Objective Tyrphostin AG1024(3-Bromo-5-t-butyl-4-hydroxybenzylidenemalonitrile) is a specific insulin like growth factor type Ⅰ receptor tyrosine kinase blocker,this study is to investigate the effect of AG1024 on the proliferation and apoptosis of hepatocellular carcinoma cell lines.Methods Treated with AG1024 on vailed concentrations(0～40 μmol/L),human hepatocellular carcinoma cel lines HepG2 and SMMC-7721 were observed for morphological and molecular biology changes,the effect of AG1024 on the cell lines proliferation invasion ability as well as apoptosis was evaluated. Results MTT showed that AG1024 dose-dependently inhibited the proliferation of hepatocellular carcinoma cells,flow cytometry suggested that AG1024 significantly promoted cell lines apoptosis,the cell invasion assay indieated that AG1024 significantly inhibited cell's invasion ability.RT-PCR showed over-expression of IGF-IR in liver cancer cells.and AG1024 dose-dependently increasedtheexpressionofcytochreme C. According totheresultsof Western, blotting,the phosphor-ERK and procaspase-3 were down-regulated while the total ERK remained unchanged. Conclusion AG1024 as a specific IGF-IR blocker blocks the downstream signaling cascade and thus inhibits the proliferation of hepatocellular carcinoma cells and induces cell's apoptosis.

Background and purpose:Visceral pleural invasion (VPI) and vessel invasion (VI) are poor prognostic factors in patients with non-small cell lung cancer (NSCLC). The primary initial recurrence site may be local recurrence in VPI and distant metastasis in VI. The purpose of this study was to validate the prognostic impact and effect of the initial recurrence site of VPI and VI on survival outcomes for NSCLC.Methods:Two hundred and ninety patients who were diagnosed as having NSCLC and underwent lobectomy between Jan. 2007 and Dec. 2013 were retrospectively analyzed. VPI was identiifed in 51 patients as VPI group, the other 239 patients without VPI as non-VPI group. VI was identiifed in 29 patients as VI group, the other 261 patients without VI as non-VI group. Clinical characteristics, overall survival (OS), disease-free survival (DFS) were compared.Results:There were statistically signiifcant differences between VPI group and non-VPI group in tumor size, lymph node metastasis, TNM stage and initial recurrence site (P<0.05). Furthermore, there were statistically signiifcant differences between VI group and non-VI group in lymph node metastasis and TNM stage (P<0.05). The 1-, 3- and 5-year OS rates in VPI group (88.2%, 56.7% and 52.7%) were lower than those in non-VPI group (95.8%, 83.7% and 74.0%,P<0.001). The 1-, 3- and 5-year OS rates in VI group (79.3%, 56.8% and 48.7%) were lower than those in non-VI group (96.1%, 81.3% and
72.3%,P=0.001). Cox regression showed TNM stage was a significant prognostic factor for DFS, whereas lymph node metastasis and VPI were signiifcant prognostic factors in patients with NSCLC.Conclusion:The primary initial recurrence site in VPI patients is local recurrence. Patients with VPI or VI may need more postoperative therapy because of their poor prognosis.

Aim To discuss the repairing mechanism of Qinbai Qingfei Concentrated pellets to lung tissue of rats infected by mycoplasma. Method 60 Wistar rats weighting 80~100 g, male to female:1 ∶ 1) were di-vided into six groups randomly ( 10 rats in each group): blank group, model group, positive group, the high、middle and low dose groups of Qinbai Qingfei Concentrated pellets. Rats were infected through nasal intubation drip of MP. After 10 days of administration, concentrations of IL-6 , IL-8 AND TNF-α in serum of MPP rats were detected. Left pulmonary tissues of rats were collected to observe the lung tissue pathological change by HE staining and right pulmonary tissues were used to detect the transforming growth factor-beta ( TGF-β) and surface activity related protein A( SP-A) mRNA expression level by real-time quantitative PCR ( RT-PCR) and TGF-βand SP-A protein expression by (Western blot. Result Qinbai Qingfei Concentrated) pellets significantly inhibited inflammation of lung tis-sue, reduced the expression of TGF-β and increased the expression of SP-A in the lung tissue of rats infec-ted by mycoplasma. Conclusion Qinbai Qingfei Con-centrated pellets can inhibit epithelial-mesenchymal transition ( EMT ) , of alveolar type Ⅱ epithelial cells by reducing the content of TGF-β and restore the nor-mal morphology and function of the lung by increasing the expression of SP-A.

