Objective To study the expression profile of microRNA (miRNA) and the roles in pathogenesis of acute liver failure in mouse model. Methods Eighty-five BALB/c mice were divided into four groups: 40 in model group of acute liver failure were intraperitoneally injected with Dgalactosamine (D-GalN) and lipopolysaccharides (LPS); 20 in D-GalN group were injected with DGalN only; 20 in LPS group were injected with LPS only; 5 in control group were injected with saline.Liver histology of mouse was observed at hour 0, 5, 7 of injection, and sera and liver tissues were collected at hour 0, 1, 3, 5, 7, 9 of injection. Meanwhile, levels of inflammatory factors [tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)] in serum and liver tissue were detected by realtime polymerase chain reaction (PCR). Lock nucleic acid (LNA)-based miRNA microarray technology was used to detect the expression profile of hepatic miRNA, and the expression of miRNA was verified by real time quantification-polymerase chain reaction (RT-PCR). Mouse macrophage Raw264.7 cells were induced by LPS in vitro and the expressions of miRNA at different time points were detected.The comparison of means among groups was analyzed using one way ANOVA and the correlation were analyzed by Pearson and Spearman correlation. Results Microarray analysis found that the expression profile of miRNA during the acute liver failure changed dramatically. There were 97 miRNA in model group changed significantly compared with control group (P＜0.01), including 21 up-regulated and 27down-regulated at hour 5 and 7 of injection. Furthermore, the expressions of miR 146a and miR-155were verified by RT-PCR and found they both increased progressively over time after injection.Correlation analysis showed that miR-155 was well correlated with both TNF-α and IL-6 expressions.It was further found that miR-146a and miR-155 were both up-regulated in activated Raw264.7 cells in vitro. Conclusions The expression profile of miRNA changes during acute liver failure in mouse model. Inflammation associated-miR-146a and miR-155 are both up-regulated significantly, which indicatcs that they may play an important regulatory role in pathogenesis of acute liver failurc.

Traditional herbal medicines, Panax ginseng, Panax quinquefolium and Panax notoginseng, attract our attention for their extensive and powerful pharmacological activities. Ginsenosides are the main active constituents of these medicinal herbs. The related glycosyltransferases involved in ginsenoside biosynthesis are the key enzymes which catalyze the last important step. Modification of ginsenoside aglycones by glycosyltransferases produces the complexity and diversity of ginsenosides, which have more extensive pharmacological activity. At present, ginsenoside aglycones and compound K have been obtained by synthetic biology. As the last step of ginsenoside biosynthesis, glycosylation of ginsenoside aglycones has been studied intensively in recent years. This review summarizes the basic strategies and research advances in studies on glycosyltransferases involved in ginsenoside biosynthesis, which is expected to lay the theoretical foundation for the in-depth research of biosynthetic pathway of ginsenosides and their production by synthetic biology.
Biosynthetic Pathways ; Ginsenosides ; biosynthesis ; Glycosyltransferases ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Synthetic Biology

Objective To investigate the differential value of contrast-enhanced ultrasonography (CEUS) and time-intensity curves(TIC) in diagnosing testicular and epididymal mass lesions.Methods CEUS via intravenous bolus injection of SonoVue and TIC were performed for quantitative analysis of testicular and epididymal mass lesions.Forty-one patients with 42 testicular and epididymal mass lesions and 26 normal testicles were examined with CUES,the perfusion progress were recorded dynamically,which findings were compared with surgery.Results Twenty-three (54.76％) malignant masses displayed enhanced pattern as evenly enhanced,fast-in and fast-out (8.7％),evenly enhanced,fast-in and slow-out (65.2 ％ ),unevenly enhanced,fast-in and slow-out (26.1％ ).Ninteen (45.24 ％ ) benign masses revealed enhanced pattern as unevenly enhanced,fast-in and slow-out ( 10.5 ％),evenly enhanced,slow-in and slowout ( 10.5 ％ ),unevenly enhanced,slow-in and slow-out ( 36.9 ％ ),without enhancement ( 42.1％ ).There was statistical difference of peak intensity,time to peak and areas among malignant group,benign group and normal group ( P ＜ 0.05).Conclusions CEUS combined with TIC could provide differential diagnostic value for testicular and epididymal mass lesions.

