Synergistic antitumor activity of artesunate and HDAC inhibitors through elevating heme synthesis synergistic upregulation of ALAS1 expression

Artemisinin and its derivatives (ARTs) were reported to display heme-dependent antitumor activity. On the other hand, histone deacetylase inhibitors (HDACi) were known to be able to promote heme synthesis in erythroid cells. Nevertheless, the effect of HDACi on heme homeostasis in non-erythrocytes remains unknown. We envisioned that the combination of HDACi and artesunate (ARS) might have synergistic antitumor activity through modulating heme synthesis. studies revealed that combination of ARS and HDACi exerted synergistic tumor inhibition by inducing cell death. Moreover, this combination exhibited more effective antitumor activity than either ARS or HDACi monotherapy in xenograft models without apparent toxicity. Importantly, mechanistic studies revealed that HDACi coordinated with ARS to increase 5-aminolevulinate synthase (ALAS1) expression, and subsequent heme production, leading to enhanced cytotoxicity of ARS. Notably, knocking down significantly blunted the synergistic effect of ARS and HDACi on tumor inhibition, indicating a critical role of ALAS1 upregulation in mediating ARS cytotoxicity. Collectively, our study revealed the mechanism of synergistic antitumor action of ARS and HDACi. This finding indicates that modulation of heme synthesis pathway by the combination based on ARTs and other heme synthesis modulators represents a promising therapeutic approach to solid tumors.

Expression of CYP4A11 and CYP4A22 in triple negative breast carcinoma and its relation to M2 tumor-associated macrophages / 临床与实验病理学杂志

Purpose To investigate the expression of CYP4 A11 and CYP4 A22 in triple-negative breast carcinoma (TNBC) and its relationship with clinicopathological features and M2 tumor-associated macrophages (TAMs). Methods 72 cases of TNBC with clinical and pathological data were collected. The expression of CYP4 A11 and CYP4 A22 in the carcinoma cells and the expression of CD68 and CD163 of the TAMs were detected by immunohistochemically and analyzed with image processing software. The relationship between the expressions of CYP4 A11 and CYP4 A22 with clinicopathologic features and its correlation of the M2 state of TAMs was studied. Results Both the immunohistochemically staining scores of CYP4 A11 and CYP4 A22 were higher in cancer tissues than that in breast tissues (P<0.001, P<0.001). The higher expression of CYP4 A11 was associated with tumor diameter increase (P<0.001), lymph node metastasis (P<0.001), higher clinical stage (P<0.001) and higher Ki-67 index (P=0.011). Both the positive rates of CD68 and CD163 in the high expression group of CYP4 A11 were higher than those in the low expression group of CYP4 A11 (P=0.021, P<0.001). The higher expression of CYP4 A22 was associated with lymph node metastasis (P<0.001), higher clinical stage (P=0.006), higher recurrence rate (P<0.001), and higher Ki-67 index (P=0.040).The positive rates of CD163 in the high expression group of CYP4 A22 was higher than that in the low expression group of CYP4 A22 (P<0.001). Conclusion Both the expression of CYP4 A11 and CYP4 A22 may be associated with M2 polarization state of TAMs, high proliferative activity and lymph node metastasis in the TNBC.

Lipid-lowering effects of oleanolic acid in hyperlipidemic patients / 中国天然药物

Oleanolic acid (OA) is a pentacyclic triterpenoid compound extracted from olea europaeal, a traditional Chinese medicine herb. OA has been used in the clinic as a hepatoprotective medicine in China since 1970s. In our previous study, we observed that OA could ameliorate hyperlipidemia in animal models. In the present study, we conducted a small-scale clinical trial to evaluate the hypolipidemia effect of OA in hyperlipidemic patients. Hyperlipidemic patients were administrated with OA for four weeks (4 tablets once, three times a day). The blood samples of the patients were collected before and after OA treatment. The biological parameters were measured. Furthermore, three patients' blood samples were studied with DNA microarray. After OA administration, the TC, TG, and HDLC levels in serum decreased significantly. DNA microarray analysis results showed that the expressions of 21 mRNAs were significantly changed after OA treatment. Bioinformatics analysis showed 17 mRNAs were up-regulated and 4 mRNAs were down-regulated significantly after OA treatment. Five mRNAs (CACNA1B, FCN, STEAP3, AMPH, and NR6A1) were selected to validate the expression levels by qRT-PCR. Therefore, OA administration differentially regulated the expression of genes involved in lipid metabolism. The data showed a clinical evidence that OA could improve hyperlipidemia and also unveiled a new insight into the molecular mechanisms underlying the pharmacological effect of OA on hyperlipidemia.

