Differential Expression and Signal Transduction Analysis of MicroRNA Related to Kidney Yang Deficiency in Kidney Tissues of Rats / 中国实验方剂学杂志

Objective: Microarray chip was used to detect the differentially expressed microRNA (miRNA) in kidney tissues of rats with kidney-Yang deficiency induced by adenine,and its significance was analyzed by bioinformatics method. Method: Rats with kidney-Yang deficiency were induced by intragastric administration of 150 mg·kg-1 adenine in model group, while rats in normal group were given the same amount of saline.Kidney tissues were taken for hematoxylin-eosin (HE) staining pathological sections after anesthesia and blood urea nitrogen (BUN), creatinine (SCr) in blood and 24-hour urinary protein (24U-TP) in urine were measured. μParaflo microfluidic chip technology was used to investigate differential expression miRNA in kidney tissues, and microarray results were verified by Real-time PCR. Bioinformatics database was used to analyze the target genes and functions of differential expression miRNAs. Result: Gene chip results showed that there were 50 differentially expressed microRNAs after modeling. Compared with control group, only 9 miRNAs were highly expressed in kidney tissues with significant difference were detected in model group. Compared with the normal group, the expression of rno-miR-21-5p, rno-let-7i-5p, rno-miR-146b-5p and rno-miR-15b-5p in model group increased significantly(PPPPPConclusion: This experiment found 4 miRNAs involved in the regulation of renal interstitial fibrosis (RIF) and 2miRNAs with unknown functions, which provided a new clue for further analysis of the regulatory network of kidney-yang deficiency.

Effect of Zhenwu Tang on regulating of "AVP-V2R-AQP2" pathway in NRK-52E cells / 中国中药杂志

This study was aimed to investigate the effect and mechanism of Zhenwu Tang on AVP-V2R-AQP2 pathway in NRK-52E cells . Forty eight male SD rats were randomly divided into eight groups with 6 animals in each group. Distilled water or 22.68 g·kg⁻¹·d⁻¹ Zhenwu Tang(calculated by raw drug dosage meter) was given by gavage. Blood samples were collected by cardiac puncture， and the medicated serum was centrifuged from the blood by 3 000 r·min⁻¹. NRK-52E cells were treated with different medicated serum or dDAVP. The condition of cell proliferation was detected by RTCA. The distribution of V2R and AQP2 in cells were detected by immunofluorescence. The expression of V2R， PKA and AQP2 were detected by Western blot and AQP2 mRNA level was detected by real-time PCR. Results showed that the level of AQP2 mRNA(<0.01) and protein expression of V2R， PKA and AQP2(<0.05， <0.01， <0.05) of Z7d group which was treated with Zhenwu Tang medicated serum for 24 h were significantly higher than that of normal rat serum group. And the expression level of V2R， p-AQP2 and AQP2(<0.01， <0.05， <0.01) of Z7d+dDAVP group were significantly increased comparing to normal rat serum group. The results indicate that the applying of Zhenwu Tang medicated serum could increase the expression level of V2R， PKA and AQP2 which exist in AVP-V2R-AQP2 pathway in NRK-52E， and there is synergistic effect between Zhenwu Tang medicated serum and dDAVP. So the pathway of AVP-V2R-AQP2 may be one of the mechanism for which Zhenwu Tang regulate balance of water transportation.

Protective effect of β-asarone on AD rat model induced by intracerebroventricular injection of Aβ₁₋₄₂ combined 2-VO and its mechanism / 中国中药杂志

