The nitrate-N content in KNO3 solution and soil was rapidly predicted using techniques of mid-infrared spectroscopy, in which 15 NO-3 and 14 NO-3 were distinguished and predicted. The results showed that the characteristic band of nitrate in solution and soil was 1200-1500 cm-1 , and compared with 14 NO-3 , the red shift of characteristic band of 15 NO-3 was about 35 cm-1 . In the characteristic band of nitrate, absorption band increased with the nitrate nitrogen concentration with less interference absorption. The linear regression was made between the first principal component of characteristic band and nitrate-N content, and correlation coefficient was more than 0 . 9840 , indicating that the technique of mid-infrared attenuated total reflectance spectroscopy could be applied for rapid monitoring of nitrate in solution and soil. Meanwhile, based on the red shift characteristic of 15 NO-3 absorption band, the method of partial least squares were involved to predict the nitrate-N of different N-isotope labeled in solution and soil, resulting that all the prediction models reached excellent levels. For 14 NO3-N and 15 NO3-N in solution, the correlation coefficients ( R2 ) were 0. 9980 and 0. 9982 respectively, and ration performance to standard deviations ( RPD ) were 6. 44 and 4. 76, respectively. While for 14 NO3-N and 15 NO3-N in soil, the correlation coefficients ( R2 ) were 0. 9794 and 0. 9679, and RPD were 5. 75 and 4. 78, respectively. Therefore, the technique of mid-infrared attenuated total reflectance spectroscopy can be applied for rapid monitoring different N-isotope labeled nitrate in solution and soil, to provide a new in situ and fast time method to study nitrification process in soil.

Objective To explore the feasibility of fluorescence quantitative PCR(FQ-PCR)in prenatal diagnosis of the fetal RhD genotyping using free DNA from RhD-negative pregnant women.Methods The fetal RhD gene was amplified from 78 RhD-negative pregnant women with single fetus maternal plasma (gestation from 11 to 40 weeks).Rhe existence of fetal DNA was confirmed by amplification ofnine different polymorphic short tandem repeat loci(STR)and sex-determining region Y chromosome(SRY)gene.Exon5,7,10 and intron 4 were amplified by real-time polymerase chain reaction with TaqMan probe.The results of fetal RhD genotyping were evaluated retrospectively by the serologic analysis of infant cord blood.Results Among the 78 specimens,the SRY positive signals were detected from samples of 41 and were all identified male fetal through 8ex observation after newborn infants delivered from the women enrolled.The mean concentration of SRY gene reached(214.7±120.9)eopies/ml.RhD genotyping results of 70 cases were in complete concordance with the resets through serological detection of fetal cord blood after delivery.In addition,5 cases were false-positive.3 cages were considered inconclusive.The coincidence rate was 90%(70/78).From 5 false-positive cases,4 cases were identified as RhDel phenotype by detecting RHDl227A allele gene.The final accuracy rate of FQ-PCR was 95%(74/78)in the fetal RhD genotyping.Conclusion FQ-PCR analysis for noninvasive prenatal of fetal RhD genotyping could be useful in prevention and diagnosis of hemolytic disease of newborn.

Objective To establish the methodology of uncertainty evaluation with reference measurement procedure by Monte Carlo method (MCM).Methods According to JCGM 101:2008,we established the methodology of uncertainty evaluation by MCM in the example of GGT reference measurement procedure.We could calculate an estimate of GGT concentration,the associated standard uncertainty and a coverage interval with a specified probability by MATLAB software,setting the uncertainty evaluation by GUM method as a control.Results When the uncertainty was evaluated by GUM method,the results of sample A and Sample B were (95.8 ±2.4) U/L (k =2) with coverage interval [93.4 U/L,98.2 U/L] and (180.0 ± 3.9) U/L (k =2) with coverage interval [176.1 U/L,183.9 U/L] respectively,while using MCM method,the uncertainty evaluation result of sample A and Sample B were (95.8 ± 2.4) U/L (k =2) with coverage interval [93.4 U/L,98.2 U/L] and (180.0 ± 3.9) U/L with symmetrical 95％ coverage interval [176.2 U/L,183.8 U/L].The output quantity simulated by MCM was normal distributed.Conclusions When the distribution of the output quantity is normal,the measurement uncertainty evaluated by both MCM and GUM method is nearly the same.When the distribution of the output quantity is unknown,MCM can be used as a verification of GUM method.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. However, emergence of drug resistance limits its potential use. Plumbagin is a natural quinonoid compound isolated from plant. In this study, induced apoptosis effect of the combined treatment with plumbagin and TRAIL on human melanoma A375 cell line was examined and possible mechanism was investigated. The cells were divided into four groups: control group, plumbagin group (plumbagin, 5 or 10 mumol/L), TRAIL group (TRAIL, 30 ng/mL) and plumbagin+TRAIL group (combined treatment group). The apoptosis, and the expression of DR4 and DR5 were detected by flow cytometry. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that the apoptosis rate was 8.3% in TRAIL group, 10.35%-16.94% in plumbagin group and 52.39%-65.39% in combined treatment group, respectively, with the difference being significant between combined treatment group and plumbagin or TRAIL group (P<0.05 for each). Moreover, plumbagin alone could markedly up-regulate DR5 mRNA and protein expression, and slightly increase DR4 mRNA and protein expression. Treatment of human melanoma A375 cells with plumbagin resulted in the activation of Caspase-3, but not Caspase-8. These results suggest that plumbagin might be useful for TRAIL-based treatment for melanoma.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. Epigallocatechin-3-gallate (EGCG) is a polyphenolic constituent of green tea. In this study, inhibitory effect of combined use of EGCG and TRAIL on human melanoma A375 cells was examined and the possible mechanism investigated. The cells were divided into 4 groups: control group, EGCG group (EGCG: 10, 20 mug/mL), TRAIL group (TRAIL: 25 ng/mL) and EGCG+TRAIL group (combined group). The growth inhibition was measured in the A375 cells treated with different concentrations of TRAIL ((25, 50, 75, 100, 125, 150 ng/mL) by MTT assay. The apoptosis was assessed by flow cytometry. The expressions of DR4 and DR5 were detected by flow cytometry and western blotting. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that TRAIL could dose-dependently inhibit the growth of A375 cells and the IC(50) of TRAIL was 150 ng/mL. The apoptosis rate was 11.8% in the TRAIL group, 5%-7% in the EGCG group and 48.9%-59.1% in the combined group. Significant difference was found in the apoptosis rate between the combined group and the EGCG or TRAIL group (P<0.05 for each). The expression of DR4 instead of DR5 was significantly increased in the EGCG group. The activity of caspase-3 rather than caspase-8 was substantially enhanced in the EGCG group. These results suggest that EGCG is useful for the TRAIL-based treatment for melanoma.

