Objective:To explore the correlation among serum cardiac troponin I (cTnI),N terminal pro brain natri-uretic peptide (NT-proBNP)and main endpoint events of heart in patients with chronic stable heart failure.Meth-ods:The present study enrolled 95 patients with NYHA cardiac function class III~IV from Feb 2010 to Feb 2011.According to levels of cTnI and NT-proBNP,the patients were divided into cTnI negative group (n=60)and cTnI positive group (n=35);NT-proBNP negative group (n=40)and NT-proBNP positive group (n=55),all patients were followed up for two years,and the main endpoint events were cardiogenic sudden death and rehospitalization caused by acute aggravation of heart failure.Results:Compared with negative group,the hazard ratio (HR)of end-point events was 2.69 and confidence interval (CI)was 1.54~ 4.72,P = 0.002 in cTnI positive group;HR was 2.54 and CI was 1.35~4.78,P =0.003 in NT-proBNP positive group;further interclass crossover analysis found that,when patients'cTnI and NT-proBNP were both positive,the hazard ratio of cardiac endpoint events was the highest (HR=6.34,CI 2.26~17.9,P <0.001).Conclusion:In patients with chronic stable heart failure,serum elevated levels of cardiac troponin I and N terminal pro brain natriuretic peptide are important predictors reflecting prognosis of patients with heart failure.

Objective To get the experience of surgical treatment for diabetic foot (DF) ulcer.Methods Clinical data of 85 patiems (108 limbs in total) admitted in Department of Bum and Plastic Surgery in The First Affiliated Hospital of Chongqing Medical University from Jan 2010 to Dec 2012 were retrospectively analyzed.Results Wound culture results of the 85 patients were:bacteria in 142 limbs,and fungi in 7 limbs.The main bacteria cultured were staphylococcus aureus,escherichia coli,coagulase negative staphylococcus,enterococcus,pseudomonas aeruginosa,and acinetobacter baumannii.DF healing and Wagner classification was negatively correlated:the higher the class,the less the primary healing rate.The higher amputation rate were found in the higher class patients.The more delayed healing,and the longer the healing time and average hospital stay were also found in the higher class patients.85 patients underwent surgery:skin grafting in 45 cases (52.9％),flap in 8 cases (9.4％),skin repair + flap grafting in 7 cases,toe (limb) amputation in 28 cases,2 cases were sutured after debridement.All cases were cured after one or multiple surgeries.No recurrence was found during the follow-up of 10 days to 2 months.No delayed infection occurred to the repairments.The shape and weight bearing walking function were good.Conclusions The treatment of DF need to follow the principle of multidisciplinary cooperation,professional treatmem,systemic and local treatment.We need to pay attention to the etiology and prevention of the disease,focus on wound bed preparation,choose the best treatment.Early operation can significandy fascilitate wound healing.

OBJECTIVE:To establish a method for the content determination of berberine hydrochloride in the Jiangtang xiao-zhi tablets. METHODS:HPLC was conducted with the Symmetry C18 column. The mobile phase was acetonitrile-0.05 mol/L sodium dihydrogen phosphate(pH adjusted to 3.0 using phosphoric acid)(24∶76,V/V)at the flow rate of 1.0 ml/min. The detection wave-length was 345 nm,the column temperature was room temperature and injection volume was 10 μl. RESULTS:The linear range of berberine hydrochloride was 0.522-4.698 μg(r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 0.72%;the average recovery was 97.79%(RSD=2.09%,n=6). CONCLUSIONS:The method is simple,accurate,sensitive and reproducible,and can be used for the content determination of berberine hydrochloride.

Aim To investigate the protective effect of astragaloside IV (AS-Ⅳ) on human retinal pigment epithelium injury induced by methylglyoxal (MGO), and explore its molecular mechanism.Methods The injury of ARPE-19 cells was induced by MGO and the cell viability was measured by CCK-8 method.The morphology of cell nucleus was analyzed by Hoechst 33342 staining and the cell apoptosis was analyzed by flow cytometry to detect labbled Annexin V-FITC/PI.JC-1 staining and fluorescence probe DCFH-DA were employed to evaluate the change of mitochondrial membrane potential and reactive oxygen species (ROS).The levels of SOD, MDA, caspase-9 and caspase-3 were determined by respective kits.Western blot was used to analyse the expression of Bcl-2, Bax and PARP.Results AS-Ⅳ could significantly inhibit the decrease of cell viability induced by MGO, improve the morphology of cell nucleus, reduce the ARPE-19 cell apoptosis rate and the level of ROS and MDA, and increase the activity of SOD.Furthermore, AS-Ⅳ could enhance mitochondrial membrane potential, the ratio of Bcl-2/Bax and the expression of PARP, and inhibit the activation of caspase-9 and caspase-3.Conclusion AS-Ⅳ may protect ARPE-19 cells from the injury induced by MGO by increasing the antioxidant ability of ARPE-19 cells and inhibiting cell apoptosis.

