Objective To explore the application of particle-gun technique in transfecting dendritic cells(DCs),and investigate the expression of DNA vaccine particle IgHV1-GM-CSF in DCs induced by particle-gun.Methods Monocytes were isolated from donor peripheral blood collected by Ficoll density gradient method and adherent culture,and then differentiated into immature DCs(imDCs) by rhGM-CSF and rhlL-4 induction.The plasmids for transfection were IgHV1-GM-CSF/pcDNA3.0 and pEGFP,which were prepared with endofree plasmid purification kit.The DCs were transfected with plasmid pEGFP by particle-gun at different transfection conditions,the green fluorescence positive cell and the total cell numbers were counted respectively under fluorescence microscope,and the transfection efficiency was calculated;the viable cell count was performed after trypan blue staining;the transfection efficiencies of particle-gun,liposome-mediated transfection and electroporation method were compared.The plasmids IgHV1-GM-CSF/pcDNA3.0 and pEGFP were cotransfected into imDCs by particle-gun under the optimum conditions,and then DNA was extracted,the expression of IgHV1-GM-CSF was determined by RT-PCR,and of GM-CSF by ELISA.Results The optimum conditions of particle-gun transfection in imDCs were: gas pressure at 120psi,PVP concentration in 0.02mg/ml,microcarrier loading in 0.5mg gold/shot,and DNA loading ratio in 1.5?g plasmid/mg gold.The transfection efficiencies of particle-gun and electroporation were 10.5%?2.4%(n=3) and 45.2%?5.6%(n=3) respectively,while that of liposome-mediated transfection was very low,and significant difference existed among the three methods(P

Objective To investigate the correlation between ~(18)F-fluorodeoxyglucose (FDG) uptake and hypoxia inducible factor1α (HIF-1α) level,microvessel density (MVD) in human gliomas.Methods ~(18)F-FDG PET scan was performed preoperatively in 41 patients with gliomas (including 23 highgrade and 18 low-grade tumors).The ratios of maximum standardized uptake value(SUV_(max))between tumor (T)and contralateral white matter (WM) were calculated (T/WM).Immunohistochemical stain methods were used to evaluate the level of HIF-1α and measure the MVD in tumors.Correlation analysis between SUV_(max) of T/WM and HIF-1α level,MVD wag performed.The t-test,one-way ANOVA test,Spearman rank correlation and Wilcoxon signed-rank test were calculated using SPSS 11.5 software.Results (1)The SUV_(max) of T/WM,HIF-1α level and MVD in high-grade and low-grade tumors groups were 3.39±1.43,95.7% and 44.13±16.1 vs 1.46±0.55.55.6% and 18.83±7.07,respectively.The difierences of SUV_(max) of T/WM,HIF-1α level and MVD between two groups were statistically significant (t=-5.921,z=-3.938,t=-6.745,all P＜0.05).(2)Among 41 gliomas,the strong positive expression of HIF-1α was observed in 8,mederate in 9,weak in 15 and negative expression was found in 9,SUV_(max) of T/WM and MVD increased with increasing HIF-1α level.The differences of SUV_(max) of T/WM and MVD among 4 different groups were statistically significant (F=7.41,P＜0.05).(3) The MVD of all gliomas was ranged from 9.76 to 94.52,which correlated with SUV_(max) of T/WM(r=0.759,P＜0.05).Conclusions The SUV_(max) of T/WM correlates with HIF-1α level and MVD in gliomas.Therefore,~(18)F-FDG PET provides preoperatively a noninvasive assessment of hypoxia or angiogenesis in human glionma.

Objective To study the effect of survivin anfisense oligonucleotides (ASODN)combined with quercetin on proliferation, apoptosis and cell cycle of a hepatocellular carcinoma cell line SSMC-7721 cells. Methods Human hepatocellular carcinoma cell line SSMC-7721 was cultured in vitro,and cells on logarithmic growth phase were used for this experiment. Cell proliferation was measured by MTT assay. The apoptotic rate and cell cycle were examined by flow cytometer (FCM). Morphological change of apoptotic cells were observed by fluorescent microscope. The expression of survivin gene was detected by the method of immunohistochemistry staining and RT-PCR on the mRNA and protein level. Results After sealing survivin gene with ASODN, the proliferation of SSMC-7721 cells was inhibited markedly. FCM analysis showed that there appeared an obvious apoptosis peak after transfection. The inhibitory effect of combined administration of survivin ASODN and quercetin on cell proliferation was much stronger than that of the single way. The result of immunohistochemical and RT-PCR assays showed that survivin ASODN and quercetin inhibited the expression of survivin gene. Conclusion Combined survivin ASODN with quercetin significantly inhibit cell proliferation, down-regulate survivin gene expression and induce the apoptosis of SSMC-7721 cells.

