Humans ; MicroRNAs ; Non-alcoholic Fatty Liver Disease

OBJECTIVE:To establish the quality standard for Qindan granules. METHODS: The constituents including Radix Scutellariae, Radix Astragali, Cortex Moutan and Fructus Gardeniae in Qindan granules were identified respectively by TLC. RESULTS: The test samples and their respective reference substances were alike in that the same color chromatographic spots were noted at the corresponding places. The color spectra of the samples were clear yet without interference of surrounding impurity. CONCLUSION: The established method has high specificity and can be used for the quality control of Qindan granule.

Objective The regulation of ornithine decarboxylase (ODC) gene expression and enzyme activity by corticosterone, the main glucocorticoid in rat, during rat liver regeneration induced by partial hepatectomy (PH) was evaluated.Methods Bilateral adrenaleetomies (ADX) and sham-ADX were performed on ether-anesthetized rats 3 days before PH.Corticosterone in sesame oil was injected subcutaneously to adrenalectomied rats. ODC mRNA, ODC protein and enzyme activity were detected by RT-PCR, Western blotting and high performance liquid chromatography (HPLC), respectively. Results The ODC mRNA levels, protein accumulation and enzyme activity were lower in the intact liver compared to the regenerating liver.After PH, mRNA levels were remarkably enhanced in all groups (n=6 in each group) and peaked at 5 hours post-PH. Till 7 hours, the contents in all groups from high to low were ADX group,control group (Sham-ADX group), ADX treated with 10mg/kg and 40mg/kg body weight corticosterone group, respectively. ODC protein accumulation in ADX rats was higher than that in control rats (n=13, the same below), but it decreasod in corticosterone-treated (10mg/kg) rats until 24 hours post-PH, with a strong decline seen in 40mg/kg corticosterone-treated rats. ODC activity was rapidly promoted, and the highest levels were observed at 6 hours after PH in all groups (n=6 in each group). After corticosterone treatment, the activities declined significantly at 6 hours post-PH, with the lowest value found in the 40mg/kg group. Conclusion Corticosterone treatment results in dose-dependent decreases in ODC mRNA and enzyme protein both in the intact liver and the regenerating liver. The change in ODC activity is partially related to alterations of ODC mRNA and protein accumulation.

Objective To investigate the effects of fentanyl and remifentanil on the viability of human adenocarcinoma cell line A549.Methods Human adenocarcinoma A549 cells cultured in logarithmic growth phase were seeded in 75 ml culture bottles or 96-well plates.After being cultured for 24 h,the cells were randomly divided into 9 groups (n =30 each):4 fentanyl groups (groups F1-4 ),4 remifentanil groups (groups RF1-4 ) and control group (group C).Groups F1-4 were exposed to fentanyl with the final concentrations of 0.5,5.0,50.0 and 500.0 ng/ml respectively.Groups RF1-4 were exposed to remifentanil with the final concentrations of 0.5,5.0,50.0 and 500.0 ng/ml respectively.The viability of the cells was determined by methyl thiazolyl tetrazolium assay after being incubated for 24,48 and 72 h.The cell cycle progression and apoptosis were determined by flow cytometry after being incubated for 24 h.Results Compared with group C,the viability of A549 cells were gradually decreased at 72 h of incubation,the proportion of the cells in S phase was gradually decreased at 24 h of incubation,and the proportion of the cells in G2/M phase and apoptotic rate were gradually increased in groups F2-4 and in groups RF2-4 ( P ＜ 0.05).Conclusion Fentanyl and remifentanil with the final concentration ≥5 ng/ml can inhibit the viability of human adenocarcinoma cell line A549 in a dose-independent manner by inducing cell apoptosis and cell cycle arrest in G2/M phase.

