Objective To investigate the damage of hippocampal neurons induced by chronic cerebral ischemia in the rats,and to clarify its mechanism.Methods 90 healthy adult male SD rats were randomly divided into sham operation group (n=30 ) and experimental group (n=60 ). The chronic cerebral ischemia rat models were established by permanently ligating the common carotid arteries on both sides of the rats in experimental group.The rats in sham operation group were established by incising the cervical median,only the common carotid arteries on both sides were separated without ligating. The rats in sham operation group and experimental group were respectively sacrificed at the 7th,14th and 21st day after operation.At each time point 10 rats in sham operation group and 20 rats in experimental group were selected and sacrificed.Immunohistochemistry staining was used to observe the dynamic changes of the expression levels of choline acetyltransferase(ChAT)in hippocampus neurons, and TA-FE method was used to observe the dynamic changes of hippocampal microvascualr architecture. MiVnt image analytical system was used to quantitatively analyze the immunohistochemistry result,the microvessel density (MVD)and micorvessel area density (MVA)of horizontal part of hippocampus in the rats. Results Compared with sham operation group,the ChAT expression levels in hippocampus neurons of the rats in experimental group at different time points were significantly decreased(P<0.05);and with the prolongation of time,the ChAT expression levels were gradually decreased;the ChAT expression level in 14-day experimental group was significantly lower than that in 7-day experimental group (P<0.05 );the ChAT expression level in 21-day experimental group was significantly lower than those in 7-day and 14-day experimental groups(P<0.05).The MVD and MVA of hippocampus of the rats in experimental group at different time points were obviously decreased compared with sham operation group(P<0.05);the MVA was gradually decreased with the prolongation of time, and the MVA of hippocampus of the rats in 21-day and 14-day experimental groups were obviously decreased compared with 7-day experimental group(P<0.05);the MVD was gradually decreased with the prolongation of time,the MVD of hippocampus of the rats in 21-day and 14-day experimental groups was obviously decreased compared with 7-day experimental group(P<0.05);the MVD of hippocampus of the rats in 21-day experimental group was obviously decreased compared with 14-day experimental group(P<0.05).Conclusion Chronic cerebral ischemia can lead to the progressive decrease of the ChAT expression level,MVD and MVA of hippocampus of the rats to aggravate gradually the learning and memory dysfunction, which may be one of the reasons of vascular dementia.

Objective: To study the chemical constituents of Potentilla multifida L. Methods: Chemical constituents were isolated by the repeated silica gel chromatography and Sephadex LH 20, and their structures were identified by the spectral analysis. Results: Five compounds were obtained as follows: 3?,24 dihydroxyl urs 12 ene (1), ursolic acid (2), euscaphic acid (3), tormentic acid (4), and epihedaragenin (5). Conclusion: Five compounds were isolated from this plant for the first time. Compounds 1 and 5 were isolated from this genus for the first time. Compound 1 was a new natural product.

