Expression and Activity Regulation of Indoleamine 2,3-Dioxygenase in Acute Myeloid Leukemia Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression and activity regulation of indoleamine 2,3-dioxygenase(IDO) in acute myeloid leukemia cells.</p><p><b>METHODS</b>Expression of IDO and TLR9 in HL-60 and K562 cells cocultured with or without IFN-γ,Tα1,IFN-γ+Tα1 and chloroquine were determined by reverse transcription-polymerase chain reaction(RT-PCR). Then, the IDO activity in HL-60 and K562 cells cocultured with or without IFN-γ,Tα1,IFN-γ+ Tα1 was assayed by coomassie brilliant blue staining and modified colorimetric method.</p><p><b>RESULTS</b>Both IDO and TLR9 mRNA were expressed in HL-60 and K562 cells; IFN-γ increased the expression and activity of IDO in a concentration-dependent manner; Tα1 decreased the expression and activity of IDO in a concentration-dependent manner; the up-regulation of IFN-γ on IDO induced expression and activity had been weakened by Tα1(P<0.01); Chloroquine had no effect on the expression of IDO. The expression of TLR9 in HL-60 cells and K562 cells cocultured with IFN-γ,Tα1,IFN-γ+Tα1 and chloroquine was not significantly changed.</p><p><b>CONCLUSION</b>IDO can be expressed in acute myeloid leukemia cells and possesses the activity. IDO may play an important role in immune tolerance induced by leukemia cells, and become a new predictor of AML prognosis. Tα1 decreases the expression and activity of IDO, which can weaken the induction of IFN-γ on IDO expression and activity, thus Tα1 as an immune modulator may be a new agent for AML immunotherapy.</p>

Effect of quercetin on doxorubicin-induced expression of MDR1 gene in HL-60 cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>Leukemia cells can acquire a multidrug resistant (MDR) phenotype in response to a wide variety of chemotherapeutic agents including doxorubicin (Dox). In addition to the constitutive expression in the leukemia prior to chemotherapy, a complex phenotype of pleiotropic resistance is presented in the residual or recurrent leukemia. Recent studies showed Dox-induced coexpression of COX2 and MDR1 genes in human leukaemia cells, and whether Dox-induced MDR1 up-regulation in acute leukaemia cells is dependent on COX2-transcriptional activity and thus might be overcome or prevented with COX2-promotor inhibitor quercetin interfering with COX2 expression and activity. This study was purposed to investigate the impacts of quercetin on Dox-induced mRNA expression of MDR1 and COX2 genes in HL-60 leukemia cells.</p><p><b>METHODS</b>The MDR1 and COX2 mRNA expression in HL-60 cells was detected by RT-PCR; the prostaglandin E2 (PGE2) release was measured by ELISA; the cytotoxicity of Dox was determined by MTT test.</p><p><b>RESULTS</b>The incubation of HL-60 cells with Dox not only up-regulated MDR1 mRNA, but also COX2 mRNA expression, and after co-incubation with quercetin or celecoxib, Dox-induced overexpression of MDR1 and COX2 mRNA were reduced by quercetin, not by celecoxib, whereas PGE2 release was significantly decreased with subsequent enhancement of Dox cytotoxic efficacy by both of them.</p><p><b>CONCLUSIONS</b>Dox-induced MDR1 up-regulation may be dependent on COX2-transcriptional activity, not PGE2, suggesting that the existence of causal link between COX2 and MDR1 expression induced by Dox, and modulation of COX2 transcriptional expression by quercetin would not only sensitize leukemia cells to Dox, but also prevent the acquisition of MDR during chemotherapy.</p>

