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Objective To evaluate the value of fast-track surgery (FTS) in gynecological laparoscopy during the perioperative period. Methods The clinical data of 234 patients who underwent gynecological laparoscopy from January to December in 2018 were collected. Of note, 117 patients received a routine nursing method (control group), and 117 patients received the FTS nursing method (observation group). Results Compared with the control group, FTS group had a quicker postoperative bowel function recovery, earlier to get out of bed, and a lower anxiety level (P < 0.05); The incidence of postoperative nausea and vomiting of two groups had no statistical difference. Conclusions The application of FTS in gynecological laparoscopy is safe, effective, and worth promoting.

Mycoplasma infection is the most prevalent contamination in cell culture. Analysis of cell culture in laboratories from different countries shows that mycoplasma contamination ranges from 15% to 80% and, in some cases, even reaches 100% (Chernov et al., 2014). Whilst mycoplasma infection is not visible to the naked eye in cell culture, the consequences of mycoplasma contamination have been shown to induce a number of cellular changes, for example, increased resistance to chemotherapeutic drugs. Therefore, any results obtained from tissue culture studies, in the presence of mycoplasma contamination, potentially render the data invalid (Kim et al., 2015; Gedye et al., 2016). As such, mycoplasmas are not harmless bystanders and cannot be ignored in in vitro studies.

Purpose This study aims to incorporate informative histogram indicator analyses and advanced multimodal MRI parameters to differentiate low-grade gliomas (LGGs) from high-grade gliomas (HGGs) and to explore the features associated with patients' survival. Patients and methods A total of 120 patients with pathologically confirmed LGGs or HGGs receiving conventional and advanced MRI such as three-dimensional arterial spin labeling (3D-ASL), intravoxel incoherent motion-diffusion weighted imaging (IVIM-DWI), and dynamic contrast-enhanced MRI (DCE-MRI) were included. The mean and histogram indicators from advanced MRI were calculated from the entire tumor. The efficacies of a single indicator or multiple parameters were tested in distinguishing HGGs from LGGs and predicting patients' survival. Receiver operating characteristic (ROC) curve and multivariable stepwise logistic regression were used to evaluate the diagnostic efficacies. Leave-one-out cross-validation was further used to validate the accuracy of the parameter sets in glioma grading. Log-rank test using the Kaplan-Meier curve was utilized to predict patients' survival. Results Overall, parameters from DCE-MRI performed better than those from 3D-ASL or IVIM-DWI in both glioma grading and survival prediction. The histogram metrics of Ve were demonstrated to have higher accuracies (the accuracies for Extended Tofts_Ve mean and Extended Tofts_Ve median were 68.33% and 71.67%, respectively, while those for the Incremental_Ve mean and Incremental_Ve 75th were 68.33% and 72.50%, respectively) in grading LGGs from HGGs. The combination of Tofts_Ve histogram metrics was the one with the highest accuracy (81.67%) and area under ROC curve (AUC = 0.840). On the other hand, Patlak_Ktrans 95th (AUC = 0.9265) and Extended Tofts_Ve 95th (AUC = 0.9154) performed better than their corresponding means (Patlak_Ktrans mean: AUC = 0.9118 and Extended Tofts_Ve mean: AUC = 0.9044) in predicting patients' overall survival (OS) at 18-month follow-up. Conclusion DCE-MRI-derived histogram features from the entire tumor were promising metrics for glioma grading and OS prediction. Combining single modal histogram features improved glioma grading. Trial registration NCT02622620.

BACKGROUND Data about application of related haploidentical allogeneic hematopoietic stem cell transplantation (haplo-HSCT) on patients with high-risk or intermediate-risk acute myeloid leukemia (AML) in the first complete remission (CR1) are lacking. In this study, we report the outcomes of using unmanipulated haploidentical allogeneic peripheral blood stem cell transplantation (haplo-PBSCT) as post-remission therapy for patients with high-risk or intermediate-risk AML in CR1. MATERIAL AND METHODS From January 2008 to July 2016, 33 patients diagnosed as high-risk or intermediate-risk AML in CR1 undergoing haplo-PBSCT in our institution were enrolled for analysis. The cumulative incidence of platelet and neutrophil recovery, the occurrence of acute graft-versus-host-disease (GVHD) and chronic GVHD, relapse and non-relapse mortality were assessed. Patients' survival rates were estimated using the Kaplan-Meier method. RESULTS The cumulative incidence of grade 2-4 acute GVHD, overall and extensive chronic GVHD was 18.2%, 9.1%, and 6.1%, respectively. 2-year probability of relapse was 9.1%. Disease-free survival and overall survival at 2 years were 72.7% and 75.8%, respectively. CONCLUSIONS Our results showed that unmanipulated haploidentical transplantation with G-CSF primed PBSC alone as a graft source could be an acceptable alternative post-remission treatment for high-risk or intermediate-risk AML patients in CR1 lacking a matched donor.

To explore the correlation between the of regulatory T cells, Th17 cells and the prognosis of children with aplastic anemia.The clinical data 13 children with aplastic anemia (AA) treated by antithymocyte globulin (ATG) combined with Cyclosporine in Beijing Children's Hospital, Capital Medical University from January 2017 to January 2018 were analyized retrospectively. The changes of T cell and Th17 cell expression level in peripheral blood of AA children before and after IST for 6 and 12 month were compared and analyzed. The SPSS 19.0 statistical package was used for data analysis.Compared with the pre-IST, the expression level of Treg cells decreased at 6 months of IST, the difference was statistically significant (P＜0.05); however the expression level of Th17 cells did not show significant difference as compared with that pre-IST. The expression level of Treg and Th17 cells at 12 months of IST was lower than that pre-IST (P＜0.01), compared with the level pre-IST, the ratio of Treg cells/Th17 cell at 6 months and 12 months of IST did not show a singificand difference.Treg cells and Th17 cells in peripheral blood of AA children decrease after IST, which suggests that the change of regulatory T cells and Th17 cells correlate with the clinical outcome of children with aplastic anemia.调节性T细胞及Th17细胞的变化与儿童再生障碍性贫血临床预后的相关性.探讨Treg细胞及Th17细胞的变化与儿童再生障碍性贫血临床预后的相关性.回顾性分析2017年1月至2018年1月在首都医科大学附属北京儿童医院应用抗胸腺细胞球蛋白(ATG)联合环孢素免疫抑制治疗(IST)的13 例再生障碍性贫血(AA)患儿的临床资料。对比分析IST前和IST后6、12个月AA患儿外周血中Treg细胞及Th17细胞表达水平的变化, 采用 SPSS 19.0 统计软件进行数据分析.与IST前对比, IST后6个月Treg细胞表达水平明显降低, 差异具有统计学意义(P＜0.01)；而Th17细胞表达水平与IST前相比无统计学差异。与IST前对比, IST后12个月Treg和Th17细胞表达水平均明显降低(P＜0.01), 但IST后6个月、IST后12个月 Treg细胞/Th17细胞比值无明显变化.IST有效的AA患儿外周血Treg细胞、Th17细胞表达减少, 调节性T细胞及Th17细胞的变化与儿童AA的临床预后有相关性.

