Objective To investigate the expression of E-cadherin(E-cad)and the relationship with the Lauren classification,the degree of histological differentiation,the clinical stage,the depth of invasion,lymph node metastasis and distant metastasis of gastric cancer. Methods 80 cases architecture of gastric cancer and normal tissue were collected.The expression level of E-cad in 80 cases of gastric carcinomas and their metastatic lymph node tissues were examined by immunohistochemical assays.Results The expression rate of E-cad in 80 cases of gastric carcinomas Was 55.00%.The expression level of E-cad was positively correlated with the Lauren classification,the degree of histological differentiation,the clinical stage,the depth of invasion,lymph node metastasis and distant metastasis(P＜0.05),but Was not correlated to patients sex,age and tumor size.The expression rates of E-cad in metastatic gastric carcinoma of primary lesions and lymph nodes metastasis were 35.48%and 32.26%. respectively. Furthermore, E-cad expression in metastatic gastric carcinoma was significandy correlated with primary lesions and lymph node memstasis(r=0.4978,P＜0.05).Conclusion The expression level of E-cad in gastric carcinoma Was closely correlated with the degree of tumor differentiation,infiltration and transferring.

Objective To investigate the effect of platelet-rich plasma (PRP) in anterior cruciate ligament (ACL) reconstruction.Methods From January 2010 to January 2013,40 patients with ACL ruptures who underwent arthroscopic ACL reconstruction with gracilis and semitendinosus tendon were randomly divided into two groups:PRP group and normal saline group.20 patients received graft soaked with PRP and hemocoagulase while 20 patients received graft soaked with normal saline and hemocoagulase.All patients were followed up in 1,3 and 12 months.Evaluation consisted of postoperative drainage volume,inflammatory reaction,grade of wound healed,anterior drawer test,Lachman test,pivot shift,Lysholm knee score and KNEELAX3.Results The average follow-up period was 18 months.Postoperative drainage volume was 142±24 ml in PRP group and 324±22 ml in saline group.The difference was statistically significant.At 4 days after the operation,no inflammatory reaction was observed in 18 cases of PRP group and in 16 cases of saline group,mid inflammatory reaction in 1 case of PRP group and 2 cases of sa line group,and moderate inflammatory reaction in 1 case of PRP group and 2 cases of saline group.Wound healed by first intention in 20 patients of PRP group and in 19 patients of saline group.The preoperative results of anterior drawer test,Lachman test and pivot shift were positive,while postoperative results were negative in both two groups.In PRP group,the preoperative and postoperative Lysholm knee scores of patients in 12 months were 39.8±8.9 and 92.1±2.7 points respectively.In saline group,the preoperative and postoperative Lysholm knee scores of patients in 12 months were 38.7±9.8 and 89.9±4.1points respectively.The differences were not statistically significant.KNEELAX3 measuring results:in PRP group,preoperative measurement was 9.4±1.2 mm in average,while measurement in 12 months postoperatively was 1.2±1.1 mm.In saline group,preoperative measurement was 9.6±1.3 mm,while measurement in 12 months postoperatively was 2.2±1.2 mm.The differences were statistically significant.Conclusion Using graft soaked with PRP in ACL reconstruction could reduce postoperative drainage volume,accelerate the healing of tendon-bone interface in the bone tunnel and the recovery of knee joint function.

Objective To study the changes of serum myeloperoxidase(MPO)in patients with acute ischemic stroke ,and to probe into the relationship of serum MPO with types of carotid atherosclerotic plaques ,the degree of neural function defect and the activi‐ties of daily living (Barthel Index) .Methods Totally 78 cases of patients with acute ischemic stroke was selected as observation ob‐jects .The patients with acute ischemic stroke were divided into good ,medium and poor three groups according to Barthel index . Based on the scoring of neurologic impairment degree from standards of CSS :mild impairment group(0 to 15 points) ,moderate im‐pairment group(16 to 30 points) ,and severe impairment group(31 to 45 points) .Based on the type of atherosclerotic plaques all pa‐tients were divided into soft plaque group ,mixed plaque and hard plaque group .The 1evels of serum MPO was compared between different group .Results The heavier nerve function defect degree ,the levels of serum MPO in patients with ischemic stroke were higher ,and it had significant difference between groups (P<0 .05) .The Barthel index was the better ,the levels of serum MPO was lower .The levels of serum MPO was different among the soft plaque group ,mixed plaque and hard plaque group ,and its were sig‐nificantly different between the three groups ,and the type of atherosclerotic plaque was related to the neural function defect and Barthel index level .Conclusion Ischemic stroke is associated with serum MPO levels ,neurological deficits ,Barthel index and stabil‐ity of atherosclerotic plaque ,and the levels of serum M PO is helpful for judging state of the disease and guiding in clinical diagnosis and treatment .

