Objective To investigate the effects of mild hypothermia ( MH ) on the expression of my- eloperoxidase(MPO) and cyclooxygenase 2 (COX 2) in rats after cerebral ischemia and reperfusion (CIR). Methods Forty-eight Wistar rats were randomly divided into four control groups (n=6 in each) and four MH groups (n=6 in each).CIR models were established by suture occlusion of the left middle cerebral artery.The rats in the MH groups,but not in the control groups,were treated with MH.Rats were killed at 4 h,8 h,12 h and 16 h after CIR.MPO expression was measured,along with the expression of COX 2 as measured by Western blot- ting and immunohistochemical methods.Results Compared with the control groups,MPO activity and the COX 2 expression in the cortex and striatum were significantly lower in all the MH groups at 4 h,8 h,12 h and 16 h after CIR.Conclusion MH treatment can protect neurons by decreasing MPO activity and COX 2 expression,allevia- ting inflammation and reducing secondary injuries after CIR.

Objective To investigate the present status and developments of Chinese military neurosurgery, make an objective analysis about the achievements and deficiencies, and propose a developing trend for the field of Chinese military neurosurgery. Methods The scientific literatures of neurosurgery published domestically and abroad in recent 5 years were partially reviewed. The advances and achievements in basic researches and clinical works of neurosurgery acquired during the period of the 11th Five Year Plan were analyzed. Results During the period of the 11th Five Year Plan, Chinese military neurosurgery was still in the lead position in traditional areas such as traumatic brain injury. In the meantime, great achievements were obtained in the researches of brain and spinal cord tumors, endovascular treatment and functional neurosurgery, and 47 items of Army Science and Technology Advancement Prizes or Military Medical Achievement Awards were acquired. A great progress was made in personnel training. However, unbalanced development in neurosurgery field still existed within military departments, between military and civil departments and between different regions, so an enhanced technical interchange and cooperation is needed. Conclusions On the coming period, the research work of military neurosurgery should focuse on basic and clinical research of gene therapy to keep in step with neurosurgical trends in the world. On the other hand, attention should be paid to technological innovation with military features such as professional and general treatment of traumatic brain injury and researches on the development of protective equipments.

Objective To evaluate the role of combination of three-vessel views in ultrasound diagnosis of fetal aortic arch and pulmonary arterial branch anomalies.Methods The data of 66 cases of fetal aortic arch and pulmonary arterial branch anomalies were retrospectively analysed,including echocardiographic data,autopsy and operation records and postnatal follow-up results.Echocardiogaphic features and display frequencies on three vessels and trachea view(3VT),three vessels view(3VV)and three vessels and pulmonary arterial branches view(3VP)were summarized.Results There were 52 cases of aortic arch abnormalities,including 32 cases of right aortic arch with left ductus arteriosus,4 cases of double aortic arch,7 cases of aberrant right subclavian artery,6 cases of coarctation of aorta and 3 cases of interruption of aortic arch.Fourteen cases were diagnosed pulmonary aterial branch abnormalities,including 1 0 cases of crossed pulmonary arteries,2 cases of anomalous origin of left pulmonary artery from aortic arch and 2 cases of pulmonary artery sling.The display frequencies of fetal aortic arch and pulmonary arterial branch anomalies on 3VT,3VV and 3VP were 80.3%,19.7% and 39.4%.Conclusions The combination of three-vessel views is of great value in prenatal diagnosis of fetal aortic arch and pulmonary arterial branch anomalies.

