Objective To investigate the effect of small interfering RNA (siRNA) targeting cytoplasmic domain of tissue factor on apoptosis of vascular endothelial cells. Methods Specific siRNA targeting cytoplasmic domain of tissue factor were designed, and synthetic oligos were inserted into plasmid DNA. The siRNA constructs were transfected into human umbilical vascular endothelial cells (HUVEC) with liposome. The HUVEC were transfected with the constructs encoding siRNA Ⅰ, siRNA Ⅱ and pcDNA~(TM)6.2 GW/-miR plasmid separately. The transfected HUVEC were mixed with CD8~+ T lymphocytes. The apoptotic rate of tranfected HUVEC mixed with lymphocytes was analyzed by flow cytometry. Magnetic beads were used to measure PT of the supematant in the mixed lymphocytes culture. Results The siRNA constructs were confirmed by DNA sequence analysis. The apoptotic rate of HUVEC transfected with siRNA Ⅰ and Ⅱ plasmids was decreased significantly as compared with the empty control group (P<0.01). The apoptosis rate of HUVEC transfected with siRNA Ⅰ plasmid was lower than that of HUVEC transfected with siRNA Ⅱ plasmid (P<0.05). APTT of the culture supernatants in the three transfection groups was lower in the control groups (P <0.05), but there was significant difference among the three transfection groups. Conclusion The siRNA targeting cytoplasmic domain of tissue factor were successfully constructed, siRNA can protect HUVEC, and reduce the apoptotic rate of endothelial cells in mixed lymphocyte reaction without influencing the coagulation function.

OBJECTIVETo compare robot-assisted laparoscopic radical prostatectomy (RALRP) with laparoscopic radical prostatectomy (LRP) in the treatment of prostate cancer and investigate the clinical application value of RLRP.

METHODSWe retrospectively analyzed 70 cases of prostate cancer treated by RALRP and another 32 cases treated by LRP. We compared the operation time, intraoperative blood loss and transfusion, catheter-indwelling time, postoperative hospital stay, incisal margin positive rate, biochemical recurrence, and normal postoperative urinary continence and penile erectile function between the two groups of patients.

RESULTSAll the operations were successfully accomplished. RALRP exhibited a significant superiority over LRP in intraoperative blood loss and transfusion, catheter-indwelling time, and postoperative hospital stay, urinary continence and erectile function (P < 0.05).

CONCLUSIONRobot-assisted laparoscopic radical prostatectomy, with its advantages of few postoperative complications and well-preserved urinary continence and penile erectile function, is an effective, safe and minimally invasive surgical option for prostate cancer.

Blood Loss, Surgical ; Humans ; Laparoscopy ; Length of Stay ; Male ; Operative Time ; Penile Erection ; Postoperative Complications ; Postoperative Period ; Prostatectomy ; methods ; Prostatic Neoplasms ; surgery ; Retrospective Studies ; Robotic Surgical Procedures ; methods

Objective To investigate the daily total fluoride intake in relating to the prevalence of skeletal fluorosis in two villages in Jiangsu Province,in order to provide the scientific evidences for the control and prevention of endemic fluorosis.Methods Adults sampled from a high-fluoride Village,Wamiao,and a low-fluoride Village,Xinhuai,were surveyed in this study according to the fluoride concentration in their household shallow well.The average daily total fluoride intake from difierent sources and the skeletal fluorosis were investigated in each subject.Then the subjects from two villages were allocated into five subgroups(＜2.00,2.00～,3.00～,4.00～,≥5.00 mg/d),the relation fluoride intake and prevalence of osteofluorosis was analyzed.Results The prevalence of skeletal fluorosis in Wamiao Village was 31.06%(41/132),but no skeletal fluorosis case(0/35)was found in Xinhuai Village.According to the daily total fluoride intake,subjects with higher daily total fluoride intake tended to associated with a higher prevalence of skeletal fluorosis in a significant dose-response relationship(regression equation:y=2.624-6.855x+3.424x2:r=0.997).The benchmark dose lower limitation of daily total fluoride intake with 95% confidencewas 2.50 mg/d calculated according to this dose-response relationship,the reference dose(RfD)was 2.50 mg/d.In Wamiao Village a significant difference was also found between daily total fluoride intake in 41 subjects[(5.09±1.20)mg/d]with X-ray detectable skeletal fluorosis and in 91 subjects[(3.08±1.12)mg/d]without X-ray detectable skeletal fluorosis(t=-9.32,P＜0.01).Conclusions These findings indicate that the daily total fluoride intake has a significant dose-response relationship with the prevalence of skeletal fluorosis in an endemic fluorosis area associated with high-fluoride drinking water;and the RfD in this study was lower than that in the national standard of"Chinese hygienic standard for daily total fluoride intake(WS/T 87-1996)"(3.50 mg/d).

