Nature is abundant in protein scaffolds.By selecting suitable protein scaffold,display and screening methods,the rational and constrained random peptide library(RPL)can be constructed.Compared with the non-constrained RPL,it offered more opportunities for obtaining novel protein structures and more higher affinity ligands against the target molecules.At present,the protein scaffold constrained RPLs have been shown great potential in applications such as target selection,basic research,clinical diagnosis,medical therapy and so on.It is systematically introduced the structure bases,classification and construction of constrained RPL based on scaffolds,as well the recent great advances of application in selection against target molecules with S-S constrained scaffolds,antibodies,Zinc finger protein,Z domain,FN3 domain as important examples.

Objectives To investigate the prevalence of animal plague in Horqin Right Front County and provide scientific bases for prevention and treatment of the disease.Methods Plague surveillance was carried out according to the requirements of National Plague Monitoring Programme at 3 monitoring points in Horqin Right Front County in 2012.Squirrel density,the number of small rats,rat fleas,pathogenic and serological surveillance of Yersinia pestis were included in the study.Results A total of 356 dauricus were captured,the average density was 0.88 rats/hm2; female and male ratio was 1 ∶ 1.31 and the female embryo rate was 96.46％.The captured 13 small nocturnal mice belonged to 2 species,which included 10 cricetulus barabensis and 3 cricetulus longicaudatus,and the capture rate of small nocturnal mouse was 4.33％ (13/300).A total of 98 rat fleas were collected from 25 rats; rat flea carrying rate was 100％(25/25) and flea index was 3.92.Eighty rat burrows were detected,and flea index was 0.188.Pathogen test results were negative.Indirect hemagglutination test was used to examine 356 rat sera and 1 serum was positive.Conclusions Detection of positive rat sera has proved the existence of plague epidemic in Horqin Right Front County.Therefore,the work on plague prevention and control has a long way to go.

Objective To study the effect of different concentration of 1,25-dihydroxyvitamin D3[1,25(OH)2D3] on cell proliferation,differentiation and the expression of vitamin D receptor (VDR) in mouse MC3T3E1 osteoblast.Methods Osteoblast were cultured in medium with different concentrations of 1,25(OH)2D3.Incubated for 48 h,cell proliferation of osteoblast were examined by MTT reduction assay (mono-nuclear cell direc cytotoxicity assay),the osteocalcin (OC) levels in cell medium were detected by ELISA,and the expression of VDR mRNA and protein were examined by using SYBR Green real-time PCR and Western blot,respectively.Results 1.After incubation with 1,25(OH)2D3 for 48 h,the number of MC3T3E1 osteoblast was significantly less than that in control group(P0.05).3.SYBR Green real-time PCR and Western blot results showed that the expression of VDR mRNA as well as VDR protein of osteoblast in 10-8,10-9 mol/L experimental groups were significantly higher than those in control group (Pa0.05).Conclusions Cell proliferation of mouse osteoblast can be inhibited,while the cell differentiation was promoted by 1,25(OH)2D3.1,25(OH)2D3 up-regulated the expression of VDR in mouse osteoblast,which suggested that the VDR signal pathway may play some role in proliferation and differentiation of osteoblast.

Our previous results have demonstrated that insulin reduces myocardial ischemia/reperfusion (MI/R) injury and increases the postischemic myocardial functions via activating the cellular survival signaling, i.e., phosphatidylinositol 3-kinase (PI3-K)-Akt-endothelial nitric oxide synthase (eNOS)-nitric oxide (NO) cascade. However, it remains largely controversial whether c-Jun NH2-terminal kinase (JNK) is involved in the effects of insulin on MI/R injury. Therefore, the aims of the present study were to investigate the role of JNK, especially the cross-talk between JNK and previously expatiated Akt signaling, in the protective effect of insulin on I/R myocardium. Isolated hearts from adult Sprague-Dawley rats were subjected to 30 min of regional ischemia and followed by 2 or 4 h of reperfusion (n=6). The hearts were pretreated with PI3-K inhibitor LY294002, or phosphorylated-JNK inhibitor SP600125, respectively, then perfused retrogradely with insulin, and the mechanical functions of hearts, including the heart rate (HR), left ventricular developed pressure (LVDP) and instantaneous first derivation of left ventricular pressure (+/-LVdp/dt(max)) were measured. At the end of reperfusion, the infarct size (IS) and apoptotic index (AI) were examined. MI/R caused significant cardiac dysfunction and myocardial apoptosis (strong TUNEL-positive staining). Compared with the control group, insulin treatment in MI/R rats exerted protective effects as evidenced by reduced myocardial IS [(28.9 +/- 2.0)% vs (45.0 +/- 4.0) %, n=6, P<0.01], inhibited cardiomyocyte apoptosis [decreased AI: (16.0 +/- 0.7) % vs (27.6 +/- 1.3) %, n=6, P<0.01] and improved recovery of cardiac systolic/diastolic function (including LVDP and +/-LVdp/dt(max)) at the end of reperfusion. Moreover, insulin resulted in 1.7-fold and 1.5-fold increases in Akt and JNK phosphorylation in I/R myocardium, respectively (n=6, P<0.05). Inhibition of Akt activation with LY294002 abolished, and inhibition of JNK activation with SP600125 enhanced the cardioprotection by insulin, respectively. And the abolishment by LY294002 could be partly converted by SP600125 pretreatment. In addition, SP600125 also decreased the Akt phosphorylation (n=6, P<0.05). These results demonstrate that insulin simultaneously activates both Akt and JNK, and the latter further increases the phosphorylation of Akt which attenuates MI/R injury and improves heart function; this cross-talk between Akt and JNK in the insulin signaling is involved in insulin-induced cardioprotective effect.
Animals ; Apoptosis ; Heart ; Insulin ; metabolism ; JNK Mitogen-Activated Protein Kinases ; MAP Kinase Signaling System ; Myocardial Infarction ; Myocardial Ischemia ; Myocardial Reperfusion Injury ; Myocardium ; Myocytes, Cardiac ; Nitric Oxide Synthase Type III ; Phosphatidylinositol 3-Kinase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; Signal Transduction

