OBJECTIVETo probe into regularity of effects of electroacupuncture on focal cerebral infarction.

METHODSFocal cerebral ischemia model was established by middle cerebral artery heat-occlusion (MACO) method. Electroacupuncture was given at Dazhui (GV 14) and Baihui (GV 20). NSS was used for evaluation of neurological impairment, TTC staining method for detection of the infarct volume, and HE staining method for investigation of the pathomorphologic lesion of the brain.

RESULTSMCAO could induce neurologic impairment, cerebral infarction and cerebral pathological lesion in the rat, all of these changes could be cured spontaneously in varying degrees with prolongation of ischemic time, but electroacupuncture could relieve the lesions to a certain extents.

CONCLUSIONElectroacupuncture can improve neurologic deficit impairment, reduce the volume of cerebral infarction and ischemic pathological lesion, early interfere of acupuncture and moxibustion is of very important clinical significance for treatment of ischemic apoplexy.

Animals ; Brain ; Brain Ischemia ; therapy ; Cerebral Infarction ; Electroacupuncture ; Moxibustion ; Rats ; Rats, Sprague-Dawley

Objective To observe the expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblast,and to study the role of fibronectin in pathogenesis of chronic fluorosis.Methods Male and female Wistar rats 144 were randomly divided into four groups,which were designated as the control group(normal diets,n =36),fluoride group(normal diets + 100 mg/L fluoride,n =36),lower calcium monophagia group (synthetic diets,n =36) and lower calcium monOphagia with fluoride group(synthetic diets + 100 mg/L fluoride,n =36).Rats were sacrificed 4 and 8 months after beginning of the experiment,respectively,and femur tissue was fixated and paraffin-embedded.The osteoblast isolated from calvaria of neonatal rats was treated with different dose of fluoride(0,1,2,4 mg/L fluoride,respectively) for 48 and 72 h,cell culture supernatant and cells were collected,respectively.The cranial osteoblasts were cultured in vitro and divided into four groups according to different concentration of fluoride added,which were 0(control group),0.01,1.00,and 10.00 mg/L groups.These cells were treated with mineralized induced medium at day 2 and cultured for 3 more weeks whereafter,and then the slides were fixed in alcohol.The expression of fibronectin in rat femur tissue was detected by immunohistochemistry (IHC),and fibronectin mRNA expression was determined by in situ hybridization; the fibronectin levels in supernatant of cultured osteoblast was examined by enzyme-linked immunosorbent assay(ELISA),and the expression of fibronectin mRNA in osteoblasts was detected with RT-PCR; skull mineralized nodule formation of osteoblasts was observed under a light microscopy after stained with 0.1％ red alizarin liquid.Results Little expression of fibronectin (brown granules under light microscope) could be seen in femur tissue of fluorosis rats of control group and lower calcium monophagia group; but abundantly expressed in fluoride group and lower calcium monophagia with fluoride group; the fibronectin was also expressed in osteoblasts,bone cells and bone marrow cells with less red particles in the control group and lower calcium monophagia group,but more in the fluoride group and lower calcium monophagia with fluoride group.The expression of fibronectin protein in supernatant of cultured osteoblasts was significantly increased in the group of 4 mg/L fluoride at 48 h(0.108 ± 0.042,t =0.764,P＜ 0.05) compared with control group(0.081 ± 0.010); the value was also significantly increased in 1,2,4 mg/L groups at 72 h(0.089 ± 0.010,0.087 ± 0.012,0.098 ± 0.023; t =0.765,0.704,0.996; all P ＜ 0.05) compared with control group (0.070 ± 0.014) ; the expression of fibronectin mRNA was much higher in 1,2,4 mg/L groups at 48 h (0.61 ±0.06,0.77 ± 0.07,0.77 ± 0.07) and 72 h(1.61 ± 0.14,2.54 ± 0.20,2.75 ± 0.22) compared with control group [0.48 ± 0.04(t =0.111,0.182,0.182,all P ＜ 0.05),0.97 ± 0.08(t =0.093,0.109,0.108,all P＜ 0.05) ].A lot of mineralized nodules could be seen under light microscope in 1.00 and 10.00 mg/L groups.Conclusions The expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblasts are increased,and fluoride also promotes the mineralization nodules formation of osteoblasts.These results suggest that fibronectin may regulate the process of bone mineralization,and possibly play a role in the development of skeletal fluorosis.