Objective To explore the relationship between gene mutation and different syndromes of TCM in the early stage of diabetic nephropathy(DN).Methods Sixty-three patients with diabetic nephropathy in the early stage were observed.The methylene tetrahydrofolate reductase(MTHFR)C676T polymorphism was determined by polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP)assay.The levels of homocysteine(Hcy),acidum folicum,fasting glucose,postprandial glucose,glycosylated hemoglobin(HbA1C),urinary albumin excretion rate(UAER),and blood lipid were measured respectively,and the TCM syndromes were recorded.Results Of the 63 patients,19 were genotype CC,17 were genotype TT,and 27 were genotype CT.The genotypic frequency of TT was 27.00%,that of CT was 42.85%,and that of CC was 30.15%.The T allele frequency was 48.41% and C allele frequency 51.59%.The MTHFR C676T mutation was related with plasma Hcy level(P

Objective To evaluate the relationship of clinical symptom to plasmic levels of D-dimer, activated factorⅦ (FⅦa) and tissue factor pathway inhibitor (TFPI)/X a in patients with urticaria. Methods A total of 27 patients with chronic urticaria (CU), 27 patients with acute urticaria (AU) and 26 normal human controls were included in this study. Symptom score was determined and disease course was surveyed in these patients. ELISA was used to detect the plasma levels of D-dimer, FⅦa and (TFPI)/Xa in patients and controls. The relation of clinical symptom and disease course to plasma levels of these parameters was assessed. Results In patients with AU and normal controls, the plasma level of D-dimer was 450.57± 242.13 ng/mL and 266.81±40.68 ng/mL, respectively, the level of FⅦa, 2.23± 0.74 ng/mL and 5.23±1.35 ng/mL, respectively, and the level of TFPI/Xa 0.87±0.13 nmol/L and 0.88 ~ 0.12 nmol/L, respectively. There was a significant difference in the level of both D-dimer and FⅦa (both P < 0.01 ), whereas no differ-ence was observed in that of TFPI/X a (P > 0.05) between patients with AU and normal controls. In addi-tion, increased level of D-dimer and decreased level of FⅦa were noticed in patients with CU compared with those in normal controls (593.80±294.04 ng/mL vs 266.81±40.68 ng/mL, 3.98±0.35 ng/mL vs 5.23± 1.35 ng/mL, both P < 0.01 ), but there was no significant difference in the plasma level of TFPI/Xa (0.87± 0.16 nmol/L vs 0.88±0.12 nmol/L, P > 0.05). Significant difference was observed in the plasma level of D-dimer and FⅦa between patients with AU and CU (450.57±242.13 ng/mL vs 593.80 ±294.04 ng/mL, P < 0.05; 2.23± 0.74 ng/mL vs 3.98± 0.35 ng/mL, P<0.01 ). The plasma level of D-dimer positively corre-lated to the symptom score of patients with CU and those with AU (r= 0.68, P< 0.01; r= 0.82, P< 0.01),but was independent of discase course (P> 0.05). Neither the level of FⅦa nor that of TFPI/Xa correlated to symptom score or disease course of patients (all P > 0.05). Conclusions There is an overactivation of coagulation cascade, consumption of blood coagulation factors and secondary fibrinolysis in patients with urticaria, suggesting that plasma D-dimer and FⅦa may be associated with the clinical symptoms of urticaria.

