Clinical Significance of CRLF2 High Expression in Bone Marrow Mononuclear Cells from Children with Acute Lymphoblastic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To detect the expression of CRLF2 in bone marrow mononuclear cells from children with newly diagnosed acute lymphoblastic leukemia(ALL) and to explore its clinical significance in pediatric ALL.@*METHODS@#A total of 218 children with newly diagnosed ALL who achieveal the complete remission and had the complete follow-up information were selected, and the expression level of CRLF2 in bone marrow mononuclear cells of these children was detected by real-time fluorescent quantitative PCR, and the significance of CRLF2 expression level in clinical prognosis of ALL children was analyzed by using statistical method.@*RESULTS@#28 cases in 218 children with complete data showed high expression of CRLF2. The cumulative recurrence rate in the CRLF2 high expression group was significantly higher than that in the low expression group (53.6% vs 12.6%) (P＜0.01). The predicted 5-year recurrence-free survival rate (RFS) of ALL children with CRLF2 high expression was significantly higher than that of low expression group (P＜0.01). There was no significant difference in the predicted 5-year RFS between ALL children with CRLF2 low and high expression in the standard-risk(SR) group (P＞0.05). The predicted 5-year RFS of ALL children with CRLF2 low expression was higher than that of ALL children with CRLF2 high expression in the intermediate-risk (IR) and high-risk (HR) groups. (P＜0.05). Cox analysis showed that CRLF2 high expression is an independent risk factor for the relapse of children with ALL.@*CONCLUSION@#The recurrence rate of pediatric ALL with CRLF2 high expression is high, and CRLF2 high expression is an important prognostic factor for high risk of relapse in ALL children with IR and HR. It is necessary to use CRLF2 expression as an indicator of risk stratification in pediatric ALL.

Comparison of multi-mode and single-mode intraoperative neurophysiological monitoring in the treatment of severe ossification of the cervical posterior longitudinal ligament with anterior cervical corpectomy with fusion / 中国骨伤

OBJECTIVE@#To compare the application of multi-mode and single-mode intraoperative neurophysiological monitoring in the treatment of severe ossification of posterior longitudinal ligament of cervical spine with anterior cervical corpectomy with fusion.@*METHODS@#From April 2015 to June 2018, 32 patients with severe ossification of the posterior longitudinal ligament were treated in the Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine. There were 21 males and 11 females, aged 45 to 73 years old, with a mean age of 59 years old. The duration of the disease ranged from 6 to 72 months, with a mean of 39 months. The main manifestations were numbness, numbness and weakness of limbs, cotton feeling of foot stepping on lower limbs, instability of standing and walking. With the gradual aggravation of symptoms, quadriplegia, dysfunction of urine and defecation may occur. Patients with ossification of posterior longitudinal ligament of cervical spine were monitored by somatosensory evoked potentials, motor evoked potentials and electromyogram patterns.@*RESULTS@#During the operation, 8 patients had abnormal amplitude of somatosensory evoked potential(SEP); 5 of them had bleeding during anterior cervical decompression procedure and were placed with too much hemostatic cotton, which caused compression of spinal cord and resulted in abnormal SEP waveform. After removal of the hemostatic cotton, SEP waveform returned ot normal; 3 patients had abnormal SEP waveform due to decreased systolic pressure, which was corrected by increased systolic pressure. Twelve patients had abnormal amplitude of motor evoked potential during monitoring, 9 of which were caused by intraoperative mis-contact with nerve root, and turned to normal after timely adjustment of position, 3 of which were caused by intraoperative inhalation of muscle relaxant during surgery. Among 11 patients with abnormal EMG waveform, 9 patients recovered to normal waveform after adjusting operation, 2 patients recovered to normal waveform after short observation, and all patients recovered to normal waveform of motor evoked potential after operation(<0.05). There were 2 cases of cerebrospinal fluid leakage after operation, which healed spontaneously 7 days after operation, and no complications of spinal cord and nerve occurred in all patients after operation.@*CONCLUSIONS@#In anterior cervical corpectomy with fusion operation for the treatment of severe cervical ossification of posterior longitudinal ligament, various modes of intraoperative neurophysiological monitoring can real-time understand spinal cord and nerve function status, significantly reduce the incidence of spinal cord and nerve injury during operation, and effectively improve the safety of operation.

