Object To establish the process of purification for ?-linolenic acid from the fruit oil of Camptotheca acuminata Decne.. Methods Complexometry by AgNO 3 was applied. Results The optimal conditions: the AgNO 3 concentration was 4 mol/L, the complexometric temperature was lower than 15 ℃, and the complexometric time was 2 h. The purity of ?-linolenic acid was 91.25%. Conclusion The concentration of ?-linolenic acid from the fruit oil of C. acuminata can reach to 45.8%, therefore it is a new abundant resource for ?-linolenic acid. ?-linolenic acid can be well purified in the fruit oil by this process.

Objective Our retrospective study is aimed to discuss the relationship between the curve changes of T lymphocyte subsets before and after the operation and the infectious complications after radical resec -tion of gastric cancer .Methods Clinical data of patients with gastric cancer received treatment at Central Hospi -tal of Panjin City,Liaoning Province from 2012 to 2014 were retrospectively analyzed .The enrolled patients were divided into two groups according to the infectious complications ,observe group ( patients with infectious complica-tions) and control group ( patients without infectious complications ) .The results of T lymphocyte subsets in two groups were compared .Results A total of 70 patients was retrospective analyzed , including 33 pationts in ob-serve group and 37 pationts in control group.At the 4th day after operation,the levels of CD3,CD4 in the observe group were obviously lower than those in control group .There was no statistical difference at the other time points . At the 4th and 9th day after operation,the levels of CD8,CD4/CD8 rate in the observe group were obviously lower than those in control group .There was no statistical difference at the other time points .Conclusion There is re-markable relationship between the curve changes of T lymphocyte subsets before or after the operation and the in -cidence of infectious complications after radical resection of gastric cancer .

Objective To evaluate the efficacy of endoscopic variceal ligation and somatostatin for esophageal varices bleeding.Methods Eighty patients with hepatic cirrhotic esophageal varices bleeding were allocated into two groups,forty in endoscopic variceal ligation(EVL)group and the others in somatostatin(stilamin)group.Results Initial hemostasis rates(including emergency hemostasis and no bleeding for 72 hours)and one-month rebleeding rates in EVL group and somatostatin group were 97.5% vs 80%(P0.05).The hepatic encephalopathy rates were 2.5% vs 15%(P0.05).Conclusion Endoscopic variceal ligation is superior to somatostatin for hepatic cirrhotic esophageal varices bleeding.EVL can lower the rate of hepatic encephalopathy.

