The present study was designed to test whether Rho-kinase (ROCK) specific inhibitor fasudil (HA-1077) could contribute to migration and vasculogenesis of endothelial cells differentiated from rat bone marrow mesenchymal stem cells (rBMSCs) in vitro. rBMSCs were separated by gradient centrifugation on lymphocytes separation medium from bone marrow of Sprague-Dawley rats, and were cultured, purified and expanded in vitro. Cells of passage 2 to 3 were induced to differentiate into endothelial lineage cells by HG-DMEM plus EGM-2. These cells were identified as endothelial cells with positive factor VIII and Ulex europaeus agglutinin-1 expressions and DiI-Ac-LDL uptake. HA-1077 and VEGF synergistically promoted cell migration, especially in response to transwell chamber assay. When the cells were cultured on ECMatrix™, they showed cellular protrusions and/or cords of aligned cells resembling primitive capillary-like structures at 8 to 12 h of incubation. HA-1077 promoted cell migration and formation of capillary-like tubes. The length of the total capillary tubes was longer than that in the control group (P<0.05). When the cells were exposed to a combination of VEGF and HA-1077, the number of the capillary-like networks and the stability of tube increased. The results obtained suggest that HA-1077 can promote migration and vasculogenesis of endothelial cells differentiated from rBMSCs in vitro.
1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine ; analogs & derivatives ; pharmacology ; Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Movement ; drug effects ; Endothelial Cells ; cytology ; Mesenchymal Stromal Cells ; cytology ; Rats ; Rats, Sprague-Dawley ; rho-Associated Kinases ; antagonists & inhibitors

Background Oxidative stress plays an important role in the pathogenesis of diabetic complications.Peroxiredoxin 6(Prx6) is a doubly-functional protein.and its ability to eliminate phospholipid hydroperoxides is essential. Objective The aim of this study was to investigate the dynamic expression of Prx6 in the retina of streptozotocin(STZ)-induced diabetes and explore its correlation with the progression of diabetic retinopathy. Methods Diabetes was induced by an intraperitoneal injection of streptozotocin(STZ)in 48 clean Wistar rats.The rats were sacrificed at 1,2,and 4 months after the injection of STZ,and expressions of Prx6 protein and Prx6 mRNA in the retina was determined by immunohistochemistry and reverse transcription polymerase chain reaction.Another 12 matched normal Wistar rats were used as the control group. Results The resuh of immunohistochemistry showed that Prx6 protein was expressed in the cytoplasm of the outer nuclear layer(ONL)and inner nuclear layer(INL)in normal rats,and low expression of Prx6 protein was observed in the ganglion cell layer (GCL).In the first month,Prx6 protein was strongly expressed in the INL and the ONL of diabetic rats.However.two and three months after STZ administration,the expression of Prx6 protein was absent in the retina,showing a considerable difference among different course groups(F=22 967.63,P＜0.05).Furthermore,the expression trend of Prx6 mRNA in the retina was similar to that of the Prx6 protein with a significant difference among different course groups(F=942.84,P＜0.05). Conclusion It is conceivable that normal maintenance of Prx6 expression may be important to the prevention of diabetic retinopathy.We hypothesize that oxidative impairments in the retina that develop over time may partly contribute towards the development of retinal dysfunction,which eventually leads to retinal degeneration during the progressive phase of STZ-induced diabetes in adult rats.

