Objective To establish an optimum DNA extraction method for Chinese traditional medical herbs in order to meet necessary for DNA barcoding research.Methods Four Chinese traditional herbs, Glycyrrhiza uralensis Fisch, Phellodendron Chinensis Cortex, Cistanche tubulosa Wight and Cistanche deserticola Ma were chosen as the experimental materials, the DNA was extracted by 6 different kinds of DNA extraction method, including the improved method of high-salt combined low-pH,the improved method of SDS,CTAB method,PVP method,PlantZol Kit and Ezup Kit, the quality of DNA was investigated by ultraviolet spectrophotometry,agarose gel electrophoresis and PCR amplification by using specific primers of ITS2 and psbA-trnH. ResuIts The quality of the DNA was better than other four kinds of methods by the improved method of high-salt combined low-pH and Ezup Kit, the value of OD260/OD280 was between 1.7 ~1.9,the yield of DNA was the highest by the PlantZol kit , followed by the improved method of high-salt combined low-pH( P <0.05 ) , but the purity of DNA was poor by the PlantZol kit.The DNA electrophoresis tests showed that the DNA integrity of Glycyrrhiza uralensis Fisch and Cistanche tubulosa Wight were better with the improved method of high-salt combined low-pH, the improved SDS method, the CTAB method and the PlantZol kit.The DNA of Phellodendron Chinensis Cortex and Cistanche deserticola Ma were extracted by the six methods appeared diffuse status in the lanes.But only the improved method of high-salt combined low-pH could make the PCR amplification of the success rate 100% by using specific primers of ITS2 and trnH-psbA.ConcIusion The DNA extraction method of high-salt combined low-pH can be used to establish the Chinese DNA barcoding which has the advantages of lower cost, simpler procedure and less time.

Objective To construct in the extracellular matrix metalloproteinase inducer (EMMPRIN)glycosylation single point mutation plasmid,and to explore its relationship with tumor cell proliferation.Methods PCR point mutantion technology was used to construct the mutantion plasimid of EMMPRIN glycosylation single point.After successful mutation, the function of mutantion plasmids were detected. Western blotting was used to detect the expression of EMMPRIN protein,immunofluorescence method was used to detemine the morphological changes of the cells,and MTT assay was performed to detect the relationship between mutantion pasmid and tumor cell proliferation.Results Confirmed by restriction enzyme digestion and sequencing,the 44th,the 152th,and the 186th Asn were successfully mutated to Gln in the sequence of EMMPRIN;EMMPRIN/GFP (N44Q), EMMPRIN/GFP(N152Q),and EMMPRIN/ GFP (N186Q)glycosylation single point mutation plasmids were constructed.Compared with wild-type,thel morphology of the cells was significantly changed,the core division of mutant-type cells was significantly reduced, the number of filopodia was reduced. The results of MTT assay showed that the survival rate of the cells in wild-type group were significantly increased compared with control group (P<0.05 );the survival rates of the cells in EMMPRIN (N44Q) group, EMMPRIN (N152Q) group and EMMPRIN(N186Q)were significantly decreased compared with wild-type group(P<0.05).Conclusion Mutant-type EMMPRIN can inhibit the proliferation of tumor cells;with the duration increasing, the inhibitory effect is weakened.There is a correlation between EMMPRIN glycosylation and proliferation of tumor cells.

