MiR-203a-5p Inhibits Multiple Myeloma Cell Proliferation and Cell Cycle Progression via Targeting JAG1 / 中国实验血液学杂志

OBJECTIVE@#To investigate the biological function of miR-203a-5p and the underlying mechanism in multiple myeloma (MM).@*METHODS@#Three miRNA expression profiles (GSE16558, GSE24371 and GSE17498) were downloaded from the GEO database. The three miRNA expression profiles contained 131 MM samples and 17 normal plasmacyte samples. The robust rank aggregation (RRA) method was used to identify the differentially expressed miRNAs between MM and normal plasmacytes. In order to carry out cytological experiments, MM cell line with stable over-expression of miR-203a-5p was constructed with lentivirus. Expression levels of miR-203a-5p in MM cells were quantified by qRT-PCR. The effects of miR-203a-5p on MM cells were investigated using assays of cell viability and cell cycle. Cell proliferation was measured using the Cell Counting kit (CCK)8 assay. The percentage of cells in each cell cycle was measured with a FACSCalibur system. Xenograft tumor models were established to evaluate the role of miR-203a-5p in tumorigenesis in vivo . To elucidate the underlying molecular mechanisms of miR-203a-5p in mediating cell proliferation inhibition and cell cycle arrest in MM, we used TargetScan and miRanda to predict the candidate targets of miR-203a-5p. The potential target of miR-203a-5p in MM cells was explored using the luciferase reporter assay, qRT-PCR, and Western blot.@*RESULTS@#An integrated analysis of three MM miRNA expression datasets showed that the levels of miR-203a-5p in MM were notably downregulated compared with those in normal plasmacytes. Accordingly, the relative expression levels of miR-203a-5p were decreased in MM cell lines. In addition, overexpression of miR-203a-5p inhibited the proliferation and cell cycle progression of RPMI8226 and U266 cells. In vivo experiments demonstrated that upregulation of miR-203a-5p expression could significantly inhibit the tumorigenesis of subcutaneous myeloma xenografts in nude mice. Mechanistic investigation led to the identification of Jagged 1 (JAG1) as a novel and direct downstream target of miR-203a-5p. Interestingly, the reintroduction of JAG1 abrogated miR-203a-5p-induced MM cell growth inhibition and cell cycle arrest.@*CONCLUSION@#Our data demonstrate that miR-203a-5p inhibits cell proliferation and cell cycle progression in MM cells by targeting JAG1, supporting the utility of miR-203a-5p as a novel and potential therapeutic agent for miRNA-based MM therapy.

Short-Term Effect of Modified Kaijie Huatantang on Chronic Rhinosinusitis with Nasal Polyp Due to Qi Stagnation Phlegm Syndrome / 中国实验方剂学杂志

Objective:To observe the short-term effect of modified Kaijie Huatantang on chronic rhinosinusitis with nasal polyp due to qi stagnation phlegm syndrome. Method:A total of 90 cases were divided into control group and observation group，with 45 cases in each group. The control group was given mometasone furoate，and the observation group was given modified Kaijie Huatantang after nasal endoscopy surgery for 4 weeks. After treatment and follow-up for 1 years，the sino-nasal outcome test-20（SNOT-20），Lund-Kennedy and traditional Chinese medicine（TCM） syndrome were observed. The serum and nasal secretions tumor necrosis factor-alpha（TNF-<italic>α</italic>），interleukin（IL）-1<italic>β</italic>，IL-8，IL-17，eosinophilic cationic protein（ECP） and immunoglobulin E（IgE） were detected before and after treatment. The safety，clinical efficacy after treatment and follow-up for 1 years were compared between two groups. Result:After treatment and follow-up for 1 years，the total control rates were 97.7%，93.0% in observation group， which were higher than 87.8%，75.6% in control group（<italic>P</italic><0.05）. Compared with the control group after treatment and follow-up for 1 years，SNOT-20，Lund-Kennedy and TCM syndrome scores in the observation group decreased in the same period（<italic>P</italic><0.05）. Compared with the control group after treatment，the serum and nasal secretions TNF-<italic>α</italic>，IL-1<italic>β</italic>，IL-8，IL-17，ECP and IgE in the observation group were significantly decreased（<italic>P</italic><0.05）. The incidence of postoperative complications was 2.3% in the observation group， which was lower than 17.1% in the control group（<italic>P</italic><0.05）. The incidence of adverse reactions was 4.7% in the observation group， which was lower than 41.5% in the control group（<italic>P</italic><0.05）. Conclusion:Modified Kaijie Huatantang can significantly improve the short-term clinical efficacy of patients with chronic rhinosinusitis and nasal polyp due to Qi stagnation phlegm obstruction，with a low incidence of adverse reactions.