To study the anti-tumor metastatic constituents in Rhodiola wallichiana (HK) S H Fu var Cholaensis (Praeg) S H Fu, chemical constituents were isolated and purified by repeated column chromatography (silica gel, Toyopearl HW-40C and preparative HPLC). Their structures were elucidated on the basis of spectral data analysis. The anti-tumor metastasis assay was applied to evaluate the activities of the isolated compounds. Ten compounds (1-10) were isolated and their structures were identified by comparison of their spectral data with literature as follows: syringic acid (1), salidroside (2), tyrosol (3), scaphopetalone (4), berchemol (5), 2,6-dimethoxyacetophenone (6), rhobupcyanoside A (7), miyaginin (8), chavicol-4-O-β-D-apiofuranosyl-(1 --> 6)-O-β-D-glucopyranoside (9), eugenyol-O-β-D-apiofuranosyl-(1 --> 6)-O-β-D-glucopyranoside (10). Compounds 4-6 and 8-10, were isolated from this genus for the first time, while compound 7 was isolated from this plant for the first time. Compounds 2, 6-8 showed positive anti-tumor metastatic activities, and compounds 2 and 8 showed significant anti-tumor metastatic activities.
Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Humans ; Neoplasm Metastasis ; prevention & control ; Rhodiola ; chemistry

OBJECTIVETo observe the clinical effects of Prunellae Oral Liquid (POL) on the size of thyroid with different states of thyroid function in patients of goiter.

METHODSFifty-six patients with hyperthyroidism, 24 with hypothyroidism and 18 with simple goiter were randomized into two groups. Group A treated by classical therapy, Group B by POL based on classical therapy. Size of thyroid of patients before and after treatment was measured by color ultrasonic Doppler and compared with the data obtained from 20 healthy subjects as control.

RESULTSSize of thyroid in all patients before treatment were obviously larger than that of normal control (P<0.01), but it reduced after treatment in both groups, especially in group B, in which it differed insignificantly to that in the normal control (P > 0.05).

CONCLUSIONCombined treatment of Chinese medicine (using POL) and Western medicine is superior to Western medicine alone in treating goiter with different states of thyroid function.

Adult ; Antithyroid Agents ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Goiter ; drug therapy ; physiopathology ; Humans ; Hyperthyroidism ; drug therapy ; physiopathology ; Hypothyroidism ; drug therapy ; physiopathology ; Male ; Middle Aged ; Phytotherapy ; Plant Extracts ; therapeutic use ; Prunella ; chemistry ; Thyroid Gland ; drug effects ; physiopathology ; Treatment Outcome

