[ABSTRACT]AIM:Toinvestigatetheroleofcysteine-rich61(Cyr61/CNN1)inproliferationandmigrationof bone marrow mesenchymal stem cells ( BMSCs ) .METHODS: The lentiviral vector carrying CCN 1 ( Lenti-GFP-CCN1 ) was constructed and then transfected into the rat BMSCs .The cells were divided into non-transfection group , transfection group ( transfected with Lenti-GFP-CCN1 ) and negative control group ( Lenti-GFP ) .The fluorescence intensity of the transfected BMSCs was observed under inverted fluorescence microscope .The effects of CCN1 on the proliferation and mi-gration of BMSCs were detected by MTT assay and scratch wound healing assay .RESULTS:The proliferation of BMSCs transfected with Lenti-GFP CCN1 had no significant difference compared with negative control group and control group .The width/thickness ratio of migrated BMSCs in wound healing was significantly higher in Lenti-GFP-CCN1 group than that in negative control group and control group (P<0.05).CONCLUSION:Exogenous CCN1 promotes the migration of BMSCs.

Objective To explore the clinical features and endoscopic manifestations of eosinophilic gastroente-ritis(EG)in children.Methods A retrospective analysis was conducted of the clinical manifestation,laboratory examination,endoscopy(upper and/or colonoscopy)performance,diagnosis and treatment of 49 cases of patients who were diagnosed as EG in Children's Hospital of Nanjing Medical University from July 2013 to July 2015.Results The common clinical manifestations of EG in children were hematochezia(23 cases),diarrhea(20 cases),vomiting(18 cases)and abdominal pain(15 cases).The ages of children admitted to hospital for the first time ranged from 1 month and 7 days old to 13 years and 7 months old,and the mean age was 59.4 months old,in which most patients were younger than 1 year old,accounting for 38.77％(19 cases)of all.Peripheral blood eosinophilia was present in 20 cases(40.82％)of the patients,and only 6/18 cases(33.33％)of the patients had elevated serum IgE.Upon endoscopic analysis,the lesions involved esophagus(4 cases),stomach(14 cases),duodenum(20 cases),small intestine(15 cases),colon(28 cases)and rectum(6 cases),and the most common manifestation under gastroscopy was mucosal hyperemia edema(27 cases)and erosion(9 cases),while the most common manifestation under colonoscopy was mucosal hyperemia edema(25 cases)and nodular hyperplasia(24 cases).All patients improved with food restriction,in which 8 cases were treated with glucocorticoid while 9 cases with oral Singulair and 9 cases with oral Loratadine.All children with symptoms were alleviated somewhat,but 5 cases of them relapsed after drug withdrawal.Conclusions The clinical manifestations of EG in children varied and were mainly hematochezia,vomiting,diarrhea and abdominal pain.Some patients had the elevated peripheral blood eosinophilia and serum IgE.The most common manifestations under gastroscopy were mucosal hyperemia edema and erosion while the most common manifestations under colonoscopy were mucosal hyperemia edema and nodular hyperplasia.

OBJECTIVE: To select specific DNA aptamer for determining ketamine by FluMag-SELEX. METHODS: Based on magnetic beads with tosyl surface modification as solid carrier and ketamine as target, a random ssDNA library with total length of 78 bp in vitro was compounded. After 13 rounds screening, DNA cloning and sequencing were done. Primary and secondary, structures were analyzed. The affinity, specificity and Kd values of selected aptamer were measured by monitoring the fluorescence intensity. RESULTS: Two ssDNA aptamers (Apt#4 and Apt#8) were successfully selected with high and specific abilities to bind ketamine as target with Kd value of 0.59 and 0.66 μmol/L. The prediction of secondary structure was main stem-loop and G-tetramer. The stem was the basis of stability of aptamer's structure. And loop and G-tetramer was the key of specific binding of ketamine. CONCLUSION FluMag-SELEX can greatly improve the selection efficiency of the aptamer, obtain the ketamine-binding DNA aptamer, and develop a new method for rapid detection of ketamine.
Aptamers, Nucleotide/metabolism* ; DNA ; DNA, Single-Stranded/genetics* ; In Vitro Techniques ; Ketamine/metabolism* ; Oligonucleotides ; SELEX Aptamer Technique/methods*

Objective To evaluate the accuracy and reliability of Diana automated blood grouping analyzer in blood grouping and cross matching.Methods 2 300 patients′ABO and RhD blood groups were examined by conventional tube test and the fully auto-mated blood grouping analyzer and 900 patients′samples were tested using Diana automated blood grouping for blood cross matc-hing,and it was compared with polymatching method.Results The analyzer′s accuracy rate of blood grouping by two methods were 99.87% and 100.00%.The incompatibility occurred in 30 specimens in automatic blood type instrument,in 3 specimens in manual polymatching method.Conclusion The results of Diana automated blood grouping analyzer used for blood grouping and cross matc-hing blood testing are reliable.Its experimental operation is normalized and standardized with an advantage of low incidence of human er-ror.Moreover,the experimental results can be permanently preserved,which provides a convenience to search for medical proof.

