ObjectiveTo compare the effects of normal saline (NS), 10% hydroxyethyl starch (HES) and hypertonic-hyperoncotic solution (HHS,7.5% NaCl/10% HES) on regional cerebral blood flow (rCBF), brain edema and blood-brain barrier (BBB) in a rat model of traumatic head injury (THI) complicate with hemorrhagic shock. Methods60 SD rats were randomized into 5 groups: sham group (n=12), model group (n=12), NS group (n=12), HES group (n=12) and HHS group (n=12). rCBF and mean arterial pressure (MAP) were determined before and after THI, hemorrhagic shock and resuscitation. Cerebral water content and Evans Blue (EB) content were assessed 3 h after resuscitation.ResultsMAP and rCBF were restored to baseline values immediately after resuscitation in all resuscitated group and began to decrease 15 min, 30 min or 45 min later, respectively(P<0.05). 3 h after resuscitation, the brain water content was higher in NS group than those in sham or HHS group(P<0.05). EB contents of injured hemispheres were higher in model and NS group than those in HES or HHS group(P<0.05). ConclusionSmall-volume resuscitation with HHS can restore MAP and rCBF, decrease brain edema and improve BBB in a rat model of THI complicate with hemorrhagic shock.

Objective To assess the value of bulbocavernosus reflex (BCR) in the differential diagnosis of Parkinson' s disease (PD) and multiple system atrophy (MSA).Methods BCR was compared in 55 patients with PD,20 patients with MSA,and 50 healthy controls,who were enrolled from the Department of Neurology,the First Affiliated Hospital of Wenzhou Medical University from May 2013 to April 2014.The clinical features of autonomic nerves dysfunction were also recorded.Results Among all related autonomic symptoms,the occurrence rate of constipation,urinary urgency and frequency in patients with MSA was higher than those with PD.The elicit percent of BCR in patients with PD was 93％,higher than those with MSA (70％).The average latency of BCR in patients with MSA was longer than those with PD (tmale left =16.275,tmale right =14.269,tfemale left =5.954,tfemmale right =5.905,all P ＜ 0.05).The degree of BCR amplitude decreasing in three groups was MSA ＞ PD ＞ healthy controls.There was statistically significant difference among three groups (Fmale left =75.73,Fmale right =73.13,Ffemale left =72.70,Ffemale right =59.44,all P ＜ 0.05).The area under receiver operating characteristic curve (ROC) in differential diagnosis of PD and MSA of the average latency of BCR in male and female was 0.947 and 0.948.The area under ROC curve in differential diagnosis of PD and MSA of the average amplitude of BCR in male and female was 0.886 and 0.920.The ROC curve showed the average latency of BCR in male of 44.80 ms with a sensitivity of 95％ and a specificity of 84％,and in female of 61.35 ms with a sensitivity of 86％ and a specificity of 88％ ; the average amplitude of BCR in male of 0.37 mV with a sensitivity of 96％ and a specificity of 68％,and in female of 0.36 mV with a sensitivity of 98％ and a specificity of 76％,which were critical cutoff values in differential diagnosis of PD and MSA with the best sensitivity and specificity.Conclusion The latency and amplitude of BCR test helps to increase the accuracy in the differential diagnosis of PD and MSA.

Objective To investigate the effect of green light laser in complex posterior urethral stricture after surgical treatment of benign prostatic hyperplasia (BPH).Methods Green light laser was applied in treating 20 cases of complex iatrogenic posterior urethral stricture.Of these cases,12 had false passages,5 had more than 2 strictures and 5 had concurrently urethratresia.The scar tissues were transure- thrally vaporized and resected.The in-dwelling urethral catheter time was 1-2 months after operation. Results All the patients were initially cured without serious complications.The mean operative time was 39 rain (range,30-65 min).During the follow-up of 2-10 months,1 case had mild incontinence:another case (Q_(max)＜9ml/s 2 weeks after surgery) got satisfactory results(Q_(max)＞15ml/s)after the scheduled urethral dilatation.The other 18 cases were treated successfully and voided fluently with postoperative Q_(max)＞15ml/s in all.Conclusions It is suggested that transurethral green light laser procedure is not only safe and ef- fective,but also simple and minimally iuvasive for complex posterior urethral stricture following surgical treat- ment of BPH.

Accidents, Occupational ; statistics & numerical data ; China ; epidemiology ; Humans

OBJECTIVETo investigate the effects on the phenotype, especially the mineralization ability of human embryonic lung fibroblasts(HELFs) by transfection with adenovirus vector ecoding human bone morphogenetic protein-2 gene (Ad-BMP-2).

METHODSThe HELFs were primarily cultured, then transfected with Ad-BMP-2. The morphologic characteristics of the cells were observed. The cell proliferation, alkaline phosphatase activities, BMP-2 protein expression, and the mineralization ability were detected with the methods of MTT, ALP staining, Western blot, and alizarin red S staining, respectively.

