ObjectiveTo establish an accurate method for simultaneous determination of plasma Kyn and Trp by HPLC-UV detection.Methods Kyn and Trp were separated on Agilent Hypersil ODS column using 3-nitrotyrosine as internal standard.The mobile phase consisted of 15 mmol/L sodium acetateacetic acid (pH 5.5):acetonitrile 94∶ 6(v/v) at a rate of 0.8 ml/min.The chromatographic separation was performed at 25 ℃.The eluate was monitored with programmed wavelength setting at 360 nm from 0 to 4 min for Kyn and at 302 nm from 4 to 5 min for Trp.The method was applied to determination of plasma Kyn and Trp in 8 chronic glomerulonephritis,10 idiopathic thrombocytopenic purpura,15 chronic hepatitis B virus patients and 15 healthy controls from September to December in 2010.The differences were compared using ANOVA and SNK methods.Results The retention time of Kyn and Trp were 2.9 min and 4.4 min,respectively.For Kyn,the assay was linear from 0.44 μmol/L to 18.30 μmol/L.For Trp,the linearity was from 3.67 μmol/L to 470.00 μmol/L.The detection limits were 0.014 μmol/L for Kyn and 0.122 μmol/L for Trp,respectively.The within-day CVs were ＜ 3％ and the between-day CVs were ＜ 4％.The mean recoveries yield were in the range of 92.29 to 104.40.The plasma concentrations of Kyn were ( 1.59 ± 0.28),(2.73 ± 0.56),(2.69 ± 0.44) and ( 1.54 ± 0.48 ) μmol/L,the plasma concentrations of Trp were (59.8 ± 10.0),(46.1 ± 11.7),(58.5 ±8.0) and (41.4±13.1) μmol/L,the Kyn/Trp were (0.027 4±0.007 5),(0.061 6 ±0.016 5),(0.046 7 ±0.009 1) and (0.038 3 ±0.007 5)in controls,chronic glomerulonephritis patients,idiopathic thrombocytopenic purpura patients and chronic hepatitis B virus patients,respectively.There were significance difference of Kyn,Trp and Kyn/Trp amony the four groups (F=23.734,8.463,20.921,all P＜0.01).Conclusion The method is simple,fast,and suitable for applicability to clinical measurement.

Objective To observe the peroxidase body growth activated receptor γ (PPARγ) agonist rosiglitazone on acute pancreatitis in mice with hepatic injury and to investigate the mechanism of hepatic injury .Methods Seventy‐two male Kunming mice were randomly allocated into three groups(24 cases for each group):acute pancreatitis group(AP group) ,rosiglitazone group (AP‐ROS group) ,saline group(NS group) .Mice were killed at 6 ,12 and 24 h after induction of acute pancreatitis .Serum amylase , ALT and AST activities were measured .The expressions of NF‐κB and PPARγmRNA were assessed by RT‐PCR .The expressions of NF‐κB and PPARγ protein were assessed by Western blot .Results Compared with NS group ,serum amylase ,ALT and AST levels at each time point significantly increased in AP group(P< 0 .01);serum amylase ,ALT and AST levels in AP‐ROS group were significantly lower than those in AP group(P<0 .01) .Compared with NS group ,the expressions of liver PPARγ mRNA and protein in AP group were markedly lower at 6 h and 12 h(P<0 .05) ,and the expressions of PPARγmRNA and protein in AP‐ROS group were significantly higher than those in NS group and AP group(P<0 .01) .The expressions of liver NF‐κB mRNA and NF‐κB p65 protein in AP group were significantly higher than those in NS group and AP‐ROS group at all time points(P<0 .01) .Con‐clusion There are clear relationships between NF‐κB and hepatic injury in acute pancreatitis .The expressions of PPARγin injuried hepatic decreased .Rosiglitazone can increase the expressions of PPARγand prevent the expressions of NF‐κB in hepatic during the early phase of acute pancreatitis .