Effect of Modified Shuzhong Huatantang Replace Hormone on Children with Chronic Rhinosinusitis and Adenoid Hypertrophy due to Spleen Deficiency and Phlegm Obstruction After Operation / 中国实验方剂学杂志

Objective:To observe the effect of modified Shuzhong Huatantang replace hormone on children with rhinosinusitis and adenoid hypertrophy due to spleen deficiency and phlegm obstruction after operation. Method:Eighty cases were randomly divided into control group and observation group，40 cases in each group. After nasal endoscopy，the control group was given mometasone furoate，and the observation group was given modified Shuzhong Huatantang for 6 week. The nasal situation Lund-Kennedy assessment scal（Lund-Kennedy），adenoid thickness /nasopharyngeal cavity width （A/N），TCM syndrome were observed for before treatment，6，24，and 48 weeks after operation. The contents of immunoglobulin E（IgE），eosinophilic cationic protein（ECP），eosinophilic granulocyte（EOS），tumor necrosis factor-alpha（TNF-<italic>α</italic>），interleukin-1<italic>β</italic>（IL-1<italic>β</italic>），interleukin-17（IL-17） in serum and nasal secretions were detected before and 48 weeks after operation. The clinical efficacy，complications，recurrence 48 weeks after operation were compared between the two groups. Result:Three cases of abscission in the control group and one case in the observation group during the study period. The total control rate was 94.9% in the observation group higher than that 75.7% in the control group six weeks after the operation（<italic>χ</italic>²=6.972，<italic>P</italic><0.05）. The recurrence rate was 2.6% in the observation group lower than that 18.9% in the control group 48 weeks after operation（<italic>χ</italic>²=4.137，<italic>P</italic><0.05）. To compared with the control group at 6，24 and 48 weeks after operation，Lund-Kennedy，A/N，TCM syndromes in the observation group decrease in the same period （<italic>P</italic><0.05）. To compared with the control group 48 weeks after operation，The TNF-<italic>α</italic>，IL-1<italic>β，</italic>IL-17，IgE，ECP，EOS in serum and nasal secretions in the observation group were reduced （<italic>P</italic><0.05）. The incidence of adverse reactions was 2.7% in the observation group lower than that 29.7% in the control group. Conclusion:Modified Shuzhong Huatantang can significantly improve the postoperative clinical symptoms with chronic rhinosinusitis and adenoid hypertrophy due to spleen deficiency and phlegm obstruction，and the recurrence rate is lower.

TOSO interacts with SYK and enhances BCR pathway activation in chronic lymphocytic leukemia / 中华医学杂志(英文版)

BACKGROUND@#TOSO, also named Fas inhibitory molecule 3 (FAIM3), has recently been identified as an immunoglobulin M (IgM) Fc receptor (FcμR). Previous studies have shown that TOSO is specifically over-expressed in chronic lymphocytic leukemia (CLL). However, the functions of TOSO in CLL remain unknown. The B-cell receptor (BCR) signaling pathway has been reported to be constitutively activated in CLL. Here, we aimed to investigate the functions of TOSO in the BCR signaling pathway and the pathogenesis of CLL.@*METHODS@#We over-expressed TOSO in B-cell lymphoma cell lines (Granta-519 and Z138) by lentiviral transduction and knocked down TOSO by siRNA in primary CLL cells. The over-expression and knockdown of TOSO were confirmed at the RNA level by polymerase chain reaction and protein level by Western blotting. Co-immunoprecipitation with TOSO antibody followed by liquid chromatography coupled with tandem mass spectrometry (IP/LCMS) was used to identify TOSO interacting proteins. Western blotting was performed to detect the activation status of BCR signaling pathways as well as B-cell lymphoma 2 (BCL-2). Flow cytometry was used to examine the apoptosis of TOSO-over-expressing B lymphoma cell lines and TOSO-down-regulated CLL cells via the staining of Annexin V and 7-AAD. One-way analyses of variance were used for intergroup comparisons, while independent samples t tests were used for two-sample comparisons.@*RESULTS@#From IP/LCMS, we identified spleen tyrosine kinase (SYK) as a crucial candidate of TOSO-interacting protein and confirmed it by co-immunoprecipitation. After stimulation with anti-IgM, TOSO over-expression increased the phosphorylation of SYK, and subsequently activated the BCR signaling pathway, which could be reversed by a SYK inhibitor. TOSO knockdown in primary CLL cells resulted in reduced SYK phosphorylation as well as attenuated BCR signaling pathway. The apoptosis rates of the Granta-519 and Z138 cells expressing TOSO were (8.46 ± 2.90)% and (4.20 ± 1.21)%, respectively, significantly lower than the rates of the control groups, which were (25.20 ± 4.60)% and (19.72 ± 1.10)%, respectively (P < 0.05 for both). The apoptosis rate was reduced after knocking down TOSO in the primary CLL cells. In addition, we also found that TOSO down-regulation in primary cells from CLL patients led to decreased expression of BCL-2 as well as lower apoptosis, and vice versa in the cell line.@*CONCLUSIONS@#TOSO might be involved in the pathogenesis of CLL by interacting with SYK, enhancing the BCR signaling pathway, and inducing apoptosis resistance.