Effects of minimal residual disease level on day 33 of remission induction and IKZF1 genotype on the survival of children with B-lineage acute lymphoblastic leukemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of minimal residual disease (MRD) level on day 33 of remission induction and IKZF1 genotype on the survival of children with B-lineage acute lymphoblastic leukemia (B-ALL).</p><p><b>METHODS</b>A total of 152 children with newly-diagnosed B-ALL who had complete remission after the first cycle of the chemotherapy and had complete follow-up information were enrolled in this study. According to the MRD detection by flow cytometry on day 33 of remission induction, they were divided into three groups: standard-risk (SR) group (MRD <10; n=60), intermediate-risk (IR) group (10≤ MRD <10; n=55), and high-risk (HR) group (MRD ≥10; n=37). Nested RT-PCR was used to determine the IKZF1 genotype of all children before chemotherapy. The effects of MRD level on day 33 of remission induction and IKZF1 genotype on the recurrence-free survival (RFS) of children with B-ALL were analyzed.</p><p><b>RESULTS</b>There were 7 common IKZF1 subtypes in all the 152 children with B-ALL: IK1, IK2/3, IK4, IK6, IK8, IK9, and IK10. Of the 152 children, 130 had functional subtypes of IKZF1 and 22 had non-functional subtypes of IKZF1. During the follow-up period, relapse occurred in 26 (17%) children, and the recurrence rate was highest in the HR group (P<0.05). However, there was no significant difference in the recurrence rate between the SR group and the IR group (P>0.05). The cumulative recurrence rate of the children with non-functional subtypes of IKZF1 was significantly higher than that of those with functional types of IKZF1 (P<0.01). The predicted 5-year RFS rates in the SR, IR, and HR groups were (94.2±2.9)%, (86.7±3.8)%, and (56.2±4.5)% respectively (P<0.05). The 5-year RFS rate of the children with functional subtypes of IKZF1 was significantly higher than that of those with non-functional subtypes of IKZF1 (P<0.01). There was no significant difference in the predicted 5-year RFS rate between the children with functional subtypes of IKZF1 and those with non-functional subtypes of IKZF1 in the SR group (P>0.05). However, the predicted 5-year RFS rate of the children with functional subtypes of IKZF1 was significantly higher than that of those with non-functional subtypes of IKZF1 in the IR group and the HR group (P<0.05).</p><p><b>CONCLUSIONS</b>B-ALL children with non-functional subtypes of IKZF1 have a high recurrence rate, and the recurrence rate will be even higher in B-ALL children with non-functional subtypes of IKZF1 and MRD ≥10 on day 33 of chemotherapy.</p>

Effect of Icaritin on Proliferation and Apoptosis of THP-1 Cell and Its Mechanism / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of icaritin on the proliferation and apoptosis of THP-1 cells and its mechanism.</p><p><b>METHODS</b>After treated with various concentrations of icaritin, cell proliferation was detected by MTS method, and apoptosis was measured with flow cytometry and Hoechst 33258 staining. Expression of BCL-2, BAX and Caspase-3 protein in THP-1 cell was detected by Western blot.</p><p><b>RESULTS</b>After treatment with various concentrations (4-32 µmol/L) of icaritin for 24, 48, 72 h, the inhibition rate of cell growth significantly increased (P<0.05) in time-dose dependent manner(r=0.946); and the apoptotic rate of cells significantly increased (P<0.05) in time-dose dependent manner(r= 0.924). The expression of BCL-2 protein at 48 h decreased significantly in icaritin-treated group, compared with that in control group (P<0.05), while the expression of BAX and Caspase3 protein at 48 h increased significantly in icaritin-treated group, compared with that in control group (P<0.05).</p><p><b>CONCLUSION</b>Icaritin can inhibit proliferation and induce apoptosis of THP-1 in vitro, Icaritin may induce apoptosis in THP-1 cells through the mitochondrial pathway.</p>

Bridging fixation with locking plate for the treatment of subtrochanteric femoral fractures of Seinsheimer type V / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the clinical effect of bridging fixation with locking plate for the Seinsheimer type V subtrochanteric femoral fracture.</p><p><b>METHODS</b>From March 2009 to September 2014,18 cases of Seinsheimer type V subtrochanteric femoral fracture were treated by open reduction and bridging fixation with locking plate through proximal and distal approach including 16 males and 2 females with an average age of 41 years old ranging from 22 to 67 years old. Among them, 12 cases caused by traffic accident, 5 cases by falling, 1 case by heavy aboving. All cases were fresh and closed fractures. Time between injury and operation was from 4 to 9 days with an average of 6.2 days. Of them, 11 cases were fixed with reverse LISS and the other 7 cases were fixed with anatomical locking plates of proximal femur.</p><p><b>RESULTS</b>The mean time of operation was 110 min (ranged from 90 to 155 min). The mean blood loss during operation was 425 ml (ranged from 350 to 650 ml) and 16 cases got blood transfusion which was meanly 300 ml. The mean hospital time was 14 days (ranged from 12 to 18 days). The mean duration of followed up was 11.8 months (ranged from 8 to 22 months). The mean time of bone union was 6.6 months (ranged from 5 to 8 months). There was not any complication such as infection, implant failure, hip varus, external rotation deformity of low limb or fat embolism. The Sanders hip scores were 53.22 ± 6.48, the result was excellent in 12 cases and good in 6 cases at the last follow-up.</p><p><b>CONCLUSION</b>Under the principle of biological osteosynthesis, treatment of Seinsheimer type V subtrochanteric femoral fracture with bridging locking plate fixation has such advantages as high mechanism, less interference of blood supply, stable fixation and little complication. It is a safe and idea way for the treatment of the Seinsheimer type V subtrochanteric femoral fracture.</p>

