[Objeetive] To develop the recombinant human bone morphogenetic protein-2(rhBMP-2)loaded amorphous calcium phosphate(ACP)delayed release nano-sized material,investigate its cytotoxicity of cell,and provide a reference for the experiment of composite material in vivo.[Method]The rhBMP-2/ACP delayed release nano-sized material were prepared by chemical wet method and cultured on rabbit bone marrow-derived mesenchymal stem cells(BMSCs)in vitro.Then the adhesion,proliferation,growth and functional expression of BMSCs were measured.[Result]Cytotoxicity test demonstrated that rhBMP-2/ACP delayed release nano-sized material had not affect the percentage of cell's proliferation with material-extracted liquid cultured with BMSCs and the cytotoxicity was graded zero.The adhesion,proliferation,configuration of the cells on the surface of this material were identical to the control group.[Conclusion]It was suggested that rhBMP-2/ACP delayed release nano-sized material might have good cellular biocompatibility,no cytotoxicity and not effected the normal functional expression of BMSCs in vitro.

[Objective]To study the biocompatibility and security of the recombinant human bone morphogenetic protein-2(rhBMP-2) loaded amorphous calcium phosphate(ACP) delayed release nano-sized material and investigate the feasibility of the clinical use as a kind of bone substitute in bone engineering.[Method]The in vitro hemolyzation,cytotoxicity,microkernel test in marrow smear,acute systemic toxicity,pyrogenicity,subcuticular stimulation reaction and short-term intramuscular implantation,as well as endosseous implantation were performed on the rhBMP-2/ACP delayed release nano-sized material.[Result]The material-extracted liquid induced no hemolyzation,no toxic effects of genetic,no pyrogenic reaction in rabbits,no cytotoxicity cultured in rabbit bone marrow-derived mesenchymal stem cells(BMSCs) in vitro and no acute toxicity in mice.The intramuscular implantation and endosseous implantation in rabbits induced no inflammatory reaction,tissue necrosis and the material was degraded gradually,fused organically with tissues.[Conclusion]The biocompatibility and security of the rhBMP-2/ACP delayed release nano-sized material could meet the requirements given in biological standards for implanted biomaterials ISO10993 and GB/T16886,suggesting that it could be a good bone substitution for the clinical trial.

Objective To compare the effects of 0.9% NaCl (normal saline, NS), 10% hydroxyethyl starch (HES, 200/0.5) and hypertonic-hyperoncotic solution (HHS, 7.5% NaCl-10% HES 1:1) on cerebral extracellular excitatory amino acids (EAA) and inhibitory amino acids (IAA) in a rat model of traumatic head injury (THI) combined with acute hemorrhagic shock (AHS). Methods Nineteen healthy adult male SD rats weighing 300-350 g were randomized into 3 groups: NS group (n = 7); HES group ( n = 6) and HHS group ( n = 6). THI was produced by modified Feeney method (a 20g weight drops from a height of 40 cm on the parietal region) and AHS was induced by modified Wiggers method (Blood was removed from femoral artery and MAP was maintained at 40 mm Hg for 1 hour). Fluid resuscitation was started at 1 hour of AHS with 0.9% NS (3 times the volume of the removed whole blood) or 10% HES (in a one to one ratio) or HHS 4 ml?kg-1 administered over 15 min. The extracellular fluid of injured brain tissue was collected using intracerebral microdialysis before head injury (baseline) during THI + AHS (1h) and resuscitatin (2h) for determination of the levels of EAA (glutamate, aspartate) and IAA (glycine, GABA, taurine) by HPLC.Results The 5 amino acids were significantly increased during THI + AHS as compared with their baseline values. Glutamate level was further increased during resuscitation with NS. GABA and taurine concentrations were maintained at high level during resuscitation with HES or HHS. The increase in glutamate was inhibited by resuscitation with HES and the increase in glutamate, aspartate and glycine were inhibited by HHS. Conclusions In a rat model of THI combined with AHS, resuscitation with NS can increase cerebral extracellular EAA. Both HES and HHS resuscitation can inhibit the increase in cerebral extracellulaar EAA and HHS is more effective.