Objective To evaluate the clinical effect of postural reduction combined with miniincision screw-rod system and percutaneous kyphoplasty (PKP) in treating osteoporotic thoracolumbar burst fractures.Methods A retrospective case series study was performed for data of 35 patients with osteoporotic thoracolumbar burst fractures without neurological deficits undergone mini-incision screw-rod system fixation and PKP between January 2012 and January 2014.There were 14 males and 21 females,with a mean age of 63.2 years (range,50-72 years).Operation time,intraoperative blood loss,complications,visual analogue score (VAS),height of fractured vertebrae and kyphosis Cobb angle were recorded.Results Operation time was (49.6 ± 6.8) min,and intraoperative blood loss was (45.6 ±7.8)ml.All patients were followed up for 9-18 months (mean,13.5 months).No intraoperative or postoperative serious complications occurred,including intracanal cement leakage,breakage or loosening of the screws.VAS of back pain was decreased from (8.4 ± 1.1)points preoperatively to (3.5 ± 0.6)points postoperatively (P ＜ 0.05).Height of the fractured vertebrae was improved from (49.62％ ± 5.68)％ preoperatively to (86.64 ± 6.63) ％ postoperatively (P ＜ 0.05).Kyphosis Cobb angle was improved from (28.12 ± 1.06) °preoperatively to (5.15 ± 1.08) °postoperatively (P ＜0.05).At the final follow-up,VAS was further decrease and vertebral height and Cobb’ s showed a slight loss of correction.Conclusion Postural reduction combined with mini-incision screw-rod system and PKP can relieve back pain,restore the height of injured vertebrae,correct kyphotic deformity and reduce operation time and blood loss,indicating a minimally invasive,safe and effective procedure for treatment of osteoporotic thoracolumbar burst fractures.

Objective To investigate the feasibility and effectiveness of fast track surgery (FTS) in neurosurgery. Methods One hundred fifteen patients who underwent neurosurgery surgery in Henan Province People's Hospital from June 2012 to March 2014 were enrolled in this study. All the patients were divided into FTS group (62 cases) and the tra?ditional operation group (53 cases). The clinical index, postoperative hospital stay and hospitalization cost were compared between the two groups. Results The clinical index were significantly lower in FTS group than in traditional operation group (P<0.05). Length of hospital stay (days) and hospitalization cost of FTS group were significantly shorter and lower in FTS group compared with traditonal operation group (8±1 vs. 11±2 days and RMB 4.58 ±0.75 vs. 5.78 ±0.64 ten thou?sand, respectively) (P<0.05). Conclusion FTS in neurosurgery operation is an all-new concept for surgery which can ef?fectively reduce postoperative complications, shorten length of hospital stay, decrease hospitalization cost and promote postoperative recovery.

Objective To investigate the safety of the occipital condylar screw with vertical position and evaluate the selection strategy of the posterior approach of the posterior occipital condylar screw in Chinese people.Methods The clinical imaging data of 60 outpatients from September 2013 to September 2015 were retrospectively analyzed,36 male and 24 female,the average age was 41.6±9.2 (range from 25-58),Excluded occipitocervical injury,tumor and deformity patients.We built a three-dimensional digital model and simulated placing screw by utilizing CT data on Mimics software,after that we took the occipital condyle posterior medial and lateral midpoint as the entry point,then made 2 points equidistantly to the midpoint in vertical direction.We put 3.5 mm diameter virtual screws in 4 different conditions:largest cranial angle,smallest cranial angle,longest screw path and shortest screw path.Then we assessed the anatomical relationship between the screw and the hypoglossal canal or the atlanto-occipital joint by a three-dimensional window and measured the cranial angle,medial angle and length of screw path,then calculated the safety angle of the cranial angle,the successful rate of setting screw,and compared the safety of different screw points by 3-Matic software.Results 120 occipital condyles were obtained from the CT data of 60 patients by Mimics software.There was no significant difference in the data of the cranial angle,medial angle,safety range and length between both left and right sides.The obtained safe cranial angle of each point respectively was 20.9°±6.0° (lowest point),17.0°±6.2° (middle point),and 11.6°±7.1°(top point),obviously the largest angle was in the lower point and the smallest was in the top point.The difference was statistically significant.We then acquired the successful rates of different cranial angle of each point,the highest successful rate was 99.17％,96.67％,74.17％ in lowest,middle and top point when cranial angle were 3°or 4°,3°and 0°respectively.The successful rates of lower point and niddle point were significantly higher than the top point,and the difference was statistically significant.The medial angle parameters obtained were 34.41°±2.59°on left and 34.06°±2.44°on right,and there was no significant difference.The length parameters of the longest screw path acquired were 23.09± 1.47 mm,22.84± 1.40 mm and 23.15± 1.45 mm at top,middle and lowest entry point.The average value of shortest screw path of each point was 21 mm,and there was no significant difference among every entry point.Conclusion Among the occipital condyle posterior screw entering points,selecting the lower point can improve the success rate and safety;the change of nail enter point in the vertical direction has little effect on the length of the nail.We can increase the safety and reduce the risk of occipital condylar screw placement as far as possible through the three-dimensional digital technology.