To observe the clinical results of innercapsular phacoemulsification with primary intraocular lens ( lOL ) implantation in the treatment of lens dislocation.METHODS: A total of 23 cases ( 23 eyes ) of lens dislocation ( lla and llb ) were underwent innercapsular phacoemulsification combined with primary lOL implantation. lOL implantation were underwent during operation, the complications of intraoperative and postoperative, postoperative vision, intraocular pressure ( lOP ) , corneal endothelial cell, lOL location were analyzed.RESULTS: The operations were successfully completed for all patients in accordance with the pre - surgery program; lens nucleus or its fragments did not crash into the vitreous cavity; 20 cases of corneal edema and 17 cases of lOP presented at the first day after surgery, the deviation or displacement of lOL and serious complications such as retinal detachment were not appeared. At the first week postoperation, the average lOP was 15. 81 ± 2. 10mmHg, with statistically significant differences when compared with the preoperative ( P<0. 01 ) , the visual acuity in all eases increased, with statistically significant differences when compared with the preoperative ( P < 0. 01 ). Corneal endothelial cell density and percentage of hexagonal cells decreased, the variation coefficient increased in first week of postoperative, with no statistically significant differences when compared with the preoperative (P>0. 05) CONCLUSlON: lnnercapsular phacoemulsification combined with primary lOL implantation in the treatment of the whole lens dislocation (‖a and ‖b ) can restore function in patients with diplopia and control lOP effectively, reduce corneal endothelial cell damage, which is an effective method to treat the whole traumatic lens dislocation.

Objective To investigate the current situation of the prevention and contml of iodine deficiency disorders(IDD)in Hubei Province,SO as to provide a policy.making basis for controlling work of IDD. Methods Using the method of proportional population sampling(PPS),30 eounties were seiected in Hubei Province.In each selected county,1 primary school was chosen.In every primary school,40 pupil8 aged 8～10 years were selected to examine thyroid size,intelligent quotient(IQ),and salt iodine contents at their home.In the selected pupils,2 boys and 2 girls were chosen to determine their urinary iodine contents in each age grouD.Twenty pupils in the above school and 5 housewives ileal"to this school were tested in health education questionnaire. Results The median of salt iodine was 30.1 mg/kg,and the rate of comsuming qualified iodized salt was 96.2% (1154/1200).The rates ofchild goiter were 6.5%(78/1200)by palpation and 4.1%(49/1200)by B ultrasound.The median of urinary iodine Was 358.4μg/L and mean of the IQ was 105.3±14.4.The rate of qualified scores of both students and housewives Was 25.5%(153/600)、90.7%(136/150).Conclusions The current Bituation of iedine nutrition is good.The goal of eliminating IDD has been achieved in Hubei Province.

OBJECTIVETo investigate the role of transcript factor SCL/TAL-1 gene in the erythroid differentiation through the knockdown of SCL/TAL-1 mRNA by RNA interference.

METHODSThe plasmid of pTRIP-dU3-RNAiTALh-EF1a-GFP with SCL/TAL1 shRNA was transfected into EPO-induced K562 cell line with erythroid differentiation via lentiviral vector system and the expression of SCL/TAL-1 mRNA decreased. The plasmid pTRIP-dU3- RNAiluc-EF1-GFP expressing EGFP gene was as control. The mRNA levels of SCL/TAL-1 and erythroid related RhD, GPA, CD47 in the cell lines were detected by RT-PCR, and erythroid antigen CD71, CD235a were examined by flow cytometry.

RESULTS(1) After 48 h of transfect, more than 95% of K562 cells were GFP positive, indicating the infection rate of the plasmids in the K562 cells more than 95%. (2) The results of RT-PCR showed SCL/TAL-1 mRNA expression in the K562 cell line of knockdown of SCL/TAL-1 was significantly lower than that in the control (P < 0.05). The mRNA levels of CD47 and RhD were also significantly lower, however, GPA decreased slightly in comparison with the control. (3) The expressions of CD71 and CD235a markedly reduced in the K562 cell line of knockdown of SCL/TAL-1 with positive rates as 10.4% and 76.5%, while the positive rates in the control as 94.3% and 83.6%.

CONCLUSIONOur findings suggested that transcription factor SCL/TAL-1 might play an positive role in erythroid differentiation.

Cell Differentiation ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; K562 Cells ; RNA Interference

OBJECTIVETo investigate the anti-leukemia effect of oridonin on Ph(+) acute lymphoblastic leukemia (ALL) cell line SUP-B15.

