In order to identify Cimicifugae Rhizoma from its adulterants and to ensure its safe use, the internal transcribed spacer 2 (ITS2) sequence of Cimicifugae Rhizoma and its adulterants were amplified and bidirectionally sequenced by DNA barcoding technology. Sequence assembly and consensus sequence generation were performed by the CodonCode Aligner V3.7.1. The genetic distances were computed by MEGA 5.0. Identification analyses were performed using neighbor-joining (NJ) methods. The length of ITS2 sequence of the three origin plants of Cimicifugae Rhizoma include Cimicifuga heracleifolia, C. foetida, C. dahurica was 217, 219 and 219 bp, respectively. Their intraspecific genetic distance was much lower than the interspecific genetic distance with their closely related species. The NJ tree of ITS2 indicated that the three origin plants of Cimicifugae Rhizoma formed a monophyletic clade, Cimicifugae Rhizoma and its adulterants could be distinguished clearly. The authors proposed that ITS2 sequence was suitable for the authentication of Cimicifugae Rhizoma and its adulterants.
Base Sequence ; China ; Cimicifuga ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics

First with the qualified rate of granules as the evaluation index, significant influencing factors were firstly screened by Plackett-Burman design. Then, with the qualified rate and moisture content as the evaluation indexes, significant factors that affect one-step pelletization technology were further optimized by Box-Behnken design; experimental data were imitated by multiple regression and second-order polynomial equation; and response surface method was used for predictive analysis of optimal technology. The best conditions were as follows: inlet air temperature of 85 degrees C, sample introduction speed of 33 r x min(-1), density of concrete 1. 10. One-step pelletization technology of Biqiu granules by Plackett-Burman design and Box-Behnken response surface methodology was stable and feasible with good predictability, which provided reliable basis for the industrialized production of Biqiu granules.
Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Temperature

With inlet temperature, specific gravity, feeding speed as independent variables, the comprehensive evaluating indexes of content of schisandrin and arctiin as dependent variable, the experimental data were fitted to a second order polynomial equation. Based on establishing the mathematical relationship between the comprehensive evaluating indexes and respective variables, Box-Benhnken central composite test and response surface analysis method was employed to optimize the spray drying technology of Biqiu granules ethanol extract. The optimal drying parameter was as follows: the inlet temperature was 175 degrees C, the specific gravity was 1.10, feeding speed was 32 r x min(-1). Under these conditions, the comprehensive evaluating indexes of spraying dry processes was 92.68, which was close to the model prediction. The spraying dry technology of Biqiu granules ethanol extract optimized by response surface methodology was accurate and feasible, which provided theoretical experiment basis for the industrialization production.
Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ethanol

The intestinal absorption of berberine (Ber) and its structural modified compound 8-hydroxy dihydroberberine (Hdber) was compared, and their effects on the intestinal absorption of sugar by perfusion experiment were investigated in order to reveal the mechanism of low dose and high activity of Hdber in the treatment of hyperglycemia. The absorption of Hdber and Ber in rat small intestine was measured by in situ perfusion. High performance liquid chromatography (HPLC) was used to determine the concentrations of Hdber and Ber. In situ perfusion method was also used to study the effects of Hdber and Ber on sugar intestinal absorption. Glucose oxidase method and UV spectrophotometry were applied to examine the concentrations of glucose and sucrose in the perfusion fluid. The results showed that the absorption rate of Ber in the small intestine was lower than 10%, but that of Hdber was larger than 70%. Both Hdber and Ber inhibited the absorption of glucose and sucrose at the doses of 10 and 20 μg/mL. However, Hdber presented stronger activity than Ber (P<0.01). It is suggested that Hdber is absorbed easily in rat small intestine and that its inhibitory effect on the absorption of sugar is better than Ber.

AIM: To identify the novel nuclear export signal by analyzing the DNA sequences and detecting the cell localization of different adenovirus ElA - associated protein BS69 isoforms. METHODS:BS69 DNA sequences in Emsebl database were blasted and the sequence of amino acids was aligned with the typical nuclear export signal. Different BS69 isoform fragments were cloned into pcDNA3.1 vector and transfected into Cos7 cells. The BS69 localization was observed by immunostaining and the function was verified by Western blotting. RESULTS: A novel nuclear export signal was found in BS69 isoform 2 but not in isoform 1.The isoform 2 was localized in cytoplasm and isoform 1 in nucleus, which was also consistent with the DNA sequence. The isoform 2 was involved in LMP1 - activated JNK phosphorylation whereas the isoform 1 was not. CONCLUSION: Different BS69 isoforms have different cellular localization. BS69 isoform 2 is localized in cytoplasm, interacting with Epstein - Barr virus latent membrane protein 1 and may be involved in nasopharyngeal carcinoma development.However, the isoform 1 is localized in nucleus and plays important roles in transcription.