Lipid-lowering effects of oleanolic acid in hyperlipidemic patients / 中国天然药物

Oleanolic acid (OA) is a pentacyclic triterpenoid compound extracted from olea europaeal, a traditional Chinese medicine herb. OA has been used in the clinic as a hepatoprotective medicine in China since 1970s. In our previous study, we observed that OA could ameliorate hyperlipidemia in animal models. In the present study, we conducted a small-scale clinical trial to evaluate the hypolipidemia effect of OA in hyperlipidemic patients. Hyperlipidemic patients were administrated with OA for four weeks (4 tablets once, three times a day). The blood samples of the patients were collected before and after OA treatment. The biological parameters were measured. Furthermore, three patients' blood samples were studied with DNA microarray. After OA administration, the TC, TG, and HDLC levels in serum decreased significantly. DNA microarray analysis results showed that the expressions of 21 mRNAs were significantly changed after OA treatment. Bioinformatics analysis showed 17 mRNAs were up-regulated and 4 mRNAs were down-regulated significantly after OA treatment. Five mRNAs (CACNA1B, FCN, STEAP3, AMPH, and NR6A1) were selected to validate the expression levels by qRT-PCR. Therefore, OA administration differentially regulated the expression of genes involved in lipid metabolism. The data showed a clinical evidence that OA could improve hyperlipidemia and also unveiled a new insight into the molecular mechanisms underlying the pharmacological effect of OA on hyperlipidemia.

Expression of S1PRl,S1PR4 in triple negative breast carcinoma and its significance / 临床与实验病理学杂志

Purpose To investigate the expression of sphingosine-1 phosphate receptor 1 (S1PR1) and sphingosine-1 phosphate receptor 4 (SIPR4) in triple negative breast carcinoma (TNBC) and to evaluate its correlation with the clinicopathologic features of TNBC. Methods 72 cases of tissue slides of TNBC were stained immunohistochemically and analyzed with image processing to calculate the S1PR1 and S1PR4 expression. Correlations of the S1PR1 and S1PR4 expression with the clinicopathologic features of TNBC were studied. Results Ki-67 index of high, moderate and low expression of the S1PR1 in TNBC were 48.89％, 36.11％ and 26.48％, respectively. The difference among them was significance (P＜0.001). Ki-67 index of high, moderate and low expression of the S1PR4 in TNBC were42.83％, 31.43％ and 28.93％, respectively. The difference among them was significance (P = 0.007 ). The positive rate of lymph node of high, moderate and low expression of the SI PR1 in TNBC were 31.4％, 48.6％ and 20.0％, respectively. The difference among them was significance (P = 0.012). The positive rate of lymph node of high, moderate and low expression of the S1PR4 in TNBC were 54.3％, 40.0％ and 5.7％, respectively. The difference among them was significance (P=0.010). The CD68 positive rate of high, moderate and low expression of the S1PR1 in TNBC were 47.22％, 42.59％ and31.48％, respectively. The difference among them was significance (P = 0.036). Both the difference of survive rate among high, moderate and low expression of the S1PR1 and S1PR4 were not significance (P = 0.209 and P =0.593 ). Conclusion High expression of S1PR1 and S1PR4 may contribute to the cellular proliferation and lymph node metastases in TNBC. The survive rate of TNBC maybe not related with both the S1PR1 and S1PR4 expression.