OBJECTIVE: To study the effects of Yiqi Huayu Recipe and its decomposed formulas-medicated sera on expressions of bcl-2, Bax and caspase-8 of apoptotic anulus fibrosus cells in rats. METHODS: Immunohistochemical and integral optical density analytic methods were used to observe the effects of Yiqi Huayu Recipe-, Yiqi Recipe-, Huayu Recipe-medicated sera and insulin-like growth factor 1 (IGF-1) on the expressions of bcl-2, Bax and caspase-8 of apoptotic anulus fibrosus cells in rats induced by anti-Fas antibody. RESULTS: As compared with apoptosis group, bcl-2 expression was higher, Bax and caspase-8 expressions were lower in Yiqi Huayu-treated, Yiqi-treated, Huayu-treated and IGF-1 groups (P<0.01). As compared with Yiqi-treated group and Huayu-treated group, Bax expression was lower in Yiqi Huayu-treated group (P<0.05). CONCLUSION: Yiqi Huayu Recipe and its decomposed formulas can delay degeneration of the cervical intervertebral disc, which may be due to its action in regulating the expressions of bcl-2, Bax and caspase-8.

The artificial lung is a technical device for providing life support; it will be put in use when the natural lungs are failing and are not able to maintain sufficient oxygenation of the body's organ systems. From the viewpoint of long-term development, the artificial lung should be permanently implanted in the body, so that it will substitute for the human pulmonary function partially or completely. In this paper, four artificial lung technologies were expounded with reference to the development and research process of artificial lung. They were extracorporeal membrane oxygenation, intravascular artificial lung, implantable artificial lung, and pumpless extracorporeal lung assist. In this paper were described the structure of the four kinds of artificial lung, the working principle, and their advantages, disadvantages and indications. The prospect of artificial lung was evaluated in the light of the data from the existing animal experiments and from the clinical experience of the centers.
Animals ; Artificial Organs ; trends ; Extracorporeal Membrane Oxygenation ; instrumentation ; trends ; Humans ; Lung ; Oxygenators, Membrane ; trends

OBJECTIVE To investigate the effect of Radix Isatidis and its constituents indigo and in?dirubin on two principal subtypes of organic cation transporters(OCT)OCT1,OCT2 in vivo in mice. METHODS Decoction of Radix Isatidis (DRI) 1.6 and 6.4 g · kg-1,granules of Radix Isatidis (GRI) 0.615 and 2.460 g·kg-1,indigo 0.008 and 0.640 mg·kg-1 and indirubin 0.0192 and 1.536 mg·kg-1 were ig given to NIH mice(60 mice per group),twice a day for 5 d. Four control groups were set up,including the vehicle of water,0.5% sodium carboxymethyl cellulose(CMC),additives of sucrose plus dextrin (1.5 g · kg-1)and positive control quinidine(0.025 g · kg-1). Sixty minutes after the last dosing,all the mice were iv given metformin(Met)5 mg·kg-1,and at 1.0,2.5,5.0,7.5,10.0 and 20.0 min after Met iv,10 mice in each group were sacrificed to collect whole blood and kidneys respectively. The right kidney was homogenized for Met accumulation test and the left one used to extract total RNA for analysis of OCT1 and OCT2 mRNA expressions by real-time PCR. The contents of Met in sera and kidneys were quantified by HPLC. Major pharmacokinetic parameters of Met in sera were analyzed by pharmacokinetic software(DAS 2.0). RESULTS There was no significant difference between water control group,0.5%CMC group and sucrose plus dextrin group in any examined item. Compared with vehicle control group (water and 0.5%CMC group),all the related pharmacokinetic parameters in DRI 6.4 g · kg- 1,GRI 2.46 g · kg-1,indigo 0.640 mg · kg-1 and indirubin 1.536 mg · kg-1 groups were changed significantly (P<0.05,P<0.01). The elimination half time (t1/2β) was prolonged 13%-97%,volume of distribution reduced by 13%-72%,clearance(Cl)reduced by 9%-65%,and the area under the concentration-time curve (AUC0-20 min) increased by 13%-135%. AUC0-20 min obtained from renal Met accumulations was significantly increased(P<0.01)while Met uptake by kidney slices was reduced(P<0.05,P<0.01). The expressions of OCT1 and OCT2 mRNA were obviously down-regulated(P<0.05,P<0.01). CONCLUSION The mouse renal OCT1 and OCT2 are significantly inhibited by DRI,GRI,indigo and indirubin. The inhibitory effect of Radix Isatidis on OCT1 and OCT2 probably arises from indigo and indirubin contained.

