Recent progress on betulinic acid and its derivatives as antitumor agents: a mini review / 中国天然药物

Natural products are one of the important sources for the discovery of new drugs. Betulinic acid (BA), a pentacyclic triterpenoid widely distributed in the plant kingdom, exhibits powerful biological effects, including antitumor activity against various types of cancer cells. A considerable number of BA derivatives have been designed and prepared to remove their disadvantages, such as poor water solubility and low bioavailability. This review summarizes the current studies of the structural diversity of antitumor BA derivatives within the last five years, which provides prospects for further research on the structural modification of betulinic acid.

Research progress on pyruvate dehydrogenase kinase inhibitors / 药学学报

Tumor cells can metabolize glucose through glycolysis to intermediates for biomacromolecule synthesis by inhibiting the activity of the pyruvate dehydrogenase complex (PDC) in mitochondria. In this process, pyruvate dehydrogenase kinases (PDKs) play a key role. The inhibition of the activity of PDKs can effectively block this metabolic pathway, activate mitochondrial oxidative metabolism, and induce tumor cell apoptosis. PDK inhibitors have become a research hotspot in medicinal chemistry, and novel structures targeting classical binding sites have been synthesized. In this paper, recent research progress on PDK inhibitors is reviewed to provide information on these latest entities and to explore their clinical applicability.

Silencing the FOXQ1 gene inhibits the proliferation of thyroid papillary carcinoma TPC-1 cells / 医学研究生学报

Objective Forkhead box Q1 (FOXQ1) is highly expressed but its biological role remains unclear in papillary thyroid carcinoma (PTC). This article aims to investigate the effect of FOXQ1-siRAN on the proliferation of TPC-1 cells and its possible mechanism.Methods Synthetic FOXQ1-siRNA and NC-siRNA were transfected into TPC-1 cells by lipofectamineTM2000. The cells were divided into five groups: NC-siRAN, FOXQ1-1, FOXQ1-2, FOXQ1-3 and blank control. After transfection, the expressions of the FOXQ1 mRNA and protein, as well as those of c-Myc and cyclinD1 proteins, were determined by qRT-PCR and Western blot, and the changes in the proliferation of the TPC-1 cells observed by MTT.Results Compared with the blank control and NC-siRAN groups, the FOXQ1-3 group showed a significantly inhibited expression of FOXQ1 mRNA (P<0.05). The protein expression of FOXQ1 was markedly decreased in the FOXQ1-1, FOXQ1-2 and FOXQ1-3 groups in comparison with that in the NC-siRAN group (0.54±0.07, 0.40±0.07 and 0.26±0.06 vs 0.78±0.08, P<0.05). The proliferation of the TPC-1 cells was remarkably lower in the FOXQ1-3 than in the blank control and NC-siRNA groups (P<0.05), and so were the relative protein expressions of c-Myc (0.57±0.04 vs 1.05±0.07 and 0.92±0.06, P<0.05) and cyclinD1 (0.51±0.10 vs 0.94±0.12 and 0.91±0.11,P<0.05).Conclusion Silencing the FOXQ1 gene can effectively inhibit the proliferation of TPC-1 cells in TPC, probably by suppressing the expressions of c-Myc and cyclinD1.

Inhibition of ATM Promotes Neuronal Apoptosis through Upregulating Bim Expression / 中山大学学报(医学科学版)

@#【Abstract】 【Objective】To investigate the mechanisms implicated in ataxia telangiectasia mutated(ATM)inhibitioncaused apoptosis in the cultured cerebellar granule neurons.【Methods】Primary cerebellar granule neurons(CGN)isolated from neonatal Sprague Dawley rats of 7-8 days were divided into the following groups:25 K group(survival group),5 K group(apoptosis group)and 25 K + KU-55933 treatment group(ATM inhibition group),25 K + KU-55933 + Mithramycin A treatment group(MMA group),25 K + KU-55933 + Chromonycin A3 treatment group(CMA3 group). The protein expression of p-ATM,ATM,Bim and Caspase 3 were detected by Western Blot. The apoptotic cells with nuclear pyknosis were detected by Hoechst-staining.【Results】Compared with 25 K group,the result of western blot showed that the protein expression of Bim and Caspase 3 were increased in the ATM inhibition group(P < 0.05). Compared with 25 K group,the nuclear pyknosis rate of 5 K group and ATM inhibition group were significantly increased(P < 0.05). Inhibition of Bim by Mithramycin A or Chromomycin A3 remarkably reversed ATM inhibition-caused apoptosis.【Conclusion】Inhibition of ATM induce Bim dependent apoptosis in cultured cerebellar granule neurons.