Objective To investigate the effect of autologous platelet rich plasma (PRP) gel in arthroscopic rotator cuff re?pair. Methods All of 44 patients with rotator cuff tear undergent arthroscopic rotator cuff repair were randomly divided into two groups:PRP group (22 patients were received autologous PRP and hemocoagulase) and normal saline (NS) group (22 patients were received NS and hemocoagulase). All patients had the same accelerated rehabilitation protocol and were followed up in 1, 3, 12 months. Evaluation consisted of inflammatory reaction, wound healed, visual analogue scores (VAS), University of California at An?geles (UCLA) Shoulder Scores and American Shoulder and Elbow Surgeons (ASES) Scores. Results After operation, no inflam?matory reaction was in 20 cases of PRP group and 19 cases of NS group, mid inflammatory reaction 1 case in PRP group and 2 cas?es in NS group, moderate inflammatory reaction 1 case in PRP group and 1 case in NS group. Wound healed by first intention in all of PRP group and 21 patients of NS group. In PRP group, the preoperative, 3 months and 12 months postoperative VAS were 6.6±2.0, 3.4±1.8, 1.8±1.3, UCLA were 15.2±2.9, 24.3±2.7, 32.4±2.1, ASES were 35.6±12.4, 63.4±10.4, 92.3±7.5. In NS group, the preoperative, 3 months and 12 months postoperative VAS were 6.7 ± 1.9, 4.6 ± 1.9, 2.0 ± 1.2, UCLA were 14.8 ± 3.0, 21.2 ± 2.5, 31.7 ± 2.3, ASES were 32.7 ± 13.8, 55.8 ± 11.8, 90.7 ± 8.1. Three months postoperative VAS, UCLA, ASES were statistically signifi?cant differenece in PRP group and NS group. Twelve months postoperative VAS, UCLA, ASES were not statistically significant dif?ferenece in the two groups. Conclusion Using autologous PRP gel in arthroscopic rotator cuff repair can speed up the healing of operation incision with no adverse effect, reduce pain in the postoperation three months, accelerate the rotator cuff repair and re?covery of the function of shoulder joint. It has good short?term clinical effect.

Objective To explore the application of steatosis liver donor (SLD) in piggyback liver transplantation (PBLT). Methods Sixty-four cases of SLD were subjected to PBLT and classified into light steatosis liver (S1,22 cases),moderate steatosis liver (S2,25 cases),and severe steatosis liver (S3,17 cases) groups.Eighty cases of non fatty liver selected randomly in the same period served as controls. The liver and renal function at the day of surgery,postoperative liver function recovery,complications one month after surgery,and the death of recipients were recorded.Results There was no significant difference in the liver and renal function between steatosis liver groups and control group at the day of surgery (P＞0.05). At 21st day after surgery,the liver function of 95％ recipients in control group returned to the normal level,and the liver function recovery rate in S1,S2 and S3 groups was 90.9％,80.0％,and 70.6％ respectively.Graft primary nonfunction occurred in 2 cases (11.8％) of S3 group. The incidence of complications such as bleeding,infection,hepatic artery thrombosis,ascites,sepsis in S1,S2 and S3 groups was higher than in control group (P＜0.05).One year after operation,there were two deaths in control group,one in S1 group,one in S2 group,and 5 in S3 group,respectively.Conclusion SLD can be used for transplantation,but for the transplantation with severe steatosis liver,it should be carried out carefully.

