Objective To investigate the association of the expressions of p27 and proliferating cell nuclear antigen(PCNA)and nuclear-associated antigen(ki-67)proteins in synovial sarcoma,and the relationship with its prognosis.Methods The expression of p27,PCNA and ki-67 in 36 synovial sarcoma and 10 normal synovial tissue were immunohistochemically examined.The relationship between the expressions of p27,PCNA and ki-67 proteins in synovial sarcoma with its prognosis by clinical stage,tissue grades and prognosis was investigated.Results (1)In normal synovial tissue,the expressions of PCNA(0) and ki-67(0) were positive,p27 were positive100%(10/10).but the expressions of PCNA[86.1%(31/36)]and ki-67[69.4%(25/36)]in most synovial sarcoma were positive,p27 were negative[11.1%(4/36)],significant differences were found among these expressions(χ2=25.16,χ2=9.18,P＜0.05).(2) p27 positive-expression correlated significantly wiht thehistological grade of synovial sarcoma(χ2=9.19,P＜0.05)and prognosis(χ2=5.98,P＜0.05).The positive expressions of PCNA and ki-67 were positively related with the clinical stage(χ2=7.40,χ2=8.12,P＜0.05)and tissue grades(χ2=7.17,χ2=9.18,P＜0.05)and prognosis(χ2=13.03,χ2=10.66,P＜0.05)of synovial sarcina(3) There were negative correlations between p27 and PCNA(r=-0.63,P＜0.05) and between p27 and ki-67(r=-0.53,P＜0.05).But positive correlation between PCNA and ki-67(r=0.63,P＞0.05)was found.Conclusions Expressions of PCNA and ki-67 in most synovial sarcomas are positive,and that of p27 is negative,each plays an important role in the proliferation of synovial sarcoma.Combination of measuring of the p27,PCNA and ki-67 is valuble in predicting the prognosis.

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

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

Humans ; Membrane Glycoproteins ; immunology ; SARS Virus ; immunology ; Severe Acute Respiratory Syndrome ; prevention & control ; Spike Glycoprotein, Coronavirus ; Vaccines, DNA ; immunology ; Vaccines, Inactivated ; immunology ; Vaccines, Synthetic ; immunology ; Viral Envelope Proteins ; immunology ; Viral Vaccines ; immunology ; Virion ; immunology

Obej ctive Abnormal proliferation of T cells plays an important role in the development of autoimmune diseases. The article aimed to study the inhibitory effect of small molecule compound BD691 on T cell proliferation and its mechanism. Methods Human peripheral blood T-lymphocytes were isolated and purified by the immunomagnetic microbeads,then T cells were ac-tivated with anti-CD3/CD28 mAbs or alloantigen.The inhibitory effect of BD691 on activated T cell proliferation, the cytotoxic effect BD891 on resting T cells and the expression of activated T cells marker CD25 were measured by flow cytometry.Furthermore, ELISA was used to detect the secretion of cytokines associated with T cell differentiation. Results BD691 significantly inhibited the prolif-eration of T cells being stimulated by anti-CD3/CD28 mAb or alloantigen in a dose-dependent manner, and IC50 values are (8.5 ± 1.5)μmol/L and (7.2 ±1.3)μmol/L, respectively.However, BD691 had no obvious cytotoxic effects on resting T cells and periph-eral blood mononuclear cells, even at a high concentration ( up to 100μmol/L) .In T cells which were not activated by anti-CD3/CD28 mAb, the percentage of CD25+T cells is only 1.6%of the total cells, while the number increased to 68% after activating treatment.Mean-while, in T cells which were activated by 0, 3.3, 10, 30μmol/L BD691, no obvious change of CD25 expression were observed, while immunosuppressant FK506 (0.1μmol/L) significantly decreased the expression of CD25 +T cells (14.9%).In unactivated T cells, 95.6%cells were at G0/G1 phase, while after activation, the percentage of cells at G0/G1 phase reduced to 57.7%.In addition, BD691 inhibited the secretion of IFN-γ, IL-6 and IL-17 in activated T cells, but had no effects on the secretion of IL-2, IL-4 and IL-10. Co nclusion BD691 exerts no effects on T cell activation, but it inhibits T cell proliferation by inducing T cell cycling arrest at G0/G1 phase.Moreover, BD691 inhibits the secretion of key cytokines (such as IFN-γ, IL-6, IL-17) closely related to the differ-entiation of Th1 and Th17 cells.The results suggest that BD 691 is a potential lead compound to develop a new immunosuppressant for the inhibition of abnormal proliferation and differentiation of T cells.

