Objective:To investigate the risk factors and interventions for surgical failure of spinal tuberculosis (STB).Methods:A total of 317 STB patients aged from 11 to 86 years with an average age of 53.5±16.7 years, who received debridement and fusion with bone grafting from January 2013 to December 2019, were retrospectively analyzed, including 206 males and 111 females. The follow-up duration was at least 1 year. During the follow-up, any one of the following 1)-3) was defined as surgical failure, namely 1) the same tubercular lesion treated by surgery more than 2 times, 2) the number of unplanned readmissions related to tubercular lesion≥1, 3) drug-resistant STB or delayed healing, recurrent lesion with cold abscess/sinus tract, combined with other bacterial infection, or loosening of internal fixation. The other cases were regarded as "curative" cases. Patients' symptoms, medication history, auxiliary examination and surgical plan were collected for univariate analysis. Further, the potential risk factors for surgical failure were analyzed by binary Logistic regression. Failed cases were treated with etiological intervention, such as puncture pumping pus or debridement or revision. The necrosis or granulation tissue was collected and further detected by tuberculosis culture, metagenomic next-generation sequencing (mNGS) and real-time fluorescent quantitative PCR (RT-qPCR).Results:There were 27 cases with surgical failure. Abscess or sinus tract formation was developed in 17 cases, which accounted for 63% (17/27). Among these patients, there were 3 cases of resistance to isoniazid or rifampicin and 2 cases of resistance to isoniazid and rifampicin (multidrug resistance, MDR). Seventeen cases were treated by anti-tuberculosis treatment, while 14 cases by puncture drainage (or puncture catheter irrigation) and 3 cases by debridement and suturing. Seven cases with wound infection or poor healing accounted for 26% (7/27). Among them, 5 kinds of pathogens were detected, none of which showed tuberculosis drug resistance. All of them were treated by anti-infection and debridement suturing, while 2 of them were treated with internal fixation removal. Three cases (11%, 3/27) with internal fixation loosening were treated by revision surgery. There was statistically significant difference between the failed group and the cured group in involved multi-/jumping segment, history of type 2 diabetes, a history of more than three basic diseases, CRP at one week after surgery, WBC at one week after surgery, time of first dose, operation duration and intraoperative blood loss ( P<0.10). Binary Logistic regression analysis showed that multi-/jumping segment ( OR= 3.513, P=0.047), CRP at one week after surgery ( OR=1.021, P=0.005), first dose time ≥20 weeks ( OR=2.895, P=0.039), blood loss ≥800 ml ( OR=5.950, P=0.001) and more than three basic diseases involved ( OR=3.671, P=0.027) were independent risk factors for surgical failure. Conclusion:Early diagnosis, especially the diagnosis of drug-resistant STB and standardized anti-tubercular treatment, should be carried out effectively. Puncture and drainage of abscess is an effective therapy to treat the cases with abscess/sinus tract formation. Some cases involved multi-/jumping segments could be with higher risk of failure after internal fixation. Thus, they should be treated individually with emphasis on the segmental stability reconstruction.

Objective To observe the effects of matrix metalloproteinases (MMP)-2 and MMP-9 secreted by leukemic cells on tight junction proteins ZO-1,claudin-5 and occluding and the permeability of the blood-brain barrier (BBB) and explore the mechanisms of MMP-2 and MMP-9 in leukemic cell infiltration of the central nervous system (CNS).Methods The rnRNA expressions of MMP-2 and MMP-9 in leukemic cell lines SHI-1,HL-60 and U937 were detected by quantitative RT-PCR.The MMP inhibitor GM6001 was used to inhibit the secretion of MMP-2 and MMP-9.RNA interference (RNAi) was used to knock down the expression of MMP-2 and MMP-9.Zymography was used to analyze the secretion of MMP-2 and MMP-9 in the supematant of different leukemia cell lines treated or untreated with drugs,as well as the RNAi-treated cells.An in vitro BBB model composed of human brain microvascular endothelial cells (BMVECs) was developed on a Matrigel-based insert.Cell invasion through a barrier of Matrigelbased human basement membrane and the BMVECs-based human BBB barrier was assayed to measure the invasive capacity and the capacity to breakdown the BBB of different leukemia cell lines treated or untreated with drugs,as well as the RNAi-treated cells.The morphologic changes of BMVECs after co-culture with different leukemia cell lines treated or untreated with drugs,as well as the RNAi-treated cells in vitro BBB models were observed by invert microscopy and tight junction proteins in these BMVECs were analyzed with a laser-scanning confocal microscope.Results (①)The mRNA expression in different leukemic cell lines shown a pronounced transcription of MMP-2 and-9,and the transcriptional level in SHI-1 cells was the highest among all leukemic cell lines tested (P＜0.01).The data of activities of MMP-2 and-9 were consistent with the results of mRNA expression and SHI-l displayed higher capacity of invasion (P＜0.01).(②)After incubation 24h with different leukemic cells,the BMVECs disrupted to loss cell-cell contacts and grew in single cell.Confocal imaging showed down-regulations of ZO-1,claudin-5 and occluding accompanied by the disruption of BBB in vitro models.SHI-1 cells had stronger alterations to BMVECs,tight junction proteins and the permeability of the BBB than HL-60 and U937 cells.However,GM6001 and the knock-down of MMP-2 and MMP-9 altered the responses of BBB.They reduced the degradation of three tight junction proteins with a decreased permeability of BBB.Conclusion MMP-2 and MMP-9 secreted by leukemic cells could disrupt the BBB by degrading the tight junction proteins ZO-1,claudin-5 and occluding,which contributed the infiltration of leukemic cell into CNS.