Background Platelet-derived growth facto(PDGF) affectthe proliferation of human lenepithelial cell(LECs),and human LECexpresPDGF-α recepto(PDGFR-α) throughoutheilifetime.The binding of activated PDGF-α receptowith PDGF promotethe synthesiof DNA.Othestudiedemonstrated thasilencing of PDGFR-α by antisense oligodeoxynucleotide(ASODN) inhibitthe growth of RPE cellin proliferative vitreoretinopathy (PVR),buwhethethitechnique ifeasible foLECiunclear.Objective Thistudy wato investigate the effecof the knockdown of the PDGFR-α on the proliferation of human LECin vitro,and to offean experimental basifothe gene therapy of posteriocapsule opacification.MethodHuman LECstrain SRA01/ 04 wacultured in α-MEM containing fetal bovine serum.The cellwere incubated in 6-well platea5 × 104 cells/ well and transfection of ASODN-containing liposome waperformed.The cellwere divided into the blank control group (with blank liposome),PDGFR-α missense oligodeoxynucleotide(MSODN) group (with PDGFR-α MSODN + liposome),0.5 μmol/L PDGFR-α ASODN group (with 0.5 μmol/L PDGFR-α ASODN+liposome) and 1.0 μmol/L PDGFR-α ASODN group (with 1.0 μ mol/L PDGFR-α ASODN+liposome).The morphology of LECwaexamined undean inverse microscope 24 houraftetransfection.The expression of PDGFR-α mRNin the cellwadetected by reverse transcription-PC(RT-PCR).The rate of proliferation (A490) of the cellwaassayed using Mtand the inhibitory rate of PDGFR-α ASODN on proliferation wameasured.The percentage of LECin G1 phase waanalyzed by flow cytometer.ResultThe LECgrew well and exhibited polygonal shape in the blank control group and PDGFR-α MSODN group 24 houraftetransfection.Buin the 0.5 μmol/L and 1.0 μmol/L PDGFR-α ASODN groups,the cellappeared round in shape and the numberof cellwere obviously decreased.The expression of PDGFR-α mRNdetected by RT-Pcdemonstrated highelevel in the blank control group and PDGFR-α MSODN group;however,the PDGFR-α mRNexpression waobviously lowein the 0.5 μmol/L and 1.0 μmol/L PDGFR-α ASODN groups.The A490 value wa0.661 ± 0.036,0.655 ± 0.016,0.529 ± 0.030 and 0.441 ± 0.039 in the blank control group,PDGFR-α MSODN group,0.5 μmol/L PDGFR-α ASODN group and 1.0 μmol/L PDGFR-α ASODN group,respectively,showing significandecline in the 0.5 μmol/L PDGFR-α ASODN group and 1.0 μ mol/L PDGFR-α ASODN group in comparison with the blank control group (F=34.08,P＜0.01).The percentageof LECin G1 phase were (47.73±1.18)％,(49.48±1.09)％,(53.31±1.30)％ and (59.98±0.95) ％ in the blank control group,PDGFR-α MSODN group,0.5 μmol/L PDGFR-α ASODN group and 1.0 μmol/L PDGFR-α ASODN group,showing significandifference among them (F =68.41,P＜0.01),and thain the 0.5 μmol/L PDGFR-α ASODN group o1.0 μmol/L PDGFR-α ASODN group showed significantly increase in comparison with the blank control group (P＜0.05).ConclusionPDGFR-α silencing could inhibithe proliferation of human LECin vitro.

Objective To investigate the litholytic effect of stirring by plastic stem inside biliary tract in large common bile duct (CBD) stone. Methods Forty-five patients with large CBD stones were included in this study (8 with the distal bile duct stricture and 5 with too-small Vater's papilla).Plastic stent of 8.5Fr was inserted into the CBD under the guidance of guide wire in all the 45 patients. Results In 10 of the 45 patients, ERCP showed that there was no stone in CBD 3 months later after stent placement. In 22 cases, repeated ERCP revealed that the stone had become smaller to the extent of more than a half or became stone fragments that were easily extracted by baskets or balloon dilation. Thirteen patients continued to have large stone at the second time ERCP treatment, and a new stent was replaced and follow-up performed by B mode ultrasonography of abdominal region per month. Further endoscopic treatment was performed immediately when the stone became small enough. The percentage of 95.6% (43/45) of large CBD stone were eventually cleared after endoscopic treatment for an average of 2.4 times. There were no severe complications related to ERCP or stent placement. Conclusion Placement of plastic biliary stent is a convenient and effective measure for the treatment of large CBD stones. It is especially appropriate for aged patients of CBD stone of high risk.

Objective To explore the relationship between serum C level of C-reactive protein and the classification of acute pancreatitis, and to provide reference for clinical diagnosis and treatment.Methods From January 2012 to January 2015, 100 cases of acute pancreatitis in our hospital were divided into light acute pancreatitis (MAP) group and severe acute pancreatitis (SAP) group.The levels of serum CRP were detected by 1, 3, 5, 7, and levels were compared with the patients of two groups.The sensitivity and specificity of ROC were compared with CRP curve.Results The serum levels of CRP in patients with MAP group and SAP group were increased and then decreased, and serum CRP in MAP group was lower than that in SAP group.The levels of serum CRP in pancreatic necrosis group were lower than those in pancreatic necrosis group.The sensitivity and specificity of CRP were improved with the increase of CRP levels.Conclusion Serum CRP levels can be used for early typing of acute pancreatitis, and it has a certain clinical reference value.

Objective To develop a multiplex PCR-based reverse line blot(mPCR/RLB) hybridization assay to detect,simultaneously,seven genes encoding AR-erm(A/TR),erm(B),mef(A/ E),tet(M),tet(O),aphA-3,aad-6 and two AR-related genes,int-Tn and mreA in group B streptococcus.Methods Nine pairs of specific primers and Oligonucleotide probes targeting erm(A/TR), erm(B),mef(A/E),tet(M),tet(O),aphA-3,aad-6 int-Tn and mreA respectively were modified according to former studies or designed in this study.The primers and probes were labeled with biotin and amino,respectively.The nine genes were amplified simultaneously in the same tube.PCR product hybridized with the probes labeled in the BiodyneC nylon membrane to detect the nine genes.To detect the sensitivity and specificity of the method developed,PCR with single pair of primer targeting each gene were tested in 318 isolates tested and the results were compared with the one abtained by RLB.Results The nine resistance-related genes could be successfully detected by mPCR/RLB assay developed in this study.Based on sequencing,21 of 22 isolates with mef had mef(E)and eight of 353 with int-Tn had an atypical sequence.Except for the above 29 genes,all the others corresponded well with the results obtained by single pair primer PCR.Conclusion The mPCR/RLB assay developed in this study is simple,rapid and suitable for surveillance of antibiotic resistance in GBS.