Aim To investigate the optimum trypsin digestion time and the optimum trypsin concentration of the long-term formalin-fixed retina in rats.Methods Male SD rats were sacrificed by cervical dislocation. The removed eyes were fixed in 4% formaldehyde fixa-tive for 2 days,2 weeks,4 weeks,3 months,5 months and 7 months respectively.Fixed retinas at dif-ferent time were separated and put in 3%,6% and 9% trypsin at 37℃ in incubator.The time required was recorded for complete digestion of retinas,which fixed for different time and digested in different con-centrations of trypsin.The retinal blood vessels were recorded under a microscope after PAS staining of stretched retinal blood vessel.Results After conven-tionally fixed for 48 hours,retinas were digested com-pletely in 3% trypsin at 37℃ for 3.5 hours.The di-gestion time extended to 9 hours and 1 4 hours for the 2 weeks and 1 month fixed eyes,using 3% trypsin, which achieved the same result as conventional fixed eyes.For the 3 months fixed eyes,a complete retinal blood vessel could also be attained by using conven-tional concentrations of trypsin,and only by extending digestion time to 22 hours.It is also indicated that the increased concentration of trypsin did not shorten the time of digestion.In addition,for the fixed eye from 5 months to 7 months,6% and 9% trypsin could short-en,to some extent,the time to 22 hours and 28 hours respectively,which was conductive to obtaining rela-tively complete retinal blood vessels.Conclusions Retinal tissues which fixed time shorter than 3 months can completely be digested using 3% trypsin.Howev-er,for retinal tissue,which fixed from 5 months to 7 months,both the extending time of digestion and the increasing concentration of trypsin to 6% are necessary to get optimum retinal vascular digestion.

Objective Recombinant human parathyroid hormone(1-34) [ rhPTH(1-34)] is the unique anabolic substance acting on the skeleton. The efficacy and safety of long-term administration of rhPTH(1-34) in Chinese postmenopausal women have not been evaluated. This study compared the clinical efficacy and safety of rhPTH(1-34) with elcatonin for treating postmenopausal women with osteoporosis in 11 urban areas of China. Methods A total of453 postmenopausal women with osteoporosis were enrolled in an 18-month, multi-center, randomized, controlled study. They were randomized to receive either rhPTH(1-34) 20 μg(200 U) daily for 18 months, or elcatonin 20 U weekly for 12 months. Lumbar spine ( L1-4) and femoral neck bone mineral density (BMD), fracture rate, back pain as well as biochemical markers of bone turnover ( serum bone-specific alkaline phosphatase was measured by radioimmunoassay; C-telopeptide/ creatinine ( CTX/ Cr) measured by quantitative sandwich enzyme-linked immunosorbent assay) at 6, 12, and 18 months. Adverse events were recorded. Results rhPTH(1-34) increased lumbar BMD more significantly than that did by elcatonin at 6 months( M6), 12 months (M12), and 18 months(M18; 4. 3% vs 1. 94% , 6. 8% vs 2. 72% , 9. 51% vs 2. 86% , P<0. 01). There was only a small but significant increase of femoral neck BMD at M18(2. 64% , P<0. 01) in rhPTH(1-34) groups. There were greater increases in bone turnover markers in the rhPTH(1-34) group than in the elcatonin group at M6, M12, and M18[serum bone-specific alkaline phosphatase(BSAP) 93. 67% vs -3. 56% , 117. 78% vs -4. 12% , 49. 24% vs-5. 81% , P<0. 01; urinary CTX/ Cr 250% vs -29. 5% , 330% vs -41. 4% , 273 % vs -10. 6% , P<0. 01]. rhPTH (1-34) showed similar effect of pain relief as elcatonin. The incidence of clinical fractures was 5. 36% (6 / 112) in elcatonin group and 3. 23% ( 11 / 341 ) in rhPTH ( 1-34 ) group ( P = 0. 303 ). Both treatments were well tolerated. Hypercaluria(9. 38% ) and hypercalcemia(7. 04% ) in rhPTH(1-34) group was transient and caused no clinical symptoms. Pruritus(8. 21% vs 2. 68, P=0. 044) and redness of injection site(4. 40% vs 0, P=0. 024) were more frequent in rhPTH(1-34). Nausea / vomiting(16. 07% vs 6. 16% , P = 0. 001) and hot flushes(7. 14% vs 0. 59% , P<0. 001) were more common in elcatonin group. Conclusion rhPTH(1-34) treatment was associated with greater increases in lumbar spine BMD and bone formation markers. It could increase femoral BMD after 18 months treatment. rhPTH(1-34) could ameliorate back pain effectively. The results of the present study indicate that rhPTH(1-34) is an effective, and safe agent in treating postmenopausal women with osteoporosis.

