Effects of Pb,Zn,Cu and Cd on growth and enzyme activities of microorganisms such as Bacillus licheniformis 2709,Bacillus subtilis1.398 and Aspergillus usamii 537,which can individually produce alka-line,neutral and acid protease,were studied,respectively. By determining growth and enzyme activities of three bacterial species,as well as analyzing their tolerance and resistance indicators(MTC and MIC) using growth inhibition plate analysis(GIPA) under different heavy metal gradients,results showed that four heavy metals under higher than a special concentration level significantly inhibited growth and enzyme ac-tivities of three species. The alkaline protease exhibited greatest adaptability to four heavy metals,next the neutral protease also exhibited certain adaptibilty to Zn and Cd,whereas the acid protease was totally inhib-ited under any levels of heavy metal concentrations. Three species not only showed great potential to tolerate and resist toxicity of lead and zinc(2.0 mmol/L-6.0 mmol/L) ,but also showed certain resistance to cad-mium(0.5 mmol/L-0.75 mmol/L) .

Objective To evaluate the clinic value of multiple tumor markers plus PSA in diagnosing prostate cancer.Methods We collected serum samples of 140 prostate cancer patients with average age of 68 (48-82) years,104 benign prostate hyperplasia patients with average age of 70 (52-87) years,and 162 healthy people with average age of 38 (23-49) years.We had detected PSA levels and also the protein expressions of XAGE-1b,SSX-2,AM ACR and AKAP4.In healthy people,the ranges of the normal values with 95％ data range were determined.Multiple tumor markers and PSA were calculated their positive rate,specificity and sensitivity in diagnosing prostate cancer.We had randomly chosen one positive serum,and analyzed the 4 protein expressions by Western bolt.We detected the serum with the four markers plus PSA by Luminex medthod,then drew ROC curve and calculated AUC area according to the results.Results Among all those samples,PSA levels of 266 samples were under 4tμg/L,86 samples were between 4-10 μg/L,and 54 samples were above 10 μg/L.The positive rates of XAGE-1b,SSX-2,AMACR and AKAP4 in prostate cancer patients were 53.6％ (75/140),34.3％ (48/140),27.9％ (39/140),44.3％ (62/140) respectively.The AUC of XAGE-1b,SSX-2,AMACR,AKAP4 and PSA were 0.666,0.615,0.551,0.768,0.675 respectively.The AUC of their combination was 0.887; The specificity and sensitivity of single PSA detection and combination detection were 60.0％,46.2％ and 80.0％,82.2％.Among those whose PSA value was between 4-10 μg/L,the AUC of single PSA detection was 0.505,the specificity and sensitivity of single PSA detection were 43.2％ and 31.8％; the AUC of the combination detection was 0.803; their specificity and sensitivity were 83.7％ and 73.2％.Conclusions Compared with the single PSA detection,the combination of XAGE-1b,SSX-2,AMACR,AKAP4 and PSA has been greatly improved the specificity and sensitivity in prostate cancer detection.This tool still has significant value even in patients with PSA value between 4-10 μg/L.

OBJECTIVE:To study the influential factors related to efficacy of Peach kernel-Rheum palmatum couplet medi-cines in TCM formula,and to reveal the general regularity of compatibility environment,common ratio,processing variety and dosage forms of P. kernel-R. palmatum couplet medicines. METHODS:Using Chinese Medical Prescription Selected Dictionary ed-ited by Peng Huairen as data source,142 formulas of P. kernel-R. palmatum couplet medicines were collected. By establishing data-base,compatibility types of P. kernel-R. palmatum couplet medicines,as well as common ratio,processed prodact,dosage form were classified statistically. The influential factors related to efficacy of P. kernel-R. palmatum couplet medicines with different pro-portions were summarized. RESULTS:The efficacy of P. kernel-R. palmatum couplet medicines could be divided into 6 aspects and 11 roles,including activating blood circulation to dissipate blood stasis(activating blood to relieve pain,promoting blood circula-tion to eliminate disease,activating blood to promote menstruation,breaking stagnant and eliminating blood stasis),eliminating carbuncle and detoxicating(cleaning intestine and clearing away the pathogenic heat of lung,eliminating carbuncle and expelling pus,eliminating sore and detoxicating),expelling the pathogenic heat to loosen the bowels,warming yang for dispelling cold,forti-fying the spleen and nourishing the stomach,relaxing tendon and activating blood. The compatibility environment of P. kernel-R. pal-matum couplet medicines were mainly compatible with TCM for activating qi to eliminate stasis,activating blood to promote menstru-ation,breaking stagnant and eliminating blood stasis,expelling the pathogenic heat to expel stasis. The ratio of P. kernel to R. palma-tum ranged 1 : 8-4 : 1,and the ratio ranged 1 : 8-3 : 1 when performing the role of actirating blood circalation to dissipate blood stasis. Common processed products were crude P. kernel and prepared R. palmatum. Common dosage forms were mainly decoction,pill and powder. CONCLUSIONS:Compatibility environment,ratio,processing varieties,dosage forms influence the effects of P. kernel-R. palmatum couplet medicines,especially compatibility environment.

