Objective To investigate the effects of bone mesenchymal stem cell (BMSC) transplantation on neuronal nuclei (NeuN) and neurogenin 1 (Ngnl) in focal cerebral ischemic rats.Methods A total of 64 healthy adult male Sprague-Dawley rats were randomly divided into normal (N) + phosphate-buffered solution (PBS),middle cerebral occlusion (MCAO)+ PBS,N + BMSC and MCAO + BMSC groups (n =16 in each group).A rat model was induced by the intraluminal suture method.BMSC was cultured in vitro.At 24 h after modeling,brain transplantation was conducted.Magnetic resonance imaging (MRI) was used to detect infarct volume in vivo.NeuN/DAP,Ngnl/DAPIimmunofluorescence double-labeling and Western blot were used to detect the expression of NeuN and Ngnl around ischemic brain tissue.Results On day 14 after transplantation,the T1-and T2-weighted imaging revealed that the cerebral cortex and striatum had abnormal signal areas in the rats of the MCAO group.The infarct volume of the MCAO + BMSC group was significantly less than that of the MCAO + PBS group (32.5％ ± 4.2％ vs.47.9％ ± 7.9％ ; P ＜ 0.01).Immunofluorescence doublelabeling assay showed that the numbers of cells of NeuN+/DAPI+ (976.2 ± 87.5/mm2 vs.1 908.3 ±127.8/mm2; P ＜ 0.01) and Ngn1 +/DAPI + (251.6 ± 23.1/rmm2 vs.285.1 ± 25.2/mm2 ; P ＜ 0.01) of the MCAO + PBS group were significantly less than those of the N + PBS group,but those of NeuN+/DAPI +(1 439.9 ± 101.7/mm2; P ＜ 0.01) and Ngn1 +/DAPI + (356.3 ± 35.6/mm2; P ＜ 0.01) of the MCAO + BMSC group were significantly more than the MCAO + PBS group.Western blot analysis showed that the protein expression levels of NeuN (0.69 ±0.06 vs.0.91 ±0.09; P ＜0.01) and Ngn1 (0.53 ±0.05 vs.0.62 ±0.07;P ＜0.01) of the MCAO +PBS group were significantly lower than those of the N +PBS group,but those of NeuN (0.82 ± 0.07; P ＜ 0.01) and Ngn1 (0.77 ± 0.09; P ＜ 0.01) of the MCAO + BMSC group were significantly higher than the MCAO + PBS group.Conclusions BMSC transplantation may promote the expression of NeuN and Ngn1,and alleviate MCAO caused brain injury.

Objective: To study the examination of coronary sinus (CS) and blood flow by transthoracic echocardiography(TTE) and transesophageal echocardiography(TEE).Methods: Thirty patients with supraventricular tachycardia were studied by TTE and TEE. The CS was visualized using modified 4 chamber view. The position of the probe was optimized until the coronary sinus with its ostium into the right atrium could be visualized. CS flow recordings were performed by TEE with Doppler sample volume placed in the CS within a distance of no more than 10 mm from its ostium. Results: In all patients the angle between the doppler beam and the long axis of the CS was <30°. The CS was fully displayed in 18 patients by TTE and 28 patients by TEE. The length and width of the CS were (16.53±2.57) mm and (4.51±1.30) mm by TTE, (24.11±2.46) mm and (5.06±0.97) mm by TEE.The CS flow was characterized by biphasic flow.Its flow velocity was (39±7.8), (31±6.1) and (21±4.7) cm/s respectively. The CS flow velocity-imeintegral was(43±11.6),(43±13.0),(27±8.2) cm/s. Conclusion: Echocardiography is reliable for detecting CS and its flow. TTE is more feasible for detecting CS and its flow than TEE.

Objective: To explore the blood velocity variety(BFV) of common carotid artery. Methods: BFV was detected with HDI5000 colour doppler ultrasound in 28 healthy volunteers to observe whether there was bifurcation among BFV of each section (about 6.0 cm) of common carotid artery(CCA). CCA was divided into 5 sections (1 cm) from its beginning to bifurcation. BFV was detected in each section. Influences of age, heart rate, blood pressure on BFV were observed. Results: BFV decreased gradually from its beginning to its divergence, the average decreasing value of each section BFV was 5.37 cm/s, and 4.24 cm/s, the difference between maximal and minimum BFV was 17.35 cm/s and 14.32 cm/s in right, left CCA respectively. Maximal BFV increased with age, blood pressure increased and heart rate decreased. But the change of each section BFV was not influenced by age, blood pressure and heart rate. Conclusion: The results indicate that each section BFV is not consistent in CCA, deviation is more obvious when stenosis is valued by internal carotid artery(ICA) BFV/CCA BFV, it should be valued with maximal BFV of ICA.

