Objective To prepare biomimetic bone material by reconstructing type Ⅰ collagen combined with polyaspartic acid. Methods By acid hydrolysis,rat tail type Ⅰ collagen was decomposed into collagen fibers,which were then placed in the calcium phosphate mineralization solution. Under the cross-linking of glutaraldehyde,the collagen fibers were reconstructed and assembled into collagen fibers,and the calcium phosphate crystals were wrapped in the inner side of the collagen fibers for biomineralizationin. After poly aspartate acid was added,calcium hydroxyapatite calcium precursor was added into the collagen fibers to simulate thebiomimetic biomineralizationin the human body. After mineralization for 3-9 days,the bone mineralization process was observed by transmission electron microscopy and electron diffraction. Results Transmission electron microscopy and electron diffraction displayed that,after 3 days of mineralization,calcium hydroxyapatite precursor was wrapped in the collagen fiber gap,and the collagen fiber was partially mineralized. After 9 days of mineralization,calcium hydroxyapatite precursor completely infiltrated into the collagen fiber,and the amorphous calcium phosphate was transformed into hydroxyapatite calcium crystal. Thus,the simulation of bone mineralization was completed,and collagen type Ⅰ collagen/hydroxyapatite calcium biomimetic bone material was formed. Conclusion Reconstruction of type Ⅰ collagen combined with polyaspartic acid can prepare biomimetic bone material that has close chemical composition and molecular structure to the human bone tissue.

Objective To explore the curative efficacy of treating tibial transport gap fracture after bone transport by external fixation with locking compression plate(LCP) and autologous iliac grafting.Methods From February 2015 to January 2016,9 patients who had sustained tibial transport gap fracture after bone transport were treated by LCP external fixation and autologous iliac grafting.They were 7 men and 2 women,aged from 26 to 56 years (average,40.2 years).One of them received bone transport because of limb shortness after replantation and others did because of traumatic osteomyelitis.The distances of tibial transport averaged 9.2 cm (from 7 to 12 cm);the time for external fixation averaged 20.1 months (from 13 to 25 months);the time from removal of external fixator to gap fracture averaged 1.8 weeks (from 1 to 3 weeks).Two patients were complicated with docking site fracture.The durations from gap fracture to operation averaged 4.1 days (from 3 to 5 days).Five patients sustained angular deformity of various severities which could not be corrected by surgery.The curative efficacy was evaluated according to conventional criteria for fracture healing.Results The 9 patients were followed up for 11 to 15 months (average,13.1 months).The time for LCP external fixation averaged 9.0 months (from 8 to 10 months);the time for fracture union averaged 4.6 months (from 4 to 5 months).The 5 patients with angular deformity obtained fracture malunion which did not obviously affect their limb appearance.One case suffered extensive cellulitis at the leg which responded to intravenous administration of antibiotics.No pin track infection happened.The knee and ankle functions after removal of LCP external fixation were not significantly different from those after removal of external fixator following bone transport.Wounds at the iliac donor site and bone graft area all healed well.Conclusions LCP external fixation is an effective treatment for tibial transport gap fracture after bone transport,due to its stable fixation,limited injury to soft tissues,positive curative efficacy and miniature size as well.However,it requires sophisticated operative skills and demanding postoperative care.

OBJECTIVE:To establish the method for the determination of plasma concentration of magnesium isoglycyrrhiz-inate in portal vein and peripheral venous blood of patients underwent liver resection,to further validate and evaluate pharmacoki-netic characteristics,rational and safe use of drugs in the clinic. METHODS:31 patients underwent liver resection in our hospital during Oct. 2014-Mar. 2015 were given magnesium isoglycyrrhizinate intravenously at the beginning of surgery. Portal vein and pe-ripheral venous blood of patients were drawn at 1 hour after drug use,and HPLC-UV detection method was used to determine the plasma concentration of drug. RESULTS:The retention time of isoglycyrrhizinate magnesium was 4.5 min,which showed a good peak shape,and was not interfered with the determination by plasma endogenous peak. The plasma concentration ranged from 0.55 to 55.00 mg/L. The minimum quantitative concentration was 0.55 mg/L. The extraction recoveries were 84.7%-87.1%,and method recoveries were 101.2%-105.4%,and RSDs of intra-day and inter-day were less than 6%. Plasma concentration of magnesium iso-glycyrrhizinate in portal vein blood was significantly higher than in peripheral vein blood of patients underwent liver resection (close to 2 times);and plasma concentration was not affected by primary liver diseases and underlying diseases such as cirrhosis. CONCLUSIONS:The method is simple and has high recovery rate of extraction,high accuracy and high sensitivity. It can meet the needs of pharmacokinetic study. After the application of magnesium isoglycyrrhizinate during liver resection,there is higher blood concentration of magnesium isoglycyrrhizinate in portal vein,which is beneficial to protect liver cells and improve liver func-tion. It is suitable during perioperative period of liver.

