OBJECTIVE:To determine the contents of heavy metals in commercial Chinese crude drugs and to provide reference for the monitoring of heavy metals in the crude drugs.METHODS:UV spectrophotometry was established to determine the contents of heavy metals in 6 vegetable drugs(Panax quinquefolium,Panax ginseng,Lycium barbarum,Radix et Rhizoma Glycyrrhizae,Bulbus Fritillariae Cirrhosae and Salvia miltiorrhiza)and 2 animal-source drugs(Scolopendra subspinipes mutilans and Bombyx Batryticatus).RESULTS:The linear range of heavy metal plumbum(reference substance)was 0.01～0.50 ?g?mL-1(r=0.997 7),and the average recovery rate was 99.49%(RSD=1.41%).The contents of heavy metals in Panax quinquefolium,Panax ginseng,Lycium barbarum,Radix et Rhizoma Glycyrrhizae,Bulbus Fritillariae Cirrhosae,Salvia miltiorrhiza were 26.13,28.96,26.99,29.98,26.63 and 28.06 ?g?g-1 respectively,and the contents of Scolopendra subspinipes mutilans and Bombyx Batryticatus were on the high side(47.62 and 59.24 ?g?g-1 respectively).CONCLUSION:The method is easy,highly accurate,and it can be used for detection of heavy metals in Chinese crude drugs.

Purpose To observe the curative effect of special acupuncture techniques on female urethral syndrome and its relationship with the course of treatment.Method Four abdominal and four sacral empirical points were acupunctured with special techniques and electricity. A difference in curative effect was investigated between different numbers of treatments. Results The clinical cure rate reached 15.2% just after ≤ 10 (7.3 ± 1.3) treatments. The curative effect was significantly better after 20-40 (32.1 ± 5.8) treatments than after ≤ 10 (7.3± 1.3) treatments (χ2 = 10.086, P ＜0.05). The clinical cure rate reached 43.5% in the former. Conclusion Special acupuncture techniques have a good clinical effect on female urethral syndrome. The curative effect improves with an increase in the number of treatments.

Objeetive To explore the value of antisense imaging of 99Tcm-labeled ASON targeting mouse double minute 2(MDM2) mRNA for the diagnosis of human prostate cancer.Methods The ASON targeting MDM2 mRNA and the mismatched oligonucleotide (ASONM) were synthesized and radiolabeled with 99Tcm using the bifunctional chelator HYNIC.The labeling efficiency and radiochemical purity were investigated.Animal models of nude mice bearing human prostate cancer LNCaP were established and divided into 3 groups with 10 mice in each group.99Tcm-HYNIC-ASON,99Tcm-HYNIC-ASONM (study groups) and 99TcmO4-(control group) were injected at the dose of 7.4 MBq through the tail vein,respectively.Tumor imaging was acquired with SPECT and the tumor-to-muscle (T/M) ratio was measured.The data was compared by one-way analysis of variance.Results The labeling efficiencies of ASON and ASONM were (65.15± 2.05) ％ and (64.93±2.18) ％,respectively.Their radiochemical purity was greater than 90％.At 1,4 and 10 h post injection,the T/M ratios of 99Tcm-HYNIC-ASON group were 3.217±0.125,3.749± 0.201 and 4.028±0.186,and those of 99Tcm-HYNIC-ASONM group were 1.579t0.128,1.715±1.140 and 1.683±0.139,and control group 2.146±0.132,1.847±0.124,1.528±0.152,respectively.The T/M ratios in control group and 99Tcm-HYNIC-ASONM group were significantly lower than those in 99Tcm-HYNICASON group at 1,4 and 10 h,respectively (F=213.37-235.41,t=3.527-4.738; all P＜0.01).The T/M ratios of 99Tcm-HYNIC-ASONM group and control group were not significantly different at 1,4 and 10 h (t=2.154,2.287 and 2.236,all P＞0.05).Conclusion The antisense probe of MDM2 can accumulate specifically in prostate cancer tissue in animal models,which might be useful as a non-invasive genetic tool for the early diagnosis of prostate cancer.

