significantly higher in bundle nursing group (Z=-2.26, P<0.05). The difference of total calcium between before and after treatment of patients in bundle nursing group was significantly higher than that in regular nursing group (Z=-3.15, P<0.01). The differences of ionic calcium between before and after treatment of patients in the two groups were similar (P>0.05). Before treatment, the level of β2 microglobulin of patients in bundle nursing group was significantly higher than that in regular nursing group (Z=-2.84, P<0.01), the platelet count of patients in bundle nursing group was significantly lower than that in regular nursing group (Z=-2.44, P<0.05), while the levels of creatinine, urea, cystatin C, mean arterial pressure, pH value, oxygenation index, bicarbonate radical, and lactic acid of patients in the two groups were similar (P>0.05). After 3 days of treatment, the levels of creatinine, urea, β2 microglobulin, cystatin C, pH value, bicarbonate radical, and lactic acid of patients were all significantly lower than those before treatment (with Z values of -2.10, -2.90, -3.11, -2.02, -2.34, -2.63, and -2.84, respectively, P<0.05 or P<0.01), while the levels of platelet count, oxygenation index, and mean arterial pressure of patients were all significantly higher than those before treatment in bundle nursing group (with Z values of -6.65 and -2.40, respectively, t=-9.97, P<0.05 or P<0.01); the levels of creatinine, urea, β2 microglobulin, cystatin C, platelet count, pH value, bicarbonate radical, and lactic acid of patients were all significantly lower than those before treatment (with Z values of -5.32, -2.31, -2.41, -2.21, -3.68, -2.93, -2.20, and -2.31, respectively, P<0.05 or P<0.01), while the oxygenation index and mean arterial pressure of patients were both significantly higher than those before treatment in regular nursing group (Z=-5.59, t=-7.74, P<0.01). After 3 days of treatment, compared with those in regular nursing group, the levels of creatinine, cystatin C, platelet count, oxygenation index, bicarbonate radical, and mean arterial pressure of patients were all significantly higher (with Z values of -2.93, -1.99, -6.39, -2.09, and -2.52, respectively, t=-3.28, P<0.05 or P<0.01), while the levels of urea, β2 microglobulin, pH value, and lactic acid of patients were all significantly lower (with Z values of -3.87, -2.58, -4.24, and -2.75, respectively, P<0.05 or P<0.01) in bundle nursing group. During hospitalization, there were no treatment-related bleeding events or hypernatremia related to citric acid treatment of patients in the two groups. The ratio of total calcium to ionic calcium in one patient in bundle nursing group was >2.5, but there was no manifestation of citric acid accumulation poisoning; 1 patient had hypoionic calcemia, and 1 patient had severe metabolic alkalosis. Five patients had hypoionic calcemia and 2 patients had severe metabolic alkalosis in regular nursing group. Conclusions: The implementation of bundle nursing of citric acid extracorporeal anticoagulation during CRRT for severe burn patients shortens the length of ICU stay, reduces the total cost of treatment in ICU and the occurrence of treatment-related complications, relieves the economic burden of patients, and improves the continuity and quality of treatment.

Pay attention to the diagnosis and treatment of fundus complications related to tumor immunotherapy / 中华眼底病杂志

Cancer immunotherapy refers to the therapeutic effect of controlling or eliminating tumor cells by interfering with the immune system to restore the anti-tumor immune response. Immune checkpoint inhibitor therapy that blocks programmed death -1/programmed cell death ligand-1/cytotoxic T lymphocyte-associated antigen 4 is one of the most commonly used tumor immunotherapies, with good efficacy and wide application. These drugs cause immune-related ocular complications such as uveitis, autoimmune retinopathy, and scleritis, which represent a new etiology of ocular inflammation. The ophthalmologist's grasp of the clinical characteristics of these diseases is helpful for timely diagnosis. At the same time, the ophthalmologist will work closely with the oncologist to make a comprehensive judgment based on the patient's primary tumor, survival prognosis, severity of adverse reactions related to ocular immunotherapy, and visual prognosis, and develop suitable therapeutic strategie, thereby saving the patients' vision and improving the quality of life.