Objective To evaluate the therapeutic effect of early fluid resuscitation under the guidance of pulse indicator continuous cardiac output (PiCCO) on patients with severe acute pancreatitis (SAP).Methods Clinical data of 18 SAP patients (research group),who had undergone fluid resuscitation under the guidance of PiCCO in the Department of Critical Care Medicine of the Second Affiliated Hospital of Anhui Medical University from October 2011 to October 2013,were analyzed prospectively.At the same time,clinical data of 25 cases (control group) that had undergone fluid resuscitation without the guidance of PiCCO from January 2009 to September 2011 were collected retrospectively.The volume of fluid and clinical data were compared between two groups.Results During the first 6 hours,0-24 hours,24-48 hours,and 0-72 hours after intensive care unit (ICU) admission,the research group received larger volume of fluid than that of the control group (mL:2 133 ± 1 593 vs.1 024 ± 421,t=3.337,P=0.002; 5 960 ±2 951 vs.3 767 ± 854,t=3.531,P=0.001; 4 709 ± 1 508 vs.3 863 ± 1 122,t=2.112,P=0.031 ; 14 601 ± 5 095 vs.11 409 ± 2 667,t=2.673,P=0.007).Compared with the control group,the incidence of application of blood purification was lowered [5.56％ (1/18) vs.44.00％ (11/25),x2=7.688,P=0.006],the duration of the systemic inflammatory response syndrome (SIRS) was shortened (days:3.54 ± 2.44 vs.5.62 ± 3.62,t=2.113,P=0.041),acute physiology and chronic health Ⅱ (APACHE Ⅱ) score was significantly declined at 24 hours after admission (11±4 vs.14 ± 5,t=2.104,P=0.042),the blood lactic acid was decreased more significantly after 72 hours (mmol/L:3.10 ±0.55 vs.2.40 ± 1.12,t=2.442,P=0.019),and the length of ICU stay was shortened (days:10 ±9 vs.20 ± 10,t=3.371,P=0.002) in research group.But there was no significant difference in the percentage of the use of vasoactive drugs [16.67％ (3/18) vs.24.00％ (6/25),x2 =0.340,P=0.560],the incidence of invasive mechanical ventilation [50.00％ (9/18) vs.52.00％ (13/25),x2 =0.017,P=0.897],72-hour urea nitrogen changes (mmol/L:-0.33 ± 4.71 vs.-0.09 ± 5.37,t=0.152,P=0.880),and the percentage of abdominal infection [16.67％ (3/18) vs.16.00％ (4/25),x2=0.003,P=0.953] between research group and control group.The mortality in research group was lower than that in control group [5.56％ (1/18) vs.20.00％ (5/25)] without statistical difference (x2=1.819,P=0.178).According to the 2012 Atlanta classification,patients were re-evaluated after 48 hours fluid resuscitation.Six patients in research group developed moderately severe acute pancreatitis,and the incidence was significantly higher than that in control group [33.33％ (6/18) vs.8.00％ (2/25),x2=4.435,P=0.034].The time of mean PiCCO installation was 4.5 days in 18 cases of the research group,and no related complications occurred.Conclusions The PiCCO device may be a useful adjunct for fluid resuscitation monitoring in patients with SAP within 72 hours.Early fluid resuscitation under the guidance of PiCCO may be helpful in improving tissue perfusion,reducing the application of blood purification,as well as shortening length of ICU stay.This program did not increase the risk of invasive mechanical ventilation,and no obvious change in mortality rate was observed.

Objective To observe the effect of apolipoprotein E mimetic peptide (ApoE23) on lipopolysaccharide (LPS) levels in plasma and the regulatory role of ApoE23 on low density lipoprotein receptor (LDLR) on liver cells in the septic mice.Methods An ApoE mimetic peptide was designed and referred terminologically as ApoE23 in abbreviation.ApoE23 was synthesized by using solid phase synthesis assay and were refined by using high performance liquid chromatography (HPLC).The peptide was identified and confirmed by using electron spray ionization mass spectrometry and amino acid composition analysis.The C57BL mice infected with Salmonella typhimurium group B were treated with apoE23 injected into tail vein.The plasma LPS levels were measured by using immunoturbidimetry.The LDLR expression and level on liver cells were measured by real time PCR and western blot respectively.Results The plasma LPS levels significantly increased and the liver LDLR expression decreased in the septic mice.ApoE23 treatment markedly reduced the plasma LPS levels and redressed the LDLR down-expressions on liver cells both in mRNA and protein levels compared to the septic mice without ApoE23 treatment.Conclusions The reduction of LPS level after ApoE23 treatment may be associated with the modulation role of ApoE23 in LDLR expression on liver cells,and ApoE23 may be a potential agent against bacterial sepsis as well.One of possible mechanisms was most likely associated with effect of ApoE23 on LDLR expression.

Objective: To clone the Fab gene of a monoclonal antibody (mAb) BDI against human bladder cancer and its expression in E. coli. Methods: Fd and K genes of mAb BDI were cloned by RT-PCR and inserted into an Fab expression vector. Phage displaying Fab and soluble Fab were expressed in E. coli. The N-terminal sequence of VH region was corrected by PCR mediated mutagenesis. The antigen-binding characteristics of the Fab were tested by ELISA and immu-nohistochemistry. Results: Fd and K genes were cloned into the expressing vector p3MH and the phage displaying antibody and soluble Fab were expressed in E. coli, which showed weak binding activity to bladder cancer cells. Correction of the N-terminal sequence of the VHimproved the biding activity dramatically. The feasibility of the application of the Fab in phage antibody library screening was confirmed by a simulated panning procedure. Conclusion: The Fab gene of an anti-human bladder cancer mAb was expressed in E. coli. The importance of the N-terminal sequence on antibody binding activity was suggested.

