The baculovirus expression system has been used extensively for the expression of recombinant proteins in insect cells.Recently,recombinant baculoviruses containing mammalian cell-active promoter element have been used to transduce a broad spectrum of primary and established mammalian cells,including non-hepatic cells.Recombinant baculoviruses have been used successfully for transient or stable gene delivery in mammalian cells in vitro,while the efficiency of delivering gene in vivo is inhibited obviously by complements,but efforts have been made to overcome the problems,for instance,VSV-G-pseudotyped baculoviruses display complement resistance.The mechanism of the transduction is still not clearly understood,though many researches have been done.The baculovirus is able to replicate only in insect cells,but it is incapable of initiating replication cycle in mammalian cells,which guarantees high biosafety of this gene delivery system.In addition,this system is easily manipulated and able to carry large inserts.These attributes will undoubtedly lead to the increased application and continued development of this system for highly effective gene delivery into mammalian cells.

AIMTo increase the cognition of cerebellar functions and the knowledge of neuroimmunology, the effect of cerebellar interposed nuclei (IN), one of three deep nuclei in cerebellum, on lymphocyte function was investigated.

METHODSKainic acid (KA) was microinjected into bilateral IN for lesions of neuronal bodies in the IN. Control rats was microinjected with saline into their IN. On days 8, 16 and 32 following the IN lesions, the lymphocyte number in the peripheral blood was measured by blood corpuscle counter. Meanwhile, lymphocyte proliferation induced by concanavalin A (Con A), cytotoxicity of natural killer (NK) cells against YAC-1 cells, and anti-SRBC IgM antibody in the serum were examined respectively by methyl-thiazole-tetrazolium (MTT) assay, flow cytometry and ELISA assay.

RESULTSThe lymphocyte number in the peripheral blood was significantly reduced on days 8, 16 and 32 following the effective lesions of the bilateral IN in comparison with that of control. The Con A-induced lymphocyte proliferation, the NK cell cytotoxicity to YAC-1 cells, and the titer of anti-SRBC IgM antibody in the serum, were all significantly attenuated on days 8, 16 and 32 following the effective lesions of the bilateral IN in comparison with those of control. There were not remarkable differences between the days 8, 16 and 32 in the decreased lymphocyte number and functions induced by the lesions of the bilateral IN.

CONCLUSIONEffective lesions of the cerebellar bilateral IN of rats cause an inhibition in lymphocyte number and functions of T, B and NK cells, strongly showing that the cerebellar IN can modulate lymphocyte functions.

Animals ; Cerebellar Nuclei ; immunology ; physiology ; Cerebellum ; immunology ; physiology ; Female ; Kainic Acid ; Killer Cells, Natural ; immunology ; Lymphocyte Count ; Lymphocytes ; immunology ; Male ; Microinjections ; Neuroimmunomodulation ; immunology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective To investigate the association between single nucleotide polymorphisms (SNPs)in cytokine IL-6, IL- 10 genes and HBV-related hepatocellular carcinoma(HCC). Methods A hospital-based case-control study was conducted in 381 cases with HBV-related HCC, 340 HBsAg carriers and 359 non-tumor controls. Genotypes of-572 site of IL-6 gene and-819, -592 sites of IL-10 gene were determined by real-time polymorphism chain reaction. Unconditional logistic regression was used to estimate the odds ratios(ORs)and 95 confidence intervals(C/s). Results For the G/C alleles of -572 loci on IL-6 gene, there were significant differences between the three groups(P＜0.05). Compared with CC genotype, GG genotype increased the risk of HBV infection (OR=2.171,95% Ch 1.068-4.415), but did not seem to be associated with HCC. For the alleles of-819 and -592 site of IL-10 gene, there were significant differences between the three groups(P＜0.05). Compared with CC genotype, TT genotype increased the risks of both HCC(OR=2.791,95%CI:1.326-5.874), and HCC in HBsAg carriers(0R=3.522,95%CI: 1.707-7.266). When compared with CC genotype on -592 site, the AA genotype reduced the risk of both HCC(OR=0.389, 95% CI:0.173-0.875), and HCC in HBsAg carriers(OR=0.336, 95% CI: 0.154-0.734). Conclusion The SNPs in -572 site of IL-6 gone might be associated with the risk of HBV infection. The SNPs in -819 site of IL-10 gene increased the risk of HCC, but -592 site of IL-10 gene decreased the risk of HCC.

