Objective To observe the genetic predisposition of complement C5 gene polymorphisms in proliferative diabetic retinopathy (PDR) in Chongqing Han population.Methods 400 type 2 diabetes (T2D) patients (case group) and 600 age-and sex-matched healthy controls (control group) were enrolled in this study.There were 8 PDR patients in case group.All the subjects were Han ethnic people.The immune-related representative SNP locus of C5 gene including rs2269067,rs7040033,rs7027797 were screened by linkage disequilibrium analysis.Locus rs1017119 was selected by TagSNP and was around the above three loci.Subjects' peripheral venous blood was collected and DNA was extracted.Genotyping was examined by PCR-restriction fragment length polymorphism method.The level of C5 plasma protein was measured by enzyme-linked immunoabsorbent assay.Results The frequency of GG genotype of rs2269067 was significantly increased in PDR patients in cases group compared with controls (Pc=3.4 × 10-5,OR=1.87,95％CI=1.43-2.44;P=3.1 × 10-6).There was no differences in frequency of G,CC and CG genotype of rs2269067 between two groups (P=1.4 × 10-4,1.000,1.0 × 10-6).There were no differences in frequency of G,CC,CG,GG genotype of rs7040033,rs1017119,and rs7027797 between two groups (P＞0.05).The production of C5 plasma protein was significantly increased in case group as compare with control group (P=0.0004).An increased production of C5 plasma protein was observed in rs2269067 GG genotype cases compared to CG or CC cases (P=0.003,0.001).Conclusion C5 rs2269067 GG genotype may be associated with the PDR of T2D in Chongqing Han population.

Objective To explore the possible effects of nutritional supplements on brain function as reflected by Water Maze test performance in mice after +Gz exposure.Method Mice were arranged into control group (group A),+Gz group without nutritional supplementation (group B) and +Gz plus nutritional supplementation group (group C).Each group contains 12 mice.Mice in group A were not exposed to +Gz while mice in both group B and group C were exposed to 8 min +10 Gz.Distilled water was gavaged to group B mice 3 h before +Gz exposure.On the day before +Gz exposure pyridoxol fortified water was given and 3 h before exposure mixed amino acids solution were gavaged to group C mice.Water Maze test was done and scores were recorded in all groups.After the Water Maze test was completed,blood was collected through the eyes for serum amino acid determinations and brain tissue was collected by decollation for monoamine determination and γ-glutamyl transferase (GGT) activity evaluation. Result After +Gz exposure,longer completion time and more mistakes were observed in Water Maze test in group B as compared with group A and a trend of improvement in group C was noticed. The ratio of brain 5-HT to dopamine(DA) was significantly reduced in group C as compared with group B.Gamma glutamyl transferase (GGT) activity in brain tissue in group C and group B increased significantly. Conclusion High sustained +Gz exposure significantly reduces Water Maze test performance in mice (longer completion time and more mistakes).It seems that there is a trend of improvement in Water Maze performance in mice in dietary nutritional supplementation group,which might be due to significant reduction in ratio of brain 5-HT to DA in mice with nutritional supplementation.

Objective To investigate effects of ulinastatin preconditioning on cerebral ischemia-reperfusion injury in patients undergoing operation on aorta with deep hypothermic circulatory arrest.Methods 30 patients aged 30-50 with national institutes of health stroke scale(NIHSS) ＜ 10 undergoing operation on aorta with deep hypothermic circulatory arrest,were randomly divided into 2 groups(n =15):normal saline control group(group C),ulinastatin preconditioning group(group U).In group U,ulinastatin 20 000U/kg was infused via central vein at 500-1000 U · kg-1 · min-1 from after tracheal intubation,until 10 min before ascending aortic cross-clamping.In group C,same volume normal saline was infused instead of ulinastatin.Blood samples were taken from internal carotid vein at 5 min before the beginning of deep hypothermic circulatory arrest(T1),15 min after the beginning of deep hypothermic circulatory arres(T2)and 15 min after the end of deep hypothermic circulatory arrest(T3)for determination of plasma concentrations of S-100β,CK-BB,Glutamate(Glu) 、TNF-α、IL-1 、IL-10、MDA,SOD and TGF-β1.Cerebral funcition was evaluated and scored using NIHSS at 2 day after operation.Results Plasma concentrations of S-100β,CK-BB,Glu,TNF-o、IL-1 and MDA were lower,the levels of SOD,IL-10 and TGF-β1 were higher,and the NIHSS score was lower in group U (P ＜ 0.05).Conclusion Ulinastatin preconditioning can lighten cerebral ischemia-reperfusion injury in patients undergoing operation on aorta with deep hypothermic circulatory arrest.The mechanism is involved in inhibit the formn of reactive oxygen free radical.

