Objective To investigate the diagnostic value of a quantitative detection of K-ras mutation in samples from endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA)of pancreatic cancer.Methods Samples taken by EUS-FNA from 53 pancreatic occupying lesions were collected, and the copies of wild-type and mutated K-ras gene was measured by PNA-clamping real-time quantitative PCR. The results were analyzed with refer to cytological findings to evaluate its clinical values. Results According to cytological finding, a total of 37 cases were diagnosed as pancreatic cancer, and 16 were non-malignant lesions. Kras mutation was detected in 83.8% of cancer cases, and 18. 8% of non-cancer cases, which was significantly different ( P ＜0. 05 ). Sensitivities of cytology and K-ras examination were 59. 5% and 83.8%, respectively, while that of combination of cytology and K-ras examination was 89. 2%. Conclusion Quantitative analysis of the mutant K-ras gene in samples taken by EUS-FNA is a useful tool for diagnosing the pancreatic carcinoma.

Objective To investigate whether tissue engineered bone with implanted vascular bun-dles can up-regulate release of vascular endothelial growth factor (VEGF) in models of femoral defect in rabbits.Methods Thirty-two rabbits were randomized into 2 even groups.In both groups, a segmental bone defect of 15 mm in length was made at the left femur before a tissue engineered bone was inserted into the defect.In the experimental group, a femoral vascular bundle was implanted into the tissue engineered bone.In the control group, there was no vascular implantation.At 2, 4, 8, and 12 weeks after implantation, samples were taken to determine new bone formation by histology and expression level of VEGF by immuno-histochemistry.Results The new bone formation was significantly higher in the experimental group at the end of 4, 8, and 12 weeks(P < 0.05) .The expression level of VEGF in the experimental group was also significantly higher than in the control group at all time points after operation, and the expression of VEGF peaked at 4 weeks.Conclusion Tissue engineered bone with vascular bundle implanted can up-regulate VEGF release in models of femoral defect in rabbits.

ObjectiveTo detect the Alu expression in the stool of patients with pancreatic cancer and investigate its value in the diagnosis of pancreatic cancer.MethodsStool samples were obtained from patients with pancreatic cancer (PC) ( n =41 ),chronic pancreatitis (CP) ( n =27 ) and healthy subjects ( n =23 ),the DNA was extracted from the stool and the expression of Alu repetitive sequences was subjected to quantitative analysis by the real-time PCR.ResultsThe expressions of Alu repetitive sequences in PC,CP,and healthy subjects were (5.17 ± 0.99 ),( 3.79 ± 0.94),(0.28 ± 0.35 ) rig/g,and the difference among the three groups was statistically significant (P ＜0.05).The AUC of PC was 74.8％ with the 95％ CI 0.661 ～0.835,and the sensitivity,specificity was 75.6％ and 67.1％,respectively.ConclusionsAlu repetitive sequences are highly expressed in the stool of patients with pancreatic cancer,and it is of value in the diagnosis of pancreatic cancer.

Objective To explore whether the respective implantation of vascular bundles and sensory nerve tracts into a tissue-engineered bone will affect the expression of CGRP (Calcitonin gene related peptide) and its receptor. Methods Fifty-four New Zealand rabbits were randomly divided into 3 even groups for implantation of sensory nerve tracts (group A),implantation of vascular bundles (group B),and a control group of simple tissue-engineered bone (group C) . Animals were sacrificed 4,8,12 weeks after implantation,respectively. Masson staining was conducted to observe the process of bone formation and re-molding. CGRP and CGRPR-1 expressions in the new bone were measured by immunohistochemistry and Real-time PCR at 4,8 and 12 weeks after implantation. Results At all time points,the CGRP and CGRPR-1 expressions in groups A and B were significantly higher than in group C (P<0.05),and those in group A were higher than in group B too (P<0.05) . Over time,the expressions of CGRP and CGRPR-1 mRNA in each group in the new bone tissue were gradually reduced after an initial increase. The neuropeptide expression at the 8th week was higher than those at the 4th and 12th weeks. The neuropeptide expression at the 4th week was the lowest. The expression of CGRP was mainly localized in the periphery of newly generated bone,periosteum and the blood vessels. The expression of CGRPR-1 was mainly localized in the periphery of osteoblasts. Conclusions Implantation of either vascular bundles or sensory nerve tracts can promote neuropeptide secretion. The vascular bundle implantation may result in higher expressions of CGRP and CGRPR-1 than sensory nerve tract implantation.