OBJECTIVEParkinson's disease (PD), a neurodegenerative disorder, has been reported to be associated with brain neuroinflammation in its pathogenesis. Herein, changes in peripheral immune system were determined to better understand PD pathogenesis and provide possible target for treatment of PD through improvement of immune disorder.

METHODS1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was intraperitoneally injected into mice to prepare PD model. Expression levels of pro-inflammatory and anti-inflammatory cytokines and transcription factors of CD4+ T lymphocyte subsets in spleen and mesenteric lymph nodes and concentrations of the cytokines in serum were examined on day 7 after MPTP injection. Percentages of CD4+ T lymphocyte subsets were measured by flow cytometry.

RESULTSMPTP induced PD-like changes such as motor and behavioral deficits and nigrostriatal impairment. Expression levels of the pro-inflammatory cytokines including interferon (IFN)-γ, interleukin (IL)-2, IL-17 and IL-22, in spleen and mesenteric lymph nodes were upregulated and their concentrations in serum were elevated in PD progression. But, the concentrations of the anti-inflammatory cytokines including IL-4, IL-10 and transforming growth factor (TGF)-β were not altered in the two lymphoid tissues or serum of PD mice. In addition, expression of T-box in T cells (T-bet), the specific transcription factor of helper T (Th) 1 cells, was downregulated, but expression of transcription factor forkhead box p3 (Foxp3), the transcription factor of regulatory T (Treg) cells, was upregulated. In support of the results, the numbers of IFN-γ-producing CD4+ cells (Th1 cells) were reduced but CD4+CD25+ cells (Treg cells) were elevated in both the lymphoid tissues of PD mice.

CONCLUSIONPD has a dysfunction of peripheral immune system. It manifests enhancement of proinflammatory response and CD4+ T cell differentiation bias towards Treg cells away from Th1 cells.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine ; Animals ; CD4-Positive T-Lymphocytes ; pathology ; Cell Differentiation ; Cytokines ; blood ; Disease Models, Animal ; Flow Cytometry ; Forkhead Transcription Factors ; metabolism ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-17 ; blood ; Interleukin-2 ; blood ; Interleukin-4 ; blood ; Interleukins ; blood ; Lymph Nodes ; cytology ; Lymphocyte Activation ; Mice ; Parkinson Disease ; immunology ; physiopathology ; Spleen ; cytology ; T-Box Domain Proteins ; metabolism ; T-Lymphocytes, Regulatory ; Th1 Cells ; Transforming Growth Factor beta ; blood

OBJECTIVEWe used an animal model of collagen-induced arthritis (CIA) to study changes and roles of tyrosine hydroxylase (TH) expressed by CD4+ T cell subsets, and then explore the relationship between CD4+ T cell subset-derived catecholamines and inflammatory responses in CIA.

METHODSThirty-six male DBA/1 mice were randomly divided into control group, CIA model group (day 35) and CIA model group (day 55) (n = 12). CIA model was induced by type II collagen (CII) in DBA/1 mice. On the 35th and 55th day following primary immunization, the joints of the mice were observed for clinical score of swelling and the level of anti-CII IgG antibody in serum was examined. Expression of specific transcription factors and cytokines of Th1, Th17, Th2 and Treg cells and TH in mesenteric lymph nodes was measured by means of Western blot. The changes of TH expressed by CD4+ T cell subsets in mesenteric lymph nodes were determined by flow cytometry.

RESULTSClinical score and anti-CII antibody level increased in CIA compared with that in intact mice. Specific transcription factors and cytokines expressed by Th1 and Th17 cells were upregulated and cytokines expressed by Th2 and Treg cells were downregulated in mesenteric lymph nodes in CIA mice. Expression of TH was upregulated and the increased expression of TH in CD4+ T cells was attributed to Th1 and Th17 cells in mesenteric lymph nodes of CIA.

CONCLUSIONThe increase in catecholamines from CD4+ T cell subsets in mesenteric lymph nodes of CIA may be related to inflammatory alleviation in CIA progression.