RESULTSAfter transfection, the shape of HELFs changed from silm spindle to multifigure, the cells became bigger than before. The colonies changed from unilaminar into multilaminar. The proliferation of HELFs was severely inhibited after transfection. An obvious BMP-2 lane was shown in Western blotting. Most cells presented positive in ALP staining, and large number of nacarat mineralized nodes were observed after alizarin red S staining.

CONCLUSIONHELFs were capable of transforming into osteoblast-like phenotype, and were endowed with the ability of mineralization while being transfected with Ad-BMP-2.

Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; Cell Proliferation ; Fibroblasts ; Genetic Vectors ; Humans ; Osteoblasts ; Osteogenesis ; Phenotype ; Transfection ; Transforming Growth Factor beta

OBJECTIVETo establish a flow cytometric method to detect the alteration of phenotypes and concentration of circulating microvesicles (MVs) from myocardial ischemic preconditioning (IPC) treated rats (IPC-MVs), and to investigate the effects of IPC-MVs on ischemia/reperfusion (I/R) injury in rats.

METHODSMyocardial IPC was elicited by three.cycles of 5-min ischemia and 5-min reperfusion of the left anterior descending (LAD) coronary artery. Platelet-free plasma (PFP) was isolated through two steps of centrifugation at room temperature from the peripheral blood, and IPC-MVs were isolated by ultracentrifugation from PFR PFP was incubated with anti-CD61, anti-CD144, anti-CD45 and anti-Erythroid Cells, and added 1, 2 µm latex beads to calibrate and absolutely count by flow cytometry. For functional research, I/R injury was induced by 30-min ischemia and 120-min reperfusion of LAD. IPC-MVs 7 mg/kg were infused via the femoral vein in myocardial I/R injured rats. Mean arterial blood pressure (MAP), heart rate (HR) and ST-segment of electro-cardiogram (ECG) were monitored throughout the experiment. Changes of myocardial morphology were observed after hematoxylin-eosin (HE) staining. The activity of plasma lactate dehydrogenase (LDH) was tested by Microplate Reader. Myocardial infarct size was measured by TTC staining.

RESULTSTotal IPC-MVs and different phenotypes, including platelet-derived MVs (PMVs), endothelial cell-derived MVs (EMVs), leucocyte-derived MVs (LMVs) and erythrocyte-derived MVs (RMVs) were all isolated which were identified membrane vesicles (<1 Vm) with corresponding antibody positive. The numbers of PMVs, EMVs and RMVs were significantly increased in circulation of IPC treated rats (P<0.05, respectively). In addition, at the end of 120-min reperfusion in I/R injured rats, IPC-MVs markedly increased HR (P<0.01), decreased ST-segment and LDH activity (P < 0.05, P < 0.01). The damage of myocardium was obviously alleviated and myocardial infarct size was significantly lowered after IPC-MVs treatment (P < 0.01).

CONCLUSIONThe method of flow cytometry was successfully established to detect the phenotypes and concentration alteration of IPC-MVs, including PMVs, EMVs, LMVs and RMVs. Furthermore, circulating IPC-MVs protected myocardium against I/R injury in rats.

Animals ; Cell-Derived Microparticles ; metabolism ; Coronary Vessels ; pathology ; Flow Cytometry ; Heart Rate ; Ischemic Preconditioning, Myocardial ; Myocardial Infarction ; physiopathology ; Myocardial Reperfusion Injury ; physiopathology ; Myocardium ; pathology ; Phenotype ; Rats

Ureteral fibroepithelial polyp accompanied by intussusception is a rare occurrence. Currently, most ureteral polyps could be removed readily by ureteroscopy. Nevertheless, endoscopic resection can be difficult in patient with a large polyp, especially accompanied by an intussusception. We described our experience and laparoscopic technique for treatment of a symptomatic 63-year-old woman who presented with a pedunculated, 9-cm-long, left lower ureteral, fibroepithelial polyp accompanied by a 2-cm-long intussusception.
Female ; Humans ; Intussusception ; pathology ; surgery ; Laparoscopy ; methods ; Middle Aged ; Polyps ; pathology ; surgery ; Ureter ; pathology ; surgery

BACKGROUNDTo investigate the incidence and risk factors of retinopathy of prematurity (ROP) in two Neonatal Intensive Care Units in North and South of China, respectively.

METHODSWe studied data concerning 472 infants with gestational age (GA) ≤ 34 weeks or birth weight (BW) ≤ 2000 g who were admitted to the Zhujiang Hospital of Southern Medical University and the Fourth Hospital of Shijiazhuang between January 1, 2011 and December 31, 2011. Clinical information about perinatal neonates was collected and was confirmed by reviewing medical charts. The incidence and severity of ROP were assessed in the screened population. Main outcome measures are the incidence and severity of ROP. The relationship of clinical risk factors and the development of ROP were analyzed.

