ABSTRACT
Retrospective analysis was performed on 1 child with silent inactivation (SI) of asparaginase (ASNas) who was diagnosed with acute lymphoblastic leukemia (ALL) and treated in the First Affiliated Hospital, Sun Yat-Sen University in October 2019.The patient was a 9 years and 3 months old boy who was diagnosed as ALL accompanied with late bone marrow relapse.After pegylated Escherichia coli-Asparaginase (PEG-ASNase) was given, he did not have the expected treatment-related adverse reactions, including hyperammonemia, hypofibrinogenemia, and the low activation of antithrombin Ⅲ (ATⅢ). The plasma asparagine (ASN) concentration failed to meet the depletion criteria and the ASNase activity was 64.5 U/L.Therefore, the SI of ASNase was confirmed.Erwinase was used to replace PEG-ASNase, the lowest level of ATⅢ was 33%, and the lowest level of fibrinogen was 1.20 g/L.Hyperammonemia and decreased ASN were also observed, and the ASNase activity was 1 813.0 U/L.All the above suggested that when, SI occurred, the replacement by Erwinase was effective.The ASNase activity should be monitored in ALL patients who were treated with ASNase.Monitoring the treatment-related adverse reactions such as hyperammonia and coagulation disorders closely has important implications to the SI of ASNase when the detection of ASNase activity was unavailable.
ABSTRACT
Symptomatology and medical history taking are the focuses of diagnostics study. With the improvement of teaching and learning, students can basically master the standard process of clinical inquiry of chief complaints to family history efficiently. However, due to the lack of systematic learning of clinical diseases, it's difficult for most students to form a targeted, logical and speculative inquiry thinking pattern, so they are unlikely to write a medical history of present illness that reflects their thinking on disease inclusion and exclusion. In this study, we come up with the characteristics and defects of symptomatology teaching at present, and put forward a new idea of symptomatology teaching by introducing clinical manifestations of diseases and examples of inquiry.
ABSTRACT
Oral squamous cell carcinoma (OSCC) has a high incidence of metastasis. Tumour immunotherapy targeting PD-L1 or PD-1 has been revolutionary; however, only a few patients with OSCC respond to this treatment. Therefore, it is essential to gain insights into the molecular mechanisms underlying the growth and metastasis of OSCC. In this study, we analysed the expression levels of protein kinase D3 (PKD3) and PD-L1 and their correlation with the expression of mesenchymal and epithelial markers. We found that the expression of PKD3 and PD-L1 in OSCC cells and tissues was significantly increased, which correlated positively with that of mesenchymal markers but negatively with that of epithelial markers. Silencing PKD3 significantly inhibited the growth, metastasis and invasion of OSCC cells, while its overexpression promoted these processes. Our further analyses revealed that there was positive feedback regulation between PKD3 and PD-L1, which could drive EMT of OSCC cells via the ERK/STAT1/3 pathway, thereby promoting tumour growth and metastasis. Furthermore, silencing PKD3 significantly inhibited the expression of PD-L1, and lymph node metastasis of OSCC was investigated with a mouse footpad xenograft model. Thus, our findings provide a theoretical basis for targeting PKD3 as an alternative method to block EMT for regulating PD-L1 expression and inhibiting OSCC growth and metastasis.
Subject(s)
Animals , Humans , Mice , B7-H1 Antigen/metabolism , Carcinoma, Squamous Cell , Cell Line, Tumor , Feedback , Head and Neck Neoplasms , Mouth Neoplasms , Protein Kinase C , STAT1 Transcription Factor , Squamous Cell Carcinoma of Head and NeckABSTRACT
[Objective]To explore the effect and the possible mechanism of the proteasome inhibitor MG132 on acute T lympho?blastic leukemia cells.[Methods]The influence of different concentrations of MG132 in the viability and proliferation of CCRF-CEM was measured by MTS. Apoptosis rates of CCRF-CEM treated by MG132 were determined by flow cytometry. After being exposed to MG132,the protein levels of FOXO3a in cytoplasm and nucleus were analyzed by Western blotting. qRT-PCR was applied to detect the mRNA of FOXO3a and Puma in cells treated by MG132. Then CCRF-CEM was stably transfected with antisense FOXO3a using Lentivirus infection. We further investigated the effects of MG132 in FOXO3a-shRNA cells and elucidated the mechanisms of FOXO3a and Puma.[Results]MG132 inhibits the proliferation of CCRF-CEM,but has no cytotoxicity in peripheral blood mononu?clear cells(PBMC). Cellular apoptosis was induced in cells treated with MG132. At mRNA level,MG132 had no influence on FOXO3a,but increased the expression of Puma. However,MG132 promoted the expression of both FOXO3a and Puma at protein level. Interestingly,the expression of FOXO3a increased very little in cytoplasm. In FOXO3a-shRNA cells the expression of FOXO3a and Puma decreased at protein level. FOXO3a's knockdown attenuated the proliferation inhibition mediated by MG132.[Conclusion]MG132 inhibits the proliferation and promotes to apoptosis of CCRF-CEM. One of the mechanism is that MG132 inhib? its the degradation of FOXO3a,and then activates FOXO3a/Puma pathway.