Effect of Modified Huading Loulutang on Postoperative Recovery of Chronic Rhinosinusitis with Secretory Otitis Media Due to Toxic Heat Syndromes / 中国实验方剂学杂志

Objective:To observe the efficacy of modified Huading Loulutang on postoperative recovery of chronic rhinosinusitis with secretory otitis media due to toxic heat syndromes and the effect of eosinophil cationic protein (ECP), immunoglobulin E (IgE) and intercellular adhesion molecule-1 (ICAM-1) in serum, nasal secretion and otitis media. Method:According to the random number table method, 90 cases were divided into control group and observation group, with 45 cases in each group. All patients were given ceftriaxone sodium + hydroxymezoline after nasal endoscopy. In addition, control group was given Biyuan Tongqiao granule, while observation group was given modified Huading Loulutang for 6 weeks. Lund-Kennedy nasal assessment scale, traditional Chinese medicine symptoms and threshold of hearing at different frequencies were observed in two groups before and after treatment and during 6-week and 18-week follow-up visits. The levels of ECP, IgE and ICAM-1 in serum, nasal and ear secretions were detected before and after treatment. Clinical symptoms, adverse reactions and 12-month recurrence rate were compared. Result:Total control rate was 97.7% (43/44, 95%CI [95.6，98.6]) in observation group, which was higher than 78.6% (33/42, 95%CI [72.4，83.5]) in control group (χ2=6.946, P<0.05). During the 12-month follow-up visit, the recurrence rate was 4.5% (2/44, 95%CI [3.7，5.8]) in observation group, which was lower than 19.0% (8/42, 95%CI[16.8，21.3]) in control group (P<0.05). After treatment, Lund-Kennedy and traditional Chinese medicine symptoms score during 6-week and 18-week follow-up visits in observation group were significantly lower than those in control group (P<0.05). Hearing threshold in observation group at different frequencies was significantly lower than that in control group (P<0.05). ECP, IgE and ICAM-1 levels in serum, nasal and auricular secretions in observation group were significantly lower than those in control group (P<0.05). The incidence of adverse reactions was 6.8% (3/44, 95%CI [5.3，8.1]) in observation group, which was lower than 28.6% (12/42,95%CI [25.3，30.2]) in control group (P<0.05). Conclusion:Modified Huading Loulutang can significantly improve postoperative clinical symptoms of chronic rhinosinusitis and otitis media due to toxic heat syndromes with secretion, with a low recurrence rate and fewer complications.

Effect of Wenfei Zhiliudan in Treating Lung Qi Deficiency Cold Type Allergic Rhinitis and Its Mechanism / 中国实验方剂学杂志

Objective: To observe the efficacy and safety of Wenfei Zhiliudan in treatment of lung Qi deficiency cold type allergic rhinitis. Method: A total of 120 cases were randomly divided into control group and observation group, with 60 cases in each group. The control group was given desloratadine + momethasone furoate, while observation group was given Wenfei Zhiliudan. A course of treatment was 2 weeks. The rhino conjunctivitis quality of life questionnaire (RQLQ), total nasal symptom score (TNSS) and traditional Chinese medicine (TCM) syndrome differentiation scale for lung Qi deficiency cold (TCM scale) in two groups before and after treatment were observed. The serum inflammatory factor[tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-33 (IL-33)]and immune indexes[immunoglobulin E (IgE), eosinophils (EOS), eotaxin (EOT)]were determined. The total effective rate and the incidence of adverse reactions were compared between two groups. Result: Five cases felt off during the study period. The total effective rate in observation group was 91.5%(54/59), which was higher than 82.5%(47/57) in control group (PPα, IFN-γ, IL-33) in observation group were significantly lower than those in control group (PPPPConclusion: Wenfei Zhiliudan can significantly alleviate the clinical symptoms, serum inflammatory factors and immune indicators of lung Qi deficiency cold type allergic rhinitis, with a lower incidence of adverse reactions.

