Objective To observe the effects of mycophenolate mofetil (MMF) on the differentiation and proliferation of Helper T cells 17 (TH 17),so as to reveal its role and the possible mechanism in inducing immunological suppression.Methods Sixteen Balb/c mice of SPF level aged 8 weeks were randomly divided into two groups:MMF group and control group,with 8 mice in each group.In MMF group,the mice received intragastric administration of MMF (40 mg·kg-1· day-1 ),and those in control group received intragastric administration of identical volumetric saline every day.After three weeks,peripheral blood was collected and spleen cells were prepared.Flow cytometry was used to determine the proportions of CD4+ TH 17 and CD4+ CD25+ Tregs,then the ratio of TH 17/Tregs was calculated,and the concentrations of interleukin-1 7 (IL-1 7) and interleukin-23 (IL-23) in serum were measured by ELISA.Results The proportion of CD4+ TH 17 in the peripheral blood and spleen was (1.95 ± 0.08) and (2.42 ± 0.06) in MMF group,and (3.19 ± 0.07)％ and (4.21 ± 0.25)％ in control group,respectively.There were significant differences between the two groups (P ＜0.05).Meanwhile,the ratio of TH 17/Tregs in MMF group,both in the peripheral blood and spleen,was significantly decreased as compared with the control group (P＜0.05).The concentration of IL-17 in MMF group was lower,but that of IL-23 in MMF group was higher than in the control group (P＜0.05).Conclusion MMF could obviously suppress the differentiation and proliferation of CD4+ TH 17 in vivo,reduce the ratio of TH17/Tregs and the IL-17 secretion,thus facilitate the induction of immune tolerance.

Objective To investigate the protective mechanism of protein kinase C(PKC)and calcium sensing receptor(CaR)in ischemia preconditioned rat hearts.Methods Using cell culture method,in vitro cultured inhibitor(IPC+CaRI).Apoptosis was detected using TUNEL and Hoechst33342 cell viability was detected by MTT,the protein expression of easpase-12,calpain and CaR in endochylema were detected using Wedtetm blot.ResultsIn I/R group nucleus was shrank,big blue,chromatin concentrated,apoptotle body appeared.Other groups haddifferent fluorescence intensity varying degree,IPC+PKCI+CaRS group had more big blue nucleus.Myocardialcell viability and apoptotic rate,I/R group[(62.99±0.65)%,(19.13±0.87)%],IPC group[(78.67±0.37)%,(14.21±0.74)%],IPC+PKCI group[(71.09±0.52)%,(20.46±0.81)%],IPC+PKCI+CaRS group(66.10±0.75)%,(24.89±1.43)%],IPC+CaRS group[(69.56±0.44)%,(21.64±0.77)%],IPC+CaRI group(85.81±0.60)%,(13.12±0.69)%],all had a difference(P＜0.05 or＜0.01)compared with C group[(100.00)%,(6.02±0. 31)%].Western blot identified that CaR expression in IPC+PKCI and IPC+CaRS,IPC+PKCI+CaRS groupswas more than that in IPC and IPC+CaRI groups;easpase-12 had more active fragment(60×103)in I/R,IPC+CaRS,IPC+PKCI+CaRS groups;ealpain expressions in I/R,IPC,IPC+PKCI,IPC+PKCI+CaRS,IPC+CaRSgroups were higher than those in C and IPC+CaRI,I/R group was the highest one,C group the second,IPC+CaRI the third.Conclusion The interaction of PKC and CaR can reduce the intracellular Ca2+ from sarcoplasmicreticulum thus provide a protection.

Summary: In order to investigate the effects of aconitine on [Ca2+] oscillation patterns in cultured myocytes of neonatal rats, fluorescent Ca2+ indicator Fluo-4 NW and laser scanning confocal microscope (LSCM) were used to detect the real-time changes of [Ca2+] oscillation patterns in the cultured myocytes before and after aconitine (1.0 μmol/L) incubation or antiarrhythmic peptide (AAP) and aconitine co-incubation. The results showed under control conditions, [Ca2+] oscillations were irregular but relatively stable, occasionally accompanied by small calcium sparks. After incubation of the cultures with aconitine, high frequency [Ca2+] oscillations emerged in both nuclear and cytoplasmic regions, whereas typical calcium sparks disappeared and the average [Ca2+] in the cytoplasm of the cardiomyocyte did not change significantly. In AAP-treated cultures, intraecllular [Ca2+] oscillation also changed, with periodic frequency, increased amplitudes and prolonged duration of calcium sparks. These patterns were not altered significantly by subsequent aconitine incubation. The basal value of [Ca2+] in nuclear region was higher than that in the cytoplasmic region, in the presence or absence of drugs, the [Ca2+] oscillated synchronously in both the nuclear and cytoplasmic regions of the same cardiomyocyte. It was concluded that although oscillating strenuously at high frequency, the average [Ca2+] in the cytoplasm of cardiomyocyte did not change significantly after aconitine incubation, compared to the controls. The observations indicate that aconitine induces the changes in [Ca2+] oscillation frequency other than the Ca2+ overload.