ABSTRACT
Objective To explore the effect of Flavokawain B on the proliferation and apoptosis of acute T lymphoblastic leukemia(T -ALL)cells and its preliminary mechanism.Methods After the T -ALL cell lines CEM-C7(sensitive to glucocorticoids)and MOLT -4(resistant to glucocorticoids)cells were treated with different concentrations of Flavokawain B,the influence of Flavokawain B on the growth rate and doubling time of CEM-C7 and MOLT -4 cells was observed by 3 -(4,5 -dimethylthiazol -2 -yl)-5 -(3 -carboxymethoxyphenyl)-2 -(4 -sulfophenyl)-2H -tetrazolium(MTS)assay,and apoptosis was analyzed by using flow cytometry.Furthermore,Wes-tern blot assay was used to detect the expressions of Bim,Bcl -2 and cleaved Caspase -9.At last,the expressions of Bim and Bcl -2 in clinical T -ALL patient samples were also detected by using Western blot assay.Results MTS as-say showed that Flavokawain B significantly inhibited the cellular proliferation of T -ALL cell lines in a dose and time dependent manner(P <0.01 ).Flow cytometry findings revealed that Flavokawain B significantly induced the apoptosis of T -ALL cells in a dose -dependent manner(P <0.001 ).Western blot results indicated that Flavokawain B in-creased the expression of Bim and cleaved Caspase -9,and decreased the expression of Bcl -2 in T -ALL cell lines, which increased Bim and decreased Bcl -2 in clinical T -ALL patients samples,both in a dose -dependent manner. Conclusions Flavokawain B can inhibit the proliferation and induce the apoptosis of T -ALL cells by up -regulating the expression of Bim and down -regulating the expression of Bcl -2 and activating Caspase -9,whether resistant to glu-cocorticoids or not.
ABSTRACT
Background Retinal microglia (RMG) plays an important role in the pathogenesis of retinal degenerative diseases,while chemokine CX3CL1 participates in the regulation of steady-state of microglia.It has been determined that bone marrow-derived mesenchymal stem cells (BMSCs) have a remarkable role to modulate the immune response and protect the central nervous system through the release of soluble factors in a paracrine fashion and further affect the functional behavior of cells.However,whether BMSCs are able to interact with RMG and activate related signaling pathway for the maintaining of homeostasis in the retina is still unclear.Objective The aim of this study was to investigate the interaction between BMSCs and lipopolysaccharide (LPS)-activated RMG in vitro,and dissect the effects of CX3CL1/CX3CR1 signaling pathway on the biological behavior of BMSCs and RMG.Methods RMG was isolated from SD rats,cultured with mixed culture of retinal glial cells and purified by shaking.The cells were identified by detecting the expression of CD111b,Iba1 and glutamamine synthetase (GS) with indirect immunofluorescence assay.LPS (1 mg/ml,2 μl) was added in the medium for 24 hours to stimulate RMG,and then the cells were divided into LPS control group,BMSCs group (cocultured with BMSCs for 24 hours) and CB-BMSCs group (cocultured with CX3CL1-blocking-BMSCs for 24 hours).The cells without LPS stimulation served as the blank control group.The functions of RMG,including the release content of tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β),the proliferation,phagocytosis,and migration of RMG were examined.Results RMG was successfully isolated and harvested from SD rats by using mixed culture of retinal glial cells and purified by shaking.CD11b and Iba1 showed the positive expression with the green fluorescence in the cells and GS was absent.The contents of TNF-αt in the cell supernatant were (2.55 ±0.97) ng/ml,(24.91 ±3.07) ng/ml,(20.38 ±2.97) ng/ml and (24.90 ± 1.88) ng/ml in the blank control group,LPS control group,BMSCs group and CB-BMSCs group,respectively,showing a significant difference among the