AIM: To provide experimental evidence for gene therapy of thrombophilia disease, we constructed the eukaryotic expression plasmid with human thrombomodulin (hTM) gene and observed the alteration of hTM expression on the surface of human umbilical vein endothelial cells (HUVECs) with and without the reconstructive plasmid. METHODS: The whole expressive fragment of hTM gene was amplified by PCR from human genome. Both hTM gene and pcDNA3.1(+)/neo empty vector was digested by HindⅢ and EcoRⅠ. Two digested fragments were ligated into pcDNA3.1/hTM with T_4DNA ligase. After identifying, the reconstructive plasmid transfected into HUVECs using lipofectin. The hTM antigen on the HUVECs was detected by immunohistochemistry. RESULTS: The hTM reconstructive plasmid was confirmed by double endonuclease redigesting and sequencing. About 10% HUVECs were transfected by pcDNA3.1/hTM plasmid with lipofectin and the high-level hTM was detected on the transfected cells. CONCLUSION: We constructed the pcDNA3.1/hTM plasmid successfully, and it could be expressed on the HUVECs. [

Objective To observe the effects of co-transfection of proto-oncogene c-myc antisense oligodeoxynucleotides(c-myc-AODN) and tissue-type plasminogen activator(tPA) gene on intimal hyperplasia of auto-transplantion artery in rabbit. Methods The left and right external iliac artery(length 1.0 cm) of rabbits were cross transplanted. The artery grafts and sutures were respectively soaked in Lipofection, c-myc-AODN, pBudCE4.1/tPA, c-myc-AODN and pBudCE4.1/tPA solution for 15 minutes. Each group were divided into five subgroups(n=5, in each subgroup) according to the sacrifice times(3, 7, 14, 28 and 56 d after operation). Specimens were harvested for pathology, chromogenic substrate test, 3H-TdR incorporation test and immunochemisty coloration study. Result The intimal area, stenosis ratio, 3H-TdR incorporation, PCNA positive cell in c-myc-AODN adding tPA co-transfection group were significantly lower than that of control group(P0.01), and that were lower than c-myc-AODN transfection group and tPA gene transfection group(P0.05). Conclusion Vascular local co-transfection of tPA gene and c-myc-AODN effectively inhibits the proliferation of vascular smooth muscle cell(VSMC) and hyperplasia of intima of the transplanted artery.

Animals ; Biological Warfare ; Burns ; pathology ; Burns, Chemical ; pathology ; Chemical Warfare ; China ; Humans ; Military Medicine ; Radiation Injuries ; pathology ; Wounds, Gunshot ; pathology

OBJECTIVETo investigate the effect of oligochitosan in promoting intestinal absorption of protoberberine alkaloids in extracts from Corydalis saxicola total alkaloids.

METHODThe in vitro single-pass intestinal perfusion model in rats was established to study the changes in absorption kinetic parameters of dehydrocavidine, berberine hydrochloride and palmatine chloride in C. saxicola total alkaloids after the addition of different concentrations oligochitosan and evaluate the effect of oligochitosan in promoting intestinal absorption of the drugs.

RESULTThe concentration of oligochitosan had different effects on the absorption rate constant (Ka) and apparent permeability coefficient (Peff) of the three active component in rat intestines. Ka and Peff in 0.5% oligochitosan group significantly increased, indicating a stronger effect in promoting the absorption.

CONCLUSIONOligochitosan has a certain effect in promoting the intestinal absorptions of protoberberine alkaloids in C. saxicola total alkaloids.

Animals ; Berberine Alkaloids ; administration & dosage ; pharmacokinetics ; Chitin ; administration & dosage ; analogs & derivatives ; Corydalis ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Objective To study the mechanism of pregnancy immune tolerance induced by adoptive transferring of FasL gene-modified dendritic cells(DC). Methods The mouse models of spontaneous abortion(CBA/J ? DBA/2) and normal pregnancy(CBA/J ? BALB/c) were established.Five different experimental groups were included: mice of normal pregnancy(CBA/J?BALB/c)(n=17),served as control group;mice of spontaneous abortion without treatment(CBA/J?DBA/2)(n=37),mice injected with DC culture medium(DCCM)(n=25);mice immunized with empty plasmid pcDNA3.1-DC(n=6);and mice immunized with pcDNA3.1-FasL-DC(n=5).Embryo resorption rates of pregnant mice in each groups were observed.Annexin V-FITC was used to detect the apoptosis of T cells.Immunohistochemical staining(SABC) was used to detect the expression of FasL in decidual membranes of pregnant mice. ResultsThe embryo resorption rate of mice immunized with FasL-DC was decreased significantly as compared with that of mice of spontaneous abortion without treatment,DCCM group and immunized with pcDNA3.1-DC(P0.05). Conclusion The decreased apoptosis rate of peripheral T cells and the reduced expression of FasL in decidual membranes may be an important mechanism for the pathogenesis of abortion.Adoptive transfer of FasL gene-modified DC may induce pregnancy immune tolerance by increasing FasL expression of maternal-fetal interface and decreasing embryo resorption rate.

Objective To evaluate diagnostic value of X-ray computed tomography(CT) neck masses in children.Methods Clinic,pathologic diagnosis and CT scans of 26 patients from Jan.2004 to Jun.2006 with neck masses which location,density,edge and near construction were reviewed retrospectively.Results Seven cases in neck anterior area,13 cases in neck lateral area,6 cases in other areas or unconfirmed.Eight cases with lower density than cervical muscles,2 cases with same density,2 cases with higher density,14 cases with mixed density.Inflammatory masses were observed in 13 cases,congenital malformation in 11 cases,thyroiditis in 1 case,pharyngeal tumor-like proliferation in 1 case.Conclusion CT scan is the best choice of diagnosis of neck masses in children,and is valuable to diagnosis associated with history and physical examination.

