Objective To investigate the role of Th17 cells in chronic myeloid leukemia (CML) pathogenesis. Methods 33 CML patients [15 newly diagnosed (ND)- and 18 chronic-phase (CP)- CML patients] and 15 healthy controls were enrolled. The percentage of Th17 cells in the peripheral blood (PB) of CML and controls were evaluated by flow cytometry. RORC mRNA expressions were examined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). The levels of IL-17 in PB of CML and controls were measured by enzyme-linked immunoadsorbent assay (ELISA). The relationships between Th17 cells and clinical characteristics in CML were analyzed. Results The percentage of Th17 cells in PB of ND-CML patients [(0.71±0.41) %] was significantly lower than that of CP-CML patients [(4.08±0.74) %, P<0.05] and healthy controls [(3.18±1.32) %, P<0.05]. The mRNA level of RORC in PB of ND-CML patients (0.043 3±0.040 5) was significantly decreased compared with that in CP-CML patients (0.086 1±0.052 3, P<0.05) and healthy controls (0.091 0±0.058 4, P<0.05). The levels of IL-17 in PB of ND-CML patients [(1.43±0.22) pg/ml] and CP-CML patients [(1.36±0.19) pg/ml] were slightly higher than that of healthy controls [(1.23±0.14) pg/ml, P< 0.05]. The percentage of Th17 cells had significantly negative correlations with white blood cell counts in PB or bcr-abl (IS). Conclusion Th17 cells may play an important role in CML pathogenesis, which has potential implication for immunotherapy of this malignancy.

Objective To update the knowledge on the sensitizing drugs and clinical features of drug eruption. Methods The clinical data on 138 patients hospitalized for drug eruption in the Department of Dermatology, Institute of Dermatology, Chinese Academy of Medical Sciences, from January 2005 to June 2007, were collected and retrospectively analyzed. Results Totally, 178 episodes of drug eruption were observed in these patients during the tested period. The major sensitizing drugs included antibacterial agents (31.46%), non-steroidal anti-inflammatory drugs (28.09%), traditional Chinese medicines (15.73%). Amoxicillin triggered 20 episodes of drug eruption and was the most common causative drug. Oral administration was the predominant sensitizing route of administration (54.17%). Of all the drug eruptions, 33.71% manifested by erythema multiforme, 28.09% by fixed drug eruption, 22.47% by exanthematous drug eruption. Severe types of drug eruption were mainly caused by traditional Chinese medicines and anti-gout drugs. Conclusions Antibacterial agents and non-steroidal anti-inflammatory drugs have become the major sensitizing drugs of drug eruption, especially amoxicillin. The frequency of traditional Chinese medicine-induced eruptions are increasing. Furthermore, caution is warranted for the drug eruption caused by oral administration.

Objective To investigate the effects of ulinastatin on the number of peripheral blood lymphocyte and the percentage of its subsets in patients with severe sepsis. Methods The scores of APECHE II and SOFA, the number of lymphocyte and the percentage of different subsets in these sepsis patients at different treatment time were measured. Results After treatment, the scores of APECHE II and SOFA of severe sepsis patients were decreased, the number of lymphocyte elevated and the percentages of different subset were corrected. Sepsis caused by Gram- positive pathogens had stronger suppression of peripheral blood lymphocyte and subsets compared with Gram - negative pathogens. Conclusion Patients with severe sepsis had less peripheral blood lymphocyte and abnormal subsets. Ulinastatin could help to correct such abnormality.