Objective To investigate the effect of mouse double minute 2 (MDM2) mRNA ASON and mismatched oligonucleotide (ASONM) radiolabeled with 99Tcm on target gene expression in LNCaP cells.Methods The ASON and ASONM targeted to MDM2 mRNA were synthesized and radiolabeled by 99Tcm with the bifunctional chelator of HYNIC.The labeling efficiency,radiochemical purity,stability and molecular hybridization activity were investigated.The different concentrations of 99Tcm-HYNIC-ASON (0,100,500 nmol/L) and 99Tcm-HYNIC-ASONM (500 nmol/L) coated with lipofectamin 2000 were incubated with prostate cancer cells for 24 h,then RT-PCR and Western blot were carried out to assay the MDM2,p53 mRNA and the corresponding protein level.The variables of RT-PCR and Western blot were analyzed using one-way analysis of variance and q test.Results The labeling efficiency of ASON and ASONM were (65.15± 2.05)％ (n=5) and (64.93±2.18)％ (n=5),respectively.The radiochemical purity were both more than 90％.99Tcm-HYNIC-ASON had a good stability and could hybridize to the sense oligonucleotide (SON).The contents of MDM2 mRNA in 0,100,500 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups were 0.458±0.035,0.250±0.026,0.174±0.032,0.463±0.033,respectively,and there were significant differences between each 2 groups except between 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups (F=33.69,q =24.32-91.45,all P＜0.01).The average density of MDM2 protein in the 4 groups were 90.712±3.042,71.218±2.915,32.775±3.062,88.121±2.710,respectively (F=235.93,q=6.43-19.14,all P＜0.01; except 0 nmol/L99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).The contents of p53 mRNA in the 4 groups were 0.185±0.046,0.203±0.040,0.213±0.027,0.163±0.049,respectively(F =2.18,P＞ 0.05).The average density of p53 protein was 33.865 ± 2.213,70.445±2.180,99.025±3.012,38.351±3.271,respectively (F=53.98,q =3.32-6.74,all P＜0.01 ; except 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).Conclusions The MDM2 antisense probe can accumulate in the prostate cancer cells,and specially hybridize to the MDM2 mRNA and inhibit target gene expression.This novel molecular probe has a promising potential for the diagnosis of prostate cancer at gene level.

Objective To clone cDNA of trichosanthin (TCS) and purify TCS, and to study its influence on apoptosis and growth inhibition of colorectal carcinoma LoVo cells in vitro. Methods MTT assay was adopted to measure the growth inhibition ratio of LoVo cells treated with TCS, and apoptosis was assayed by agarose gel eletrophoresis. Results The results showed that the higher concentration of TCS and the longer testing time, the stronger growth inhibition of LoVo cells. DNA agarose gel eletrophoresis showed a gradient, which confirmed that TCS could induce the apoptosis of LoVo cells. Conclusions TCS can inhibit the growth of LoVo cells in vitro and induce its apoptosis. Our study provides evidence for the application of TCS in clinical treatment of human colorectal carcinoma.

Objective To formulate job evaluation index system for clinical nurse specialist in comprehensive children′s hospital. Methods A total of 19 experts were consulted by two-round Delphi method to ensure the item of index system, the order and weight of each index was determined by hierarchy analysis. Results The evaluation index system included 5 level-1 dimensions, 13 level-2 dimensions and 30 level-3 items. The authority of the expert advisory coefficient was 0.84, determine coefficient was 0.91, familiarity cofficient was 0.77. The coordination coefficient of one, two, three indexes were 0.44, 0.52, 0.51, with statistical significance (P < 0. 05). Conclusions The job evaluation index system is reliable and valid, which can provide basis on job management.