The goal of the study was to investigate the protective effect and mechanism of Artemisia argyi ethanol extract on chemotherapeutic vancomycin (VAN)-induced acute kidney injury (AKI).The acute kidney injury model of male ICR mice was induced by intraperitoneal injection (ip) of VAN.Thirty mice were divided into the blank group, model group, high dose group, middle dose group and low dose group, which were given medicine by gastric perfusion (ig).Serum levels of cystain C (Cys C), blood urea nitrogen (BUN) and serum creatinine (Scr) were measured, which could reflect renal function of mice.Serum oxidative stress and inflammation indices were also determined, including muscular dystrophy association (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), tumor necrosis factor-alpha (TNF-α), Interleukin-1β (IL-1β), and high-sensitive c-reactive protein (hs-CRP).In addition, hematoxylin-eosin staining (HE) was employed for measuring the damage of renal tissues and the content of apoptosis b-cell lymphoma-2 (Bcl-2), Bcl-2 assaciated X protein (Bax) and caspase-3 were measured too.All results showed that Artemisia argyi extract exhibits protective effect on chemotherapeutic VAN-induced AKI, whose mechanism could be related to the oxidative stress, inflammatory reaction and apoptosis.

The strain of Cunninghamella echinulata 9980 was first selected with high aminoacylase activity . In three submerged cultures, the aminoacylase activity in the mycelial cell was compared . A number of factors have effects on the resolution reaction. The results showed that, peptone culture gave the highest aminoacylase activity with 680U/g.The optium temperature,pH,and substrate concentration were 55℃, 7.0,and 0.2mol/L,respectively. The ions in the buffer lowered the activity,but the Co~(2+) in 10~(-4)~10~(-3 )mol/L was necessary for its activity.

The study is aimed to understand the resource and the current situation of the use of Cibotii Rhizoma and provide the basis for protecting and utilization. The method of literature survey, field survey and quality assessment were applied in the study. The results showed that all the Cibotii Rhizoma came from wild resource and was mainly founded in Fujian, Guangxi, Guizhou, Yunnan, Guangdong, Hunan, Jiangxi, Sichuan, Chongqing, Zhejiang, etc. It contains over 5 000 000 kg in the area which total is about 7 000 hm2. The annual output is over 850 000 kg. At present, there is no cultivated resources. Based on the investigation and market sampling analysis from various regions, the results showed that the quality of the collected crude drugs conformed with the regulations of the Chinese pharmacopoeia. However the qualification rate of decoction pieces of Cibotii Rhizoma in market was only 56.4%. At present, the resource of Cibotii Rhizoma could meet the needs of medinal uses. It is important to protect the wild resource which is less and less because of the environmental factors. It also need to make a standard of processing method to ensure the safety, and solve quality problem of the decoction pieces.
China ; Conservation of Natural Resources ; Drugs, Chinese Herbal ; analysis ; Ferns ; chemistry ; growth & development ; Quality Control ; Rhizome ; chemistry ; growth & development

Objective To assess the long-term surgical outcomes,patient's satisfaction and functional results of correction for patients with penile curvature by 16-dot minimal tension plication.Methods Between August 2004 and December 2010,92 patients with penile curvature underwent surgical correction of penile curvature by 16-dot minimal tension plication.Of them,86 patients (53 patients with congenital penile curvature and 33 patients with Peyronie's disease (PD)) were reached for long-term follow-up.Indications for operation included patients with 30 to 90 degrees of penile curvature,penile length greater than 10 cm and patients who had one or more symptoms,such as difficulty or inability to vaginal penetration and partner discomfort during sexual intercourse.Patients were excluded from study if they had severe penile curvature (greater than 90 degrees),hourglass deformities or large plaques.Preoperative evaluation included international index of erectile function 5 (IIEF-5) questionnaire score,pain on erection and physical examination.Patients were recommended daily low dose phosphodiesterase type 5 (PDE-5) inhibitors 2 weeks after surgery and maintained for 2 to 3 months.At follow-up,recurrence rate,complications,erectile function,patients and partner's satisfaction with the surgery were assessed.Results The mean time of follow-up was 79 months (range,35-107 months).Of 86 patients who had been reached for long-term follow up,82 patients (95.3％) and 77 partners (89.5％) reported overall satisfaction after the surgery.Complete penile straightening was achieved in 79 patients (91.8％).Slight residual curvatures were observed in 7 patients,but did not affect their sexual intercourse.Postoperative penile length shortening (median penile length loss 1.3 cm) was noted in 35 patients (40.7％),which did not affect their sexual intercourse.In addition,no de novo erectile dysfunction was observed,and sexual function was significantly improved evidenced by the IIEF-5 scores,especially in the patients who had received PDE-5 inhibitors for 2 to 3 months.There were no surgery-related complications,such as hematoma,urethral injury,or infection.Conclusion The longterm results demonstrate that correction of penile curvature using 16-dot plication is a simple and safe method to achieve cosmetic and functional satisfaction in patients with mild to moderate penile curvature.