Objective:This study aims to analyze the differences in the alternative splicing pattern of ADAR2 among glioma cell lines U87, U251, A172, and normal human astrocyte HA1800. Methods:A-to-I editing level at the Q/R-Site of GluR-2 was analyzed by RT-PCR and sequencing. Real-time PCR was performed to detect the expression level of each alternatively splicing variant using a specific primer that was confirmed to amplify only the targeted template and not other alternatively spliced variant fragments. Results:We verified that the Q/R-Site of GluR-2 is under-edited in glioma cell lines. Real-time PCR revealed that the ADAR2 pre-mRNA splic-ing pattern has no significant difference at exons 1a and 2 between glioma cell lines and normal human astrocyte. We also detected that the amount of alternative splicing variants, including exon 5a, was higher than that of alternative splicing variants not including exon 5a in human glioma cell lines. However, the expression of alternative splicing variants, including exon 5a, was lower than that of alterna-tive splicing variants not including exon 5a in human astrocyte. Conclusion:Evident differences in splicing were observed at the site of exon 5a between glioma cell lines and normal human astrocytes. The difference in the alternatively splicing pattern at exon 5a may be attributed to the decreased activity of ADAR2.

Objective To explore the value of abnormal imaging findings on susceptibility weighted imaging (SWI) in delayed graft function (DGF). Methods The conventional MRI and SWI images of 26 cases with DGF and 20 cases with normal renal function of transplanted kidneys were retrospectively analyzed. Patients with cysts and angiomyolipomas were excluded. Normal structures of transplanted kidney were identified. If lesions of abnormal signal intensity were found in the transplanted kidney, the location, border and signal intensity compared to renal cortex would be analyzed. The differences in signal intensity between the abnormal signal lesions and normal renal cortex in the same SWI layer of DGF were compared by using independent-sample t test. The differences in positive detection rate of discovering the abnormal signal lesions in DGF between conventional MRI and SWI were compared by using McNemar test. Results Of the 26 cases with DGF, one case was found to exhibit abnormally low signal lesions with fuzzy boundary located at junctional zone between cortex and medulla on both conventional MRI and SWI images. Ten cases were found to exhibit abnormally low signal lesions with fuzzy boundary located at junctional zone between cortex and medulla on SWI images only. Fifteen cases exhibited no abnormal signal lesions on both conventional MRI and SWI images. Twenty cases with normal renal function of transplanted kidney, no abnormal signal lesions were found on both conventional MRI and SWI images. The differences in signal intensity between the abnormally low signal lesions (130±20) and normal renal cortex (177±25) in the same SWI layer of 11 cases with DGF were statistically significant (t=-4.582,P<0.01). The differences in positive detection rate of discovering the abnormally low signal lesions in DGF between conventional MRI [3.8%(1/26)] and SWI [42.3% (11/26)] were statistically significant (χ2=8.100,P=0.002). Conclusions Abnormally low signal lesions with fuzzy boundary located at junctional zone between cortex and medulla on SWI images suggest the presence of DGF. Compared with conventional MRI, SWI appears to be superior in detecting the abnormally low signal lesions.

Objective To investigate the clinical and MRI characteristics of reversible splenial lesion syndrome (RESLES).Methods The clinical data,MRI features and follow-up results of 18 patients with RESLES were analyzed retrospectively.Results The average age of the patients was younger,and 5 cases were below 1 5 years old.Clinical history and MRI features were characterized.About 67%(12/18)patients had fever,headache,encephalitis,history,without long history of alcohol abuse.MRI revealed that the lesions of splenial of corpus callosum were solitary nodule with clear boundary.The lesions were slightly hyperintensity on T2 WI and obviouslyhyperintensity on FLAIR and DWI,while the they show hypointensity on ADC with the ADC value <1×10-3 mm2/s.No edema or significant enhancement were observed.The lesions may completely disappear or relief on DWI after short treatment.Conclusion RESLES is a rare entity with a broad clinicoradiological spectrum.The MRI features are characteristic,and it will provide helpful information for diagnosis, differential diagnosis and clinical treatment.