This study was aimed to investigate the protective effect and mechanism of β-asarone on the animal model of Alzheimer's disease(AD) which was induced by intracerebroventricular injection of Aβ₁₋₄₂ combined cerebral ischemia. One hundred and five rats were randomly divided into seven groups including sham-operated group, AD model group, β-asarone 10 mg•kg⁻¹ group, β-asarone 20 mg•kg⁻¹ group, β-asarone 30 mg•kg⁻¹ group, donepezil group(0.75 mg•kg⁻¹) and Ginkgo biloba extract group(24 mg•kg⁻¹). Rats' learning and memory abilities, cerebric regional blood flow, pathological changes in hippocampal CA1 region, the expression level of HIF-1α and serum CAT, SOD and MDA level were detected 4 weeks later. The results showed that the application of intracerebroventricular injection of Aβ₁₋₄₂ joint 2-VO could lead to rats' dysfunction of learning and memory, decrease in regional cerebral blood flow. Neurons in CA1 region were arranged in disorder, and amyloid deposition was increased. The number of cerebral cortical cells expressing HIF-1α was increased as well. The level of serum CAT and SOD decreased, while level of serum MDA increased. However these symptoms were improved by 20 mg•kg⁻¹ and 30 mg•kg⁻¹ β-asarone. The results indicated that β-asarone could effectively relieve the symptoms of the AD model induced by intracerebroventricular injection of Aβ₁₋₄₂ combined cerebral ischemia, and the potential mechanism might be that it could attenuate damage of MDA to the body by improving the level of CAT and SOD, meanwhile the level of HIF-1α decreased as the decline of hyperoxide which might attenuate its damage to neuron, so it finally achieved alleviating Alzheimer's disease.

Protective effect of β-asarone on PC12 cells injury induced by Aβ₁₋₄₂ astrocytic activation / 中国中药杂志

This study was aimed to investigate the protective effect and mechanism of β-asarone on PC12 cells injury induced byAβ₁₋₄₂ activated astrocytes, and provide experimental basis for β-asarone application in the prevention and control of Alzheimer's disease (AD). Firstly, RA-h and PC12 cells were co-cultured in the special transwell chamber, and the Real time cell analysis (RTCA) system was used to real-time observe its effect on PC12 cells survival rate in the co-culture system after astrocytes injury induced by Aβ₁₋₄₂. The best intervention time of β-asarone was selected according to the survival curve and parameters generated automatically. β-asarone with different concentrations was used for intervention on astrocytes, then the changes of PC12 cells survival rate in the co-culture system were observed. Secondly, MTT assay was used to detect the effect of Aβ₁₋₄₂ on PC12 cells survival rate as well as the intervention effect of β-asarone, and verify the testing results of RTCA. The levels of IL-1β, TNF-α and BDNF in culture media of the lower chamber were detected by ELISA. The NF-κB activity and phosphorylation levels of ERK, p38 and JNK were detected by Western blot. Results showed that β-asarone (55.5 mg•L⁻¹) could significantly slowdown the decline of PC12 cells survival rate caused by Aβ₁₋₄₂-induced RA-h activation (P<0.01), significantly reduce the levels of IL-1β, TNF-α and the phosphorylation levels of ERK, p38 and JNK in culture media of the lower chamber (P<0.01). β-asarone(166.7 mg•L⁻¹) could promote the release of BDNF in culture media of the lower chamber(P<0.05). These results indicated that Aβ₁₋₄₂ could induce RA-h activation and its release of IL-1β, TNF-α and other inflammatory factors to aggravate the PC12 cells injury; β-asarone could reduce the levels of IL-1β, TNF-α, promote the release of BDNF, and inhibit the NF-κB activity as well as phosphorylation levels of ERK, p38 and JNK protein in PC12 cells.

Identification of the active material of anti-hepatic fibrosis from Amydae Carapax / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To identify the active material of anti-hepatic fibrosis from Amydae Carapax.</p><p><b>METHODS</b>Membrane separation technology was adopted to screen active fraction in Amydae Carapax, and the active components were isolated from the active fraction using gel chromatography and high performance liquid chromatography. The purified active components in Amydae Carapax were further analyzed using 4700 series time-of-flight mass spectrometer.</p><p><b>RESULTS</b>Proteins and peptides of Amydae Carapax with molecular weight less than 6000 were proved to have biological activity. 8 components (Bj1-Bj8) were isolated from the active fraction. Bj4, Bj6 and Bj7 were screened as active components. Bj7 was further purified, resulting in 7 components (Bj701-Bj707). Bj704 and Bj707 showed significant biological activity. Mass spectrometry showed three molecular ion peaks with highest abundance, i.e. m/e 526, 542 and 572, i.e. m/e 526, 542 and 572, in Bj707 -A The amino acid sequences of above three peptide compounds were NDDY (Asn-Asp-Asp-Tyr), NPNPT (Asn-Pro-Asn-Pro-Thr), and HGRFG (His-Gly-Arg-Phe-Gly), respectively. And M572 was the most abandunt components.</p><p><b>CONCLUSION</b>Three active peptide compounds of anti-hepatic fibrosis of Amydae Carapax were identified.</p>