Objective To observe the effects of Huoxue Chubi Decoction on contents of PDGF-A and TGF-βin skin lesion of scleroderma mimicking mice models;To discuss its possible mechanism. Methods Thirty mice were randomly assigned into blank group, model group, TCM high-, medium-, low-dose group, with six mice in each group. Other than blank group, the rest of the 4 groups were given bleomycin-hydrocholoride 0.1 mL intradermal injection daily for 3 weeks to establish scleroderma mice models. From week 4 to 7, blank group and model group received gavage with equal amount of distilled water daily;TCM high-, medium-, low dosage group received gavage with Huoxue Chubi Decoction with different concentrations of 44.8, 22.4, 11.2 g/kg daily. The pathological changes in the skin tissue samples taken from each group were observed under microscope;the thickness of skin from each group was measured and the expression of PDGF-A, TGF-β, type-Ⅰ collagen (COL-Ⅰ) and type-Ⅲ collagen (COL-Ⅲ) were tested through immunohistochemical staining. Results Compare with blank group, the dermis tissue of model group was thicker, with the presence of thicker and greater number of collagen fibers, as well as infiltration of inflammatory cells. Compare with model group, thickness of skin and dermis collagen growth of TCM high-, medium-, low-dose group were milder varied by the concentration of the decoction. The result of immunohistochemical staining showed that the expressions of PDGF-A, TGF-β, COL-Ⅰ and COL-Ⅲ of modelgroup significantly increased than blank group (P<0.01); the expression of the mentioned indicators were statistically significant lower in the TCM high-, medium-, low-dose group than model group; The expressions of PDGF-A and TGF-β showed a positive correlation with the amount of COL-Ⅰ and COL-Ⅲpresented in each group. Conclusion Huoxue Chubi Decoction can suppress collagen expression and fibroblast formation in skin lesion of scleroderma mice models by down regulating the expressions of TGF-β and PDGF-A, thus demonstrate its potential in scleroderma treatment.

Objectives To investigate the effects of different degrees of carotid artery stenosis on cognitive function and neuronal apoptosis of hippocampal CA1 region in rats and to analyze the possible mechanisms of cognitive impairment. Methods According to the random number table,50 male Wistar rats were allocated into a sham operation group,a unilateral mild stenosis group,a unilateral moderate stenosis group,a unilateral severe stenosis group,a bilateral mild stenosis group,a bilateral moderate stenosis group,
a bilateral severe stenosis group,and a sham operation group. Aneedle-controlled suture method was used to induce a carotid stenosis model with different degrees of stenosis in rats. Water maze was to localize navigation and spatial search test was used to evaluate the cognitive function with different degrees of carotid artery stenosis in rats. Immunohistochemical method was used to observe the numbers of positive cells of P75 neurotrophin receptor (p75NTR),Bax,Bcl-2,neurotrophic factor-3 (NT-3 ),nerve growth factor (NGF)in hippocampal CA1 region under the light microscope. The conditions of neuronal apoptosis were observed. Results In 50 rats,6 died,1 rat in poor health failed to complete the Morris water maze test. The degree of bilateral carotid artery stenosis in 1 rat failed to meet 30%-60% at the same time,they were all removed. The remaining 42 rats were 6 in each group. (1)Compared with the sham operation group,the mean escape latency was prolonged (39 ± 6 s,32 ± 5 s,69 ± 7 s,respectively vs. 23 ± 4 s;all P < 0. 01),and the percentage of swimming distance in the platform quadrant was decreased (35 ± 4%,44 ± 4%,22 ± 5%,respectively vs. 53 ± 7%;all P < 0. 01),and the cognitive function was decreased with the degree of stenosis in the unilateral severe stenosis group,bilateral moderate stenosis group,and bilateral severe stenosis group. Compared with the unilateral severe stenosis group,the mean escape latency was prolonged and the percentage of swimming distance in the platform quadrant was decreased in the bilateral severe stenosis group (P < 0. 01). (2)Compared with the numbers of positive cells of Bax,p75NTR and Bcl-2 (8. 8 ± 3. 1,4. 2 ± 2. 3,and 5. 8 ± 1. 8,respectively)in the sham operation group,the numbers of positive cells of Bax,p75NTR,and Bcl-2 were increased (25. 5 ± 3. 5,11. 0 ± 2. 2,12. 3 ± 2. 7;15. 8 ± 3. 7,8. 9 ± 2. 2, 10. 5 ± 2. 9;and 47. 9 ± 6. 3,24. 7 ± 3. 0,12. 8 ± 2. 5,respectively)in the unilateral severe stenosis group, bilateral moderate stenosis group,and bilateral severe stenosis group (all P < 0. 01). The numbers of Bax and p75NTR positive cells were increased with the degree of stenosis. When the stenosis was severe,the numbers of Bax and p75NTR positive cells were increased in the bilateral severe stenosis group compared with those of the unilateral severe stenosis group (P < 0. 01). The numbers of NGF and NT-3 positive cells in each stenosis group had an increased trend compared with sham operation group,but there were no significant differences (F =1. 034,and 1. 358;P = 0. 420 and 0. 259 respectively). Conclusions Carotid stenosis can cause cognitive disorder in rats,and it is correlated with the degree of carotid stenosis. Ischemia caused neuronal apoptosis in hippocampal CA1 region may be one of the mechanisms of cognitive impairment after carotid artery stenosis in rats.