Background The medical imaging to differentiate World Health Organization (WHO) grade II (ODG2) from III (ODG3) oligodendrogliomas still remains a challenge. We investigated whether combination of machine leaning with radiomics from conventional T1 contrast-enhanced (T1 CE) and fluid attenuated inversion recovery (FLAIR) magnetic resonance imaging (MRI) offered superior efficacy. Methods Thirty-six patients with histologically confirmed ODGs underwent T1 CE and 33 of them underwent FLAIR MR examination before any intervention from January 2015 to July 2017 were retrospectively recruited in the current study. The volume of interest (VOI) covering the whole tumor enhancement were manually drawn on the T1 CE and FLAIR slice by slice using ITK-SNAP and a total of 1072 features were extracted from the VOI using 3-D slicer software. Random forest (RF) algorithm was applied to differentiate ODG2 from ODG3 and the efficacy was tested with 5-fold cross validation. The diagnostic efficacy of radiomics-based machine learning and radiologist’s assessment were also compared. Results Nineteen ODG2 and 17 ODG3 were included in this study and ODG3 tended to present with prominent necrosis and nodular/ring-like enhancement (P Conclusions Machine-learning based on radiomics of T1 CE and FLAIR offered superior efficacy to that of radiologists in differentiating ODG2 from ODG3.

BACKGROUND There is currently little information on haploidentical hematopoietic cell transplantation (haplo-HCT) for T-lymphoblastic lymphoma (T-LBL). Data about peripheral blood stem cells (PBSC) as a reliable graft source for T-LBL treatment are lacking. MATERIAL AND METHODS T-LBL patients who underwent T cell-replete haploidentical peripheral blood hematopoietic cell transplantation (haplo-PBHCT) from July 2007 to January 2017 were retrospectively evaluated. RESULTS A total of 25 patients (age ≥15 years) with median age of 24 (range 15-51) years were enrolled. The median number of CD34+ cells infused was 5.0 (1.6-14.4) 106/kg. Sustained myeloid engraftment with full donor chimerism was achieved in all patients. The cumulative incidence of grades 2 to 4 acute graft-versus-host disease (GVHD) at day 100 was 24%. Two-year extensive chronic GVHD cumulative incidence was 20%. The 3-year overall survival rate for all patients was 70%. The median survival time of the complete remission (CR) group was better than that of the non-CR group (not reached vs. 9 m) (P

// Zi-Yi Zhao 1, * , Sha-Sha Zhao 2, * , Meng Ren 2, * , Zi-Ling Liu 2 , Zhi Li 2 and Lei Yang 2 1 Department of Orthopaedics, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China 2 Cancer Center, The First Hospital of Jilin University, Changchun, China * Co-first authors Correspondence to: Lei Yang, email: dryangleilei@163.com Keywords: malignant pleural mesothelioma, hyperthermic intrathoracic chemotherapy, meta-analysis Received: June 30, 2017 Accepted: August 27, 2017 Published: October 19, 2017 ABSTRACT Surgery-based multimodality therapies have been used to control the malignant effusion and its recurrence in malignant pleural mesothelioma (MPM). Hyperthermic intrathoracic chemotherapy (HITHOC) has been used in the treatment of malignant pleural mesothelioma, but the results were controversial. The aim of the current study was, therefore, to conduct a systematic review and meta-analysis on the effect of HITHOC on MPM therapy. After thorough searching of online databases, total 21 articles were included into qualitative systematic review and 5 of them were used to conduct qualitative meta-analysis. It was found that most of HITHOC was used in combination of surgical resection including extrapleural pneumonectomy or pleurectomy/decortication. Patients who received HITHOC had significantly longer median survival length compared to the patients without HITHOC (Hedges’s g = 0.384 ± 0.105, 95% CI: 0.178~0.591, P < 0.001). In addition, HITHOC as palliative therapy was favored (Hedges’s g = 0.591 ± 0.201, 95% CI: 0.196~0.967, P < 0.001) in terms of recurrence free interval. The findings of the current study suggested that HITHOC is one of the safe and effective therapies in prolonging patients’ median survival time and extending recurrence free interval.

// Shasha Zhao 1, 2 , Zhenyang Gu 1 , Li Wang 1, 3 , Lixun Guan 1 , Feiyan Wang 1 , Nan Yang 1 , Lan Luo 1 , Zhe Gao 1 , Yingwei Song 4 , Lili Wang 1 , Daihong Liu 1 and Chunji Gao 1 1 Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China 2 Medical School, Nankai University, Tianjin 300071, China 3 Department of Hematology and Oncology, Laoshan Branch, No. 401 Hospital of Chinese PLA, Qingdao 266101, China 4 Department of Blood Transfusion, Chinese PLA General Hospital, Beijing 100853, China Correspondence to: Chunji Gao, email: gaochunji@medmail.com.cn Keywords: granulocyte colony-stimulating factor, CD4 + T cells, lymphocyte function-associated antigen-1, Lck, ZAP-70 Received: July 26, 2016 Accepted: May 06, 2017 Published: May 25, 2017 ABSTRACT In this study, we showed that G-CSF mobilization increased the frequency of T cells, specifically CD3 + CD4 + T cells. G-CSF mobilization decreased the secretion of inflammatory cytokines of CD4 + T cells through the LFA-1/ICAM-1 signaling pathway, whereas it did not alter the TH1/TH2 ratio. We found that G-CSF mobilization inhibited LFA-1-mediated CD4 + T cell polarization and motility. In vitro , G-CSF stimulation also attenuated the polarization and adhesiveness of CD4 + T cells through the LFA-1/ICAM-1 interaction. Further investigation revealed that G-CSF mobilization suppressed LFA-1 signaling by down-regulating Lck and ZAP-70 expression in CD4 + T cells, similar results was also confirmed by in-vitro studies. These findings suggested that G-CSF directly suppressed LFA-1-mediated CD4 + T cell functions through the down-regulation of Lck and ZAP-70. The immunosuppressive effect of G-CSF mobilization deepened our understanding about peripheral blood hematopoietic stem cell transplantation. LFA-1/ICMA-1 pathway may become a potential target for graft-versus-host disease prophylaxis.

