OBJECTIVEBy using the RNAi method to inhibit Itk protein expression specificity, to observe lymphocytes proliferation and cytokines production, verify its function as a drug target.

METHODSDesigned siRNA aims at Itk sequence according to its sequence and solid structure, then electrotransfected into mouse spleen lymphocytes, We validated the decrease of Itk protein by Western-Blot, and detected the change of the cell proliferation by MTS and the change of inflammatory cytokines by ELISA.

RESULTSItk protein can be suppressed by Itk-siRNA, there were significantly reduced compared to its control group on cell proliferation as well as cytokine secretion such as IL-2, IL-4, IL-5, IFN-gamma. They all have statistical difference (P < 0.05).

CONCLUSIONItk has an important immunomodulatory effect in mouse spleen lymphocytes proliferation and secretion of inflammatory cytokines.This can supply an experimental basis to regard Itk as drug target for inflammation therapy.

Animals ; Cell Differentiation ; Cell Proliferation ; Cytokines ; genetics ; immunology ; Lymphocytes ; cytology ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Protein-Tyrosine Kinases ; genetics ; immunology ; Spleen ; cytology ; immunology

OBJECTIVETo observe the pathological changes and the expression of tumor necrosis factor alpha (TNF-alpha) and transforming growth factor beta (TGF-beta) in lung tissue of rats with radiation injury for exploring the mechanism of blood-activating and stasis-dissipating drugs in radiation injury prevention and treatment.

METHODSOne hundred and thirty SD female rats were randomly allocated into the simple irradiation group (n=60), the TCM herbs treatment group (n=60) and the blank control group (n=10). The right lung of all rats except those in the blank control group were irradiated by linear accelerator, 3 Gy each time, twice weekly, the maximum accumulated dose being 30 Gy. Ten rats in the two groups were randomly sacrificed at each of the 6 time points (1, 3, 5, 8, 12 and 26 weeks after repeated irradiation), their lung was harvested out, sliced and dyed with HE stain. The histological changes, levels of TNF-alpha and TGF-beta expression in the lung tissue were then observed by immunohistochemical technique.

RESULTSThe most serious acute radiation pneumonia was seen in the 5th week and pulmonary fibrosis was remarkable in the 26th week in the simple irradiation group, with the expressions of TNF-alpha and TGF-beta at different time phases enhanced significantly (P < 0.01). While in the TCM herbs treatment group, the pneumonia was milder, pulmonary fibrosis in late stage was not so obvious, and the expressions of TNF-alpha and TGF-beta significantly lower than those in the simple irradiation group (P < 0.01).

CONCLUSIONBlood-activating and stasis-dissipating drugs can inhibit expression of inflammation-inducing factors and fibrosis-inducing factors to lessen the inflammatory reaction of early radiation pneumonia, prolong the progression of radiation lung fibrosis, showing preventive and treating action on radiation lung injury.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Particle Accelerators ; Phytotherapy ; Radiation Injuries, Experimental ; drug therapy ; pathology ; prevention & control ; Radiation Pneumonitis ; drug therapy ; prevention & control ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; biosynthesis ; genetics ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

The water-soluble polypeptide drug oxytocin was encapsulated in liposomes by reverse-phase evaporation vesicle method to obtain oxytocin loaded liposomes (OT@LPs) which was further modified with cationic cell penetrating peptide—arginine octamer (R8) to get R8 modified oxytocin loaded liposomes (OT@LPs-R8) which showed enhanced mucoadhesive. The brain targeting efficiency was evaluated preliminarily after nasal administration. OT@LPs-R8 showed a round shape with a particle size distribution of 110.2 ± 7.3 nm, a surface potential as high as +18 mV, a drug loading (62.17 ± 1.88) %, an encapsulation rate (5.85 ± 0.72) %, and stood stable in nasal mucus. After nasal administration, it could significantly prolong the retention and enhance the distribution in the brain with no irritation to the nasal mucosa. The animal experiment in line with the regulations of the Department of Laboratory Animal Science of Fudan University on the ethics of animal experiments had been carried out after passing the review of the Animal Ethics Committee of Fudan University. The results showed nasal administration of OT@LPs-R8 could promote oxytocin directly into the brain from the nose which expected to become a new carrier for delivery of oxytocin to the brain.