Objective To summarize the methods of diagnosis and results of surgical treatment of pancreatic insulinoma. Methods The clinical data of 137 patients with insulinoma treated in our hospital during the past twenty-six years were reviewed retrospectively.Results There were 77 males and 60 females. All of them were characterized by the Whipple′s triad. The sensitivity of ultrasonography, CT and MRI for localization was 35.1%, 67.9% and 58.1% respectively. One hundred and tweenty-six patients underwent operation. Of them, 102 cases had tumor enucleation, 4 cases had pancreaticoduodenectomy, 16 cases had distal panreatectomy, and the other 4 cases had only laparotomy. Of the 122 patients, who underwent resection, the tumor was benign in 118(96.7%) and malignant in 4(3.3%). The diameter of the tumor was less than 2cm in 86.9% of cases. In 98.4% of cases the tumors were single and in 1.6% of cases were multiple. 13.1% of the tumors located in the head, 46.7% in the body, and 40.2% in the tail.Conclusions Whipple′s triad and the measurement of fasting glucose, IRI, IGR, C-peptide, and proinsulin levels contribute to the diagnosis of insulinoma. However, the preoperative tumor localization is still difficult. Tumor enucleation is the technique of choice when feasible. Patients in whom tumor localization is unsuccessful at operation should be carefully evaluated to be certain of the diagnosis, and in general should not undergo blind resection.

Aconitum, as a kind of common traditional Chinese medicine, contains multiple biological active substances, with a very high medicinal value but high toxicity. Its major toxic ingredients are aconitine, mesaconitine and hypaconitine, which are also efficient ingredients. Therefore, the safety of its clinical application has aroused wide attention. With the constant deepening of drug development studies, people want to learn about its toxic mechanism and the regularity of its emergence and development of its toxicology, so as to make a scientific and rational assessment for its safety. Therefore, toxicokinetics and metabonomics have gradually become important content in the new drug assessment. During the development of drug performance, it is crucial to establish a scientific, objective and standardized Aconitum safety evaluation system and correctly assess and utilize its toxicity. Having summarized studies on metabonomics and toxicokinetics of Aconitum drugs in recent years, authors proposed to strengthen the studies on Aconitum drug safety assessment and establish a scientific and standardized safety evaluation system as soon as possible, in order to make the national treasure more useful.
Aconitum ; chemistry ; toxicity ; Animals ; Drugs, Chinese Herbal ; pharmacokinetics ; toxicity ; Humans ; Metabolomics

The purpose of this study is to find out anti-HIV-1 reverse transcriptase (RT)/protease (PR) activity and inhibition of virus replication in cell cultures of novel coumarin analogs and determine their structure-activity relationship. Coumarin derivatives have been demonstrated to inhibit the activity of HIV-1 RT/PR in cell free system. It also shows inhibition effects to HIV-1 replication in cell culture. Based on the Chinese traditional pharmacological characteristics and protein three dimension computer aided design, analogs of tetracyclic dipyranocoumarin were synthesized from natural leading compounds. We studied the relationship of antiviral effects and chemical structures via HIV-1 PR/RT enzyme models and cell culture model system. Seven compounds were designed and tested. Several compounds showed anti-HIV-1 activity in varying degrees, especially V0201 showed much higher anti-HIV-1 activity with 3.56 and 0.78 micromol x L(-1) of IC50 against HIV-1 PR/RT and 0.036 micromol x L(-1) against HIV-1 replication in PBMC cultures. V0201 with a novel structure may be a new leading compound. These new compounds are valuable for development of new anti-HIV drugs in the future.

Objective:To investigate the effect and mechanisms of Phloroglucinol ( PG) on renal ische-mia and reperfusion injury (IRI).Methods:Forty-eight male Wistar rats were divided into 3 groups (16 rats per group):sham operated, saline-treated I/R (I/R), and PG-treated I/R (PG).I/R model:Af-ter removing the right kidney , renal I/R injury was induced by clamping the left renal artery for 45 min followed by reperfusion.The rats were administered with PG (30 mg/kg, intraperitoneally) or saline 15 min before renal ischemia .The blood and kidneys were harvested 6 and 24 h after reperfusion .Renal function and histologic changes of serum creatinine (SCr) and blood urea nitrogen(BUN)were assessed. Malondialdehyde (MDA),catalase (CAT),superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) were measured.Nuclear factor-kapa B ( NF-κB) and caspase-3 in the kidneys were also measured.Results:SCr and BUN were (103.9 ±10.4) μmol/L and (15.2 ±1.0) mmol/L in I/R group, and (81.8 ±13.4) μmol/L and (11.5 ±1.2) mmol/L in PG group 6 h after reperfusion .SCr and BUN were (154.9 ±12.1) μmol/L and (28.1 ±1.4) mmol/L in I/R group, and (103.8 ±5.9)μmol/L和(16.0 ±1.0) mmol/L in PG group 24 h after reperfusion.PG treatment significantly attenua-ted renal dysfunction and histologic damage caused by I /R injury(P<0.05).The I/R-induced elevation in kidney MDA level decreased , where as reduced kidney SOD ,CAT and GSH-Px were increased .What is more , the apoptotic tubular cells , the levels of active caspase-3 ,and active nuclear factor kappa B dra-matically decreased after PG treatment .Conclusion:PG protects murine kidney I/R injury by suppres-sing oxidative stress , inflammation , and cell apoptosis .