The T790M mutational basis of treatment failure, following treatment via alteration of the epidermal growth factor receptor (EGFR) pathway, is a well-known anomaly in patients with non-small cell lung cancer (NSCLC). The T790M mutation activates the kinase domain, causing tyrosine kinase inhibitors, such as gefitinib, to elicit little or no response. To overcome this acquired resistance in NSCLC cells, the present study utilized a structure-based drug designing method to identify a novel lead compound. An in-house traditional Chinese medicinal compound database was used and following initial virtual screening, pre-absorption, distribution, metabolism and excretion/Tox and automated docking analyses, nardosinon was selected as the most appropriate candidate for further analysis. Two NSCLC cell lines, PC9GR4 and H2347, were used to test nardosinon and the results were compared with gefitinib. Results from an initial cell death assay revealed that nardosinon was able to induce cell death in NSCLC cells with and without the T790M mutation. These findings suggest that nardosinon may be an effective pharmacological compound for NSCLC treatment, including T790M EGFR mutant NSCLC cells.

Central nervous system lymphoma (CNSL) presents diagnostic and prognostic challenges. The aim of this meta-analysis was to evaluate the diagnostic and prognostic value of interleukin (IL)-10 in cerebrospinal fluid (CSF) for CNSL comprehensively. PubMed and Cochrane Library databases were searched through September 2016. Four studies with 212 CNSL patients and 262 control patients were included. The pooled sensitivity and specificity of CSF IL-10 for diagnosing CNSL were 81% (95% CI: 66-91%) and 97% (95% CI: 83-100%), respectively. The summary receiver operating characteristic (SROC) curve indicated that the area under the curve was 0.95 (0.93-0.97). The ROC curve based on extracted individual data showed that the optimal cutoff value was 6.88 pg/ml. Moreover, elevated CSF IL-10 was found to be associated with shorter progression-free survival (hazard ratio: 2.89, 95% CI: 1.13-7.41, p = .027). In conclusion, our meta-analysis showed that CSF IL-10 is an effective diagnostic and prognostic biomarker for CNSL.

To explore the clinical pathological features of the patients with diffuse large B cell lymphoma (DLBCL) and their prognostic factors.The prognosis of the clinical pathological features and their influence on prognosis of 177 patients diagnosed as DLBCL at the first visit from January 2013 to May 2017 in our hospital were analyzed retrospectively.The univariate analysis showed that overall survival (OS) and progression-free survival (PFS) were associated with later Ann Arbor stage (Ⅲ-Ⅳ) ( P＜0.01, P＜0.05), high performance status (ECOG score 2-4) (P＜0.01, P＜0.05), extranodal involvement ＞1 (P＜0.01, P＜0.05), elevated LDH level (P＜0.01, P＜0.05). B symptom (P＜0.05) and elevated β2-MG level (P＜0.05) also influenced OS. COX multivariate analysis showed that the elevated β2-MG level (P＜0.05) and later stage (Ⅲ-Ⅳ) (P＜0.05) have an independent influence on OS, later stage (Ⅲ-Ⅳ) (P＜0.05) also independently influenced PFS. The patients with high aaIPI score (2-3) and bone marrow involvement before treatment had poor OS (P＜0.01, P＜0.01) and PFS (P＜0.05, P＜0.01).Elevated β2-MG level can independently influence OS, and later stage (Ⅲ-Ⅳ) can independently influence both OS and PFS. High aaIPI score (2-3) and bone marrow involvement before treatment have an inferior influence on OS and PFS.年轻弥漫大B细胞淋巴瘤患者的临床病理特征与预后分析.探讨弥漫大B细胞淋巴瘤（diffuse large B cell lymphoma，DLBCL）患者的临床病理特征及其对预后评估作用.选取本院2013年1月至2017年5月177例确诊为弥漫大B细胞淋巴瘤的初治患者，回顾性分析其临床病理特征及其对预后的影响.单因素预后分析发现，分期Ⅲ-Ⅳ期（P=0.004，P=0.029）、ECOG评分2-4分（P=0.003，P=0.042）、结外受累＞1处（P=0.000，P=0.031）、异常升高的LDH值（P=0.000，P=0.046）为影响患者OS与PFS的危险因素；B症状（P=0.020）、异常升高的β2-MG值（P=0.020）仅影响患者OS。COX多因素分析显示，异常升高的β2-MG值（P=0.044）及分期（P=0.046）为影响患者OS的独立危险因素，分期（P=0.029）是影响患者PFS的独立因素。年龄调整的IPI（aaIPI）评分2-3分的患者与初治骨髓受累的患者OS（P=0.000，P=0.000）与PFS（P=0.013，P=0.000）更差.异常升高的β2-MG值是影响弥漫大B细胞淋巴瘤患者OS的独立因素，分期是影响患者OS与PFS的独立因素。aaIPI评分2-3分患者与初治骨髓受累的患者OS与PFS更差。.

Abstract (236 words) Background: The medical imaging differentiation of World Health Organization (WHO) grade II (ODG2) from III (ODG3) oligodendrogliomas remains a challenge. We investigate whether combination of machine leaning with radiomics from conventional T1 contrast-enhanced (T1CE) magnetic resonance imaging (MRI) can offer improved efficacy. Methods: Thirty-six patients with histologically confirmed ODGs who underwent the T1CE MR examination before any intervention between January 2015 and July 2017 were recruited in the current study. The volume of interest (VOI) covering the whole tumor enhancement were manually drawn on the T1CE slice by slice using ITK-SNAP and a total of 1044 features were extracted from the VOI using Analysis-Kinetics software. Random forest (RF) algorithm and 5-fold cross validation were applied to differentiate ODG2 from ODG3. The diagnostic efficacies of radiomics-based machine learning and radiologist’s assessment were also compared. Results: Nineteen ODG2 and 17 ODG3 were included in the current study and ODG3 tended to present with prominent necrosis and nodular/ring-like enhancement (P < 0.05). The RF strategy with radiomics features produced the stable diagnostic efficiency, with an AUC, ACC, sensitivity, and specificity of 0.765, 0.763, 82.8% and 70.0%, respectively. The AUCs of radiologists 1, 2 and 3 were 0.700, 0.687, and 0.714, respectively. The efficacy of machine learning based on radiomics is comparable to that of radiologist. Conclusions: Machine-learning based on radiomics of T1CE offered comparable efficacy to that of radiologist on differentiating ODG2 from ODG3.