Perfusion and cryopreservation of whole ovaries with vascular pedicles in common experimental animals / 解剖学报

Objective Using the perfusion needle which is authorized and special perfusion technology to perfuse the whole ovary with vascular pedicle of common animals, such as rabbits, Guinea pigs and rats, then comparative analysis of the adjustable perfusion needles whether can be applied to different animals, whether this kind of perfusion pressure and perfusion rate is appropriate for different sizes of ovaries, and using the classic cryopreservation protocol to freeze and thaw. Methods Collecting 12 ovaries respectively from 6 chinese sexual maturity rabbits, 6 Dunkan-Hartley Guinea pigs and 6 SD rats to do experiments. Results Through HE staining to count the normal proportion of primordial follicles in each section, the result of HE staining in three animals indicated that there was no statistical significance (P>0. 05), vascular injury was mainly in the upper (far away from ovary), there was no obvious damage in lower (close to ovary) in three animals' each group. Conclusion The experiment confirms that through adjusting straw needle size we can perfuse different size of animal organs and under the condition of the same perfusion pressure (60 mmHg) and rate (1 ml/min) it is suitable for rabbits' ovaries, Guinea pigs' ovaries and rats' ovaries at the same time.

Molecular mechanism of Mylabris for colorectal cancer based on integrative pharmacology / 中国中药杂志

At present, the disjointing situation is generally present between "systemic"and "local", "macro" and "micro", " process" and "activity evaluation" in the study of traditional Chinese medicine (TCM). An urgent task for the modernization of TCM is to establish new strategies and methods which can reflect the overall characteristics of TCM. The introduction of integrative pharmacology provided a feasible approach to solve the problem of the fragmentation of TCM. Internet-based computation platform method was adopted in this study to explore the active molecular mechanism of Mylabris in the treatment of colorectal cancer. Based on the analysis of the functional integration of internet-based computation platform V1.0 version software, the "core components-key target- main pathway" multidimensional network of Mylabris in treatment of colorectal cancer disease was constructed to explore the potential molecular mechanism of Mylabris in treatment of colorectal cancer from multiple perspectives. The results showed that Mylabris can treat the colorectal cancer and the mechanism might be associated with amino acid metabolism, NF-κB signaling pathway, immune system, endocrine system, nervous system, and chemokine signal transduction pathway, epithelial cell signaling in helicobacter pylori infection, T cell receptor signaling pathway, B cell receptor signaling pathway and so on.

Correlation analysis of "compound-target-pathway" of Yinchenhao decoction for cirrhosis based on computed prediction / 中国中药杂志

Chinese medicine prescription is the main form and means to treat diseases in traditional Chinese medicine (TCM). However, the disjointing situation is generally present between "systemic" and "local", "macro" and "micro", " process" and "activity evaluation" in the study of TCM at present. An urgent task for the modernization of TCM is to establish new strategies and methods which can reflect the overall characteristics of TCM. The introduction of integrative pharmacology provided a feasible approach to solve the problem of the fragmentation of TCM. Internet-based computation platform method was adopted in this study to explore the active molecular mechanism of Yinchenhao decoction in the treatment of cirrhosis. Based on the analysis of the functional integration of Internet-based Computation Platform V1.0 version software, the "core components-key target- main pathway" multidimensional network of Yinchenhao decoction in treatment of cirrhosis disease was constructed to explore the potential molecular mechanism of Yinchenhao decoction in treatment of cirrhosis from multiple perspectives. The molecular mechanism analysis of Yinchenhao decoction showed that Yinchenhao decoction can achieve the therapeutic effect on cirrhosis and the mechanism might be associated with oxidative phosphorylation, energy metabolism, circulatory system, glycerophospholipid metabolis, lipid metabolism and other pathways. Yinchenhao decoction in treatment of cirrhosis may be associated with energy metabolism and lipid metabolism.

