Effects of Rehmanniae Radix and Rehmanniae Radix Praeparata on proteomics and autophagy in mice with type 2 diabetes mellitus induced by high-fat diet coupled with streptozotocin / 中国中药杂志

To compare the pancreatic proteomics and autophagy between Rehmanniae Radix-and Rehmanniae Radix Praeparata-treated mice with type 2 diabetes mellitus(T2DM). The T2DM mouse model was established by high-fat diet coupled with streptozotocin(STZ, intraperitoneal injection, 100 mg·kg~(-1), once a day for three consecutive days). The mice were then randomly assigned into a control group, low-(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix groups, low-(150 mg·kg~(-1)) and high-dose(300 mg·kg~(-1)) catalpol groups, low-(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix Praeparata groups, low-(150 mg·kg~(-1)) and high-dose(300 mg·kg~(-1)) 5-hydroxymethyl furfuraldehyde(5-HMF) groups, and a metformin(250 mg·kg~(-1)) group. In addition, a normal group was also set and each group included 8 mice. The pancreas was collected after four weeks of administration and proteomics tools were employed to study the effects of Rehmanniae Radix and Rehmanniae Radix Praeparata on protein expression in the pancreas of T2DM mice. The expression levels of proteins involved in autophagy, inflammation, and oxidative stress response in the pancreatic tissues of T2DM mice were determined by western blotting, immunohistochemical assay, and transmission electron microscopy. The results showed that the differential proteins between the model group and Rehmanniae Radix/Rehmanniae Radix Prae-parata group were enriched in 7 KEGG pathways, such as autophagy-animal, which indicated that the 7 pathways may be associated with T2DM. Compared with the control group, drug administration significantly up-regulated the expression levels of beclin1 and phosphorylated mammalian target of rapamycin(p-mTOR)/mTOR and down-regulated those of the inflammation indicators, Toll-like receptor-4(TLR4) and Nod-like receptor protein 3(NLRP3), in the pancreas of T2DM mice, and Rehmanniae Radix showed better performance. In addition, the expression levels of inducible nitric oxide synthase(iNOS), nuclear factor erythroid 2-related factor 2(Nrf2), and heine oxygenase-1(HO-1) in the pancreas of T2DM mice were down-regulated after drug administration, and Rehmanniae Radix Praeparata demonstrated better performance. The results indicate that both Rehmanniae Radix and Rehmanniae Radix Praeparata can alleviate the inflammatory symptoms, reduce oxidative stress response, and increase the autophagy level in the pancreas of T2DM mice, while they exert the effect on different autophagy pathways.

Role of Rheb in Human Acute Myeloid Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the role of Rheb (mTOR activator) in AML development by measuring Rheb expression in bone marrow of adult AML patients and in AML cell line HL-60.</p><p><b>METHODS</b>Real-time PCR assay was used to measure the Rheb mRNA expression in 27 AML patients and 29 ITP patients as control. The relationship between Rheb mRNA expression and age, AML subtype, fusion gene, splenomegaly, hepatomegaly and survival of AML patients was analyzed and compared. In addition, HL-60 cell line over-expressing Rheb was established, and the HL-60 cells and HL-60 cells with overexpression of Rheb were treated with Ara-C of different concentrations, the proliferation level was detected by CCK-8 method, and the IC50 was calculated.</p><p><b>RESULTS</b>The mRNA level of Rheb in AML patients was similar to that in ITP patients (control). Interestingly, higher expression of Rheb was associated with better survival and was sensitive to Ara-C treatment. However, the expression level of Rheb was not associated with age, AML subtype, fusion gene, and hepatomegaly of patients. Lower expression level of Rheb was associated with splenomegaly. In vitro analysis of HL-60 line indicated that overexpression of Rheb could increased the cell sensitivity to Ara-C treatment (IC50=0.54 µmol/L) and caused HL-60 cell apoptosis.</p><p><b>CONCLUSION</b>The lower Rheb expression is a poor prognostic indicator for AML patients, which is associated with AML splenomegaly, the patients and HL-60 cells with low expression of Rheb are insensitive to Ara-C treatment.</p>

Effect of TEB-415, a Derivative of Imatinib, on Multiple Myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the growth inhibitory effect of Imatinib derivative TEB-415 on various multiple myeloma (MM) cell lines, such as U226, H929, RPMI8226, MM1R and MM1S.</p><p><b>METHODS</b>TEB-415, a derivative of Imatinib was synthesized by modifying the chemical structure of Imatinib. MM cell lines (U226, H929, RPMI8226, MM1R and MM1S) were treated with TEB-415, Imatini and Bortezomib of various concentrations. Cells were grown for 72 hours and the growth rate was measured by CCK-8 method, cell morphology was observed and the IC50 was calculated.</p><p><b>RESULTS</b>TEB-415 could inhibit H929 and RPMI8226 growth significantly. When the concentration of TEB-415 was <0.1 nmol/L, >50% H929 cells died. The IC50 of Imatinib was 0.123 mol/L while the IC50 of Bortezomib was 0.03 nmol/L. In RPMI8226 cell line, when the concentration of TEB-415 was 11.9 mol/L, more than 50% of cells died. In contrast, when RPMI8266 were treated with Imatinib of the concentration of 12.8 mol/L, cells grew normally.</p><p><b>CONCLUSION</b>In comparison to Imatinib, TEB-415, a derivative of Imatinib, can kill H929 MM cells much effectively, its effecacy is only inferior to Bortezomib. RPMI8226, an MM cell line is insensitive to Imatinib, but still sensitive to TEB-415 and its growth can be inhibited by TEB-415.</p>