Effects of low-intensity pulsed ultrasound on fracture healing and caveolin-1 gene expression in the radius defects of rabbits / 复旦学报（医学版）

Objective To explore the effect of low-intensity pulsed ultrasound (LIPUS) on long bone fracture healing and to examine caveolin-1 gene expression in the radius defects of rabbits.Methods A total of 24 New Zealand rabbits with 3-mm bone defects at lower 1/3 in both radii were randomly assigned to 4 groups (n=6).Daily LIPUS treatment was performed to the right fracture sites at a intensity of 30 mW/cm2 for 20 minutes,while the left sites received sham treatment with power off.To assess the effects of LIPUS on bone defects,X-ray imaging and hematoxylin-eosin staining were applied 7,14,21,28 days after the surgery.Additionally,the immunohistochemical staining was used to determine the subcllular localization of caveolin-1 and semi-quantify the caveolin-1 level,qPCR was performed to detect the mRNA level of caveolin-1,gene Col2a1 and Col10a1,and osteocalcin.Results On day 14,the radiological score of the right radii and mineralized callus area were significantly higher than that of the left ones,both of them were elevated with time flied.Histological examination suggested that the differentiation and apoptosis of chondrocytes along with the formation and bridging of the bone trabeculas appeared earlier in the right radius defects.The immunohistochemical staining showed that on day 7 and 14,the level of caveolin-1 increased with the proliferation and differentiation of condrocytes,and was significantly higher in callus tissues on the right sites.On day 21 and 28,the mesenchymal stem cells migrated to the surface of cartilage matrix started to differentiate into osteoblasts,the level of caveolin-1 decreased,and was significantly lower on the right sites.The result of qPCR indicated that compared with the left sites the caveolin-1 gene expression on the right sites was significantly higher on day 7,while significantly lower on day 21.The mRNA expression levels of Col2a1,Col10a1 and osteocalcin on the right sites were significantly higher on day 7 and 14,but they were significantly lower on day 21 and 28,except for Col10a1 on day 28.Conclusions Advancing endochondral ossification is considered to be a crucial mechanism during long bone fracture healing promoted by LIPUS.The caveolin-1 gene expression first increased in the chondrocytes then decreased in the mesenchymal stem cells during the process.

In vivo biomechanical study of cervical spine kinematics / 医用生物力学

The traditional method of in vitro biomechanical study cannot simulate the realistic environment in human body due to the limitation of in vitro technology. Using advanced imaging techniques and testing methods, in vivo biomechanical studies of cervical spine kinematics can directly observe the cervical spine motion of living subjects. The data obtain repeats physiological situations and has important implications for improvement in clinical practice. In this paper, the biomechanical research methods of cervical spine kinematics, as well as the development process and current status were reviewed. The postoperative changes in cervical spine kinematics and the effects on adjacent segments following cervical spine fusion and cervical disc arthroplasty surgery were analyzed. The future developments in cervical spine research were also discussed.

Role of glucose-regulated protein 78 in early brain injury after experimental subarachnoid hemorrhage in rats / 华中科技大学学报（医学）（英德文版）