Background:Intestinal permeability plays an important role in the development of ulcerative colitis. Sugar molecular probes is a safe and non-invasive method to measure intestinal permeability,and its correlation with inflammatory factors and claudin-8 is not clear. Aims:To explore the correlation of sugar molecular probes with TNF-α,CRP and claudin-8 in intestinal permeability in colitis rats. Methods:Twenty-four rats were assigned randomly to model group and normal control group. Colitis model was induced by DSS solution. Expressions of TNF-α,CRP were determined by ELISA,and expression of claudin-8 was determined by immunohistochemistry. Sugar molecular probes were determined by high performance liquid chromatography. Correlations of inflammatory factors and claudin-8 with sugar molecular probes were analyzed. Results:Compared with normal control group,expressions of colonic tissue TNF-α and CRP were significantly increased while expression of claudin-8 was significantly decreased in model group(P < 0. 01);secretion of lactulose and sucrolose,ratio of lactulose/ mannitol( L/ M)were significantly increased( P < 0. 01)while mannitol secretion was significantly decreased(P < 0. 01). Secretion of lactulose and sucrolose were positively correlated with expressions of TNF-α and CRP(P < 0. 01),but negatively with expression of claudin-8(P < 0. 01). L/ M ratio and mannitol secretion were negatively correlated with expressions of TNF-α and CRP(P < 0. 01),but positively with expression of claudin-8(P <0. 01). Conclusions:Sugar molecular probes and expressions of TNF-α,CRP,claudin-8 have similar results in predicting intestinal permeability in rats. Sugar molecular probes can be used as a potential method to measure intestinal permeability.

BACKGROUND:Cerebrovascular drag, occlusion and other mechanical stimulations inevitably occur during some craniocerebral operations, which cause acute mechanical cerebrovascular vasospasm. At present, the mechanism underlying the patho-physiology as well as the pathological prognosis of this acute mechanical vasospasm remains unclear.OBJECTIVE: To observe changes in the vascular diameter of the middle cerebral artery, cerebral blood flow (CBF), and ultrastructure of vascular wall and vascular endothelial cells, during the early stage (2 hours) of mechanical cerebral vasospasm in cats.DESIGN: Open experiment.SETTING: Department of Neurology, Beijing Tiantan Hospital, Capital Medical University; Beijing Institute of Neurosurgery.MATERIALS: Six healthy adult hybrid cats, of either gender, weighing from 2.5 to 3.5 kg, were provided by the China Medical Science Institute of Experimental Animals. Laser Doppler flowmetry (Periflux 5010, Sweden Perimed Company)was used.METHODS: This study was carried out in the Beijing Institute of Neurosurgery between August 2005 and March 2006. For all experimental surgical procedures, the cats were anesthetized by intraperitoneal injection of 200 g/L chloral hydrate, at 2 mL/kg, and then placed in a prone position. A median incision was made in the scalp and a square bone window, 8×10 mm, was opened at 1.5 cm posterior and 1.5 lateral to the anterior fontanel, after which the dura mater was pricked out. The fine detecting head of the Laser Doppler flowmetry was fixed to a region of the cerebral surfacewith no vessels or with only a few vessels. Subsequently, the cats were placed in lateral position. Under the surgical microscope, the right middle cerebral artery was exposed through a suborbital approach. Blunt apparatus was used to stimulate middle the middle cerebral artery repeatedly, at a frequency of 100 time/min within 30 minutes.The diameter of the middle cerebral artery was measured and a perfusion index of cortical brain tissue was monitored, separately, before and then at 0, 0.5, 1.0, 1.5, 2.0 hours after stimulation. Ultrastructural changes in the vascular wall and the vascular endothelial cells were observed during the early stage (2 hours) of mechanical cerebral vasospasm in cats.MAIN OUTCOME MEASURES: Diameter of the middle cerebral artery and the perfusion index of cortical brain tissue before and then at 0, 0.5, 1.0, 1.5, 2.0 hours after stimulation, as well as any ultrastructural changes in the vascular wall and endothelial cells 2 hours after stimulation.RESULTS: The results from six cats were were analyzed. ①The diameter of the middle cerebral artery was (0.617±0.129), (0.723±0.082), (0.840±0.084) mm 0, 0.5 and 1.0 hours after stimulation, respectively, which was significantly smaller than that before stimulation [(0.897±0.066) mm,t =4.74, 4.017, 1.299,P ＜ 0.01]. ② The perfusion index of cortical brain tissue was 67.8±18.5, 82.5±17.5, 89.8±24.0, 94.0±22.2 and 98.5±21.0 at 0, 0.5, 1.0,1.5 and 2.0 hours after stimulation, which was significantly lower than that before stimulation (159.2±23.5, t =4.716-7.469, P ＜ 0.01 ). ③ At the early stage of acute mechanical stimulation (2 hours) to middle cerebral artery, endothelial cell chromatin aggregated at the edge of the cells and achromocyte formed, but mitochondrial crista was unclear.CONCLUSION: Mechanical stimulation to the middle cerebral artery in cats can lead to cerebral vasospasm. Apoptosis of endothelial cells appears at the early stage of stimulation (2 hours). These results indicate that, in order to prevent against cerebral vasospasm, cerebrovascular mechanical stimulation should be as minimal as possible and that as few as possible craniocerebral operations should be performed.