Case-control study on close reduction combined with minimally invasive percutaneous plate osteosynthesis for the treatment of distal fracture of tibial shaft / 中国骨伤

<p><b>OBJECTIVE</b>To compare the effects of close reduction combined with minimally invasive percutaneous plate osteosynthesis for distal fracture of tibial shaft.</p><p><b>METHODS</b>From March 2009 to May 2013, there were 124 patients (89 males and 35 females, 30 of them were injured in a traffic accident and 94 were falling down in daily life, the ages ranging from 21 to 81 years old) who suffered from distal fracture of tibial shaft. Sixty-six patients (48 males and 18 females, 45 cases of type A, 12 cases of type B and 9 cases of type C) were treated with close manipulative reduction combined with minimally invasive percutaneous plate fixation. After close reduction, a minimal incision was made and a anatomic plate was inserted just along the medial tibia periostea, and then the fracture was fixed without fracture exposure. The other 58 patients (41 males and 17 females, 41 cases of type A, 10 cases of type B and 7 cases of type C) were treated with conventional open reduction and internal fixation. Length of the incision, operating time, early postoperative pain (recorded using the Visual Analog Scale score) and the outcome results (recorded using the evaluation standard of Johner-Wruhs) were compared.</p><p><b>RESULTS</b>The length of incision was meanly (7.34 ± 1.42) cm in MIPPO group and (21.82 ± 2.35) cm in ORIF group; operation time was (44.48 ± 10.00) min in MIPPO group and (59.42 ± 11.84) min in ORIF group. The postoperative radiographs verified good position of all screws and satisfactory bone fracture reduction in both groups. All the patients were followed up, and the duration ranged from 10 to 24 months (mean 15.2 months) in both groups. In MIPPO group,only one patient had delayed union and got union after Chinese herb therapy. The other 65 patients got bony union during 15 to 20 weeks. While in ORIF group, 3 patients suffered from nonunion and received reoperation with bone grafting, and 4 patients got bone infection.</p><p><b>CONCLUSION</b>Treatment of distal fracture of tibial shaft, combined with close reduction and MIPPO technique, has the advantages such as less invasion, less damage of blood supply, simplified procedure of operation and higher union rate, which is an ideal methods and is accordant to the biological demand.</p>

Case-control study on close manipulative reduction combined with minimally invasive percutaneous plate fixation for the treatment of proximal humeral fractures / 中国骨伤

<p><b>OBJECTIVE</b>To compare the clinical effects of close manipulative reduction combined with minimally invasive percutaneous plate fixation(MIPPO) and conventional open reduction and internal fixation (ORIF) for the treatment of proximal humerus fractures.</p><p><b>METHODS</b>From April 2008 to March 2013, among the 75 patients with fractures of proximal humerus, 26 patients were male and 49 patients were female, ranging in age from 22 to 80 years; 18 patients had injuries caused by traffic accident and 57 patients had injuries caused by falling down. According to Neer classification, there were 49 cases of two-part fractures and 26 cases of three-part fractures. All the patients were divided into two groups: MIPPO group and ORIF group. There were 12 males and 21 females in the MIPPO group,including 22 cases of Neer two parts and 11 cases of Neer three parts, who were treated with close manipulative reduction combined with MIPPO. While the other 42 patients were in the ORIF group,including 16 males and 26 females. Among those patients,27 cases belonged to Neer two parts and 15 cases of Neer three parts, who were treated with ORIF. Length of the incision, blood loss, operating time, early postoperative pain(recorded by VAS), neck-shaft angle of proximal humerus and postoperative function of shoulder(recorded by Constant-Murley score, including pain, function, ROM and muscle length) were compared.</p><p><b>RESULTS</b>The mean lengths of incision were (6.74 +/- 0.38) cm in MIPPO group and (16.82 +/- 1.74) cm in ORIF group;blood losses were (110.15 +/- 29.49) ml in MIPPO group and (326.19 +/- 59.71) ml in ORIF group; operation times were (48.60 +/- 10.18) min in MIPPO group and (68.84-16.22) min in ORIF group. VAS of patients in MIPPO group on the 1st and 3rd days postoperatively were lower than those of patients in the ORIF group. The postoperative radiographs verified good position of all screws and satisfactory reduction of bone fracture reduction in both groups. All the patients were followed up,and the durig ranged from 8 to 24 months (mean 14.2 months). In the MIPPO group, there was no humeral head necrosis and all patients gained bone union; while in the ORIF group, 3 patients sustained nonunion and received reoperation for bone grafting, and 2 patients sustained humeral head necrosis. The mean Constant-Murley scores of shoulder were 88.94 +/- 2.57 in the MIPPO group and 86.00 +/- 3.36 in the ORIF group.</p><p><b>CONCLUSION</b>The close manipulative reduction combined with MIPPO is a better choice for fixation of proximal humerus fractures, compared with conventional plate. This method possesses such advantages as a shorter incision, less disturbance of the blood supply and stable fixation of the fracture, allowing early exercise so that the function of shoulder recovers rapidly.</p>