Background The incidence and radiological patterns of eosinophilic granuloma (EG) in China is not clear. We described the incidence, presentation, and imaging characteristics of Chinese EG patients in a tertiary hospital. Methods A retrospective chart review was performed from January 2004 to October 2017 at a single tertiary general hospital. Seventy-six patients were pathologically identified as EG. Besides, 60 patients with preoperative imaging diagnosis of “EG” were analyzed to reveal the radiological patterns and their diagnostic power. Results Fifty-three male and 23 female EG patients with a mean age of 18.1 ± 16.7 years (range 1–58 years) were retrospectively included. Significant differences were observed in gender (male to female = 2.3:1) and age (the highest incidence at the age of 0~5 years) for EG. EG predominantly involved the skeletal system: flat bones (31.43%) > irregular bones (24.76%) > long bones (22.86%) > other organs (20.95%). No obvious relationships between season, biochemical markers, and EG incidence were observed. The common presenting symptoms were pain followed with local mass, and most patients underwent surgical resection. Among 60 imagingly diagnosed “EG” patients from April 2009 to October 2017, only 22 were with histological confirmation. The correct diagnosis rates were 37.1% (13 out of 35), 16.7% (5 out of 30), and 22.2% (8 out of 36) for plain radiography, computed tomography (CT), and magnetic resonance imaging (MRI), respectively. Conclusions Chinese EG has a varied presentation, age distribution, and gender difference. EG diagnosis is still based on biopsy or histopathology instead of imaging techniques. Electronic supplementary material The online version of this article (10.1186/s13018-019-1158-1) contains supplementary material, which is available to authorized users.

BACKGROUND Post-transplant lymphoproliferative disorder (PTLD) is a rare complication following solid organ transplantation and allogeneic hematopoietic stem cell transplantation (Allo-HSCT), which gives rise to high mortality rates. MATERIAL AND METHODS This was a single-center retrospective analysis based on 27 patients who were diagnosed with PTLD following Allo-HSCT between January 1, 2007 and June 2018 at the Chinese PLA General Hospital. The purpose of this analysis was to investigate responses and prognostic factors of rituximab-based treatment. RESULTS Twenty-seven patients were treated with rituximab. Among them, 20 of 27 patients (74.07%) had a complete response, 2 of 27 patients (7.41%) had a partial response, 5 of 27 patients (18.52%) had no response, and 22 of 27 patients (81.48%) cleared Epstein-Barr virus (EBV) copies. There were no obvious side effects. The 1-year overall survival (OS) estimate was 46.8% (95% CI, 23.1-65.5%). Univariate analysis revealed that lower OS was correlated with Eastern Cooperative Oncology Group (ECOG) score standard (3-4), Epstein-Barr virus (EBV) viral load (≥10⁶ copies/mL), bacteria or fungal infection, and EBV reactivation were positive after treatment with 1 or 2 doses of rituximab (P

Background: Thymic epithelial tumors (TETs) are the most common primary thymus tumors, but neither the possible ethnical/regional differences in the incidence of TETs nor the inter-relationships among the clinical variables has been revealed in northwest China. Methods: A retrospective chart review was performed among pathologically confirmed TET patients from January 2004 to December 2015 in a tertiary general hospital of northwest China and the incidence, clinical features and the inter-relationships among clinical variables were analyzed. Results: A total of 603 pathologically confirmed TETs patients (age range, 5–78 years; 308 males) were enrolled and the most common lesion location was anterior mediastinum (98.5%), among them, 192 (31.8%) had myasthenia gravis (MG). Twenty-six (5.7%), 112 (24.6%), 83 (18.2%), 137 (30.1%), 74 (16.3%), and 23 (5.1%) patients fell into the World Health Organization (WHO) type A, AB, B1, B2, B3 and thymic carcinoma (TC), respectively. The incidence of TETs was slightly higher in the female population and the age group of 40–60 years old. In addition, MG predominantly coexisted with WHO types A–B3 TETs and the TETs with MG were smaller than those without MG. The correct diagnosis rates were 42.3% (77 out of 182), 61.1% (127 out of 208), 89.3% (250 out of 280) and 75.0% (3 out of 4) for chest X-ray, non-contrast computed tomography (CT), contrast CT scan and magnetic resonance imaging (MRI), respectively. Conclusions: Distinct gender and age differences exist in the incidence of TETs and the A–B3 TETs are closely related with MG. Contrast CT scan plays more important role in diagnosing TETs.

MicroRNA-137 (miR-137) was reported to be dysregulated in several human cancers. However, the function and mechanism of miR-137 in non-small cell lung cancer (NSCLC) is still unclear. In the current study, we explored the role of miR-137 in NSCLC progression. Using qRT-PCR, our data showed that miR-137 was significantly down-regulated in NSCLC tissues and cell lines. In vitro functional assay, we found that over-expression of miR-137 suppressed NSCLC cells proliferation, migration and invasion, indicating that miR-137 could act as a tumor suppressor in NSCLC progression. In addition, bone morphogenetic protein-7 (BMP7) was identified as a target of miR-137 in NSCLC cells, Luciferase reporter assay suggested that miR-137 directly targeted 3’-UTR of BMP7, and correlation analysis revealed that BMP7 inversely correlated with miR-137 in NSCLC tissues. Furthermore, Restoration of BMP7 remarkably reversed the tumor suppressive effects of miR-137 on NSCLC cell proliferation, migration, and invasion. Taken together, our findings suggested that miR-137/BMP7 axis could contribute to the progression of NSCLC, suggesting miR-137 as a potential therapeutic target for the treatment of NSCLC.

To explore the killing effect of CAR (CD138-CD28-CD3ζ)-NK cells on myeloma cells through construction of CAR(CD138-CD28-CD3)-NK cells.The antiCD138scFv-CD28-CD3 zeta plasmid pcDNA3.1 was constructed, which then together with 3 plasmid lentiviral packaging system were transfected into 293T cells, the virus was collected. Furthermore, in order to get the stably transfected cell line, the NK92MI cell line was infected by the virus, then the positive cells were screened by puromycin. The expression of the CARNK cells were verified by RT-PCR and Western blot. At last the ability of secreting cytokine CD107a was detected by flow cytometry, and the statistical analysis was carried out to verify the anti-myeloma effect of CAR-NK cells.Gene fragment of the CAR(antiCD138scFv-CD28-CD3ζ) was constructed successfully by gene engineering technique in vitro, and the gene sequence was verified to be correct by sequencing. By virus packaging technology, the virus expressing the protein of the CAR was obtained. PCR and Western blot verified the expression of CAR fusion protein on the sufurce of NK cells. The cell killing experiment confirmed that the CAR-NK cells possessed the ability to secrete cytokine CD107a superior to control cells and showed the obvious killing effect on multiple myeloma cells.The CAR can be constructed in vitro, and express on NK92 cells. The CAR-NK cells can kill the multiple myeloma cells expressing CD138 antigen, thereby plays an antimyeloma effect.