Objective To explore the clinical effect of neoadjuvant chemo-radiation combined with thoracoscopic and laparoscopic three-incision esophagectomy for esophageal cancer.Methods The retrospective cohort study was conducted.The clinicopathological data of 111 esophageal cancer patients who underwent neoadjuvant chemo-radiation combined with thoracoscopic and laparoscopic three-incision esophagectomy in the Cancer Hospital of Chinese Academy of Medical Sciences between January 2010 and December 2016 were collected.Among 111 patients,68 with interval time between neoadjuvant chemo-radiation and thoracoscopic and laparoscopic three-incision esophagectomy ＜ 8 weeks were allocated into the ＜ 8 weeks group and 43 with interval time between neoadjuvant chemo-radiation and thoracoscopic and laparoscopic three-incision esophagectomy ≥8 weeks were allocated into the ≥8 weeks group.Patients underwent preoperative radiotherapy and chemotherapy with TP regimen,and then underwent selective thoracoscopic and laparoscopic three-incision esophagectomy.Observation indicators:(1) neoadjuvant chemo-radiation situations;(2) surgical and postoperative situations;(3) follow-up.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival of patients and tumor recurrence or metastasis up to March 2017.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed using the t test.Count data were analyzed using the chi-square test or Fisher exact probability.Comparison of ranked data was done by the nonparametric test.Results (1) Neoadjuvant chemo-radiation situations:all the patients underwent neoadjuvant chemo-radiation,without severe adverse reaction.Number of patients with complete remission based on oncopathology were 34 in the ＜8 weeks group and 15 in the ≥ 8 weeks group,with no statistically significant difference between the 2 groups (x2=2.441,P＞0.05).(2) Surgical and postoperative situations:all the patients underwent successful thoracoscopic and laparoscopic three-incision esophagectomy,with negative surgical margins.Operation time,volume of intraoperative blood loss,number of lymph node dissected,time of postoperative intrathoracic drainagetube removal,time of postoperative neck drainage-tube removal,hoarseness,upper gastrointestinal hemorrhage,anastomotic fistula,respiratory complication,pleural effusion and empyema,cardiovascular complication,incision abnormal healing,death within postoperative 30 days and duration of hospital stay were (354±103)minutes,(400± 76)mL,19±4,(11±4)days,(4.9±1.6)days,5,1,12,3,6,5,8,0,(19± 17) days in the ＜ 8 weeks group and (343±92) minutes,(392±51)mL,19±3,(12±6)days,(4.5±1.0)days,2,0,7,5,3,4,3,2,(18± 11) days in the ≥ 8 weeks group,respectively,with no statistically significant difference between the 2 groups (t =1.080,0.569,0.326,1.223,1.286,x2=0.029,0.035,1.114,0.000,0.000,0.246,t=0.315,P＞ 0.05).(3) Follow-up:90 of 111 patients were followed up for 3-82 months,with a median time of 25 months,including 55 in the ＜8 weeks group and 35 in the ≥8 weeks group.During follow-up,death and tumor recurrence were detected in 9,11 patients in the ＜8 weeks group and 6,11 patients in the ≥ 8 weeks group,respectively.Conclusion Neoadjuvant chemo-radiation combined with thoracoscopic and laparoscopic three-incision esophagectomy is safe and effective,and it doesn't increase the perioperative risks based on preoperative 8-week interval time.

Objective To analyze the clinical and radiological features of retroperitoneal fibrosis (RPF), and to deepen the understanding of this unusual disease and improve the diagnostic level at the early stage. Methods Fourteen cases (10 males and 4 females, mean age 45.8 years) of pathologically diagnosed RPF from January 1990 to June 2004 were summarized. The clinical and radiological performance of the cases were analyzed. All patients received non-contrast CT scanning, 10 of them underwent enhanced CT scanning as well. 8 patients received MRI, 10 patients received IVP examination, and 11 received B-ultrasound. Results (1)The very first symptoms usually included back pain, bellyache (10 cases), or urinary tract obstruction (3 cases), with increase of ESR, IgG, CRP value and abnormal renal function.(2)The result of radiological examination showed that 11 lesions of the 14 cases located at retroperitoneum. Ten cases were mass type and 4 cases were diffuse type. Non-contrast CT scanning revealed soft tissue mass at retroperitoneum with inhomogenous or homogenous density. After contrast medium injection the lesions were enhanced with different extent. MRI results showed that the lesions presented low signal in T_1WI, while in T_2WI the signals had no obvious coherence but were different from one case to another. Conclusion Radiological examination is one of the important methods for diagnosis of RPF. Based on the different characteristics of RPF in CT and MRI, together with the clinical findings, we will get valuable references for staging and follow-up of RPF.

