Objective To investigate the protective effects of the 14-3-3 protein overexpression on the injury of PC12 cell induced by MPP~+ and its mechanisms.Methods For expression in mammalial cells, pcDNA3.1(+)-14-3-3 plasmid was constructed and transfeeted into PC12 cell with Lipofectamine~(TM)2000. The overexpression of transfected 14-3-3 gene in PC12 cell was determined by immunofluorescence and Western blotting.The effects of 14-3-3 overexpressing on the cells viability,apoptotie ratio and the activity of superoxide dismutase(SOD)as well as glutathione peroxidase(GSH-Px)of PC 12 cell treated with MPP~+ were measured by MTT assay,flow cytometry analysis and microplate reader respectively.Results The expression of 14-3-3 protein in transfection group(1.19?0.06)increased evidently compared with control group(0.75?0.05).And the antioxidant enzyme activity assession,MTT assay and flow cytometry analysis shows that the overexpression of 14-3-3 protein elevates the activity of SOD(transfection group:(9.13? 0.41)U/mg protein,MPP~+ group:(6.45?0.52)U/mg protein)and GSH-Px(transfection group: (89.66?3.42)?mol/mg,protein MPP~+ group:(82.73?4.15)?mol/mg protein),increases the cell viability(transfection group:0.78?0.06,MPP~+ group:0.54?0.07),and inhibits cell apoptosis (transfeetion group:11.87%?3.26%,MPP~+ group:36.30%?2.39%)of PC12 induced by MPP~. Conclusion The overexpression of 14-3-3 protein could elevate the activity of antioxidant enzymes SOD and GSH-Px,reduce oxidant stress,alleviate MPP~+ toxicity,and thus inhibit the apoptosis of PC12 cell induced by MPP~+.

Objective To explore the value of 64-slice spiral CT angiography in diagnosis of pulmonary arteriovenous fistulas(PAVF).Methods 12 patients with suspect PAVF were performed by enhanced scan with 64-SliceCT.Two-dimensional and three-dimensional reformation were performed in all cases including coronal multiplanar reconstruction(MPR),maximum intensity projection(MIP),volume rendering(VR).Resnlts 12 cases were diagnosed as PAVF by 64-SliceCT angiography.Single PAVF in 7 cases,Multi-PAVF in 5cases.We found 20 lesions in 12 patients,Except direct multi-pulmonary arteriovenous fistulas in the area of near the heart in one case.Conclusion 64-slice spiral CT angiography is a useful and noninvasive means to identify PAVF and is also useful for therapeutic planning and postoperative follow-up.

Objective To investigate the correlation between ischemic stroke and both the risk factors and the degree of intracranial arterial stenosis, and provide evidence of preventing ischemic stroke. Methods Ninety patients with ischemic stroke were assessed by digital substraction angiography (DSA) and, accordingly, divided into group A (stenosis<30%) and group B (stenosis ≥30% or occlusion). The data about such risk factors as age, gender and family history, the levels of T-cholesterol (CHO), triacylglycerol (TG), high/low density lipoprotein cholesterin (H/LDL-C), apolipoprotein A1 (ApoA1), apolipoprotein B (ApoB) and lipoprotein A (Lpa) were recorded and analyzed; such diseases as hypertension, hyperlipemia, diabetes mellitus and coronary disease were taken into consideration. Results The incidence rate of intracranial arterial stenosis in these patients with ischemic stroke was 67.78%. Stenosis occurred most frequently in the middle cerebral arteries, less frequently in the intracranial segments of the internal carotid artery and the vertebral-basilar artery with the lowest occurrence in the posterior cerebral artery. Patients with ischemic cerebrovascular disease accompanied by high blood pressure, diabetes were more likely subjected to intracranial arterial stenosis; the regression coefficient, OR values, P values in patients with hypertension and diabetes were (1.659, 5.256 and 0.002) and (1.657, 5.241, and 0.046), respectively. The level of HDL-C (mmol/L) in the group B (0.99±0.30) was significantly lower than that in the group B (1.30±0.50, t=-3.603, P=0.001). Age, gender, smoking, stroke history, family history of cerebrovascular disease, the level of TC, TG, LDL-C, ApoA, ApoB, serum Lpa between the 2 groups showed no significant differences (P>0.05). Conclusion The major risk factors of intracranial arterial stenosis include hypertension, diabetes mellitus with HDL-C as its protective factors.