RESULTSThe overall incidence of ROP was 12.7%, and the overall incidence of type 1 ROP was 2.3%; 9.4% of infants in Zhujiang Hospital had ROP compared to 15.0% infants in the Fourth Hospital of Shijiazhuang developed ROP, and the difference is statistically significant. ROP was significantly associated with GA (odds ratio [OR]: 0.77 [0.62-0.95], P = 0.015), BW (OR: 0.998 [0.996-0.999], P = 0.008), maternal supplemental oxygen administration before and during delivery (OR: 4.27 [1.21-15.10], P = 0.024) and preeclampsia (OR: 6.07 [1.73-21.36] P = 0.005). The risk factors for ROP are different in two hospitals. In Zhujiang Hospital, BW is the independent risk factors for ROP while GA, BW and preeclampsia in the Fourth Hospital in Shijiazhuang Conclusions: Retinopathy of prematurity incidence is different based on area. Incidence of ROP is still high in China. More efforts need to prevent ROP.

China ; Female ; Humans ; Infant, Newborn ; Intensive Care Units, Neonatal ; statistics & numerical data ; Male ; Retinopathy of Prematurity ; epidemiology ; Risk Factors

OBJECTIVETo investigate the effects of microvesicles (MVs) derived from hypoxia/reoxygenation (H/R)-treated human umbilical vein endothelial cells (HUVECs) on endothelium-dependent relaxation of rat thoracic aortic rings.

METHODSH/R injury model was established to induce HUVECs to release H/R-EMVs. H/R-EMVs from HUVECs were isolated by ultracentrifugation from the conditioned culture medium. H/R-EMVs were characterized using 1 μm latex beads and anti-PE-CD144 by flow cytometry. Thoracic aortic rings of rats were incubated with 2.5, 5, 10, 20 μg/ml H/R-EMVs derived from H/R-treated HUVECs for 4 hours, and their endothelium-dependent relaxation in response to acetylcholine (ACh) or endothelium-independent relaxation in response to sodium nitroprusside (SNP) was recorded in vitro. The nitric oxide (NO) production of ACh-treated thoracic aortic rings of rats was measured using Griess reagent. The expression of endothelial NO synthase (eNOS) and phosphorylated eNOS (p-eNOS, Ser-1177) in the thoracic aortic rings of rats was detected by Western blotting. Furthermore, the levels of SOD and MDA in H/R-EMVs-treated thoracic aortic rings of rats were measured using SOD and MDA kit.

RESULTSH/R-EMVs were induced by H/R-treated HUVECs and isolated by ultracentrifugation. The membrane vesicles (< 1 μm) induced by H/R were CD144 positive. ACh-induced relaxation and NO production of rat thoracic aortic rings were impaired by H/R-EMVs treatment in a concentration-dependent manner (P < 0.05, P < 0.01). The expression of total eNOS (t-eNOS) was not affected by H/R-EMVs. However, the expression of p-eNOS decreased after treated with H/R-EMVs. The activity of SOD decreased and the level of MDA increased in H/R-EMVs treated rat thoracic aortic rings (P < 0.01).

CONCLUSIONACh induced endothelium-dependent relaxation of thoracic aortic rings of rats was impaired by H/R-EMVs in a concentration-dependent manner. The mechanisms included a decrease in NO production, p-eNOS expression and an increase in oxidative stress.

Acetylcholine ; pharmacology ; Animals ; Aorta, Thoracic ; physiology ; Cell Hypoxia ; Endothelium, Vascular ; physiology ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; In Vitro Techniques ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Nitroprusside ; pharmacology ; Oxidative Stress ; Rats

It has been reported that baculoviruses could serve as a new gene-transfer vehicle for mammalian cells. We have previously constructed recombinant baculovirus BacV-CMV-EGFPA and have proven that mammalian cells could be effectively infected by the recombinant baculovirus. In this report, we studied the efficiency of baculovirus to deliver exogenous gene into twenty mammalian cells, including twelve human cell lines (WI-38, Hela, HepG2, 293, PLC/PRF/5, 143B, MCF-7, BGC-223, DMS 114, CNE, Raji, LCL-cm), seven murine cell lines (BNL 1ME A.7R.1, CHO-K1, L-929, JC, PT67, NIH3T3, P815) and one monkey cell line (CV1). Results showed that most mammalian cell lines could be transduced by the recombinant baculovirus, the transduction efficiencies of the human and monkey cell lines were markedly higher than that of murine cell lines, and the transduction efficiencies in adherent culture cell lines higher than that of suspend culture cell lines, implying that the infection efficiency of the baculovirus may be correlative with the organism used and the growth properties of the cell lines. The plasmid pcDNA3. 1-EGFP, which contains the CMV promoter and EGFP reporter gene, was next transfected by LipofectAMINE into a number of mammalian cells, especially those cells that were low in the baculovirus transfection. Results showed that the CMV promoter could effectively direct the expression of the reporter gene in these mammalian cells. Therefore the gene-expression efficiencies in different mammalian cell lines by the recombinant baculovirus which contains the same CMV promoter were dictated by the ability of the baculovirus to enter the cell lines. This study suggested that the recombinant baculovirus vector is more suitable for gene expression in primate adherent culture cells than in murine cells and suspend culture cells.
Animals ; Baculoviridae ; genetics ; Cytomegalovirus ; genetics ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Humans ; Promoter Regions, Genetic ; Spodoptera