Clinical and pathological features in children with progressive muscular dystrophy / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the clinical and pathological features of progressive muscular dystrophy (PMD) in children and to provide help for the early and accurate diagnosis of PMD.</p><p><b>METHODS</b>Retrospective analysis was performed on the clinical data of 99 hospitalized children with PMD, including clinical manifestations, age of onset, family history, creatase, electromyogram (EMG) and pathological changes of muscles.</p><p><b>RESULTS</b>Of the 99 children with PMD, the age of onset was 0.5-14.5 (4.7 ± 3.1) years. Eleven cases (11%) had a family history of PMD. Twenty-six (26%) were misdiagnosed as other diseases. All patients presented with muscle weakness when seeing the doctor, and 66 (67%) of them had muscle atrophy and/or hypertrophy. All patients had elevated creatine kinase (CK) levels. The 2-7-year-old group (n=51) had a mean CK level of 9965 ± 8876 U/L, and the 7-15-year-old group (n=48) had a mean CK level of 5110 ± 4498 U/L, with a significant difference between the two groups (P<0.01). The EMG examination performed on 66 patients showed that 54 cases (82%) had myogenic damage and 10 cases (15%) had neurogenic damage. Light microscopy revealed coexistence of atrophy and hypertrophy of muscle fibers, hyaline degeneration and granular degeneration. Electron microscopy showed that muscle fibers were different in thickness, some atrophic or hypertrophic; muscle cell nuclei moved inwardly, myofilaments dissolved and disappeared mildly under the sarcolemma, there were scattered melting lesions within muscle fibers, the numbers of glycogen granules and mitochondria increased, mild hyperplasia and expansion of sarcoplasmic reticulum were seen, and a small number of muscle fibers had necrosis.</p><p><b>CONCLUSIONS</b>Weakness of both lower extremities remains the main reason for PMD patients seeing the doctor. CK is the main laboratory indicator for diagnosis of PMD. PMD is mainly manifested as myogenic damage in the early stage and may be accompanied by neurogenic damage in the late stage, according to the EMG examination. With a high misdiagnosis rate, PMD may be misdiagnosed as many other diseases. Pathological examination under light microscope and electron microscope is the main means for confirming a PMD diagnosis.</p>

Homing characteristics of graft cells in leukemia mice after allogenic bone marrow transplantation with different conditioning regimens / 中国实验血液学杂志

The aim of this study was to investigate the homing characteristics of bone marrow cells in leukemia mice after allogenic bone marrow transplantation with different conditioning regimens on the basis of a leukemia mouse model. Allogenic bone marrow transplantation was performed after three different kinds of conditioning regimen, including nonmyeloablative conditioning regimen (5 Gy (60)Co γ ray total body irradiation, A group), radiotherapeutic myeloablative conditioning regimen (9 Gy (60)Co γ ray total body irradiation, B group) and chemotherapeutic myeloablative conditioning regimen (large dose chemotherapy, C group). In the recipient mice, the nucleated cell number in peripheral blood, bone marrow and spleen was counted, the percentage of positive cells capable of connecting with FITC labeled anti-mouse H-2K(b) antibody was detected by flow cytometry and the homing ratio in bone marrow and spleen was calculated at 24, 48, 72, 96 h after bone marrow transplantation. The results showed that donor myeloid cells displayed homing and then mobilization (going out of home) in group A; homing, mobilization, and rehoming in group B and C, and there was a little delay of homing in the spleen in group C. In bone marrow, the homing efficiency of A group was the highest in early period and the lowest [(0.90 ± 0.09)%] in the fourth day with the mobilization of myeloid cells (P < 0.05), and the homing efficiency of B and C groups was lower in the early period and the highest [(2.17 ± 0.26)%, B group] in the fourth day with the rehoming of myeloid cells (P < 0.05). In spleen, the homing efficiency was similar to that in bone marrow and there still was a little delay in C group. It is concluded that the homing ratio is high in the early period and decrease obviously in 72 h after bone marrow of leukemia mice treated with nonmyeloablative conditioning regimen. The homing ratio is low in the early period and increases obviously in 72 h after bone marrow of leukemia mice treated with radio-or chemotherapeutic myeloablative conditioning regimens. The homing ratio does not obviously change between the early period and 72 h after bone marrow of leukemia mice treated with chemotherapeutic myeloablative conditioning regimen, and lies between group A and B.

