Objective:To investigate the differentially expressed miRNAs in serum collected post operation and compared these miR-NAs with those collected pre-surgery among patients suffering from glioblastoma multiform (GBM) and undergoing regular clinical fol-low-up. These miRNAs may be potential biomarkers for the post-operative evaluation of patients with GBM. Methods:Forty-eight pa-tients with GBM and clinical pathological diagnosis were enrolled in this study. In the initial biomarker screening stage, total RNAs were extracted and subjected to Solexa sequencing to select miRNAs with significantly altered expression pre-and post-operation. Some of these differentially expressed miRNAs were chosen and verified through TaqMan probe-based qRT-PCR assay. A t-test was performed to determine the miRNAs that satisfied the two criteria, namely, fold change>2 and P<0.05. All of the patients were fol-lowed-up, and survival data were collected. The patients were then classified into two groups, namely, long-and short-survival groups, on the basis of the median of the miR-30e expression levels in the sera collected post-operation. Kaplan-Meier method and Log-rank test (SPSS version 19.0, IBM) were employed to determine the possible relationships between miR-30e expression levels in the sera collected post-operation and patients' overall survival. Results: Solexa revealed 63 differentially expressed miRNAs. Four miRNAs, namely, miR-26b, miR-30e, miR-129-3p, and miR-206, were selected on the basis of previous and present findings. These miRNAs were then verified in the RT-qPCR phase. Among these miRNAs, only miR-30e was significantly upregulated post-operation. The serum miR-30e expression level post-operation was not significantly associated with the overall survival of the patients. A low miR-30e expression level corresponded to prolonged survival. Conclusion:miR-30e was upregulated in the sera collected post-operation from patients with GBM. This miRNA may be negatively related to the tumor load of these patients. The miR-30e expression level in the serum col-lected post-surgery serum was not significantly associated with overall survival. Therefore, miR-30e may serve as a novel potential non-invasive biomarker for the post-operative evaluation of patients with GBM.

In this paper is presented an analysis of the mechanical effect of horizontal rotating bioreactor on cell culture. Getting the microgravity of the bioreactor and the shear stress on canine mesenchymal stem cells (cMSCs) with theoretic calculating model and differential equations, we have validated the density,growth rate and modality of cultured cell by scanning electron microscopy. The horizontal rotating bioreactor which we developed could create the mechanic environment of microgravity (K<8.38 X 10(-2))and low shear stress(r<1.62 dyn/cm2) in theory. The results of scanning electron microscopy indicated that the cells' growth-speed, quantity and modality in bioreactor were better than those of cells cultured in static 24-well plate. The mechanical environment of the rotating bioreactor is propitious for keeping better modality and more rapid proliferation of cMSCs. The rotating bioreactor is a novel approach and technique it is superior to static culture.
Animals ; Bioreactors ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; instrumentation ; methods ; Cell Proliferation ; Cells, Cultured ; Dogs ; Mechanotransduction, Cellular ; physiology ; Mesenchymal Stromal Cells ; cytology ; Rotation ; Tissue Engineering ; methods

Objective:To compare the differences in the delineation of the gross tumor volume (GTV) and lymph nodes of nasopharyngeal carcinoma (NPC) patients using computerized tomography (CT), magnetic resonance imaging (MRI), and 18F-fluorodeoxyglucose positron emission tomography/computed tomography ( 18F-FDG PET/CT), and to investigate the optimal standard uptake value (SUV; relative to the MRI-based delineation) for the automatic delineation of GTV using PET. Methods:A total of 53 NPC patients proposing to receive radiotherapy were selected for this study. The CT, MRI, and PET images of each patient were obtained before radiotherapy. Then GTV and positive lymph nodes were delineated on these three types of images. They were individually named GTV MRI, GTV CT, GTV PET2.5 (SUV=2.5), Lymph MRI, Lymph CT, and Lymph PET2.5 and compared. The GTV ∩2.5 (overlapped GTV) was obtained through the alignment of MRI and PET/CT images. Meanwhile, GTV was delineated on PET images using thresholds of SUV=4.0, 4.5, 5.0, and 5.6, obtaining GTV PET4.0, GTV PET4.5, GTV PET5.0, and GTV PET5.6. Then their volume and Dice similarity coefficients (DSCs) were compared. Results:Compared to GTV MRI, GTV CT decreased by 1.73% ( P>0.05) and GTV PET2.5 increased by 21.34% ( t=-3.52, P < 0.05) in the three types of images. The volume of Lymph PET2.5 was 1.61 and 1.87 times the volume of Lymph MRI and Lymph CT, respectively ( t=-4.12, -5.18; P< 0.05). The volume of high-SUV lymph nodes was 4.07 times the volume of lymph nodes with low SUVs or SUV=0 ( t=5.50, P< 0.05) on PET images. The DSC between GTV PET4.0and GTV MRI was 0.78 ± 0.27, which was lower than that between GTV PET2.5 and GTV MRI (0.84 ± 0.18). However, GTV PET4.0 approximated to GTV ∩2.5 ( P>0.05). Conclusions:Compared to CT and 18F-FDG PET/CT, MRI shows more accurate boundaries of GTV and lymph nodes. When 18F-FDG PET/CT was adopted to automatically delineate GTV, the GTV delineated using SUV=4.0 was closer to GTV MRI.

