The effects of Rhynchophylline (Rhy) and Isorhychophyiline (Isorhy), the alkaloids abstracted from the Chinese traditional herb Uncaria rhynchophyllia (Miq) Jackson, on the 45Ca-influx and efflux were investigated in rabbit aorta. Both Rhy and Isorhy (10 ?mol? L-1) inhibited the 45Ca-influx induced by high K+(77. 0 mmol ? L-1), but neither significant-ly influenced the 45Ca-influx and efflux induced by noradrenaline (10 ?mol ? L-1). The results suggest that these alkaloids block the Voltage-dependent calcium channel.

AIM: To establish the method for determination of gentiopicrin, palmatine hydrochloride and berberine hydrochloride in Ertong Ⅱ Oral Liquid (Cortex Phellodendri, Radix Gentianae, Rhizoma Anemarrhenae, etc.) by RP HPLC. METHODS: The column was a Diamonsil TM C 18 column (250mm?4.6mm, 5?m); the mobile phase was CH 3CN H 3PO 4 C 6H 15 N H 2O; The ultraviolet detection was set at 270nm; the flow rate was 1mL?min -1 . RESULTS: The linear ranges of gentiopicrin, palmatine hydrochloride and berberine hydrochloride were 72.8～728?g?mL -1 ( r=0.9998 ), 7.55～75.5?g?mL -1 ( r=0.9998 ), 12.45～124.5?g?mL -1 ( r = 0.9999 ), respectively. The average recoveries ( n =6) of gentiopicrin, palmatine hydrochloride and berberine hydrochloride were 98.17%, 98.78%, 98.60%; RSD were 1.60%, 1.35%, 1.25%, respectively. CONCLUSION: The method is accurate, reproducible and highly selective, thus suitable for quality control of Er Tong Ⅱ Oral Liqual.

Objective To detect apoptosis of transplanted lung adenocarcinoma cells in nude mice after 125I brachytherapy by 99Tcm-Annexin V combined with diffusion weighted MRI (MR-DWI).Methods Twenty-five BALB/c-nu nude mice models subcutaneously transplanted with A549 cells were divided into experimental group (EG,n=13) and control group (CG,n=12) by random number table method.One 125I seed with apparent activity of (24.8±6.3) MBq was implanted into each mouse in EG,while CG underwent cold seed implantation.Both of 99Tcm-Annexin V imaging and MR-DWI were performed within 7-10 d after brachytherapy,then all mice were sacrificed and tumor cell apoptosis was detected by terminal oxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL) immunofluorescence,Survivin expression was assayed by SP.Two-sample t test,x2 test and Pearson correlation analysis were used for data analysis.Results Positive rate of cell apoptosis by 99Tcm-Annexin V imaging was 69.2％(9/13) and 8.3％(1/12) respectively in EG and CG (x2 =12.73,P＜0.01).The uptake ratio of 99Tcm-Annexin V,apparent diffusion coefficient(ADC) value and apoptosis index(M) of the tumor in EG were 2.91±0.85,(2.03±0.44)×10-3 mm2/s and (49± 18) ％,respectively,which were significantly higher than those of CG (1.26 ± 0.37,(1.29 ± 0.21) ×10-3 mm2/s and (11±4)％ respectively,t=5.930,5.452,7.606,all P＜0.05).Survivin expression in EG and CG was (46± 13) ％ and (15±7) ％ respectively (t =5.158,P＜0.05).The value of ADC was correlated with AI and uptake ratio(r=0.756,0.788,both P＜0.05).Uptake ratio was correlated with AI (r=0.754,P＜0.05),while Survivin expression was negatively correlated with AI (r =-0.772,P＜0.05).Conclusions Down-regulation of Survivin expression may play an important role in apoptosis induced by 125I brachytherapy.99Tcm-Annexin V combined with MR-DWI could effectively evaluate apoptosis of lung adenocarcinoma cells in a non-invasive way,thus it might be helpful in evaluation of early efficacy of 125I brachytherapy.

