Objective To study the effects of nitrobenzene on macrophage function and lymphocyte proliferation in mice. Methods ICR mice were divided into groups and treated with nitrobenzene by gavage,once a day,at doses of 2,20 and 200 mg/kg respectively,for 21 consecutive days.The mice were killed after 21 days of treatment and then the effects of nitrobenzene on the organs index,the maerophage function and the lymphocyte proliferation were determined.Results The maerophage function and the lymphocyte proliferation decreased as the increase of the dose of nitrobenzene.Conclusion The results of the present paper show that nitrobenzene may inhibit the immunity function of mice.

DNA barcoding is a technique in which species identification and discovery are performed by using short and standard fragments of DNA sequences. In this study, eleven species of Paris, including seven varieties, were sampled. Five chloroplast sequences, psbA-trnH, rpoB, rpoC1, rbcL, matK, and one nuclear marker, the second internal transcribed spacer (ITS2) of ribosomal DNA, were amplified and sequenced. The PCR amplification and sequencing efficiency, intra- and inter-specific divergence and barcoding gap were used to evaluate different loci, and the identification efficiency was assessed using BLAST1 and Nearest Distance methods. The ITS2 sequences in the studied samples of Paris were amplified and sequenced successfully using primers designed by our group, while matK showed low level in the amplification and psbA-trnH was difficult for sequencing because of over 800 bp and poly (A) structure. Analysis of the intra- and inter-specific divergence and barcoding gap showed ITS2 was superior to other loci. The ITS2 showed a much higher percentage of success (100%) in identification than other five loci, none of which indicated more than 50% except matK (52.9%). The 2-locus combination of rbcL+matK didn't improve ability of authentication. In addition, the rate of successful identification with ITS2 kept 100% when the samples were expanded to 67 samples of 29 species. In conclusion, ITS2 can be used to correctly identify medicinal plants of Paris, and it will be a potential DNA barcode for identifying medicinal plants of other taxa.

Partial nature of "promoting blood circulation and dieresis" of Salvia Miltiorrhizain was initially demonstrated by investigating the regulation effect of AQP2 expression in kidney of trauma blood stasis model rats with the Salvia Miltiorrhizain so as to provide guidance for its clinical deployment of administration. Random allocation was taken to averagely divide 30 SD rats into two groups: 10 rats in normal group and 20 rats in blood stasis syndrome group. Trauma blood stasis rat model was established by quantitatively beating. Then the rat model group was divided into model group and salvia group. After 7 days of treatment, the rat kidney AQP2 expression was detected, the content of urine AQP2 was compared and the damaged local muscle and kidney pathological changes were observed by immunohistochemical method and western blot method. Compared with that of the normal group, rats in model group had inflammatory cells infiltration, blood stasis and edema of the injured local muscles and up-regulated AQP2 expression, decreasing urinary output, and kidney tissues blood stasis and edema (P < 0.05). On the other hand, compared with that of the model group, those parameters of rats in salvia group were all decreasing except urine output (P < 0.05). Such result indicated that Salvia Miltiorrhiza can reduce trauma blood stasis rat content of urine AQP2 and down-regulated AQP2 expression in kidney tissue, so as to reduce the reabsorption of water by renal tubular and increase urine output. The promoting blood circulation effect of Salvia Miltiorrhizain can alleviate the degree of the damaged tissue edema and encourage urine drainage. This therapy is closely related to the effect of regulating AQP2 in kidney by salvia, so the purpose of this study by verifying "promoting blood circulation and diuresis" as the mechanism for the regulation effect of the salvia on AQP2 expression.
Animals ; Aquaporin 2 ; genetics ; metabolism ; Blood Circulation ; drug effects ; Diuresis ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Kidney ; blood supply ; drug effects ; metabolism ; physiopathology ; Kidney Diseases ; drug therapy ; genetics ; metabolism ; physiopathology ; Male ; Rats ; Salvia miltiorrhiza ; chemistry