The auditory system has the ability to adjust its structure and function as the environment changes, which is called auditory plasticity. In the auditory system, inferior colliculus (IC) is an important relay station, which accepts the ascending inputs from dorsal nuclei of lateral lemniscus (DNLL). The present study was aimed to investigate the role of the DNLL in the formation of the plasticity of IC neurons. Here, we used extracellular single unit recording and electrical stimulation to investigate the plasticity of IC neurons in Kunming mice. The results showed that after the cessation of 30-minute electrical stimulation on contralateral DNLL, 95% of the inhibited IC neurons and 86% of the facilitated IC neurons showed plastic changes. Moreover, 1 h after the contralateral DNLL stimulation was stopped, the plastic changes in 74% of the inhibited IC neurons vanished, but still were maintained in 26% of the inhibited IC neurons. These results suggest that the contralateral DNLL ascending input can induce plastic changes of IC neurons, and this kind of effect can be maintained for a certain period of time, which is beneficial to enhance the sound intensity sensitivity of IC neurons.

To establish the animal model of luciferase-transfected A20 murine B cell lymphoma, so as to provide experimental tools to explore the effect of graft versus tumor.Luciferase- labeled A20 cells were cloned with puromycin selection. Transfected A20 cells and CA20 cell line stably expressing luciferase gene was successfully obtained. The the bioluminicent imaging found that the tumor luminescence could be observed on day 8 of A20 cell inoculation, and the mean fluorescent intensity was increased along with the tumor growth. Compared with the BMT group, the survival rate and body weight of BMT+A20-LucA mouse transplantation model with luciferase- transfected A20 cells has been successfully established, thus laying a foundation for investigation of graft-versus-tumor.

To investigate the effect of granulocyte-colony stimulating factor (G-CSF) in vitro stimulation on the distribution of lymphocyte subset in healthy human.Peripheral blood mononuclear cells (PBMNCs) were collected from 8 healthy volunteers by density gradient centrifugation on Ficoll-PaqueThe levels of T cells in G-CSF group and G-CSF+ConA group were both higher than that in the control group (P0.001, P0.05). However, there were not significantly different in B cells and NK cells levels among the 3 groups. Furthermore, analysis of the effect of G-CSF on T cell subsets indicated that the levels of CD4The levels of CD4

BACKGROUND The efficacy and safety of sirolimus (SIR)-based graft-versus-host disease (GVHD) prophylaxis in patients who were subjected to allogeneic hematopoietic stem cell transplantation (allo-HSCT) remain to be clarified; this meta-analysis was conducted to evaluate these factors. STUDY DESIGN AND METHODS Data from original research were obtained from PubMed, Embase, and Cochrane central register of controlled trials databases. Randomized controlled trials (RCTs) evaluating the efficacy of SIR-based prophylaxis in allo-HSCT were included. The risk ratio (RR), with a 95% confidence interval (CI), was used to pool data. The random effects model was used, irrespective of the presence or absence of heterogeneity. RESULTS Five RCTs were included in the meta-analysis. SIR was observed to significantly decrease the incidence of Grade II to IV acute GVHD (aGVHD; RR, 0.65; 95% CI, 0.47-0.89). However, the incidence of Grade III to IV aGVHD and chronic GVHD was not decreased (RR, 0.91; 95% CI, 0.59-1.40; RR, 1.04; 95% CI, 0.88-1.23, respectively). An analysis of the toxic effects of SIR revealed that SIR effected a significant increase in the incidence of sinusoidal obstructive syndrome (RR, 2.24; 95% CI, 1.26-4.01), while that of thrombotic microangiopathy was not significantly increased (RR, 2.48; 95% CI, 0.87-7.06). Moreover, SIR did not improve event-free survival and overall survival (RR, 0.97; 95% CI, 0.85-1.10; and RR, 0.92; 95% CI, 0.82-1.02, respectively). CONCLUSION This meta-analysis indicated that the SIR-based regimen is an effective and safe alternative prophylaxis strategy for GVHD.

BACKGROUND Acute graft-versus-host disease (aGVHD) remains the most common and challenging complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT). An important obstacle to the therapeutic effect of aGVHD is the inability to identify risk factors for an individual patient at the onset of symptoms. We performed a retrospective study with the aim of defining clinically meaningful pre-transplantation risk factors for grades II-IV aGVHD patients. MATERIAL AND METHODS To identify pre-transplantation risk factors for grades II-IV aGVHD after allo-HSCT, we performed a retrospective study in 292 patients who underwent allo-HSCT at our center from January 2010 to July 2015. RESULTS The cumulative incidence of grades II-IV aGVHD was 36.6±2.8%. The most common target organ of aGVHD was the skin (46.7%), followed by the gastrointestinal tract (29.9%). The risk factors we identified included HLA-mismatched related donors (odds ratio [OR], 3.21; 95% confidence interval [CI], 1.82-5.67) and conditioning regimens containing TBI but no ATG (OR, 2.66; 95% CI, 1.40-5.06). Simultaneously, transplantation with an identical sibling donor (OR, 0.31; 95% CI, 0.18-0.55) and the use of ATG in the conditioning regimen containing TBI (OR 0.37; 95% CI, 0.15-0.93) were two factors found to be associated with a decreased risk of grades II-IV aGVHD. CONCLUSIONS Our study suggests that pre-transplantation characteristics of donor and recipient play an important role in identifying patients at high risk for grades II-IV aGVHD, which provide a direction for the prevention and treatment of aGVHD in the future.

// Zhenyang Gu 1, 2, * , Li Wang 1, 3, * , Lei Yuan 1, * , Wenrong Huang 1 , Meng Li 1 , Lixun Guan 1 , Qingyi Wang 4 , Zhe Gao 1 , Shasha Zhao 1 , Lan Luo 1 , Feiyan Wang 1 , Nan Yang 1 , Daihong Liu 1 , Jon C. Aster 2 and Chunji Gao 1 1 Department of Hematology, Chinese People’s Liberation Army (PLA) General Hospital, Beijing, China 2 Department of Pathology, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA, USA 3 Department of Hematology and Oncology, Laoshan Branch, No. 401 Hospital of Chinese PLA, Qingdao, China 4 Center for Computational and Integrative Biology, Massachusetts General Hospital, Boston, MA, USA * These authors have contributed equally to this work Correspondence to: Chunji Gao, email: gaochunji@medmail.com.cn Jon C. Aster, email: jaster@rics.bwh.harvard.edu Keywords: post-transplant cyclophosphamide, haploidentical, hematopoietic cell transplantation, HLA-matched, similar outcomes Received: January 17, 2017 Accepted: June 02, 2017 Published: June 29, 2017 ABSTRACT Background: Outcomes of haploidentical hematopoietic cell transplantation (haplo-HCT) with post-transplant cyclophosphamide (PT-Cy) have greatly improved. It remains unknown whether haplo-HCT with PT-Cy was associated with poor outcomes when compared with HLA-matched HCT. To address this issue, we performed a meta-analysis to compare outcomes of haplo-HCT with PT-Cy with those of HLA-matched HCT. Methods: A systematic search for case-control studies were performed in PubMed, Embase and Cochrane Library databases. Using a random model, the risk ratios (RRs) and 95% confidence intervals (95% CI) were pooled for the final analysis. Results: Nine case-control studies including 2258 patients (827 patients in the haplo-HCT with PT-Cy group, 748 controls from HLA-matched related donors (MRD), and 683 controls from HLA-matched unrelated donors (MUD)) met the inclusion criteria. No differences were found between haplo-HCT with PT-Cy and HLA-matched HCT with regard to acute graft-versus-host-disease (GVHD), non-relapse mortality, relapse, progression free survival and overall survival. However, haplo-HCT with PT-Cy was found to be associated with a lower incidence of moderate to severe chronic GVHD (Haplo vs MRD: RR=0.54; 95% CI=0.39-0.75; Haplo vs MUD: RR=0.70; 95% CI=0.56-0.88). Conclusions: The results of this meta-analysis suggest that haplo-HCT with PT-Cy can achieve comparable outcomes with those of HLA-matched HCT. Haploidentical donors can be a feasible and valid alternative when conventional HLA-matched donors are unavailable.