OBJECTIVE:To screen Gongshisong's active sites for anti-sports fatigue. METHODS:Gongshisong extract was prepared with 80% ethanol extraction technology,and extracted with petroleum ether,chloroform,ethyl acetate and n-butyl alco-hol after dispersed with water to obtain the extract. 70 mice were randomly divided into blank control group(1% sodium carboxy-methylcellulose,CMC-Na),positive control group [Rhodiola wallichiana capsules,590 mg/(kg·d)],petroleum ether,chloro-form,ethyl acetate and n-butyl alcohol extracts and aqueous layer of Gongshisong groups(TS,TL,TY,TZ,TW groups). Gong-shisong extracts groups was given relevant medicine 2.5 g(crude drug)/(kg·d),ig,for consecutive 7 days. Exhaustion time of bur-den swimming test was detected. 70 mice were grouped according to above method,and the contents of liver glycogen,muscle gly-cogen and the coefficient of liver were tested in mice. 80 mice were grouped according to above method,and model group was es-tablished additionally(1% CMC-Na). The contents of lactic acid(LA),creatine kinase(CK)and urea nitrogen(BUN)in serum of mice were determined after 90 minutes of unburden swimming. RESULTS:Compared with blank control group,exhaustion time of burden swimming mice in TS,TY and TZ groups prolonged;the content of liver glycogen increased in TY,TZ and TW groups;the content of muscle glycogen increased in TS and TW groups;the contents of BUN,LA and CK in mice increased in model group (P<0.05 or P<0.01). Compared with model group,the serum content of BUN in mice decreased in TS and TY groups;that of LA in mice decreased in TZ and TW groups;that of CK in mice decreased in TS group (P<0.05 or P<0.01). CONCLUSIONS:The petroleum ether and n-butanol extract site and water layer of Gongshisong are good anti-fatigue active sites.

Objective To evaluate the role of c-Jun N-terminal kinase (JNK) signaling pathway in isoflurane (ISO) preconditioning against cerebral ischemia/reperfusion (I/R) injury and investigate the relationship between JNK signaling pathway and apoptosis.Methods Global cerebral I/R models were made by 4-artery occlusion technique.Forty male SD rats of clean grade were divided into sham operation group (S group),I/R injury group (I/R group),SP600125 (an inhibitor of JNK signaling pathway) + I/R group (SP + I/R group),ISO preconditioning group (ISO group),and ISO preconditioning + SP600125 group (ISO + SP group) according to the random number table.Preconditioning protocol was successive inhalation of 15 g/L ISO for 5 days,1 h/d.I/R was induced at 24 hours after the end of preconditioning.Brain tissues were harvested at 72 hours later to take histomorphological examination by HE staining as well as detect apoptosis of hippocampal nerve cells by TUNEL method,expression of caspase-3 in hippocampal nerve cells by immuno-histochemistry,and expression of protein p-JNK in hippocampal tissues by Western blot.Results Compared with S group,brain injury score,apoptosis ratio of nerve cells,and expression of caspase-3 were significantly increased in the other groups (P ＜ 0.05).Moreover,p-JNK protein had a higher expression in IR group than in S group (P ＜ 0.05),but no significant difference was observed in SP + I/R group,ISO group,and ISO + SP group as compared with S group (P ＞ 0.05).Compared with I/R group,brain injury score,apoptosis ratio of nerve cells,expression of caspase-3,and expression of p-JNK protein were all declined in SP + I/R group,ISO group,and ISO + SP group (P ＜ 0.05).Moreover,brain injury score,apoptosis ratio of nerve cells,and expression of caspase-3 had further decline in ISO + SP group as compared with SP + I/R group and ISO group (P ＜ 0.05),but the difference in expression of p-JNK protein was insignificant among the three groups.Compared with SP + I/R group,no significant changes of each index were found in ISO group.Conclusion ISO preconditioning alleviates cerebral I/R injury through down-regulating expression of caspase-3 and inhibiting JNK signaling pathway.