Objective To develop liver cancer-targeted nanoparticles(LA-CMGL)co-loaded with miR-145/camp-tothecin(CPT)and assess their targeting specificity,combined anti-tumor effects,and magnetic resonance imaging efficacy.Methods Laser scanning confocal microscopy and flow cytometry were utilized to evaluate the targeted uptake of lactobionic acid-modified and unmodified nanoparticles by HepG2 cells and HepaRG cells;quantitative polymerase chain reaction(qPCR)was employed to assess miR-145 content in tumor cells and tissues.The cytotox-icity of CPT,LA-CPT-LNPs and LA-CMGL on HepG2 cells were assessed using the CCK-8 assay.qPCR was also used to evaluate the effect of CPT,LA-CPT-LNPs and LA-CMGL on apoptosis of HepG2 cells.MRI was performed to measure the relaxation rate of LA-CMGL and evaluate its targeting imaging effect on liver cancer cells.Results The uptake rate of LA-CMGL by HepG2 cells surpassed that of CMGL significantly.The relative miR-145 content in liver cancer cells and mouse liver cancer tissues in the LA-CMGL group was markedly higher compared to free miR-145 and CMGL groups(P＜0.001).The apoptosis rate of HepG2 cells in the LA-CMGL group exceeded that in the CMGL group and CPT group(P＜0.01).At the same Gd3+concentration,the relaxation rate of LA-CMGL significantly surpassed that of Gd-DOTA,and the MRI signal of LA-CMGL in HepG2 cells markedly increased com-pared to CMGL and Gd-DOTA.Conclusion LA-CMGL exhibits promising liver cancer-targeted delivery,com-bined anti-tumor effects,and MRI liver cancer cell-targeted imaging,offering a novel avenue for combined drug/gene therapy for liver cancer.

Objective: To investigate the relationship between the erythrocyte membrane protein gene mutations and the clinical severity of hereditary spherocytosis (HS). Methods: Targeted sequencings were performed on 25 HS patients, correlation between HS mutations and patients’ clinical characteristics were evaluated. Results: A total of 25 HS patients were enrolled, including 13 males and 12 females with median age of 20 (4-55) years, including 9 compensatory hemolysis patients, 9 patients with mild anemia, 3 patients with moderate anemia and 4 patients with severe anemia. Of them, 18 patients (72%) harbored HS-related mutations, including ANK1 mutation in 6 cases, SLC4A1 mutation in 6 cases, SPTB mutation in 5 cases and 1 case with EPB41 mutation. Seven patients (28%) didn’t carry common HS mutations. SPTB and SLC4A1 mutations mainly affected male patients. There was no significant difference between the age of diagnosis (P=0.130) and HGB level (P=0.585) in patients with HS mutation and those without mutation, however, the EMA binding fluorescence intensity (P=0.015), AGLT50 (P=0.032) and EOF minimal hemolytic concentration (P=0.027) were significantly different in these two groups of HS patients. Conclusion To screen erythrocyte membrane protein coding gene mutations could favor the diagnosis of HS, and patients without mutations have mild clinical phenotype.