Hematopoietic stem cell transplantation of non-cryopreserved sibling umbilical cord blood for major thalassaemia in children: a report of 9 cases / 中华器官移植杂志

Objective:To explore the safety and advantages of non-cryopreserved sibling umbilical cord blood hematopoietic stem cell transplantation for major thalassaemia in children.Methods:From October 2016 to June 2021, 9 patients with major beta thalassaemia received non-cryopreserved hematopoietic stem cell transplantation of sibling umbilical cord blood at Zhongshan Hospital of Xiamen University. The pretreatment scheme, the process of stem cell implantation and follow-up were analyzed and summarized.Results:Among the 9 cases, there were 5 males and 4 females with a median age of 4(2~11)years. Median level of ferritin was 2 997(1 936~5 512)μg/L. At gestational weeks 12~16, each patient's mother underwent villi testing to confirm that the donor without thalassaemia major was complete HLA-matched with the patient. All of them received an intensive conditioning regimen made up of cyclophosphamide(CTX), fludarabine and busulfan(Bu). Graft-versus-host disease(GVHD) was prevented by cyclosporine A(CSA)and mycophenolate mofetil(MMF)with or without methotrexate(MTX). Except for one failed implant, 8 cases were successfully engrafted. Median time of neutrophil implantation was 19.5(15~26)days, median time of platelet implantation 32(22~34)days and median time of erythrocyte implantation 30.5(18~37)days. Up until September 1, 2021, the median follow-up period was 27(3~59)months and the rate of successful engraftment 88.89%. There was no transplant-related mortality. Overall survival was 100% and thalassaemia-free survival 88.89%. Two patients developed grades Ⅱ skin acute GVHD(22.2%). No grade Ⅲ-Ⅳ GVHD or chronic GVHD occurred. Epstein-Barr virus infection occurred in 1 case.No infection of cytomegalovirus occurred.Conclusions:For major thalassaemia in children, stem cell transplantation of non-cryopreserved sibling cord blood is both safe and feasible with a high implantation rate and a low incidence of GVHD.

Mechanism study of Curcumin on inhibiting experimental choroidal neovascularization <i>in vitro</i> by AKT/HIF-1α/VEGF pathway / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the effect and mechanism of curcumin on inhibiting choroidal neovascularization(CNV)<i>in vitro</i>. METHODS: Human retinal pigment epithelial(ARPE-19)cells chemical hypoxia model was established by cobalt chloride(CoCl2). CCK-8 method was used to detect the effect of curcumin on the activity of ARPE-19 cells induced by CoCl2. RT-qPCR and Western blot were used to detect the expression of AKT, HIF-1α, VEGF mRNA and protein in ARPE-19 cells hypoxia model induced by CoCl2. Cell scratch test, transwell chamber migration test, transwell chamber invasion test and matrigel matrix hose lumen formation test were used to observe the effects of conditioned medium of curcumin in ARPE-19 cells on the proliferation, migration, invasion and lumen formation of human umbilical vein endothelial cells(HUVEC)in non-contact condition. RESULTS:Chemical hypoxia model of ARPE-19 cells can successfully establish by CoCl2 at 100μmol/L. CoCl2 at the final concentration of 100μmol/L can promote the expression of AKT, HIF-1α and VEGF mRNA and p-AKT, HIF-1α and VEGF protein in ARPE-19 cells. Curcumin at the final concentration of 100μmol/L can reduce the expression of AKT, HIF-1α and VEGF mRNA in ARPE-19 hypoxia model. Curcumin at the final concentration of 100μmol/L can reduce the expression of AKT, HIF -1α and VEGF proteins in ARPE-19 hypoxia model. The conditioned medium of low(6.25μmol/L), medium(25μmol/L)and high dose(100μmol/L)curcumin in ARPE-19 cells can significantly inhibit the level migration of HUVEC. The conditioned medium in high dose group can significantly inhibit the vertical migration and cell invasion of HUVEC. The conditioned medium of middle and high dose curcumin in ARPE-19 cells can inhibit the lumen formation of HUVEC. CONCLUSION:Curcumin at 100μmol/L can protect ARPE-19 cells from hypoxia induced by CoCl2. Curcumin can inhibit the formation of blood vessels at the cellular level.