groups (F=119.90,P＜0.05).The contents of IL-1 β in the cell supernatant were (1.12±0.36) ng/ml,(10.40±2.76) ng/ml,(7.00± 1.75) ng/ml and (9.55 ± 1.11) ng/ml in the blank control group,LPS control group,BMSCs group and CB-BMSCs group,respectively,showing a significant difference among the groups(F =34.96,P＜0.05).The secretory volume of TNF-α and IL-1 β were evidently lower in the BMSCs group than those in the LPS control group (both at P＜0.05),and no significant differences were found in the secretory volume of TNF-α and IL-1β between CB-BMSCs group and LPS control group (both at P＞0.05).The proliferative rate of RMG was lower in the BMSCs group than that in the LPS control group (P＜0.05),while there was no statistical difference between BMSCs group and CB-BMSCs group (P＞0.05).The mean fluorescence intensity (MFI) and the number of migrated RMG were considerably different among the four groups (F=70.55,15.49,both at P＜0.05),and those in the BMSCs group were significantly increased in comparison with the LPS control group (both at P＜0.05),while there was no significant difference between CB-BMSCs group and LPS control group (both at P＞0.05).Conclusions BMSCs could suppress the proliferation of LPS-activated RMG.Moreover,BMSCs might inhibit proinflammatory cytokines releasing,enhance phagocytosis and migration capabilities of RMG via CX3CL1/CX3CR1 signaling pathway.
ABSTRACT
Objective To explore the prognostic significance in monitoring minimal residual disease (MRD) in childhood acute lymphoblastic leukemia (ALL) by a simple method,and to detect cloned immunoglobulin H (IgH) and T cell receptor γ (TCRγ) gene rearrangements by using multiplex polymerase chain reaction (PCR) and automated fragment analysis.Methods Bone marrow samples were collected from 86 newly diagnosed cases of childhood ALL at the Department of Pediatrics,the First Affiliated Hospital of Sun Yat-Sen University,from May of 2009 to August of 2013.IgH and TCRγ gene rearrangements were amplified by qualitative multiplex PCR.The clonality of PCR production was analyzed by GENEMAPPERID software.Only those carried monoclonal IgH/TCRγ on diagnosis were arranged to monitor MRD.Detectable monoclonal IgH/TCRγby the end of induction was defined as MRD positive.All patients were treated with GD2008 ALL protocol.Clinical data of all newly-diagnosed ALL patients in the corresponding period were reviewed.The final follow-up on May 31,2014.Survival rates and event free survival (EFS) curves were estimated by the Kaplan-Meier,and compared by using the log-rank test.Results The percent age of 94.2 (81/86 cases) patients was at least 1 marker positive.Subsequent MRD was monitored in 79 cases.The median follow-up time was 20 months (9-61 months).By the end of induction,20 cases were MRD positive and 59 cases were M RD negative,and the 3-year EFS were 56.4％ ± 14.7％ and 94.0％ ± 3.4％ (x2 =8.563,P =0.003),respectively.According to the traditional prognostic stratification criteria,MRD was detected 65 cases in the non-high risk group:23 cases in standard risk group and 42 cases in intermediate risk group,and the difference of 3-year EFS had no statistical significance (95.3％ ±4.7％ vs 76.6％ ±9.0％,x2 =0.934,P =0.334).While using MRD by the end of induction as a risk stratification criterion,there was a statistical significant difference compared with the 3-year EFS for MRD-negative (n =52) group and MRD-positive (n =13) group (93.1％ ± 3.8％ and 59.5％ ± 16.2％,x2 =7.128,P =0.008).Conclusions It is a simple but feasible method to monitor MRD in childhood ALL by using this qualitative multiplex PCR with automated fragment analysis for monoclonal IgH/TCRγ gene rearrangements.MRD by the end of induction can be used as a more accurate risk stratification criterion than the traditional one.It is worth of further research.