Early brain injury (EBI) plays a key role in the pathogenesis of subarachnoid hemorrhage (SAH). This study investigated the role of glucose-regulated protein 78 (GRP78) in EBI after SAH. Male Sprague-Dawley rats (n=108) weighing 260±40 g were divided into control, sham-operated, and operated groups. Blood was injected into the prechiasmatic cistern of rats in the operated group. Neurological scores, ultrastructures of neurons, apoptosis, and GRP78 expression in the hippocampus were examined using Garcia scoring system, transmission electron microscopy, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling, and Western blotting at 1, 6, 12, 24, 48, and 72 h after SAH, respectively. The results showed that neurological scores were significantly decreased in the operated group as compared with those in control and sham-operated groups at 12, 24, 48, and 72 h. Metachromatin, chromatin pyknosis at the edge, endoplasmic reticulum swelling, and invagination of nuclear membrane were observed at 24 h in the operated group, indicating the early morphological changes of apoptosis. The number of apoptotic cells was significantly increased in the operated group as compared with that in control and sham-operated groups at 6, 12, 24, 48, and 72 h. The GRP78 protein expression levels in the operated group were significantly elevated at all time points and reached the peak at 12 h. GRP78 expression was positively associated with apoptosis cells and negatively with neurological scores. In conclusion, EBI was demonstrated to occur after SAH and GRP78 was involved in the development of EBI after SAH.

Effectiveness of CLAT Protocol for Treating Patients with Refractory Acute Myeloid Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical efficacy and toxicity of CLAT protocol (cladribine, cytarabine and topotecan) for treating patients with refractory acute myeloid leukemia (R-AML).</p><p><b>METHODS</b>A total of 18 patients with R-AML (median age 37 years, range 18 to 58 years; male n = 16, female n = 2) were treated with CLAT protocol, which consisted of cladribine 5 mg/m(2)/d, i.v. on days 1-5, cytarabine 1.5 g/m(2)/d, i.v. on days 1-5, topotecan 1.25 mg/m(2)/d, i.v. on days 1-5 and G-CSF 300 µg/d subcutaneous injection on day 6 until neutrophile granulocyte recovery.</p><p><b>RESULTS</b>Out of 18 patients 2 died of severe infection before the assessment. Among 16 evaluated patients, 10 (55.6%) achieved complete remission (CR), and 2 (11.1%) achieved partial remission (PR), the overall response rate was 66.7%, the rest 4 patients did not respond (NR). The median overall survival time and DFS for the CR patients was 9.5 months (95%CI: 6.7-16.64) and 9.5 months (95%CI: 6.1-16.7) respectively. The 1 year OS and DFS rates were 45% and 46.9%, respectively. All patients developed grade 4 of granulocytopenia and thrombocytopenia, the median duration was 13 (range 2 to 21) days and 12 days (range 2 to 21), respectively, all patients developed infection, 2 patients died of severe infection. The most common non-hematological side effects included nausea, vomiting, diarrhoea, rash, aminotransferase or bilirubin elevation and were grade 1 to 2.</p><p><b>CONCLUSION</b>The CLAT protocol seems to have promising for the treatment of refractory AML patients, and patients well tolerated. This CLAT protocol offers an alternative treatment for R-AML patients who received severe intensive treatment, especially with anthracycline-containing chemotherapy.</p>

Role of glucose-regulated protein 78 in early brain injury after experimental subarachnoid hemorrhage in rats / 华中科技大学学报（医学）（英德文版）

Early brain injury (EBI) plays a key role in the pathogenesis of subarachnoid hemorrhage (SAH). This study investigated the role of glucose-regulated protein 78 (GRP78) in EBI after SAH. Male Sprague-Dawley rats (n=108) weighing 260±40 g were divided into control, sham-operated, and operated groups. Blood was injected into the prechiasmatic cistern of rats in the operated group. Neurological scores, ultrastructures of neurons, apoptosis, and GRP78 expression in the hippocampus were examined using Garcia scoring system, transmission electron microscopy, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling, and Western blotting at 1, 6, 12, 24, 48, and 72 h after SAH, respectively. The results showed that neurological scores were significantly decreased in the operated group as compared with those in control and sham-operated groups at 12, 24, 48, and 72 h. Metachromatin, chromatin pyknosis at the edge, endoplasmic reticulum swelling, and invagination of nuclear membrane were observed at 24 h in the operated group, indicating the early morphological changes of apoptosis. The number of apoptotic cells was significantly increased in the operated group as compared with that in control and sham-operated groups at 6, 12, 24, 48, and 72 h. The GRP78 protein expression levels in the operated group were significantly elevated at all time points and reached the peak at 12 h. GRP78 expression was positively associated with apoptosis cells and negatively with neurological scores. In conclusion, EBI was demonstrated to occur after SAH and GRP78 was involved in the development of EBI after SAH.

