AIM To establish an HPLC method for the simultaneous content determination of saikosaponin a,saikosaponin c,saikosaponin d,rutin,quercetin,isorhmnetin and kaempferol in the aerial and underground parts of “Zhongchai No.2” (Bupleurum chinense DC.from Sichuan Province).METHODS The analysis of “Zhongchai No.2” methanol extract was performed on a 30 ℃ thermostatic Hypersil GOLD column (250 mm × 4.6mm,5.0 μm),with the mobile phase comprising of acetonitrile-0.1％ phosphoric acid flowing at 0.8 mL/min in a gradient elution manner,and the detection wavelength was set at 210 nm.RESULTS Seven constituents showed good linear relationships within their own ranges (r≥0.999 1),whose average recoveries were 96.30％-102.7％ with the RSDs of 0.95％-3.8％.There were a lot of flavonoids in the aerial parts of samples,while a large number of saponins existed in the underground parts,but almost no flavonoids were found.CONCLUSION The aerial parts (stems and leaves) of “Zhongchai No.2” also have medicinal value,to which we should pay attention.

OBJECTIVETo investigate the inhibitory effects of saikosaponin-d (SSd) on angiogenesis in chicken embryos and its mechanism of action.

METHODSChorioallantoic membrane (CAM) model was established successfully in 86 chicken embryos. They were divided into 4 groups after fenestration: the three SSd treated groups (A, B and C) treated with high (20 microg/mL, n = 16), middle (10 microg/mL, n = 19) and low (5 microg/mL, n = 25) dose of SSd respectively, and the control group treated with 0.01 mol/L PBS (n = 26). The drug or reagent was administered by grafting 20 microL onto the surface of CAM. After incubation for 3 days, the vessel growth was recorded by digital photography; inflammatory cells were counted under light microscope with HE staining, and the positive rate of angiogenesis reaction was calculated by Leica image analyzer.

RESULTSOn the 6th day of the embryonic age, vessels in the chicken embryo CAM showed a radial growing in spok-wheel pattern around the gelatin sponges with lateral axis running through it. Whereas after 3 days of SSd treatment, the angiogenesis reduced significantly with vague microvessels around the sponge, and vascular truncation and absence revealed. Microscopic examinations showed that the number of microvessels and infiltrated inflammatory cells in the sponge and peripheral CAM mesenchyme in the SSd groups were less than those in the control group, especially on vessels of medium and small size (P < 0.05, P < 0.01, respectively), but was insignificant on great vessels (P > 0.05). Correlation analysis revealed no correlation between the number of the great vessels in CAM and the infiltrated inflammatory degrees (r = 0.117, P > 0.05), but the increase of small vessels in CAM was positively correlated with that of inflammatory cells (r = 0.971, P < 0.01).

CONCLUSIONSSSd could inhibit the physiological angiogenesis of chicken embryoe, especially for the medium and small vessels, while there was no significant effect on great vessels (P > 0.05). Its mechanism of action may be related to its inhibition on leukocyte migration and activation.

Angiogenesis Inhibitors ; pharmacology ; Animals ; Chick Embryo ; Chorioallantoic Membrane ; drug effects ; Neovascularization, Physiologic ; drug effects ; Oleanolic Acid ; analogs & derivatives ; pharmacology ; Saponins ; pharmacology

OBJECTIVETo confirm the amount of fertilizer and the ration of fertilizer.

METHODThrough the experiment of planting Salvia miltiorrhiza in pot and in field were carried out.

RESULTWhen N:P = 1:1, The production was 1.8 times more than the contrast group in plotting experiment, and in field the production of the highest fertilizer plot were 2.5 times more than the contrast and the higher and high fertilizer plot is 2.25 and 1.2 times respective were than those in the contrast group.

CONCLUSIONThe proper ration of nitrogenous fertilizer and phosphorous fertilizer is 1:1. Nitrogen shows negative effects to the accumulation of tanshinon IIA, the more the nitrogen ous fertilizer, The less the content of tanshinon IIA. In contrast, phosphor ous fertilizer shows good effects on the accumulation of tanshinon IIA. Phosphorous fertilizer could alleviate the decline of the content of tanshinon IIA by using nitrogenous fertilizer. The accumulation peak of the tanshinon IIA emerge in the period of 150 d.

Diterpenes, Abietane ; Fertilizers ; Nitrogen ; analysis ; pharmacology ; Phenanthrenes ; analysis ; Phosphorus ; analysis ; pharmacology ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; growth & development ; Salvia miltiorrhiza ; chemistry ; growth & development

The pediatric population is different from adults in their physi-ological function and tissues structure, and therefore they should not be assumed simply as the small adults.The drug development for pediatric population is helpful to improve the health of children in the whole coun-try.However, the clinical investigation of medicinal products for adults can not apply for children and there has not available guidance for chil-dren for a while.Accordingly, Chinese Food and Drug Administration ( CFDA) issues the guidance for clinical investigation of medicinal prod-ucts in the pediatric population, which ensure the clinical trials are more scientific and ethic. This paper means to interpret the guidance and therefore to help the development of clinical trials in pediatric population.