Puff-balls is a very distinctive macro-fungi in biodiversity.It is used to be a styptic,repellent and alexipharmic in China.This paper simply summarizes the puffball′s taxonomy in the fungi and its distribution in the world; and emphasizes on its composition,food uses,medical uses,herbalism studies and progress of researches on clinical applications.And it points out the patent prospect and current problems of the puff-balls.

Objective To evaluate the application of fibrobronchoscopy in the management of severe brain injury with concurrent pulmonary infection. Methods Forty-three procedures of fibrobronchoscopic sputum clearance or bronchoalvelar lavage were performed in 26 patients with severe brain injury and concurrent pulmonary infection. The effect of fibrobronchoscopic treatment was assessed by observing the changes in the respiratory symptoms, arterial oxygen saturation (SaO2) and partial pressure of oxygen (PaO2) in these patients. Results The symptoms of the respiratory system, SaO2, and PaO2 of the 26 patients were significantly improved after fibrobronchoscopic treatment, which caused no serious complications. Conclusion Fibrobronchoscopy can be safely and effectively used in patients with severe brain injury complicated by pulmonary infection.

OBJECTIVETo identify the major components of traditional Chinese medicine Naodesheng tablet.

METHODSA HPLC-DAD-MS(n) based method was developed to analyze and identify the major components of Naodesheng tablet. Separation was performed on an Agilent Zorbax SB-C(18) column (4.6 mm X 250 mm, i.d, 5 μm) with mobile phase consisting of water with 0.05 % formic acid and acetonitrile as gradient eluent at the flow rate of 0.5 ml.min(-1).

RESULTSA total of 43 components were detected, among which 22 were identified by comparing their UV absorption profiles, the information of molecular Glucosyl puerarin weights, and structures provided by ESI-MS(n) with those of available standards and reference data, such as Safflor yellow A, 4'-O-Glucosyl puerarin, 3'-hydroxypuerarin, Genistein-8-C-apiosyl (1-6) glucoside, Puerarin, 6"-O-xylosyl puerarin, 6"-O-apiosyl puerarin, 3'-methoxy puerarin, 3'-methoxy-6"-o-xylosyl puerarin, Daidzin, Genistin, Pueroside A, Notoginsenoside R(1), Ginsenoside Re, Ginsenoside Rg1,Daidzein,Biochanin A,Ginsenoside Rb(1), Ginsenoside Rc, Ginsenoside Rb(2), Ginsenoside Rb(3), Ginsenoside Rd.

CONCLUSIONThe proposed method can identify the main components of Naodesheng tablet and provide information for the quality control of this medicine.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Ginsenosides ; analysis ; Isoflavones ; analysis ; Mass Spectrometry ; methods

1H NMR metabonomics approach was used to reveal the chemical difference of urine between patients with Xiao-Chaihu Tang syndrome (XCHTS) and healthy participants (HP). The partial least square method was used to establish a model to distinguish the patients with Xiao-Chaihu-Tang syndrome from the healthy controls. Thirty-four endogenous metabolites were identified in the 1H NMR spectrum, and orthogonal partial least squares discriminant analysis showed the urine of patients with Xiao-Chaihu Tang syndrome and healthy participants could be separated clearly. It is indicated that the metabolic profiling of patients with Xiao-Chaihu Tang syndrome was changed obviously. Fifteen metabolites were found by S-pot of OPLS-DA and VIP value. The contents of leucine, formic acid, glycine, hippuric acid and uracil increased in the urine of patients, while threonine, 2-hydroxyisobutyrate, acetamide, 2-oxoglutarate, citric acid, dimethylamine, malonic acid, betaine, trimethylamine oxide, phenylacetyl glycine, and uridine decreased. These metabolites involve the intestinal microbial balance, energy metabolism and amino acid metabolism pathways, which is related with the major symptom of Xiao-Chaihu Tang syndrome. The patients with Xiao-Chaihu Tang syndrome could be identified and predicted correctly using the established partial least squares model. This study could be served as the basis for the accurate diagnostic and reasonable administration of Xiao-Chaihu-Tang syndrome.
Humans ; Least-Squares Analysis ; Medicine, Chinese Traditional ; Metabolome ; Metabolomics ; Proton Magnetic Resonance Spectroscopy ; Syndrome ; Urinalysis