METHODSHuman Ph(+) ALL cell line was cultured in vitro. The 50% inhibition concentration (IC(50)) of oridonin against SUP-B15 cell line was examined using modified MTT assay. The cellular morphologic changes were observed using a light microscope. The percent of apoptosis of SUP-B15 cell line after drug treatment was evaluated by flow cytometric analysis. The active levels of ABL kinase and its downstream Akt/mTOR, Raf/MEK/ERK, STAT5 signaling pathways and the expression levels of Bcl-2 and BAX were examined by Western blot.

RESULTSOridonin inhibited the growth of SUP-B15 cell line in both time- and dose-dependent manner with the IC(50) of oridonin as (7.08 ± 1.21) µmol/L after 72 h treatment. The cellular membrane of SUP-B15 cell line treated with oridonin became unsharp, some of them disintegrated. Oridonin induced apoptosis in SUP-B15 cell line with the apoptosis rates following 0, 5, 10 µmol/L oridonin treatment for 24 h were (6.67 ± 0.83)%, (18.30 ± 1.79)% and (37.63 ± 7.12)%, respectively. Oridonin inhibited activation of ABL kinase and its downstream Akt/mTOR, Raf/MEK/ERK and STAT5 signaling pathways, which were constitutively activated in SUP-B15 cell line, down-regulated the level of anti- apoptotic protein Bcl-2 and up-regulated the expression of pro-apoptotic protein Bax.

CONCLUSIONOridonin exerted anti-leukemia effect in Ph(+)ALL cell line SUP-B15 by inhibiting the activation of ABL kinase and its downstream Akt/mTOR, Raf/MEK/ERK and STAT5 signaling pathways, down-regulating the expression of Bcl-2 and up-regulating the expression of BAX.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diterpenes, Kaurane ; pharmacology ; Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Signal Transduction

Objective To investigate the expressions of microRNAs (miRNA) and cytokines in the peripheral blood mononuclear cells (PBMCs) of patients with hepatitis B virus (HBV) infection and analyze their relationship with inflammation. Methods PBMCs were collected from acute hapatitis B (AHB) patients of 3 groups, including acute episode, virus clearance, recover period, and chronic hepatitis B (CHB) of mild, medium, severe type, and HBV-related liver cirrhosis. Each group included 20 patients, and 17 healthy donors were as control. Reverse transcription-polymerase chain reaction was used to measure miRNA146, miRNA155, miRNA181, interferon (IFN)-α, IFN-β,interferon induced gene 54 (ISG54) and interferon regulate factor 5 (IRF5). Comparisons among groups were done by one factor analysis of variance. Results The expression of miRNA155 was high in acute episode of AHB (2. 386± 1. 835), and higher than healthy control (1. 498± 1. 276) (F=1. 137,P-=0. 045), while reduced in acute episode, virus clearance (1. 633±2. 291), and recover period (0. 642±0. 836) (F=2. 122,P=0. 022). The expressions of IFN-α and IFN-β in AHB reduced in acute episode, virus clearance and recover period ( F = 1. 880, P = 0. 038 ; F= 1. 835, P = 0. 048).The expression of miRNA155 in AHB is closely correlated with IFN-α and IFN-β (r = 0. 483, P=0. 004; r= 0. 660, P= 0. 0002, respectively). In addition, in HBV infectious patients, the expression of miRNA155 was correlated with alanine transarninase (ALT), serum bilirubin (TBil) (r=0. 342,P=0. 006; r=0. 322, P= 0. 011, respectively), but not with HBV DNA load. Compared with healthy control (1. 307+ 0. 935), miRNA181 was higher in patients with HBV infection (F= 2. 072, P=0. 045) except AHB in recover period (1. 873±0. 998). There was no statistical difference in the miRNA146 expression between various groups. Conclusions The exprossion of miRNAs might be involved in the host anti-HBV immune response during HBV infection, and closely correlated with expression of IFN-related immune factors.