Objective Yangqing Chenfei Formula - To investigate the effect of (YCF) on epithelial mesenchymal transition (EMT) Methods in lung tissues of silicosis model rats. Specific pathogen free adult male SD rats were randomly divided into control group, model group, tetrandrine group and YCF group, with eight rats in each group. The rats in the model group, tetrandrine group and YCF group were intratracheally injected with 1.00 mL of silica suspension with a mass concentration of 50.0 g/L, and the rats in the control group were given an equal volume of 0.9% sodium chloride solution. On the 15th day after modeling, the tetrandrine group was given tetrandrine at a dose of 27.0 mg/kg body weight, the YCF group was given YCF with a dose of 8.91 g/kg body weight, while both the control group and model group were given 2.00 mL 0.9% sodium chloride solution. Gavage wasperformed twice a day in the morning and evening for 14 days. On day 29 of the experiment, after evaluating the tidal volume, - functional residual volume (FRC) and vital capacity of rats in each group, lung tissues were collected, and hematoxylin eosin staining and Masson staining were performed to examine the histopathological changes, and the fibrosis score was evaluated. - - Hydroxyproline level was detected by colorimetry. The expression of type Ⅰ collagen (COL Ⅰ), type Ⅲ collagen (COL Ⅲ), - - - - - - E cadherin (E Cad), N cadherin (N Cad) and α smooth muscle actin (α SMA) protein was detected by immunohistochemistry. - The expression of epithelial cell adhesion molecule (EpCAM) and fibroblast specific protein 1 (FSP 1) was detected by Results immunofluorescence. The lung structure was intact and the alveolar structure was normal in the control group. The alveolar structure was destroyed, the alveolar wall was thickened, and cellular nodules were observed/n the model group. The lung tissue lesions of rats in the tetrandrine group and YCF group were reduced compared with that in the model group, and there was no difference in the degree of lesions between the two groups. The tidal volume, FRC and vital capacity of rats in model P< - P< group decreased (all 0.05), the relative expression of E Cad protein in lung tissue decreased ( 0.05), the fibrosis score and - - - - the level of hydroxyproline, the protein relative expression of COL Ⅰ, COL Ⅲ, N Cad and α SMA in lung tissue increased (all P< - 0.05), while the fluorescence intensity of EpCAM protein decreased, and that of FSP 1 protein increased compared with the P< control group. The tidal volume, FRC and vital capacity of rats in tetrandrine and YCF groups increased (all 0.05), the fibrosis - - - score and the level of hydroxyproline, the protein relative expression of COL Ⅰ, N Cad and α SMA in lung tissue decreased (all P< - P< 0.05), the relative expression of E Cad protein in lung tissues increased ( 0.05), while the EpCAM protein fluorescence - intensity increased and FSP 1 protein fluorescence intensity decreased compared with the model group. The relative expression - P< Conclusion of N Cad protein in lung tissues of YCF group was lower than that of the tetrandrine group ( 0.05). YCF can - improve the lung function, alleviate collagen deposition in lung tissues, and inhibit the epithelial mesenchymal transition in silicosis model rats, and then attenuates the progression of silicotic fibrosis.

Cancer is considered as one of the major diseases endangering human health in the world, it is urgent to find a safer and more efficient treatment for cancer therapy. Gene therapy with ribonucleic acid (RNA) drugs could regulate the expression of tumor related genes, and exhibit good anti-tumor therapeutic potential in preclinical and clinical trials. Based on the differences between tumor tissues and normal tissues in microenvironment signal characteristics such as pH, specific enzyme concentration or redox gradient, various microenvironment responsive nanocarriers had been studied and developed to deliver RNA drugs to tumor tissues and cells, improving the anti-tumor efficacy of RNA drugs and reducing toxic and side effects. This paper reviews the pathophysiological characteristics of tumor microenvironment and various strategies of tumor microenvironment responsive nanocarriers, in order to provide reference for the design of safe and efficient RNA drug delivery system for cancer therapy.

Objective To review the clinical characteristics and treatment of type 1 diabetes mellitus (T1DM) in children. Methods The clinical data of 103 children with T1DM admitted to our hospital from Februry 2002 to August 2010 were retrospectively analyzed. Thirty one cases with diabetic ketoacidosis (KDA) were treated with continuous insulin pump (group A) or basal-bolus insulin therapy (group B). The differences in blood glucose control time, the rate of hypoglycemic episodes, glucose fluctuation, fasting blood glucose (FBG), 2 h postprandial blood glucose (2 hPBG), insulin dosage, the time of urine acetone bodies disappear and length of stay were compared in two groups. Results The age of 103 children with T1DM was from 38 d to 15. 33 y with an average of (8 ±3) y; most of them was 7 - 10 y (47, 45.6% ). Seventy eight children were first diagnosed accounting for 75.7%; boys accounted for 55.3% of total. Fifty one cases (49.5%) were diagnosed in winter and spring and 67 (65.2%) had infections, most of them were respiratory tract and gastrointestinal infections. Sixty two cases (60. 2% )presented as diabetic ketoacidosis at the first onset, including 4 cases (3.9%) with cerebral edema. Some patients were complicated by Hashimoto's thyroiditis, hyperthyroidism, SLE and other autoimmune diseases.Among 31 cases with ketoacidosis the FBG and PBG were decreased significantly after treatment, there were no significant differences between two groups (P ＞ 0. 05 ). Compared to group B the correction time of DKA and urine acetone bodies was shorter, and reached the targeted glucose levels more quickly, the frequency of blood fluctuation and the hypoglycemia was significantly lower, the length of stay was shorter, and the daily dose of insulin was lower in group A; the differences between two groups were statistically significant ( P ＜0. 05 or P ＜0. 01 ). Conclusions The clinical symptoms at first onset of T1 DM in children are not typical,and likely to be combined with DKA; infection may be one of the inducing factors for DKA. Continuous subcutaneous insulin infusion with pump can control the blood glucose more effectively and equably, and are convenient for use by children; so it is a better treatment option for type 1 diabetes mellitus in children.