Objective To investigate the association between body mass index (BMI), waist circumference (WC), waist?to?height ratio (WHtR), and the incidence risk of type 2 diabetes mellitus (T2DM). Methods In total, 20 194 participants≥18 years old were selected randomly by cluster sampling from two township (town) of the county in Henan province from July to August of 2007 and July to August of 2008 and the investigation included questionnaires, anthropometric measurements, fasting plasma glucose,and lipid profile examination were performed at baseline; 17 236 participants were enrolled in this cohort study. 14 720 (85.4%) were followed up from July to August 2013 and July to October 2014. Finally, 11 643 participants (4 301 males and 7 342 females) were included in this study. Incidence density and Cox proportional hazards regression models were used to evaluate the risk of T2DM associated with baseline BMI, WC, WHtR, and their dynamic changes. Results After average of 6.01 years following up for 11 643 participants, 613 developed T2DM and the incidence density was 0.89 per 100 person?years. After adjusted for baseline sex, age, smoking, drinking, family history of diabetes, as well as the difference of fasting plasma?glucose (FPG), total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL?C), systolic blood pressure (SBP), diastolic blood pressure (DBP) between baseline and follow?up, Cox Proportional?Hazards regression analysis indicated that T2DM risk of baseline BMI overweight group, BMI obesity group, abnormal WC group and abnormal WHtR group were significantly higher than that of the corresponding baseline normal groups , and the incidence risk of T2DM reached the highest for those whose baseline BMI, WC and WHtR were all abnormal, the corresponding HR (95%CI) were 2.05 (1.62-2.59), 3.01 (2.33-3.90), 2.34 (1.89-2.90), 2.88 (2.21-3.74), 3.32 (2.50-4.40), respectively. Whether baseline BMI/WC was normal or not, T2DM risk increased if baseline WHtR was abnormal, and the HR (95%CI) of baseline normal BMI/abnormal WHtR group, baseline abnormal BMI/abnormal WHtR group, baseline normal WC/abnormal WHtR group, baseline abnormal WC/abnormal WHtR group were 1.88 (1.29-2.74), 3.08 (2.34-4.05), 2.15 (1.53-3.00), 3.22 (2.45-4.23), respectively. The analysis for dynamic changes of BMI, WC, and WHtR indicated that in baseline normal WC or WHtR group, T2DM risk increased when baseline normal WC or WHtR developed abnormal at follow?up, and the corresponding HR (95%CI) were 1.79 (1.26-2.55), 2.12 (1.32-3.39), respectively. In baseline abnormal WC or WHtR group, T2DM risk decresed when baseline abnormal WC or WHtR reversed to normal at follow?up, and the corresponding HR (95%CI) were 2.16 (1.42-3.29), 2.62 (1.63-4.20), respectively. Conclusion BMI, WC, and WHtR were associated with increased T2DM risk. The more abnormal aggregation of BMI, WC, and WHtR presents, the higher T2DM risk was. T2DM risk could be decreased when abnormal WC or WHtR reversed to normal.