AIM AND METHODSUsing Strain C57/BL of COX-2 deficient mice, the effect of Cyclooxygenase-2 (COX-2) on dopaminergic neurons in substantia nigra of Parkinson's disease (PD) mice induced by intraperitoneal injections of MPTP-HCl were investigated by immunocytochemistry(ICC) .

RESULTSWe found that the mortality in COX-2 heterozygous mice is much lower than that in wild type mice (P < 0.01) after injection of MPTP (30 mg/kg/day). The result of semiquantitative immunocytochemical staining showed that the number of tyrosine hydroxylase (TH) immunoreactive neurons in the substantia nigra pars compacta (SNc) declined more significantly in MPTP-treated wild type mice than that in COX-2 heterozygotes mice (P < 0.01).

CONCLUSIONCOX-2 may be related with lesion of dopaminergic neurons in the SNc of PD.

Animals ; Cyclooxygenase 2 ; deficiency ; Male ; Mice ; Mice, Inbred C57BL ; Neurons ; pathology ; Parkinson Disease ; pathology ; Substantia Nigra ; pathology

OBJECTIVETo observe the effects of cross electro-nape-acupuncture on reflex remodeling of airway protective reflex cough in patients with tracheotomy after cerebral hemorrhage.

METHODSWith the method of completely random design, according to treatment order, 60 patients who received tracheotomy after cerebral hemorrhage accompanied with cough reflex difficulty were randomly divided into a cross electro-nape-acupuncture group and an acupuncture group, 30 cases in each group. Both groups were treated with basic treatment, including anti-inflammation, eliminating phlegm, improving cerebral metabolism and so on. The acupuncture group was treated with acupuncture at Yifeng (TE 17), Fengchi (GB 20), Lianquan (CV 23), Baihui (GV 20), Touwei (ST 8), Dicang (ST 4) through Jiache (ST 6), Hegu (LI 4), Quchi (LI 11), and motor area on the affected side, and the needles were retained for 30 min. Based on the treatment of acupuncture group, the cross electro-nape-acupuncture group was additionally treated with cross electro-nape-acupuncture (continuous wave) for 30 min per treatment. The treatment was both given twice a day from Monday to Friday and once a day on Saturday and Sun day for 4 weeks. Tracheostomy cough reflex grading score (TCRGS) and clinical pulmonary infection score (CPIS) were observed before and after treatment in the two groups, and the clinical efficacy of two groups was evaluated.

RESULTSCompared before the treatment, TCRGS and CPIS were both reduced in two groups (both P < 0.01); after treatment, there were significant differences of TCRGS and CPIS between two groups (both P < 0.01), indicating cross electro-nape-acupuncture group was superior to acupuncture group. Regarding the effects of cough reflex remodeling, the cured and markedly effective rate was 96.7% (29/30) in the cross electro-nape-acupuncture group, which was significantly different from 55.2% (16/29) in the acupuncture group (P < 0.01).

CONCLUSIONCross electro-nape-acupuncture could effectively improve the remodeling of cough reflex and promote the recovery of lung infection in patients with tracheotomy after cerebral hemorrhage, leading to an increased quality of life.

Acupuncture Points ; Aged ; Cerebral Hemorrhage ; physiopathology ; surgery ; therapy ; Cough ; physiopathology ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Reflex ; Tracheotomy

To evaluate the effects of lentivirus-delivered short hairpin RNA (shRNA) on CVB3 infection in an animal model by RNA interference technique, we constructed a recombinant lentivirus expressing shRNA-3753 against the viral genome region 3753-3771, then transduced Lenti-sh3753 into mice infected with CVB3. We evaluated the antiviral ability of lenti-sh3753 by cytopathic effect (CPE), viral plaque assay and histological analysis of mice hearts. The results showed that Lenti-sh3753 exhibited a significant protective effect on cell viability and reduction of viral titers in supernatant of cell culture by specific inhibition on viral replication. Lenti-sh3753 also prolonged the mice survival and limited the viral production in mice hearts. These data proposed that Lenti-sh3753 can effectively inhibit CVB3 infection in a coxsackievirus-induced myocarditis model, suggesting its potential role in prevention and therapy of viral diseases.
Animals ; Coxsackievirus Infections ; drug therapy ; virology ; Down-Regulation ; Enterovirus B, Human ; genetics ; physiology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis ; drug therapy ; virology ; RNA Interference ; RNA, Small Interfering ; genetics ; therapeutic use ; RNA, Viral ; genetics ; Virus Replication