OBJECTIVETo investigate the effect of up-regulation of Rap1GAP on the invasion ability of leukemic HL-60 cells in vitro, and to establish leukemia mouse model to verify the effects in vivo.

METHODSQuantitative RT-PCR and Western blot methods were used to detect the expression of Rap1GAP in Venus/HL-60 (vehicle control) and Rap1GAP/HL-60 cells (R1 andR2). Transwell method was used to examine the invasion ability in vitro. Quantitative RT-PCR and gelatin zymograph were used to study the expression of MMP-2 and MMP-9. Four-week-old BALB/c nu/nu mice were pre-treated and inoculated with leukemic cells from different groups, several index including survival time were then monitored.

RESULTSRap1GAP mRNA level of R1 and R2 increased about 16-17 folds as compared to the control cells. The invasion rate of R1 and R2 are (55 ± 5)% and (59 ± 4)%, which are significantly higher than (14 ± 4)% of the control cells. The mRNA level of MMP-9 was up-regulated about 12.0 folds in R1 and R2 cells compared to the corresponding control cells. The median survival times of R1 and R2 mice are (32.00 ± 1.85) d and (33.37 ± 2.50) d, respectively, which are shorter than (43.62 ± 2.32) d of the control group. Three mice of R1 and R2 groups showed leukemic cells infiltration in meninges tissue, and the genes of Rap1GAP and MMP-9 were amplified by PCR method.

CONCLUSIONUp-regulated expression of Rap1GAP increased the invasion ability of HL-60 cells accompanied with enhancement of MMP-9 expression in vitro, and the experiment in mouse model also confirmed that Rap1GAP enhanced the invasion of HL-60 cells in vivo.

Animals ; GTPase-Activating Proteins ; metabolism ; HL-60 Cells ; Humans ; Leukemia ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; RNA, Messenger ; Transcriptional Activation ; Up-Regulation

OBJECTIVETo explore the impact of ITD mutation characteristics on the overall survival (OS) and complete remission duration (CRD) in FLT3-ITD positive non-M3 acute myeloid leukemia (AML).

METHODSCapillary electrophoresis was used to detect the FLT3-ITD characteristics after PCR amplication. Single or multiple mutations were identified by the numbers of peak. FLT3-ITD mutation burden was calculated by the peak area of mutant divided by the wild-type and mutant peak areas. Clinical data was collected and followed up in the FLT3-ITD mutation patients.

RESULTSMultiple ITD mutations were common in patients aged 60 and above. Patients with single ITD mutation had higher percentage of blasts in bone marrow than multiple ITD mutations (0.758 vs 0.638, P=0.028). The numbers and length of FLT3-ITD mutation had no impact on prognosis. Patients with less than 10% of ITD mutation burden showed no difference with the intermediate-risk c-kit group in OS and CRD, but the two groups had longer OS and CRD than ITD mutation burden above 10% (OS: undefined, undefined, 9.9 months, P<0.05; CRD: undefined, undefined, 6.7 months, P<0.05). In patients with ITD mutation burden above 10%, cases with NPM1 or CEBPA mutation alone had markedly longer CRD than ITD mutation alone (25.0 vs 5.1 months, P=0.003), while OS were similar (11.4 vs 8.0 months, P>0.05).

CONCLUSIONNon-M3 AML patients with less than 10% FLT3-ITD mutation burden had a better prognosis than those above 10%.

Genotype ; Humans ; Leukemia, Myeloid, Acute ; Mutation ; Prognosis ; Remission Induction ; fms-Like Tyrosine Kinase 3

Bone Marrow ; Hematologic Diseases ; Hematopoietic Stem Cells ; Humans

Adolescent ; Adult ; Female ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid ; genetics ; Male ; Oncogene Proteins, Fusion ; genetics ; Retrospective Studies

BACKGROUNDMMPs and TIMPs play important roles in tumor angiogenesis and invasion. Studies have shown that TIMP-2 has two roles in tumor invasion. However, its role in leukemic infiltration has not been well investigated. This study explored the roles of TIMP-2 in extramedullary infiltration of acute monocytic leukemic SHI-1 cells both in vitro and in vitro.