Close reduction combined with minimally invasive percutaneous plate osteosynthesis for proximal and distal tibial fractures: a report of 56 patients / 中国骨伤

<p><b>OBJECTIVE</b>To evaluate the clinical effects of close reduction combined with minimally invasive percutanous plate osteosynthesis (MIPPO) for proximal and distal tibial fractures.</p><p><b>METHODS</b>From March 2007 to December 2010, 56 patients with proximal and distal tibial fractures were treated with close reduction combined with MIPPO technique. There were 39 males and 17 females,aged from 22 to 67 years with an average of 41.3 years. Left fracture was in 25 cases and right fracture was in 31 cases; proximal tibial fracture was in 15 cases and distal tibial fractures was in 41 cases; 34 cases caused by fall down and 22 cases caused by road accident. The mean time from injury to operation was 1.7 d. Clinical manifestation included pain, swelling of leg with limitation of activity. According to the standard of Johner-Wruhs, clinical effects were evaluated.</p><p><b>RESULTS</b>The mean operative time was 46 min in 56 patients. All fractures obtained satisfactory reduction and the location of plate was good. Incisions healed with one-stage and no superficial or deep infection was found. All the patients were followed up from 8 to 23 months with an average of 14.2 months. Only one fracture complication with delayed union,and after auto grafting with ilium bone,the fracture got union. Other 55 cases obtained bone healing in 15 to 20 weeks after operation and no internal fixation failure was found. The time of walking was 4-6 months after operation,without limping at 7 months after operation. Both lower extremities were symmetrical and the function of knee and ankle got complete recovery. According to the criteria of Johner-Wruhs score,46 cases obtained excellent results,9 good and 2 fair.</p><p><b>CONCLUSION</b>Treatment of proximal and distal tibial fractures with close reduction and MIPPO technique can not only preserve soft tissue,simplify operative procedure and decrease wound, but also can obtain rigid internal fixation and guarantee early function exercises of knee and ankle joints. The method has the advantages of less soft tissue injury, less blood loss, reliable fixation, which is effective method in treating proximal and distal tibial fractures and corresponds with the standpoint of biological fixation.</p>

Effect of glucocorticoid on dendritic cells in children with chronic immune thrombocytopenia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the change in dendritic cells (DCs) in children with chronic immune thrombocytopenia (cITP) and the effect of glucocorticoid on DCs in children with cITP.</p><p><b>METHODS</b>Fifteen children with cITP and 20 healthy controls were included in the study. Flow cytometry was used to measure the DC subsets count in the 15 children with cITP before and after glucocorticoid treatment as well as the corresponding values in the 20 healthy controls. The DCs derived from peripheral blood monocytes in children with cITP were cultured in vitro and collected, and their immunophenotypes were determined by flow cytometry.</p><p><b>RESULTS</b>Before glucocorticoid treatment, the children with cITP showed no notable change in the absolute count of myeloid DCs (mDCs) but showed decreased absolute count of plasmacytoid DCs (pDCs) and increased mDC/pDC ratio compared with the healthy controls (P<0.05). After glucocorticoid treatment, the children with cITP demonstrated increased absolute count of pDCs and decreased absolute count of mDCs and mDC/pDC ratio compared with before treatment (P<0.05). Before glucocorticoid treatment, the children with cITP had significantly higher positive rates of HLA-DR, CD80, CD83 and CD86 on peripheral blood DCs than the healthy controls (P<0.01). All the positive rates were significantly decreased after glucocorticoid treatment (P<0.01), so that there was no significant difference from the healthy controls (P>0.05).</p><p><b>CONCLUSIONS</b>Disproportion and functional disturbance of DC subsets is associated with the pathogenesis of cITP in children. Glucocorticoid can strengthen the immunosuppression of DCs in children with cITP, which may contribute to the effectiveness of glucocorticoid as a treatment.</p>

Influence of the unevenness of articular surface on the osteochondral repair / 中国骨伤

<p><b>OBJECTIVE</b>To investigate whether the unevenness of articular surface would affect the osteochondral repair.</p><p><b>METHODS</b>Eight Shanghai Chongming 6-months-old masculine goats with a mean weight of 25 kg were used in this study. Different unevenness, which were 0.5 mm, 1.0 mm, 2.0 mm protrude or concavity, were created on the weight-bearing portion of the medial femoral condyles of the goats. The goats were sacrificed 12 weeks later and were observed with the general observation, HE staining and transmission electron microscope. To evaluate the microscopic morphology, a histological grading scale described by O'Driscoll, Keeley and Salter was used.</p><p><b>RESULTS</b>The general observation and HE staining showed that the unevenness of 0.5 mm or 1.0 mm protrudes or concavity could be repaired to get the surface smooth on the whole. The transmission electron microscope showed that the reparative tissues were the same as the normal cartilage. The 2.0 mm depth couldn't be repaired satisfactorily. The transmission electron microscope showed that the fiber bundle proliferated and the chondrocytes degenerated. The scores of the 2.0 mm depth were significantly lower than that of the 0.5 mm or 1.0 mm (P < 0.05).</p><p><b>CONCLUSION</b>The unevenness could have an influence on the repair. The limited unevenness could be repaired by itself.</p>