Effect of ginsenoside Rg3 on intimal proliferation and apoptosis of vascular smooth muscle cells after vascular injury in rats / 中草药

Objective: To investigate the effects of ginsenoside Rg3 on intimal proliferation and apoptosis of vascular smooth muscle cells after vascular injury in rats. Methods: Forty Sprague Dawley rats were divided into four groups randomly including Sham operation group, model group, ginsenoside Rg3 low-dose group (5 mg/kg), and ginsenoside Rg3 high-dose group (10 mg/kg). The carotid artery intima injury model was established by inflation balloons. From the next day after modeling, the rats were treated with ginsenoside Rg3 by ig administation in ginsenoside Rg3 groups, and rats in Sham operation group and model group were administered with same amount of normal saline. The injured common carotid arteries were harvested after 14 d and morphological changes of injured arteries were observed by HE staining. Terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) was used to detect the apoptosis of smooth muscle cells; Western blotting and qRT-PCR were used to detect the expression of Fas and anti-apoptotic gene Bcl-2. Results: Compared with the Sham operation group, the vascular neointima of rats in model group was significantly thicker, and the ratio of intima/media area and the apoptosis rate of neointimal hyperplasia were significantly increased. The expression of apoptosis-related gene Fas and anti-apoptotic gene Bcl-2 was significantly increased. Compared with model group, ginsenoside Rg3 (5 and 10 mg/kg) significantly alleviated vascular intimal hyperplasia, the intima/media area ratio and the expression of Bcl-2 was significantly decreased, and the apoptosis rate of smooth muscle cells in the neointimal area and the expression of Fas in injured vessels were significantly increased. Conclusion: Ginsenoside Rg3 can reduce the vascular neointimal hyperplasia induced by balloon injury with the possible underlying mechanism to promote the apoptosis of smooth muscle cells.

Effect of miR-21 over-expression on proliferation, migration and diffe-rentiation of c-Kit+ cardiac stem cells / 中国病理生理杂志

AIM:To explore the effect of microRNA (miR)-21 on proliferation, migration and differentiation abilities of c-Kit+ cardiac stem cells (CSCs). METHODS:c-Kit+ CSCs were cultured and selected by the methods of en-zyme digestion and magnetic bead separation. miR-21 mimics (50 nmol/L) and mimics negative control ( MNC) were transfected into c-Kit+CSCs with Lipofectamine?2000. The cells was divided into 3 groups:control group:c-Kit+ CSCs without any pretreatment; MNC group:the cells were transfected with MNC for 48 h; mimics group:the cells were trans-fected with miR-21 mimics for 48 h. qPCR was used to assess the expression of miR-21 in each group. CCK-8 and EdU as-says were used to determine the cell proliferation. qPCR and immunofluorescence were used to detect the differentiation in each group. Scratch assay was adopted to explore the migration ability of the cells. RESULTS:The expression of c-Kit in the c-Kit+ CSCs were 90.8%, with 0.6% of CD45 and 0.5% of CD34. A significant increase in miR-21 expression was observed when the cells were transfected with miR-21 mimics for 48 h ( P<0.05). CCK-8 and EdU assays showed that miR-21 significantly increased cell proliferation as compared with MNC group and control group (P<0.05). No difference in the expression of Nkx2.5, CD31 and α-SMA at mRNA and protein levels was observed, and no difference of the migra-tion ability in 3 groups of the c-Kit+ CSCs was found. CONCLUSION:Over-expression of miR-21 significantly promotes the proliferation of c-Kit+ CSCs, without any effect on the cell migration and differentiation.

Effect of Ginsenoside Rg3 on NF-κBp65 and Its Related Inflammatory Factors after Carotid Balloon Injury in Rats / 中国药学杂志