Sugarcane bagasse modified by polyethylenimine (PEI) and glutaraldehyde (GA) was used as a carrier to immobilize Clostridium acetobutylicum XY16 in the process of butanol production. The effects of chemically modified sugarcane bagasse on batch and repeat-batch fermentations were investigated. Batch fermentation was conducted with an addition of 10 g/L modified sugarcane bagasse and 60 g/L glucose, resulting in a high solvent concentration of 21.67 g/L and productivity of 0.60 g/(L x h) with the treatment of 4 g/L PEI and 1 g/L GA. Compared to the fermentations by free cells and immobilized cells on unmodified sugarcane bagasse, the productivity increased 130.8% and 66.7%, respectively. The fibrous-bed bioreactor also maintained a stable butanol production during repeat-batch fermentations, achieving a maximum productivity of 0.83 g/(L x h) with a high yield of 0.42 g/g.
Batch Cell Culture Techniques ; Bioreactors ; Butanols ; metabolism ; Cells, Immobilized ; Cellulose ; metabolism ; Clostridium acetobutylicum ; metabolism ; Fermentation ; Saccharum ; chemistry

This study examined the current changes of human ether-a-go-go-related gene (hERG) mutation derived from a LQT2 Chinese family with a highly penetrating phenotype. Mutation was identified and site-directed mutagenesis was performed to induce the mutation in wild-type (WT) hERG. WT hERG and mutated V535M were cloned and transiently expressed in HEK293 cells. At the 48th and 72nd h after transfection, membrane currents were recorded using whole cell patch-clamp procedures. An A>G transition at 1605 resulting in replacement of V535M was identified. Compared to WT, V535M mutation significantly decreased tail currents of hERG. At test potential of -40 mV after depolarizing at +50 mV, tail current densities were 83.35±7.06 pA/pF in WT and 50.38±7.74 pA/pF in V535M respectively (n=20, P<0.01). Gating kinetics of hERG revealed that V (1/2) of steady-state inactivation shifted to negative potential in the mutant (V (1/2,V535M): -61.81±1.7 mV vs. V (1/2, WT): -43.1±0.71 mV). The time constant of recovery from inactivation was markedly prolonged in the mutant compared to WT among test potentials. V535M hERG mutation demonstrated markedly decreased tail current densities, which suggests that V535M is a new loss-of-function mutation of hERG channel responsible for LQT2.

Objective To investigate the effects of exercise on expression of microtube associated protein-2 (MAP-2) and MAP-2 mRNA in neuronal cells after intracerebral hemorrhage (ICH) in rats. Methods Ninety-six male Sprague-Dawley rats (weighing 270 to 300 g) were divided into 3 groups, a trial group (ICH-induction and ex-ercise, n=32), a control group (ICH-induction only, n=32) and a sham-operated group (no ICH and no exercise, n=32). The brains of the rats were sampled at 7, 14, 21, and 28 days after the operation for establishing ICH mod-el. Another 64 rats were divided into 8 groups (6 h, 12 h, 24 h, 48 h, 72 h, 7 d after ICH, no ICH group and nor-mal group). MAP-2 and MAP-2 mRNA activity were measured by immunohistochemical methods and RT-PCR. The rats in the trial group began cage-running exercise 72 h after the operation. The others were reared in the standard ca-ges. Results ①MAP-2-positive cells appeared around the hematoma in the cortex. The number of MAP-2-positive cells was very small in the sham-operation group. There was an up-regulation of MAP-2 from 24 h to 7 d after ICH, and significant difference was found between the non-ICH group and the normal group. The expression of MAP-2 showed an up-regulation trend in the trial and control groups. There was a significant difference compared with the sham operated group, and the trial group had significantly higher expression levels than the control group. ②RT-PCR showed up-ragulation of MAP-2 mRNA 12 to 24 h after ICH. There was a significant difference between the no ICH and normal groups. The trial and control groups showed up-regulation of MAP-2 mRNA at 14 to 28 d after ICH, with was significantly different from that of the sham group, and the trial group had significantly higher levels than the con-trol group. Conclusion MAP-2 might participated in neural cells plasticity after ICH, and exercise training (cage-running) can up-regulate MAP-2 and MAP-2-mRNA expression.