Immune thrombocytopenia (ITP) is an autoimmune disease characterized by increased platelet destruction and impaired platelet production. In this study, we conducted a systematic review and meta-analysis to determine the efficacy and safety of thrombopoietin receptor agonists (TPO-RAs) in primary ITP patients. Thirteen randomized controlled trials were included in this study, the pooled results of which demonstrated that TPO-RAs significantly increased platelet response (R) and durable response (DR) rates [risk ratio (RR): 2.77, 95% confidence interval (CI): 2.01–3.82, P = 5.9 × 10−10; RR: 7.52, 95% CI: 3.94–14.35, P = 9.2 × 10−10; respectively] and that TPO-RAs significantly reduced the incidences of any or severe bleeding events (RR: 0.80, 95% CI: 0.67–0.95, P = 0.013; RR: 0.52, 95% CI: 0.27–0.99, P = 0.048; respectively). Moreover, our results indicated that there was a significant reduction in the proportion of patients needing rescue medications in the TPO-RA groups compared with the control groups (RR: 0.50, 95% CI: 0.42–0.59, P = 2.0 × 10−15) and that the rates of any or severe adverse events were similar between the TPO-RA and control regimens (RR: 1.01, 95% CI: 0.92–1.10; RR: 0.74, 95% CI: 0.54–1.01; respectively). These findings demonstrate that TPO-RAs are an effective and safe second-line treatment option for primary ITP patients.

To explore the effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) mobilization on S1P5 expression in T lymphocyte subsets of allo-HSCT donors.The peripheral blood was collected from 10 allo-hematopoietic stem cell transplantation (allo-HSCT) donors before and after mobilization with rhG-CSF for 4 days. The flow cytometry was used to detect S1P5 expression in T lymphocyte subsets.There was no S1P5 expression on the surface of T-lymphocytes both before and after rhG-CSF mobilization. After fixation with permeabilization agent, S1P5 expression could be detected in lymphocytes after rhG-CSF mobilization, which indicates S1P5 may be located in cells. Compared with level before rhG-CSF mobilization, S1P5 expression was significantly increased in T lymphocyte subsets after rhG-CSF mobilization, CD3(+)T cells (57.92±2.32)% vs (7.94±1.47)%(P0.05）, CD4(+)T cells (72.58±1.73)% vs （5.48±0.82）%(P0.05）, CD8(+)T cells（51.79±3.57）% vs （6.46±1.01）%（P0.05）,CD3-/CD56(+)NK cells（40.00±1.47）% vs（4.97±0.74）%（P0.05）. The up-regulated level of S1P5 expression in CD4(+)T cells was most high(P0.05).S1P5 expression significantly increases in T lymphocyte subsets after rhG-CSF mobilization, and the up-regulated level of S1P5 expression in CD4(+)T cells is the most high.

To investigate the expression profile of lncRNA NONHSAT040475 in peripheral blood leukocytes of healthy persons.The peripheral blood mononuclear cells were collected from 10 healthy volunteers, the CD3(+) T cells,CD19(+) B cells,CD56(+) NK cells and granulocytes were purified and sorted by flow cytometry. Then, the expression of lncRNA NONHSAT040475 in peripheral blood leukocyte subsets was detected by real-time quantitative PCR.the expression levels of lncRNA NONHSAT040475 were various in lymphocyte subsets, its expression in B lymphocytes was significantly higher, as compared with T lymphocytes, while the expressions of lncRNA NONHSAT040475 in NK cells and granulocytes were relative low（P0.01）.lncRNA NONHSAT040475 is widely expressed in human peripheral blood leukocytes, and mainly expressed in B lymphocytes, therefore, laying the foundation for further study of B lymphocyte-related diseases. This study has brought a new opportunity for diagnosing and illustrating the molecular mechanism of hematologic disorder or autoimmune disease.

// Li Wang 1, 2, * , Haiyan Zhang 3, * , Lixun Guan 1 , Shasha Zhao 1 , Zhenyang Gu 1 , Huaping Wei 1 , Zhe Gao 1 , Feiyan Wang 1 , Nan Yang 1 , Lan Luo 1 , Yonghui Li 1 , Lili Wang 1 , Daihong Liu 1 , Chunji Gao 1 1 Department of Hematology, Chinese People’s Liberation Army (PLA) General Hospital, Beijing, China 2 Department of Hematology and Oncology, Laoshan Branch, No. 401 Hospital of Chinese PLA, Qingdao, China 3 Department of Hematology, Linyi People’s Hospital, Linyi, China * These authors have contributed equally to this work Correspondence to: Chunji Gao, email: gaochunji@medmail.com.cn Keywords: mesenchymal stem cells, graft-versus-host disease, hematopoietic stem cell transplantation, meta-analysis, animal experimentation Received: April 30, 2016 Accepted: July 28, 2016 Published: August 12, 2016 ABSTRACT A meta-analysis of animal models was conducted to evaluate the prophylactic effects of mesenchymal stem cells (MSCs) on acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation. A total of 50 studies involving 1848 animals were included. The pooled results showed that MSCs significantly reduced aGVHD-associated mortality (risk ratio = 0.70, 95% confidence interval 0.62 to 0.79, P = 2.73×10 -9 ) and clinical scores (standardized mean difference = -3.60, 95% confidence interval -4.43 to -2.76, P = 3.61×10 -17 ). In addition, MSCs conferred robust favorable prophylactic effects on aGVHD across recipient species, MSC doses, and administration times, but not MSC sources. Our meta-analysis showed that MSCs significantly prevented mortality and alleviated the clinical manifestations of aGVHD in animal models. These data support further clinical trials aimed at evaluating the efficacy of using MSCs to prevent aGVHD.