Research on the repair of acute large osteochondral defects with mosaicplasty associated with genes-enhanced tissue engineering in different proportion / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the optimum proportion of Mosaicplasty and genes-enhanced tissue engineering for the repair of acute osteochondral defects.</p><p><b>METHODS</b>Western blot test was conducted to detect the expression of hTGF-beta1, Col II and Aggrecan in 3 groups, including hTGF-beta1, transduction group, Adv-betagal transduction group and control group without transduction. Eighteen 6-month-old Shanghai male goats (weight: 22 to 25 kg) were used. BMSCs were isolated from the autologous bone marrow, and were subcultured to get the cells at passage 3. Thirty-six medial femoral condyles were used and divided into 6 groups named AR, AL, BR, BL, CR, and CL. Acute cylindrical defects (9 mm in diameter and 3 mm in depth)were created in the weight bearing area of the medial femoral condyle of hind limbs. In the single group, the autologous osteochondral mosaicplasty was performed to repair the defect; in the combination group, besides the mosaicplasty, the dead space between the cylindrical grafts and the host cartilage were injected with the suspension of hTGF-beta1, gene enhanced autogenous BMSCs in sodium alginate, and CaCl2 was dropped into it to form calcium alginate gels. The autologous osteochondral transplantation cover rates of group AR was 44.44% single group, AL was 44.44% combination group, BR was 33.33% single group, BL was 33.33% combination group, CR was 22.22% single group, and CL was 22.22% combination group. The goats were killed 24 weeks after operation to receive gross and histology observation, which was evaluated by the histological grading scale of O'Driscoll, Keeley and Salter. Immunohistochemistry and TEM observation were also performed.</p><p><b>RESULTS</b>Western blot test showed the expression of the hTGF-beta1, Col II and the Aggrecan in the hTGF-beta1 transduction group were significantly higher than that of the Adv-betaga1 transduction and the blank control groups. The gross and histology observation revealed that each defects of six groups had different degrees of repairing. There was no significantly difference among the BL, AR, and AL groups. But the scores of the other three groups (BR, CR, and CL) were significantly poorer than the former three groups.</p><p><b>CONCLUSION</b>Mosaicplasty associated with genes enhanced tissue engineering could repair the osteochondral defects effectively. With the autologous osteochondral transplantation coverage reducing, the advantage of the combination could have a better representation.</p>

A study of proximal humerus fractures using close reduction and percutaneous minimally invasive fixation / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the clinical effects of close reduction and percutaneous minimally invasive fixation in the treatment of proximal humerus fractures.</p><p><b>METHODS</b>From April 2008 to March 2010, 28 patients with proximal humerus fracture were treated with close reduction and percutaneous minimally invasive fixation. There were 21 males and 7 females, ranging in age from 22 to 78 years,with an average of 42.6 years. The mean time from suffering injuries to the operation was 1.7 d. Nineteen cases caused by falling down, 9 cases by traffic accident. The main clinical manifestation was swelling, pain and limited mobility of shoulders. According to Neer classification, two part fractures were in 17 cases and three part fractures in 11 cases. The locking proximal humerus plate was used to minimally fixation through deltoid muscle under acromion. The operating time,volume of blood loss, the length of incision and Constant-Murley assessment were applied to evaluate the therapeutic effects.</p><p><b>RESULTS</b>The mean operating time was 40 min, the mean blood loss was 110 ml, and the mean length of incision was about 5.6 cm. The postoperative X-ray showed excellent reduction and the plate and screws were successfully place. Twenty-eight patients were followed up for 6 to 24 months (averaged 14.2 months). The healing time ranged from 6 to 8 weeks and all incision was primarily healed. There were no cases with necrosis head humerus, 24 cases without omalgia, and 4 cases with o-malgia occasionally. All the patients can complete the daily life. The mean score of Constant-Murley assessment was 91.0 +/- 5.8, 24 cases got an excellent result, 3 good and 1 fair.</p><p><b>CONCLUSION</b>Close reduction and percutaneous minimally invasive fixation, not only can reduce surgical invasive, but also guarantee the early function activities. It has the advantages of less invasive, fixed well and less damage of blood circulation.</p>

Effects of PPAR-gamma agonist rosiglitazone on MMP-9 and TIMP-1 expression of monocyte-derived macrophages isolated from patients with acute coronary syndrome / 中华心血管病杂志