Objective To investigate the association between body mass index (BMI), waist circumference (WC), waist?to?height ratio (WHtR), and the incidence risk of type 2 diabetes mellitus (T2DM). Methods In total, 20 194 participants≥18 years old were selected randomly by cluster sampling from two township (town) of the county in Henan province from July to August of 2007 and July to August of 2008 and the investigation included questionnaires, anthropometric measurements, fasting plasma glucose,and lipid profile examination were performed at baseline; 17 236 participants were enrolled in this cohort study. 14 720 (85.4%) were followed up from July to August 2013 and July to October 2014. Finally, 11 643 participants (4 301 males and 7 342 females) were included in this study. Incidence density and Cox proportional hazards regression models were used to evaluate the risk of T2DM associated with baseline BMI, WC, WHtR, and their dynamic changes. Results After average of 6.01 years following up for 11 643 participants, 613 developed T2DM and the incidence density was 0.89 per 100 person?years. After adjusted for baseline sex, age, smoking, drinking, family history of diabetes, as well as the difference of fasting plasma?glucose (FPG), total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL?C), systolic blood pressure (SBP), diastolic blood pressure (DBP) between baseline and follow?up, Cox Proportional?Hazards regression analysis indicated that T2DM risk of baseline BMI overweight group, BMI obesity group, abnormal WC group and abnormal WHtR group were significantly higher than that of the corresponding baseline normal groups , and the incidence risk of T2DM reached the highest for those whose baseline BMI, WC and WHtR were all abnormal, the corresponding HR (95%CI) were 2.05 (1.62-2.59), 3.01 (2.33-3.90), 2.34 (1.89-2.90), 2.88 (2.21-3.74), 3.32 (2.50-4.40), respectively. Whether baseline BMI/WC was normal or not, T2DM risk increased if baseline WHtR was abnormal, and the HR (95%CI) of baseline normal BMI/abnormal WHtR group, baseline abnormal BMI/abnormal WHtR group, baseline normal WC/abnormal WHtR group, baseline abnormal WC/abnormal WHtR group were 1.88 (1.29-2.74), 3.08 (2.34-4.05), 2.15 (1.53-3.00), 3.22 (2.45-4.23), respectively. The analysis for dynamic changes of BMI, WC, and WHtR indicated that in baseline normal WC or WHtR group, T2DM risk increased when baseline normal WC or WHtR developed abnormal at follow?up, and the corresponding HR (95%CI) were 1.79 (1.26-2.55), 2.12 (1.32-3.39), respectively. In baseline abnormal WC or WHtR group, T2DM risk decresed when baseline abnormal WC or WHtR reversed to normal at follow?up, and the corresponding HR (95%CI) were 2.16 (1.42-3.29), 2.62 (1.63-4.20), respectively. Conclusion BMI, WC, and WHtR were associated with increased T2DM risk. The more abnormal aggregation of BMI, WC, and WHtR presents, the higher T2DM risk was. T2DM risk could be decreased when abnormal WC or WHtR reversed to normal.

ObjectiveTo explore the surgical treatments and the efficacy of hilar cholangiocarcinoma.MethodsClinical data of 113 patients with hilar cholangiocarcinoma in District of Huangdao, the Affiliated Hospital of Medical College Qingdao University from June 1999 to June 2009 were studied retrospectively. The informed consents of all patients were obtained and the ethical committee approval had been received. There were 55 males and 58 females with age ranging from 37 to 70 years old and a median age of 61 years old. After admitted in hospital, the patients underwent a comprehensive medical examination and received routine tests of liver function, tumor markers and imaging examinations. The surgical treatments included radical resection, palliative resection, internal drainage and external drainage. The patients were followed up after operation. The operation, death and incidence of complications were observed. Survival analysis were conducted using Kaplan-Meier method and Log-rank test.ResultsThere were 22 cases with hilar cholangiocarcinoma of Bismuth-Corlette typeⅠ, 13 cases typeⅡ, 21 cases typeⅢa, 18 cases typeⅢb, 39 cases typeⅣ. The surgical resection rate was 72.6% (82/113), including radical resection (n=53), palliative resection (n=29), internal drainage (n=19), external drainage (n=12). Three cases died after operation, including 2 cases died of liver failure, 1 case died of upper gastrointestinal bleeding. After operation, upper gastrointestinal bleeding were observed in 3 cases, hepatic encephalopathy in 5 cases, bile leakage in 18 cases, abdominal infection in 3 cases, pleural effusion in 11 cases, wound infection and dehiscence in 7 cases. The 1-, 3-, 5-year cumulative survival rate were 76%, 39%, 11% respectively for patients of radical resection, 74%, 21%, 4% for palliative resection, 35%, 2%, 0 for internal drainage, and 28%, 0, 0 for external drainage. There were significant differences in the survival rates for patients of 4 surgical treatments (χ2=21.367,P<0.05).ConclusionImprovement of preoperative evaluation and selection of appropriate surgical approach help to improve the outcome and survival rate of patients with hilar cholangiocarcinoma.

ObjectiveTo study the effect of α7 ( α7 AChR) agonist nicotine on regulating sensitivity of regular chemotherapeutic agent in cholangiocarcinoma cells,and explore the possible target.MethodsThe effect of nicotine and α-BTX pretreatment on the survival ability of cholangiocarcinoma cells was investigated when applied with 5-FU by using MTT and Flat cloning formation experiment.ResultsApplied with 5-FU,in various con centrations nicotine stimulating group( 10-3 g/L,10-4 g/L,10-5 g/L ),the survive rate of QBC939 was 128％,124％,118％,while that in α-BTX stimulating group and combined stimulation group was 92％,94％,93％,92％,respectively.The cloning formation ability of nicotine- stimulating group (6.2 ± 0.40) was significantly higher than α- BTX stimulating group (3.2 ± 0.20 ),combined stimulation group ( 3.2 ± 0.20 ) and control group ( 3.4 ±0.33).ConclusionNicotine can prevent chemotherapy-induced apoptosis,and improve cholangiocarcinoma cell survival via α7 nicotine acetylcholine receptor in vitro.