Humans ; Child ; Child, Preschool ; Pinealoma ; Prognosis ; Brain Neoplasms/diagnosis* ; Pineal Gland

Objective To explore the biomechanical properties of posterior occipital condyle screws compared with common occipitocervical fusion internal fixation and it's impacts upon stress of hypoglossal canals.Methods Finite element models based on the occipitocervical CT data of one 28-year-old male healthy volunteer were built,including normal model,instability model,internal fixation model by occipital condyle screws,internal fixation model by occipital plate screws,and internal fixation model by transarticular screws.Fifty N gravity and 1.5 N · m torque were exerted upon the surface of occipital bone so that the models could perform lateral bending,flexion,extension,and rotational motions.The motion range and stress distribution of internal fixation were compared under varying conditions among different occipitocervical fusion models.In addition,the impact of occipital condyle screw upon hypoglossal canals was examined.Results Compared with instability model,the motion range in the internal fixation model by occipital condyle screws declined by 96.8％,95.6％,95.0％ and 98.5％ respectively in lateral bending,flexion,extension and rotation.In the internal fixation by occipital plate screws,the motion range decreased by 96.3％,95.7％,98.4％ and 99.6％ respectively.In the internal fixation by transarticular screws,the motion range exhibited a decline of 95.7％,94.0％,94.3％ and 98.9％,respectively.The stress peaks in the occipital condyle screw were 192.4 MPa,201.6 MPa and 187.6 MPa under lateral bending,flexion,and rotation conditions,respectively.The stress peaks in the occipital plate screw were 279.6 MPa,213.7 MPa,and 154.1 MPa,respectively.The stress peaks in the transarticular screw were 232.4 MPa,220.9 MPa,and 224.5 MPa,respectively.The stress impact peak of occipital condyle screw on the hypoglossal canals wall was 12.96 MPa,and the content deformationunder the hypoglossal canal was 0.64％.Conclusions The occipital condyle screw internal fixation has similar stability with common occipitocervical fusion fixations.The occipital condyle screw has more uniform stress distribution and less effect on the hypoglossal canals,and hence is safe and reliable as anchor point on the cranial side in occipitocervical fusion.

Objective To assess the effects of axial spinous process-muscle-vascellum complex transplantation for posterior atlantoarial fusion.Methods Data of 27 cases with altantoarial disease who were treated by posterior atlantoarial fusion using axial spinous process-muscle-vascellum complex transplantation from June 2015 to June 2016 were retrospectively analyzed.There were 19 males and 8 females aged from 9 to 68 years old (mean,41.0±15.4 years old).Two cases were diagnosed with atlanto-axial instability.Fourteen cases were diagnosed with atlas fracture and eleven cases were diagnosed atlanto-axial fracture.All the 27 patients suffered from neck pain or limitations of cervical motion.All patients were assessed clinically by atlantoaxial reduction and bone graft fusion.The pre-operative and post-operative atlanto-dens interval (ADI),visual analogue scale (VAS),Japanese Orthopaedic Association scores (JOA),improvement rate of JOA score and axial symptoms were measured and statistically analyzed.Complications were recorded.Clinical outcome of latest follow-up was evaluated by X-ray and CT scan.Results The time of operation was 2.0-2.5 h and blood loss was 150-300 ml.All the patients were followed-up for 9 to 18 months (mean,11.5±2.1 months).The VAS of neck pain improved from 3.6±2.7 (range,2.0-5.0) pre-operatively to 1.4±0.2 (range,0-2.0) 12 months postoperatively (P=0.000).The JOA score improved from 11.7± 1.9 (range,10.0-15.0) pre-operatively to 15.3±0.6 (range,14.0-17.0)12 months post-operatively (P=0.000).The improvement rate of JOA score at the latest follow-up was 54.1％± 12.4％,including 23 cases (85.19％) excellent,and 4 cases (14.81％) good.The results of axial symptoms were no-symptom in 22 cases (81.48％) and mild symptoms in 5 cases (18.52％).Postoperative cervical spine X-ray and CT showed that the sagittal cervical spine alignment was restored.There was statistically significant difference between ADI of 4.3±1.1 mm (range,3.9-4.5 mm) pre-operatively to 2.5± 0.4 mm (range,2.1-2.6 mm) 12 months post-operatively,which was improved significantly (P=0.000).There were no complications found during the follow-up.Conclusion The application of axial spinous process-muscle-vascellum complex transplantation for posterior atlantoaxial fixation can preserve the dynamic function of muscles and reduce the postoperative pain,as well as avoid donor site morbidity.