Characteristics of bone tunnel changes after anterior cruciate ligament reconstruction using Ligament Advanced Reinforcement System artificial ligament / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>There are different materials used for anterior cruciate ligament (ACL) reconstruction. It has been reported that both autologous grafts and allografts used in ACL reconstruction can cause bone tunnel enlargement. This study aimed to observe the characteristics of bone tunnel changes and possible causative factors following ACL reconstruction using Ligament Advanced Reinforcement System (LARS) artificial ligament.</p><p><b>METHODS</b>Forty-three patients underwent ACL reconstruction using LARS artificial ligament and were followed up for 3 years. X-ray and CT examinations were performed at 1, 3, 6, 12, 24, and 36 months after surgery, to measure the width of tibial and femoral tunnels. Knee function was evaluated according to the Lysholm scoring system. The anterior and posterior stability of the knee was measured using the KT-1000 arthrometer.</p><p><b>RESULTS</b>According to the Peyrache grading method, grade 1 femoral bone tunnel enlargement was observed in three cases six months after surgery. No grade 2 or grade 3 bone tunnel enlargement was found. The bone tunnel enlargement in the three cases was close to the articular surface with an average tunnel enlargement of (2.5 ± 0.3) mm. Forty cases were evaluated as grade 0. The average tibial and femoral tunnel enlargements at the last follow-up were (0.8 ± 0.3) and (1.1 ± 0.3) mm, respectively. There was no statistically significant difference in bone tunnel width changes at different time points (P > 0.05). X-ray and CT measurements were consistent.</p><p><b>CONCLUSIONS</b>There was no marked bone tunnel enlargement immediately following ACL reconstruction using LARS artificial ligament. Such enlargement may, however, result from varying grafting factors involving the LARS artificial ligament or from different fixation methods.</p>

An experimental study on the regulation of bone marrow-derived mesenchymal stem cells through indoleamine 2,3-dioxygenase signaling pathway by thymosin α1 for improving the immunosuppression mediated by T cell / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the regulatory effect of thymosin α1 (Tα1) on immunosuppression of bone marrow mesenchymal stem cells (MSCs) from children with aplastic anemia (AA) through Toll-like receptor 9(TLR9)and indoleamine 2,3-dioxygenase (IDO) signaling pathway.</p><p><b>METHOD</b>Bone marrow T cell subsets from children with AA and normal individuals were measured by using flow cytometry. Expressions of TLR9/IDO mRNA of MSCs cocultured with Tα1 were determined by reverse-transcription PCR (RT-PCR). Inhibition of PHA-activated T cell proliferation and activation by MSCs cocultured with Tα1 was detected by using MTT assay and flow cytometry.</p><p><b>RESULT</b>CD4(+)/CD8(+) ratio (0.64 ± 0.02) in children with AA was significantly lower than that in normal individuals (1.42 ± 0.05); but CD8(+)/CD38(+) ratio (0.92 ± 0.04) was significantly higher than that in normal individuals (0.65 ± 0.05). AA MSCs obviously expressed TLR9, but not IDO; AA MSCs treated with Tα1 downregulated TLR9 expression but upregulated IDO expression in concentration- and time-dependent manners. The inhibition of AA MSCs on T cell proliferation (21.38% ± 12.34%) was lower than that in normal individuals (62.72% ± 17.79%, P < 0.05), while AA MSCs treated with Tα1 for 18 h exhibited a stronger inhibition (42.83% ± 16.54%, P < 0.05).</p><p><b>CONCLUSION</b>The immunosuppression mediated by MSCs could be improved by Tα1 through upregulation of IDO expression via TLR9-dependent signaling pathway. This research provides a new idea for targeted immunomodulatory therapy with bone marrow MSCs from children with AA.</p>

Regulatory effect of thymosin α1 on expression of tlr9/ido mRNA in bone marrow mesenchymal stem cells from children with aplastic anemia / 中国实验血液学杂志

The purpose of this study was to explore the regulatory effect of thymosin α1 (Tα1) on expression of TOLL-like receptor 9 (TLR9)/indoleamine2, 3-dioxygenase (ido) mRNA in bone marrow mesenchymal stem cells (MSC) from children with aplastic anemia (AA). Culture system of bone marrow MSC from AA children and normal children in vitro was established, and the effects of Tα1 on expressions of tlr9 mRNA and ido mRNA of MSC from AA children and normal children were determined by RT-PCR. The results showed that the bone marrow MSC from normal children did not express tlr9 and ido mRNA. Bone marrow MSC from children with AA obviously expressed tlr9 mRNA , but did not express ido mRNA; AA children's MSC treated with Tα1 for 18 hours markedly down-regulated tlr9 mRNA expression, but up-regulated ido mRNA expression in the concentration- and time-dependent ways. It is concluded that Tα1 can up-regulate the expression of ido mRNA in bone marrow MSC from children with AA.