To observe the clinical therapeutic effect of diabetic peripheral neuropathy (DPN) treated by needling combined with drug, 104 patients with DPN were randomly divided into acupuncture plus drug group and control group, and each group had 52 patients. After treatment of two months, the clinical effective rate in acupuncture plus drug group was 51.9%, and the total effective rate was 88.5%, both of them were better than those in control group (P＜0.05). The needling method of nourishing the kidney and dredging the meridian combined with drug had good clinic effect in the treatment of DPN.

Objective To investigate the expression and the significance of toll-like receptor 3 (TLR-3)in placenta,tumor necrosis factor-α(TNF-α)in maternal and cord blood of idiopathic fetal growth restriction(IFGR),and their correlation with the pathogenesis of symmetric and asymmetric IFGR.Methods From April 2008 to April 2009,42 primiparae of singleton pregnancy and their IFGR babies,who delivered at term through cesarean section, in the Third Affiliated Hospital of Zhengzhou University were enrolled. All subjectects were divided into symmetric IFGR group (n=20) and asymmetric IFGR group (n =22). Another 42 non-IFGR pairs were randomly selected as the control group. The polink-2 plus polymerized horseradish peroxidase (HRP) immunohistochemical method and the enzyme linked immunosorbent assay (ELISA) were applied to detect TLR-3 and TNF-α levels. Results (1) The expression of TLR-3 protein were observed in all maternal placenta of the three groups. TLR-3 essentially expressed in syncytiotrophoblasts and hofbouer cells in the symmetric IFGR and control group, but expressed mostly in hofbouer cells and less in syneytiotrophoblasts in the asymmetric IFGR group. (2) The expression of TLR-3 in the syncytiotrophoblasts of the symmetric and asymmetric IFGR group was significantly lower than in the control group (111±14 and 118±11 vs. 156 ± 9, P<0. 01). The number of TLR-3 positive in Hofbourer cell in the symmetric IFGR group was lower than the control group (8. 9±2. 8 vs 17.5±2. 8, P <0. 01 ), but the number in the asymmetric IFGR group was higher (23.8±3.7) compared with the control group (P <0. 01). (3) The TNF-α levels in the maternal and cord blood of the symmetric and the asymmetric group were higher than that of the control group [maternal : (90±10) μg/L and ( 86±11 ) μg/L vs. (73±9) μg/L;cord blood: (92±12) μg/L and (96±8) μg/L vs. (79±9) μg/L;P<0.01]. (4) Neither symmetric nor the asymmetric IFGR group showed any correlations between the maternal and cord blood levels of TNF-α (P>0. 05). (5) Significant correlation was found between the TNF-α level of the cord blood and TLR-3 expression in the placenta in both the symmetric and asymmetric IFGR group(P<0. 05),but no relationship was found between the maternal blood TNF-α level and TLR-3 expression in the placenta (P>0. 05). Conclusions The variantions of TLR-3 expression in placenta and the increased expression of TNF-α in cord blood are associated with the genesis IFGR. The reduced expression of TLR-3 may related to symmetric IFGR, while the increased TLR-3 level in hofbouer cells may lead to asymmetric IFGR.