OBJECTIVE:To study the effects of 10 kinds of nephrotoxic TCM on three main subtypes(Oat1,Oat2 and Oat3) of kidney organic anion transporter(Oats)in mice. METHODS:A total of 1 840 SPF NIH mice were randomly divided into nor-mal control group(isovolumic pure water),probenecid group(30 kg/mg),sodium carboxymethyl cellulose(CMC)group,Pulsa-tillae radix,Corydalis rhizoma,Aconiti kusnezoffii radix,Aconiti radix,Angelicae pubescentis radix,Gleditsiae spina,Polygo-num cuspidatum,Kansui radix,Platycladi cacumen,Aucklandiae radix high and low dose groups. Mice were treated twice a day for 5 d,ig. After 1 h of the last dosing,they were iv given PAH in tail(30 mg/kg). The PAH pharmacokinetic parameters of the kidney homogenate were determined and the PAH intake in kidney tissue at the time point of 1,5,10,15 and 20 min was detect-ed. The PAH in blood was analyzed by DAS 2.0 software. The grouping and dosing were the same as before,after 1 h of the last dosing,kidney slices were made and put into PAH-buffer. The PAH intake of kidney slices was determined. RESULTS:Compared with normal control group,the t1/2β in C. rhizoma high dose group,A. kusnezoffii high and low dose groups,A. pubescentis high dose group,P. cuspidatum high and low dose groups and P. cacumen group were increased;Vd were all decreased in 10 kinds of TCM high and low dose groups;except for A. pubescentis low dose group,G. spian low dose group and K. radix low dose group, the CL was decreased and AUC0-20 min was increased in all other groups,with significant difference (P<0.01 or P<0.05). Com-pared with normal control group,the content of PAH in kidney tissue in P. radix high dose group,C. rhizoma high dose group,A. kusnezoffii high dose group,A. radix high and low dose groups,A. pubescentis high and low dose groups,G. spina high and low dose groups,P. cuspidatum high and low dose groups,K. radix high and low dose groups,P. cacumen high and low dose groups and A. radix high and low dose groups were increased,with significant difference (P<0.01 or P<0.05). Compared with normal control group,the intake of PAH in kidney slices in C. rhizoma high dose group,A. kusnezoffii high and low dose groups,G. spi-na high and low dose groups,K. radix high dose group,P. ca-cumen high and low dose groups and A. radix high dose group were decreased,with significant difference (P<0.01 or P<0.05). CONCLUSIONS:The 10 kinds of nephrotoxic TCM probably induced kidney injury through inhibiting the Oat1,Oat2 and Oat3 of Oats.

OBJECTIVE:To study the protective effects of soluble ornithine cyclase activator cinaciguat(called CIN for short) on human umbilical vein endothelial cells (HUVECs) under the condition of high glucose. METHODS:HUVECs were treated with glucose 33.3 mmol/L to induce oxidative stress injury model. HUVECs were divided into normal group,high glucose group and high glucose+CIN 0.01,0.1,1 and 10 μmol/L groups. The cell viability was detected by MTT assay,and the survival rate of cells was callulated. Besides,HUVECs were divided into normal group,normal+CIN 1 μmol/L group,high glucose group,high glucose+CIN 1 μmol/L group. MDA content and SOD activity were determined by TBA method and xanthine oxidase method;mRNA expression of ICAM-1 and VCAM-1 were determined by RT-PCR. RESULTS:Compared with normal group,survival rate decreased in high glucose group(P<0.05);compared with high glucose group,that increased in high glucose+CIN 0.01,0.1,1 and 10 μmol/L groups,in concentration-dependent manner;there was no statistical significance in survival rate between high glu-cose+CIN 1 μmol/L group and high glucose+CIN 10 μmol/L group (P>0.05). Compared with normal group,MDA content and mRNA expression of ICAM-1 and VCAM-1 increased in high glucose group,while SOD activity decreased(P<0.05);above indi-cators had no significant change in normal+CIN 1 μmol/L group(P>0.05). Compared with high glucose group,MDA content and mRNA expression of ICAM-1 and VCAM-1 decreased in high glucose+CIN 1 μmol/L group,while SOD activity increased(P<0.05). CONCLUSIONS:CIN can protect high glucose-induced HUVECs injury in vitro. The mechanism may be associated with in-hibition of oxidative stress and inflammatory response.

OBJECTIVE: To study the effects of Yiqi Huayu Recipe on the gene expression pattern in normal and apoptotic chondrocytes in the cervical intervertebral disc of rats. METHODS: The intervertebral disc endplates of rats were digested enzymatically. The apoptosis of the chondrocytes was induced by anti-Fas antibody. BiostarR-40s microarray chips were used to investigate the gene expression pattern in chondrocytes in the normal control, apoptosis and Yiqi Huayu Recipe groups. The results were scanned with Scan Array 4000 and analyzed with GenePix Pro 3.0, and then subjected to standardization and ratio analysis. RESULTS: In the apoptosis group, 30 kinds of genes expressed in cervical intervertebral disc chondrocytes were screened out, in which 10 were up-regulated (ratio>2) and 20 down-regulated (ratio<0.5) as compared with the normal control group. On the other hand, in the Yiqi Huayu Recipe group, 97 kinds of genes expressed in cervical intervertebral disc chondrocytes were screened out, in which 44 were up-regulated (ratio>2) and 53 down-regulated (ratio<0.5) as compared with the apoptosis group. CONCLUSION: There are some signal transduction pathways that control the apoptosis of the intervertebral disc chondrocytes. Yiqi Huayu Recipe regulates the gene express of the apoptotic chondrocytes in intervertebral disc. The study gives a further understanding to the mechanism of the cervical spondylosis and enriches the theories of qi and blood of traditional Chinese medicine.