OBJECTIVE:To establish a method for the contents of gallic acid,catechin,sennosides B,aloe-emodin,rhein, emodin,chrysophanol,physcion,chrysophanol-1-O- glucoside and emodin-8-O- glucoside in Rhei Radix et Rhizoma,Jiu Rhei Radix et Rhizoma,Shu Rhei Radix et Rhizoma,Rhei Radix et Rhizoma tan,Cu Rhei Radix et Rhizoma,and analyze the differ-ences. METHODS:HPLC was performed on the column was Hypersil C18 with mobile phase of methanol- 0.2% acetic acid(gradi-ent elution)at a flow rate of 1.0 ml/min,the detection wavelength was 260 nm,column temperature was 25 ℃,injection volume was 10 μl. RESULTS:The linear range was 0.252 5-4.040 0 μg for gallic acid(r=0.999 6),0.600 0-9.600 0 μg for catechin(r=0.999 6),0.297 4-4.758 4 μg for sennosides B(r=0.999 9),0.001 8-0.028 8 μg for aloe-emodin(r=0.999 9),0.005 0-0.080 0 μg for rhein(r=0.999 9),0.019 0-0.304 0μg for emodin(r=0.999 8),0.380 2-6.083 2μg for chrysophanol(r=0.999 7),0.008 2-0.131 2μg for physcion(r=0.999 8),0.126 0-2.016 0 μg for chrysophanol-1-O-glucoside(r=0.999 6)and 0.111 3-1.780 8 μg for emo-din-8-O-glucoside (r=0.999 8);RSDs of precision,stability and reproducibility tests were lower than 3.0%;recoveries were 96.17%-97.21%(RSD=1.67%,n=6),97.60%-100.54%(RSD=2.55%,n=6),99.45%-101.32%(RSD=1.63%,n=6), 95.31%-98.19%(RSD=2.42%,n=6),98.99%-100.35%(RSD=1.86%,n=6),98.95%-101.21%(RSD=2.17%,n=6), 99.81%-100.62%(RSD=1.66%,n=6),96.78%-98.52%(RSD=1.99%,n=6),97.80%-100.14%(RSD=3.32%,n=6) and 97.40%-101.24%(RSD=2.89%,n=6). Compared with Sheng Rhei Radix et Rhizoma,the contents of gallic acid,catechin,sen-nosides B and anthraquinones in Cu Rhei Radix et Rhizoma,Jiu Rhei Radix et Rhizoma and Rhei Radix et Rhizoma tan decreased. The contents of catechin,sennosides B and anthraquinones in Shu Rhei Radix et Rhizoma. Catechin,sennosides B,chrysopha-nol-1-O- glucoside,aloe-emodin and rhein were not detected in Dahuang tan. CONCLUSIONS:The method is simple with good precision,stability and reroducibility,and can be used for the simultaneous determination of 10 chemical components in processed products of Rhei Radix et Rhizoma;there were significant differences in contents of 10 chemical components in processed prod-ucts of Rhei Radix et Rhizoma.