Objective To investigate the effect of gemcitabine, paclitaxel combined with nedaplatin in treatment of locally advanced nasopharyngeal carcinoma. Methods 40 patients diagnosed as locally advanced nasopharyngeal carcinoma from May 2012 to August 2014 were randomly divided into observation group and control group. Patients in the observation group received gemcitabine plus nedaplatin chemotherapy, while those in the control group received paclitaxel plus nedaplatin chemotherapy. Then chemotherapy effect, adverse reactions, survival time and tumor marker contents were compared. Results The effective rate of the observation group was 65 % (13/20), which was higher than that (30 %, 6/20) of the control group (χ2 = 4.912, P< 0.05), however, there were no statistically differences in gastrointestinal reactions, bone marrow suppression, liver damage, kidney damage between groups (P> 0.05). Before chemotherapy, the SCCAg and CYFRA21-1 content of the control group were (2.30 ±0.31) ng/L and (18.27±2.19)μg/L, and the observation group were (2.34±0.27) ng/L and (18.48±2.25)μg/L, and there was no significant difference between the groups (P>0.05). After chemotherapy, the SCCAg and CYFRA21-1 content of the control group were (1.92±0.22) ng/L and (13.72±1.74) μg/L, and the observation group were (1.20 ±0.15) ng/L and (8.49 ±0.91) μg/L (P= 0.000). Conclusion Gemcitabine combined with nedaplatin chemotherapy can improve the chemotherapy effect and reduce tumor marker content, without increasing more adverse reactions, which is an ideal chemotherapy regimen for advanced nasopharyngeal carcinoma.

Objective:To express and purify hemoglobin receptor(HgbA) and its partial fragment(HgbAF) from Haemophilus ducreyi and to develop a sandwich ELISA for the detection of H.ducreyi infection.Methods:The HgbA,a hemoglobin-binding outer membrane protein of H.ducreyi and its partial fragment(HgbAF) were expressed by cloning the genes of hgbA and its 705bp fragment into pET30a and pET28a respectively,and the expressing products were purified from E.coli BL21 with Ni-NTA-His affinity chromatography.The polyclonal antibodies were developed by immunizing rabbits with the rHgbA and rHgbAF.The anti-rHgbA IgG and anti-rHgbAF IgG were purified respectively by saturated amine sulphate precipitation,and their immunoactivity with rHgbA and rHgbAF was tested by Westen blot and ELISA analysis.A Sandwich ELISA was developed for the detection of chinical infection of H.ducreyi using the specific polyclonal antibodies.Results:The HgbA and its partial fragment,HgbAF of H.ducreyi,were expressed and purified successfully by cloning their genes respectively.The results obtained by Western blot analysis showed that each of the antibodies could react with both antigens,rHgbA and rHgbAF.The results of the ELISA analysis showed that H.ducreyi strain was strongly positive,and all other bacteria,including H.influenzae and the bacteria known to relate to genital ulcers were negative.The results of the ELISA analysis showed that the minimum amount of rHgbA detected was 1.56 ng/ml and the minimum number of CFU of H.ducreyi detected was 2?105 cfu/ml in buffer and 1?106 cfu/ml in pus.Conclusion:HgbA and its partial fragment,HgbAF of H.ducreyi are expressed and purified successfully.The polyclonal antibodies developed by immunizing rabbits using rHgbA and rHgbAF could react not only with rHgbA and rHgbAF,but also with H.ducreyi specifically.They do not react with other bacteria,including H.influenzae and the bacteria known to relate to genital ulcers.So the ELISA based on the polyclonal antibodies was specific for the detection of H.ducreyi.Although the level of sensitivity of the ELISA may not be sufficient to detect H.ducreyi in all clinical specimens,further work to increase the sensitivity could potentially make this assay a valuable tool in areas where chancroid is endemic.