Objective To assess the feasibility of using PET molecular imaging to evaluate the therapeutic effects of traditional Chinese medicine FuZhiSan (FZS) on the model of aging Alzheimer's disease (AD) rats. Methods Twenty aged AD rats (Sparague-Dawley rats,male) were randomly divided into FZS treated group (n = 10) and control group (n = 10). Another 10 healthy adult rats were as blank controls. Morris water maze record system was used for cognitive function assessment. Before and after FZS treatment 18 F-fluorodeoxyglucose (FDG) and 11 C-2- [4'-(methylamino) phenyl] benzothiazol-6-ol ( PIB )PET imaging was undertaken. After post-treatment imaging procedures the brain tissues of all animals were taken for histochemical study,such as staining with HE,congo red,amyloid β (Aβ) immunofluorescence,5-bromo-2-deoxyuridine (BrdU) immunofluorescence and NeuN immunofluorescence. Paired t-test was performed with SPSS 13.0 software for the data analysis. Results The cognitive dysfunction of aging AD rats was improved after FZS treatment. The escape latency in FZS treated group was significantly shorter than that of control group ((32.5 ±10.8) s vs (102.6±8.8) s,t =15.7987,P=0. 0001). Diffuse neuronal loss and Aβ deposition were detected in the hippocampus and cortex in the aged AD rats. The imaging data showed that brain glucose metabolism was amended in FZS treated group while the abatement of amyloid deposition was not significant. Immunofluorescence results indicated that the neuronal proliferation was more remarkable in FZS treated group. Conclusions It may be feasible to use PET imaging as a method to evaluate the therapeutic effect in AD rats. FZS may ameliorate memory dysfunction of aged AD rats. Its mechanism may be partly contributed to the enhancement of the neuronal proliferation and survival.

The study was purposed to investigate the effect of alloreactive natural killer (alloNK) cells on immune reconstitution in murine haploidentical bone marrow transplantation (BMT). The murine model of haploidentical BMT was established by using (C57BL/6×BALB/c)BCF(1)(H-2(d/b)) mouse as the donor, and BALB/c (H-2(d)) mouse as the recipient. Recipient mice were divided into BMT group, non-allo-reactive NK (non-alloNK) cell group and alloNK cell group according to different transfusion. The effect of adding alloNK cells to transfusion was assessed by thymus pathology, the proportion of spleen NK cells, the spleen cell proliferation, the IFN-γ and IL-4 concentrations product at 24 and 48 h of recipient spleen cell culture supernatant at 2 months after BMT. The results showed that there were no obvious difference in thymus tissue among 3 groups under the optical microscope. The proportion of recipient spleen NK cells in non-alloNK group was significantly lower than that in BMT group (P < 0.05). There was no significant difference in proliferation of the recipient spleen cells among 3 groups at 2 months after BMT. The IFN-γ concentration product at 24 and 48 h of recipient spleen cell culture supernatant in alloNK group was significantly lower than that in other 2 groups at 2 months after BMT (P < 0.05). The IL-4 concentration in each group was not significantly different (P > 0.05). It is concluded that alloNK cells do not damage the thymus structure and may induce Th2 immune response in murine haploidentical BMT.
Animals ; Bone Marrow Transplantation ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Killer Cells, Natural ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Spleen ; cytology ; Thymus Gland ; pathology ; Transplantation, Homologous

OBJECTIVETo systematically evaluate the effect and safety of Xuezhikang Capsule (XZKC) for adjuvant treatment for coronary heart disease (CHD) patients accompanied with or without dyslipidemia.