Protective effect of excretory-secretory proteins from Trichinella spiralis muscle larvae against myocardial injury in septic mice / 南方医科大学学报

OBJECTIVE@#To evaluate the protective effect of excretory-secretory proteins from Trichinella spiralis muscle larvae (Ts-MES) on sepsis-induced myocardial injury in mice.@*METHODS@#Eighty male BALB/C mice were randomized equally into sham-operated group, myocardial injury group, Ts-MES treatment group and dexamethasone treatment group. In the latter 3 groups, sepsis-induced myocardial injury models were established by cecal ligation and perforation; the sham operation was performed by exposure of the cecum without ligation or perforation. Forty minutes after the operation, the mice were given intraperitoneal injections 150 μL PBS, 20 μg TS-MES or 0.3 mg/kg dexamethasone as indicated. At 12 h after the operation, 6 mice were randomly selected from each group for echocardiography, and 8 mice were used for observing the survival rate within 72 h. The remaining 6 mice were examined for myocardial pathologies with HE staining and serum levels of NTPro-BNP and cTnI with ELISA; the expressions of TNF-α, IL-6, IL-10 and TGF-β in the serum and myocardial tissue were detected using ELISA and qRT-PCR.@*RESULTS@#Compared with the sham-operated mice, the septic mice showed significantly decreased cardiac function indexes (LVEF, LVFS, and E/A) with lowered survival rate within 72 h (P < 0.001) and significantly higher myocardial injury scores and serum levels of NTPro-BNP and cTnI (P < 0.01). Treatment with TS-MES significantly improved the cardiac function and 72-h survival rate (P < 0.05) and lowered the myocardial injury scores and serum levels of NTPro-BNP and cTnI (P < 0.05) in the septic mice. Compared with the sham-operated mice, the septic mice had obviously increased TNF-α and IL-6 levels in the serum and myocardial tissue (P < 0.001), which were significantly lowered by treatment with TS-MES (P < 0.05). TS-MES and dexamethasone both increased the levels of IL-10 and TGF-β in the septic mice, but the changes were significant only in TS-MES-treated mice (P < 0.05).@*CONCLUSION@#Ts-MES are capable of protecting against myocardial injury in septic mice by reducing the production of pro-inflammatory cytokines and enhancing the levels of regulatory cytokines.

Mechanism of Arnebia euchroma Against Melanoma： An Exploration Based on Network Pharmacology and Experimental Verification / 中国实验方剂学杂志

ObjectiveTo preliminarily predict the active components， action targets， and signaling pathways of Arnebia euchroma in the treatment of melanoma based on network pharmacology and molecular docking， and to verify its possible mechanism of action in in vitro experiments. MethodThe active components and related targets of A. euchroma were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）SwissTargetPrediction and literature， and the targets related to melanoma from the GeneCards， Online Mendelian Inheritance in Man （OMIM）， and Comparative Toxicogenomics Database （CTD）. Following the construction of the protein-protein interaction （PPI） network of active components and related targets of A. euchroma and melanoma-related targets using STRING， Cytoscape 3.8.2 was used for screening and analyzing the nodes in the network of A. euchroma against melanoma. The intersections were subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analysis using DAVID 6.8. Acetyl alkannin， the active component in A. euchroma， was docked to the target by AutoDock Vina 1.1.2. The in vitro experiments were then carried out to verify the anti-melanoma effect of A. euchroma. ResultA total of 271 common targets of A. euchroma and melanoma were harvested， among which 23 were key targets， including matrix metalloproteinase-9 （MMP-9） and Janus kinase 2 （JAK2）. As revealed by KEGG enrichment analysis， A. euchroma mainly acted on Janus kinase/signal transduction and activator of transcription （JAK/STAT）， tyrosine kinase receptor （ErbB）， and vascular endothelial growth factor （VEGF） signaling pathways to resist melanoma. According to molecular docking， acetyl alkannin exhibited a good docking activity with JAK2， STAT3， VEGF， MMP-9， and E-cadherin receptors. The results of Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） showed that acetyl alkannin at different doses inhibited the protein and gene expression of JAK2， STAT3， VEGF， MMP-9， and E-cadherin in A375 cells （P<0.05）. ConclusionA. euchroma alleviates melanoma via multiple targets and multiple pathways， and it may exert the therapeutic effects by affecting the expression of such key target proteins as JAK2， STAT3， VEGF， MMP-9， and E-cadherin and inhibiting the invasion and metastasis of melanoma cells. This study has provided an experimental basis for the treatment of tumor with A. euchroma.