OBJECTIVETo study the effects of excessive fluoride on type I collagen in rat developmental dentine.

METHODSEighty SD rats, 5 days old, were divided into experimental and control groups, 40 in each group. The experimental group received subcutaneous injection of 0.2% NaF every 4 days (the dose was 2 mg NaF per kg body wt). The same volume of 0.9% NaCl was used in the control. Twenty rats in each group were killed 4 days after the second and the seventh injection respectively. The expression of type I collagen was assayed with immunohistochemical technique.

RESULTS4 out of 20 rats after two injections showed abnormal distribution of type I collagen (dense stain of collagen in the odontoblast, aggregation of collagen in the dentine and disordered arrangement of collagen in the predentine; All 20 rats after seven injections showed abnormal distribution of type I collagen.

CONCLUSIONExcessive fluoride may affect the metabolism of type I collagen in rat developmental dentine.

Animals ; Animals, Newborn ; Collagen Type I ; analysis ; Dentin ; chemistry ; drug effects ; growth & development ; Fluorides ; toxicity ; Immunohistochemistry ; Rats ; Rats, Sprague-Dawley

in view of the emphasis on disease management and clinical research, the authors introduced the evaluation and construction methods of the departments based on disease management. Introduction to the selection process of the evaluation and construction methods of such departments, the authors expounded the construction thought of clinical research evaluation system. This paper analyzed the important role of clinical scientific research in the formation of hospital characteristic advantage and leading position, then suggested that hospitals should take research-type diseases as the basis, promote steady breakthroughs in their medical services and scientific research level, and fully reflect the responsibilities of tertiary hospitals.

Autopsy ; Eye Injuries ; Forensic Medicine ; Humans

OBJECTIVEThe protection rate of inoculation with BCG vaccine is only 50 percent, and most of patients with tuberculosis had a history of BCG vaccine inoculation. Adenosine (ADO) has an immunomodulating effect; it promotes immune reaction by increasing number of macrophage and enhancing phagocytosis. The present study was designed to investigate if combined use of adenosine with BCG enhances the anti-Mycobacterium tuberculosis effect of macrophage in mice.

METHODSFifty BALB/C mice were divided randomly into 3 groups: BCG group (n = 21), BCG plus ADO group (n = 21) and control group (n = 8). The mice in BCG and BCG plus ADO groups were inoculated with 0.1 ml BCG intradermally and the mice in BCG plus ADO group were injected intraperitoneally with ADO 30 mg/(kg.d) for 5 days. The mice in BCG group and control group were injected with NS 0.1 ml/d for 5 days. Six weeks after the last injection, all mice were challenged with intravenous 1 x 10(6) CFU human Mycobacterium tuberculosis virulent strain. After challenging, lung and spleen specimens were taken at the 10th, 20th and 30th days from the mice of BCG and BCG plus ADO groups and at the 30th day from mice in control group. The pathological examinations of lung and spleen sections were performed after HE staining and acid-fast staining, and detection of cell apoptosis was also performed.

RESULTSConsolidation with neutrophil infiltration was found in most of the lung tissue taken at the day 30; there were a lot of tuberculous granulomas and Mycobacterium tuberculosis in the lungs of control group. The alveolar septum in BCG gradually became wide and in interstitium lymphocyte infiltration dominated, and there were less tuberculous granulomas but there were large number of Mycobacterium tuberculosis in the lungs from 10th to 30th days after challenging. The widening of alveolar septum and consolidation of lung tissue in BCG plus ADO group became milder with monocytes infiltration, and there were few tuberculosis granulomas and Mycobacterium tuberculosis in the lungs from 10th to 30th days after challenging.

CONCLUSIONADO could increase the number of monocyte-macrophages and promoted anti-bacterial effects of these cells.