METHODSChina National Knowledge Infrastructure (CNKI) Database, Chongqing VIP Database (VIP), Wanfang Data base, Cochrane Library, and Medline (PubMed) were retrieved with the deadline of August 30, 2013. Randomized controlled trials (RCT) of XZKC in treating CHD patients with or without dyslipidemia were all included. Assessment of bias risk for included studies was conducted according to the Cochrane Handbook for Systematic Reviews of Intervention (Version 5.0.2): Criteria for judging risk of bias in the "risk of bias" assessment tool. Review Management (5.1.0) was employed for data statistics. If there was no significant heterogeneity, results from the random-effect model were presented. If the heterogeneity was not substantial, a meta-analysis was not performed and a narrative and qualitative summary was performed instead.

RESULTSA total of 28 RCTs (6,949 patients) were included after screening results. The methodological quality of included trial was generally lower. Results of Metaanalysis showed that XZKC was beneficial for CHD patients in decreasing cardiovascular events: when compared with the basic treatment group, the relative risk (RR) was 0.53 and 95% confidence interval (CI) was [0.35, 0.81]; when compared with the placebo + basic treatment group, RR was 0.52 and 95% CI was [0.42, 0.65]; when compared with the basic treatment group, RR for improving symptoms of angina was 1.20 and 95% CI was [1. 12, 1.30]; when compared with the basic treatment group, RR for improving abnormal ECG was 1.38 and 95% CI was [1.21, 1.57]. Thirteen studies showed that XZKC + basic treatment was obviously superior in lowering total cholesterol (TC) to that of the basic treatment group. Three studies showed that XZKC + basic treatment was obviously superior in lowering total cholesterol (TC) to that of the placebo + basic treatment group. Thirteen studies showed that XZKC + basic treatment was obviously superior in lowering low density lipoprotein cholesterol (LDL-C) to that of the basic treatment group. Three studies showed that XZKC + basic treatment was obviously superior in lowering LDL-C to that of the placebo + basic treatment group. A total of 18 studies describing adverse reactions (ADs) involved 61 ADs in the XZKC + basic treatment group. All suffered from mild symptoms or were improved after treatment. No severe ADs occurred.

CONCLUSIONTreatment of CHD by XZKC might lower the occurrence of cardiovascular events in CHD patients accompanied with or without dyslipidemia, relieve clinical symptoms, improve ECG, lower blood lipid levels, and with less adverse reactions.

Angina Pectoris ; Cardiovascular Diseases ; Combined Modality Therapy ; Confidence Intervals ; Coronary Disease ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Randomized Controlled Trials as Topic

Objective To establish an automatic synthesis method for 11C-carfentanil (CFN) as an novel opiate receptor radioligand and study its biodistribution in rats. Methods 11C-Triflate-CH3 was bubbled into 0.5 mg precursor desmethyl-CFN (which was dissolved in 0.15 ml DMSO) to generate 11C-CFN in a V-tube at room temperature. Sep-Pak C2 column was used for purification of 11C-CFN, which was eluted by 3ml binary system aqueous solution, 10 ml water thrice, and then I ml ethanol. The biodistribution (% ID/g) of 11C-CFN in SD rats was studied. SPSS 13.0 was used for statistical analysis. Non-normal distribution data were analyzed using nonparametric test. Results The synthesis time for 11C-CFN was 20 min (end of bombardment, EOB). The synthesis yield was (35.5 ± 2.2) % on average (n = 12, uncorrected)with the radiochemical purity over 98%. Biodistribution study in rats showed that the tracer had a high brain uptake, rapid blood clearance, and a metabolic pathway via liver and kidney. The highest tracer uptake was in thalamus (4.26 ± 0.89) % ID/g and striatum (4.05 ± 1.08) % ID/g at 5 min after injection, followed by cerebral cortex (2.63±0.89) %ID/g, pons (2.26 ±0.57) % ID/g, hippocampus (2. 17 ±0.55) %ID/g and cerebellum (2. 15 ±0.39) %ID/g. Conclusions The automatic synthesis of 11C-CFN is fast and reliable, and this radioligand can be used for opiate receptor imaging.