Objective:To analyze the association between gene single nucleotide polymorphisms ( SNP) of methylenetetrahydrofo-late reductase ( MTHFR) gene and lower extremity atherosclerotic disease ( LEAD) . Methods:The clinical data and peripheral blood were collected from 384 participants (224 LEAD cases and 160 normal controls) from Han population of Minnan Fujian. LEAD was detected with ankle brachial index ( ABI) , toe brachial index ( TBI) , color Doppler ultrasonic examination and the other imaging stud-ies. The SNP genotypes including rs1801133, rs1801131, rs2274976, rs4846048, rs3737966, rs1537515, rs4846049, rs3834044, rs13306561 and rs3737964 in the MTHFR gene were detected by matrix-assisted laser desorption ionization-time of flight ( MALDI-TOF) . Results:The genotype distributions of the ten loci were in accordance with Hardy-Weinberg equilibrium. There were 37 obvi-ous linkage disequilibrium, including the association between rs4846048 and rs3737966 (D′>0. 9) and so on. There were significant differences (P=0. 02) in GCCTCGGAAT haplotypes of MTHFR gene groups between LEAD cases and the normal groups. The results from chi-square test of allele frequencies suggested rs1801131 (OR=1. 287),rs4846048 (OR=1. 844,P=0. 02), rs3737966(OR=1. 339),rs4846049 (OR=1. 314) and rs3737964 (OR=1. 522). Significant differences (P<0. 05) were observed between LEAD and the normal groups in Cochran- Armitage trend test and Dominant gene action test of rs4846048. Conclusion: The SNP of rs1801131,rs4846048,rs3737966,rs4846049 and rs3737964 might be associated with the susceptibility of LEAD,and rs4846048 gene mutation might serve as a risk factor for LEAD in the community-based population.

OBJECTIVETo explore the effects of acrylonitrile on T lymphocyte subsets, expression of toll-like receptor 4 and related cytokines in rats.

METHODSSixty-four Sprague-Dawley rats were randomly divided into 4 female groups and 4 male groups, and there were 8 rats in each group. Rats in each group were respectively given a single dose of 0, 5, 10 and 20 mg/kg acrylonitrile by gavage, once a day, 5 days a week, for 13 weeks. Blood and spleen T lymphocyte subsets was detected by flow cytometry, the mRNA expression of TLR4, IL-1β and TNF-α was analyzed by real-time quantitative PCR, the protein expression of TLR4 was evaluated by Western blot.

RESULTSCompared with control group, the percentages of blood CD3, CD4 T cells in 20 mg/kg female group and CD4/CD8 ratio in 5, 10 and 20 mg/kg female groups was significantly decreased, CD8 T cells in 20 mg/kg group was significantly increased (P < 0.05 or P < 0.01), blood CD3 T cells in 5 mg/kg male group, CD4 T cells and CD4/CD8 ratio in 20 mg/kg male groups were lower than that of control group, CD8 T cells in 20 mg/kg make group was significantly in oreased (P < 0.05 or P < 0.01). Spleen CD4, CD8 T lymphocyte percentages and CD4/CD8 ratio in 20 mg/kg female group decreased significantly, CD8 T cells in 20 mg/kg male group was significantly increased (P < 0.05 or P < 0.01), spleen CD3, CD4, CD8 T cells in 20 mg/kg male group and CD4/CD8 ratio in 10, 20 mg/kg male groups was also significantly decreased, CD3 T cells in 20 mg/kg and CD8 T cells in 10, 20 mg/kg male groups were significantly increased (P < 0.05 or P < 0.01) (TLR4 mRNA was lower expressed in 5, 10 and 20 mg/kg male groups and 10 mg/kg female group (P < 0.05 or P < 0.01), and TLR4 protein in 5 mg/kg female group and 20 mg/kg male group was significantly lower than control group (P < 0.05). The expression level of IL-1β mRNA was significantly decreased in 5, 10 and 20 mg/kg female group and 5, 10 mg/kg male group (P < 0.05 or P < 0.01), TNF-α mRNA was lower expressed in 10, 20 mg/kg female groups and 5, 10 mg/kg male groups (P < 0.01).

CONCLUSIONAcrylonitrile may lead to the changes of CD3, CD4, CD8 T lymphocyte percentages and CD4/CD8 ratio in rat blood and spleen, and also significantly effected the expression level of TLR4 mRNA and protein together with the secretion of IL-1β, TNF-α. This may cause effects on the cellular immune function.

Acrylonitrile ; toxicity ; Animals ; Female ; Interleukin-1beta ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; T-Lymphocyte Subsets ; drug effects ; immunology ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the effects of benzene poisoning on the expression of multidrug resistance 1 (MDR1) gene and P-glycoprotein (P-gp) in the bone marrow mononuclear cells (BMMNCs) of C57BL/6 mice.

METHODSC57BL/6 mice were randomly divided into control group (n = 24), low-dose group (n = 24), medium-dose group (n = 24), and high-dose group (n = 24) to receive corn oil, 25 mg/kg benzene, 50 mg/kg benzene, or 100 mg/kg benzene by gavage, once daily, 5 days/weeks, for 4 weeks. The mice were sacrificed on day 12, 26, or 29 of poisoning. Peripheral blood routine test was performed; real-time quantitative PCR was used to measure the MDR1 gene expression in BMMNCs; Western blot was used to measure the P-gp expression in BMMNCs.