Analysis of muscle activation in children and adolescents with severe cerebral palsy / Análise da ativação muscular em crianças e adolescentes com paralisia cerebral grave

Abstract Introduction: Children and adolescents with spastic quadriparesis have a worse selective motor control, and studies with this population are still very scarce. The same is true with scientific evidence of one of the methods most used as a physiotherapeutic treatment in this population, the Bobath Concept. Objective: To evaluate spine erector muscles activation, gluteus medius and gluteus maximus, through the handling of the Bobath Concept and the sustained kneeling posture in subjects with a diagnosis of severe cerebral palsy; and to compare muscle activation with a reference group, in order to increase the reliability of this study. Methods: A cross-sectional study was carried out with 38 children and adolescents with cerebral palsy, classified by GMFCS at levels IV and V, and 20 healthy participants, aged between 3 and 18 years. They were submitted to the handling of the Bobath Concept and to the sustained kneeling posture, with muscle activation obtained by electromyography. Results: We observed significant muscle activation during handling in side-sitting, with weight transfer and without the help of another therapist, and in the sustained kneeling posture, for the erector of the spine and gluteus medius. Conclusion: The evidence from this study suggests that both the handling in side-sitting and the sustained kneeling posture cause significant muscle activation in the erector of the spine and gluteus medius for severe quadriparesis subjects, GMFCS IV and V, which can contribute to the improvement of postural control and decision-making in physical therapy practice.

Resumo Introdução: Crianças e adolescentes com quadriparesia espástica apresentam pior controle motor seletivo e estudos com essa população ainda são muito escassos. O mesmo ocorre com as evidências científicas de um dos métodos mais utilizados como tratamento fisioterapêutico nessa população, o Conceito Bobath. Objetivo: Avaliar a ativação dos músculos eretores da coluna, glúteo médio e glúteo máximo por meio do manuseio do Conceito Bobath e da postura ajoelhada sustentada, em indivíduos com diagnóstico de paralisia cerebral grave; e comparar a ativação muscular com um grupo de referência, a fim de aumentar a confiabilidade deste estudo. Métodos: Realizou-se um estudo transversal com 38 crianças e adolescentes com paralisia cerebral, classificados pelo GMFCS nos níveis IV e V, e 20 participantes saudáveis com idade entre 3 e 18 anos. Eles foram submetidos ao manuseio do Conceito Bobath e à postura ajoelhada sustentada, com ativação muscular obtida por eletromiografia. Resultados: Observou-se ativação muscular significativa durante o manuseio na posição sentada de lado, com transferência de peso e sem auxílio de outro terapeuta, e na postura ajoelhada sustentada para o eretor da coluna e glúteo médio. Conclusão: As evidências deste estudo sugerem que tanto o manuseio na posição sentada de lado quanto a sustentação da postura ajoelhada causam significativa ativação muscular no eretor da coluna e glúteo médio para indivíduos com quadriparesia grave, GMFCS IV e V, o que pode contribuir para a melhora do controle postural e tomada de decisão na prática fisioterapêutica.

Incidence of extrauterine growth retardation and its risk factors in very preterm infants during hospitalization: a multicenter prospective study / 中国当代儿科杂志

OBJECTIVES@#To investigate the incidence of extrauterine growth retardation (EUGR) and its risk factors in very preterm infants (VPIs) during hospitalization in China.@*METHODS@#A prospective multicenter study was performed on the medical data of 2 514 VPIs who were hospitalized in the department of neonatology in 28 hospitals from 7 areas of China between September 2019 and December 2020. According to the presence or absence of EUGR based on the evaluation of body weight at the corrected gestational age of 36 weeks or at discharge, the VPIs were classified to two groups: EUGR group (n=1 189) and non-EUGR (n=1 325). The clinical features were compared between the two groups, and the incidence of EUGR and risk factors for EUGR were examined.@*RESULTS@#The incidence of EUGR was 47.30% (1 189/2 514) evaluated by weight. The multivariate logistic regression analysis showed that higher weight growth velocity after regaining birth weight and higher cumulative calorie intake during the first week of hospitalization were protective factors against EUGR (P<0.05), while small-for-gestational-age birth, prolonged time to the initiation of total enteral feeding, prolonged cumulative fasting time, lower breast milk intake before starting human milk fortifiers, prolonged time to the initiation of full fortified feeding, and moderate-to-severe bronchopulmonary dysplasia were risk factors for EUGR (P<0.05).@*CONCLUSIONS@#It is crucial to reduce the incidence of EUGR by achieving total enteral feeding as early as possible, strengthening breastfeeding, increasing calorie intake in the first week after birth, improving the velocity of weight gain, and preventing moderate-severe bronchopulmonary dysplasia in VPIs.