C20H24I2N2O2, monoclinic, P21/c (no. 14), a = 17.5052(10) Å, b = 12.7846(7) Å, c = 9.4663(5) Å, β = 94.899(1)°, V = 2110.8(2) Å3, Z = 4, R gt(F) = 0.0321, wR ref(F 2) = 0.0996, T = 291 K.

The tungsten carbide catalysts supported on carbon nanotubes were prepared by the temperature-programmed reduction, and there were used as catalysts for the hydrogenation of cellulose. The optimum synthesis and reactive conditions were investigated. The results show that the WC loading was 40%, the catalyst calcination temperature was 773K, the reduction temperature was 1073K. The conversion rate of cellulose could reach 84.5%, and the yield of sorbitol was 45.9%. The WC/CNTs catalysts show high catalytic activities and higher selectivity towards sorbitol in crystal cellulose hydrogenation.

The study compares the effects of three preservation methods (ethanol, drying and freezing) and two extraction methods (commercial kit and manual) on the PCR success rate and genotyping errors with fecal DNA from Przewalski’s horse (Equus przewalskii). The interaction term between preservation and extraction was significant, and the highest amplification success rate (0.661) was obtained with samples stored in ethanol and extracted manually. The preservation method and time significantly impacted microsatellite amplification success rates. The method of preservation had an effect on the rate of allelic dropout (ADO), but the rate of false alleles (FA) varied nonsignificantly. Feces preserved in ethanol yielded the lowest ADO and FA, followed by drying and freezing. Our optimized preservation protocol probably have wide applicability to equids and rhinocerotids that produce large, high-fiber fecal pellets.

A HPLC method with UV detection was developed and validated for the simultaneous determination of rivanol and mifepristone in human plasma. Norethisterone was used as the internal standard. Separation was performed by a C18 reversed-phase column maintained at 20 °C. The mobile phase was a mixture of methanol–acetonitrile–0.05% sodium dodecylsulfonate in a 0.05 M phosphate buffer with the pH adjusted to 3.0 (30:30:40, v/v/v) at a flow rate of 0.8 ml/min. Dual wavelength mode was used, with mifepristone monitored at UV 302 nm, while rivanol and norethisterone at 272 nm. A reliable biological sample pre-treatment procedure by means of solid-phase extraction was used, which allowed to obtain good extraction efficiency (>93%) for both of the analytes and the internal standard. The calibration curves were both linear with the correlation coefficient r equal to 0.9999. For rivanol, the assay gave CV% values for precision always lower than 7.8% and mean accuracy values higher than 95.3%. As to mifepristone, precision was always lower than 10.1% and mean accuracy values were higher than 93.8%. The limit of detection for the assay of rivanol and mifepristone was 1.1 and 3 ng/ml, respectively. The method is simple, sensitive and accurate, and allow for simultaneous determination of nanogram levels of rivanol and mifepristone in human plasma. It could be applied to assess the plasma level of rivanol and mifepristone in women undergoing polypharmacy with the two drugs.

Flexible Pipe is an essential equipment in marine oil and gas development. It connects undersea resources and platforms. In the complex marine conditions, it suffers effects from wind, wave and flow. In order to adapt the complex conditions, there appears the spiral layer in flexible pipe. Because it has the special function in complex conditions, it is widely used in various types of flexible pipe structure. In order to research the function of spiral layer, we use finite element software to analyze and compare different models to summary function of spiral layer in flexible pipe.

To evaluate the efficacy of mesenchymal stem cells (MSC) in the prevention of graft versus host disease (GVHD) after hematopoietic stem cell transplantation (HSCT).Randomized controlled trials (RCT) were identified from PubMed (1950.1-2014.3), EMbase (1970.1-2014.3), Cochrane Central Register of Controlled Trials (CENTRAL, issue 4, 2014) of the Cochrane Library, China Biological Medicine (CBM, 1978.1-2014.3). References of retrieved articles were also identified. The quality of each RCT was evaluated by the Cochrane collaboration's tool for assessing the risk of bias. Data analysis was performed with Review Manager 5.1 to evaluate the efficacy of MSC in the prevention of GVHD after HSCT.A total of 3 English articles involving 117 patients were included. Meta-analysis indicated that MSC did not reduce the incidence of acute GVHD and chronic GVHD (RR:0.44, 95% CI: 0.08 to 2.51, P = 0.35; RR:0.85, 95% CI: 0.54 to 1.33, P = 0.47). However, MSC did not increase occurrence of relapse and cytomegalovirus infection (RR:1.52, 95% CI:0.63 to 3.68, P = 0.35;RR:1.05, 95% CI:0.72 to 1.53, P = 0.78). Finally, MSC did not improve overall survival rate of patients received HSCT (RR:1.06, 95% CI:0.79 to 1.43, P = 0.71).MSC may have a preventive effect on GVHD in patients undergoing HSCT. However, the evidence is weak due to the small sample sizes. Thus, a reliable conclusion about the preventive effect of MSC on GVHD at the moment has not been made, further larger, high quality, randomized and controlled trials are warranted.

MicroRNA-137 (miR-137) was reported to be dysregulated in several human cancers. However, the function and mechanism of miR-137 in non-small cell lung cancer (NSCLC) is still unclear. In the current study, we explored the role of miR-137 in NSCLC progression. Using qRT-PCR, our data showed that miR-137 was significantly down-regulated in NSCLC tissues and cell lines. In vitro functional assay, we found that over-expression of miR-137 suppressed NSCLC cells proliferation, migration and invasion, indicating that miR-137 could act as a tumor suppressor in NSCLC progression. In addition, bone morphogenetic protein-7 (BMP7) was identified as a target of miR-137 in NSCLC cells, Luciferase reporter assay suggested that miR-137 directly targeted 3'-UTR of BMP7, and correlation analysis revealed that BMP7 inversely correlated with miR-137 in NSCLC tissues. Furthermore, Restoration of BMP7 remarkably reversed the tumor suppressive effects of miR-137 on NSCLC cell proliferation, migration, and invasion. Taken together, our findings suggested that miR-137/BMP7 axis could contribute to the progression of NSCLC, suggesting miR-137 as a potential therapeutic target for the treatment of NSCLC.