BACKGROUND:Our prior experiments have confirmed that 10 μmol/L oxytocin can induce transformation of bone marrow mesenchymal stem cels into myocardial cels. OBJECTIVE: To investigate the effects of Ginsenoside Rh2 on oxytocin-induced transformation of bone marrow mesenchymal stem cels into myocardial cels. METHODS:Rat bone marrow mesenchymal stem cels were isolated by differential adherence method. These isolated cels were randomly divided into five groups. In the blank control group, cels were routinely cultured. In the oxytocin group, cels were cultured with 10 μmol/L oxytocin for 2 consecutive weeks. In the Ginsenoside Rh2 low-, middle-, and high-dose groups, cels were treated with 0.5, 1, 2 μmol/L Ginsenoside Rh2respectively for 24 hours and then with oxytocin for additional 2 consecutive weeks. RESULTS AND CONCLUSION: Optical microscopy showed that compared to the blank control group, some cels in the oxytocin group exhibited an increased soma and some cels grew in clusters and the cel clusters enlarged with the increase in Ginsenoside Rh2 dose. Immunocytochemical staining and western blot analysis showed that cardiac Troponin T and connexin 43 protein expression in the oxytocin, Ginsenoside Rh2 low-, middle-, and high-dose groups were significantly greater than in the blank control group (P < 0.05), and cardiac Troponin T and connexin 43 protein expression in the Ginsenoside Rh2 groups was increased with the increase in Ginsenoside Rh2 dose and was significantly higher than that in the oxytocin group (P < 0.05). Confocal laser scanning microscopy showed that the relative fluorescence intensity of free calcium in the bone marrow mesenchymal stem cels in the oxytocin group was significantly increased after induction by oxytocin for 2 weeks (P < 0.05), while the relative fluorescence intensity in the Ginsenoside Rh2 groups was significantly higher than that in the Ginsenoside Rh2 groups and was positively correlated with the dose of Ginsenoside Rh2. These findings suggest that Ginsenoside Rh2 can obviously promote oxytocin-induced transformation of bone marrow mesenchymal stem cels into myocardial celsin vitro.

PurposeAlteration of the regional cerebral flow and cerebrovascular reserve (CVR) caused by cerebral artery stenosis or occlusion is an essential risk factor for ischemic stroke. This study aims to assess the CVR in patients with severe middle cerebral artery (MCA) stenosis or occlusion by using MR perfusion-weighted imaging (PWI) with CO2 inhalation stress test. Materials and MethodsPWI were performed before and after CO2 inhalation stress on 28 patients with severe middle cerebral artery stenosis or occlusion and 10 healthy volunteers. The regions of interest (ROI) were put on the affected hemisphere of the MCA blood supply area and the contralateral side in both groups. The relative cerebral blood volume (rCBV), relative mean transit time (rMTT), relative cerebral blood flow (rCBF) and CVR were measured.Results① The rCBV, rCBF and rMTT of the affected side were 149.16±33.01, 18.04±5.24, 8.65±1.81 before CO2 inhalation stress, and 156.23±21.60, 23.77±8.77, 8.72±3.01 after CO2 inhalation stress. The rCBV, rCBF and rMTT were signiifcantly increased (t=1.238, 2.561 and 2.647, P<0.05) after CO2 inhalation stress. The rCBV, rCBF and rMTT of the contralateral side were 176.22±40.12, 22.43±5.74, 8.10±3.71 before CO2 inhalation stress, and 198.54±39.87, 27.64±7.22, 8.03±2.97 after CO2 inhalation. The rCBV, rCBF was significantly increased (t=1.780 and 1.665,P<0.05) while rMTT was signiifcantly decreased (t=2.871,P<0.05) after CO2 inhalation.②The CVR of the affected side, contralateral side and control group were 18.9%, 26.8%, and 39.1%, respectively. There were significantly differences (t=1.604, 2.321 and 1.874,P<0.05) between the affected and contralateral side, the affected side and control group, the contralateral side and control group.ConclusionPWI with CO2 inhalation stress test is valuable in assessing the cerebrovascular reserve in patients with severe middle cerebral artery stenosis or occlusion.