Clinical treatment outcomes and their changes in extremely preterm twins: a multicenter retrospective study in Guangdong Province, China / 中国当代儿科杂志

OBJECTIVES@#To investigate the clinical treatment outcomes and the changes of the outcomes over time in extremely preterm twins in Guangdong Province, China.@*METHODS@#A retrospective analysis was performed for 269 pairs of extremely preterm twins with a gestational age of <28 weeks who were admitted to the department of neonatology in 26 grade A tertiary hospitals in Guangdong Province from January 2008 to December 2017. According to the admission time, they were divided into two groups: 2008-2012 and 2013-2017. Besides, each pair of twins was divided into the heavier infant and the lighter infant subgroups according to birth weight. The perinatal data of mothers and hospitalization data of neonates were collected. The survival rate of twins and the incidence rate of complications were compared between the 2008-2012 and 2013-2017 groups.@*RESULTS@#Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of severe asphyxia and smaller head circumference at birth (P<0.05). The mortality rates of both of the twins, the heavier infant of the twins, and the lighter infant of the twins were lower in the 2013-2017 group compared with the 2008-2012 group (P<0.05). Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of pulmonary hemorrhage, patent ductus arteriosus (PDA), periventricular-intraventricular hemorrhage (P-IVH), and neonatal respiratory distress syndrome (NRDS) and a higher incidence rate of bronchopulmonary dysplasia (P<0.05).@*CONCLUSIONS@#There is a significant increase in the survival rate over time in extremely preterm twins with a gestational age of <28 weeks in the 26 grade A tertiary hospitals in Guangdong Province. The incidences of severe asphyxia, pulmonary hemorrhage, PDA, P-IVH, and NRDS decrease in both the heavier and lighter infants of the twins, but the incidence of bronchopulmonary dysplasia increases. With the improvement of diagnosis and treatment, the multidisciplinary collaboration between different fields of fetal medicine including prenatal diagnosis, obstetrics, and neonatology is needed in the future to jointly develop management strategies for twin pregnancy.

Icariin improves renal interstitial fibrosis in a rat model of chronic renal failure by regulating mitochondrial dynamics / 中国中药杂志

This study aims to explore the effect of icariin(ICA) on mitochondrial dynamics in a rat model of chronic renal failure(CRF) and to investigate the molecular mechanism of ICA against renal interstitial fibrosis(RIF). CRF was induced in male Sprague-Dawley(SD) rats with 5/6(ablation and infarction, A/I) surgery(right kidney ablation and 2/3 infarction of the left kidney). Four weeks after surgery, the model rats were randomized into the following groups: 5/6(A/I) group, 5/6(A/I)+low-dose ICA group, and 5/6(A/I)+high-dose ICA group. Another 12 rats that received sham operation were randomly classified into 2 groups: sham group and sham+ICAH group. Eight weeks after treatment, the expression of collagen-Ⅰ(Col-Ⅰ), collagen-Ⅲ(Col-Ⅲ), mitochondrial dynamics-related proteins(p-Drp1 S616, p-Drp1 S637, Mfn1, Mfn2), and mitochondrial function-related proteins(TFAM, ATP6) in the remnant kidney tissues was detected by Western blot. The expression of α-smooth muscle actin(α-SMA) was examined by immunohistochemical(IHC) staining. The NRK-52 E cells, a rat proximal renal tubular epithelial cell line, were cultured in vitro and treated with ICA of different concentration. Cell viability was detected by CCK-8 assay. In NRK-52 E cells stimulated with 20 ng·mL~(-1) TGF-β1 for 24 h, the effect of ICA on fibronectin(Fn), connective tissue growth factor(CTGF), p-Drp1 S616, p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 was detected by Western blot, and the ATP content and the mitochondrial morphology were determined. The 20 ng·mL~(-1) TGF-β1-stimulated NRK-52 E cells were treated with or without 5 μmol·L~(-1) ICA+10 μmol·L~(-1) mitochondrial fusion promoter M1(MFP-M1) for 24 h and the expression of fibrosis markers Fn and CTGF was detected by Western blot. Western blot result showed that the levels of Col-Ⅰ, Col-Ⅲ, and p-Drp1 S616 were increased and the levels of p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 were decreased in 5/6(A/I) group compared with those in the sham group. The levels of Col-Ⅰ, Col-Ⅲ, and p-Drp1 S616 were significantly lower and the levels of p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6 were significantly higher in ICA groups than that in 5/6(A/I) group. IHC staining demonstrated that for the expression of α-SMA in the renal interstitium was higher in the 5/6(A/I) group than in the sham group and that the expression in the ICA groups was significantly lower than that in the 5/6(A/I) group. Furthermore, the improvement in the fibrosis, mitochondrial dynamics, and mitochondrial function were particularly prominent in rats receiving the high dose of ICA. The in vitro experiment revealed that ICA dose-dependently inhibited the increase of Fn, CTGF, and p-Drp1 S616, increased p-Drp1 S637, Mfn1, Mfn2, TFAM, and ATP6, elevated ATP content, and improved mitochondrial morphology of NRK-52 E cells stimulated by TGF-β1. ICA combined with MFP-M1 further down-regulated the expression of Fn and CTGF in NRK-52 E cells stimulated by TGF-β1 compared with ICA alone. In conclusion, ICA attenuated RIF of CRF by improving mitochondrial dynamics.