OBJECTIVETo explore the effect of qidong huoxue decoction (QHD) on inflammatory factors and Toll-like receptor (TLR4) mRNA expressions in acute lung injury (ALI) rats.

METHODSTotally 50 healthy male SD rats were randomly divided into the blank control group, the lipopolysaccharide (LPS) model group, low, middle, high dose QHD groups according to body weight, 10 rats in each group. Rats in low, middle, high dose QHD groups were intragastrically administered with QHD at 4, 8, and 16 mL/kg 24, 12 h before modeling and 12 h after modeling, respectively. Normal saline was intragastrically administered to rats in the blank control group and the LPS model group. An ALI rat model was established using intratracheal instillation of LPS. Rats were killed after 24-h modeling. Then the bronchoalveolar lavage fluid was prepared. Contents of TNF-α, IL-1β, and L-10 were detected using ELISA. TLR4 mRNA expressions were determined byreal time PCR.

RESULTSCompared with the blank control group, contents of TNF-α, IL-1β , and IL-10 increased (P <0. 01), TLR4 mRNA expressions also increased in the LPS model group (all P <0. 01). Compared with the LPS model group, contents of TNF-α and IL-1β decreased (P <0. 05, P <0. 01), IL-10 levels increased (P <0. 01) , TLR4 mRNA expressions were also reduced (P <0. 01), in high and middle dose QHD groups. Compared with the high dose QHD group, con- tents of TNF-α and IL-1β increased in middle and low dose QHD groups (P <0. 05); IL-10 levels decreased (P <0. 05) in the low dose QHD group(P <0. 05), TLR4 mRNA expressions also increased in the low dose QHD group (P <0. 05). Compared with the middle dose QHD group, IL-10 levels was reduced, but TLR4 mRNA expressions increased in the low dose QHD group (P <0. 05).

CONCLUSIONSQHD had the protective effect on LPS induced ALI rats. Its mechanism might be associated with inhibiting TLR4 mRNA expressions, leading to decreased pro-inflammatory cytokines such as TNF-α and IL-β, elevated anti-inflammatory cytokine IL-10, and thereby, correcting unbalanced inflammation.

Acute Lung Injury ; genetics ; metabolism ; Animals ; Anti-Inflammatory Agents ; Bronchoalveolar Lavage Fluid ; Drugs, Chinese Herbal ; pharmacology ; Inflammation ; Interleukin-10 ; metabolism ; Interleukin-1beta ; metabolism ; Lipopolysaccharides ; Male ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

The effect of amygdalin joint hydroxysafflor yellow A (HSYA) on the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1beta and the possible mechanism were studied and explored. Chondrocytes were obtained from endplate of one-month SD rat intervertebral discs and cultured primary endplate chondrocytes. After identification, they were divided into normal group, induced group, amygdalin group, HSYA group and combined group. CCK-8 kit was adopted to detect the proliferation of the endplate chondrocytes. FCM was measured to detect the apoptosis. Real-time PCR method was adopted to observe the mRNA expression of Aggrecan, Col 2 alpha1, Col 10 alpha1, MMP-13 and the inflammatory cytokines IL-1beta. The protein expression of Col II, Col X was tested through immunofluorescence. Compared with the normal group, the proliferation of the endplate chondrocytes decreased while the apoptosis increased (P < 0.05). With down regulation of the mRNA expressions of Aggrecan, Col 2 alpha1 and up regulation of the mRNA expressions of Col 10 alpha1, MMP-13, IL-1beta (P < 0.05), the protein expression of Col II decreased while the protein expression of Col X increased. Compared with the induced group, amygdalin group, HSYA group, the combined group could inhibit the apoptosis and promote the proliferation (P < 0.05). They could increase the mRNA expressions of Aggrecan and Col 2 alpha1 while decrease the mRNA expressions of Col 10 alpha1, MMP-13 and IL-1beta (P < 0.05). They could also enhance the protein expression of Col II while reduce the protein expression of Col X. The effect of the combined group was significantly better than that of amygdalin and HSYA. Amygdalin joint HSYA could inhibit the degeneration of the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1beta and better than the single use of amygdalin or HSYA.
Amygdalin ; pharmacology ; Animals ; Apoptosis ; Cells, Cultured ; Chalcone ; analogs & derivatives ; pharmacology ; Chondrocytes ; drug effects ; Collagen ; metabolism ; Drug Synergism ; Interleukin-1beta ; Intervertebral Disc ; cytology ; Quinones ; pharmacology ; Rats