Crystallization has been widely applied in pharmaceutical formulations as an effective approach to improve the stability and efficacy of small agents. However protein crystals are suffered from limitation in the drug delivery system due to their complex crystallization behaviors. With development of crystallization technologies and their industrial application, protein crystals are receiving more and more attentions as a novel delivery system for biomacromolecules. Crystals with thermodynamic stable structure can improve the physical and chemical stability of protein drugs and present a sustained release behavior. On the basis of pertinent literatures, this review introduces the recent research situation and development process of protein crystals as drug delivery system. Moreover, the crystallization process of proteins, as well as the preparation and potential application are discussed systematically.
Crystallization ; Drug Delivery Systems ; Pharmaceutical Preparations ; administration & dosage ; Proteins ; chemistry

OBJECTIVETo investigate the clinical application value of 8.5/11.5 F transurethral seminal vesiculoscopy in the diagnosis and treatment of refractory hematospermia.

METHODSWe retrospectively analyzed 78 cases of refractory hematospermia diagnosed and treated by 8.5/11.5 F transurethral seminal vesiculoscopy from June 2012 to June 2014. The patients underwent serum PSA examination, transrectal ultrasonography, seminal vesicle ultrasonography, and pelvis CT or MRI before surgery, and all received transurethral seminal vesiculoscopy under the 8.5/11.5 F rigid ureteroscope.

RESULTSOperations were all successfully accomplished, which revealed abnormal opening of the ejaculatory duct in 5 cases, mucosal inflammatory hyperemia in the prostatic utricle and seminal vesicle in 78, dark red mucilage substance in the seminal vesicle in 34, seminal vesicle stones in 19, small polyp in the seminal vesicle in 2, and ejaculatory duct or seminal vesicle cyst in 4. All the patients received symptomatic treatment during the surgery. After surgery, hematouria was found in 13 cases, which disappeared within 2 weeks, pelvic hematoma in 1 case, which was cured by conservative treatment within 3 months, and epididymitis in 2 cases, which was controlled by anti-infection treatment. Hematospermia recurred in 3 cases during the 1-year postoperative follow-up.

CONCLUSION8.5/11.5 F transurethral seminal vesiculoscopy, with its advantages of easy operation, wide field of vision, large channel for operation, and few complications, deserves general clinical application in the diagnosis and treatment of refractory hematospermia.

Calculi ; Ejaculatory Ducts ; Endoscopy ; methods ; Epididymitis ; etiology ; Hemospermia ; diagnosis ; therapy ; Humans ; Magnetic Resonance Imaging ; Male ; Postoperative Period ; Recurrence ; Retrospective Studies ; Seminal Vesicles ; Tomography, X-Ray Computed ; Urethra

OBJECTIVETo investigate approach and possibility of transferring basic fibroblast growth factor (bFGF) gene into rabbit bone marrow stromal cells (BMSCs).

METHODSThe eukaryotic expression vectors harboring bFGF cDNA were constructed and transfected into rabbit BMSCs mediated by liposome. The transcription and expression of bFGF gene in the transfected BMSCs were detected by means of morphological observation, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA) and RT-PCR. The changes in the biological characteristics of the transfected MSCs were also observed.

RESULTSStable overexpression of bFGF protein was detected in the transfected BMSCs, which showed differentiation towards chondrocyte lineage.

CONCLUSIONStable expression of bFGF gene in transfected BMSCs can induce cell differentiation into chondrocyte lineage.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Female ; Fibroblast Growth Factor 2 ; biosynthesis ; genetics ; Gene Expression ; Genetic Vectors ; genetics ; Male ; Rabbits ; Stromal Cells ; cytology ; metabolism ; Transfection

Objective To ascertain the level and dynamic changes of environmental health hazards in the rural areas of Pukou, Nanjing, to objectively evaluate the situation of rural environmental sanitation, providing basis and support for formulating policies and measures by the higher authorities. Methods According to the requirements of the rural environmental sanitation monitoring program of Jiangsu Province in 2017, 5 streets, 20 administrative villages were selected as monitoring points and 100 families used as monitoring objects.The monitoring data were obtained through interviews, access data, field records and laboratory tests. Results Centralized water supply was realized by 20 monitoring points.The rate of collection and centralized disposal of domestic waste was 95.00%.The sewage disposal sites were mainly discharged into sewage treatment plants and harmless sanitary toilets, accounting for 41.00% and 42.00% respectively.The coverage rate of sanitary toilets was 95.54%.Those who did the work of killing rats, flies and mosquitoes accounted for 40.00% and those who did the work of killing cockroaches 25.00%.And 98.00% of the families used fuel for gas, 98.00% of the families never burned straw in the fields, 88.00% of the families did not use the film, and the membrane recycling rate was 91.67%.It was found that 100% of the households drank boiled water, and 90.00% of the respondents developed the habit of washing hands before and after meals.The detection rate of Ascaris eggs was 20.00% in 20 soil samples, and the contents of Pb, CD and Cr were all within the normal range. Conclusion Although some achievements have been made in the rural environmental sanitation in Pukou, the management of pollution of rural sewage, waste and feces should be further strengthened, and the dangerous factors that affect the health of rural residents should be decreased and four pests removal activities actively carried out.