Objective To investigate the effects of 14-3-3 protein overexpression on the 1-methyl-4-phenylpyridinium (MPP(+)) induced pheochromocytoma (PC12) cell death and the potential mechanisms. Methods pcDNA3.1(+)-14-3-3 plasmids, which could be expressed in mammalian cell, were constructed and transfected into PC12 cells with Lipofectamine 2000. The expression of 14-3-3 protein, Bcl-2 protein, and BAD protein were determined by western blot. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, microplate reader, and flow cytometric analysis were used to measure cell viability, the caspase activity, and apoptotic ratio respectively. Results (1) The expression of 14-3-3 protein increased significantly three weeks after pcDNA3.1 (+)-14-3-3 plasmids transfected into PC12 cells. (2) MPP(+) caused a decrease of cell viability in a dose-dependent manner. At 100 mu mol/L MPP(+), cell viability reduced approximately 50%. (3) The caspase activity increased along with the MPP(+) concentrations rising and reached its maximum value (0.34 mu mol/mg protein) at 100 mu mol/L MPP(+). However caspase activity decreased significantly when the MPP(+) concentration exceeded 100 mu mol/L. (4) Overexpression of 14-3-3 protein decreased the apoptosis ratio of PC12 cells treated with 100 mu mol/L MPP(+) from 26.5% to 8.6%. (5) Bcl-2 protein tended to decrease but BAD protein tended to increase after treatment of PC12 cells with 100 mu mol/L MPP(+). Overexpression of 14-3-3 protein significantly increased the cellular level of Bcl-2 protein and decreased that of BAD protein. Conclusion Overexpression of 14-3-3 protein may reduce MPP(+)-induced apoptotic cell death in PC12 cells by up-regulating the Bcl-2 expression and down-regulating the BAD expression. These results may provide a promising target for treatment of Parkinson' s disease.

BACKGROUNDMany studies have shown that cancer cell differentiation and microvascular invasion play a principle role in cancer progression and metastasis, and non-invasive imaging techniques such as CT, MRI and US assessing the differentiation and the surgical resectibility and the prognosis of cancers are now of great importance. This study aimed to explore the correlation of triple-phase multi-slice CT scan with the histological differentiation and intratumor microvascular/lymphatic invasion of progressive gastric cancer.

METHODSThe present study included 64 patients with gastric cancer, all of whom underwent routinal and dual-phase contrast enhancement multi-slice CT examinations of the upper abdomen before surgery. The post-operative specimens were used for determination of histological differentiation, cancer cell invasion of intratumoral microvascular/lymphatic vessel identified by CD34 and D2-40 expression. Correlations between contrast enhancement ratio (CER) of triple-phase multi-slice CT scan in gastric cancer and histological differentiation as well as intratumoral microvascular/lymphatic invasion were compared and analyzed.

RESULTSThere was a significant correlation between CER of triple-phase CT scan in gastric cancer and tumor histological differentiation (P < 0.05). CER of the arterial phase in gastric cancer with intratumoral microvascular invasion was significantly higher than that without invasion (0.61 ± 0.28 vs. 0.46 ± 0.14, P < 0.05); CER of the arterial-parenchymal phase was significantly lower in gastric cancer with intratumoral microvascular invasion than that without invasion (1.81 ± 0.39 vs. 2.28 ± 0.80, P < 0.05). However, CER of the parenchymal phase in gastric cancer with intratumoral lymphatic invasion was significantly higher than that without invasion (1.25 ± 0.57 vs. 1.00 ± 0.35, P < 0.05).