Comparison of the distribution of breviscapine in the brain by different administration routes / 药学学报

By comparing the drug distribution of breviscapine administered intranasally, orally and intrgvenous injected in rats' brain. After 0.4 mg x kg(-1) breviscapine was given by tail vein, intranasal and gastric perfusion administration to SD rats, cerebrospinal fluid was obtained by erebellomedllery cisternal puncture at different times. 125I labeling was used to determine the drug content of cerebrospinal fluid, cerebrum, cerebellum, medulla oblongata, olfactory region, olfactory bulb and blood in rats. AUCs were calculated by trapezoidal rule. The results showed that AUCs(0-240 min) (microg x min x g(-1)) of brain tissues were 11.686 +/- 1.919, 5.676 +/- 1.025, 7.989 +/- 0.925, 7.956 +/- 1.159, 17.465 +/- 2.136, 24.2 +/- 2.906 and 78.51 +/- 12.05, respectively, in the intranasal administration group; while those in the tail vein administration groups were 6.79 +/- 0.661, 6.251 +/- 0.40, 10.805 +/- 1.161, 9.146 +/- 1.04, 9.892 +/- 1.532, 7.871 +/- 0.842 and 173.91 +/- 10.02; and oral administration group were 0.868 +/- 0.167, 1.708 +/- 0.266, 2.867 +/- 0.725, 2.067 +/- 0.313, 1.361 +/- 0.308, 1.206 +/- 0.255 and 45.2 +/- 7.52, respectively. AUCs(0-240 min) of the brain tissues after oral, tail vein and intranasal administration were 22.29%, 29.18%, 95.49% of that of blood, respectively, it means that the absorption rate and drug distribution in the brain tissues after intranasal administration were higher than those of oral and tail vein administration. It is worth to investigate further the pharmacodynamic relationship.

Adverse drug reactions induced by Mailuoning injection: literature review on 162 cases / 中国中药杂志

To understand the adverse drug reaction (ADR) induced by Mailuoning injection. 162 ADRs due to the drug were retrieved from national medical journals of 1988-2007 for statistics. It was shown that there was no relationship between ADR and dosage, but ADR appeared mostly in middle-aged and old groups, and more in male than in female. The occurrence of ADR was commonly within 30 min after injection. It involved injuries of various systems and organs. As for the 123 cases of allergy, 38 were anaphylactic shock (accounting for 23.46%), two people died. ADR characterized by immediate type on initial use and tachy type. It should be paid more attention to the Mailuoning injection for the immediate hypersensitivity reaction (such as anaphylactic shock).

The changes of three endogenous hormones during flower bud differentiation of Schisandga chinensis / 中国中药杂志

<p><b>OBJECTIVE</b>To reveal the relation between endogenous hormones and the flower bud differentiation in Schisandga chinensis.</p><p><b>METHOD</b>Top buds of extremely short branch and axillary buds of long branch in the same plant of S. chinensis were used as material and the contents of endogenous hormones were measured during different periods of the flower bud differentiation with HPLC.</p><p><b>RESULT</b>The result showed that flower bud differentiation and the formation of female flower were inhibited by high concentration of GA3 and were promoted by high concentration of ABA or ZT. Low ratio of GA3/ABA has the same result.</p><p><b>CONCLUSION</b>There was a correlation between endogenous hormones and the flower bud differentiation of S. chinensis.</p>

Experimental study of traditional Chinese medicine in the treatment of vascular dementia / 中国中药杂志

To explore the new progress of the experimental study of traditional Chinese medicine in the treatment of vascular. Consulted the relevant papers and integrated the elementary theory, we summarized the virtue of Chinese medicine in the treatment of vascular dementia and insufficient and future development of the experimental study of it. It was confirmed that the traditional Chinese medicine has a lot of target spots in the brain and less ill-effect, it could ameliorate several phenotypes of pathophysiology of vascular dementia and improve the ability of learning and memory of animals with vascular dementia.