Childhood asthma is one of the most-common chronic respiratory diseases in the world.The incidence of asthma is increasing, and therefore, controlling the occurrence and development of asthma has been the current research hotspot.A growing number of evidences have shown that the changes of maternal internal and uterine environment during pregnancy are related to the occurrence of asthma in offspring.The influence of maternal mental, disease, nutrition, environment, drug exposure on the occurrence of asthma in offspring were summarized, providing a new idea for understanding the mechanism, prevention and treatment of children asthma.

Objective:To discuss the regulatory effect of berberine(BBR)on fatty acids in the human glioma T98G cells and its effect on the cell proliferation,migration,and invasion,and to clarify its potential mechanism.Methods:The T98G cells at logarithmic growth phase were divided into control group and different concentrations(25,50,and 100 mg·L-1)of BBR groups.Cell wound healing assay was used to detect the migration rates of the cells in various groups;Transwell chamber assay was used to detect the invasion rates of the cells in various groups.The T98G cells at logarithmic growth phase were divided into control group and 100 mg·L-1 BBR group,and Mass spectrometry was used to detect the fatty acid contents in the cells in two groups.The T98G cells at logarithmic growth phase were divided into control group and different concentrations(50,100,and 150 mg·L-1)of BBR groups;Western blotting method was used to detect the expression levels of phosphatidylinositol 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(AKT),phosphorylated AKT(p-AKT),sterol regulatory element-binding protein 1(SREBP-1),and fatty acid synthase(FASN)in the cells in various groups.The expression of FASN was suppressed by gene silencing technology,and the T98G cells at logarithmic growth phase were divided into control group,shFASN1 group,and shFASN2 group.Western blotting method was used to detect the expression levels of FASN protein in the cells in various groups;clone formation assay was used to detect the clone formation of the cells in various groups;cell wound healing assay was used to detect the migration rates of the cells in various groups.Results:Compared with control group,the migration rates and invasion rates of the cells in different concentrations of BBR groups were decreased in a concentration-dependent manner(P<0.01),and the fatty acid content in the cells in 100 mg·L-1 BBR group was significantly decreased(P<0.01).Compared with control group,the expression levels of p-PI3K,p-AKT,SREBP-1,and FASN proteins in the cells in 150 mg·L-1 BBR group were significantly decreased(P<0.05 or P<0.01),and the expression level of SREBP-1 protein in the cells in 100 and 150 mg·L-1 BBR groups were significantly decreased(P<0.01).After suppression of FASN expression,compared with control group,the expression levels of FASN protein in the cells in shFASN1 and shFASN2 groups were significantly decreased(P<0.01),and the expression level of FASN protein in the cells in shFASN2 group was lower than that in shFASN1 group(P<0.05);compared with control group,the numbers of clone formation and migration rates of the cells in shFASN1 and shFASN2 groups were significantly decreased(P<0.01),and the migration rate of the cells in shFASN2 group was significantly lower than that in shFASN1 group(P<0.05).Conclusion:BBR interferes with fatty acid synthesis in the glioma T98G cells by reducing the expression of the PI3K/AKT/SREBP-1/FASN pathway related proteins,and decrease their migration and invasion capabilities.