Regulation of hypoxia-induced mRNA expressions of HIF-1alpha and osteopontin and in vitro radiosensitization by tirapazamine in human nasopharyngeal carcinoma HNE-1 and CNE-1 cells / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Combined hypoxic cytotoxic drugs and chemoradiotherapy is an important mean of oncotherapy, and Tirapazamine (TPZ) is one of the most remarkable drugs. It has been shown that TPZ has a synergistic effect with radiotherapy on tumor cells, but whether TPZ would down-regulate the expression of the hypoxia-induced genes has not been reported. This study was to investigate the hypoxia-induced mRNA expressions of hypoxia inducible factor-1alpha (HIF-1alpha) and osteopontin (OPN) in human nasopharyngeal carcinoma HNE-1 and CNE-1 cells and the radiosensitization of TPZ, a hypoxia-specific drug, on HNE-1 and CNE-1 cells in vitro.</p><p><b>METHODS</b>The IC50 values of TPZ for HNE-1 and CNE-1 cells were measured using MTT assay, and the mRNA expressions of HIF-1alpha and OPN in HNE-1 and CNE-1 cells was determined using RT-PCR under aerobic and hypoxic conditions, respectively. The survival rates of HNE-1 and CNE-1 cells treated with or without TPZ at IC10 in the presence or absence of oxygen for 6 h were determined using colony formation assay following exposure to 1-6 Gy of 60Co radiation. The dose-survival curves were plotted and the values of D0, Dq and SER were calculated as a single-hit multitarget model.</p><p><b>RESULTS</b>The IC50 values of TPZ were 34.81 μmol/L and 35.02 μmol/L in HNE-1 and CNE-1 cells under aerobic condition, and 30.20 μmol/L and 28.48 μmol/L under hypoxic condition, respectively. The expressions of HIF-1alpha and OPN mRNA were reduced by TPZ in HNE-1 cells, but not in CNE-1 cells under hypoxic condition. For the HNE-1 cells, the respective values of D0 and Dq were 0.89 Gy and 0.28 Gy following normoxic irradiation versus 1.47 Gy and 0.44 Gy following hypoxic irradiation. For the CNE-1 cells, the respective values of D0 and Dq were 0.72 Gy and 0.68 Gy following normoxic irradiation versus 0.95 Gy and 0.56 Gy following hypoxic irradiation. The values of D0 and Dq for HNE-1 and CNE-1 cells treated with TPZ under hypoxic condition following irradiation were 0.66 Gy, 0.21 Gy and 0.85 Gy, 0.79 Gy, respectively.</p><p><b>CONCLUSION</b>TPZ can down-regulate hypoxia-induced expression of HIF-1alpha and OPN mRNA of HNE-1 cells and radiosensitize the HNE-1 cells but not CNE-1 cells, and act as a hypoxia modifier.</p>

Application of modified quadruple stapling technique in radical proximal gastrectomy for gastric carcinoma / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility and safety of modified quadruple stapling technique in radical proximal gastrectomy for gastric carcinoma.</p><p><b>METHODS</b>Medical records of 55 consecutive patients who underwent radical proximal gastrectomy for gastric cancer were analyzed retrospectively. Twenty-eight patients (modified group) undergoing modified quadruple stapling technique were compared to 27 patients (traditional group) who underwent traditional approach during the same period.</p><p><b>RESULTS</b>There was no perioperative mortality. All the patients had negative pathological resection margin. The mean operative time in the modified group was significantly shorter than that in the traditional group [(158±31) min vs. (195±42) min, P<0.05]. There were no immediate complications such as stricture, bleeding or leakage at the anastomosis, gastroparesis, and wound infection. Postoperative recovery did not differ between the two groups (P>0.05). During the follow-up (range: 3 months-2 years), 2 (7.1%) patients in the modified group and 2 (7.4%) in the traditional group developed reflux esophagitis (P>0.05) and anastomotic inflammation occurred in 2 cases (7.1%) for the modified group and 8 (29.6%) for the traditional group (P<0.05).</p><p><b>CONCLUSION</b>Modified quadruple stapling technique is a feasible and safe method in radical proximal gastrectomy.</p>