Modified total hepatic vascular exclusion for liver extracapsular resection of giant hepatic cavernous hemangioma / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the feasibility of intraoperative autologous transfusion in modified total hepatic vascular exclusion under normal temperature for extracapsular resection of giant hepatic hemangioma.</p><p><b>METHODS</b>The clinical data of 32 patients undergoing hepatic resection with total hepatic vascular exclusion requiring intraoperative autologous transfusion were analyzed retrospectively. The tumors in these cases involved the proximal hepatic veins and inferior vena cava, with hemangioma volume ranging from 12 cm x 15 cm to 18 cm x 40 cm.</p><p><b>RESULTS</b>The hemangioma were completely resected in all patients, who all recovered smoothly. In one case, hemangioma rupture occurred during dissociation of the liver, resulting in massive hemorrhage which required blood transfusion of 6000 ml. Four patients received blood transfusion of 400-800 ml, and the other 27 had no blood transfusion. Only 8 patients underwent pringle maneuver with resection, whereas the other 27 underwent total hepatic vascular exclusion during liver resection for 5-30 min (mean 16 min).</p><p><b>CONCLUSION</b>Intraoperative autologous transfusion in modified total hepatic vascular exclusion under normal temperature is feasible and safe for extracapsular resection of huge hepatic hemangioma adjacent to the major arteries.</p>

Migration of 99mTc-Hexamethylpropylene Amine Oxime (HMPAO) Labeled Immature and Mature Dendritic Cells in the Mouse / 대한핵의학회잡지

PURPOSE: The purpose of this study is to evaluate migration of technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO) labeled immature and mature dendritic cells (DC) in the mouse. METHODS: DC were collected from bone marrow (BM) of tibiae and femurs of mice. Immature and mature DC from BM cells were radiolabeled with 99mTc-HMPAO. To evaluate the functional and phenotypic changes of DC from radiolabeling, the allogeneic mixed lymphocyte reaction (MLR) and fluorescence-activated cell sorting (FACS) analysis were performed before and after labeling with 99mTc-HMPAO. Migration of intravenously injected DC (iv-DC) was assessed by serial gamma camera images of mice with or without subcutaneous tumor. Percent injected dose per gram (%ID/g) was calculated in lungs, liver, spleen, kidneys, and tumor through dissection of each mice after 24 hours of injection. RESULTS: Labeling efficiency of immature and mature DC were 60.4 +/- 5.4% and 61.8 +/- 6.7%, respectively. Iv-DC initially appeared in the lungs, then redistributed mainly to liver and spleen. Migration of mature DC to spleen was significantly higher than that of immature DC (38.3 +/- 4.0 % vs. 32.2 +/- 4.1 % in control group, 40.4 +/- 4.1% vs. 35.9 +/- 3.8 % in tumor group; p< 0.05). Migration to tumor was also significantly higher in mature DC than in immature DC (2.4 +/- 0.3% vs 1.7 +/- 0.2%; p=0.034). CONCLUSION: Assessment of migration pattern of DC in mice was possible using 99mTc-HMPAO labeled immature and mature DC. Migration of mature DC to spleen and tumor was higher than that of immature DC when they were i.v. injected.

Development and Assessment Individual Maximum Permissible Dose Method of I-131 Therapy in High Risk Patients with Differentiated Papillary Thyroid Cancer / 대한핵의학회잡지

PURPOSE: Radioiodine (I-131) therapy is an effective modality to reduce both recurrence and mortality rates in differentiated thyroid cancer. Whether higher doses shows higher therapeutic responses was still debatable. The purpose of this study was to validate curve-fitting (CF) method measuring maximum permissible dose (MPD) by a biological dosimetry using metaphase analysis of peripheral blood lymphocytes. MATERIALS AND METHODS: Therapeutic effects of MPD was evaluated in 58 patients (49 females and 9 males, mean age 50+/-11 years) of papillary thyroid cancer. Among them 43 patients were treated with or =9.25 GBq. The former was defined as low-dose group, and the latter high-dose group. Therapeutic response was defined as complete response when complete disappearance of lesions on follow-up I-131 scan and undetectable serum thyroglobulin levels were found. Statistical comparison between groups were done using chi-square test. P value less than 0.05 was regarded as statistically significant. RESULTS: MPD measured by CF method using tracer and therapeutic doses were 13.3+/-1.9 and 13.8+/-2.1 GBq, respectively (p=0.20). They showed a significant correlation (r=0.8, p< 0.0001). Exposed doses to blood measured by CF and biological methods were 1.54+/-0.03 and 1.78+/-0.03 Gy (p=0.01). They also showed a significant correlation (r=0.86, p=0.01). High-dose group showed a significantly higher rate of complete response (12/15, 80%) as compared to the low-dose group (22/43, 51.2%) (p=0.05). While occurrence of side effects was not different between two groups (40% vs. 30.2%, p=0.46). CONCLUSION: Measurement of MPD using CF method is reliable, and the high-dose I-131 therapy using MPD gains significantly higher therapeutic effects as compared with low-dose therapy.