Objective To investigate the possible mechanism of natural killer cells(NK cells)in immune dysfunction in sepsis by monitoring the phenotype and function of periphery NK cells in patients with sepsis. Methods A retrospective study was conducted. The patients with systemic inflammatory response syndrome(SIRS,n=59)or sepsis(n=65)admitted to Department of Critical Care Medicine of Anhui Provincial Hospital from August 2011 to August 2013 were enrolled. Blood samples were collected within 48 hours after intensive care unit(ICU)admission,the phenotype and function of periphery NK cells were determined by flow cytometry. Twenty-eight healthy people served as controls. Results The proportion and number of peripheral blood CD3-CD56+NK cells in SIRS and sepsis groups were normal,and no statistical difference was found when compared with those of the healthy control group〔cell proportion:0.102±0.019,0.102±0.108 vs. 0.106±0.018,F=0.018,P=0.982;cell number(×106/L):182.46±65.98, 172.97±63.51 vs. 179.25±60.44,F=0.349,P=0.706〕. It was shown by NK cell degranulation detection that there was no significant difference in the expression of CD107 and interferon-γ(IFN-γ)secretion〔CD107:0.135±0.050,0.140±0.058,0.128±0.070,F=0.583,P=0.560;IFN-γ(kU/L):14.36±4.74,12.49±4.21, 13.45±5.04,F=1.616,P=0.202〕among healthy control group,SIRS group,and sepsis group. It was shown by antibody dependent cytotoxic effect(ADCC)test that there was no difference in the expression of CD107 among healthy control group,SIRS group,and sepsis group(0.574±0.166,0.643±0.165,0.581±0.157,F=0.808,P=0.448). When compared with healthy controls,the secretion of IFN-γwas increased in SIRS patients(kU/L:40.5±13.2 vs. 28.4±9.6,P=0.001),while reduced in sepsis patients(kU/L:19.8±6.7 vs. 28.4±9.6,P<0.01). Compared with SIRS group,only NK cell surface inhibitory receptors CD158e(KIR 3DL1)expression in sepsis group was significantly increased(0.203±0.057 vs. 0.079±0.021,t=15.762,P<0.001),and there were no significant differences in the other phenotype between the two groups. Compared with SIRS group,the IFN-γproduction of the sepsis group was significantly lowered(kU/L:0.280±0.040 vs. 0.310±0.038,t=3.390,P=0.009),and the level of IL-12 was also significantly decreased(ng/L:0.15±0.03 vs. 0.30±0.08,t=32.832,P<0.001). Conclusion It was showed by NK cell phenotype and function assay that the function of NK cells in patients with sepsis was impaired and led to a poor production of IFN-γ. The IFN-γmediated immune dysfunction may be a main reason for the disorder of NK cell function,which laid the foundation of the clinical immune intervention practice to improve to NK cell function.

Objective:To construct a specific small hairpin RNA (shRNA) expressing vectors against human receptor for advanced glycation end product (RAGE) gene and study its inhibitory effect on the proliferation of androgen-independent prostate cancer cells DU145. Methods:Four RAGE specific oligonucleotides were designed and synthesized. These oligonucleotides were annealed to forill double strand DNA fragments and this fragment was cloned into psi-U6 plasmid. The recombinants were transfected into RAGE-overexpressing sub DU145-2C1 cells. Cellular morphology and transfection efficiency were observed under fluorescence microscope. The inhibitory effect of RAGE shRNA construct on RAGE mRNA and protein expression was examined with semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting, respectively. The cellular proliferation was detected with cell counting kit-8 (CCK-8). Scratch test was used to observe the migration of DU145 cells.Results:RAGE shRNA expression plasmids were successfully constructed and transfected into sub DU145-2C1 cells. It can effectively inhibit the expression of RAGE mRNA (P<0.05). The inhibitory effects of shRNA RAGE-1 (R1) was the most stronger. The RAGE mRNA expression was inhibited by 84% and RAGE protein expression was inhibited by 27%. Compared with negative control, the proliferation potential was significantly decreased in shRNA RAGE-transfected cells. The cell migration capability had no significant changes. Conclusion:RAGE shRNA effectively inhibited the expression of RAGE mRNA and protein and suppressed the proliferation of DU145 cells in vitro.