OBJECTIVESBased on the immunologic character of Pichia pastoris yeast, a new therapeutic vaccine, whole recombinant yeast, was used to explore a new way to activate cell-mediated anti-viral immunity.

METHODSThe recombinant plasmids, pPIC9K/S and PIC9K/hsp(1-370)-S, were constructed by inserting the gene encoding HBsAg, HSP70 (1-370) -HBsAg into vector pPIC9K and then the recombinants were transfected into Pichia pastoris yeast,GS115, respectively. Then that recombinant yeast immunized BALB/C mice were detected for humoral and cellular immunity to HBsAg.

RESULTSRecombinant yeast successfully activated the humoral immunity to HBsAg in mice, but failed to activate the cellular immunity.

CONCLUSIONThe whole recombinant yeast can be used as vaccine, but need further study for optimal way of immunization.

Animals ; Female ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Pichia ; genetics ; Plasmids ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccines, Synthetic ; immunology

OBJECTIVETo observe the modulatory effect of subthalamic nucleus (STN) on activity of motor cortex during exhausting exercise.

METHODSElectrocorticogram (ECoG) and local field potentials (LFPs) recording techniques were applied simultaneously to observe the dynamic changes of oscillations in sensorimotor area and STN of rat during exhausting exercise.

RESULTSRats ran well initiatively with treadmill at the beginning of the exercise, about 45 min (45 +/- 11.5) later, movement capacity reduced. Corresponding electrical property showed that STN activity increased significantly while activity of cortex decreased significantly. Subsequently rats continued exercise with minor external stimulation utill exhaustion. Activity of ECoG reached the minimum under exhausting stations (P < 0.01), while the activity of LFPs changed insignificantly (P > 0.05).

CONCLUSIONDuring the exhausting exercise, the cortex activity was extensively depressed with the development of fatigue, while the activity of STN increased significantly at the early stage of fatigue, STN took part in the modulation of central fatigue through negative induction. And the increase of STN activity may be one of the key measures accounting for protective inhibition.

Animals ; Cerebral Cortex ; physiology ; Electrophysiological Phenomena ; physiology ; Male ; Neurons ; physiology ; Physical Conditioning, Animal ; physiology ; Physical Exertion ; physiology ; Rats ; Rats, Wistar ; Subthalamic Nucleus ; physiology

The purpose of study was to investigate the feasibility of the application of cationic propyl gallate (C-PG) as inducer of platelet aggregation for evaluating the platelet function of single-donor plateletpheresis and identifying the incidence of defective platelet function among donors. Experiments were as follows: 3 healthy volunteers' platelet aggregation induced by 100-300 micromol/L C-PG was determined by LG-PABER analyzer to observe the effect of C-PG concentration on platelet aggregation; 30 healthy volunteers' platelet aggregation before and 24 hours after administration of 200-400 mg acetylsalicylic acid (ASA) was examined after induction by 200 micromol/L C-PG for determining the cut-off value to discriminate platelet dysfunction donors; the platelet aggregation of 483 platelet donors was detected and the activated plasma clotting time (APCT) of donors who have deficiency in platelet aggregation was examined for investigating the incidence of defective platelet function among donors. The results showed that platelets were activated by C-PG induction in a dose dependent manner, when concentration of C-PG reached 200 micromol/L, the percentage of platelet aggregation was highest. It significantly decreased after 24 hours with ASA than that before the administration (P < 0.001), especially in 180 seconds induced by C-PG. If cut-off point was fixed on the platelet aggregation < 20% in 180 seconds, donors of platelet dysfunction can be selected effectively. 25 of defective platelet aggregation function among 483 donors were detected, and 11 out of 25 platelet dysfunction donors had the deficiency in procoagulant activity with prolonged APCT. It is concluded that C-PG as inducer of platelet aggregation is feasible to screen the platelet function of donors. Five percent of platelet donors has function defect examined by C-PG as inducer of platelet aggregation.
Antioxidants ; chemistry ; pharmacology ; Aspirin ; administration & dosage ; Blood Donors ; Blood Platelets ; cytology ; drug effects ; physiology ; Cations ; chemistry ; Humans ; Platelet Activation ; drug effects ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; administration & dosage ; Platelet Function Tests ; Platelet Transfusion ; Propyl Gallate ; administration & dosage ; chemistry ; Whole Blood Coagulation Time