Preliminary analysis of hippocampal volume measurement based on magnetic resonance imaging in 68 cases / 实用医学杂志

Objective To investigate the volume of hippocampal formation in normal adult using coronal magnetic resonance imaging. Methods 3D-fSPGR sequence was used to depict the brain in 68 healthy adult. The volume of hippocampus was calculated by drawing the outline of 10 coronal hippocampal formation images acquired equally form posterior border of rostrum corpus callosum to anterior border of the splenium. Data were analysed using the SPSS 17.0 software. Results In the coronal plane images of normal adult brain, the absolute volume of left and right hippocampal formation were 2 319.63-2 610.73 mm3 and 2 447.52-2 749.50 mm3 respectively . The relative volume of left and right hippocampal formation were 2 319 . 87-2 602 . 47 mm 3 and 2 443.96-2 755.89 mm3. There were no correlation between hippocampal volume and age (r = 0.084, P = 0.549. Significant gender differences (t=2.500, P=0.029) were observed between absolute volume of right hippocampal formation in the youth group. There were significant differences in the absolute volume (t = -2.571, P = 0.022), relative volume (t = 2.600, P = 0.021) among the right and left hippocampal formation. Significant absolute volume differences (P = 0.038) were observed between the middle-aged group and the youth group among the hippocampal formation of women. Conclusion No significant differences were observed in age, gender among the hippocampal volume of normal adult, and there was a significant difference between the left and right hippocampal formation volume.

The clinical manifestations of olivopontocerebellar atrophy and the diagnostic value of MR / 中国医师进修杂志

Objective To investigate the clinical manifestations and MR imaging features of olivopontocerebellar atrophy(OPCA).Methods The clinical data and MR findings of 8 patients with OPCA (OPCA group) were retrospectively analyzed,and 75 cases of healthy physical examination people were chosen as control group (they were divided into 21-30 years group,31-40 years group,41-50 years group,51-60 years group,61-70 years group,and 15 cases in each group),all subjects under went pons measurement.Results All 8 patients with OPCA were sporadic cases.The cerebellar symptom,autonomic symptoms and extrapyramidal symptoms were common manifestations.The head MR imaging of OPCA presented with cerebellum and brain stem atrophy,cross sign was the characteristic appearance.Vertical diameter and anteroposterior diameter of the pons in OPCA group patients was (20.63 ± 2.16) and (16.74 ±4.37) mm,compared with control group [(27.14 ±1.73) and (22.28 ± 1.49) mm],there was significant difference (P ＜ 0.01).In control group,vertical diameter and anteroposterior diameter of the pons maximum in 21-30 years group and 31-40 years group,increased with age,the value had been reduced,but there was no significant difference (P ＞ 0.05).Conclusion MR is the first choice for the diagnosis of OPCA,combine with clinical manifestations may improve the diagnostic accuracy.

C-kit mutation in acute myeloid leukemia patients with AML1-ETO fusion gene and its clinical significance / 中国实验血液学杂志

This study was aimed to investigate the c-kit mutation in acute myeloid leukemia (AML) patients with AML1-ETO and analyze its relation with clinical and laboratorial features and prognosis. PCR and sequencing methods were used to detect the c-kit 17 exon mutations in 31 AML patients with AML1-ETO. The relation of the c-kit mutation with clinical features, results of laboratorial examination and prognosis of disease were analyzed. The results showed that the c-kit mutation was found in 14 out of 31 AML patients and the mutation frequency was 45.16%. Male patients had a higher incidence of c-kit mutation than that of female patients (P = 0.020). The proportion of patients with newly diagnosed white blood cell>10×10(9)/L and with extramedullary infiltration in mutated group were higher than those in unmutated group respectively. No significant difference was observed at the age (P = 0.437) and the rate of bone marrow blasts(P = 0.510) between the above mentioned two groups. The difference in complete remission rate (64.29% vs 80%, P = 0.344)and relapse rate (58.33% vs 21.43%, P = 0.054) between c-kit mutated and c-kit unmutated groups were not significant. While the c-kit mutated group had a significant higher death rate as compared with c-kit unmutated group (57.14% vs 20%, P = 0.039). It is concluded that the c-kit mutation is frequent in AML patients with AML1-ETO and the c-kit mutated patients have a poor prognosis. It is important to detect c-kit mutation in routine clinical practice for patient's risk stratification, evaluation of prognosis and selection of effective treatment.