BACKGROUND:Shear stress can directly mediate the expression of endothelial cells, especially some cytokine genes whose codes are related to angiogenesis. Otherwise, flow shear stress of blood plays an importantly biological role in regulating vascular structure and function.OBJECTIVE: To observe the effects of laminar flow shear stress on the proliferation of vascular endothelial cells and the expression of protooncogene c-myc in human cerebral arteriovenous malformation.DESIGN: Randomized controlled study.SETTING: Department of Neurosurgery, Shenyang General Hospital of Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out in the Neuromedical Institute, General Hospital of Shenyang Military Area Command of Chinese PLA from November 2006 to February 2007. Fresh samples of human cerebral arteriovenous malformation were derived from 20 patients who were of grade Spetzler Ⅱ -Ⅲ and received resection of human cerebral arteriovenous malformation in the Department of Neurosurgery, General Hospital of Shenyang Military Area Command of Chinese PLA in 2006. All cases were diagnosed with whole-brain angiography before operation. The main reagents and equipments were detailed as follows: M199 culture media (Gilbco BRL), quality fetal bovine serum (HyClone), endothelial cell growth supplement (ECGS; Sigma, USA), CO2 incubator (Forma Scientific, USA), flow cytometry analysis of cell cycle kit (BD Company), flow cytometer (FACS Calibur, BD Company), rat-anti-human c-myc monoclonal antibody (Santa Cruz Company, USA), and reverse transcription polymerase chain reaction (RT-PCR) kit (Promega).METHODS: Tissue explants adherent method was used to culture vascular endothelial cells of human cerebral arteriovenous malformation, and then the cells were classified into 4 groups based on degree of shear stress, including control group, low shear stress group, moderate shear stress group and high shear stress group. Cultured endothelial cells of human cerebral arteriovenous malformation were put in a parallel plate flow chamber. In addition, cells in the low,moderate and high shear stress groups were stressed by low, moderate and high shear stress for 8 hours, respectively.However, shear stress in the control group was 0 Pa. Flow cytometry was used to measure proliferation index, and the expression of c-myc protein and c-myc mRNA were determined by immnocytochemistry and RT-PCR analysis respectively.MAIN OUTCOME MEASURES: Expressions of c-myc protein and c-myc mRNA and proliferation index in endothelial cells under various degrees of shear stress.RESULTS: ① Proliferation index: Proliferation index was higher in the moderate and high shear stress groups than that in the control group (P ＜ 0.05, 0.01). ② Expression of c-myc protein: Immuneposjtjve expression of c-myc protein was gradually increased with the increase of shear stress and there were significant differences in the three shear stress groups as compared with control group (P ＜ 0.05-0.01). ③ Expression of c-myc mRNA: Proliferation index of endothelial cells was higher in the low and moderate shear stress groups than that in the control group (P ＜ 0.05).CONCLUSION: Flow shear stress can induce expression of c-myc and activate expression of c-myc gene based on gene transcription so as to promote the proliferation of vascular endothelial cells in human cerebral arteriovenous malformation