Objective To investigate the effect of Angelica polysaccharides on the balance of Th17/T-regulatory(Treg)cells and the expression of related inflammatory factor proteins in the bone marrow of mice with aplastic anemia.Methods 50 male BALB/c mice were randomly divided into a normal group,model group,cyclosporine A(CsA)group,and angelica polysaccharide low-(APS-L)and high-dose(APS-H)groups.A mouse model of aplastic anemia was prepared by combining irradiation with allogeneic lymphocyte infusion.After modeling,the mice were orally administered with corresponding drugs for 28 days.The general condition,weight changes,and spleen index of the mice were observed.Blood samples were taken to test for changes in basic hematological parameters of peripheral blood,and hematoxylin and eosin staining was used to evaluate the pathological changes in mouse bone marrow tissue.An ELISA method was applied to detect bone marrow TGF-β1.The expression levels of proteins such as IL-10,IL-17A,and IL-6 were analyzed,and flow cytometry was used in detecting the bone marrow Th17/Treg cell ratio.Results Compared with the normal group,the model group mice showed clumsiness during activity;lethargy;pale eyelids,lips,and ears;and reduced food and water intake.Both body weight and spleen index were significantly decreased.Peripheral red blood cell,white blood cell,and platelet counts and hemoglobin concentration were significantly reduced.A disordered bone marrow tissue structure and significantly reduced proliferation of hematopoietic tissue were seen.The expression of IL-17A and IL-6 proteins was significantly upregulated,and the expression of TGF-β1 and IL-10 protein was significantly downregulated.The proportion of Th17 cells in bone marrow significantly increased,the proportion of Treg cells significantly decreased,and the ratio of Th17/Treg cells significantly increased(P＜0.01).Compared with the model group,the general condition of the APS-L and APS-H groups improved,showing an increase in food and water intake;a significant increase in body weight and spleen index;a significant increase in peripheral red blood cells,white blood cells,platelets,and hemoglobin concentration(P＜0.05);an improvement in bone marrow tissue structure;and an increase in hematopoietic tissue proliferation(P＜0.05).The expression of IL-17A and IL-6 proteins was significantly downregulated.The expression of TGF-β1 and IL-10 protein was significantly upregulated(P＜0.05 or P＜0.01).The proportion of Th17 cells in bone marrow significantly decreased,the proportion of Treg cells significantly increased,and the ratio of Th17/Treg cells significantly decreased(P＜0.01).Conclusions Angelica polysaccharides can improve bone marrow hematopoietic function in aplastic anemia mice,and its mechanism of action may include the regulation of Th17/Treg cell balance.

Objective To observe the application effect of magnetic resonance imaging technology in evaluating the brain structure and function of patients with type 2 diabetic painful neuropathy(PDN).Methods Forty patients with type 2 diabetes mellitus hospitalized in our hospital were se-lected as the study objects,and were divided into diabetes mellitus(DM)group(n=12),peripheral neuropathy(DPN)group(n=14)and PDN group(n=14).General clinical biochemical indexes of three groups were analyzed.The structural brain and function of brain area in three groups were com-pared.Results Age,duration of diabetes,systolic blood pressure,diastolic blood pressure,fasting blood glucose(FBG),glycosylated hemoglobin(HbA1c),free fatty acid(FFA),albumin(ALB),creatinine(Cr),uric acid(UA),estimated glomerular filtration rate(eGFR),cystatin C(Cys-C),total cholesterol(TC),triglyceride(TG)of the three groups were compared,high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),triiodothyronine(T3),thyroxine(T4),thyroid stimulating hormone(TSH),thyroglobulin antibody(TGAb),thyroid peroxidase anti-body(TPO-Ab)and serum calcium(Ca)in the three groups showed no significant differences(P＞0.05).Compared with the DM group,the gray matter volume(GMV)in the DPN group was signifi-cantly decreased(P＜0.05).Compared with the DM group,amplitude of low frequency fluctuation(ALFF)and fractional amplitude of low frequency fluctuation(fALFF)of left medial superiorfrontal gyrus in the PDN group were significantly decreased(P＜0.05).Conclusion Abnormal GMV in the left angular gyrus in DPN patients may be associated with a higher risk of concomitant cognitive impairment.The decrease of ALFF in the right cerebellar vermis and fALFF in the left medial superior frontal gyrus in PDN patients may be related to the pathogenesis of pain.

Objective To observe the application effect of magnetic resonance imaging technology in evaluating the brain structure and function of patients with type 2 diabetic painful neuropathy(PDN).Methods Forty patients with type 2 diabetes mellitus hospitalized in our hospital were se-lected as the study objects,and were divided into diabetes mellitus(DM)group(n=12),peripheral neuropathy(DPN)group(n=14)and PDN group(n=14).General clinical biochemical indexes of three groups were analyzed.The structural brain and function of brain area in three groups were com-pared.Results Age,duration of diabetes,systolic blood pressure,diastolic blood pressure,fasting blood glucose(FBG),glycosylated hemoglobin(HbA1c),free fatty acid(FFA),albumin(ALB),creatinine(Cr),uric acid(UA),estimated glomerular filtration rate(eGFR),cystatin C(Cys-C),total cholesterol(TC),triglyceride(TG)of the three groups were compared,high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),triiodothyronine(T3),thyroxine(T4),thyroid stimulating hormone(TSH),thyroglobulin antibody(TGAb),thyroid peroxidase anti-body(TPO-Ab)and serum calcium(Ca)in the three groups showed no significant differences(P＞0.05).Compared with the DM group,the gray matter volume(GMV)in the DPN group was signifi-cantly decreased(P＜0.05).Compared with the DM group,amplitude of low frequency fluctuation(ALFF)and fractional amplitude of low frequency fluctuation(fALFF)of left medial superiorfrontal gyrus in the PDN group were significantly decreased(P＜0.05).Conclusion Abnormal GMV in the left angular gyrus in DPN patients may be associated with a higher risk of concomitant cognitive impairment.The decrease of ALFF in the right cerebellar vermis and fALFF in the left medial superior frontal gyrus in PDN patients may be related to the pathogenesis of pain.