AIM: To investigate the biocolonization of poly 2-hydroxyethy methacrylate (PHEMA) sponge with cornea tissue and evaluate the therapeutic effects of modified porous poly 2-hydroxyethy methacrylate-Polymethyl met-hacrylate (PHEMA-PMMA) Keratoprostheses (KPro) on rabbit and monkey corneas. METHODS:The KPro were made using two-stage polymerization combined with mechanical cutting. The experiment was divided into two groups. In the control group, ten normal rabbit eyes received lamellar implanta-tion of PHEMA sponges. The sponges were obtained 2 weeks, 1,2,3 and 4 months after operation. The cell proliferation and neovascularization inside the sponges were observed using light and transmission electron microscopy (TEM) and immunohistochemistry. In the experimental group, the porous PHEMA-PMMA KPros were inserted into the lamellar pockets of eight rabbit corneas and two monkey corneas (stage I operation). The healing process was investigated by slit-lamp microscopy. The anterior lamellar cornea tissues were removed 3 months after surgery, exposing the under-neath transparent core (stage II operation). The operated eyes were then followed up for 3-6 months.light microscope, fibroblasts started to grow into the cornea 2 weeks after operation; lots of cells, accompanied with new blood vessels, invaded into the cornea 2-3 months after surgery. Invading cells of sponge, as well as keratocytes, were positive for vimentin. Under the electron microscope, the invading cells looked healthy and were surrounded by extracellular matrix and collagen. In 8 rabbit eyes which received KPro implantation, anterior lamellar cornea melting happened in two eyes after the stage II operation. The remaining 6 corneas retained their central cores during observation after the stage II operation.Two monkey operated eyes were found no complication thoughout the whole follow-up.cornea. The modified PHEMA-PMMA KPros have obtained a relatively stable results after implantation into animal corneas.

Objective To compare the sensitivity and specificity of dot immunogold method (DIM) and particle agglutination (PA) for the diagnosis of mycoplasma pneumoniae (MP) infection. Methods The 190 serum specimens of 113 children with mycoplasmal pneumonia (infection group) and 50 serum specimens of 50 health children (health group) were tested for MP by PA and DIM- A and B. Results In infection group, the positive rates of DIM- A and B were 82.63% (157/190) and 84.74%(161/190), and there was no statistical difference (χ2 = 0.31, P>0.05); the positive rate of PA (titer ≥1:160) was 70.00%(133/190), the positive rate of PA was significantly lower than that in DIM-A and B, and there were statistical differences (P<0.05). In infection group, with the increase of PA titer, the positive rate of DIM was gradually increased, and there was a correlation between 2 methods (rA=0.972, rB=0.830);the positive rates of DIM- A and B in serum specimens of PA negative were 40.62%(13/32) and 53.12%(17/32), and in the PA titer of 1:5120, there was still a negative result in DIM. In health group, the positive rates of DIM-A and B were 26.00% (13/50) and 28.00% (14/50), and there was no statistical difference (χ2 = 0.66, P>0.05); the positive of PA was 8.00% (4/50), the positive rate of PA was significantly lower than that in DIM- A and B, and there were statistical differences (P<0.05 or<0.01). Conclusions Compared with the PA, DIM has low sensitivity and poor specificity for clinical diagnosis. DIM is not suitable for clinical diagnosis of MP infection.