Objective To examine the changes of red blood cell levels in the obese and non-obese patients with coronary heart disease (CHD) and its clinical significance. Methods 230 cases of coronary heart disease were selected and divided into the obese group and the nonobese group. Obesity and non-obesity were defined based on the body mass index (BMI I (Y) 28.0kg/m2), or waist-hip ratio (men> 0.9, women> 0.85). In addition, 130 healthy subjects were recruited as controls. The pathological status of coronary lesions was quantitatively analyzed according to the Coronary Vascular Image Segmentation Evaluation Criteria (American Heart Association 1984) and the Gensini scoring system. Results of the changes of both the hemoglobin levels and the red blood cell count in the obese group, the nonobese group with CHD and the control group were compared. Besides, Multivariant Logistic Regression Analysis was applied to assess the correlation between the red blood cells and the coronary artery disease. Results The red blood cell count and the level of hemoglobin in the obese group with CHD was higher than that in the non-obese group with CHD [(4.35 ± 0.55) and (4.13 ± 0.56) 109/L; (136.71 ± 15.87) and (129.96 ± 16.23) g/L, P < 0.05 in both]; the proportion of acute coronary syndrome in the obese group with CHD was higher in the obese group with CHD than that in the non-obese group with CHD (P<0.05); Multivariant logistic regression analysis also showed that the red blood cell count was positively correlated with obesity with CHD.Conclusion The red blood cell count and the level of hemoglobin in the obese group were higher than those in the non-obese group; the increase of red blood cell count and hemoglobin level is one of the independent risk factors for the obese patients with CHD.

BACKGROUND:Bone morphogenetic protein-2 (BMP-2) is a key to bone formation and repair.However,it has some disadvantages such as easy to lose and degrade and difficult to sustain continuous effect.OBJECTIVE:To study the preparation and properties of silk fibroin/chitosan/nano-hydroxyapatite (SF/CS/nHA) scaffold loading BMP-2.METHODS:After silk degumming,dissolution and purification,2％ SF solution was obtained.BMP-2 was dissolved in 2％ CS solution,and then fully mixed with equal volume of SF solution and proper amount of nHA.At last,the SF/CS/nHA scaffold loading BMP-2 was prepared using freeze-drying method as experimental group.The SF/CS/nHA scaffold was soaked in the BMP-2 solution as control group.The scaffold porosity was measured by Archimedes method,the surface morphology of the scaffold was observed by scanning electron microscope,the compressive strength was measured by universal testing machine.Scaffolds in the two groups were soaked in PBS,and the release of BMP-2 was measured by ELISA method at different time points.RESULTS AND CONCLUSION:(1) The scaffolds in the two groups had irregular porous structure,interconnected pores and uneven pore wall.There was no significant difference between the two groups in mean pore diameter,porosity and maximum compressive strength.(2) On the 1st day,the release rate of BMP-2 was 4.63％ in the experimental group,and the release curve increased slowly.After 28 days,the release curve of BMP-2 was transferred to the plateau stage.But in the control group,the release rate of BMP-2 on the 1stday was 58.84％,and it was a significant initial burst release.The release curve increased rapidly,and was transferred to the platform stage on the 10th day.The release rate of BMP-2 release was significantly different between the two groups at days 1,2,4,10 (P ＜ 0.05).These results show that the SF/CS/nHA scaffold loading BMP-2 could sustainably and slowly release BMP-2.