Two kinds of β2-agonistresidues in sheep plasma and urine were disposed by enzymolysis and organic solvent extraction pretreatment methods, and UPLC-MS/MS was used for the qualitative and quantitative analysis. Detection results were compared to study the influences of two pretreatment methods. The experimental results showed that more than 95% of Ractopamine and 40% of Salbutamol exist in the conjugated form in sheep plasma. The detection results of 2 kinds of β2-agonist residues were significantly enhanced when adding β-glucuronidase/aryl sulfatase. The experimental repeatability is very poor ( RSD>40%) when the enzymolysis was not carried out. There were 57% of Ractopamine and less than 1% of Salbutamol exists in the conjugated form in sheep urine. Enzymolysis pretreatment method was useful for the Ractopamine residues determination in urine, and Enzymolysis pretreatment method was useless for Salbutamol determination in urine. Matrix effect of plasma was less than the effects of urine. The influence of organic solvent extraction pretreatment method on the detection results was unremarkable, and there was the possibility that organic solvent extraction could lead partial loss of target compound in extraction process. However, it did not influence the detection results by using internal standard calibration.

A novel method for simultaneous detection of mycotoxins (e.g.,aflatoxin B1) or their metabolic residues in animal plasma with impurity adsorption purification followed ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed.Extraction of mycotoxins and their metabolites from animal plasma sample was performed with 0.1％ formic acid-acetonitrile solution after addition of sodium chloride and hydrous magnesium sulfate.The extract was then dehydrated and purified with hydrous magnesium sulfate,C18,primary secondary amine,and alumina-A.3 mL of the supernatant was evaporated and re-dissolved with 0.5 mL of 0.1％ formic acid aqueous solution/acetonitrile (70∶ 30,V/V) for UPLC-MS/MS detection.The analytes were separated by a C18 column utilizing gradient elution with 0.1％ formic acid aqueous solution containing 0.5 mmol of ammonium acetate and 0.1％ formic acid-methanol solution,and finally detected by tandem mass spectrometry in positive/negative ESI mode.Identification and quantification were achieved by LC-MS/MS with multi-reaction monitoring (MRM).Good linearity in response was obtained in the analytes concentration range of 0.05-100 ng/mL with correlation coefficients larger than 0.99.The limits of quantification (S/N=10) were around 0.05-0.5 ng/mL.The recoveries of mycotoxins and their metabolites spiked in blank plasma samples were in the range of 62.0％-116.4％,with relative standard deviations (RSDs) less than 19.0％.

A rapid high-throughput method for the determination of 26 mycotoxins involving multifunctional cleanup column coupled with liquid chromatography-tandem mass spectrometry ( LC-MS/MS) was developed and validated for the determination in feedstuffs. The feedstuff samples were extracted by ultrasonic treatment for 1 hour and the extraction solvent was acetonitrile/water/formic acid (84:15. 9:0. 1, V/V). 1 mL of the supernatant layer was purified by a commercial Mycospin 400 multifunctional cleanup column, then dried and re-dissolved by 0. 25 mL water/methanol/formic acid (95:4. 9:0. 1, V/V) in a vial for injection into the LC-MS/MS system. Chromatographic analyses were carried out on a reversed phase C18 column and using a gradient elution with 0. 1% formic acid aqueous solution and 0. 1% formic acid methanol solution. The mass spectrometer was operated in a multiple reaction monitoring ( MRM) mode that selected one precursor ion and two product ions for each target compound. Validation studies were carried out in maize and soybean meal as representative matrixes. The most target compounds had different level of matrix effects. So, matrix-matched calibration was adopted for quantification. Mean recoveries from spiked samples at three levels ranged from 61 . 9% to 119 . 5% with relative standard deviations of 0 . 8%-18 . 6%. Limits of quantification ranged from 0. 5 μg/kg to 25 μg/kg.

Objective To observe the changes of TNF-α and NF-κB after different doses of nalmefene hydrochloride (NAL) therapy for traumatic brain injury (TBI) in an effort to identify the effect of NAL on TBI-induced inflammatory response and the possible mechanism.Methods A model of TBI in the rat was produced using the improved Feeney' s free-fall impact method.The animals were randomly divided into sham group,TBI group,TBI + large dose of NAL (ip,0.2 mg/kg) group (TBI + NAL1group),TBI + medial dose of NAL (ip,0.14 mg/kg) group (TBI + NAL2 group),TBI + small dose of NAL (ip,0.07 mg/kg) group (TBI + NAL3 group).Form of brain tissues in each group was observed and mRNA levels of TNF-α and NF-κB were measured by real-time quantitative PCR assay.Results HE staining revealed severe injury and inflammatory infiltration of brain parenchyma in TBI group ;on the contrary,the situation ameliorated in TBI + NAL1 group,TBI + NAL2 group and TBI + NAL3group,with especially obvious improvement in TBI + NAL2 group.In PCR assay,significant expression of NF-κB and TNF-α was observed at post-TBI days 1,3,5 and 7 (P ＜ 0.05),followed by great reverse after NAL therapy (P ＜ 0.05),particularly in TBI + NAL2 group.Conclusions NAL can reduce the inflammation response to TBI and promote post-injury recovery.Moreover,there exists a NAL concentration window.