Equipment Design ; Facial Injuries ; therapy ; First Aid ; instrumentation

The fruit of Pandanus tectorius (PTF) has a long history of use as a folk medicine to treat hyperlipidemia in Hainan province, South China. Our previous studies have shown that the n-butanol extract of PTF is rich in caffeoylquinic acids and has an adequate therapeutic effect on dyslipidemic animals induced by high-fat diet. In this work, seven caffeoylquinic acids isolated from PTF were screened for the lipid-lowering activity in HepG2 hepatoma cells. Oil-Red O staining, microscopy and intracellular triglyceride (TG) and total cholesterol (TC) quantification showed that 3-O-caffeoylquinic acid (3-CQA), 3, 5-di-O-caffeoylquinic acid (3,5-CQA), and 3,4,5-tri-O-caffeoylquinic acid (3,4,5-CQA) significantly inhibited lipid accumulation induced by oleic acid and decreased intracellular levels of TC and TG in a dose-dependent manner. These three caffeoylquinic acids showed no significant cytotoxicity at concentrations of 1 -50 μmol x L(-1) as determined by MTT assay. Realtime quantitative PCR revealed that 3-CQA and 3, 5-CQA significantly increased the expression of lipid oxidation-related genes PPARα, CPT-1 and ACOX1 while 3-CQA, 3, 5-CQA and 3,4,5-CQA decreased the expression of lipogenic genes SREBP-1c, SREBP-2, HMGR, ACC, FAS. Overall, 3-CQA, 3, 5-CQA and 3, 4, 5-CQA may be the principal hypolipidemic components in PTF which can decrease intracellular lipid accumulation through up-regulating the expression of lipid oxidative genes and down-regulating the expression of lipogenic genes.
Carcinoma, Hepatocellular ; metabolism ; China ; Cholesterol ; metabolism ; Gene Expression Regulation ; Hep G2 Cells ; Humans ; Lipid Metabolism ; Liver Neoplasms ; metabolism ; Oleic Acid ; Pandanaceae ; chemistry ; Quinic Acid ; analogs & derivatives ; chemistry ; Sterol Regulatory Element Binding Protein 1 ; Triglycerides ; metabolism

Objective To assess the applicability of the Modification of Diet in Renal Disease (MDRD)formula to kidney function impaired Chinese diabetic patients.Methods Glomerular filtration rates(GFRs)in 463 Chinese diabetic patients(219 female,244 male,aged 14 to 88)were estimated by measuring ~(99m)Tc-DTPA clearance and with equations based on serum creatinine(Scr)and cystatin C(Cys C)concentrations.GFRs derived from various equations were compared with the ~(99m)Tc-DTPA clearance GFRs and their relative accuracies were assessed with ROC analysis.All the Scr measurements were performed with both the Roche enzymatic assay and the Beckman LX20 kinetic alkaline picrate assay,and Cys C with immunonephelometric and immunoturbidimetric assays.Results The reciprocals of Cys C and Scr were linearly correlated with ~(99m)Tc-DTPA clearance GFRs(r=0.830 and 0.690,repectively).The correlation of GFR with Scr could be expressed by an adjusted MDRD equation:GFR [ ml?(min?1.73 m~2)~(-1)]=175?(Scr)~(-1.154)?(age)~(-0.203)?0.742(female)?0.827,where 0.827 was a coefficient for Chinese.The adjusted equation showed a better accuracy than the MDRD equation(areas under the ROC curve 0.818 vs 0.644).The adjusted equation was also more accurate than equations obtained in previous Chinese studies.GFRs were also estimated by using Cys(in mg/L)with the following equation:GFR [ ml?(min?1.73 m~2)~(-1)] = 63.24?(Cys C)~(-0.3378).The accuracy of the Cys equation was similar to the Scr equation,or better in patients aged 60 and above.The Roche enzymatic results which were traceable to the isotope dilution mass spectrometry(IDMS)methods were significantly lower than Beckman LX20 results,but the results were closely correlated with each other(Y = 0.94X-0.02).When non-traceable Scr results were used,the coefficient needed to be adjusted.Conclusions GFRs can be estimated with equations based on either Scr or Cys C.GFR estimation should use standardized Scr results and take into account ethnic effects.

Objective To identify seven species of Gynostemma BI.,including G.pentaphyllum,G. pentagynum,G.cardiospermum,G.longipes,G.yixingense,G.laxiflorum,and G.guangxiense,by in- ter-simple sequence repeat(ISSR)markers.Methods General DNA was isolated from leaves of the seven species in Gynostemma B1.by CTAB,57 primers constituted by ISSR were tested for PCR and sepharose electrophoresis.Results Fourteen primers amplified polymorphic bands,the amplification patterns of primers UBC-873 and UBC-895 were higher in terms of polymorphic and amplified band ratio.They are used to distinguish all the examined seven species.Conclusion ISSR-PCR Method provides a quick,reli- able molecular marker technique for the identification of different species of Gynostemma B1.