Objectives Controversy exists regarding whether oral cryotherapy can prevent oral mucositis (OM) in patients with hematological malignancies undergoing hematopoietic stem cell transplantation (HSCT). The aim of the present meta-analysis was to evaluate the efficacy of oral cryotherapy for OM prevention in patients with hematological malignancies undergoing HSCT. Methods PubMed and the Cochrane Library were searched through October 2014. Randomized controlled trials (RCTs) comparing the effect of oral cryotherapy with no treatment or with other interventions for OM in patients undergoing HSCT were included. The primary outcomes were the incidence, severity, and duration of OM. The secondary outcomes included length of analgesic use, total parenteral nutrition (TPN) use, and length of hospital stay. Results Seven RCTs involving eight articles analyzing 458 patients were included. Oral cryotherapy significantly decreased the incidence of severe OM (RR = 0.52, 95% CI = 0.27 to 0.99) and OM severity (SMD = -2.07, 95% CI = -3.90 to -0.25). In addition, the duration of TPN use and the length of hospitalization were markedly reduced (SMD = -0.56, 95% CI = -0.92 to -0.19; SMD = -0.44, 95% CI = -0.76 to -0.13; respectively). However, the pooled results were uncertain for the duration of OM and analgesic use (SMD = -0.13, 95% CI = -0.41 to 0.15; SMD = -1.15, 95% CI = -2.57 to 0.27; respectively). Conclusions Oral cryotherapy is a readily applicable and cost-effective prophylaxis for OM in patients undergoing HSCT.

The adhesion and polarization of T lymphocytes involved in the adhesive interaction of lymphocyte function-associated antigen 1 (LFA-1) with its ligand intercellular adhesion molecule 1 (ICAM-1). This study was aimed to investigate the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) stimulation in vitro on the adhesion and polarization of CD4⁺ T cells of healthy human in peripheral blood. The peripheral blood mononuclear cells were collected from 12 healthy volunteers. The CD4⁺ T cells were sorted by miniMACS. The sorted CD4⁺ T cells were incubated with rhG-CSF for 24 h, then the adhesion and polarization of CD4⁺ T cells activated by stroma cell-derived factor -1α (SDF-1α) and ICAM-1 were detected by ELISA and inverted microscope. The results showed that the percentage of adhesion CD4⁺T cells in the experimental group (rhG-CSF acting on the healthy adult volunteers) (61.9 ± 5.9)% was lower than that in the control group (healthy adult volunteers without rhG-CSF stimulation) (68.3 ± 7.3)% (P0.05). The percentage of polarized CD4⁺T cells in the experimental group (24.3 ± 4.3)% was also lower than that in control group (47.1 ± 5.1)% (P0.05). It is concluded that the adhesion and polarization of CD4⁺T lymphocytes can be inhibited after rhG-CSF stimulation.

The polarization and migration of T lymphocytes involves the adhesive interaction of lymphocyte function-associated antigen 1(LFA-1) with its ligand intercellular adhesion molecule 1 (ICAM-1). This study was aimed to investigate the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) mobilization on the polarization and migration of donor's CD4(+) T cells in peripheral blood. The peripheral blood mononuclear cells were collected from 10 healthy volunteers and 10 donors on the fifth day of mobilization with rhG-CSF. And the CD4(+)T cells were purified by miniMACS. The polarization and migration of CD4(+) T cells activated by stroma cell-derived factor -1α (SDF-1α) and ICAM-1 were detected by using inverted and confocal microscopes respectively. The results showed that the percentage of polarized CD4(+)T cells from donors(32.42 ± 4.91)% was lower than that from healthy controls(56.55 ± 5.35)% (P0.01), the migration velocity of CD4(+)T cells from donors (7.06 ± 1.44 µm/min) was also lower than that of healthy controls(9.05 ± 1.91 µm/min)(P0.01). It is concluded that the polarization and migration of CD4(+)T lymphocytes is impaired after rhG-CSF mobilization.

A total of 15 polymorphic microsatellite loci were isolated from forest musk deer (Moschus berezovskii). These loci showed high levels of allelic diversity, presenting 6–14 alleles per locus in a group with 31 individuals. The mean observed and expected heterozygosities were 0.82 and 0.68, respectively. The average polymorphic information content value, cumulative discrimination power, overall probability of exclusion with both unknown parents or with only unknown sire for these markers were 0.78, 1.0, 0.9999 and 1.0, respectively, indicating that this suit of microsatellite loci would be a powerful tool for the genetic determination of the forest musk deer.

Przewalski's horses have been imported from the western zoos to China since 1985. Yet the genetic diversity in China's populations has not been studied, thus lacking of such knowledge inevitably affects this population's management. The aim of this study was to assess genetic diversity in Chinese population of Przewalski's horses via mitochondrial DNA (mtDNA) control region and pedigree analysis.Two captive and one reintroduced populations were examined based on mitochondrial DNA control region variation via fecal sampling from 2010 to 2012, together with pedigree analysis.Amplification success rates of fecal mtDNA were as high as 96.2% (93.8%-100%), and were higher for sample in winter than in summer and autumn. Two haplotypes were identified and shared among three populations, but the proportion of individuals with each haplotype varied among the three populations (F(ST) = 0.10874, p = 0.00978). Haplotype diversity in the released population (0.153) was much lower than that in the two captive populations (0.4011 and 0.4966), in accordance with the direction of increase in probability of identity at the dam lines.Future concerns in Przewalski's horse population management should emphasize on strict reproduction control to minimize inbreeding in captivity, followed by long-term genetic diversity guidelines and non-invasive monitoring in the reintroduction programmes.

To construct the prokaryotic expression plasmid pEGX-6P-1-SAK-HC, express it in E.coli, and identify its biological activity.The fusion gene (SAK-HC) was obtained by overlap-extension PCR and then inserted into prokaryotic soluble pEGX-6P-1 vector with GST tag to construct expression plasmid (pEGX-6P-1-SAK-HC). GST-SAK-HC was expressed by E.coli B834 (DE3) under the induction of IPTG and purified by Glutathion-Sepharose 4B (GST) affinity chromatography and negative-ion exchange column (DEAE) chromatography. PreScission protease was used to remove the GST tag. The purity of the fusion protein was analyzed by SDS-PAGE and the fibrinolytic activity of SAK-HC in vitro was characterized by soluble fibrin plate method.PCR, sequencing and restriction enzyme digestion analysis demonstrated that the recombinant plasmid was constructed successfully. The fusion protein was expressed in E.coli B834 (DE3), M(r); being 36 000 as shown by SDS-PAGE. After purified by GST affinity and DEAE chromatography, SAK-HC fusion protein of high purity was obtained from the cell supernantants. In vitro experiments showed that the fibrinolytic activity of the recombinant SAK-HC was about 9.4×10();4 IU/mg.The SAK-HC fusion protein we obtained was successfully expressed in E.coli and exhibited a fibrinolytic activity as high as the urokinase standard, which offers a base for the identification of immunogenicity of the fusion protein.