Identification of chemical constituents of Xiaochengqi Decoction by UPLC-Q-TOF/Fast DDA combined with UNIFI software / 中国中药杂志

Based on the combination of ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF) and Waters UNIFI software, the chemical constituents of the classic prescription Xiaochengqi Decoction were qualitatively analyzed and identified. The UPLC conditions are as follows: Acquity HSS T3 reverse phase column(2.1 mm ×100 mm, 1.8 μm), column temperature of 30 ℃, mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B), and flow rate of 0.3 mL·min~(-1). High-resolution MS data of Xiaochengqi Decoction were collected in ESI~(+/-) modes by Fast DDA. The structures of the chemical constituents were tentatively characterized or identified by UNIFI software according to the retention time of reference standards and characteristic fragment ions in MS profile, and literature data. A total of 233 components in Xiaochengqi Decoction were identified, with 93 from wine-processed Rhei Radix et Rhizoma, 104 from bran-processed Aurantii Fructus Immaturus, and 36 from ginger-processed Magnoliae Officinalis Cortex. These 233 components included anthraquinones, flavonoids, lignans, alkaloids, coumarins, and phenylethanoid glycosides. The result provided experimental evidence for the further study on establishment of quality standard and product development of the formula.

Risk factors of bronchopulmonary dysplasia in very preterm infants: a national multicenter study / 中国小儿急救医学

Objective:To analyze the risk factors of bronchopulmonary dysplasia(BPD)in very preterm infants(VPI), and to provide scientific basis for the prevention and treatment of BPD in VPI.Methods:A prospective multicenter study was designed to collect the clinical data of VPI in department of neonatology of 28 hospitals in 7 regions from September 2019 to December 2020.According to the continuous oxygen dependence at 28 days after birth, VPI were divided into non BPD group and BPD group, and the risk factors of BPD in VPI were analyzed.Results:A total of 2 514 cases of VPI including 1 364 cases without BPD and 1 150 cases with BPD were enrolled.The incidence of BPD was 45.7%.The smaller the gestational age and weight, the higher the incidence of BPD( P<0.001). Compared with non BPD group, the average birth age, weight and cesarean section rate in BPD group were lower, and the incidence of male infants, small for gestational age and 5-minute apgar score≤7 were higher( P<0.01). In BPD group, the incidences of neonatal respiratory distress syndrome(NRDS), hemodynamically significant patent ductus arteriosus, retinopathy of prematurity, feeding intolerance, extrauterine growth restriction, grade Ⅲ~Ⅳ intracranial hemorrhage, anemia, early-onset and late-onset sepsis, nosocomial infection, parenteral nutrition-associated cholestasis were higher( P<0.05), the use of pulmonary surfactant(PS), postnatal hormone exposure, anemia and blood transfusion were also higher, and the time of invasive and non-invasive mechanical ventilation, oxygen use and total hospital stay were longer( P<0.001). The time of starting enteral nutrition, cumulative fasting days, days of reaching total enteral nutrition, days of continuous parenteral nutrition, days of reaching 110 kcal/(kg·d) total calorie, days of reaching 110 kcal/(kg·d) oral calorie were longer and the breastfeeding rate was lower in BPD group than those in non BPD group( P<0.001). The cumulative doses of amino acid and fat emulsion during the first week of hospitalization were higher in BPD group( P<0.001). Multivariate Logistic regression analysis showed that NRDS, invasive mechanical ventilation, age of reaching total enteral nutrition, anemia and blood transfusion were the independent risk factors for BPD in VPI, and older gestational age was the protective factor for BPD. Conclusion:Strengthening perinatal management, avoiding premature delivery and severe NRDS, shortening the time of invasive mechanical ventilation, paying attention to enteral nutrition management, reaching whole intestinal feeding as soon as possible, and strictly mastering the indications of blood transfusion are very important to reduce the incidence of BPD in VPI.