BACKGROUND:Antisense gene therapy offers immense promise for the management of human cerebral arteriovenous malformation through inhibiting expression of vascular endothelial growth factor and angiogenesis in endothelial cells.OBJECTIVE: To observe the inhibitory effect of vascular endothelial growth factor-antisense oligonucleotide (VEGF-ASODN) on the proliferation of vascular endothelial cells in human cerebral arteriovenous malformation.DESIGN: Observational contrast study.SETTING: Department of Neurosurgery, General Hospital of Shenyang Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out in the Neuromedical Institute, General Hospital of Shenyang Military Area Command of Chinese PLA from August to December 2006. A total of 18 patients with human cerebral arteriovenous malformation were selected from Department of Neurosurgery, Shenyang General Hospital of Military Area Command of Chinese PLA. There were 12 males and 6 females and their mean age was 40 years. Cerebral arteriovenous malformation was classified based on Spetzler grade: grade Ⅱ (n =10) and grade Ⅲ (n=8). All cases were diagnosed with whole cerebral angiography before operation and they provided the confirmed consent. Main reagents were detailed as follows: endothelial cell growth supplements (ECGS, Sigma, USA), 391 DNA automatic synthetic device (Shanghai Shenggong Liyong Company, PE, USA), anaerobic incubator (DY-1, Zhejiang), human vascular endothelial growth factor enzyme-linked kit (TBD Company, Beijing), 96E enzyme-labeling device (ERMA, INC), cell cycle analytical reagent kit (BD Company), and flow cytometer (FACS Calibur, BD Company).METHODS: ①Experimental procedure: Tissue explants adherent method was used to culture vascular endothelial cells from human cerebral arteriovenous malformation. The third generated cells were used and randomly divided into antisense group, sense group and control group with four bottles of cells in each group. Sense and antisense phosphorothioate oligodeoxynucleotides of artificial vascular endothelial growth factor selected from the antisense group and the sense group were covered with positive liposomes, and then they were used to transfected vascular endothelial cells cultured from human cerebral arteriovenous malformation; however, cells in the control group were not dealt with any treatments. Cells in the three groups were incubated in anaerobic incubator (including 0.95 volume fraction of N2 and 0.05 volume fraction of CO2) at 37 ℃ for 2, 4 and 8 hours, respectively. ② Experimental evaluation: Cell cycles were measured, protein content of vascular endothelial growth factor was measured, and mRNA expression of vascular endothelial growth factor was also detected.MAIN OUTCOME MEASURES: Expression of mRNA and protein of vascular endothelial growth factor and proliferation exponent at different times of hypoxia.RESULTS: ①mRNA expression of vascular endothelial growth factor: At 2, 4 and 8 hours after hypoxia, mRNA expression of vascular endothelial growth factor was higher than that before hypoxia in the control group (P ＜ 0.05);however, mRNA expression was lower in the antisense group than that in the control group (P ＜ 0.05). ② Protein content of vascular endothelial growth factor: At 2, 4 and 8 hours after hypoxia, protein content of vascular endothelial growth factor was higher than that before hypoxia in the control group (P ＜ 0.05); however, protein content was lower in the antisense group than that in the control group (P ＜ 0.05). ③ Proliferation exponent: At 4 and 8 hours after hypoxia,proliferation exponent of endothelial cells cultured from human cerebral arteriovenous malformation was higher than that before hypoxia in the control group (P ＜ 0.05); however, proliferation exponent was lower in the antisense group than that in the control group (P ＜ 0.05).CONCLUSION: Hypoxia may induce gene expression of vascular endothelial growth factor in endothelial cells at the transcriptional level. Antisense vascular endothelial growth factor can obviously inhibit gene expression of vascular endothelial growth factor cultured from human cerebral arteriovenous malformation and proliferation under hypoxic conditions.

Objective To evaluate clinical value of double contrast-enhanced ultrasonography(DCUS)in diagnosing gastric stromal tumors(GST). Methods The medical records of 26 patients with a histological diagnosis of GST were retrospectively reviewed. The correlation between DCUS features and pathological findings of the lesions was compared. Results Total 26 cases of GST were divided into low risk group( 16 cases) and high risk group (10 cases). The size,contour,liquefactive necrosis and enhancement distribution of lesions were markedly correlated with the pathobiologic behaviour of the tumor ( P ＜0.05). There was no significant difference in border and metastasis of lesions between two groups( P ＞0.05). It was shown that DCUS supplied statistically significant improvement in the localization diagnosis and detection of liquefactive necrosis versus oral contrast-enhanced ultrasonography ( P ＜ 0.05 ). Conclusions DCUS is considered as a valuable tool in diagnosing location as well as pathobiological behaviour of GST. It can provide the guidance and reference comments for treatment algorithms.