Aim To synthesize and identify artificial antigen of podophyllotoxin for the production of podophyllotoxin polyclonal antibody. Methods The hapten was synthesized by two different chemical approaches and characterized by TLC, IR, NMR, and MS. Mixed anhydride reaction (MAR) and active ester method (AEM) were used to couple the podophyllotoxin to carrier proteins (BSA and OVA). Characterization of artificial antigens was done by using spectroscopy and electrophoresis. The anti-podophyllotoxin polyclonal antibodies were obtained through immunizing rabbits. Results The results from IR, NMR and MS showed that 4-O-succinoyl podophyllotoxin (hapten) was successfully synthesized. The coupling molar ratios of the hapten and carrier proteins were 88.6 for Hapten-BSA1, 40.3 for Hapten-BSA2, 17.8 for Hapten-OVA1, and 54.2 for Hapten-OVA2. Hapten conjugates coupled with BSA yielded two sets of the specific and affinitive polyclonal antibodies. One set of antibodies showed an IC50 value of 2.21 μg·mL -1 with a detection limit of 0.12 μg·mL -1. Conclusion Antigenic conjugates were artificially synthesized, and based on these artificial antigens, polyclonal antibodies against podophyllotoxin were raised from rabbits immunized with two different immunogens and characterized with an indirect ELISA format.

Objective To establish the multiple quantitative fluorescent polymerase chain reaction (QF-PCR)assay and evaluate its clinical application in prenatal diagnosis.Methods Totally 170 samples Were collected between May 2008 and July 2009 in prenatal center of Peking Union Medical College Hospital:123 of them were amniotic fluid,9 were chofionic villous samples,20 were fetal blood and 18 were villi from aborted fetuses.All samples were from women of Han nationality,with mean age of (34.1±4.6) years old,and with mean gestational age of(19.6±1.0)weeks.Cytogenetic cultures and karyotyping were made to every sample.Genomic DNA wag extracted from the samples.The sequences of twenty short tandem repeat (STR) markers were designed according to the GenBank and references,including 6 STR markers in chromosome 21.4 in chromosome 18.4 in chromosome 13,4 in chromosome X,1 in chromosome Y and 1 universal marker in both X and Y chromosome.Each sample was amplified by two sets of multiple QF-PCR,which included 4 STR markers in each of 21,18,13 and sex chromosomes. If the result was uninformative,the third set of anotherd 4 STR markers was added. Results ( 1 ) Karyotyping. Cytogenetic analysis were made for all the 170 samples, 151 (89%) of which were normal, and 19 (11% ) were abnormal (2)QF-PCR assay. 167(98% ) samples were detected by QF-PCR. The results were obtained within 2 -3 days after sampling. 134 samples were proved normal by QF-PCR, which was consistent with karyotyping. Among the 19 abnormal karyotype samples, 18 were detected as abnormal( eight were 21-trisomy, three were 18-trisomy)by QF-PCR. Among the 167 samples, 150(90% ) were detected using the first and second set of STR mixtures, and 3(2% ) were detected when the third set of STR was added. The remain 14(8% ) were uninformative. (3) The diagnostic efficiency of QF-PCR. The sensitivity of QF-PCR in prenatal diagnosis of common aneuploidities was 95%, the specificity, the false positive rate, the false negative rate, the positive predictive value and negative predictive value were 100% ,0,5%, 100% and 99% , respectively. (4)Autusome and sex chromosome detection by QF-PCR. Among all the STR markers,D21S1270 and D21S1411 had the highest heterozygosifies in chromosome 21, and DXS8377 had the highest in sex chromosome. The amplifications were stable. Conclusion Multiple QF-PCR assay is a valid alternative in rapid prenatal diagnosis of common chromosome aneuploidies. With high accuracy, it can be used for numerous sample test in large-scale laboratories.