CONCLUSIONSCER of triple-phase multi-slice CT scan in gastric cancer is closely correlated with intratumoral microvascular and lymphatic invasion, and also could be used as a marker for histological differentiation.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Immunohistochemistry ; In Vitro Techniques ; Lymphatic Metastasis ; diagnostic imaging ; Male ; Middle Aged ; Stomach Neoplasms ; diagnostic imaging ; metabolism ; pathology ; surgery ; Tomography, X-Ray Computed ; methods

OBJECTIVETo investigate the effect of therapeutic ultrasound-induced microbubble destruction on the microcirculation of rat skeletal muscle.

METHODSThirty SD rats were randomized into 5 groups (n=6), namely normal saline, microbubble, ultrasound, high-energy ultrasound microbubble and low-energy ultrasound microbubble groups. Before and after the treatments, the diameter and blood flow velocity in the microvessels in the skeletal muscle were measured, and the structural changes of the injured microvessels observed by electron microscopy.

RESULTSMicrobubble cavitation did not produce significant effect on the mean arterial pressure and diameter of microvessels in rat skeletal muscle (P>0.05), but the blood flow velocity was obviously lowered and blood flow volume reduced in the microvessels. The reduction of the flow velocity and blood flow volume and their subsequent recovery were associated with ultrasound energy, and in the low ultrasound energy group, the flow velocity and blood flow volume in the of venules recovered obviously after about 15 min, which, however, took approximately 1 h for the arterioles. In contrast, recovery of the flow velocity and blood flow volume in the microvessels took more than 2 h in the high ultrasound energy group. Cavitation resulted in endothelium cell rupture, widening of the endothelial interspace and entry of the red blood cells into the extravascular tissues as revealed by electron microscopy, but no rupture of the lining endothelium was observed 2 h after the treatment.

CONCLUSIONSEndothelium cell rupture induced by microbubble cavitation may affect the local microcirculation, and lower ultrasound energy exposure is associated with milder endothelial injury and more rapid recovery.

Animals ; Blood Flow Velocity ; Blood Vessels ; pathology ; physiopathology ; Endothelial Cells ; pathology ; ultrastructure ; Female ; Male ; Microbubbles ; Microcirculation ; Microscopy, Electron ; Microspheres ; Muscle, Skeletal ; blood supply ; Rats ; Rats, Sprague-Dawley ; Ultrasonics

OBJECTIVETo evaluate the role of enhanced recovery after surgery in treating cervical spondylotic myelopathy.

METHODSThe clinical data of 55 patients with cervical spondylotic myelopathy underwent surgical treatment from January 2012 to December 2015 were retrospectively analyzed. There were 30 males and 25 females, age from 36 to 71 years old with an average of (45.2±3.2) years, course of disease was for 1 to 12 months with an average of (4.5±1.8) months. The concept of enhanced recovery after surgery and perioperative management were applied to surgical treatment in 35 patients (study group), and the same period, 20 patients without strategy of enhanced recovery after surgery (control group). Thirty-eight patients were treated by anterior cervical discectomy decompression and fixation(ACDF), 17 patients were treated by posterior single-open door laminoplasty decompression. The activity time out of bed, hospitalization days after surgery were compared between two groups. Japanese Orthopaedic Association (JOA) score and visual analogue score(VAS) before operation, after operation at 1, 7, 30 days and 6, 12 months was respectively used to evaluate the neurological function and pain.

RESULTSAll the patients were followed up for 12 to 18 months with an average of (14.3±1.5) months. There was no significant difference in age, gender, surgical methods, preoperative VAS, JOA score between two groups (>0.05). The activity time out of bed was 3 to 8 h with an average of (5.54±1.54) h, postoperative hospitalization time was 3 to12 d with an average of (5.62±1.59) d in study group, while in control group, the activity time out of bed was 24 to 48 h with an average of (18.80±4.78) h, and postoperative hospitalization time was 7 to 17 d with an average of (9.85±1.94) d; there was significant difference between two groups (<0.01). There was significant difference in VAS and JOA scores between two groups at 1, 7, 30 d after operation (<0.01), and there was no significant difference at 6, 12 months after operation(>0.05). There were no neurologic function deterioration, hematoma, wound infection, internal fixation loosening and other complications in study group during hospitalization and following-up;there were 2 cases of superficial wound infection in the control group, who healed by dressing change for 2 weeks;there was no significant difference between two groups(>0.05).