Objective To explore the experiment condition and method for the application of in vitro in vasive Transwell chamber and to observe muscarinicreceptor stimulant and muscarinicreceptor antagonist's influence to cholangiocarcinoma's invasiveness.Methods Two hundred microliter cell suspension of various concentrations(0.5×105/mL,1.0×105/mL,1.5×105/mL and 2.0×105/mL)was added into the upper chamber of the Transwell chamber,and the cells were allowed to penetrate the matrigel for 12,18,24and 48 hours respectively.The numbers was gotten as the invasive cells on the under surface of the membrane.After optimal cell concentration and time were gotten,pilocarpine of various concentrations(0 mmol/L,0.1 mmol/L,0.3 mmol/L and 0.5 mmoL/L)was added into the upper chamber of the Transwell chamber,then the cells on the matrigel were stained and counted.So did the cells when atropine of various concentrations(0.01 mmol/L,0.01 mmol/L,0.05 mmoVL and 0.1 mmol/L)were added into the upper chamher of the Transwell chamber in according to pilocarpine of various concentrations(0 mmol/L,0.3 mmol/L,0.3 mmol/L and 0.3mmol/L).Results With the increase of the time and cell concentrations，the cells couts that penetrated the matrigel increased，while the increase tended to he stable when the culture time exceeded 36 hous and the cell concentration Was over 1.0×105/mL.By adding pilocarpine,there were significant differences between the control and experimental groups(P＜0.05)，but there were no significant differences in experimental groups with various concentrations.There were no significant differences in blank group and experimental groups with atropine added(P＞0.05).When added pilocarpine and atropine,there were significant differences between blank and experimental groups(P＜0.05)，but there were no significant differences in experimental groups with various concentrations.Conclusions Thirty-six hours as invasive time,and one cell concentration 1.0 × 105/mL were optimal to test invasion abilities of cholangiocarcingma cells to different medicines or reagents.There is the possibility that museariniereceptor exists in cholangiocarcinoma cells，and may play an important role in cholangiocarcinoma's invasiveness and metastasis.

Objective To investigate expression of slug and E-cadherin in pancreatic cancer tissues and determine the inhibitory effects of anti-Slug, an anti-sense plasmid, on the invasion of pancreatic cancer cell lines in vitro. Methods Slug and E-cadherin protein and mRNA was analyzed by IHP and RT-PCR in 36 cases of pancreatic cancer. Then anti-Slug plasmid was transfected into herin and Slug expression. The inhibitory effects of anti-sense Slug were also detected by Transwell motility assay and Matrigel invasion assay. Results The expression of Slug and mRNA in metastatic pancreatic cancer tissue was higher than that in non-metastatic tissue. E-cadherin and mRNA was lower in metastasis tissues(P＜0.05). The inverse relationships were further observed by transient transfection of anti-Slug into SW1990H4 cells. The downregulated expression of Slug and re-expression of E-cadherin were found. The Slug mRNA levels were 0.985±0.016,0.973±0.014, 0. 554±0. 011 after 0, 48 h of transfection of anti-sense Slug, and that of E-cadherin were 0.120±0.001, 0.360±0.002, 0. 727±0. 006, respectively. The diference was significant between different time points (P＜0.05). The Slug mRNA levels were 0. 206±0.017, 0.968±0.015, and that of E-cadherin were 0. 18±0.002,0.727±0.006 after stable transfection of anti-sense Slug, and control plasmid, respectively. The diference was significant (P＜0.05). The motility activity(393±28, 352±24, 96 ±13 )and the invasion activity (223 ± 69, 202 ± 64, 65 ±19) of1 antisense Slug transfectant cells were significantly decreased as compared with those of control cells (P＜0.05). Conclusions Higher expression of slug and lower expression of E-cadherin is related to the invasion and metastasis in pancreatic cancer. A reverse corelation of E-cadherin and Slug expression exists in pancreatic cancer. Slug is possibly a potential target for cancer gene therapy blocking invasion and metastasis in human pancreatic cancer.

Cholangiocarcinoma is a type of malignant tumor with high destruction.Due to its low diagnostic rate and high fatality rate,the operation is the unique therapeutic methods for the radical cure.However,the diagnosis and treatment for the disease were always in the phase of progression,so currently,the radical therapeutic rate is quite low,while the recurrence rate of the operation is extremely high.If the correlated mechanism of perineural invasion of cholangiocarcinoma could be understood,then interrupted its perineural invasion in the early period,that could greatly enhance the prognosis of cholangiocarcinoma patients.This article systematically reviews the progress of cholangiocarcinoma neural invasion related molecules and possible mechanism.