Objective To explore the differential liver plasma membrane (PM) proteins that may be related to the occurrence,development and reversal process of hepatitis and to understand the pathogenesis of hepatitis and the new drug targets by performing a comparative proteomics research of liver PM between hepatitis B surface antigen (HBsAg) transgenic mice and wild-type C57 mice.Methods A 6-month-old HBsAg transgenic mouse model was established.The pathological examination was performed to observe the pathological changes of transgenic mice and wild-type C57 mice.The PM from liver tissue of 6-month-old transgenic mouse and the control mouse were purified through twice sucrose density grade centrifugation combined with second antibody magnetic bead enrichment.The purity of extracted PM was verified by Western blot.Differential proteome expression analysis was performed by using two-dimensional electrophoresis (2-DE) and ImageMaster software analysis.The differentially expressed proteins were lysed by trypsin and identified by liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) analysis.Results The pathological examination results showed that hepatitis was observed in the transgenic mouse group,while no abnormity was found in the controls.The PM was successfully enriched and the mitochondria contamination was reduced by sucrose density grade centrifugation combined with second antibody magnetic bead purification treatment.Thirty differential mice liver PM protein spots were visualized,in which 11 non-redundant proteins were successfully identified by LC-MS/MS in transgenic mouse group,including 9 up regulated protein spots and 2 down-regulated protein spots.These differentially expressed proteins included keratin,cardiac Ca2+ release channel,cytochrome B5,ATP synthase subunit alpha,etc.Conclusions A batch of HBsAg gene expression related differential proteins are identified in mouse liver plasma.These proteins might be new drug targets for anti-HBV treatment.This study will guide further investigation on the mechanism of HBV infection induced hepatitis.

Objective To investigate the effects of soluble endoglin(sEng)on nitric oxide (NO)production and endothelial nitric oxide synthase(eNOS)phosphorylation in cultured human umbilical vein endothelial cells.Methods Human umbilical vein endothelial cells within 3 passages seeded in culture plates of 96 wells,were stimulated by total culture medium(control group)or sEng (1,10 and 100 μg/L)respectively.Cells and medium were collected after cells were cultured for 6,12 and 24 hours respectively.The concentration of the metabolites of NO in each group was measured by nitrate reductase method.The expression of eNOS and eNOS-Ser(p)1177 were detected by Western blot.The expression of eNOS mRNA in each group was detected by real-time fluorescence reverse transcription-polymerase chain reaction.Analysis of variance,LSD method and pearson correlation were used to compare the difference between groups.Results(1)The concentration of the metabolites of NO in 1,10 and 100μg/L sEng groups was(59.25±1.63),(41.08±2.71)and (30.38±1.63)μmol/L respectively after cultured for 6 hours;(54.98±3.34),(35.00±8.60)and (19.82±3.75)μmol/L for 12 hours; and(46.14±4.93),(30.24±2.08)and(12.78±5.01)μmol/L for 24 hours.There was no significant changes in control group with time going by(F=2.30,P=0.14).The concentration of the metabolites of NO was significantly lower in sEng group,and which had negative correlation with culture time(r=-0.98,P＜0.05)and dose(r=-0.88,P＜0.05).(2)The expression of eNOS in 1,10,100 μg/L sEng groups was 0.71 ± 0.00,0.47 ± 0.00 and 0.32±0.00 after cultured for 6 hours; 0.58±0.00,0.42±0.00 and 0.25±0.00 for 12 hours; and 0.49±0.00,0.33±0.00 and 0.18±0.00 for 24 hours.While the expression of eNOS and eNOS-Ser (p)1177/eNOS had no significant changes in control group with time going by(F=3.59 and 0.37,P=0.09 and 0.80).The expression of eNOS protein and eNOS-Ser(p)1177 decreased significantly in sEng groups,which had negative correlation with culture time(r=0.98 and-0.96,P＜0.05)and dose(r=-0.76 and-0.79,P＜0.05).(3)The expression of eNOS mRNA decreased significantly in sEng groups.Which also had negative correlation with culture time(r=-0.51,P＜0.05)and dose(r=-0.82,P＜0.05).Conclusions sEng might inhibit eNOS activity by blocking 1177 Ser phosphorylation to decrease NO production.