The objectives of this study were to investigate the effects of the CRISPR/Cas9 system mediated by the HPV pseudotype virus on SiHa cytobiology behavior by cutting the HPV16 E6 gene selectively and to explore the role of this system in the treatment of cervical cancer.After designing specific gRNA sequences targeting HPV 16 E6,generating hCas9-EGFP and E6-gRNA-RFP plasmids,and preparing the pseudovirus of HPV16 carrying E6-gRNA and Cas9 plasmids,we determined the titer of the pseudotype virus using the TCID50 method.We obtained the pseudotype virus of HPV16 carrying E6-gRNA and Cas9 plasmids to transfect cervical cancer SiHa cells.Experimental subjects were divided into control group,empty virus group,E6-gRNA transfected group,Cas9 transfected group and Cas9+E6-gRNA transfected group.The molecular size of the cutting sequence was detected using the T7E1 enzyme digestion method and agarose gel electrophoresis,and the cleavage function of CRISPR/Cas9 on the E6 gene was determined at the same time.RT-PCR and Western blotting were performed to detect the mRNA and protein expression levels of E6 in all the groups;the Transwell cell migration assay was performed to detect the cell migration ability and metastasis in all groups.Heterotopic transplantation tumors were incorporated into mice and were used to investigate the effects of the CRISPR/Cas9 system mediated by the HPV pseudovirus on the tumorigenic ability of SiHa cells by selectively cutting HPV16 E6.The HPV16 pseudotype virus carrying E6-gRNA and Cas9 plasmids could successfully infect SiHa cells,and there were two cutting zones in the Cas9+E6-gRNA transfected group.However,the empty virus group,E6-gRNA transfected group and Cas9 transfected group had no corresponding zone.Compared with those in the control group,the empty virus group,E6-gRNA transfected group and Cas9 transfected group,the mRNA and protein expression levels of E6 in SiHa cells were downregulated in the Cas9+E6-gRNA transfected group (P＜0.01).In addition,the proliferation and migration abilities of SiHa cells were significantly inhibited (P＜0.01).There were no significant differences among the other groups.In contrast to the control group,the HPV pseudotype virus carrying E6-gRNA and Cas9 plasmids could significantly delay the growth of tumor cells of the ectopic tumor transplantation model (P＜0.01).The CRISPR/Cas9 system mediated by the HPV pseudotype virus to knockout E6 gene expression exhibited a clear inhibitory effect on the biological function of SiHa cells,which indicated that knocking out the E6 gene using the CRISPR/Cas9 system mediated by the HPV pseudotype virus had a potential effect of eliminating HPV infection and inhibiting the growth of HPV-related tumors.Taken together,these findings provide insight into a new treatment strategy for the prevention and treatment of hr-HPV infected disease,particularly in HPV-related tumors.

Objective To evaluate the diagnostic value of 64 multidetector-row CT angiography for internal carotid artery(ICA)stenosis and the application in the follow-up of carotid endarterectomy and percutaneous transluminal stenting.Methods Forty transient ischemie attack(TIA)patients with interpretable CTA and DSA of the cervical carotid arteries were selected from May 2005 to December 2005. This yielded a total of 80 vessels.The CTA curved planar reformations(CPR)and DSA images referenced to the distal cervical internal carotid were graded by two senior neuroradiologists blindly,according to the North American Symptomatic Carotid Endarterectomy Trial(NASCET)guidelines.The paired-t test was used to verify the statistical significant difference between pre-operating and post-operating of carotid endarterectomy or percutaneous transluminal stenting in measuring the vascular diameter and area of cross section using CTA.Results When the 70% stenosis was used as the cut-off value,the seasitivity,specificity,negative predictive value,and the positive predicting value were 97%,95%,95%,and 98%,respectively.There was statistically significant difference in measuring the vascular diameter(P

Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
Aminoacyltransferases ; metabolism ; Bacterial Proteins ; metabolism ; Cyclization ; Cysteine Endopeptidases ; metabolism ; Enzymes, Immobilized ; metabolism ; Kinetics ; Peptides ; metabolism ; Peptides, Cyclic ; biosynthesis ; Staphylococcus aureus ; enzymology

Objective To investigate the clinical value of prenatal ultrasonography in diagnosing fetal aortic atresia.Methods Ultrasound images of 9 cases with fetal aortic atresia confirmed by autopsy were retrospectively analysed and summarized.Results Fetal echocardiography showed ascending aorta and aortic arch were thin in seven cases and unclear in two cases.Color Doppler flow imaging showed fetal aorta no blood connection with left ventricle and reverse blood flow in aortic arch among all 9 cases.There were 6 cases associated with hypoplastic left heart syndrome,two cases associated with ventricular septal defect and well-developed left ventricle,one case complicated with complete atrioventricular septal defect.Nine cases were detected at average gestational age (26.0 ± 2.6) weeks,minimum gestational age was only 16 weeks pregnant.Conclusions Prenatal ultrasound diagnosis of fetal aortic atresia has important clinical value.Ascending aorta and aortic arch are thin or unclear,no blood flow connection between fetal aorta and left ventricle and reverse blood flow in aortic arch are major diagnostic evidences.Color Doppler flow imaging is helpful for early diagnosis.