Radioresistance of Dendritic Cells / 대한핵의학회잡지

PURPOSE: To evaluate radiation sensitivity of dendritic cells in comparison with lymphocytes. MATERIALS AND METHODS: T lymphocytes captured from peripheral blood were irradiated by 0 Gy, 10 Gy, 30 Gy. Apoptosis was measured by flowcytometry for staining of Annexin V 4 hours after irradiation. Immature and mature dendritic cells processed from blood hematopoietic stem cell were irradiated by 0 Gy, 10 Gy, 30 Gy, 100 Gy respectively and apoptosis was measured by flowcytometry with time difference as 4h, 24h and 48h after irradiation. Morphometric analysis by percent nucleus was measured in three cell groups, also. RESULTS: Lymphocytes showed radiation sensitivity by increasing apoptotic fraction according to radiation dose. However, both mature and immature dendritic cells showed consistent fraction of apoptosis in spite of increasing radiation dose. Percent nucleus ratio is significantly higher in lymphocytes than that of mature or immature dendritic cells. Stimulation of T-cell by dendritic cells was not changed after irradiation. CONCLUSION: Dendritic cells showed radioresistance which was associated with small size of nucleus in comparison with lymphocytes and this result would be used as a basal data of radio-labelling for the cellular trafficking studies in nuclear medicine fields.

Radiation Adaptive Response Induced by I-131 Therapy in Patients with Differentiated Thyroid Cancer / 대한핵의학회잡지

PURPOSE: The purpose of this study was to ascertain whether radiation adaptive response could be induced by high dose I-131 therapy in patients with differentiated thyroid cancer. MATERIALS AND METHODS: Lymphocytes from 21 patients (7 males, 14 females, mean age 55+/-12 years) were collected before and after administration of 5,550 MBq (150 mCi) I-131. They were exposed to a challenge dose of 1 Gy gamma rays using a Cs-137 cell irradiator. The number of ring-form (R) and dicentric (D) chromosomes was counted under the light microscope, and used to calculate the frequency of chromosomal aberration. Ydr, which was defined as the sum of R and D divided by the total number of counted lymphocytes. RESULTS: Ydr in patients before I-131 therapy (0.09+/-0.01) was not different from that of controls (0.08+/-0.01). Ydr was significantly increased to 0.13+/-0.02 (p<0.0001) after I-131 therapy. Increase of Ydr after the challenge irradiation of 1 Gy was significantly lower in patients after I-131 therapy than before I-131 therapy (0.17+/-0.03 vs 0.21+/-0.02, p<0.0001). Cycloheximide (CHM), an inhibitor of protein synthesis, abolished this effect. Ydr after CHM (0.20+/-0.01) was significantly higher than Ydr after I-131 therapy (0.17+/-0.03, p<0.0001), but was not different from Ydr before I-131 therapy (0.21+/-0.02). CONCLUSION: High dose I-131 therapy induces an adaptive response in peripheral lymphocytes of patients with well-differentiated thyroid cancer, which is associated with protein synthesis.

Comparison of Radiation Adaptive Responses in Peripheral Lymphocytes of Patients Undergoing Tc-99m MDP and Tc-99m DTPA Scintigraphies / 대한핵의학회잡지

PURPOSE: The purpose of this study was to compare the radiation adaptive response (RAR) in peripheral lymphocytes (PL) of patients induced by Tc.-99m MDP and Tc-99m DTPA scintigraphies. MATERIALS AND METHODS: Lymphocytes from 45 patients (25 males, 20 females, mean age 44+/-18 years) were collected before and after scintigraphies using 740 MBq Tc-99m MDP (n=22) or Tc-99m L)TPA (n=23). Lympho-cytes from 20 controls (12 males, 8 females, mean age 43+/-7 years) were also callected. They were exposed to challenge dose of 2 Gy gamma-rays using a Cs-137 cell irradiator, Number of ring-form (R) and dicentric (D) chromosomes was counted under the light microscope. From them a representative score, Ydr, was calculated as Ydr=(D+R)/cells. Adaptation index (AI) was defined as difference of Ydr between unconditioned and conditioned lymphocytes. Ydr was also measured after an administration of cyclohexi-mide (CHM), a protein synthesis inhibitor, before challenge dose. RESULTS: RAR was induced in both groups of patients. CHM abolished the adaptive response in both groups. AI of Tc-99m MDP group was significantly higher than that of Tc-99m DTPA group. CONCLUSION: Tc-99m MDP induced RAR was more prominent than those induced by Tc-99m DTPA.