Objective: To try to find out the morbidity of elderly chronic pulmonary disease's malnutrition by means of MNA and analyze the consistency between traditionalindex and MNA score. Methods: According to the score of MNA,we divided the 143 elderly pulmonary disease patients into three groups: A group(malnutrition); B group(malnutrition risk);C group(well nourished).The records included anthropometry,biochemical markers,dietary intake as well as the occurrence rate of pneumonia and respiratory failure. Results: There were marked differences on anthropometry,biochemical markers,dietary intake and the occurrence rate of pneumonia and respiratory failure among three groups. And the dietary intake of all three groups did not reach the standard of normal people. Conclusion: MNA is a good method that can be used to evaluate the nutritional state in elderly pulmonary disease patients.

This paper is to introduce our experiences in treating 2 batches of 13 burn victims transferred from remote areas on postburn days 3 and 4. Methods Thirteen burn victims of 2 mass casualties were transferred to our burns institute from remote areas on postburn days 3 and 4 on June 27, 2001 and June 2, 2002, respectively. There were 4 males and 9 females, age ranged from 20 to 43 years, with a mean age of 31.1±6.2 years. The mean total burn area was 74.3%±24.7% TBSA (range, 25% to 97%). Among them, 10 patients suffered from serious burn with mean total burn area involving 86.0%±11.5% TBSA (range, 60% to 97%), and mean full-thickness burn of 63.9%±26.3% TBSA. Four patients also manifested signs of severe inhalation injury, and 6 patients with moderate inhalation injury. In three patients with mean total burn area covering 35.5%±10.0% TBSA (range, 25% to 45%), with mean full-thickness burn of 15.3%±5.0%, were al having moderate inhalation injuries. Among these 13 patients, 3 were having high body temperature (39℃), while 3 manifested hypothermia. The heart rate was 140-160/min, and respiratory rate 26 to 32/min in 6 patients. Abdominal distension or loss of bowel sound were found in 4 patients. Low white cell and platelet count were found in some patients. In 13 cases, liver function, renal function, myocardiac enzyme, and coagulation function were abnormal. Results Among 13 burn victims, one patient died of myocarditis on postburn day 29, and another one died of hepatic failure (history of chronic hepatitis B) on postburn day 45 with only 2% TBSA of burn wound remained open. Conclusion Burns victims occurred in mass casualties who were transferred from remote areas to our Burns Institute were all in critical condition, usually with multiple complications, demanding most meticulous care. Our strategies in this regard consisted of dispatch of experienced surgeons and nurses to the referring hospitals and the airport to receive the patients to offer appropriate care to them during the journey,organization of the medical staff so that each of them was ordained specific function, thus conditions of the patients were evaluated immediately and appropriate treatment started expeditiously for those lethal complications on arrival. Timely and exact comprehensive treatments were prerequisite to save the patients’ life. Adequate metabolic support should be emphasized, and coagulant of anticoagulant treatment should be carried out when indicated.

OBJECTIVETo explore the effects of recombinant human growth hormone (rhGH) when applied postoperatively on the metabolism of branch chain amino acid in severely burned patients.