Induction of hairy roots of Panax ginseng and studies on suitable culture condition of ginseng hairy roots / 生物工程学报

Ginseng is a valuable medicinal plant with ginsenosides as its mian effective components. Because ginseng is a perennial plant and has a very strict demand for soil conditions, the way of cultivating ginseng by cutting woods is still used in China at present and thus forest resources has been extremely destroyed. Increasing attention has been paid to the hairy roots induced by the infection of Agrobacterium rhizogenes in the production of plant secondary metabolic products for the hairy roots are characterized by rapid growth and stable hereditary and biochemical traits. That has opened a new way for the industrial production of ginseosides. However, there is little report for such studies from China. In this paper, hairy roots of ginseng were induced from the root explants of two-year-old ginseng by Agrobacterium rhizogenes A4 with directly inoculating. The transformed hairy roots could grow rapidly on MS medium and 1/2 MS medium without hormones. The cultured clones of the hairy roots were established on a solid 1/2 MS medium. After 4 - 5 subcultures the hairy roots still maintained a vigorous growth. A pair of primers were designed and synthesized according to the analytical results of RiA4TL-DNA sequence by Slightom et al . 0.8kb rolC was obtained by PCR using the genome DNA of hairy root of ginseng. Transformation was confirmed by PCR amplification of rolC genes from the hairy roots of P. ginseng. Growth rate of hairy roots on liquid medium increased by 2 times then that of the solid medium. The growth of the hairy roots can be divided into three stages: high speed in the first two weeks, middle speed in the 3 - 4 weeks and low speed hereafter. Changing the culture solution at 2 weeks regular intervals is conductive to maintaining the rapid growth of the hairy roots. By means of determination for specific growth rate and ginsenosides content, the high-yield hairy root clone R9923 was selected. The content of monomer gisenoside of Rg1, Re, Rf, Rbl, Rc, Rb2 and Rd in hairy root clone R9923 was determined by the HPLC. The total ginsenosides content in the hairy toot clone R9923 came up to 15.2 mg/g. The suitable culture conditions for ginseng hairy roots growing were 1/2 MS liquid medium (30 g/L glucose), in a shaker at 110 r/min, changing the culture solution at 2 weeks and subculture time 4 weeks. In the liquid fermented culture of 2L medium, the yield of the hairy roots could amount to 270.10 g in 4 weeks. The industrial production of ginsenosides has been preliminarily realized. Effect factors on biomass and ginsenosides content such as culture volume, inoculation, in steps cultural technology at the scale-up process of hairy roots culture were also explorated. Our results have laid a foundation for defining optimum culture manner for large-scale cultivation and large-scale production of ginsenosides.

Antioxidative effect of total axtract of astragalosides / 中国临床药理学与治疗学

Aim The relationship between the therapeutic effect of TEA (total extract of astragalosides) on adjuvant arthritic (AA) rats and its antioxidative effects were studied . Methods The volume of non-injected hind paw of AA rats and malondialdehyde(MAD) content of arthritic synoviocytes from AA rats were measured and the proliferative responses of fibroblast, the level of superoxide anion() and the hydroxyl radical (?OH) generated in vitro were detected . Results The anti-inflammatory effect of TEA might be related to its antioxidative activity.In vitro low-level oxidative stress promoted the proliferative responses of fibroblast in rats synovium, which was marked by inhibited by TEA in a concentration-dependant manner. Further study showed that TEA could inhibit NBT reduction induced by both xanthine-xanthine oxidase and non-enzyme generated , but the inhibitory effect of this compound on activity of xanthine oxidase was obtained only at high concentration (more than 80 ?g?ml-1). It was also found that TEA could dose-dependantly inhibit the hydroxylation of benzoic acid induced by Fenton reaction generated ?OH. Conclusion TEA has significant therapeutic effects on AA rats, which might be related to its antioxidative effects