Objective To explore characteristics and significance of the indexes of peripheral white blood cell (WBC) in patient with human brucellosis.Methods People checked by brucellosis physical checkup and routine physical checkup at Qiqihar Center for Disease Control and Prevention from December 2014 to December 2015,including 40 acute brucellosis patients (acute group),35 chronic brucellosis patients (chronic group) and 72 healthy people (control group),were selected.Automatic blood analyzer was used to determine the indexes of WBC,lymphocyte count (LY),lymphocyte percentage (LY％),monocytes count (MONO),monocytes percentage (MONO％),eosinophil count (EO),eosinophil percentage (EO％),basophilic granulocyte count (BASO),basophilic granulocyte percentage (BASO％),neutrophils count (NEUT) and neutrophils percentage (NEUT％).The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of WBC parameters in acute and chronic groups.Results Compared to control group,the levels of WBC,EO,EO％,BASO,BASO％,NEUT and NEUT％ were decreased in acute group [(5.222 0-± 2.551 2) × 109/L vs (6.352 5 ± 1.905 8) × 109/L,(0.030 0 ± 0.006 8) × 109/[,vs (0.083 9 ± 0.039 3) × 109/L,(0.54 ± 0.12)％ vs (2.31 ± 0.14)％,(0.009 0 ± 0.001 1) × 109/L vs (0.019 0 ± 0.002 4) × 109/L,(0.17 ± 0.09)％ vs (0.32 ± 0.20)％,(2.698 7 ± 1.948 4) × 109/L vs (4.012 9 ± 1.579 0) × 109/L,(48.13 ± 14.38)％ vs (62.13 ± 9.00)％,all P ＜ 0.05],and the levels of LY,LY％ and MONO％ were increased in acute group [(2.125 3 ± 0.949 9) × 109/L vs (1.794 4 ± 0.606 6) × 109/L,(43.37 ± 14.52)％ vs (29.10 ± 7.97)％,(7.84 ± 2.23)％ vs (6.55 ± 2.04)％,all P ＜ 0.05].Compared to control group,the level of NEUT％ [(54.63 ± 9.26)％] was decreased in chronic group (P ＜ 0.05),and the levels of LY,LY％ and EO [(2.212 0 ± 0.633 2) × 109/L,(36.41 ± 8.51)％,(0.153 9 ± 0.028 8) × 109/L] were increased in chronic group (all P ＜ 0.05).The levels of LY％ and MONO％ [(6.45 ± 1.58)％] in chronic group were lower than those in acute group (all P ＜ 0.05),and the levels of WBC [(6.175 7 ± 1.469 5) × 109/L],EO,EO％ [(2.32 ± 1.21)％],BASO [(0.021 8 ± 0.001 9) × 109/L],BASO％ [(0.37 ± 0.21)％] and NEUT％ were higher than those in acute group (all P ＜ 0.05).The areas under ROC curve (AUCs) of LY and MONO in acute group were 0.681 and 0.529,they were in 0.5-0.7,and the diagnostic value was low;the AUCs of EO,EO％,LY％,NEUT％,NEUT,BASO,BASO％,MONO％ and WBC in acute group were 0.816,0.816,0.806,0.790,0.766,0.760, 0.721,0.715 and 0.710,they were in ＞ 0.7-0.9,and the diagnostic value was medium.The AUCs of LY,NEUT,BASO,EO,BASO％,EO％,MONO％,MONO and WBC in chronic group were 0.693,0.617,0.586,0.584,0.581,0.541,0.500,0.513 and 0.510,they were in 0.5-0.7,and the diagnostic value was low;the AUCs of LY％ and NEUT％ in chronic group were 0.725 and 0.717,they were in ＞ 0.7-0.9,and the diagnostic value was medium.Conclusion The indexes of peripheral WBC in patient with acute and chronic human brucellosis are changed abnormally,which has a certain reference value in diagnosis of human brucellosis.