Adenosine ; administration & dosage ; immunology ; Animals ; BCG Vaccine ; administration & dosage ; immunology ; Disease Models, Animal ; Drug Therapy, Combination ; Injections, Intradermal ; Injections, Intraperitoneal ; Macrophages ; drug effects ; immunology ; Mice ; Mice, Inbred BALB C ; Mycobacterium tuberculosis ; immunology ; Neutrophil Infiltration ; drug effects ; Phagocytosis ; drug effects ; Tuberculosis, Pulmonary ; immunology ; prevention & control

To screen differentially expressed genes involved in osteogenic differentiation of human bone marrow stromal cells (BMSCs) at defined stages, subtractive cDNA library was established by means of suppression subtractive hybridization. The BMSCs cultured for 12 and 21 d were used as driver and tester, respectively. A subtract library was successfully constructed and five positive clones were selected from the library. Sequencing analysis and homology comparison showed that the five clones differentially expressed in BMSCs cultured for 21 d were at least 90% homologous with the known genes in human GenBank. It was interestingly found that the osteogenic BMSCs cultured for 21 d differentially expressed decorin and Bax inhibitor 1. RT-PCR was performed to confirm the differentially expressed genes. The results showed that the expression of Bax inhibitor 1 was significantly higher in the cells of 21-day than that of 12-day, while the expression of decorin was only detected in the cells of 21-day.
Apoptosis Regulatory Proteins ; genetics ; metabolism ; Cell Differentiation ; genetics ; Cells, Cultured ; Decorin ; genetics ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation ; Gene Library ; Humans ; Membrane Proteins ; genetics ; metabolism ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology

OBJECTIVETo assess the safety of hyperbaric oxygen in the treatment of radiation-induced hemorrhagic cystitis in patients with prostate cancer, and to investigate its effect on the growth of indolent prostate cancer in vivo.

METHODSThirty severe combined-immunodeficient mice received subcutaneous injection of human prostate cancer LNCaP cells. Then they were randomized to an experimental and a control group and exposed to 20 sessions of hyperbaric oxygen and normobaric air, respectively, followed by a 4-week observation on the growth of the transplanted tumors and analyses of their histopathological features at 28 days, including the volume, microvessel density (CD34), apoptosis markers (p53 and p27 proteins) and the proliferation index (Ki-67) of the LNCaP tumors.

RESULTSOn the 28th day after tumor vaccination, the tumor volume was (120 +/- 7.9) mm3 in the HBO and (122 +/- 8.2) mm3 in the control group; the microvessel density and the expressions of Ki-67, p53 and p27 were 39.3 +/- 5.2, (78.1 +/- 7.6)%, (40.4 +/- 6.2)% and (63.7 +/- 5.1)% in the former, and 36.2 +/- 4.9, (75.3 +/- 8.4)%, (44.2 +/- 5.7)% and (61.5 +/- 5.5)% in the latter. There were no significant differences in all the indexes above between the two groups (P > 0.05).

CONCLUSIONHyperbaric oxygen did not promote the growth of indolent prostate cancer in the murine model, nor did it have any significant effect on the new vessels.

Animals ; Cell Line, Tumor ; Humans ; Hyperbaric Oxygenation ; Male ; Mice ; Mice, SCID ; Prostatic Neoplasms ; Xenograft Model Antitumor Assays

OBJECTIVETo study the expression of MMP-8 in human and rat tooth development.

METHODSImmunohistochemistry was used to detect the localization of MMP-8 protein while in situ hybridization was used to examine the expression of MMP-8 mRNA.

RESULTSThe expression of MMP-8 protein was localized in odontoblast and dentin matrix at the later bell stage in human tooth germ. The dentin was denser close to the pulp cavity. The expression of MMP-8 mRNA was found in very few polarized odontoblast at the early bell stage and all polarized odontoblast at the later bell stage in rat tooth germ.

CONCLUSIONThe results suggested that MMP-8 involved in dentin matrix rebuilding in the process of dentin formation in human and rat dental development.

Animals ; Animals, Newborn ; Dentin ; enzymology ; Embryo, Mammalian ; In Situ Hybridization ; Matrix Metalloproteinase 8 ; biosynthesis ; genetics ; Maxilla ; enzymology ; Odontogenesis ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Tooth Germ ; embryology ; enzymology