OBJECTIVE To study the toxicity of the extract from Mi ao medicine Wikstroemia indica to zebrafish . METHODS Zebrafish embryo model was used as the object ，after exposure to W. indica extract （10，20，40 μg/mL），the number of spontaneous twitching within 1 min，heart rate within 10 s，the occurrence of malformation and death were detected and recorded . Zebrafish was used as the object ，after exposure to W. indica extract（10-100 μg/mL）for 24，48 and 72 h，and the median lethal concentration（LC50）of W. indica extract to zebrafish at different time points were calculated . After exposure to low ，medium and high concentration （27，37，51 μg/mL）of W. indica extract，the liver phenotype ，hepatocyte apoptosis and lipid deposition of zebrafish were observed ，and the activities of alanine aminotransferase （ALT），aspartate aminotransferase （AST）and lactate dehydrogenase（LDH）in liver tissue were detected . RESULTS Compared with blank group ，the number of spontaneous twitching ， malformation rate （except for 10 μg/mL group ）and mortality of embryos increased significantly in 10，20，40 μg/mL groups of W. indica extract，and the heart rate （except for 10，20 μg/mL group ）of embryos decreased significantly （P＜0.05）. LC50 of W. indica extract to zebrafish at 24，48 and 72 h were 39.850，28.300 and 21.490 μg/mL，respectively. After drug treatment ，the transparency of liver area of zebrafish in low ，medium and high concentration groups of W. indica extract reduced and their shape were enlarged；apoptosis and lipid deposition increased ；the activities of ALT ，AST and LDH in liver tissue were significantly increased （P＜0.05）. CONCLUSIONS Miao medicine W. indica extract had develop mental toxicity to zebrafish embryos and he patotoxicity to zebrafish .

Natural cyclohexapeptide AFN A₁ fromStreptomyces alboflavus 313 has moderate antibacterial and antitumor activities. An artificial designed AFN A₁ homodimer, di-AFN A₁, is an antibiotic exhibiting 10 to 150 fold higher biological activities, compared with the monomer. Unfortunately, the yield of di-AFN A₁ is very low (0.09 ± 0.03 mg·L^-1) in the engineered strain Streptomyces alboflavus 313_hmtS (S. albo/313_hmtS), which is not friendly to be genetically engineered for titer improvement of di-AFN A₁ production. In this study, we constructed a biosynthetic gene cluster for di-AFN A₁ and increased its production through heterologous expression. During the collection of di-AFN A₁ biosynthetic genes, the afn genes were located at three sites of S. alboflavus 313 genome. The di-AFN A₁ biosynthetic gene cluster (BGC) was first assembled on one plasmid and introduced into the model strain Streptomyces lividans TK24, which produced di-AFN A₁ at a titer of 0.43 ± 0.01 mg·L^-1. To further increase the yield of di-AFN A₁, the di-AFN A₁ BGC was multiplied and split to mimic the natural afn biosynthetic genes, and the production of di-AFN A₁ increased to 0.62 ± 0.11 mg·L^-1 in S. lividans TK24 by the later strategy. Finally, different Streptomyces hosts were tested and the titer of di-AFN A₁ increased to 0.81 ± 0.17 mg·L^-1, about 8.0-fold higher than that in S. albo/313_hmtS. Successful heterologous expression of di-AFN A₁ with a remarkable increased titer will greatly facilitate the following synthetic biological study and drug development of this dimeric cyclohexapeptide.
Cloning, Molecular ; Streptomyces/metabolism* ; Multigene Family ; Anti-Bacterial Agents/metabolism* ; Plasmids/genetics*

l-Heptopyranoses are important components of bacterial polysaccharides and biological active secondary metabolites like septacidin (SEP), which represents a group of nucleoside antibiotics with antitumor, antifungal, and pain-relief activities. However, little is known about the formation mechanisms of those l-heptose moieties. In this study, we deciphered the biosynthetic pathway of the l,l-gluco-heptosamine moiety in SEPs by functional characterizing four genes and proposed that SepI initiates the process by oxidizing the 4'-hydroxyl of l-glycero-α-d-manno-heptose moiety of SEP-328 ( 2) to a keto group. Subsequently, SepJ (C5 epimerase) and SepA (C3 epimerase) shape the 4'-keto-l-heptopyranose moiety by sequential epimerization reactions. At the last step, an aminotransferase SepG installs the 4'-amino group of the l,l-gluco-heptosamine moiety to generate SEP-327 ( 3). An interesting phenomenon is that the SEP intermediates with 4'-keto-l-heptopyranose moieties exist as special bicyclic sugars with hemiacetal-hemiketal structures. Notably, l-pyranose is usually converted from d-pyranose by bifunctional C3/C5 epimerase. SepA is an unprecedented monofunctional l-pyranose C3 epimerase. Further in silico and experimental studies revealed that it represents an overlooked metal dependent-sugar epimerase family bearing vicinal oxygen chelate (VOC) architecture.