To explore the expression potential of heterogeneous genes using the backbone of infectious bronchitis virus (IBV) Beaudette strain, the ectodomain region of the Spike gene (1,302 bp) of IBV H120 strain was amplified by RT-PCR and replaced into the corresponding location of the IBV Beaudette strain full-length cDNA. This recombinant was designated as BeauR-H120(S1). BeauR-H120(S1) was directly used as the DNA template for the transcription of viral genomic RNA in vitro. Then, the transcription product was transfected into Vero cells by electroporation. At 48 h post-transfection, the transfected Vero cells were harvested, and passaging continued. A syncytium was not observed until the recombinant virus had passed through four passages. The presence of rBeau-H120(S1) was verified by the detection of the replaced ectodomain region of the H120 Spike gene using RT-PCR. Western blot analysis of rBeau-H120 (S1)-infected Vero cell lysates demonstrated that the nucleocapsid (N) protein was expressed, which implied that rBeau-H120(S1) could propagate in Vero cells. The TCIDs0 and EIDs0 data demonstrated that the titer levels of rBeau-H120(S1) reached 10(590+/-0.22)TCID50/mL and 10(6.13+/-0.23)EID50/mL in Vero cells and 9-day-old SPF chicken embryos, respectively. Protection studies showed that the percentage of antibody-positive chickens, which were vaccinated with rBeau-H120(S1) at 7 days after hatching, rose to 90% at 21 days post-inoculation. Inoculation provided an 85% rate of immune protection against a challenge of the virulent IBV M41 strain (103EID50/chicken). This recombinant virus constructed using reverse genetic techniques could be further developed as a novel genetic engineering vaccine against infectious bronchitis.
Animals ; Cercopithecus aethiops ; Chick Embryo ; Chickens ; Coronavirus Infections ; veterinary ; virology ; Infectious bronchitis virus ; chemistry ; genetics ; growth & development ; metabolism ; Poultry Diseases ; virology ; Protein Structure, Tertiary ; Spike Glycoprotein, Coronavirus ; chemistry ; genetics ; metabolism ; Transfection ; Vero Cells

The genus Xestospongia is one of the most widespread genera of sponges, containing abundant secondary metatolites with novel structures and potent bioactivities. The main structure types of secondary metatolites found in this genus are alkaloids, quinines, terpens, steroids, lipids, polyketones, etc. These metatolites exhibit a variety of bioactivities, such as cytotoxic, antibacterial and antiviral activities. This paper reviews the progress in the chemistry and pharmacological activities of the second metabolities from sponges of Xestospongia, especially for recent five years, with the aim for further research.
Animals ; Secondary Metabolism ; Xestospongia ; chemistry

AIMTo investigate the effect of 8-opioid receptors in the cardioprotection elicited by ischemic postconditioning and the underlying mechanism.

METHODSThe isolated perfused hearts of male Sprague-Dawley rats were subjected to 30 min of global ischemia followed by 120 min of reperfusion. Formazan content of myocardium was measured spectrophotometrically, and the activity of lactate dehydrogenase (LDH) in the coronary effluent was measured. In isolated ventricular myocytes hypoxic postconditioning was achieved by 3 cycles of 5 min reoxygenation/5 min hypoxia starting at the beginning of reoxygenation, and cell viability was measured.

RESULTSIn the Langendorff perfused rat heart model, ischemic postconditioning (6 cycles of 10 s reperfusion/10 s global ischemia starting at the beginning of reperfusion) increased formazan content, reduced LDH release, improved the recovery of the left ventricular developed pressure, maximal rise/fall rate of left ventricular pressure and rate pressure product (left ventricular developed pressure multiplied by heart rate), attenuated the decrease of coronary flow during reperfusion and increased the isolated cell viability. Pretreatment with naltrindole, an antagonist of delta-opioid receptors and calcium-activated potassium channel (KCa) blocker paxilline attenuated the effect of ischemic/hypoxic postconditioning.

CONCLUSIONThe findings indicate that ischemic postconditioning protects myocardium against ischemia/reperfusion injury via activating delta-opioid receptors and opening KCa.

Animals ; Cell Survival ; In Vitro Techniques ; Ischemic Postconditioning ; Male ; Myocardium ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Potassium Channels, Calcium-Activated ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, delta ; antagonists & inhibitors

This study is to investigate the effect of ZL-004 on normal mouse and mice with leukopenia induced by chemotherapeutic agents. 5-Fluorouracil were administered intraperitoneally to mice to develop leucopenia, and the mice were treated with ZL-004. The number of peripheral leukocytes and the percentage of granulocyte in total WBC were examined. The results are that ZL-004 markedly raise peripheral blood leukocytes in the normal mice and the mice model of leukopenia. So, ZL-004 could protect mice against 5-fluorouracil damage and raise peripheral blood leukocyte. Features of bone marrow smears is myeloproliferative hyperactivity in the mice, particularly the matured granulocytic series were observed. The mechanism of ZL-004 is to act on the mouse bone marrow causing proliferation and differentiation.
Animals ; Antineoplastic Agents ; pharmacology ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Female ; Fluorouracil ; Granulocytes ; cytology ; drug effects ; Leukocyte Count ; Leukocytes ; cytology ; drug effects ; Leukopenia ; chemically induced ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Structure ; Pyrrolidinones ; chemistry ; pharmacology ; Random Allocation