OBJECTIVE: To study the effects of ginsenoside Rg3 on NF-κBp65 and its related inflammatory factors after carotid balloon injury in rats. METHODS: Sprague Dawley(SD) rats(n=40) were divided into 4 groups randomlysham operationgroup, model group, ginsenoside Rg3 5 mg·kg-1 and ginsenoside Rg3 10 mg·kg-1. 2.0 mm×12 mm Ballon catheters were used to establish the carotid artery intima injury model. On next day after modeling, all animals in model group were administered intragastrically with relative saline and different concentration of ginsenoside Rg3 for 14 d. After 14 d, the morphological changes of the injured arteries were observed by HE staining. The expression of NF-κBp65 in vascular tissue was detected by Western blot; Tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and IL-6 were detected by RT-PCR and Elisa. RESULTS: Compared with the sham-operation group, the neointimal in model group was significantly thicker(P<0.01). while the expression levels of IL-1β, IL-6, TNF-α and NF-κBp65 were increased in model group(P<0.01). The vascular intimal hyperplasia was alleviated distinctly(P<0.01) and the protein expression of NF-κBp65 in vascular tissue was significantly decreased(P<0.01) compared with model group. the expression level of IL-1β, IL-6 and TNF-α was significantly lower in intervention group(P<0.01). CONCLUSION: Ginsenoside Rg3 could reduce the vascular neointimal hyperplasia and inflammation induced by balloon injury, which may be related to its inhibitory role of the transcription factor NF-κB signaling pathway.

Efficacy and Safety of Bortezomib in Multiple Myeloma Patients with Hepatitis B: A Multicenter Retrospective Study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The efficacy and safety evidence of bortezomib in multiple myeloma (MM) patients with hepatitis B is vacant. This study aimed to investigate the efficacy and safety of bortezomib in MM patients with hepatitis B in China.</p><p><b>METHODS</b>From 2006 to 2011, 739 newly diagnosed MM patients were screened for serum hepatitis B virus (HBV) biomarkers. HBV-infected patients were followed for HBV reactivation by monitoring of serum alanine transaminase (ALT) and HBV DNA load. The pattern of HBV reactivation in relation to bortezomib was evaluated. Seven hundred thirty-nine MM patients were included in this study.</p><p><b>RESULTS</b>The prevalence of MM patients infected with HBV was 3.4% (n = 25), of which 17 cases were treated with bortezomib. Bortezomib had no significant influence on liver function (ALT before and after treatment: 36.69 ± 8.90 U/L vs. 11.31 ± 2.74 U/L, P = 0.19) and HBV DNA of MM patients with HBV (detectable HBV DNA percentage: 5.9% vs. 11.8%, P = 0.12).</p><p><b>CONCLUSIONS</b>Bortezomib can be used safely and effectively in MM patients with hepatitis B. HBV prophylaxis and surveillance are recommended during the MM treatment.</p>

Clinical significance of serum vascular endothelial growth factor in patients with multiple myeloma / 中国实验血液学杂志

This study was purposed to investigate the relation of serum vascular endothelial growth factor (VEGF) levels with clinical types and therapeutic efficacy of multiple myeloma (MM), and to analyze the significance of VEGF in MM. The levels of serum VEGF were detected by enzyme-linked immunosorbent assay (ELISA) technique in 76 patients with MM. The relationship between the serum VEGF levels with MM patients' age, stages, types, and efficacy were analyzed. The results showed that the patients who were less than 65 years old had higher serum VEGF levels than elder patients, however, the difference between them had no statistical significance (P > 0.05). The VEGF level was the highest in IgG type patients, and then in light chain type, lowest in IgA type, however there were no statistical differences between them (P > 0.05). Patients of DS stage III had higher VEGF level than that of stage II, and there was also no statistical difference (P > 0.25). Patients of ISS stage I had lower VEGF level than that of stage II and III, and it also showed no statistical difference (P > 0.05). After treatment, patients obtained complete remission (CR) or very good partial remission (VGPR) had decrease of serum VEGF level, however, patients obtained less than partial remission (PR) had increase of serum VEGF level. Patients were divided into two groups according serum VEGF level ( ≤ 150 ng/L), patients with high VEGF levels had short overall survival time, there was statistical difference (P = 0.03). It is concluded that the serum VEGF level of MM patients dose not relate with age, clinical stages and M protein types; however, there was a certain association between overall survival and serum VEGF level, and the later may be one of poor prognostic factors.