<p><b>OBJECTIVE</b>Coronary arterial plaque rupture and secondary thrombosis are the major pathogenesis of acute coronary syndrome (ACS). Metalloprotease (MMPs) secreted by monocyte/macrophage was the main predisposing factor of the plaque rupture and peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is involved in a variety of inflammatory cytokine gene transcriptional regulations. We explored the possible role of PPAR-gamma in the regulation of MMP-9 and TIMP-1 expressed by peripheral monocyte-derived macrophages (MDMs) from patients with ACS.</p><p><b>METHODS</b>Peripheral blood mononuclear cells were isolated from 48 patients with ACS and 28 healthy controls and stimulated by macrophage colony-stimulating factor (0.1 microg/ml for 24 hours) to form MDMs. MDMs were then incubated under various concentrations of rosiglitazone (0, 1, 10, 20 micromol/L) for 48 hours. The concentrations of MMP-9 and TIMP-1 in the supernatant were measured by enzyme linked immunosorbent assay, and the mRNA expression of PPAR-gamma, MMP-9 by RT-PCR and nuclear factor-kappaB P65 (NF-kappaB P65) expression by immunohistochemistry.</p><p><b>RESULTS</b>PPAR-gamma mRNA expression was significantly lower while NF-kappaB P65 and MMP-9 expression as well as MMP-9 and TIMP-1 concentrations in supernatant were significantly higher in ACS group than those in control group (all P < 0.05). After rosiglitazone intervention, PPAR-gamma mRNA expression was significantly upregulated in both ACS and control groups in a dose-dependent manner. Both the MMP-9 concentration in the supernatant and MMP-9 mRNA expression were reduced post intervention with rosiglitazone in both groups. The TIMP-1 mRNA expression and concentration in supernatant were not affected by rosiglitazone in both groups. Rosiglitazone induced significant downregulation of NF-kappaB P65 expression in both groups.</p><p><b>CONCLUSION</b>Rosiglitazone intervention may downregulate MMP-9 expression by upregulating PPAR-gamma expression, and by downregulating NF-kappaB expression in MDMs isolated from patients with ACS.</p>

Case-control studies on debridement combined with external application of Mangxiao for the treatment of severe open injuries of limbs / 中国骨伤

<p><b>OBJECTIVE</b>To investigate simple and convenient methods for the treatment of severe open injuries of limbs.</p><p><b>METHODS</b>From 2005.1 to 2008.3, 31 patients got limb salvage in 33 cases (Gustilo III B) from emergency, which were divided into two groups(A and B). The 16 patients in Group A (14 male and 2 female) were treated with emergency debridement, sequential dressing change, and finally with skin grafting or free flap. The 15 patients in Group B (11 male and 4 female) were treated with radical debridement (skin and muscle), decompression of fascial compartment routinely and repair of periosteal avulsion lesion. The patients in Group B were also treated with external application of Mangxiao postoperatively and dressing change for first time one week later. The clinical effects were assessed by the skin survival rate of degloving injury, the number of operative procedures (using free flap or not), the number of dressing changes in the first two weeks, body temperature and white cell count (48 h after the first debridement ). Statistical analysis was done with SPSS software. Chi-square analyses was used to compare the enumeration data, and compare means with independent samples T test for measurement data.</p><p><b>RESULTS</b>All the patients in Group A got limb salvage (10 patients were upper limb, 6 patients were lower limb), and the mean skin survival rate of degloving injury was (77.88 +/- 4.21)%. Five patients in Group A were finally treated with free flap. All the patients in Group B got limb salvage (10 patients were upper limb, 5 patients were lower limb) by a few secondary skin grafting, and the mean skin survival rate of degloving injury was (97.53 +/- 3.09)%. The mean skin survival rate of Group B was obviously better than that of Group A (P<0.01). The number of dressing changes in the first two weeks in Group B was absolutely less than that of Group A (P<0.01). Also, infection rate and the number of operative procedures of Group B was significantly less than that of Group A (P<0.01). All the patients were followed up for at least 3 months.</p><p><b>CONCLUSION</b>Radical debridement combined with external application of Mangxiao for the treatment of severe open fractures is simple, convenient and effective. It is better for a lot of people injured simultaneously in accident.</p>

Expression characteristics of heat shock protein 70 and pim-1 gene in bone marrow mononuclear cells from leukemia patients and their clinical significance / 中国实验血液学杂志