This study aims to investigate the preparation process and in vitro release behavior of artesunate polylactic acid microspheres, in order to prepare an artesunate polylactic acid (PLA) administration method suitable for hepatic arterial embolization. With PLA as the material and polyvinyl alcohol (PVA) as the emulsifier, O/W emulsion/solvent evaporation method was adopted to prepare artesunate polylactic acid microspheres, and optimize the preparation process. With drug loading capacity, encapsulation efficiency and particle size as indexes, a single factor analysis was made on PLA concentration, PVA concentration, drug loading ratio and stirring velocity. Through an orthogonal experiment, the optimal processing conditions were determined as follows: PLA concentration was 9. 0% , PVA concentration was 0. 9% , drug loading ratio was 1:2 and stirring velocity was 1 000 r x min(-1). According to the verification of the optimal process, microsphere size, drug loading and entrapment rate of artesunate polylactic acid microspheres were (101.7 +/- 0.37) microm, (30.8 +/- 0.84)%, (53.6 +/- 0.62)%, respectively. The results showed that the optimal process was so reasonable and stable that it could lay foundation for further studies.
Artemisinins ; chemistry ; Calibration ; Drug Compounding ; methods ; Lactic Acid ; chemistry ; Microspheres ; Polyesters ; Polymers ; chemistry ; Polyvinyl Alcohol ; chemistry

BACKGROUND: Bone marrow stromal cells(BMSCs) are the ideal gene target cells and will have a bright future in the gene therapy of spinal cord injury.OBJECTIVE :To detect the expression of glial cell line - derived neurotrophic factor(GDNF) gene after BMSCs were infected by adenovirus-medialed GDNF (Adv-GDNF) in vitro and to explore its biological activity.DESIGN: A randomized controlled trial study.SETTING: Laboratory of Orthopedic DepartmentMATERIALS: The experiment was completed in the Laboratory of Orthopedic Department, Affiliated Tongji Hospital of Tong ji Meidcal College,Huazhong University of Science and Technology. Twenty-four SD rats of either gender, weighing (180 ± 20) g.INTERVENTIONS: BMSCs were infected by Adv-GDNF in vitro and then cocultured with spinal cord dorsal root ganglion. The three methods, immunofluorescent chemistry, reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay(ELISA) were used to evaluate GDNF expression in the BMSCs. The biological activity of GDNF was observed by a phase contrast microscope.MAIN OUTCOME MEASURES:Primary outcomes:①RT-PCR;②results of immunofluorescent chemical examination;③biological activity of GDNF in vitro. Secondary outcomes:①culturing and identification of BMSCs②time-effect relationship of GDNF expression revealed by ELISA.RESULTS: Immunofluorescence displayed expression of GDNF in BMSCs 48hours after Adv-GNDF infection. RT-PCR analysis demonstrated expression of GDNF mRNA 24 hours after Adv-GNDF infection. ELISA confirmed the presence of GDNF in the liquid supernatant of BMSCs 24 hours after Adv-GDNF infectionn and showed that GDNF was secreted. The supernatant can promote the neurite outgrowth in the rat dorsal root ganglion(DRG).CONCLUSION: It is demonstrated that BMSCs infected by Adv-GDNF can express GDNF steadily and the expressed GDNF has the activity of promoting neurite outgrowth, which lays a foundation of the GDNF gene therapy for spinal cord injury.

BACKGROUND: H-type vessels are mainly distributed in the metaphysis, which can promote the proliferation of osteocytes, further accelerating osteogenesis.OBJECTIVE: To investigate the expression of H-type vessels in the subchondral bone during the occurrence and development of osteoarthritis.METHODS: 8-week-old C57 mice were randomly divided into experimental and sham groups, followed by the right medial menisectomy to establish the osteoarthritis models or only articular capsulotomy. The knee samples were removed at 4 weeks postoperatively, and were stained with safranin-O-fast green to evaluate the degree of injury. The expression levels of CD31, Emcn and matrix metalloproteinase 13 were detected by immunofluorescent staining. The changes of the subchondral bone were observed by hematoxylin-eosin staining and the changes of bone mass in the subchondral bone were analyzed by micro-CT.RESULTS AND CONCLUSION: Compared with the sham group, the Osteoarthritis Research Society International scores, expression levels of CD31, Emcn, H-type vessels and matrix metalloproteinase 13, as well as the bone mass in the subchondral bone were significantly increased in the experimental group (P < 0.05). To conclude, the increased H-type vessels in the subchondral bone promote the hyperplasia and remodeling of subchondral bone in the progression of osteoarthritis.