Surgical treatment of Siewert II adenocarcinoma of the esophagogastric junction / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the outcomes after surgical treatment of esophagogastric junction carcinoma (EGJC).</p><p><b>METHODS</b>One hundred and eighty-five patients with EGJC undergoing surgery from October 2000 to September 2006 at the Cancer Hospital of Shantou University were reviewed retrospectively. The clinical outcomes were compared between transthoracic and transabdominal approach.</p><p><b>RESULTS</b>Of the 185 patients, 133 underwent operation via transthoracic approach and 52 via transabdominal approach. The postoperative complication rates were 10.5%(14/133) and 11.5%(6/52) and the 1-, 3-, 5-year overall survival rates were 83.9%, 44.5%, 32.9% and 86.0%, 38.0%, 30.0% in transthoracic and transabdominal groups respectively, and the difference were not statistically significant (both P>0.05).</p><p><b>CONCLUSION</b>Surgical approach should be individualized for EGCJ.</p>

Detection of methylation levels of multi-genes by real-time PCR in patients with myelodysplastic syndrome / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze the promoter methylation levels of p15, CDH1, DAPK and HICI genes of patients with myelodysplastic syndrome (MDS) and explore the relationship between the level of methylation and clinical features.</p><p><b>METHODS</b>DNA methylation levels of p15, CDH1, DAPK and HICI in peripheral blood (PB) or bone marrow (BM) samples from 52 MDS patients were detected by real-time quantitative PCR. The correlation of the methylation level with clinical features and hematological findings was analyzed. 38 de novo AML patients and 46 normal individuals served as controls.</p><p><b>RESULTS</b>The methylation levels of p15, CDH1, DAPK and HICI were 16.23 ± 21.69, 6.59 ± 9.39, 0.14 ± 0.11 and 7.81 ± 9.70 in BM, and 14.96 ± 20.16, 6.00 ± 9.26, 0.12 ± 0.14 and 6.74 ± 9.72 in PB, respectively from 18 MDS patients, and the difference between BM and PB was not statistically significant (P > 0.05). The methylation levels of p15 (14.70 ± 18.17) and CDH1 (6.61 ± 8.79) genes in high risk (RAEBI/II) MDS were significantly higher than in low risk (RCMD/RARS/5q-, p15: 1.99 ± 1.59, CDH1: 1.23 ± 1.14 and RCMD, p15: 3.02 ± 3.42, CDH1:1.53 ± 2.06) MDS or control (p15: 1.69 ± 1.82, CDH1: 1.01 ± 1.12) (P < 0.05). The methylation levels of DAPK gene had no difference among subtypes of MDS, and that of HIC1 gene only differed between RAEB I/II (9.16 ± 11.95) and control (2.49 ± 2.26) (P = 0.042). The difference of methylation levels of p15, CDH1, DAPK and HICI in BM was statistically significant among subtypes of MDS (P = 0.001, 0.003, 0.039, 0.023, respectively). And so did of p15 and DAPK in PB (P = 0.013, 0.006, respectively). The methylation level of p15 and CDH1 was significantly correlated with IPSS classification and blasts percentage in BM.</p><p><b>CONCLUSIONS</b>p15 and CDH1 genes are special hypermethylation genes in MDS. Methylation level of HIC1 gene showed an upward tendency from low risk to high risk MDS.</p>