In the present paper, the basic principles, the device and the analytical method of the hydrodynamic chromatography (HDC) were summarized, which is most widely used in hydrokinetic chromatography. The application of the hydrodynamic chromatography in the determination of the particle size and size distribution of the particulate drug delivery system was also reviewed. The method can determine the particle size of nano- and micron-scale particulate drug delivery systems rapidly. And this method also has the advantages of economic, convenient and no damage to the samples. In summary, there will be a good prospect for the application of HDC in the determination of particle size distribution features of particulate drug delivery systems.
Capsules ; chemistry ; Chromatography ; methods ; Drug Delivery Systems ; Hydrodynamics ; Liposomes ; chemistry ; Microspheres ; Nanoparticles ; chemistry ; Particle Size

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

OBJECTIVETo explore the analgesic mechanism of small knife needle for treating transverse process syndrome of the third vertebra (TPSTV) by observing peripheral and central changesof β-endorphin (β-EP) and enkephalin (ENK) contents.

METHODSTotally 30 Japanese white big-ear rabbits of clean grade were divided into 5 groups according to random digit table, i.e., the normal control group, the model group, the small knife needle group, the electroacupunture (EA) group, and the small knife needle plus EA group, 6 in each group. The TPSTV model was established by inserting a piece of gelatin sponge into the left transverse process of 3rd lumbar vertebrae. Rabbits in the small knife needlegroup were intervened by small knife needle. Those in the EA group were intervened by EA at bilateralWeizhong (BL40). Those in the small knife needle plus EA group were intervened by small knife needleand EA at bilateral Weizhong (BL40). Contents of β-EP and ENK in plasma, muscle, spinal cord, and hypothalamus were determined after sample collection at day 28 after modeling.

RESULTSCompared with the normal control group, contents of β-EP and ENK in plasma and muscle increased significantly, and contents of β-EP and ENK in spinal cord and hypothalamus decreased significantly in the model group (P < 0.05, P < 0.01). Contents of β-EP and ENK approximated normal levels in the three treatment groups after respective treatment. Compared with the model group, the content of β-EP in muscle decreased, and contents of β-EP and ENK in hypothalamus increased in the three treatment groups after respective treatment (P < 0.05). There were no significant difference among the three treatment groups (P > 0.05).

CONCLUSIONSSmall knife needle treatment and EA had benign regulation on peripheral and central β-EP and ENK in TPSTV rabbits. Small knife needle treatment showed better effect than that of EA.

Acupuncture Points ; Animals ; Electroacupuncture ; Enkephalins ; metabolism ; Hypothalamus ; metabolism ; Lumbar Vertebrae ; pathology ; Muscle, Skeletal ; metabolism ; Needles ; Rabbits ; Random Allocation ; Spinal Cord ; metabolism ; Spinal Diseases ; therapy ; beta-Endorphin ; metabolism

Background and purpose:It was reported that chemokine receptor CXCR4 and its ligand stromal cell-derived factor 1(SDF-1)were involved in the proliferation,differentiation,and metastasis of tumor.This study was designed to observe the expression of chemokine receptor CXCR4 in hypopharyngeal squamous cell carcinoma tissue and study the relationship between the expression of chemokine receptor CXCR4 and different clinicopathlogical characteristics,and further to explore the clinical significance.Methods:For the detection of the expression of chemokine receptor CXCR4,43 primary hypopharyngeal squamous cell carcinoma tissues,27 normal hypopharyngeal tissues,34 lymph node metastastatic lesions and 9 normal lymph node lesions were detected by immunohistochemical method using rabbit anti-human CXCR4 polyclonal antibody.Results:The positive expression rates of CXCR4 in 43 hypopharyngeal carcinoma tissues and normal tissues were 95.3% and 22.2%,respectively(P

Objective To explore acute toxicity of succimer on mice.Methods Twenty Kunming mice(10 males and 10 females) weighting approximately (21.2?2.3)g were acclimatized for 3 days prior to dosing,then were divided into control group and experiment group with 10 mice in each group according to body weight.Fasted for 12 hours,the mice in experiment group received intragastric administration of 160mg DMSA in deionized water in 24 hours,and the control group received the same volume of deionized water,and then they were observed for 7 days.Blood was collected into heparinized-tubes by removal of eyeball.All mice were sacrificed and brain,heart,liver and kidney were removed and washed with normal saline.The activity or amount of BUN,Scr,AST,ALT,SOD, GSH-PX and MDA were analyzed.Results (1)Given 160rag DMSA in 24 hours,gastrointestinal symptoms were main side effects.During the observation,experiment group lost weight due to the decrease of food-intake ,and some mice had slight hydroabdomen.(2)High dose of DMSA caused a significant inhibition of GSH-PX(P0.05).The hepatic cell was damaged accord- ing to the significant raise of MDA in liver(P0.05),which was related to acute toxicity on liver.Conclusion Succimer could inhibit the antioxidarrt systems and could do damage to liver and kidney.