This paper reported a new method of Armillaria mellea isolation-Gastrodia elata tissue isolating. Compared with normal isolating method-rhizomorph isolating method, it showed that the success rate of new method (78% ) was higher than the rhizomorph isolating method (16% ) , besides this, the new method was easier, and growth characteristic of obtained strain was superior to that obtained from rhizomorph isolating method.

AIM: To explore whether strontium ranelate ( Sr ) promotes osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs) through the Hedgehog/Gli1 signaling pathway.METHODS:BMSCs were isolated from 4-week-old rats by adherent culture and induced to differentiate into osteoblasts .According to the experimental purposes , the cells were exposed to different concentrations of Sr , cyclopamine ( Cy, an inhibitor of Hedgehog receptor ) or Gli1-siR-NA.The expression of Gli1 and Runx2 in the cells was detected by Western blotting .The activity of alkaline phosphatase ( ALP) was measured by the method of colorimetry , and the mineralized nodules were observed under microscope with aliz-arin red staining .RESULTS:Exposure to Sr at concentrations of 0.1 to 5 mmol/L for 7 d markedly increased the expres-sion of Gli1 in the BMSCs , and the increase in Gli1 expression was the most obvious following Sr exposure at concentration of 3 mmol/L.Cy at concentration of 10 μmol/L inhibited Sr-induced up-regulation of Gli1 expression.Transfection of the BMSCs with Gli1-siRNA not only obviously inhibited Sr-induced up-regulation of Gli1 and Runx2 ( a downstream protein of Gli1) expression, but also antagonized Sr-induced enhancement of ALP activity and the formation of mineralized nodules . CONCLUSION:The Hedgehog/Gli1 pathway is involved in Sr-induced osteogenic differentiation of rat BMSCs .

To study the chemical components and the antifungal activity of extraction from conidia of Trichoderma viride LTR-2.The extraction were obtained by distilling with Methylene dichloride from conidia of Trichoderma viride LTR-2 cultured on wheat bran solid matrix.Antifungal activity were determined by mycelium growth method.The chemical components of the extraction were analysed by GC-MS,the relative components in the extraction were determined by area normalization.The extraction not only have broad-spectrum control,showed antibiosis against eleven different plant fungal pathogens in PDA dish,such as Rhizoctonia solani,Alternaria brassica,Verticillium dahliae,Macrophoma kawatsukai,Fusarium moniliforme,Botrytis cinerea,Rhizoctonia cerealis,Fusarium oxysporum f.sp.vasinfectum,Bipolaris sorokinana,Fusarium graminearum,Alternaria.mali,but also have high inhibitory effect,and had 89.3% suppressive rate to Rhizoctonia cerealis.About sixty components were separated and identified by GC-MS,majority components were Hydrocarbon,the number of the Hydrocarbon were fourty-three kinds.Ergosterol was the major chemical components of the extract,and has 41.90% content.Other components comprised:Ketone,Organic acid,Alcohol,Ene,et al.Conclusion:The extraction from conidia of Trichoderma viride LTR-2 have antifungal activity.The extration comprised 2H-Pyran-2-one,5,6-dihydro-6-pentyl,it has 2.35% content.reference others literature,2H-Pyran-2-one,5,6-dihydro-6-pentyl may be the suppressive component of the extration.

Objective To study the influence and dose effect of ultrasound on the contraction of uterine smooth muscle in rats.Methods Estradiol benzoate was injected into rats three days before conducting an in-vitro experiment.Their uteri were resected and irradiated with ultrasound(0.8 MHz,3 W/cm~2,0-40 rain).The contrac- tion frequency and amplitude were recorded using an MS-302 biological experiment system.Results It could be seen that the contraction frequency and amplitude,and general contractile activity were significantly increased during ultrasonic irradiation(P＜0.01).The increased contraction frequency and amplitude lasted for ten minutes,and then the normal contraction pattern resumed.The contraction frequency as well as the percentage change in eontraction fre- quency were highest during the first 15 minutes of ultrasonic irradiation;the contraction amplitude as well as the per- centage change in amplitude were highest during 40 minutes of ultrasonic irradiation.Contraction activity was at its highest for 30 minutes,but the percentage change in activity was highest for 20 minutes.Conclusions Ultrasound can induce uterine smooth muscle contraction in rats.This biological effect is related to the irradiation time.