Objective:To retrospectively analyze the data of patients undergoing extracorporeal membrane oxygenation (ECMO) during perioperative period of cardiac transplantation and provide objective and reliable evidence for further clinical promotion.Methods:Collect the clinical data of patients undergoing heart transplantation and ECMO support in Fuwai Hospital, analyze the duration of ECMO support, combined use of aortic balloon counterpulsation (IABP), and complications during the supporting period. All statistical analyses were processed by SPSS 23.0 software. Independent sample Student's test was employed for normal distribution and Mann-Whitney U test for abormal distribution. χ2 or Fisher exact test was utilized for comparinge the classification data between groups. Results:All ECMO support models were intravenous-arterial ECMO (V-A ECMO). Eight patients successfully bridging heart transplantation through VA-ECMO. Sixty-one patients (89.7%) who had undergone cardiac transplantation were successfully weaned from ECMO while 48 patients (70.5%) survived and discharged. The most common complications during circulation support are bleeding, acute renal insufficiency, and pulmonary infection. Patients with ECMO support in the operating room had a better rate of survival and weaning off(95.6%, 84.4%) than those with ECMO at the bedside of ICU(72.2%, 27.8%).Conclusions:ECMO can provide adequate circulation and respiratory function support for heart transplant patients, and it is an indispensable treatment for patients to go through the perioperative period of heart transplant surgery smoothly. It is advocated to use IABP combined with ECMO in the early stage and at the same time to increase the perfusion of vital organs, improve the prognosis of patients and obtain good outcomes.

Background More efforts have been made in the functional protection of the glaucoma ganglion cells (RGCs) nowadays.As main ingredient,astragalus polysaccharides (APS) enhances neuron regeneration protein expression and promotes peripheral nerve recovery.But whether APS has a protecting effect on RGCs is incompletely clear.Objective The purpose of this study was to evaluate the neuroprotective effect of APS on the RGCs in a rat model of experimental glaucoma.Methods Forty-four SPF SD rats were divided into 4 groups randomly as follows:normal control group,negative control group,low dose APS group and high dose APS group,with 10 rats for each group.APS of 500 mg/kg or 2000 mg/kg (2.5 ml) was administered by gavage feeding once daily for 2 weeks in low dose or high dose of APS group,respectively,and the same volume of normal saline solution was applied instead of APS in the model control group.Two weeks later,aspirate 0.2 ml aqueous followed by methylcellulose injected into the anterior chamber to create the acute ocular hypertension model in the three groups above.No any intervention was performed in the normal control group.The rats were sacrificed on the fifth day after model established to take a retinal section.Ocular hypertension-induced damage was evaluated by regular retina histopathologic examination.Immunolhistochemistry for caspase-3 and TUNEL kits were used to determine the expression of caspase-3 protein in retina and apoptosis rate of RGCs.Retinal cross-sections were analyzed by Image Pro Plus 5.1 software to determine the thickness of various retinal layers and the positive staining cell density in the retinal ganglion cell layer (RGCL).Results On the fifth day after establishment of models,intraocular pressure (IOP) was significantly elevated in the model control group,low dose APS group and high dose APS group in comparison with the normal control group (t=-8.900,-10.700,-11.300,P＜0.01).Retinal morphology was normal in the rats of the normal control group,but in the model control group,rat retina was significantly thickened from severe retinal edema and cell arrangement disorder.Mild retinal abnormality was seen in the low dose APS group;while obvious retina edema was in high dose APS group.The entire retinal thickness,outer nuclear layer thickness and retinal nerve fiber thickness values were lower in the low dose APS group than those of model control group (t =-23.700,-14.770,-11.640,P＜0.01).However,no difference was found in outer nuclear layer thickness and retinal nerve fiber thickness values between high dose APS group and normal control group (t =-0.780,-0.460,P ＞ 0.05).Percentage of positive RGCs for caspase-3 protein and rate of apoptotic RGCs were significantly reduced in low dose APS group compared with model control group (caspase-3:F=87.710,P=0.001;RGCs apoptosis:F=272.840,P＜0.01).Conclusions 500 mg/kg APS can protect retina and RGCs against ocular hypertension-induced damage.The protection of APS is non-dosedependent.