METHODSA retroviral vector carrying the human TIMP-2 cDNA was constructed and transfected into the monocytic leukemic cell line SHI-1. The expression of TIMP-2 in the positive clones was determined. The proliferation of SHI-1 cells was examined by MTT assay. Trans-Matrigel invasion assays were used to investigate the infiltration ability in vitro. SHI-1 cells were intravenously injected into pre-treated nu/nu mice to investigate the infiltration ability feature in vitro.

RESULTSThe expression of TIMP-2 on the cell membrane was significantly elevated in SHI-1/TIMP-2 cells. Over-expression of TIMP-2 promoted the cells proliferation and the invasions in vitro. The SHI-1/TIMP-2 cells demonstrated higher infiltration ability when intravenously injected into nu/nu mice.

CONCLUSIONOver-expression of TIMP-2, especially on the cell membrane, may play important roles in promoting the proliferation and infiltration of SHI-1 leukemic cells.

Adult ; Animals ; Cell Line ; Cell Proliferation ; physiology ; Humans ; Leukemic Infiltration ; physiopathology ; Male ; Mesenchymal Stromal Cells ; metabolism ; physiology ; Mice ; Mice, Inbred BALB C ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; metabolism

Background MMPs and TIMPs play important roles in tumor angiogenesis and invasion.Studies have shown that TIMP-2 has two roles in tumor invasion.However,its role in leukemic infiltration has not been well investigated.This study explored the roles of TIMP-2 in extramedullary infiltration of acute monocytic leukemic SHI-1 cells both in vitro and in vitro.Methods A retroviral vector carrying the human TIMP-2 cDNA was constructed and transfected into the monocytic leukemic cell line SHI-1.The expression of TIMP-2 in the positive clones was determined.The proliferation of SHI-1 cells was examined by MTT assay.Trans-Matrigel invasion assays were used to investigate the infiltration ability in vitro.SHI-1 cells were intravenously injected into pre-treated nu/nu mice to investigate the infiltration ability feature in vitro.Results The expression of TIMP-2 on the cell membrane was significantly elevated in SHI-1/TIMP-2 cells.Over-expression of TIMP-2 promoted the cells proliferation and the invasions in vitro.The SHI-1/TIMP-2 cells demonstrated higher infiltration ability when intravenously injected into nu/nu mice.Conclusion Over-expression of TIMP-2,especially on the cell membrane,may play important roles in promoting the proliferation and infiltration of SHI-1 leukemic cells.

Objective To investigate the effect of Shanzha Xiaozhi capslue on intima-media thickness(IMT) and plaques score of carotid artery in patients with non-acute phase coronary heart disease and traditional Chinese medicine(TCM)syndrome of phlegm and blood stasis. Methods A prospective study was conducted to carry out a research on 102 patients with non-acute phase coronary heart disease and TCM syndrome of phlegm and blood stasis. They were randomly divided into two groups:a control group(50 cases)treated with conventional western medicine alone and a observation group(52 cases)which was treated by both conventional western medicine and Shanzha Xiaozhi capslue(the main TCM ingredients:Shanzha,Dahuang)0.7 g,3 times a day,the therapeutic course being 6 months in both groups. The cardiocerebral vascular incidences of the two groups were observed,in the mean time,the carotid artery IMT and plagues score were registered,the scores of phlegm stagnation syndrome and blood-stasis syndrome were measured,adverse reaction was observed and compared in both groups before and after treatment. Results After treatment,the occurrence of major adverse acute cardiovascular and cerebrovascular events in observation group were reduced significantly compared with those in the control group(5.77%vs. 20.00%,P<0.05). The IMT and plaques score of carotid artery and the scores of phlegm stagnation syndrome and blood-stasis syndrome were all decreased obviously compared to those before treatment,and the effect was more remarkable in the treatment group〔IMT of carotid artery(mm):0.80±0.13 vs. 0.95±0.12,the plaques score:1.35±0.65 vs. 1.75±0.88, phlegm syndrome score:20.98±6.42 vs. 35.55±9.22,blood-stasis syndrome score:23.23±5.12 vs. 28.95±6.38, P<0.05 or P<0.01〕. Abdominal pain appeared in 2 patients of observation group without other adverse reactions. Conclusion Shanzha Xiaozhi capsule can stabilize atherosclerotic plaque and reduce the occurrence of acute adverse cardiocerebral vascular events in patients with non-acute phase coronary heart disease and phlegm and blood stasis syndrome possibly by removing phlegm and blood stasis to decrease the IMT and plaques score of carotid artery.