The effect of triptolide on proliferation and apoptosis of human multiple myeloma RPMI-8226 cells in vitro,as well as the roles of nuclear factor-kappa B (NF-κB) and IκBα was investigated.The effect of tritptolide on the growth of RPMI-8226 cells was studied by MTT assay.Apoptosis was detected by Hoechest 33258 staining and Annexin V/PI double staining assay.The expression of NF-κB and IκBα was observed by Western blot and confocal microscopy.The results showed that triptolide inactivated NF-κB apoptotic pathway in human multiple myeloma RPMI-8226 cells.Triptolide at nM range induced proliferation inhibition in a dose- and time-dependent manner and apoptosis in a dose-dependent fashion in RPMI-8226 cells.Besides,we observed the inhibition of NF-κB/p65 in the nuclear fraction was correlated with the increase in the protein expression of IκBα in the cytosol.These results suggested that triptolide might exhibit its strong anti-tumor effects via inactivation of NF-κB/p65 and IκBα.

Objective To investigate the effects of selective cyclooxygenase 2 inhibitor on the hyperplasia of parathyroid glands from uremic rats. Methods Sixty-five 5/6-nephrectomized (Nx) and fifteen sham operated rats were assigned to 4 groups: (1)Sham group (n=14):shamoperated +normal phosphate diet (P 0.8％,Ca 1.2％)； (2) Nx-HP group (n=17):Nx+high phosphate(HP) diet (P 1.2％,Ca 1.2％)； (3)Prophylactic COX2 inhibition group (Prey group,n=18):Nx+HP+celecoxib 100 mg· kg-1·d-1 for 3 months； (4)Therapeutic group (Ther group,n=18):Nx+HP+celecoxib 100 mg·kg-1·d-1 starting at the second month of the 5/6 nephrectomy.At the end of 3 month,blood,urine and parathyroid samples were collected.The expressions of COX2 and PCNA were determined by immunohistochemistry,Western blotting and real-time PCR. Results All of the Nx rats fed with high phosphate diet for 3 months manifested progressively increasing serum creatinine,serum iPTH as well as augmentation of parathyroid gland volume,suggesting that secondary parathyroid hyperplasia animal model was established successfully.Celecoxib significantly decreased serum iPTH levels [Sham (34.77±0.83),Nx-HP(100.73±4.35),Prey (87.36±2.18),Ther (87.47±1.76) ng/L,P＜0.05],the size of the parathyroid glands in Nx rats [Sham (0.461±0.089),Nx-HP (2.436±0.372),Prey (0.987±0.254),Ther (1.27±0.305) mm2/kg,P＜0.05] and PCNA expression in PG determined by Western blotting (decreased to 52.91％ in Prev group and 34.68％ in Ther group respectively,P＜0.05).No significant difference was observed between the two COX2 inhibition groups.The levels of COX2 expression in parathyroid gland were greatly increased in three Nx groups compared with that in sham group (2.47-fold in Nx-HP,2.34-fold in Prey group,3.04-fold in Ther group,P＜0.05).COX2 inhibitor had no effects on COX2 expression in PGs.Real-time PCR analysis demonstrated the same trends of mRNA expression of COX2 and PCNA in PGs of rats. Conclusion Selective inhibition of COX2 may help to suppress the hyperplasia of parathyroid glands in uremic rats.

Objective The study aimed to investigate the relationship between arrhythmia occurrence and nerve remodeling of thoracic spinalcord 1-5 nerves as well as myocardial electrophysiological remodeling in a metal stress rat model.Methods Thirty SD rats (weight 180-250 g)were randomly divided into control group (n = 10),stress group (n = 10) and fluoxetine group (n = 10,10 mg/kg i.p.for 3 weeks).Stress model (given by unpredicted chronic mild stress) was established according to Cronli's protocol.Following parameters were observed:( 1 ) ECG waveform change and arrhythmias ; (2)tissue field action potential duration (FAPD) of thoracic spinal cord 1 - 5 and cardiac tissue mapped by microelectrode arrays ( MEA ) technique; ( 3 ) myocardial growth-associated protein ( GAP-43 ),tyrosine hydroxylase (TH),choline acetyltransferase (CHAT) distribution observed by immunofluorescence and confocal laser scanning microscope (LSCM).ResultsThree weeks later:(1) The body weight,food intake,consumption of sugar water,the horizontal and vertical movement score,cleaning action of rats were significantly decreased,and fecal grains significantly increased,P-wave,P-R interval,QRS-wave and Q-T interval were significantly prolonged and heart rate was significantly reduced in stress group compared with control group ( all P ＜0.05).Incidence of ventricular premature beat was 80％ in stress group and 0％ in control group (P ＜ 0.05 ).The FA PD of thoracic spinal cord 1 -5 nerves [ ( 144.25 ± 12.63 )ms vs (79.56 ±8.01 )ms ] and of cardiac tissue [ LA ( 122.43 ± 19.34)ms vs (92.59 ± 7.61 )ms,RA ( 149.89 ± 14.68 )ms vs (105.18 ± 15.94) ms,LV(162.62 ± 7.04) ms vs (110.45 ±6.92) ms,RV (152.21 ± 30.49) ms vs ( 131.06 ± 12.04) ms ] were significantly prolonged,FAPD dispersion (FAPDd) significantly increased [ thoracic spinal cord 1 - 5 ( 13.3 ± 9.11 ) ms vs ( 9.36 ± 7.01 ) ms ] in stress group compared with the control group.Disarrangement of myocardial cells,proliferation of collagen fiber,infiltration of neutrophil and lymphocytes in the cardiac tissue were also observed and distribution of GAP-43,TH and CHAT was significantly increased in stress group.(2) All these changes could be partly reversed by the treatment with fluoxetine.Conclusion Metal stress induced cardiac autonomic nerve and myocardial electrophysiological remodeling and ventricular arrhythmia in rats which could be significantly attenuated by fluoxetine in this model.