METHODSFifty burn patients, aged 12 - 50 years and inflicted by more than 30% TBSA with 10% or more of III degree burn and admitted from the January of 1999 to the January of 2001 were enrolled in the study. The patients were randomly divided into rhGH treating (rhGH group) and control groups. Escharectomy was performed within 3 postburn day (PBDs). rhGH (0.3 IU/kg) was injected percutaneously every evening for ten days since the 1st postoperative day (POD). The changes of the plasma levels of GH and branch chain amino acid and the urine level of 3-methyl histidine (3-MH) were observed in the morning in the patients from the two groups.

RESULTSThe plasma GH level before operation decreased obviously in two groups of patients when comparing with normal value (P < 0.05). The plasma GH level in rhGH group was evidently higher than that in control group since the 3rd POD (P < 0.05). There was significant increase of the output amount of urine 3-MH in all patients, but which was obviously higher in control group than that in rhGH group (P < 0.05). The plasma levels of branch chain amino acid in burn patients before and one day after operation were lower than normal levels. The plasma levels of valine, isoleucine and leucine increased to peak values at POD 3 in rhGH group and at POD 7 in control group and decreased thereafter. The plasma branch chain amino acid level in rhGH group was evidently lower than that in control group since POD 7 (P < 0.05). The plasma GH level in rhGH group was negatively and significantly correlated with the urine output amount of 3-MH (P < 0.01).

CONCLUSIONPostoperative application of rhGH in major burn patients might be beneficial to the protein synthesis from amino acids by skeletal muscles and to the decrease of muscle protein degrading rate.

Adolescent ; Adult ; Amino Acids ; metabolism ; urine ; Burns ; blood ; metabolism ; urine ; Child ; Female ; Human Growth Hormone ; blood ; pharmacology ; Humans ; Male ; Middle Aged ; Muscle, Skeletal ; drug effects ; metabolism ; Recombinant Proteins ; blood ; pharmacology

Objective To investigate the impacts of treating stratege for non-infarct related artery on clinical prognosis in elderly patients with acute myocardial infarction (AMI) after urgent percutaneous coronary intervention (PCI). Methods From Augst 2007 to Augst 2010,a total of 75 elderly AMI patients (aged 75 years and over) were treated by urgent PCI and confirmed as multivessel coronary disease in our hospital. Among them,30 patients received medicine combined with PCI once again (PCI group) and 45 patients received only medicine treatemt (medicine group).The major adverse cardiovascular events (MACE) and results ot coronary angiography after treatment for average one year were compared between the two groups. Results There were no significant differences in the rates of in stent restenosis[1 case(2.2 ％)vs.0 case],late loss of in-segment lumen [5 cases(11.1％)vs.3 cases(10.0％)],stent thrombosis[1 case(2.2％)vs.1 case(3.3％)] and larget vessel revascularization [2 cases (4.4 ％ ) vs.1 case ( 3.3 ％) ] between medicine group and PCI group (x2=0.00,0.00.2.03 and 0.00,all P＞0.05).The propertions of angina recurrence and second hospital admission for heart diseases were lower in PCI group than in medicine group [36 cases (80.0％)vs.14 cases(46.7％),18 cases(40.0％)vs.5 cases(16.7％),x2=9.00,4.61,P＜0.01and P＜0.05].However,no differences were found in the secondary heart failure,recurrent nonfatal myocardial infarction,severe arrhythmia,all- cause death and mortality rate of cardiovascular disease between the two groups (x2 =0.09,0.00,0.00,0.00 and 0.00,all P＞ 0.05). Conclusions Compared with single medicine therapy,the medicine combined with PCI for non- infarct-related artery may decrease the rates of angina recurrence and second hospital admission for heart diseases in elderly patients with AMI.

Background:The occurrence of gastric cancer is a gradual process with multiple factors and multiple steps. In addition to cytological and structural abnormalities,there are abnormal molecular expressions,which involve the activation of many oncogenes and the inactivation of tumor suppressor genes. Aims:To explore the expressions and significance of Ki-67,p53 and P504s in normal gastric mucosa,atrophic gastritis with intestinal metaplasia,low-grade intraepithelial neoplasia,high-grade intraepithelial neoplasia and early gastric cancer. Methods:A total of 44 cases of normal gastric mucosa,44 cases of atrophic gastritis with intestinal metaplasia,41 cases of low-grade intraepithelial neoplasia,38 cases of high-grade intraepithelial neoplasia and 35 cases of early gastric cancer from Jan. 2015 to Dec. 2016 at the Affiliated Drum Tower Hospital of Nanjing University Medical School were collected. Expressions of Ki-67,p53 and P504s were detected by immunohistochemical staining. Results:Compared with normal gastric mucosal tissue,the positivity rates of expression of Ki-67,p53 and P504s in atrophic gastritis with intestinal metaplasia,low-grade intraepithelial neoplasia,high-grade intraepithelial neoplasia and early gastric cancer were obviously increased (P＜0.05). With the increasing of severity of the lesion,the positivity rate gradually increased. Conclusions:The expressions of Ki-67,p53 and P504s are closely related to the occurrence and development of gastric cancer,and are involved in the early process of gastric cancer. The detections of these molecular markers are helpful for determining the severity and trend of the lesion,and beneficial for improving the detection rates of gastric precancerous lesion and early gastric cancer.