Objective To discuss the relation between gynecologic tumor and dysmenorrheal and tongue petechia, make clear the diagnostic value of tongue petechia in gynecology department tumor and dysmenorrheal, and provide theoretic basis for diagnosing gynecologic diseases by TCM. Methods A total of 169 patients with gynecologic tumor and dysmenorrheal were set as a treatment group, and a total of 211 patients who excluded the tongue signs of gynecologic tumor and dysmenorrheal were set as a control group. Tongue petechias were identified and compared between the two groups. Results ①Probability of tongue petechia showed a significantly higher value in the treatment group than the control group. (P<0.01) ; ②Probability of tongue petechia showed a significantly higher value of patients with tumor in the treatment group than patients with other gynecologic diseases in the control group (P< 0.05) ; ③Probability of tongue petechia showed a significantly higher value of patients with dysmenorrhea in the treatment group than patients with other gynecologic diseases in the control group (P<0.01). Conclusion ①Relevant researche of gynecologic tumor, dysmenorrheal and tongue petechia at the tip of tongue was based on TCM theory, diagnostics and clinical experiences. The data of this study has proven this diagnostic method.②Compared with other kinds of diseases, gynecologic tumor and dysmenorrheal showed a higher probability in the appearance of tongue petechia. Therefore, petechia at the tip of tongue can be used as one of diagnostic references for gynecologic tumor and dysmenorrheal.

Objective:To analyze the clinical and gene mutation characteristics of congenital disorder of glycosylation (CDG) caused by compound heterozygous mutation of the COG6 gene ( COG6-CDG). Methods:This study retrospectively analyzed the clinical data and genetic test results of a patient with COG6-CDG in Bayi Children's Hospital, the Seventh Affiliated Medical Center of Chinese PLA General Hospital, in August 2019. Literature was retrieved with keywords including COG6, COG6-CDG, congenital disorders of glycosylation typeⅡL and congenital disorders of glycosylationⅡL in China National Knowledge Infrastructure, Wanfang Database, VIP Database, PubMed, and Web of Science Database from the establishment to July 2020, to summarize the clinical and genetic characteristics of COG6-CDG. Results:(1) Case report: The 59-day-old baby boy, with a gestational age of 27 +5 weeks and birth weight of 1 180 g, presented with multi-system involvement on admission, including unidentified progressive hepatosplenomegaly with jaundice and ascites, persistent thrombocytopenia, microcephaly, hypotonia, hypohidrosis, hyperkeratosis, and recurrent hyperthermia, infection, and hypoglycemia, as well as dysfunctions of the heart, gastrointestinal tract, lungs, kidneys, ocular fundus, and the coagulation system. Despite given ventilator-assisted ventilation, anti-infection therapy, abdominal puncture and drainage, and blood transfusion, the patient still had an aggravated condition and eventually died of multiple organ failures 192 d after birth. Genetic analysis showed that the nuclear family carried compound heterozygous mutations in the COG6 gene (NM_020751.2), including missense mutations of c.662C>T(p.T221M) in exon 7 and c.443T>C(p.I148T) in exon 5, which were both novel mutations and originated from the mother and father, respectively. (2) Literature review: Eight related papers were retrieved, including 20 cases. The main manifestations were various degrees of nervous system abnormalities and growth retardation, complicated by abnormalities of the liver, heart, gastrointestinal tract, blood, immunity, teeth, and bones. All the reported cases suffered from mental and growth retardation, and nine deaths were reported. A total of 11 COG6 gene mutations were identified, and most of them were c.1167-24A>G splicing mutations in a deep intron (seven cases), followed by c.1646G>T (four cases) and c.511C>T (three cases). Conclusions:COG6-CDG commonly manifests as multi-system and multi-organ dysfunctions with poor prognosis. Gene detection is conducive to the accurate diagnosis of COG6-CDG. Our case carries compound heterozygous mutations of c.662C>T(p.T221M) and c.443T>C(p.I148T), which are unreported novel mutations.