Background and purpose:Researches had indicated that about over 85%of malignant tumors highly express telomerase activity. So telomerase has become one of the important methods in the research field of tumor diagnosis and treatment. Nowadays, several reports about malignant tumor which over expresses hTERT targeting imaging with radionuclide labeled hTERT ASON had been published. In these reports, high quality of pictures can hardly be acquired because of poor anatomical and spacial resolution in nuclear imaging itself. Accordingly, in this study, we developed a method of detecting human telomerase in vivo with magnetic resonance imaging (MRI) and evaluate its feasibility. Methods:Firstly, Uniformly phosphorothioate-modified human telomerase reverse transcriptase antisense oligonucleotide (hTERT ASON) was labeled with Gd3+ through the bifunctional chelator 1, 4, 7, 10-tetraazacyclododecane-N, N’, N’’, N’’’-tetraacetic acid (DOTA) and iv vitro experiments were performed to characterize the antisense probes (for biodistribution and cellular uptake, 99mTc-DOTA-ASON was used in stead of Gd-DOTA-ASON). Then Gd-DOTA-ASON was injected intraperitoneally in pulmonary adenocarcinoma A375 nude mice tumor-bearing BALB/c for in vivo imaging using 7.0 T Micro MRI periodically, tumors and their surrounding tissues were defined as region of interest (ROI) to calculate the signal to noise ratio (SNR) of tumor to muscle using Gd-DTPA as control. Finally, immunohistochemical analysis of telomerase activity of each xenograft was operated 2 days after imaging. Results:The binding efficiency of Gd-DOTA-ASON reached was as high as 65%(63.2±2.4, n=6). And it can maintain 61%in fresh human serum and normal saline at 37℃over 24 h;A375 cells showed an uptake of 8.5%when incubated with 99mTc-DOTA-ASON;In comparing with DOTA-ASON and Gd-DTPA, cells transected with Gd-DOTA-ASON had higher SI when performed MRI with T1WI. The hTERT-expressing xenografts were obviously enhanced by Gd-DOTA-ASON at 0.5-6 h after injection and the SNR can reach 2.37, whereas obvious enhancement only could be found within 2 h after injection of Gd-DTPA. Both labeled and non-labeled antisense probes can suppress the activity of telomerase of A375 cells either in vitro or in vitro. Conclusion:Our research offers proof that Gd-DOTA-ASON can be used as tumor specific targeting MR probe for diagnosing malignant tumors with high expression of telomerase.

Objective To study the expression and significance of Th17 cells from peripheral blood of patients with rheumatoid arthritis (RA). Methods Intracelluar flow cytomete detection of IL-17/IFN-γ and IL-17/IL-6 was established using anti-CD3/Anti-CD28/IL-23 as stimulators after isolation of untouched human CD4~+T cells from PBMC. There were three groups in the present study: ①healthy controls group; ② RA stable group; ③RA active group. Results The isolation of untouched human CD4~+T cells from PBMC was effective and its purity was over 90%. The percentage of intracelluar IL-17 in CD4~+ T cells from RA patients was increased significantly. Such percentage in active group (1.54±0.41) was higher than that of stable group (0.70±0.21, P<0.01) and both of them were higher than those of healthy controls (0.42±0.12, P<0.01). Under anti-CD3/Anti-CD28/IL-23 stimulation, the percentage of intracelluar IL-17 was also increased significantly(P<0.01). The porcentage of intracellular IFN-γ was similar to that of IL-17, while that of IL-6 was not significantly different. There is an correlation between IL-17 and IFN-γ or IL-6. Conclusion There is an abnormal expression of IL-17 and IFN-γ in human CD4~+T cells in RA patients, which is related to disease activity . Th17 cells may be used as a new marker for the assessment of RA activity.