Animals ; Arthritis, Experimental ; immunology ; CD4-Positive T-Lymphocytes ; enzymology ; Collagen Type II ; adverse effects ; Cytokines ; metabolism ; Disease Models, Animal ; Lymph Nodes ; immunology ; Male ; Mice ; Mice, Inbred DBA ; T-Lymphocyte Subsets ; immunology ; Transcription Factors ; metabolism ; Tyrosine 3-Monooxygenase ; metabolism

Professor LI Zhi-dao's acupuncture technique at muscle belly and its clinical experiences for the treatment of arthralgia are introduced. From aspects of TCM, anatomical physiology and sports biomechanics, the theory basis of this technique is explained, while its method is elaborated in details from acupoint selection and manipulation combined with classic medical cases. The acupuncture technique at muscle belly is applied at the muscles which already have pathological changes, avoiding re-stimulation on the pain area. It provides new thinking methods and references for acupuncture to treat arthralgia.
Acupuncture Points ; Acupuncture Therapy ; methods ; Adult ; Aged ; Arthralgia ; therapy ; Female ; Humans ; Male ; Middle Aged

To further explore the result of bilingual teaching in pediatrics,we randomly chose 200 undergraduates of 4 class and released students'questionnaires about bilingual teaching with teaching content before and after class to assess students'understanding of bilingual teaching and analysed appraisal result.We found no significant difference of student score between students accepting bilingual teaching and not accepting the bilingual teaching,but there was difference for English tests and expression level.So we think that students can fully accept the bilingual teaching of pediatrics under the premise with selecting appropriate teaching methods and means.

Adolescent ; Child ; Cord Blood Stem Cell Transplantation ; Female ; Flow Cytometry ; Humans ; Leukocyte Count ; Male ; Siblings ; Transplantation, Homologous ; immunology ; Treatment Outcome

AIMTo explore IL-6 neuroprotection against glutamate-induced neurotoxicity and primary mechanisms involved in this neuroprotection.

METHODSThe cerebellar granule neurons from postnatal 8-day infant rats were chronically exposed to IL-6 for 8 days, and then glutamate stimulated the cultured cerebellar granule neurons for 15 min. Methyl-thiazole-tetrazolium (MTT) assay and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method were used to observe the changes of neuronal vitality and apoptosis, respectively. Laser scanning confocal microscope (LSCM) and reverse transcription-polymerase chain reaction (RT-PCR) were respectively employed to measure dynamic changes of intracellular Ca2+ levels and expression of gp130 mRNA, a 130-kDa intracellular IL-6 signal-transduction protein, in the neurons.

RESULTSThe chronic IL-6 (2.5, 5 and 10 ng/ml) pretreatment of the cultured cerebellar granule neurons remarkably improved the decreased neuronal vitality by glutamate in a concentration-dependent manner. The neuronal apoptosis induced by glutamate was significantly attenuated by the chronic IL-6 pretreatment. The intracellular Ca2+ overload evoked by glutamate was also inhibited by the chronic IL-6 pretreatment. The expression of gp130 mRNA was dramatically lower in the IL-6-pretreated cerebellar granule neurons than in the IL-6-untreated neurons.

CONCLUSIONIL-6 can protect neurons against glutamate-induced exciting neurotoxicity. The mechanism of IL-6 neuroprotection may be closely related to the suppression of glutamate-induced intracellular Ca2+ overload and mediated by gp130 intracellular signal transduction pathways.

Animals ; Cells, Cultured ; Cerebellum ; cytology ; drug effects ; metabolism ; Glutamic Acid ; toxicity ; Interleukin-6 ; pharmacology ; Neurons ; drug effects ; metabolism ; Neuroprotective Agents ; pharmacology ; Neurotoxicity Syndromes ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the efficacy and safety of zhongfu hypotension capsule (ZHC) in treating hypertension of yin-deficiency with sthenic-yang syndrome type.

METHODSAdopting the stratified, block randomized, double-blinded, double-dummy, positive parallel controlled, multi-centered clinical trial method, the tested group was treated by orally taken 3 capsules of ZHC (0.5 g/capsule) twice a day, and the control group was treated by orally taken Lotensin Tablet 1 tablet (10 mg/tab.) once a day. And all received the adiaphorous drug with the dosage-form mimic to that used in the tested group. The therapeutic course was 4 weeks for both groups.

RESULTSThe markedly effective rate and the total effective rate in reducing blood pressure was 58.65% and 79.81% respectively in the tested group, and 60.51% and 78.34% respectively in the control group; while the markedly effective rate and the total effective rate for alleviating TCM syndrome was 21.15% and 78.85% in the tested group, and 25.48% and 86.62% in the control group respectively. Comparisons between the two groups showed insignificant difference (P > 0.05).

CONCLUSIONZHC has good efficacy and is safety in treating hypertension.

Adult ; Aged ; Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Benzazepines ; therapeutic use ; Blood Pressure ; drug effects ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypertension ; drug therapy ; physiopathology ; Male ; Middle Aged ; Phytotherapy ; Syndrome ; Tablets ; Yin Deficiency ; drug therapy ; pathology