RESULTSOn day 12, the red blood cell count and hemoglobin level in the high-dose group were significantly lower than those in the control group, low-dose group, and medium-dose group (P < 0.01 or P < 0.05). On day 26, the white blood cell count in the high-dose group was significantly lower than those in the control group, low-dose group, and medium-dose group (P < 0.01 or P < 0.05). At each time point, the mRNA expression of MDR1 gene in the low-dose group, medium-dose group, and high-dose group was significantly lower than that in the control group (P < 0.01). On day 26, the P-gp expression in the high-dose group was significantly lower than those in the control group, low-dose group, and medium-dose group, and the P-gp expression in the medium-dose group was significantly lower than that in the low-dose group (P < 0.01 or P < 0.05). On day 29, the P-gp expression in the low-dose group, medium-dose group, and high-dose group was significantly lower than that in the control group (P < 0.05).

CONCLUSIONBenzene poisoning can affect the expression of MDR1 gene and P-gp, which may be one of the mechanisms of benzene hematotoxicity.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Animals ; Benzene ; toxicity ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Drug Resistance, Multiple ; genetics ; Male ; Mice ; Mice, Inbred C57BL ; Monocytes ; cytology ; drug effects ; metabolism

Objective To study the relevance between the polymorphisms and haplotype of multidrug resistant gene (MDR1) and the ratio of blood concentration/dosage in individuals treated by tacrolimus in stable phases post renal transplanta-tion ,and to provide data for personalized FK-506 administration .Methods The MDR1 C1236T ,G2677T/A and C3435T gen-otypes of 104 renal transplantation patients were determined by PCR followed sequencing method .The blood concentration of tacrolimus was detected by EMIT method .The differences in concentration/(dose × weight) (C/D) ratios were compared a-mong all of the genotype groups treated by tacrolimus .Results In the 104 renal transplantation recipients ,the frequency of MDR1 C1236T ,G2677T/A and C3435T mutation alleles was 56 .73% ,55 .77% and 33 .17% ,respectively .A correlation was revealed among SNP of MDR1 C3435T ,TTT haplotype and the C/D ratios (P<0.05) .In patients of CYP3A5*3*3 ,A cor-relation was also found among TTT haplotype and the C/D ratios (P< 0.05) .No difference was found among the MDR1 C1236T ,G2677T/A ,CGC haplotype and the C/D ratios (P>0.05) .Conclusion It is demonstrated that genetic polymor-phisms of MDR1 C3435T and TTT haplotype are correlated with C/D ratio of tacrolimus in Chinese renal transplant patients in stable stage .These findings affect the dose-adjusted concentration of tacrolimus individually .

The purified elementary bodies of C. pneumoniae TW-183 were used for immunization of male BALB/c mice, the spleen cells of these mice were fused with SP2/0 cells and the hybrid cells were cloned by limiting dilution. One clone that secreted the C. pneumoniae monoclonal antibody (Cpn-McAb) stably was obtained finally. The Cpn-McAb belonged to IgG2b class and anti-Cpn-MOMP; the outcome of micro-immunofluorescence showed its weak cross reaction with the C. psittaci elementary body but it has no cross reaction with C. trachoma elementary body. It has the same speciality of the imported Cpn-McAb. For the evaluation of Cpn-McAb, the peripheral blood mononuclear cell specimens of 454 patients were detected by self-made Cpn-McAb and imported Cpn-McAb at the same time. The positive rates of Cpn-antigen were 53.3% for self-made Cpn-McAb and 52.6% for imported Cpn-McAb,showing high concordance between them (Kappa=0.714). The results showed that self-made Cpn-McAb has almost the same high specificity and sensitivity as imported Cpn-McAb, so the self-made Cpn-McAb may replace imported Cpn-McAb to detect Cpn specific antigen and be helpful to diagnosing and treating the clinical diseases associated with Cpn infection.
Animals ; Antibodies, Bacterial ; immunology ; Antibodies, Monoclonal ; biosynthesis ; genetics ; Antibody Specificity ; Chlamydia Infections ; diagnosis ; microbiology ; Chlamydophila pneumoniae ; immunology ; Hybridomas ; secretion ; Male ; Mice ; Mice, Inbred BALB C