CONCLUSIONSThe strategy of enhanced recovery after surgery in treating cervical spondylotic myelopathy can promote the early recovery, shorten the length of stay and improve the patient's degree of satisfaction.

The dynamic monitoring data of traditional Chinese medicine resources is one of the important tasks of the dynamic monitoring system of Chinese medicine resources,the system has formed a periodic monitoring data reporting mechanism. Data authenticity and accuracy are the basis for the sustainable and healthy development of Chinese medicine resources dynamic monitoring,information technology is an effective means to improve the efficiency of data reporting, reporting quality. Data production based on dynamic monitoring is of great significance for grasp the trend of change and development of Chinese medicine resources. In order to achieve the real-time control of changes to the national Chinese medicine resources, we build the Chinese medicine resources dynamic monitoring system. In order to solve the problems in practice, we have upgraded the fill system by using the data of GIS. In order to achieve the multidimensional, improve safety, practicality and standardization of the data, which laid a foundation for subsequent processing of data. The system can collect the information of the cultivation of Chinese herbal medicines,production and sales of daily reporting data, provide the Chinese herbal medicine market,fast growing industry environment such as positioning center. In this paper, the design and implementation of the system are expounded.According to the business requirements, we designed 12 forms, 98 collection indicators to meet the needs of dynamic monitoring of traditional Chinese medicine resources. This paper will introduce the development content, design and implementation, main function characteristics and application effect of the national Chinese medicine resources fill System. To explain the role that GIS technology plays in the system and how to realize the cultivation of Chinese herbal medicines, production and sales of daily reporting data, provide the Chinese herbal medicine market,fast growing industry environment such as positioning center,and information collecting.

A rich diversity of wild medicinal plant resources is distributed in China, but the breeding of new plant varieties of Chinese medicinal plants started late and the breeding level is relatively weak. Chinese medicinal plant resources are the foundation for new varieties breeding, and the plant variety rights(PVP) are of great significance for the protection and development of germplasm resources. However, most Chinese medicinal plants do not have a distinctness, uniformity, and stability(DUS) testing guideline. The Ministry of Agriculture and Rural Affairs has put 191 plant species(genera) on protection lists, of which only 30 are medicinal species(genera). At the same time, only 29 of 293 species(genera) plants in the Protection List of New Plant Varieties of the People's Republic of China(Forest and Grass) belong to Chinese medicinal plants. The number of PVP applications and authorization of Chinese medicinal plants is rare, and the composition of variety is unreasonable. Up to now, 29 species(genera) of DUS test guidelines for Chinese medicinal plants have been developed. Some basic problems in the breeding of new varieties of Chinese medicinal plants have appeared, such as the small number of new varieties and insufficient utilization of Chinese medicinal plant resources. This paper reviewed the current situation of breeding of new varieties of Chinese medicinal plants and the research progress of DUS test guidelines in China and discussed the application of biotechnology in the field of Chinese medicinal plant breeding and the existing problems in DUS testing. This paper guides the further application of DUS to protect and utilize the germplasm resources of Chinese medicinal plants.
Agriculture ; Biotechnology ; Plant Breeding ; Plants, Medicinal/genetics*

Chinese medicinal resources are the cornerstone of the sustainable development of traditional Chinese medicine industry. However, due to the fecundity of species, over-exploitation, and limitations of artificial cultivation, some medicinal plants are depleted and even endangered. Tissue culture, a breakthrough technology in the breeding of traditional Chinese medicinal materials, is not limited by time and space, and can allow the production on an annual basis, which plays an important role in the protection of Chinese medicinal resources. The present study reviewed the applications of tissue culture of medicinal plants in the field of Chinese medicinal resources, including rapid propagation of medicinal plant seedlings, breeding of novel high-yield and high-quality cultivars, construction of a genetic transformation system, and production of secondary metabolites. Meanwhile, the current challenges and suggestions for the future development of this field were also proposed.
Sustainable Development ; Plants, Medicinal/genetics* ; Plant Breeding ; Medicine, Chinese Traditional ; Technology