Objective To evaluate the effects of ulinastatin on renal ischemia-reperfusion injury in patients undergoing operation on aorta with deep hypothermic circulatory arrest (DHCA ) .Methods Thirty patients ,aged 30-50 yr ,of ASA physical status Ⅲ or Ⅳ (NYHA Ⅱ or Ⅲ) ,scheduled for elective operation on aorta with DHCA ,were randomly divided into 2 groups ( n=15 each) using a random number table :control group (group C ) and ulinastatin group (group U ) .In group U ,ulinastatin 20 000 U/kg was infused via the central vein at 500-1 000 U·kg-1 ·min-1 from the time immediately after tracheal intubation until 10 min before ascending aortic cross-clamping .In group C ,the equal volume of normal saline was infused instead of ulinastatin .At 5 min before the beginning of DHCA (T1 ) and 15 min after the end of DHCA (T2 ) ,blood samples were taken from the extracorporeal circulation for determination of polymorphonuclear leukocyte counts , and plasma levels of intercellular adhesion molecule-1 , tumor necrosis factor-α, iterleukin-6 (IL-6 ) IL-8 , IL-10 , malondialdehyde , myeloperoxidase ,atrial natriuretic peptide ,cystatin C ,and creatinine .Results The polymorphonuclear leukocyte counts and plasma levels of intercellular adhesion molecule-1 , tumor necrosis factor-α, IL-6 , IL-8 , malondialdehyde , myeloperoxidase , cystatin C , and creatinine were significantly lower , and the plasma concentrations of IL-10 and atrial natriuretic peptide were higher in group U than in group C ( P< 0.05 ) . Conclusion Ulinastatin can attenuate renal ischemia-reperfusion injury in patients undergoing operation on aorta with DHCA and inhibition of inflammatory responses is involved in the mechanism .

OBJECTIVE:To study the purification technology of total flavonoids from Nelumbinis receptaculum by macropo-rous resin. METHODS:Using adsorption rate and desorption rate of total flavonoids from Nelumbinis receptaculum as index,the type of macroporous resin was selected by static adsorption-desorption tests;With adsorption rate of total flavonoids as index,sin-gle factor test was used to investigate the effects of the concentration of total flavonoids,adsorption time,adsorption speed, drug-loading amount,water amount,volume fraction and amount of eluant and other factors on the purification technology. The op-timal technology was validated. RESULTS:Among 10 kinds of resin,HPD-400 macroporous resin was found to have the best ad-sorption and desorption effects. The optimal purification conditions was as follows as the concentration of total flavonoids 7.00 mg/ml, adsorption time of 3 h,flow rate for sampling of 3 column volume (BV)/h,drug-loading amount of 8 BV,water amount of 6 BV,50% ethanol elution amount of 4 BV. In validation test,mass fraction of total flavonoids from purified Nelumbinis receptacu-lum were 63.88%,62.50% and 63.44%(RSD=1.11%,n=3). CONCLUSIONS:HPD-400 macroporous resin could purify total flavonoids from purified Nelumbinis receptaculum,and established purification technology is stable and practical.

Objective To evaluate the effects of ulinastatin preconditioning on protamine-induced pulmonary injury in patients undergoing cardiac valve replacement under cardiopulmonary bypass (CPB).Methods Sixty NYHA class Ⅱ or Ⅲ and ASA physical status Ⅱ or Ⅲ patients of sexes,aged 21-59 yr,scheduled for elective cardiac valve replacement under CPB,were randomly divided into 3 groups (n =20 each):group control one protamine given via central vein (group C1) ; group control two protamine given via ascending aorta (group C2) ;group ulinastatin preconditioning (group U).Heparin was neutralized with protamine after termination of CPB.Ulinastatin 20 000 U/kg was infused via the central vein at a rate of 500-1000 U· kg-1 · min-1 starting from the time point after tracheal intubation until 10 min before cross-clamping of superior vena cava and inferior vena cava in group U.At 10 min after termination of CPB,protamine 4 mg/kg was infused over 8 min via the right internal jugular vein in groups C1 and U,or via the aortic root in group C2.Blood samples were obtained from the left atrium and right atrium at 5 min before neutralization of heparin with protamine (T1) and 15 min after neutralization of heparin with protamine (T2) for determination of polymorphonuclear leukocyte (PMN) and platelet (Plt) counts,and plasma concentrations of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1α (6-keto-PGF1α).Blood samples were obtained from the left atrium at T1 and T2 for determination of the levels of TNF-α,IL-1,IL-8,CD11b/CD18,C3a,C5a,and malondialdehyde (MDA) and superoxide dismutase (SOD) activity and for blood gas analysis.Alveolar-arterial oxygen gradiant (A-aDO2),respiratory index (RI) and oxygenation index (OI) were calculated.Pulmonary arterial pressure (PAP) was recorded.Results Plt and PMN counts in the blood obtained from the left atrium were significantly lower,and plasma TXB2 concentrations in the blood obtained from the left atrium were higher at T2 in group C1,and the plasma 6-keto-PGF1α concentrations and SOD activity in the blood obtained from the left atrium were higher at T2 in groups C2 and U than those in the blood obtained from the right atrium (P ＜0.05).Compared with group C1,Plt and PMN counts and plasma 6-keto-PGF1α concentrations were significantly increased,the levels of plasma TXB2,TXB2/6-keto-PGF1α,TNF-α,IL-1,IL-8,C3a,C5a and MDA were decreased,CD11b/CD18 expression was down-regulated,PAP,A-aDO2 and RI were decreased,and OI was increased at T2 in C2 and U groups (P ＜ 0.05).There were no significant differences in the parameters mentioned above between groups C2 and U (P ＞ 0.05).Conclusion Ulinastatin preconditioning can inhibit protamine-induced pulmonary injury in patients undergoing cardiac valve replacement under CPB,and the effect is similar to that of protamine administered via the aorta.