Objective To evaluate the effect of chemotherapy on sedation with propofol in breast cancer patients.Methods One hundred female patients,of ASA physical status Ⅰ or Ⅱ,aged 20-60 yr,scheduled for elective modified radical mastectomy,were divided into 2 groups (n =50 each) according to whether receiving neoadjuvant chemotherapy before operation:non-chemotherapy group (group Ⅰ) and neoadjuvant chemotherapy group (group Ⅱ).The breast cancer patients received operation directly in group Ⅰ.The breast cancer patients received neoadjuvant chemotherapy in group Ⅱ.Epirubicin 75-100 mg/m2 was injected intravenously on 1st and 2nd days,docetaxel 75 mg/m2 was injected intravenously on 3rd day,and 3 weeks were considered as 1 course of treatment.The patients received operation at 3 weeks after the end of 4 courses of treatment in group 1.Anesthesia was induced with propofol given by target-controlled infusion and the target plasma concentration of propofol was 3.5 μg/ml.The time for loss of consciousness and consumption of propofol at loss of consciousness were recorded.Results Compared with group Ⅰ,the time for loss of consciousness was significantly shortened,and the consumption of propofol at loss of consciousness and BIS value were decreased in group Ⅱ.Conclusion Chemotherapy can enhance propofol-induced sedation and promote the onset of propofol in breast cancer patients.

Objective To evaluate the effect of sleep dysfunction on sedation induced by propofol in the patients undergoing radical mastectomy.Methods One hundred breast cancer patients,aged 25-60 yr,with body mass index of 19-23 kg/m2,of ASA physical status Ⅰ or Ⅱ,scheduled for elective modified radical mastectomy,were randomly divided into 2 groups according to sleep quality.The patients with global Pittsburgh Sleep Quality Index (PSQI) score ≤7 served as regular sleep quality group (Ⅰ group,n =59).The patients with global PSQI score ＞ 7 served as sleep dysfunction group (group Ⅱ,n =41).Anesthesia was induced with propofol given by target-controlled infusion (target plasma concentration of 3.5 μg/ml),and then with remifentanil 4 μg/kg and rocuronium 0.6 mg/kg after loss of consciousness.The consumption of propofol at loss of consciousness was recorded.Results Compared with group Ⅰ,the consumption of propofol at loss of consciousness was significantly decreased in group Ⅱ.Conclusion Sleep dysfunction can enhance propofol-induced sedation in the patients undergoing radical mastectomy.

BACKGROUND: Osteogenic ability of bone morphogenetic protein-2 has been wel documented in many experiments, but a series of factors are involved in osteogenesis induction that is a complex network adjustment process. OBJECTIVE: To quantitatively determine the level of insulin-like growth factor I during the lumbar spinal fusion of rabbits induced by recombinant human bone morphogenetic protein-2. METHODS: Sixty adult male New Zealand white rabbits were randomly divided into three groups: bone autograft, bone al ograft or composite bone (bone al ograft with 75 μg recombinant human bone morphogenetic protein-2) was implanted into the L5-6 intertransverse process of rabbits, respectively. At days 7, 14, 21, 28, 35 after implantation, formed cal us was taken to detect the expression of insulin-like growth factor I using real-time fluorescence quantitative PCR. RESULTS AND CONCLUSION: In the three groups, the expression of insulin-like growth factor I gradual y increased with implantation time, peaked at 28 days and then decreased. At 7 days after implantation, the expression of insulin-like growth factor I was higher in the autograft group than the composite and al ograft groups (P < 0.05); at 14 days, the expression of insulin-like growth factor I was higher in the autograft and composite groups than the al ograft group (P < 0.05); at 21, 28 and 35 days, the expression of insulin-like growth factor I was higher in the composite group than the autograft and al ograft groups (P < 0.05). These findings indicate that recombinant human bone morphogenetic protein-2 can improve the expression of insulin-like growth factor I effectively during the lumbar spinal fusion.

Air Pollutants, Occupational ; chemistry ; Indium ; analysis ; Mass Spectrometry ; methods ; Occupational Exposure ; Spectrum Analysis ; methods ; Workplace