Objective To detect the microRNAs in fecal with patients of pancreatic cancer, and evaluate its diagnostic value. Methods Stool samples were collected from three group persons including 29 pancreatic cancer, 22 chronic pancreatitis and 13 normal controls. The total fecal microRNAs were extracted.The quantity of miR-16, miR-21, miR-155, miR-181a, miR-181b, miR-196a, and miR-210 were detected by using real-time PCR, and miR-16 was used as reference gene. ROC AUC was used to evaluate the diagnostic value for pancreatic cancer. Results MicroRNAs were efficiently obtained from stools, and independent experiments showed high reproducibility for microRNAs extraction and detection. The expression of miR-181b,miR-196a, miR-210 in fecal was 2.22 ±0.64,2.78 ±0.14, 5.55 ±0.38 in pancreatic cancer; 1.42 ±0.39,3.88 ± 0.85,5.39 ± 0.69 in chronic pancreatitis; 0.32 ± 0.40, 1.14 ± 0.98,4.23 ± 0. 99 in normal controls;the three microRNA expressions in pancreatic cancer were group and CP group significantly higher than those in normal controls ( P ＜ 0.05 ). But there was no significant difference between pancreatic cancer group and chronic pancreatitis group. AUC of pancreatic cancer / normal controls miR 18lb was 0.745(95% CI 0. 597-0.894), the sentivity, specificity for pancreatic cancer was 84.6% and 51.7%. AUC of miR-210 was 0. 772(95% CI0.629-0.914), the sentivity, specificity for pancreatic cancer was 84.6% and 65.5%, and the difference was statistically significant (P ＜0.05). miR-196a was no significant for the diagnosis of pancreatic cancer, but the expression of miR-196a was correlated with the tumor size (r = 0.516, P = 0.041 ).Conclusions The extraction and detection of the fecal microRNAs were non-invasive and reproducible. The expression of miR-181b and miR-210 was increased in stool of patients with pancreatic cancer, and may be potential biomarker for pancreatic cancer.

Objective To investigate the diagnostic value of the K-ras mutations in FNA samples for early detection of pancreatic cancer. Methods FNA samples of 27 patients with pancreatic cancers, 9 patients with other malignant tumors and 14 patients with non malignant pancreatic mass (NMPM) were collected. DNA was extracted, and K-ras gene was amplified through PNA-mediated PGR clamping, the products were sequenced to determine the mutation type. Results The positive rate of K-ras mutations in pancreatic cancers,other malignant tumors and NMPM were 88.9%, 44.4%, 35.7%. There was significant difference in K-ras gene mutations in FNA samples between pancreatic cancer and other malignant tumors ( P = 0. 013 ) and NMPM ( P = 0. 001 ). The sensitivity, specificity, positive predictive value, negative predictive value,accuracy of K-ras mutations in FNA samples of pancreatic cancers were 88.9%, 55.6%, 85.7%, 62.5%,80.6% when compared with other malignant tumors, and the difference between the two groups was significant (P =0. 013) ;Those were 88.9%, 64.3%, 82.8%, 75.0%, 80. 5% when compared with NMPM, and the difference between the two groups was significant ( P = 0. 001 ). When cytology of FNA samples and K-ras mutations was combined, the positive rate of pancreatic cancer was up to 96.3%. Conclusions The detection of K-ras mutations in EUS-FNA samples helped improve the positive diagnostic rate of pancreatic cancer.

Objective: To investigate the effect of free abdominal flap in repairing soft tissue defects of the foot. Methods: From 2011.10 to 2017.11, there were 5 patients whose foot had soft tissue defect, in order to repairing, we had to do debridment at the early time, then made the anterior iliac artery or the inferior epigastric artery as the axial vessel. Results: After the flap transplantation there was no vascular crisis in the 5 patients , followed up for 3 months to 1 years, the blood supply, texture, elasticity and the foot was well in shape and feeling, the patients were satisfied, and the feet were functionally normal. Conclusions The application of free abdominal skin flap covers the soft tissue defect of the foot, the wound surface, the blood vessel the donor site is consistent, the flap is easy to prepare, and the area can be sutured directly, this is an effective treatment method.