The pediatric population is different from adults in their dis-ease types and medication use in clinics.The drug clinic trial in pediatric population provides reliable clinic data for new drug development and medication use, and is helpful to promote the health of children.Howev-er, considering the specificity of children, it is changeable to conduct the drug clinic trials, especially for the study protocol design.Accordingly, CFDA issues the guidance for clinical investigation of medicinal products in the pediatric population, which ensure the clinical trials are more sci-entific and ethic.This paper means to interpret the guidance and there-fore to help the development of clinical trials in pediatric population.

OBJECTIVETo explore the inhibitory effect of estrogen against metastasis of human hepatocellular carcinoma MHCC97H cells and explore the molecular mechanism.

METHODSThe inhibitory effect of estrogen on the migration and invasion of MHCC97H cells was evaluated with wound healing assay and Transwell assay. Western blotting was used for investigating the expression of MMP-2, MMP-9, AKT and p-AKT in the cells treated with estrogen.

RESULTSEstrogen treatment significantly inhibited the migration and invasion of MHCC97H cells in a dose-dependent manner. Estrogen significantly down-regulated the protein expressions of MMP-2 and MMP-9 and lowered the phosphorylation level of AKT.

CONCLUSIONThe anti-metastatic effect of estrogen involves inhibition of MMP-2 and MMP-9 in MHCC97H cells probably by regulating AKT signal pathway.

BACKGROUNDChinese medicine plays an important role in hepatoprotective treatment. This study was conducted to investigate the protective effects of emodin and astragalus polysaccharides (APS) in a rat model of chronic hepatic injury.

METHODSChronic hepatic injury was induced by hypodermic injection of an olive oil solution containing 40% carbon tetrachloride (CCl(4)) twice a week, in addition to a diet of 79.5% maizena, 20% fat, 0.5% cholesterol, and 10% alcohol in the drinking water ad libitum for 12 weeks. Meanwhile, the rats were exposed to different concentrations of emodin (40 mg x kg(-1) x d(-1)), APS (200 mg x kg(-1) x d(-1)), combination drug (emodin 40 mg x kg(-1) x d(-1) combined with APS 200 mg x kg(-1) x d(-1)) and colchicine (0.1 mg x kg(-1) x d(-1)) in parallel by oral gavage (once a day for 12 weeks). At the end of 12 weeks, blood serum and liver tissue were taken. Serum was collected to determine the levels of total bilirubin (TBIL), alanine transaminase (ALT), aspartate transaminose (AST), and albumin (ALB). Liver and spleen indexes were assayed, followed by the measurements of the liver associated enzyme superoxide dismutase (SOD) and malondialdehyde (MDA). Histopathological changes were studied using optical microscopy.

RESULTSSplenohepatomegalia was alleviated and serum levels of TBIL and ALT were reduced in the groups treated with emodin and APS when compared to the control group. In addition, the ALB level in the APS and combination groups was higher. Similarly, the SOD activity of liver homogenates was significantly higher in the groups treated with emodin and APS, while administration of the herbal derivatives prevented the elevation in MDA levels. Histological analysis showed that the APS and combination groups significantly ameliorated the hepatic injury.

CONCLUSIONSCo-administration of emodin and APS demonstrated a synergistic action in reducing ALT and restoring ALB in the serum from a rat model of chronic hepatic injury. Emodin and APS may ameliorate the CCl(4)-induced hepatic injury in rats by elevating antioxidant-enzyme activities and reducing lipid peroxidation.

Alanine Transaminase ; blood ; Animals ; Astragalus Plant ; chemistry ; Carbon Tetrachloride ; toxicity ; Chronic Disease ; Emodin ; pharmacology ; Liver ; drug effects ; pathology ; Male ; Malondialdehyde ; analysis ; Polysaccharides ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

BACKGROUNDHuman urate anion exchanger (hURAT1) as a major urate transporter expressed on renal tubular epithelial cells regulates blood urate level by reabsorbing uric acid. Antibody is an important tool to study hURAT1. This study aimed, by genetic immunization, to produce mouse anti-hURAT1 polyclonal antibody with high throughput and high specificity and to detect the location of hURAT1 in human kidney.

METHODSHuman renal total RNA was isolated and the entire cDNA of hURAT1 was amplified by RT-PCR. The sequence of intracellular high antigenicity fragment (A280 to R349) was chosen by prediction software of protein antigenicity, and its cDNA was amplified from cDNA of hURAT1, and then cloned into pBQAP-TT vector to construct recombinant plasmid pBQAP-TT-hURAT1-210 for genetic immunization. Mice were inoculated with this recombinant plasmid and two other adjuvant plasmids, pCMVi-GMCSF and pCMVi-Flt3L, which helped to enhance the antibody's generation. After four weeks, the mice were sacrificed to obtain the anti-hURAT1 antibody from serum. The antibody was identified by western blot analysis and immunohistochemistry. At the same time, rabbit anti-hURAT1 antibody was produced by protein immunization. The specificity and efficiency between the rabbit and mouse anti-hURAT1 antibody were compared by western blot analysis and immunohistochemistry.