Cruciate ligament reconstruction using LARS artificial ligament under arthroscopy: 81 cases report / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>There are many different materials used for ligament reconstruction. Currently, autograft, allograft, and artificial ligaments are used in the reconstruction. The objective of this study was to explore the clinical result of cruciate ligament reconstruction under arthroscopy.</p><p><b>METHODS</b>Eighty-one cases were reconstructed with the LARS ligament under arthroscopy, including 43 cases of anterior cruciate ligament (ACL) injury, 20 cases of posterior cruciate ligament (PCL) injury, and 18 cases of ACL combined with PCL injuries of the knee. The follow up period was 10 to 49 months. The International Knee Documentation Committee (IKDC) and Lysholm knee score scales were used for functional evaluation. We examined the anterior and posterior stability of the knee with KT-1000.</p><p><b>RESULTS</b>According to the Lysholm knee function score scale, the average preoperative score of (44.6+/-1.4) increased to a postoperative score of (82.8+/-2.5) in the ACL group and from (46.6+/-2.3) to (80.8+/-2.0) in the PCL group. In the ACL combined with PCL injury group, the preoperative score increased from (45.2+/-1.2) to (85.5+/-2.3). According to IKDC score standards, in ACL group we evaluated 19 cases as C and 24 cases as D, preoperatively, and postoperatively 27 cases as A, 14 cases as B and two cases as C. In the preoperative PCL group, we had 11 cases defined as C and nine cases as D that resolved to 12 cases as A, seven as B and one case of C in postoperative evaluation. In the ACL combined with PCL injury group we defined four cases as C and 14 as D during preoperative scoring. These patients had postoperative grades of six cases as A, 10 as B, and two cases as C. All of the results have statistical significance.</p><p><b>CONCLUSIONS</b>ACL, PCL, or combined ACL and PCL reconstruction using the LARS ligament under arthroscopy is a minimally invasive, safe and effective method to treat cruciate ligament injuries of the knee. Clinical results are satisfactory in the short term.</p>

In vivo study on influence of a discrete nano-hydroxyapatite on leukemia P388 tissue in BALB/C mice / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the influence of a discrete nano-hydroxyapatite crystal (nano-HAp) on lymphatic leukemia P388 behavior by in vivo techniques.</p><p><b>METHODS</b>A nano-HAp was prepared by a neutralization reaction of 0.1 mol calcium hydroxide suspension and 0.06 mol phosphoric acid solutions at room temperature over pH7. The various doses of the nano-HAp only and the nano-HAp mixture with cyclophosphamide (CY) were injected into mice inoculated with solid tumor lymphatic leukemia P388 and dispersed into PRMI 1640 media harvested the leukemia P388 cells. Sixty P388 BALB/C mice were randomly grouped; 36 of them were used as nano-HAp treated groups and 24 mice as the control groups. The leukemia growth in the mice was examined morphologically, histopathologically and under a transmission electron microscope (TEM).</p><p><b>RESULTS</b>The nano-HAp was identified as a hydroxyapatite by an X-ray diffractometry (XRD) and a Fourier transform infrared spectroscopy (FTIR). The morphology and sizes were observed under a TEM. The tissue growth inhibition ratio (weight%) of solid lymphatic leukemia P388 bearing mice treated with nano-HAp at doses 35 mg/kg, 53 mg/kg and nano-HAp (53 mg/kg) combined with CY (35 mg/kg) in 3 consecutive days via intraperitineal injections were 14.95%, 32.67% and 60.45% respectively. Apoptosis of P388 cell cocultured with nano-HAp was confirmed by TEM.</p><p><b>CONCLUSIONS</b>The tissue growth restriction of solid tumor lymphatic leukemia P388 was greater after an injection of nano-HAp only or nano-HAp mixed with CY than that obtained after injection with physiological saline solution as a control (P < 0.01), and the tissue growth restriction of solid tumor after an injection of nano-HAp combined with CY was greater than that obtained after nano-HAp or CY injection only (P < 0.01).</p>

Effect of every-other-day dose Simvastatin in patients with hyperlipidemia / 中国实用内科杂志

Objective To compare the effects of every-other-day dose simvastatin administration with that of daily therapy of same dose.Methods This was a randomized,prospective,nonblinded clinical trial.The 186 patients with high low-density lipoproteim cholesterol(LDL-C) and/or high total cholesterol (TC),triglycerides (TG),low high-density lipoprotein cholesterol(HDL-C)was studied.All patients were randomized into two groups.The every-other-day do- sing group recerived 20mg of simvastatin in alternate-day and daily dosing group received 20mg of simvastatin every day.Before and after 6 weeks,12 weeks of treatment,serum lipoprotein,Live function tests and ereatine kinase con centra- tion and so on were drawn and bad-side effect were studies.Results The two groups significantly reduce LDL-C,TC, TG and a little increased HDL-C compared with baseline.No stalistically significant differences existed between the two groups in percentage in decrease in lipoprotain at 6 weeks,12 weeks compared with baseline.The bad-side effect in the two groups also had not a singnificant different.Conclusion The every-other-day dose of simvastatin have similar effi- cacious and safe to daily dosing in patients with hyperlipidemia and some cost savings.It can take a primary prevention to coronary heart disease in patients with relatively low-risk hyperlipidemia.