Objective To investigate the short axis systolic function of children with single-left ventricle(LV),and to appraise the clinical value of velocity vector imaging(VVI) on assessing it.Methods The study group consisted of 14 patients with single-LV.The control group consisited of 14 age-matched normal children.VVI was used to analyse the circumferential and radial strain and strain rate of regional single-LV at the level of papillary muscle.Results Compared with values in control group,single-LV circumferential strain values were significantly lower in anterior septum,posterior septum,anterior wall,lateral wall and inferior wall(all P ＜0.05).Single-LV circumferential strain rate values were significantly lower in anterior septum,posterior septum,anterior wall,lateral wall and posterior wall compared with controls(all P ＜0.05). In control group,circumferential strain and strain rate were higher in inferior septum and anterior septum compared with other segments( P ＜0.05).Single-LV radial strain values were significantly lower in all segments compared with values in control (P ＜ 0.05).In control group,radial strain and strain rate of papillary muscle level showed no significantly difference( P ＞0.05).Conclusions Circumferential and radial systolic ventricular function are impaired in children with single-LV.VVI can beused as a quantitative tool in evaluating the short axis systolic function of single-LV.

Objective: To investigate sodium and potassium intakes among individuals undergoing physical examinations in Changsha City, so as provide the evidence for developing nutritional interventions. Methods: The individuals undergoing physical examinations in the Center for Health Management, The Third Xiangya Hospital of Central South University from February 2017 to March 2020 were selected, and their gender, age, history of diseases, history of medications, smoking and drinking behaviors were collected through questionnaire surveys. The blood pressure, height and body weight were measured, and the body mass index (BMI) was calculated. The levels of blood glucose, total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and serum creatinine were detected, and the 24-h urinary excretions of sodium and potassium were measured using the Kawasaki method to estimate sodium and potassium intakes. According to the recommended intakes for preventing chronic diseases (PI values) in the Chinese Dietary Reference Intakes (2013 revised version), the gender-, age- and BMI-specific intakes of sodium and potassium were analyzed. Results: Totally 50 543 subjects were enrolled, with a mean age of (45.64±11.89) years. There were 28 555 men (56.50%) and 21 988 women (43.50%), and there were 19 405 overweight individuals (38.39%), 6 276 obese individuals (12.42%), 13 410 smokers (26.53%), 14 740 drinkers (29.16%), 9 217 individuals with hypertension (18.24%), 3 589 individuals with diabetes (7.10%), 18 232 individuals with dyslipidemia (36.07%). The mean sodium intake was (4 143.29±1 216.46) mg/d among the study subjects, and there were 49 277 participants with sodium intakes that exceeded the PI value (97.50%). Higher sodium intakes were found in men [ (4 345.52±1 231.45) mg/d ], individuals at ages of 36 to 45 years [ (4 185.41±1 202.95) mg/d ], obese individuals [ (4 578.94±1 282.66) mg/d ], smokers [ (4 191.71±1 219.91) mg/d ], drinkers [ (4 325.12±1 245.65) mg/d ], individuals with hypertension [ (4 358.89±1 265.70) mg/d ] and individuals with dyslipidemia [ (4 261.60±1 228.86) mg/d ]. The mean potassium intake was (1 986.26±526.42) mg/d among the study subjects, and there were 50 440 participants with potassium intakes that exceeded the PI value (99.80%). Higher potassium intakes were seen in men [ (2 061.94±556.91) mg/d ], individuals at ages of 35 years and below [ (2 027.24±533.37) mg/d ], obese individuals [ (2 133.27±580.25) mg/d ], smokers [ (2 023.08±557.72) mg/d ], drinkers [ (2 048.43±557.53) mg/d ], individuals without hypertension [ (1 994.65±552.63) mg/d ] and individuals with dyslipidemia [ (2 038.22±546.52) mg/d ]. Conclusions Higher sodium intakes and lower potassium intakes than the PI values are found among individuals undergoing health examinations in Changsha City. Health education is recommended to be reinforced to improve sodium and potassium intakes.