Objective To establish a direct reverse transcription real-time fluorescence quantitative polymerase chain reaction ( RT-qPCR-D ) method for detecting serum circulating B cell-specific moloney murine leukemia virus integration site-1 (Bmi-1) mRNA, and analyze the levels of serum circulating Bmi-1 mRNA in colorectal cancer patients by using of this method for exploring its diagnosis value in colorectal cancer.Methods Methodology establishment.RNA was extracted from colorectal cancer HT 29 cell line, and detection standard curves of Bmi-1, ubiquitin C ( UBC), glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) mRNAs were established , then the amplification efficiencies were calculated.Bmi-1 mRNA level was directly detected in serum and preparation buffer mixture , then the specificity of assay was evaluated by melting curve, and detection limit was observed through diluted serum samples.The serum circulating Bmi-1 mRNA levels were detected by ELISA in 158 cases with colorectal cancer , of which there were 26 cases of tumor node metastasis ( TNM)Ⅰstage, 53 cases of TNMⅡ, 47 cases of TNMⅢ, 32 cases of TNMⅣand 53 cases of controls with normal colonoscopy collected from January 2008 to January 2009 in Qilu Hospital of Shandong University.Comparisons of groups were determined by applying Mann-Whitney U test or Kruskal-Wallis test, and receiver operating characteristic ( ROC) curves were established to illustrate the diagnostic performance.Results The log values of Bmi-1, UBC and GAPDH showed good linear correlations with quantification cycle (Cq) values(R2 =0.990, 0.990, 0.991, all P <0.001), and the amplification efficiencies were 0.875, 0.917 and 0.935, respectively.Using the established RT-qPCR-D method, the peak of melting curve of Bmi-1, UBC and GAPDH mRNAs were single, the detection limit was up to 1.25μl.The levels of serum circulating Bmi-1 mRNA detected by RT-qPCR-D were 0.138 ( 0.078-0.228 ) in colorectal cancer stage Ⅰ patients, 0.163(0.067 -0.287) instage Ⅱ patients, 0.217(0.072-0.267) instage Ⅲpatients, 0.273(0.139 -0.419) in stage Ⅳ patients and 0.021(0.008 -0.029) in health controls, a significant difference was found among groups ( H =89.5, P <0.001 ).The levels of serum circulating Bmi-1 mRNA in each stage colorectal cancer were all significantly higher than that in control group(U=58.0, 287, 246, 72.5,all P<0.001).The levels in Ⅳstage patients were significantly higher than those in other stages patients (U=247, 590, 540,P=0.008, 0.020, 0.035), while no significant differences among Ⅰstage,Ⅱstage and Ⅲstage patients(U=633, 514, 1170,all P>0.05).ROC curve analysis showed area under the ROC curve ( AUC) for serum circulating Bmi-1 mRNA was 0.921(95%CI=0.876-0.953), which was significantly superior to the AUC of CEA (0.745, 95%CI=0.680-0.802, Z=4.697, P<0.001 ).When cutoff value was 0.034, the diagnostic sensitivity and specificity was 89.2%(141/158) and 90.6%(48/53), while 41.8%(66/158) and 73.6%(39/53) using CEA.The AUC for combination of circulating Bmi-1 mRNA and CEA was 0.933(95%CI=0.890-0.963 ) , which was no statistical significance when compared with the AUC of circulating Bmi-1 mRNA(Z=4.697, P>0.05).Conclusions The study establishes a higher sensitive, specific for detecting serum circulating Bmi-1 mRNA. Based on this method , serum circulating Bmi-1 mRNA is found to be increased in colorectal cancer , and is superior to traditional tumor marker CEA in diagnosis of colorectal cancer, which may become a potential detection index for early detection of colorectal cancer.