Expression of CD133 in the bone marrow of patients with myelodysplastic syndrome and its clinical significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of CD133 in the bone marrow of patients with myelodysplastic syndrome (MDS) and explore its clinical significance.</p><p><b>METHODS</b>The expression of CD133 and CD34/CD38 in the bone marrow was detected using flow cytometry in 31 cases of refractory anemia with excess blasts (RAEB), 10 cases of refractory cytopenia with multilineage dysplasia (RCMD) and 11 cases of aplastic anemia (AA).</p><p><b>RESULTS</b>The percentage of CD133-expressing cells was 6.75% in patients with RAEB, significantly higher than that in patients with RCMD (1.41%) and AA (2.70%) (P<0.05); the percentage of CD133-positive cells were similar between the latter two patient groups (P>0.05). The percentage of CD34(+)/CD38- cells was similar in the 3 groups (P>0.05), all lower than 1%.</p><p><b>CONCLUSIONS</b>Advanced MDS patients are characterized by an increase of CD133-expressing cells, suggesting the value of CD133 in the diagnosis of RAEB. CD34(+)/CD38- cells do not show a significant value in the diagnosis of MDS.</p>

Serum proteomics in patients with RAEB myelodysplastic syndromes / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen the molecular markers for refractory anemia with excess blasts in transformation (RAEB) in myelodysplastic syndromes (MDS) by serum proteome profiling.</p><p><b>METHODS</b>The serum protein were isolated from patients with RAEB, acute myeloid leukemia or normal subjects by 2-dimensional electrophoresis (2-DE), and the electrophoresis gels were obtained to identify the differentially reacting protein spots. The replica gels of the differentially reacting proteins were analyzed to locate the matching protein spots, which were identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOF-MS) and database searching.</p><p><b>RESULTS</b>Seven differentially expressed proteins in RAEB were found by 2-DE. Of the 7 proteins, 4 were identified by MALDI-TOF-MS to have significantly differential expression in RAEB, including dipeptidyl peptidase (DPP/CD26), polymerase (DNA directed) kappa, PRO2044 and an albumin-like protein.</p><p><b>CONCLUSION</b>2-DE-based serum proteome profiling helps identify serum proteomic biomarkers related to MDS. DDP/CD26 has increased expression in the serum in RAEB subtype MDS, suggesting its possible role in advanced MDS.</p>

Expression of antiapoptotic gene aven in de novo acute myeloid leukemia patients and its clinical significance / 中国实验血液学杂志

This study was aimed to investigate the aven mRNA expression level of leukocytes from peripheral blood(PB)of de novo patients with acute myeloid leukemia (AML) and analyze its clinical significance, so as to provide a experimental basis for evaluating prognosis. The aven mRNA expression levels in PB samples from 69 de novo AML patients were detected by using real-time quantitative PCR. The relation of aven mRNA level with clinical and hematological characteristics (age, sex, WBC, Hb, Plt, LDH, Blast% in PB and BM, FAB subtype) and treatment outcome (CR rate and relapse rate) were analyzed. 21 normal individuals served as controls. The results showed that the expression level of aven mRNA was between 11.72% and 178.93% (median 37.2%) in de novo AML and between 10.81% and 50.98% (median 28.81%) in normal individuals. Aven mRNA expression level was higher in the AML group than that in the controls (p = 0.006). As aven mRNA expression level was compared with other clinical and hematological parameters, there were significant correlations between aven mRNA expression level and age (r = 0.25, p = 0.039), and between hemoglobin level (r = 0.29, p = 0.019), FAB subtype(r = 0.253, p = 0.036). The median expression level (50.08%) of aven mRNA in older patients (> or = 44 years) was higher then that (32.41%) in younger patients (< 44 years) (p = 0.018). The complete remission (CR) rate after two cycles of chemotherapy in patients with lower aven mRNA level (25/30, 83.33%) was higher than that in patients with higher aven mRNA level(21/30, 70%), but the difference was not significant(p = 0.22). The difference of aven mRNA expression level between AML patients with relapse and AML patents without relapse was not significant (p = 0.076). It is concluded that the expression level of aven mRNA in de novo AML patients obviously increases, the overexpression of aven mRNA likely involves in genesis of AML. The definite relation of aven mRNA expression level with treatment outcome and relapse was not been found.