A comparative study on psychological and behavioral development in children aged 1 to 3 years fostered by grandparents and by parents / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the differences of psychological and behavioral development between children aged 1 to 3 years fostered by grandparents and those by parents.</p><p><b>METHODS</b>Psychological and behavioral development of 443 children aged 1 to 3 years fostered by their grandparents and of aged-matched 443 children fostered by their parents were assessed with DST, an intellectual developmental screening test developed by Pediatric Hospital of Fudan University in Shanghai.</p><p><b>RESULTS</b>The abilities of social adaptation and intelligence development in children fostered by their grandparents were obviously retarded as compared with those in children fostered by their parents.</p><p><b>CONCLUSIONS</b>There are shortcomings in psychological and behavioral development in children aged 1 to 3 years fostered by grandparents.</p>

Clinicopathologic characteristics of breast cancer with basal-like immunophenotype / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic characteristics of basal-like immunophenotype breast cancer (BLBC).</p><p><b>METHODS</b>458 cases of female infiltrative breast cancer were studied using immunohistochemical staining with an antibody panel of ER, PR, HER2, Ki-67, CK5/6, CK14 and epidermal growth factor receptor (EGFR) and were classified basing on the immunophenotypes. The clinicopathologic characteristics were compared with other immunophenotypes of breast cancer. 228 of 458 cases of breast cancer were followed up.</p><p><b>RESULTS</b>46 cases of BLBC were screened out among the 458 breast cancers. And histological features of BLBC were analysed including the larger diameter of cancer foci (average 3.3 cm), appearance of squeezing phenomenon of neighboring cell borders (58.7%, 27/46), geography-like distribution of necrosis (52.2%, 24/46), central zone fibrosis (30.4%, 14/46) and lymphoplasmacytic infiltration at the margin and stroma (63.0%, 29/46). There were nuclear pleomorphism with numerous mitoses. The cancer cells were closely arranged, forming irregular solid architectures. There was a high expression (> 25%) of Ki-67 (43.5%, 20/46). CK5/6, CK14 and EGFR were positive in 58.7% (27/46), 43.5% (20/46) and 65.2% (30/46) respectively. 3-year survival rate of BLBC was 66.9%, lower than the luminal A breast cancer and similar to HER2 over-expression breast cancer.</p><p><b>CONCLUSIONS</b>The proportion of BLBC in the group of breast cancers is 10%. BLBC has its distinct histological and cytological features. Currently, it is still necessary to depend on immunophenotyping in making a BLBC diagnosis. BLBC is the one of breast cancer subtypes with the poorest prognosis.</p>

Construction of recombinant adenovirus vector containing human survivin gene and its expression in dendritic cells / 中国实验血液学杂志

The study was aimed to construct the recombinant adenovirus vectors containing human survivin gene, and to investigate their expression in transfected dendritic cells. Full length cDNA encoding survivin was obtained by PCR amplification from plasmid pcDNA3.0-survivin. The PCR product was restricted, and then inserted into pShuttle-CMV. The plasmids of pShuttle-CMV-survivin were linearized with PmeI, and the fragment containing survivin was ligated with pShuttle-CMV and transfected into E. coli BJ5183. After homologous recombination in bacteria, the extracted plasmid from the positive bacteria were linearized with PacI, transfected into HEK293 cells with liposome Lipofectamine 2000. Then, the harvested adenovirus supernatants were transfected into dendritic cells. The results showed that the recombinant adenovirus-survivin was constructed successfully and its titer was about 2.65 x 10(9) pfu/ml. The expression of survivin in transfected dendritic cells was confirmed by Western blot analysis. It is concluded that the recombinant adenovirus vector containing human survivin was constructed successfully, which may provide preliminary laboratory evidence for anti-leukemia immunotherapy.