OBJECTIVE:To investigate clinical efficacy of cerebroventricular perfusion and intrathecal perfusion of vancomy-cin assisting with continuous drainage in the treatment of intracranial infection secondary to traumatic brain injury,and its effects on cerebrospinal fluid indexes and intracranial pressure. METHODS:One hundred and eighty patients with intracranial infection secondary to traumatic brain injury selected from Sanya Hospital of TCM during Jan. 2012 to Jun. 2015 were randomly divided into control group and observation group according to lottery,with 90 cases in each group. They were given cerebroventricular perfu-sion and intrathecal perfusion of vancomycin(20 mg dissolved in 5 mL normal saline)in cella lateralis and lumbar cisterna respec-tively combined with continuous drainage,q12 h. Both groups received treatment for 7 d.Clinical efficacy,the time of infection con-trol were compared between 2 groups as well as body temperature,intracranial pressure and cerebrospinal fluid indexes before and after treatment,and the occurrence of ADR. RESULTS:After treatment,total response rate of observation group (95.56%) was significantly higher than that of control group (77.78%),with statistical significance (P<0.05). The time of infection control in observation group [(9.67 ± 1.10)d] was significantly shorter than in control group [(11.84 ± 1.29)d],with statistical significance (P<0.05). Body temperature,intracranial pressure,cerebrospinal fluid protein and leukocyte of 2 groups were significantly lower than before treatment;cerebrospinal fluid glucose level was increased significantly compared to before treatment;above indexes of observation group were significantly better than those of control group,with statistical significance(P<0.05).There was no statisti-cal significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:The intrathecal perfusion of vancomycin as-sisting with continuous drainage in the treatment of intracranial infection secondary to traumatic brain injury can effectively speed up the rehabilitation process,reduce the body temperature and intracranial pressure,and is helpful to improve the relevant cerebro-spinal fluid indexes. Therapeutic efficacy of it is better than that of cerebroventricular perfusion.

Objective To investigate the processing characteristics of locative prepositions in patients with Chinese aphasia,and to provide the theoretical evidence for the rehabilitation of aphasia.Methods Twenty aphasic patients caused by left-hemisphere stroke and twenty matched normal controls were studied.Using the locative prepo- sition repeating task(single words,locative preposition phrases and words in sentences),the comprehension task, filling-gap task,the visual-spatial function task and the short-term memory task,we compared the performance be- tween these two groups.Results The aphasic patients had more difficulty in repeating locative prepositions in sen- tences,in comprehension task and filling-gap task,their short term memory was impaired.Both groups did well in re- peating single words and phrases.Conclusion The processing of locative prepositions was impaired in Chinese aphasics.The repetition of locative prepositions was more difficult than that of phrases and single words.The preposi- tions were often omitted.It might be due to the impairment of their short-term memory,or it might have something to do with role they played in the syntactic structure.The latter might also impact the comprehension and filling-gap score.We should make plans before rehabilitation therapy.

Objective To study the feasibility and long-term effects in 173 infants with congenital ventricular septal defect who underwent transcatheter therapy by double-disk ventricular septal defect occluder. Methods We analyzed the clinical data of 173 infants with ventricular septal defect who were performed interventional therapy followed by a follow up study from December 2002 to October 2013. Results The procedure was performed in 173 infants (male = 81 cases and female = 92 cases) aged 11 month-3 years[(2.1±0.7)years]. The weight were 6-15 kg[(10.2±3.6)kg]. The diameter of the defects ranged from 2.5-9.0 mm[(5.1±1.7) mm]. The characteristics of septal were classiifed into four types:the simply perimembranous ventricular septal defects (88 patient,50.8%), the perimembranous ventricular septal defects with pseudoaneurysm (52 patients, 30.0%), the perimembranous ventricular septal defects complicated with aortic valve prolapse (13 patients, 7.5%), and the intracristal ventricular septal defect (20 patients, 11.7%). The diameter of the occluders were between 4-12 mm[(6.3±2.2)mm]. The period of follow-up ranged from 1 month-10 years[(6.2±1.3)years]. The 168 defects were completely occluded in 173 patients(97.1%closure rate) except 5 infants, which 3 patients because the occluder hinder the function of aortic valves and the other occurred complete atrioventricular block(CAVB) when the catheter through defect. One patient occurred CAVB on the third day after the procedure, was reversed by dexamethasone after 4 days. 3 patients with complete left bundle branch block(CLBBB) after the procedure and one went to sustained, but did not observed heart failure in this case during 5 years follow-up. One patient observed sustained CLBBB on the fourth year after the procedure, also did not observed heart failure in this case during 3 years follow-up. 8 patients with trivial residual shun and 4 patients with newly appearance of trivial aortic regurgitation after the procedure, the shun all disappear at one year and the regurgitation did not progress during longest follow-up period at 8.5 years. No other complications, including late-onset CAVB, hematolysis, the occluder displacement and detachment, occurred during 1 month-10 years of follow-up. Conclusions Transcatheter closure of congenital ventricular septal defects is an efifcient method that can be safely used in the majority of infants with ventricular septal defects and have signiifcant long-term effects.