Objective To evaluate the accuracy of dual-source CT (DSCT) in diagnosing the degree of coronary artery stenosis,and compare it with coronary artery angiography (CAG).Methods One hundred patients with suspected coronary heart disease,clinical diagnosis of angina pectoris,myocardial infarction and patients after percutaneous coronary intervention,were given DSCT and CAG examination.The CAG results were considered as the reference standard,and the results of the two examinations were compared.The sensitivity,specificity,positive predictive value and negative predictive value of DSCT in diagnosing coronary artery stenosis (mild ＜ 50％,middle 50％-75％,severe ≥75％) were computed.Results CAG displayed coronary artery 1320 segments,DSCT displayed coronary artery 1287 segments,the appreciable rate was 97.5％.The sensitivity of DSCT in diagnosing mild,middle and severe coronary artery stenosis were 64.4％ (105/163),88.7％ (235/265) and 97.4％ (187/192),specificity all were 99.7％ (617/619),positive predictive value were 98.1％ (105/107),99.2％ (235/237) and 98.9％ (187/189),negative predictive valne were 91.4％ (617/675),95.4％ (617/647) and 99.2％ (617/622).There was significant correlation between DSCT and CAG in diagnosing coronary artery stenosis by Spearman test (P＜ 0.01).Conclusions The results of DSCT have high accordance with CAG in diagnosing coronary artery stenosis.It is a kind of non-invasive examination with low cost,and could reduce the pain of invasive examination.So DSCT is a good choice in diagnosing coronary artery disease.

Third-class medical metrology station is being established in hospital for implementing national and military medical metrology law and regulations. The goal, significance and main functions are elaborated. It is suggested that the leader should take the initiative to grasp not only the overall design and the supplementary equipment construction, but the civil engineering technological design and the layout of metrology standard equipment. The general steps of establishing third-class medical metrology station are discussed.

Objectives:To study the effects of gene therapy with tissue type plasminogen activator(t PA)cDNA on the formation of thrombo embolism in vascular anastomotic sites. Methods:①The cDNA encoding t PA was amplified by RT PCR using the isolated total RNA as the template from the Bowes melanoma cells.②Recombinant plasmid pAdCMV t PA was cotransfected into 293 cells with pJMa 17 ,and the infectious but replication deficient AdCMV t PA was generated.③The rats were randomly divided into the control and treatment groups.11 0 nylone medical suture was applied to perform rat carotid artery end to end anastomoses.In the treatment group,AdCMV t PA solution was injected into the vascular anastomotic site while AdCMV (no containing t PA DNA) solution was injected into the control group. By means of RT PCR and chromogenic plasmin substrates,the following results were obtained. Results:①The t PA cDNA was successfully cloned and its eukaryotic expressing vector was constructed.②When the isolated RNA was performed with RT PCR,1.69 kb band appeared in the treatment group while the band could not be found in the control group.The t PA activity could be detected postoperatively on the 1st,2 nd,3 rd,4 th,5 th,6 th,7 th,10 th and 13 th day of the treatment,but could not be detected in the control group. Conclusions:The t PA gene can produce t PA having biological activity at anastomotic sites, possibly prevent the formation of thrombus embolism effectively and develop the anastomotic patency.