OBJECTIVETo study the post-operative pain-re1ated factors in lower third molar extraction, and to find an effective prevention program.

METHODSThe first phase of the study was a clinical follow-up of 450 cases of lower third molar extraction. Factors associated with post-operative pain were analyzed, and difficulty assessment protocol of the tooth extinction was build. On this hasis, a pain-prevention clinical plan was constitute, and implemented in the second phase of 450 cases (selectively systemic or local medication). Treatment effects were observed.

RESULTSThere were no significant differences between the two phase groups oh the distribution of ages, sex, difficulty of tooth extraction, operation time, and other related factors, means that they were statistically comparable. Tooth extraction difficulty and operation time were most important predictive factors for post-operative pain. Local infective factors (including caries) and women's menstrual cycle also account for late stage pain. Through targeted pre-operative medication or post-operative alveolar socket treatment, pain was significantly reduced or prevented.

CONCLUSIONReasonable peri-operative drug control basing on the difficulty assessment can provide a comprehensive and individualized control of pain after tooth extraction. Also, excessive or inappropriate using of drugs is largely avoided.

Adult ; Female ; Humans ; Molar ; Molar, Third ; Pain ; Tooth Extraction

OBJECTIVEDiet is an important factor influencing blood pressure and, increases in dietary carbohydrate intake can raise blood pressure in adult rats. A previous study showed that the blood pressure of the rats fed with high-carbohydrate was 5-20 mmHg higher than that of control rats. While the mechanism involved is not clear. This study aimed to investigate the effects of high-sucrose intake on blood pressure of young Wistar rats and the role that sympathetic nerve system in the process.

METHODSMale neonatal Wistar rats were performed sympathectomy operation with 6-hydroxydopamine (6-OHDA) and then divided into four groups: (1) 0.1% VitC saline-common diet group (VN), (2) 0.1% VitC saline-high sucrose (VS), (3) 6-OHDA-common diet group (OHN) and (4) 6-OHDA-high sucrose (OHS) after three week. The data on the body weight (BW), systolic blood pressure (SBP) and heart rate (HR) were recorded. Then the level of blood glucose, serum insulin and angiotensin II (AngII) were measured and the functional studies of the thoracic aorta was performed.

RESULTSThe VS group exhibited higher SBP than the OHS group from the 6th week (113.7 +/- 4.2 mmHg vs. 104.0 +/- 5.8 mmHg, P < 0.01) and the VN group from the 7th week (117.6 +/- 6.3 mmHg vs. 109.6 +/- 4.6 mmHg, P < 0.01), while the SBP of the VN group was similar to those of the OHN group and the OHS group (P > 0.05). No significant differences in blood glucose, serum insulin and insulin sensitive index (ISI) were found among the four groups. The thoracic aorta segments of the VS group had higher contractive response to AngII (P < 0.01) and NE (P < 0.05) than the VN group, but the relaxations to acetylcholine (ACh) and nitroglycerine (NTG) showed no difference among the four groups (P > 0.05).

CONCLUSIONThe high-sucrose diet might elevate the blood pressure in young Wistar rats and the sympathetic system may play an important role in this process.

Angiotensin II ; blood ; Animal Feed ; Animals ; Blood Glucose ; metabolism ; Blood Pressure ; drug effects ; physiology ; Body Weight ; drug effects ; Dietary Sucrose ; administration & dosage ; Insulin ; blood ; Male ; Oxidopamine ; administration & dosage ; blood ; Rats ; Rats, Wistar ; Sympathectomy

In order to establish the stable andreliable ISSR-PCR System of Lysimachia christinae, L16 (4(5)) orthogonal design, which based on 7 levels of single factor experiment, were used in this study. The variance analysis was carried out by SPSS 19.0, and 5 main factors affecting the reaction system were optimized in 4 levels. The best annealing temperature was selected by the optimized reaction system. And the stability and reliability of this system was tested by 23 samples from different origins. The results showed that the five factors (DNA template, primer, dNTP, Mg2+ and Taq enzyme) were the most impacts on the amplified results of ISSR-PCR of L. christinae. The order of the influence was: primer > Taq enzyme > DNA template > Mg2+ > dNTP. The optimal system, which was determined by multiple comparison on different levels of each factor, was total volume of 25 microL, including DNA template 60 ng, primer 0.3 micromol x L(-1), dNTP 0.2 mmol x L(-1), Mg2+ 1.8 mmol x L(-1), Taq enzyme 1.25 U. The optimal system was stable and reliable tested by 23 samples from different origins. This study lays the foundation for genetic diversity analysis, fine varieties selection and molecular identification of L. christinae, and provides reference for optimization on ISSR-PCR system of other speciesin future.
DNA Primers ; genetics ; DNA, Plant ; genetics ; Drugs, Chinese Herbal ; chemistry ; classification ; Microsatellite Repeats ; Polymerase Chain Reaction ; methods ; Primulaceae ; classification ; genetics ; Quality Control