Nephron sparing surgery ( NSS ) has become the standard treatment of small renal cancer . NSS have the similar curative effect compared with radical nephrectomy and preserve the kidney fuction. However, positive surgical margins after NSS is increasing and has attracted more and more attention. We will discuss positive surgical margins related factors and how to reduce the positive surgical margins in this review.

Objective To evaluate the possibility of short interfering RNA (siRNA) inhibiting Coxsackievirns B3 (CVB3) infection in vitro, and discover the mechanism initially. Methods We obtained proper effective dosage of siRNA by observing cytopathic effect (CPE). Estimate its antiviral activities and its pathway of siRNA by Western Blot assay and RT-PCR. Results Results showed that siRNA-3753 can be effectively transfected into HeLa cells, we can achieve a high transfection efficiency up to 98.77 % and its effect can last for 48 h stably in cells. 0.6 μmol/L siRNA-3753 got a high inhibiting effect of virus and didn't show any toxicity to cells. So we consider this concentration as the experimental concentration, siRNA-3753 can debase virus reproduction. The antiviral effect is sequence-specific and is not attributable to either interferon or the interferon response effectors protein kinase R (PKR). Conclusion The data confirmed that siRNA can effectively inhibit CVB3 infection in vitro, its antivirus effect was gained from specific debase of virus genome.

To study the inhibitory effect and function characteristics of small interfering RNA (siRNA) on cosxackievirus B3(CVB3) infection by RNA interference technique, siRNA-2B against the viral 2B region was synthesized and transfected into HeLa cell, which was then infected with CVB3. The efficiency of siRNA transfection was examined by FCM, the cell toxicity of siRNA-2B by MTT, and the antiviral ability of siRNA-2B by cytopathic effect (CPE), plaque reduction assay and RT-PCR. The results showed that siRNA-2B could be transfected efficiently into HeLa cell and lasted at least 48h. High concentration of siRNA-2B didn't show any sign of toxicity to cells. siRNA-2B exhibited a significant protective effect on cell viability by specific inhibition of viral replication. It showed a close relationship between the concentrations of siRNA-2B and the antiviral effects. siRNA-2B led to dramatical reduction of viral titers in supernatant of cell culture and weakened the reinfection ability of the virus. These data proposed that siRNA-2B, targeting 2B protein, can effectively inhibit CVB3 infection in HeLa cell and exhibits its transfection efficiency, viral inhibition specificity and adose-dependant manner, suggesting its potential role in prevention and treatement of CVB3 infection.
Enterovirus ; genetics ; growth & development ; Green Fluorescent Proteins ; genetics ; metabolism ; HeLa Cells ; Humans ; Microscopy, Fluorescence ; Plasmids ; genetics ; RNA, Small Interfering ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Viral Nonstructural Proteins ; genetics ; Virus Replication ; genetics ; physiology

OBJECTIVETo study the cell biocompatibility of porous biphasic calcium phosphate nanocomposite in vitro.

METHODSBone marrow mesenchymal cell (BMSCs) obtained from SD rat bone marrow were in vitro induced and proliferated. Afler their osteoblast phenotypes were verified, BMSCs were seeded onto prepared porous biphasic calcium phosphate nanocomposite (Experiment group) and common porous hydroxyapatite (Control group). The cell adhesion was evaluated by scanning electron microscope. Synthesis of alkaline phosphatase enzyme (ALP) and osteocalcin were detected and cell cycle was detected by flow cytometry.

RESULTSBMSCs could fully attach to and extend on the material in experiment and control group, Moreover, experiment group were superior to control group in adhesion, proliferative abilities and osteogenic activity.