Objective To investigate the effects of ginkgo flavone on the expression of NF-κB and TLR4 in the liver of mice with nonalcoholic fatty liver disease (NAFLD). Methods 120 KM mice were randomly divided into the control group, model group, as well as high, medium and low dosage of ginkgo flavone groups. The animal model of NAFLD in mice was constructed with high fat diet. The pathological changes of liver, liver index , the serum TNF-α, IL-6 , TG , NF-κB and TLR4 in hepatic tissue was observed after 8 weeks of administration. Results Compared with the model group, the level of liver index, serum TG, TNF-α, IL-6 and the expression of NF-κBp65 in the ginkgo flavone groups dramatically decreased 8 weeks after the administration. And the hepatic steatosis was milder. There was no statistical differences in the expression of TLR4 between the ginkgo flavone groups and the control group (P > 0.05). Conclusions These results suggested the closely relationship between TLR4/NF-κB inflammatory pathway and NAFLD. Ginkgo flavone had the therapeutical effects on NAFLD by anti-inflammatory and lipid-lowering action, but no effect was observed on the expression of TLR4 in hepatic tissue.

Objective To study the human papilloma virus(HPV) infection in lesion tissues of patients with common anus and rectal disease .Methods Gene amplification combined with gene chip technology were employed to conduct genotyping test in lesion tissue of 566 patients with common anus and rectal disease .Results In lesion tissues of 566 patients with common anus and rectal disease ,the overall HPV infection rate was 32 .86% (186/566) .In male patients ,the overall HPV infection rate ,monopole infection rate and multiple infection rate were 32 .14% (117/364) ,23 .35% (85/364) and 8 .79% (32/364) ,respectively ,which showed no sta-tistically significant difference with female [34 .16% (69/202) ,24 .75% (50/202) and 9 .41% (19/202) ,respectively ] (P>0 .05) . HPV 18 ,16 ,33 ,31 types were the main types of common anus and rectal disease .Conclusion HPV genotyping test of anus and rectum tissues is important for molecular epidemiological studies of HPV infection in anus and rectum .

Objective To compare of the effect of intravenous zoledronic acid and calcitonin on the elderly with osteoporotic intertrochanteric fractures.Methods From June 2009 to November 2012,610 patients with osteoporotic intertrochanteric fractures were treated with closed reduction and internal fixation.543 consecutive patients were grouped sequentially according to the admission time.From June 2009 to April 2011,control group (n=325)received calcitonin for osteoporosis (male 107,female 218,mean age 75.02±5.65 years).The number of Evans-Jensen type Ⅰ,Ⅱ,and Ⅲ type patients were 87,136,and 102,respectively.Average lumbar bone mineral density (BMD) and hip BMD were 0.737±0.08 g/cm2 and 0.725±0.05 g/cm2,respectively.From May 2011 to November 2012,218 patients (male 82,female 136,mean age 74.71±5.32 years) received zoledronic acid for osteoporosis.The number of Evans-Jensen type Ⅰ,Ⅱ,and Ⅲ type patient were 62,91 and 65,respectively.Average lumbar BMD and hip BMD were 0.738±0.05 g/cm2 and 0.722±0.06 g/cm2,respectively.All patients received once-yearly intravenous zoledronic acid in one week after operation.The BMD during hospitalization and one year after operation were compared between the two groups.Harris score and VAS score were applied to evaluate the function and pain degree of the operated hip joint.Results The average follow-up time were 12.8 months (range,5 to 22 months) in control group and 12.5 months (range,4 to 19 months) in treatment group.The average fracture union time,Harris score,VAS score were 14.25±1.38 weeks,68.88±5.71 points,and 0.36±0.55 points respectively in treatment group and 14.39± 1.12 weeks,69.47±4.60 points,and 0.33±0.48 points respectively in control group.There were no statistical differences between two groups in fracture union time,Harris score and VAS score.The average lumbar BMD and hip BMD were 0.76±0.06 g/cm2 and 0.75±0.04 g/cm2 in treatment group,which were 0.75±0.07 g/cm2 and 0.74± 0.07 g/cm2 in control group one year after operation,respectively.Compared with the baseline,the BMD of lumbar spine and hip one year after operation slightly increased without statistically significant difference in control group.The BMD increased significantly in treatment group.Conclusion Early administration of intravenous zoledronic acid does not influence bone healing and it could increase BMD one year after closed reduction and internal fixation.