Objective In order to evaluate the possible effects of myeloid-derived suppressor cells (MDSCs) on graft versus host disease (aGVHD) development and clinical outcomes,this study systematically detected the dynamic changes of MDSCs accumulation in patients during the first 100 days after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Peripheral blood was obtained from 30 patients and 10 healthy volunteers with heparin anticoagulant tubes for 6 mL.For patients,peripheral blood was collected during the first 100 days after allo-HSCT and MDSCs levels were detected by flow cytometry.For measuring the serum concentrations of IL-6,IL-10,IL-1β,TNF-α,Arg-1,HO-1 and iNOS,samples were analyzed using ELISA kits.Results Patients developing aGVHD were infused with significantly less number of MDSCs [(39.94 ± 8.383) 106/kg]than in those not developing aGVHD [(209.0 ± 57.68) 106/kg,P =0.002 6];Patients developing aGVHD Ⅰ-Ⅱ and patients without aGVHD received significantly greater number of MDSCs [(61.96 ± 13.67) 106/kg and (209.0 ± 57.68) 106/kg] than in those developing aGVHD Ⅲ-Ⅳ [(20.37 ±4.304) 106/kg,P =0.013 9].After allo-HSCT,the mean percentage of MDSCs increased markedly in patients developing aGVHD [(7.725 ± 1.460)％] as compared with those not developing aGVHD [(3.423± 1.044)％,P =0.021 3].The high MDSCs group (＞53.712 × 106/kg) showed more favorable clinical outcomes than in the low MDSCs group (≤53.712 × 106/kg).The 2-year overall survival rate as 100％ in high MDSCs group,and 50％ in low MDSCs group (P =0.001 3).The cumulative incidence of 2-year relapse was 6.250％ and 29.252％ in high MDSCs group and low MDSCs group respectively (P =0.112 3).The cumulative incidence of NRM was significantly lower in high MDSCs group (0％) than in low MDSCs group (49.519％,P=0.001 8).MDSCs frequencies significantly increased in patients developing aGVHD after allo-HSCT.After allo-HSCT,the concentrations of IL-6,IL-10,TNF-α,Arg-1,iNOS and HO-1 were significantly elevated in patients developing aGVHD.Conclusion The number of MDSCs when engraftment may be used as a predictor for the development and severity of aGVHD.MDSCs might be considered as a potential new approach to regulate transplant rejection and achieve long-term survival.

Objective To measure volume of breast cancer , calculate tumor volume doubling time (TVDT), and analyze the correlated factors affecting TVDT using three-dimensional ultrasound (3D-US). Methods We applied 3D-US to measure the volume of breast cancer of BI-RADS-US 4A classified by conventional ultrasound. The breast cancer case scanned by 3D-US at least twice (the interval is 3 months at least) without any medical intervention were included in the study. We calculated TVDT according to the formula, and analyzed the affecting factors of TVDT using multiple linear regression. Results Sixty-nine cases were enrolled in the study. The TVDT of breast cancer were from 66 to 521 days , in an average of 185 ± 126 days and the median time of 164 days. We found that: ① there were no statistics significances in TVDT between different breast cancer pattern , smoothing border lines , speculated sign , hyperechoic halo , microcalcification and different rear echo (P > 0.05). ② TVDT of different age groups, lymph node metastasis, pathological grade and NPI score were significantly different (P < 0.05), while menstrual cycle, family history of breast cancer and molecular typing showed no difference (P > 0.05). ③ TVDT of patients with different expression of ER, PR and Ki-67, molecular typing showed statistically difference (P < 0.05), but there was no significant difference between different level of HER2 (P >0.05). ④ multi-factor analysis showed that the NPI score, lymph node metastasis, Ki-67 and molecular typing of breast cancer were relative factors in TVDT (P < 0.05). Conclusions The NPI score , lymph node metastasis , Ki-67 and molecular typing significantly correlate with TVDT of breast cancer. Triple-negative breast cancer in molecular typing has the fastest growth rate.