Different dose combinations of bortezomib and dexamethasone in the treatment of relapsed or refractory myeloma: an open-label, observational, multi-center study in China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Although previous clinical study revealed that bortezomib combined with dexamethasone had improved the outcomes of relapsed or refractory multiple myeloma (RRMM), the optimal dose combinations of bortezomib and dexamethasone remain unknown. This trial aimed to observe the efficacy and safety of different dose combinations of bortezomib and dexamethasone in the treatment of RRMM patients in China.</p><p><b>METHODS</b>A total of 168 patients with relapsed multiple myeloma (MM) who were refractory to at lest two prior treatments were enrolled in this multicenter, open-label, non-randomized, prospective clinical trial. Twenty patients received 1.3 mg/m(2) of bortezomib twice weekly for 2 weeks of a 3-week cycle for up to 8 cycles and oral or intravenous dexamethasone 20 mg on the day of and after each bortezomib dose (group 1); 66 patients received less than 1.3 mg/m(2) (0.7 - 1.0 mg/m(2)) of bortezomib and dexamethasone 20 mg on the same schedule (group 2); 37 patients received 1.3 mg/m(2)2 of bortezomib and dexamethasone 40 mg (group 3) and 45 patients received less than 1.3 mg/m(2) (0.7 - 1.0 mg/m(2)) of bortezomib and dexamethasone 40 mg (group 4). The response was evaluated according to the criteria of the European Group for Blood and Marrow Transplantation and confirmed by an independent review committee. Adverse events were graded according to the National Cancer Institute Common Toxicity Criteria, version 3.0.</p><p><b>RESULTS</b>The median age of groups 1 to 4 was 61, 62, 56, and 60 years, respectively. Most patients were in stages II/III of MM and the most common subtype was IgG. The rate of overall response to bortezomib and dexamethasone of group 1 to 4 was 72.2% (13/18), 73.8% (48/65), 78.8% (26/33) and 78.0% (32/41) (P = 0.91), including a complete response rate of 22.2% (4/18), 20.0% (13/65), 33.3% (11/33) and 29.3% (12/41) (P = 0.67), respectively. There was no statistical significance in time to progression and overall survival among these 4 groups (P > 0.05). The most commonly adverse events of any grade in the entire 4 groups were fatigue, gastrointestinal effects, peripheral neuropathy and thrombocytopenia, and there was no significance in the number of adverse events among the 4 groups (P > 0.05) except that peripheral neuropathy was reported more frequently in group 3 (36.3%) than in group 2 (13.8%, P < 0.05) and group 4 (14.6%, P < 0.05).</p><p><b>CONCLUSIONS</b>The combination of bortezomib and dexamethasone was associated with high responses in Chinese RRMM patients. No significant differences of efficacy were detected in different dose combinations of bortezomib and dexamethasone. Moreover, low dose of bortezomib reduced the incidence of peripheral neuropathy without affecting outcome in the treatment of patients with RRMM in China.</p>

Osteoblasts from patients with myelodysplastic syndrome express multiple cytokines and support hematopoietic progenitor cell survival in vitro / 中国实验血液学杂志

This study was aimed to investigate the biological characteristics of osteoblasts from patients with myelodysplastic syndrome (MDS) and their supportive capacity for hematopoiesis in vitro. A two-dimensional culture system was constructed by using osteoblasts derived from human marrow mesenchymal stem cells (MSC); MSCs were isolated from bone marrow of MDS patients and normal individuals and were cultured; the third passage of MSCs were induced into osteoblasts which were treated with mitomycin C and confluenced into a feeder layer. Ficolled bone marrow mononuclear cells were obtained from normal individuals and seeded into the two-dimensional culture system to culture in vitro without exogenous cytokines. By using colony-forming assay, the ability of the two-dimensional system to culture HPCs was observed. The cytokine expression of osteoblasts from MDS patient bone marrows in mRNA level was detected by RT-PCR and was compared with human osteoblast cell line hFOB1.19. The results showed that the osteoblasts from MDS patients could support short-term survival of GM-CFC in condition without exogenous cytokines, that is, osteoblasts played a crucial role in regulation of HPC growth. The results of RT-PCR clearly demonstrated that the osteoblast cell line hFOB1.19 expressed SCF, IL-6, SDF-1alpha, G-CSF and GM-CSF. The same expression patterns of above cytokines were also seen in osteoblasts derived from BM-MSCs of MDS patients and normal individuals, but these cells did not express GM-CSF. It is concluded that the biological characteristics of osteoblasts from bone marrow of MDS patients are generally not different from those of osteoblasts from normal bone marrow. Both of them can support GM -CFC to form colonies in vitro, it may be associated with expressing important related cytokines by osteoblasts.