ABSTRACT
Objective For further improving the prognosis of childhood acute promyelocytic leukemia (APL) in China,the treatment efficacies, outcomes and costs of protocols for childhood APL between in developed countries and in our hospital were compared. Methods 30 cases aged ＜15 years were diagnosed according to the FAB classification and detection of PML-RARα rearrangement. From December 1999 to September 2004,sixteen eligible children were treated with an intensive in-house protocol including high-dose Ara-C and anthracycline for post remission treatment. From September 2004 to January 2008,14 cases enrolled were treated with a less intensive protocol modified from the PETHEMA LPA99. Results The 3.5 years EFS was 37.5 % (s-x=0.121) for total 16 patients on in-house protocol. Six patients (37.5 %) abandoned treatment,2 died of intracranial hemorrhage at diagnosis (6.3 %) and sepsis in remission (6.3 %),respectively,and 2 relapsed (12.5 %). The 14 cases treated with modified PETHEMA had a 3.5 years EFS of 79.6 % (s-x=0.136). One died of intracranial hemorrhage at diagnosis (7.1 %) and 1 relapsed (7.1 %). Patients on modified PETHEMA had a significantly higher EFS (P=0.012),lower frequency of sepsis during treatment (7.7 % vs 77.8 %; P=0.0015) and lower hospitalization cost (median,RMB 35 200 vs 150 000; P ＜0.0001) than those on in-house protocol. Conclusion Treatment with the less intensive protocol based on the PETHEMA LPA99 study for childhood APL successively reduced complication of chemotherapy and reduced hospitalization cost without increasing relapses, which led to decreases in treatment-related toxicity and treatment abandonment rate,thus improving overall outcome.
ABSTRACT
Objective To review the diagnosis of idiopathic pulmonary hemosiderosis ( IPH),and to evaluate the efficacy of maintenance therapy with dose-adjusted 6-mercaptopurine (6MP) in IPH children. Methods The diagnosis of IPH was confirmed by in-patient examination and at least 1 year follow-up to exclude secondary causes of pulmonary hemorrhage. Fifteen children met the criteria of IPH and were enrolled. The age at diagnosis was 2-13 years ( median 7 years). Prednisone was administered at 2 mg/( kg·d) for 4 weeks in acute phase of the disease followed by taper. 6MP was also started at 60 mg/( m2·d) simultaneously and continued for 3 years. Results The diagnosis was delayed in most children, which was due to the lack of initial classical manifestation of the disease. The time between the onset of symptoms and diagnosis ranged from 2 weeks to 108 months ( median 8 months) . All the patients exhibited response to the initial treatment and prednisone was successfully tapered off. Only 1 of 8 patients with relative leucopenia (3 × 109/L -6 × 109/L) on 6MP maintenance recurred while 5 of 7 others recurred (P < 0. 05) during median 6-year (range 2. 5 - 9. 5 years) follow-up. Of the latter 5 patients who recurred,4 remained recurrence-free after adjusting the dose of 6MP upwards to keep relative leucopenia. Conclusions Diagnostic delayed is still a main problem in pediatric IPH. Most IPH children in our group tolerated maintenance treatment with 6MP and achieved long-term remission, and these suggested growth retardation on long-term steroids therapy could be avoided. Because of interindividual difference in 6MP metabolism, adjusting the dose of 6MP may be necessary for treatment of IPH children and avoid under-treatment or overtreatment in some children,and thus improve the prognosis. White blood count could be a simple and useful indicator to predict clinical response in most IPH children on 6MP.
ABSTRACT
Objective To study the clinic features and the mechanism of skin damage induced by high-dose methotrexate(HD-MTX).Method Children treated with HD-MTX were enrolled in a retrospective study in which the incidence and presentations of skin damage were concluded.Mechanism of the damage was studied base on skin pathology of an animal model.Result Skin damage presented in 3～9(median 5) days after HD-MTX and usually followed by bullation and exfoliation.Twelve cases,more severe than the others,accompanied with fever and symptoms of other organs especially the respiratory tract and the intestine.Skin damage was reduced when large dose rehydration used before,during and after HD-MTX.Conclusion HD-MTX may cause scald-like skin damage directly by its prolonged toxic effect on epidermal cells.Besides extended tetrahydrofolic acid rescue and supportive treatments,anti infection is the most important in dealing this condition.
ABSTRACT
After the reform from "4+1" to "3+2",an undergraduate teaching model of clinical medicine,has been implemented in our university,higher demands are set on the training of students' abilities and creativity.This paper explored the basic medicine teaching from the perspectives of creative concepts,teaching staff,curriculum design and practice.