Objective To investigate the function of JAK2-STAT3 signaling pathway in pancreatic injury and systematic inflammatory response in rats with acute necrotizing pancreatitis ( ANP) . Methods SD rats were randomly divided into the ANP group (n=48), ANP+JAK2 inhibitor Ruxolitinib group (ANP+R group, n=48), ANP+STAT3 inhibitot Stattic group (ANP+S group, n=48), ANP+Ruxolitinib+Stattic group (ANP+R+S group, n=48), and sham operation group (SO group, n=48). 5％ sodium taurocholate injection via retrograde pancreatobiliary duct was used to establish ANP model. Blood samples from abdominal aorta and pancreatic tissue were collected after 3 h, 6 h, 12 h and 18 h after modeling. Serum amylase (AMY) and tumor necrosis factor-α(TNF-α) and interleukin-4 (IL-4) were tested. JAK2 and STAT3 mRNA expression and protein expression of p-JAK2 and p-STAT3 in pancreas were examined by RT qPCR and western blot, respectively. Results AMY, TNF-α and IL-4 in plasma, and JAK2 mRNA, STAT3 mRNA, p-JAK2 protein and p-STAT3 protein at different time points in ANP group were all obviously higher than those in SO group(P<0. 05). Serum AMY, TNF-αand IL-4 in ANP+R group, ANP+S group and ANP+R+S group at different time points were lower than those in ANP group [12 h (5391 ± 1009),(6130 ± 1227),(4818 ± 992)U/L vs (8524 ± 1360) U/L;(147.25 ± 27.85),(156.25 ± 23.17),(127.87 ± 21.39) ng/L vs (187.58 ±20.09)ng/L;(45.89 ±16.95),(50.19 ±15.87),(38.87 ±14.03)ng/L vs (58.85 ±9.34)ng/L] . JAK2 mRNA and p-JAK2 protein,STAT3 mRNA and p-STAT3 protein in ANP+R group and ANP+R+S group at different time points were obviously lower than those in ANP group (12 h 0. 357 ± 0. 091 vs 0. 597 ± 0. 121,1. 115 ± 0. 203 vs 1. 217 ± 0. 213,0. 361 ± 0. 089 vs 0. 489 ± 0. 097,0. 965 ± 0. 189 vs 1. 128 ± 0. 217, 0. 362 ± 0. 092 vs 0. 597 ± 0. 121,1. 107 ± 0. 212 vs 1. 217 ± 0. 213,0. 297 ± 0. 087 vs 0. 489 ± 0. 097,0. 713 ± 0. 184 vs 1. 128 ± 0. 217). STAT3 mRNA and p-STAT3 protein in ANP+S group were obviously lower than those in ANP group(0. 319 ± 0. 107 vs 0. 489 ± 0. 097,0. 849 ± 0. 177 vs 1. 128 ± 0. 217), and the difference was statistically different (P<0.05). Conclusions The activation of JAK2-STAT3 signaling pathway in pancreas may play a key role in the pathogenesis of systematic inflammatory response in ANP.

Objective:To evaluate the clinical efficacy of Buyang-Huanwu Decoction combined with modified Tingli-Dazao-Xiefei Decoction on the basis of conventional western medicine therapy in the treatment of chronic heart failure (CHF) with syndrome of qi deficiency and blood stasis. Methods:Seventy patients who met the inclusion criteria from November 2017 to November 2019 in Shijingshan District Hospital of Traditional Chinese Medicine were randomly divided into two groups, 35 in each group. The control group was treated with western medicine for chronic heart failure, and the treatment group was treated with Buyang-Huanwu Decoction combined with Tingli-Dazao-Xiefei Decoction on the basis of the control group. Both groups were treated for 2 weeks. The TCM syndrome scores were observed and compared before and after treatment. Minnesota Living with Heart Failure Questionnaire (MLHFQ) was used to evaluate the quality of life of patients. Lee’s Heart Failure Score was used to evaluate the severity of symptoms. The N-terminal pro-B type natriuretic peptide(NT-proBNP) was detected by ELISA. The adverse reactions during treatment were observed and the clinical efficacy was evaluated. Results:The total effective rate was 91.4% (32/35) in the treatment group and 77.1 % (27/35) in the control group, and the difference was statistically significant ( χ2=7.050, P=0.014). After treatment, the serum NT-proBNP in the treatment group [(1 725.3 ± 1 473.8) ng/L vs. (2 485.7 ± 2 164.4) ng/L; U=341.200, P=0.031] was significantly lower than that of the control group. The NT-proBNP [(54.3 ± 26.7) % vs. (35.5 ± 19.8)%; U=4.310, P=0.003] was significantly higher than that of the control group. After treatment, the TCM syndrome scores and MLHFQ scores in the treatment group were significantly lower than those in the control group ( t=3.785, 9.925, P=0.031, 0.001). During the treatment, no obvious adverse reactions were observed in both groups. Conclusion:On the basis of standardized treatment of Western medicine, Buyang-Huanwu Decoction and Tingli-Dazao-Xiefei Decoction can improve the clinical efficacy of CHF patients with qi deficiency and blood stasis syndrome, promote the repair of damaged myocardium (reduce NT-proBNP), and improve the quality of life of patients.