OBJECTIVETo improve the design of the devices used for autoatic elasticity distraction osteogenesis, and to preliminarily testify their feasibility for clinical application.

METHODSThe internal automatic distraction devices were constructed by three parts, including sinusoid shape memory alloy distractor, reconstructive plate and mini-plate. The animal model of trifocal bone transport reconstruction of canine mandibular mentis was duplicated, in which a 6 cm mandibular mentis segment was removed and the transport disc was created by osteotomy in both ends of the mandibular defect. The plates were used for internal fixation and shape reconstruction of the mandible. They also served as transport distraction guidance for the elasticity distractor.

RESULTSThe new devices could complete bone transport as expected. Regenerated bone segments were formed in two months and both sides of the mandibular body were lengthened. Because of the narrow angle and obstruction from the thick soft tissues in the mentis area, the ultimate symphysis mentalis was not reconstructed.

CONCLUSIONThe design of the internal elasticity distraction devices showed potential feasibility for clinical practice. But further studies are still needed for better control of the strength and balance of distraction forces.

Animals ; Bone Regeneration ; Elasticity ; Mandible ; Osteogenesis, Distraction ; Reconstructive Surgical Procedures

Adult ; Aged ; Bone Transplantation ; Female ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Thoracic Vertebrae ; injuries ; surgery

BACKGROUND: There is growing evidence that 5-fluorouracil (5-FU) combined with therapeutic trauma can effectively induce skin repigmentation in vitiligo patients who are unresponsive to conventional treatments. Previous studies have mainly focused on identifying the antimitotic activity of 5-FU for the treatment of skin cancer, but few studies have investigated its extra-genotoxic actions favoring melanocyte recruitment. METHODS: We utilized the full thickness excisional skin wound model in Dct-LacZ transgenic mice to dynamically assess the migration of melanocytes in the margins of wounds treated with or without 5-FU. The in-situ expression of CXCL12 was examined in the wound beds using immunofluorescence staining. Quantitative real-time polymerase chain reaction and Western blotting analyses were performed to detect the expression levels of CXCL12 mRNA and protein in primary mouse dermal fibroblasts treated with or without 5-FU. Transwell assays and fluorescein isothiocyanate (FITC)-phalloidin staining were used to observe cell migration and filamentous actin (F-actin) changes of melan-a murine melanocytes. RESULTS: Whole mount and cryosection X-gal staining showed that the cell numbers of LacZ-positive melanocytes were much higher in the margins of dorsal and tail skin wounds treated with 5-FU compared with the controls. Meanwhile, CXCL12 immunostaining was significantly increased in the dermal compartment of wounds treated with 5-FU (control vs. 5-FU, 22.47 ± 8.85 vs. 44.69 ± 5.97, P < 0.05). Moreover, 5-FU significantly upregulated the expression levels of CXCL12 mRNA (control vs. 5-FU, 1.00 ± 0.08 vs. 1.54 ± 0.06, P < 0.05) and protein (control vs. 5-FU, 1.00 ± 0.06 vs. 2.93 ± 0.10, P < 0.05) in cultured fibroblasts. Inhibition of the CXCL12/CXCR4 axis suppressed melanocyte migration in vitro using a CXCL12 small interfering RNA (siRNA) or a CXCR4 antagonist (AMD3100). CONCLUSION 5-FU possesses a pro-pigmentary activity through activation of the CXCL12/CXCR4 axis to drive the chemotactic migration of melanocytes.
Animals ; Cell Movement ; Cell Proliferation ; Chemokine CXCL12/genetics* ; Fibroblasts ; Fluorouracil/therapeutic use* ; Humans ; Mice ; RNA, Messenger ; Receptors, CXCR4