This study was aimed to investigate the expression characteristics of HSP70 protein/mRNA, pim-1 mRNA in bone marrow mononuclear cells from leukemia patients, and to clarify whether these changes are related to leukemia type, tumor burden of leukemia, therapeutic reaction and prognosis. HSP70 mRNA and pim-1 mRNA in BMMNCs were detected with semi-quantitative RT-PCR in 40 leukemia patients and 10 controls. HSP70 protein in BMMNCs was assayed with Western blot in 34 leukemia patients and 10 controls. Relation of HSP70 and pim-1 expression with leukemia classification, the degree of tumor burden and therapeutic reaction were analyzed. The results showed that the BMMNCs from both leukemia patients and controls expressed HSP70 protein/mRNA. The mean ODR value of HSP70 mRNA in BMMNCs from leukemia patients was significantly higher than that of the controls; the mean ODR value of HSP70 protein/mRNA in acute myeloid leukemia and chronic myeloid leukemia patients both were significantly higher than that of acute lymphocytic leukemia patients; the mean ODR value of HSP70 protein/mRNA in acute leukemia patients with high-degree tumor burden was higher than that of the patients with low-degree tumor burden; the mean ODR value of HSP70 protein/mRNA in the patients after chemotherapy was significantly higher than that of the patients before chemotherapy; the BMMNCs from both leukemia patients and controls expressed pim-1 mRNA. The mean ODR value of pim-1 mRNA in BMMNCs from leukemia patients was significantly higher than that of the controls; the mean ODR value of pim-1 mRNA in BMMNCs for Acute lymphocytic leukemia patients was significantly higher than that of the patients suffered from acute myeloid leukemia and chronic granulocytic leukemia; there was a positive relationship between pim-1 mRNA and HSP70 mRNA expressions in leukemia patients (p < 0.05). It is concluded that there are high expressions of HSP70 and pim-1 in leukemia and their positive correlation is shown. The over-expressions of HSP70 and pim-1 protein/mRNA are related to tumor burden in leukemia patients.

In vitro expansion of the adult human bone marrow mesenchymal stem cells for clinic application in HSCT / 中国实验血液学杂志

This study was aimed to investigate the efficiency of 4 different culture media for in vitro culture and expanding adult human bone marrow mesenchymal stem cells (ahBM-MSCs) so as to establish a protocol of culturing and expanding hBM-MSCs and provide exprimental basis for hematopoietic blood stem cell transplantation combined with BM-MSCs. BM-MSCs were obtained from 16 fresh adult human bone marrow aspirate by gradient centrifugation with Ficoll Paque, then cultured in DMEM/F12 with 10% umbilical cord blood serum, 10% fetal calf serum (FCS), human blood serum, and MesenCult culture medium. The surface antigens of BM-MSCs were detected by flow cytometry. BM-MSCs were differentiated into osteoblasts and adipocytes under culture in the conditioned medium special for osteogenesis, and adipogenesis and the differentiated MSCs were identified by morphological observation, immunophenotype and immunohistochemical staining. The results showed that BM-MSCs could be isolated from adult human bone marrow and cultured by all culture media. The effect of umbilical cord blood serum on BM-MSC proliferation and their purity were similar to that of MesenCult culture medium, but better than that of FCS and human blood serum. The positive rate of CD29, CD73, CD105 on BM-MSCs cultured in umbilical cord serum and MesenCult medium was higher than that in FCS and adult human serum (p < 0.05), and the positive rate of CD31 was lower than that in FCS and adult human serum (p < 0.05). The positive rate of BM-MSCs differentiated into osteoblasts and adipocytes under culture in the conditioned medium for osteogenesis and adipogenesis with umbilical cord blood serum and MesenCult culture medium was also higher than that in FCS and adult human serum (p < 0.05). It is concluded that BM-MSCs can be obtained by all the four methods. DMEM/F12 with 10% umbilical cord blood serum and MesenCult culture medium are better than the others for the purification and differentiation potency of BM-MSCs in vitro. The medium with umbilical cord serum is valuable for clinical application in HSCT.

Effect of the microtubule depolymerization on mitochondria damage in rat myocardiocytes early after hypoxia / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the effect of microtubule depolymerization on mitochondria damage in rat myocardiocytes early after hypoxia.</p><p><b>METHODS</b>Myocardiocytes from Wistar rats were isolated according to routine procedure, and they were randomly divided into control group, depolymerization group (with treatment of 4 micromol/L colchicines in the culture medium), hypoxia group, hypoxia and depolymerization group (with treatment of 4 micromol/L colchicines in the culture medium combined with low oxygen tension). The changes in distribution of the mitochondria were examined with laser confocal microscopy, the morphology and the structure of mitochondria was observed by transmission electron microscope, the respiration control ratio (RCR) was determined by respirometer, and the content of adenosine triphosphate (ATP) in endochylema was detected with liquid chromatograph at 20, 30, 60 post-hypoxia minutes (PHM).</p><p><b>RESULTS</b>In control group, the mitochondria was in granular form, with regular arrangement, while mild changes were observed in depolymerization group. At 20, 30, and 60 PHM, the disarrangement in distribution and morphologic damage were aggravated in hypoxia depolymerization group, and the RCR (1.58 +/- 0.37, 1.51 +/- 0.32, 1.12 +/- 0.11, respectively) were evidently lower than those in hypoxia group (3.85 +/- 0.56, 2.98 +/- 0.44, 1.79 +/- 0.73, respectively, P < 0.01). The content of ATP showed the same tendency at the same time-points (419 +/- 83, 326 +/- 73, 295 +/- 58 ng/mg) compared with hypoxia depolymerization group [(475 +/- 68, 397 +/- 59, 336 +/- 67 ng/mg) in hypoxia group].</p><p><b>CONCLUSION</b>The disarrangement in distribution of mitochondria, as well as the damage in mitochondrial structure, respiratory, function and energy metabolism, can be aggravated by microtubule depolymerization after hypoxia, which indicates that microtubule depolymerization plays an important role in the mitochondria damage.</p>