Objective To investigate the mechanism of tumor necrosis factor-α (TNF)-α inhibiting osteo blastdifferentiation of mesenchymal stem cells (BMMSCs) in the pathogenesis of osteoporosis in the mouse model of systemic lupus erythematosus (MRL/lpr). Methods The femurs of MRL / lpr and C3He/HeJ mice were isolated, the bone structure were examined by hematoxylin-eosin (HE) staining. The proteins of TNF-α, NF-κB P50, bone morphogenetic protein -2 (BMP-2) and PSmad1/5/8 were measured by immunohistochemical stain. Bone marrow mesenchymal stem cells (BMMSCs) were isolated. After BMMSCs grew on the cover slips, the proteins on top of it were evaluated by immunohistochemistry stain. Moreover, the alkaline phosphatase (ALP) staining was employed for the measurement of the early osteogenic differentiation. BMMSCs together with hydroxyapatite were embedded subcutaneously in the nude mice and eight weeks later, the ectopic bone formation was evaluated. The recombinant human tumor necrosis factor receptor type Ⅱantibody fusion protein (etanercept) or normal saline was subcutaneous injected to the mice with lupus. After four weeks, the expression of these proteins was observed and the ectopic bone formation was investigated. Image-Pro plus 6.0 software was employed for imagine analysis, and Studentˊs t-test was used to test the differences between 2 independent groups. Results MRL/lpr mice showed decreased volume of cortex and the percentage of cortex to the volume of bone of MRL/lpr mice was significantly lower compared to control groups and with C3He/HeJ mice (13.96±0.25 vs 23.61±0.71, n=3, P<0.01). The protein levels of both TNF-αand NF-κB P50 on the femur of MRL/lprl mice were higher than those of the control group (0.643±0.051 vs 0.405±0.022, 0.917±0.023 vs 0.650±0.032, n=3, P<0.01). The expressions of BMP-2 on the femur of MRL/lpr mice were lower than those of the C3He/HeJ mice (0.52 ±0.03 vs 0.72 ±0.03, n=3, P<0.01). There was no difference in the expression of PSmad1/5/8 on the femur between the two groups by immunohistochemistry detection (1.264 ±0.021 vs 1.301± 0.044, n=3, P>0.05). The expressions of TNF-α and NF-κB P50 in BMMSCs of MRL/lprl mice were higher than those of the C3He/HeJ (0.184±0.021 vs 0.136±0.013, 0.132±0.021 vs 0.097± 0.014, n=3, P<0.01), while BMP-2 and PSmad were lower than those of the control group (0.128±0.013 vs 0.216±0.221, 0.115±0.023 vs 0.196±0.034, n=3, P<0.01). After 7 days of BMP-2 stimulation, the activities of ALP of BMMSCs from MRL/lprl mice were reduced detected by ALP staining and the osteoblast differentiation of these cells were decreased than BMMSCs from the control mice by HE and Masson staining. The percentage of the cortex to the volume of bone of the etanercept injection MRL/lpr mice was higher than that of the control group (21.8±1.0 vs 14.3 ±0.6, n=3, P<0.01). Moreover, the proteins of TNF-α and NF-κB P50 on the femurs of such injected mice were lower than those of the control group (0.540±0.024 vs 0.682±0.031, 0.857±0.023 vs 1.098±0.044, n=3, P<0.05), while the expressions of BMP-2 were higher than the control group (0.99±0.04 vs 0.85±0.04, n=3, P<0.05). There was no difference in the PSmad1/5/8 expression on the bone of the two group of lupus mice (0.88 ±0.08 vs 0.84 ±0.04, n=3, P>0.05). The ectopic bone formation of BMMSCs of the etanercept injected MRL/lpr mice was higher than that of the normal saline injected mice, however, it was lower than that of the C3He/HeJ mice. Conclusion TNF-α inhibits osteoblast differentiation of mesenchymal stem cells by depressing Smad signaling which may contribute to the osteoporosis of the lupus mice.

Objective To explore the clinical application value of neutrophil gelatinase-associated lipocalin(NGAL)which were tested by immunity transmission turbidity in early kidney injury after elective percutaneous coronary intervention.Methods A case-control study was conducted.All 201 stable angina pectoris and acute coronary syndrome patients undergone percutaneous coronary intervention in TEDA International Cardiovascular Hospital,during April to August 2013,were enrolled in this study.Before and 2 h,4 h,8 h,24 h,48 h after the operation,the plasma creatinine of the patient samples were tested by enzymic method.Before and 2 h,4 h,8 h,24 h after the operation,the plasma NGAL was tested by immunity transmission turbidity method.Before and 8 h,24 h after the operation,the urinary NGAL was tested by immunoturdimetric method.The data were compared between contrast induced nephrpathy (CIN) and non-CIN groups.For normal distribution of quantitative data,t test were used and for non-normal distribution of quantitative data,nonparametric rank and inspection were used.Results CIN occurred in 8 of 201 enrolled patients,the incidence was 3.98％.Receiver operating characteristic curve (ROC) analysis confirmed the diagnostic accuracy of the plasma NGAL in CIN,and the area under the curve(AUC) of 2 h plasma NGAL was 0.928,95％ CI 0.800-0.985,with the cut-off value NGAL as 109 ng/ml,the diagnostic sensitivity and specificity for CIN were 87.5％ and 100％ ;the AUC of 8 h plasma NGAL was 0.945,95％ CI 0.824-0.992,with the cut-off value NGAL as 96 ng/ml,the diagnostic sensitivity and specificity for CIN were 87.5％ and 87.5％ ;the AUC of 8 h urinary NGAL was 0.969,95％ CI 0.859-0.999,with the cutoff value NGAL as 91 ng/ml,the diagnostic sensitivity and specificity for CIN were 87.5％ and 100％.Conclusions The change of plasma and urinary NGAL is earlier to that of serum creatinine for the early diagnosis of CIN.It can be used as the predictor of early renal damage after elective coronary artery interventional.