RESULTSThe entire cDNA of hURAT1 and cDNA of its intracellular high immunogenic fragment were amplified successfully. Recombinant plasmid pBQAP-TT-hURAT1-210 for genetic immunization was confirmed by restriction digestion and sequencing. Both the mouse anti-hURAT1 antibody and rabbit anti-hURAT1 antibody recognized 58 kD hURAT1 and 64 kD glycosylated hURAT1 protein bands in western blot. Immunohistochemically, hURAT1 was located at the brush border membrane of renal proximal tubular cells. In addition, the throughput and specificity of the mouse anti-hURAT1 antibody were higher than those of the rabbit anti-hURAT1 antibody.

CONCLUSIONGenetic immunization can generate anti-hURAT1 polyclonal antibody of high throughput and specificity.

Animals ; Antibodies ; analysis ; Blotting, Western ; Carrier Proteins ; analysis ; immunology ; Female ; Humans ; Immunization ; Immunohistochemistry ; Kidney ; chemistry ; Male ; Mice ; Organic Anion Transporters ; analysis ; immunology ; Organic Cation Transport Proteins ; Plasmids ; Rabbits

OBJECTIVETo observe the therapeutic effect of acupoint sticking therapy for treatment of stroke.

METHODSTwo hundred and forty-six cases were randomly divided into a non-acupoint sticking group (control group, n=122) and an acupoint sticking group (n=124). The control group was treated with routine ward treatment of stroke (acupuncture combined with routine western medicine). The acupoint sticking group was treated with basis ward treatment of stroke (similar to the control group), and acupoint sticking therapy was applicated on Shenque (CV 8). The scores of Stroke-Specific Quality of Life (SS-QOL) and WHOQOL-100BREF were adopted to evaluate the therapeutic effects and the cysteine of patients were tested before and after treatment.

RESULTSThere were significant differences in scores comparison of SS-QOL and WHOQOL-100BREF before and after treatment in both groups (both P < 0.001); there was no significant difference after treatment between two groups (P > 0.05); there was a significant difference in thinking items of SS-QOL after treatment between two groups (P < 0.05), the acupoint sticking group was superior to that of control group; SS-QOL score of patients with abnormal cysteine of acupoint sticking group was superior to that of the control group after treatment, with a significant difference between the two groups (P < 0.05).

CONCLUSIONThe acupoint sticking therapy can improve the thinking ability of stroke patients, and improve the life quality of high cysteine stroke patients.

Acupuncture Points ; Acupuncture Therapy ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Stroke ; psychology ; therapy ; Thinking

Objective:In the early stage, in vivo experiments of coronavirus infection and pulmonary syndrome syndrome combined with mouse model verified that Reyanning mixture has a significant therapeutic effect on human coronavirus disease-2019 (COVID-19). This study further evaluated the clinical efficacy of Reyanning mixture in the treatment of COVID-19, providing a basis for clinical medication. Method:Patients were collected from January 21, 2020 to February 24, 2020 in Xi'an No. 8 Hospital, Second Affiliated Hospital of Xi'an Jiaotong University, Yan'an Second People's Hospital and Songzi People's Hospital. 54 common-type patients who met the diagnostic criteria of COVID-19 were enrolled in this study,5 patients of them were not included in the statistical analysis because they did not meet the requirements, including 26 cases in the treatment group using Reyanning mixture on the basis of chemical drug treatment, and 23 cases in the control group receiving chemical drug treatment only. The symptom disappearance rates (throat dryness, throat pain, cough, fever, fatigue, chest tightness, runny nose, nasal congestion, headache), time to complete fever clearance (d), the nucleic acid conversion rate and time to recovery on chest CT were compared between two groups. Result:After treatment, except cough and fatigue, other symptoms disappeared in the treatment group, and the disappearance rate of symptoms such as dry throat, cough, fatigue, chest tightness and headache was statistically significant compared with the control group (P<0.05). The median time to complete fever clearance in the treatment group was 3 days, which was 2 days shorter than that in control group. The remission rate of chest CT was 88.46% (23/26) in the treatment group, which was higher than 73.91% (17/23) in control group. The negative conversion rate of viral nucleic acid detection was 96.15% (25/26) in treatment group, higher than 60.87% (14/23) in control group (P<0.01). Conclusion:Reyanning mixture can improve the clinical symptoms of COVID-19 patients, promote the improvement of chest CT. Can shorten the duration of fever.Can improve the novel coronavirus nucleic acid conversion rate, providing a basis for clinical treatment.