CONCLUSIONBMSCs can differentiate to osteoblast phenotype; the porous biphasic calcium phosphate nanocomposite as bone tissue engineering scaffold has good cell biocompatibility.

Alkaline Phosphatase ; Animals ; Bone Marrow Cells ; Bone and Bones ; Cell Adhesion ; Durapatite ; Hydroxyapatites ; Materials Testing ; Nanocomposites ; Osteoblasts ; Osteocalcin ; Rats ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVETo evaluate feasibility of inhibiting coxsackievirus B3 (CVB3) infection at cellular, protein and gene levels by using small interfering RNA (siRNA).

METHODSAntiviral activities of siRNAs were evaluated by observing cytopathic effect (CPE), using plaque reduction Western blotting assays and RT-PCR.

RESULTSEight siRNAs were synthesized, among them, SiRNA-2, SiRNA-3, SiRNA-6 and SiRNA-7 which were targeted against sequences located in 2B, VP4, 2A and 3C section of CVB3 genome, were designed to have different effect of inhibiting CVB3 infection in vitro. SiRNA-2 showed the best protective effect, 95 percent inhibition of CVB3 cytopathic effect and plaque forming effect was observed at 0.0001 MOI, viral protein synthesis and replication were inhibited. SiRNA-2 showed 30 percent inhibition of virus at 0.1 MOI, 70 percent inhibition at 0.01 MOI, 88 percent inhibition at 0.001 MOI, and 99 percent inhibition at 0.0001 MOI 48 hours after CVB3 infection.

CONCLUSIONSiRNA could effectively inhibit CVB3 infection in vitro, siRNA-2, which is targeted against sequence in 2B section of CVB3 genome, seemed to be the best one among those synthesized in this study.

Coxsackievirus Infections ; therapy ; virology ; Cytopathogenic Effect, Viral ; drug effects ; Enterovirus ; genetics ; physiology ; HeLa Cells ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; therapeutic use ; Virus Replication ; drug effects

BACKGROUNDAdvances in minimally invasive surgical techniques and neonatal intensive care for neonates have allowed for repair of the neonatal esophageal atresia with tracheoesophageal fistula (EA/TEF) to be approached endoscopically. However, thoracoscopic surgery in children is still performed in only a few centers throughout the world. The aim of this study was to compare the neonatal tolerance to the thoracoscopic repair (TR) and the open repair (OR) and also to discuss anesthetic management in thoracoscopic procedure.

METHODSWe performed a prospective study enrolling newborns diagnosed with EA with distal TEF (type C) receiving the repair surgery between June 2009 and January 2012 in our institution. Data collected included the newborns' gestational age and weight at the time of the operation, operative time, parameters of intraoperative mechanical ventilation, oxygenation, end-tidal carbon dioxide (ETCO2), and analysis of blood gases. Time to extubation and length of stay were also recorded.

RESULTSIntravenous induction with muscle paralysis followed by pressure-control ventilation and tracheal intubation regardless of the position of the fistula can be performed uneventfully in EA/TEF newborns with no additional airway anomalies and large, pericarinal fistulas in our experiences. The thoracoscopic approach appeared to take longer than the open approach. During the procedure of repair, hypercarbia and acidosis developed immediately 1 hour after pneumothorax in both groups. CO2 insufflation did have additional influence on the respiratory function of the newborns in the TR group; values of PaCO2 and ETCO2 were higher in the TR group but the difference did not reach statistical significance. By the end of the procedure, values of PaCO2 and ETCO2 returned to the baseline levels while pH did not, but all parameters made no difference in the two groups. Besides, time to extubation was shorter in the TR group.

CONCLUSIONSThoracoscopic repair of EA/TEF is comparable to the open repair, and is believed to be safe and tolerable in selected patients. A wider range of neonates may be acceptable for thoracoscopic EA/TEF repair with increasing surgical experience.

Esophageal Atresia ; surgery ; Female ; Gestational Age ; Humans ; Infant, Newborn ; Male ; Prospective Studies ; Thoracoscopy ; methods ; Tracheoesophageal Fistula ; surgery