Expression of human telomerase reverse transcriptase and survivin gene in patients with myelodysplastic syndrome / 中国实验血液学杂志

This study was aimed to investigate the expressions of human telomerase reverse transcriptase (hTERT) and survivin gene in patients with myelodysplastic syndrome (MDS), and to explore their relationship. The expression of hTERT mRNA in bone marrow mononuclear cells (BMMNCs) of 56 patients with MDS and 27 patients with iron deficiency anemia were detected by RT-PCR, the expressions of survivin gene in BMMNCs of 55 patients with MDS and 12 patients with iron deficiency anemia were detected by real-time RT-PCR. The results showed that the expression of hTERT significantly elevated in RA and RAEB patients, as compared with controls (p<0.005). With the disease alleviated, the expression of hTERT decreased and had no significant difference from the controls (p>0.25). There was no significant difference in expression of hTERT between low+int-1 risk group and int-2+high-risk group by IPSS (p>0.50). The expression of survivin gene significantly increased in RA and RAEB patients, as compared with controls (p<0.02, p<0.05). The expression of survivin gene in low+int-1 risk group by IPSS was significantly higher than that in the controls (p<0.02), and there was no significant difference in expression of survivin gene between int-2+high-risk group patients and the controls (p>0.10). It is concluded that the expressions of hTERT and survivin may play a critical role in escaping malignant clone of MDS from apoptosis and acquiring the ability to divide unlimitedly.

Biological characteristics of human fetal osteoblastic 1.19 cell line / 中国实验血液学杂志

This study was aimed to investigate the biological characteristics of osteoblasts and their hematopoietic supportive function by using human fetal osteoblastic cell line 1.19 (hFOBs) as a model. The pluripotency markers (Oct-4, Rex-1, hTERT) of hFOBs were analyzed by RT-PCR, the multilineage differentiation experiments were conducted in vitro. Flow cytometry (FCM) was used to identify the surface markers of hFOBs, and RT-PCR was used to analyze their hematopoietic cytokine expression in comparison with bone marrow mesenchymal stem cell (BM-MSC). The results showed that hFOBs expressed several ESC pluripotency markers including Oct-4 and Rex-1, except hTERT. Moreover, hFOBs could also undergo multilineage differentiation into the mesodermal lineages of adipocytic cell types in addition to its predetermined pathway, the mature osteoblast. Both hFOBs and BM-MSC expressed CD44, CD73 (SH3), CD105 (SH2) and CD90 (Thy1), and lack expression of CD34, CD45, or HLA-DR surface molecules. In addition, both hFOBs and BM-MSC expressed SCF, IL-6, and SDF-1alpha mRNA, but only hFOBs could express GM-CSF and G-CSF. It is concluded that human fetal osteoblastic cell line 1.19 may provide a good model to study the osteoblastic regulation role in hematopoiesis in vitro.

Anti-myloma activity of T cell activated by dentritic cells loading antigen of U266 cells / 中国实验血液学杂志

This study was aimed to investigate the effect of T cells activated by DCs loaded with whole antigens of U266 cells on the U266 cells survival in vitro. Peripheral blood mononuclear cells were isolated from healthy donor, and adherented on culture plate. Adherent cells were cultured in AIM-V serum-free medium or in RPMI 1640 medium contained 20% fetal bovine serum (FBS), supplemented with granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate killing rate of U266 cells by T cells activated by DCs loaded with whole antigen of U266 cells. The results showed that DCs derived from peripheral blood mononuclear cells cultured by AIM-V serum-free medium or RPMI 1640 medium containing FBS had similar immunophenotype. T cells activated by DCs loaded with whole antigen of U266 cells or mature DCs might kill U266 cells in a dose-dependent manner. It is concluded, DCs derived from peripheral blood mononuclear cells of healthy donor and loaded with whole antigen of U266 cells can induce anti-myeloma response of T cells in vitro.

Effects of uremia patients sera on bone marrow derived CFU-E proliferation / 中国基层医药

Objective To investigate the existence of the erythropoiesis inhibitors(?).Methods Twelve patients suffered from uremia with anemia were studied[5 males,and 7 females,(50?12)years].Methylcellulose culture technique was used to culture mice bone marrow cells.The sera from the uremia patients were added to CFU- E and BFU-E culture medium with final concentrations of 1,25%,2.5% and 5%,Mice bone marrow cells were ob- tained from the female Balb/c mice.In vitro CFU-E and BFU-E culture in the presence of sera from uremia patients was compared with that in the presence of normal human subjects with the use of normal mice bone marrows.Re- suits The effects of the sera from uremia patients on CFU-E and BFU-E colon growth were in a dose-dependent manner.The effect was correlated with the concentrations of the sera(P