Objective : To explore the association of time in range(TIR) and glucose management indicator ( GMI) with the risk of type 2 diabetic nephropathy (DN) ． Methods : The clinical data of 215 patients with type 2 diabetes mellitus (T2DM) were collected and analyzed．According to the results of estimated glomerular filtration rate (eGFR) and urinary albumin to creatinine ratio( UACR) ，they were divided into 117 patients with T2DM and 98 patients with DN．The clinical data，biochemical indicators and continuous glucose monitoring ( CGM) indicators of the two groups were compared．Logistic regression was used to analyze the influencing factors of DN risk．The predictive value of TIR and GMI on the risk of DN was evaluated by receiver operating characteristic (ROC) curve. Results: There were significant differences in age，duration of diabetes，systolic blood pressure，glycosylated hemoglobin ( HbA1c) ，fasting plasma glucose (FPG) ，2 hour postprandial plasma glucose (2hPG) ，creatinine( Cr) ，UACR, eGFR between the two groups(P＜0. 05) ．There were statistically significant differences between the two groups in the CGM indexes of GMI，mean absolute difference of mean of daily differences ( MODD) ，glucose above target range time(TAR) and TIR(P＜0. 05) ．The results of logistic regression analysis showed that TIR was a protective factor of DN．In the ROC curve analysis of TIR prediction DN，the area under the ROC curve was 0. 718 (95% CI = 0. 648 ～0. 789，P＜0. 001) ，and the Yoden index was 0. 38．At this time，the sensitivity was 66. 7% ，and the specificity was 71. 3%．In the ROC curve analysis of GMI prediction DN，the area under the ROC curve was 0. 701 (95% CI = 0. 629 ～0. 774，P＜0. 001) ，and the Yoden index was 0. 368．At this time，the sensitivity was 63. 3% ， and the specificity was 73. 5%． Conclusion Specifically，lower TIR and higher GMI increase the risk of DN.

To investigate the resistance mechanisms of tigecycline-non-susceptible Acinetobacter baumannii and for providing the evidence of the control of nosocomial infection and rational use of antibiotics. The minimum inhibitory concentrations (MICs) of 94 non repetitive tigecycline-non-susceptible A. baumannii from 20 hospitals in 12 cities of China were determined by agar dilution method and broth microdilution method. The molecular epidemiology was studied by Multilocus sequence typing (MLST) and eBURST software. PCR and sequencing techniques were used to analyze the resistance genes (blaOXA-40-like, blaOXA-58-like, blaOXA-23-like, blaOXA-51-like, blaNDM-1), ISAba1, and the mutation sites of adeR, adeS, and trm. The activity of polymyxin B and minocyclinem against tigecycline-non-susceptible A. baumannii were 100% and 25.5%, respectively. The sensitivities of other antibiotics were less than 3.5%, and the sensitivities of imipenem and meropenem totigecycline-nonsusceptible A. baumannii were only 1.1%. A total of 12 ST types were identified, including ST195 (45, 47.9%), ST208 (19, 20.2%) and ST457 (10, 10.6%). EBURST analysis found that 8 of the ST types belonged to the clone complex 92 (Clonal Complex 92, CC92). The blaOXA-23-like type carbapenem gene was identiefied in 93 strains (99% positive); and none of the strains contained the blaNDM-1 gene. The detection rates of adeR and adeS were 73.4% and 91.5% respectively and high frequency mutation sites were located in adeR (Asp26Asn) and adeS (Ala97Glu); The ISAba1 located upstream of the adeS gene was detected in 12 strains of A. baumannii, mainly from the northern region of China. The 240 nucleotide deletion of the trm gene caused a frameshift leading to a premature stop. So the tigecycline-non-susceptible A. baumannii showed high resistance against most antibiotics except polymyxin B. The deletion and mutation of adeR, adeS and trm were the main resistant mechanisms in tigecycline-non-susceptible A. baumannii in China.