Objective To evaluate the effect of ischemic postconditioning on myocardial injury in the patients undergoing cardiac valve replacement under cardiopulmonary bypass (CPB).Methods Thirty patients of both sexes,aged 21-59 yr,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ (New York Heart Association Ⅱ or Ⅲ),with left ventricular ejection fraction ≥ 40％,scheduled for elective cardiac valve replacement under CPB,were divided into 2 groups (n=15 each) using a random number table:control group (C group) and ischemic postconditioning group (P group).In group P,ischemic postconditioning was induced by 3 cycles of 30 s aortic unclamping followed by 30 s cross-clamping starting from 5 min before complete opening of the ascending aorta.After induction of anesthesia and before operation (T1),at 1 h after opening of the ascending aorta (T2),at the end of operation (T3),and at 12,24 and 48 h after opening of the ascending aorta (T4-6),blood samples were taken from the radial artery for determination of plasma concentrations of heart-type fatty acid-binding protein (H-FABP),ischemia-modified albumin (IMA),and cardiac troponin Ⅰ (cTnI).Myocardial specimens in the right auricle were obtained at T2,and the pathological changes were examined using an optical microscope.Results The concentrations of plasma cTnI at T2-6 and H-FABP and IMA at T2-5 were significantly higher than those at T1 in the two groups (P＜0.05).Compared with group C,the concentrations of plasma cTnI at T2-6,H-FABP at T2-4,and IMA at T2,3 were significantly decreased (P＜0.05),and the pathological changes of myocardial tissues were significantly reduced in group P.Conclusion Ischemic postconditioning can reduce myocardial injury in the patients undergoing cardiac valve replacement under CPB.

Objective To construct and identify retroviral-mediated short hairpin RNA ( shRNA ) expression vectors of ERβ419, and explore ERβ419 unknown biological function in beagles in future.Methods To screen out the most effective gene silencing sequence of beagle ERβ419 mRNA using qRT-PCR and Western Blot assays, imitate beagle estrogen target cells.Results qRT-PCR results showed, ERβ419-shRNA1 ( P <0.01 ) and ERβ419-shRNA3 ( P <0.01)differed significantly, Western Blot result as same as qRT-PCR,ERβ419-shRNA3 is the best choice.Conclusion Beagles ERβ419-shRNA3 retrain most effectively target gene repression. It is applied to explore ERβ419 unknown biological function in beagles reproductive system, and to prevent and treat beagles reproductive function diseases.

ObjectiveTo investigate the effects of ulinastatin postconditioning and combining ulinastatin postconditioning with pretreatment on myocardial inflammatory response in patients undergoing cardiac valve replacement under CPB.MethodsEighty NYHA class Ⅱ or Ⅲ patients of both sexes aged 21-59 yr undergoing cardiac valve replacement under CPB were randomly divided into 4 groups ( n =20 each): group control (group C) ; group ulinastatin pretreatment ( group U1 ) ; group ulinastatin postconditioning (group U2 ) and group ulinastatin pretreatment and postconditioning combined (group U3 ).Ulinastatin 20 000 U/kg was infused via central vein at 500-1000 U·kg-1 ·min-1 after tracheal intubation until 10 min before cross-clamping of ascending aorta in groups U1 and U3.Ulinastatin 10 000 U/kg was infused into root of aorta at 4000-5000 U· kg- 1 · min- 1 at 5-7 min before declamping of aorta in groups U2 and U3.Blood samples were obtained from radial artery before cross clamping of ascending aorta,at 40 min after aortic cross-clamping,at 45 min after declamping of aorta (T3) and at the end of operation for polymorphonuclear leukocyte (PMN) count,routine analysis of blood and determination of plasma concentrations of IL-10,TNF-α,IL-1 and IL-6 (by ELISA).Myocardial specimens were obtained at 45 min after declamping of aorta for determination of IL-1β and IL-6 expression by immune-histochemistry.Results Ulinastatin pretreatment and/or postconditioning significantly increased plasma IL-10 concentration and decreased plasma IL-1,IL-6,TNF-α concentrations and PMN count and myocardial IL-1β and IL-6 expression in groups U1,U2 and U3 as compared with group C.Plasma IL-10 concentration was significantly higher and plasma IL-1,IL-6 and TNF-α concentrations,PMN count and myocardial IL-1β and IL-6 expression were lower in group U3 than in groups U1 and U2.ConclusionUlinastatin postconditioning can inhibit myocardial imflammatory response in patients undergoing valve replacement under CPB.The protective effect can be augmented by combining ulinastatin postconditioning with pretreatment.

This paper reports the diagnosis and therapy of one case of Dipylidium caninum infection in an infant.