Study on injury to microtubule of cardiomyocytes at early post-hypoxia stage / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the degree of injury to microtubule of myocardium at early post-hypoxia stage.</p><p><b>METHODS</b>Cardiomyocytes from Wistar rats were isolated and cultured, and they were then divided into normal control and hypoxia groups. The distribution and morphological changes in microtubules were observed with laser confocal microscopy and scanning electron microscope at 10, 20, 30 post-hypoxia minutes (PHM) and 1 post-hypoxia hour (PHH). Then the fluorescence intensity of alpha-microtubule was detected with RT-PCR, the morphology of microtubule was observed, and the expression of dissociative alpha-microtubule was determined by Western blot.</p><p><b>RESULTS</b>Compared with normal control group, the bead-like structure of the microtubule in hypoxia group disappeared at 10 PHM, but no obvious change was observed in the distribution and number of microtubules. Despite the disappearance of bead-like structure of the microtubule, the microtubule derangement and loss of microtubule at the edge of cell were observed at 20 PHM. The fragmentation, derangement of texture, and loss of regularity in cardiomyocytes were observed at 30 PHM and 1 PHH. The fluorescence intensity of alpha-microtubule in hypoxia group was evidently decreased than that in normal group in a time-dependent manner. The expression of dissociative alpha-microtubule in hypoxia group at 10 PHM (46,644 +/- 145) was obviously higher than that in normal group (13,357 +/- 98, P < 0.01), and its increase was maintained with elapse of time.</p><p><b>CONCLUSION</b>Microtubule injury to cardiomyocytes occurs at early stage of post-hypoxia, with destruction of its structure and distribution.</p>

Study on the influence of hypoxia induced microtubule damage on the opening of mitochondrial permeable transition pore of cardiac myocytes in rat / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the influence of hypoxia induced microtubule damage on the opening of mitochondrial permeable transition pore (MPTP)of cardiac myocytes and on the decrease of respiratory function in rat.</p><p><b>METHODS</b>Primary cultured myocardial cells from 30 neonatal rats were randomized as normoxic group (A), hypoxia group (B), normoxia with microtubule destabilizing agent group (C, with treatment of 8 micromol/L colchicines for 30 minutes before normoxia), and hypoxia with microtubule stabilizing agent group (D, with treatment of 10 micromol/L taxol for 30 minutes before hypoxia). beta-tubulin immunofluorescence ,the opening of mitochondria permeability transition pore, and the mitochondrial inner membrane potential were detected at 0.5, 1, 3, 6 and 12 post-treatment hours (PTH), and the mitochondrial respiratory function was determined by MTT method. The changes in these indices were also determined in A group at the corresponding time-points.</p><p><b>RESULTS</b>Obvious damage of polymerized microtubule, opening of MPTP, mitochondrial inner membrane potential loss and decrease of myocardial respiratory activity were observed in both group B and C at 0.5 PTH, and they became more and more serious afterwards. However, the changes in the above indices in D group were much better than those in B group (P < 0.05 or 0.01), and no difference was found between D (92.8 +/- 4.0)% and C [(100.0 +/- 0.0) %, P > 0.05] groups.</p><p><b>CONCLUSION</b>Hypoxia played a role in the myocardial microtubule damage as well as in the opening of MPTP. Moreover, hypoxia could also impair the mitochondrial respiratory function. Microtubule destabilizing agent could reproduce well the process of hypoxia induced microtubule damage, while the stabilizing agent exerted protective effect by improving the transition of mitochondrial permeability and the mitochondria respiratory function.</p>

Correlation between microtubule damage and mitochondria damage in hypoxic cardiomyocytes / 第三军医大学学报

Objective To investigate the relationship between microtubule damage and mitochondria damage in hypoxic cardiomyocytes.Methods Cardiomyocytes from neonatal Wistar rat were isolated and cultured for 3-5 d,then divided into three groups: normal control,hypoxia(cultured in hermetic container containing 1%O_(2),5%CO_(2),95%N_(2)),hypoxia with microtubule depolymerization(by adding 4 ?mol/L colchicine).The fluorescence intensity of ?-microtubule was determined by flow cytometry,the morph and distribution of ?-microtubule and mitochondria were checked by confocal laser scanning microscopy(CLSM) at 30 min,1 h after the cadiomyocytes being oxygen deficit.Results As compared with the normal control cells,the fluorescence intensity of ?-microtubule decreased and the microtubule ruptured and distributed confusedly,and the mitochondria changed into disorder in hypoxic cells.As compared with the hypoxic cells,the distribution of mitochondria confused and the fluorescence intensity decreased in the cells of hypoxia with microtubule depolymerization.Conclusion Microtubule damage occurred at the early stage of hypoxia(30 min).Microtubule damage may lead to mitochondria damage in hypoxic cardiomyocytes.