Objective:To compare the analgesic effect between living rhino horn and rhino horn in mice and rats,and to explore the possibility of living rhino horn used as a substitute of rhino horn. Methods:The analgesic effect was compared using the body tor-sion method and the formaldehyde method in mice,and the hot plate method and the thermal sting imager method in rats. Results:Compared with the control group,the living rhino horn at the dose of 0. 35,0. 7 and 1. 4 g·kg - 1 could significantly prolong the incu-bation period of body torsion induced by acetic acid(P < 0. 05 or P < 0. 01),and significantly reduce the number of body torsion(P <0. 05 or P < 0. 01). The three dose groups(0. 35,0. 7,1. 4 g·kg - 1 )of rhino horn could significantly reduce the number of body tor-sion(P < 0. 05 or P < 0. 01). After the second dose and compared with the control group,the pain threshold of high dose group(1. 4 g·kg - 1 )of living rhino horn,high and middle dose groups(0. 7,1. 4 g·kg - 1 )of rhino horn was significantly prolonged(P < 0. 05 or P < 0. 01). Compared with the control group,three dose groups(0. 175,0. 35,0. 7 g·kg - 1 )of living rhino horn and rhino horn could significantly reduce the analgesic effect in mice induced by formaldehyde in the second phase(P < 0. 01). Compared with the control group,the changes of pain threshold before and after the administration in three dose groups(110,220,440 mg·kg - 1 )of liv-ing rhino horn and high dose group(440 mg·kg - 1 )of rhino horn was significantly increased(P < 0. 05 or P < 0. 01). Conclusion:Living rhino horn can be used as a substitute of rhino horn with promising analgesia effect.

Objective To study the protective effect of compound nitroglycerin gel on rats with skin ulcer wound and its action mechanism. Methods Skin ulcer modle of 54 rats was established.Then the rats were randomly divided into three groups (n=18 each), including model control group, Jin wan hong group, and compound nitroglycerin gel group. Wound healing process and healing time were recorded.At the day 7 and 14 after the model was established, the number of fibroblasts and new blood capillaries of granulation tissue from center of the wound were measured,and RT-PCR was used to detect mRNA expression levels of VEGF, Ang1 andHIF-1α. Results As compared with model control group [(24.17±5.91) days], healing time of skin in the compound nitroglycerin gel group was significantly shorter [(14.67±3.76) days, P<0.01].The numbers of fibroblasts (61.20±7.56) and new blood capillaries (9.35±1.43) were increased, and mRNA expression levels of VEGF (1.692±0.196), HIF-1α (1.527±0.174) and Ang1 (1.548±0.203) were remarkably up-regulated on 7th day (P<0.05 or P<0.01).While on 14th day, the numbers of fibroblasts (28.00±5.96) and new blood capillaries (4.20±1.30) were decreased and the mRNA expression levels of VEGF (1.156±0.123), HIF-1α(1.021±0.105) and Ang1 (1.034±0.134) were significantly down-regulated (P<0.05 or P<0.01) . Conclusion Compound nitroglycerin gel can treat skin ulcer wound via regulating the numbers of fibroblasts, new blood capillaries and VEGF/Ang1/HIF-1α signal transduction pathway.

Objective To investigate the value of high-resolution magnetic resonance imaging (HRMRI)in diagnosis of extramural vascular invasion (EMVI)of rectal cancer.Methods 33 patients with rectal cancer were reviewed preoperatively.The MRI findings of EMVI of all cases were scored and compared with the postoperative pathological results.Results The MRI EMVI scores were consistent with histopathology findings (k=0.324,P=0.039).The accuracy rate of MRI in diagnosis of EMVI was 66.7% (22/33).The MRI EMVI scores rose up with increased pT stage,meanwhile there was a high correlation between both (r=0.546).The percentage of MRI EMVI positive number was increased with elevated pT stage,and there was also a high correlation between both (r=0.469). ROC curve showed that MRI EMVI scoring was an effective method in diagnosis of rectal cancer EMVI (AUC=0.757).Conclusion HRMRI is a valuable method in diagnosis of EMVI of rectal cancer.