Objective To provide an objective basis for differential diagnosis of pancreatic diseases through quantitative analysis of the different features of contrast-enhanced endoscopic ultrasonography (CE-EUS). Methods A total of 32 patients with suspected or confirmed pancreatic neoplasms or chronic pancreatitis and 19 patients who underwent EUS due to other digestive problems other than pancreatic disease were enrolled. Features of blood perfusion of the regions of interest during CE-EUS were analyzed quantitatively. The findings were compared with cytological and/or histopathological results of EUS-FNA and/or surgery.Results Quantitative analysis of CE-EUS showed peak intensity (PI) value of 19 normal pancreas was 0.648 ±0. 174, which was statistically different from that of pancreatic cancer and pancreatic cystic lesions. Based on ROC, the cutoff of differential diagnosis was 0. 505, and the sensitivity and specificity were 100. 0% and 84. 2%, respectively. PI value of 6 chronic pancreatitis was the highest (0. 772 ±0. 106). In pancreatic neoplams, PI values of pancreatic carcinoma, pancreatic cyst and pancreatic endocrine tumors were significantly different. Based on a cutoff of 0. 195, the sensitivity and specificity of differentiation of pancreatic carcinoma and pancreatic cyst were 85.7% and 87.5%, respectively. PI value of 14 pancreatic carcinoma and that of 4 pancreatic endocrine tumors were 0. 321 ± 0. 119 and 0. 763 ± 0. 115, respectively. Through the comparison between the AT and TTP of the focal lesions and surrounding pancreatic parenchyma, 78.6% pancreatic carcinoma showed slow falling-in and rapid wash-out and all the endocrine tumors showed rapid falling-in and rapid wash-out. The PI value of 8 patients with pancreatic cyst was 0. 181 ±0. 036, with no enhanced blood flow in the cyst. The TIC was a straight line. Conclusion CE-EUS with quantitative analysis is a promising method that can be a more objective basis in the differential diagnosis of pancreatic diseases.

Objective:To evaluate the efficacy of individualized preoperative planning combined with various fixations in the treatment of complex multiplanar deformities at lower extremity.Methods:Four patients with complex multiplanar deformities at lower extremity were treated at Department of Orthopedics, The First Affiliated Hospital to Harbin Medical University from January 2018 to May 2019. They were one male and 3 females, aged from 12 to 65 years (average, 28.8 years). All patients underwent thin-layer (1 mm) CT scan before surgery. Individualized preoperative planning and surgical simulation were conducted using the CT data. 3D printing technology was used to fabricate individualized surgical templates to facilitate osteotomy. Recorded were operation time, frequency of intraoperative fluoroscopy, fracture healing time, Lysholm scores before and after operation, Hospital for Special Surgery (HSS) knee function scores, pain scores of visual analogue scale (VAS) and postoperative complications in this cohort.Results:The operation time averaged 160.0 min, the frequency of intraoperative fluoroscopy 5.0 times, the follow-up duration for the 4 patients 8.8 months, the fracture healing time 6.3 months, the Lysholm scores before and after operation 55.8 and 80.5 points respectively, the HSS scores before and after operation 61.0 and 80.8 points respectively, and the VAS scores before and after operation 3.8 and 1.0 points respectively. Accurate osteotomy was achieved with assistance of the templates and no deformation or breakage of the templates was observed. Follow-ups showed fine alignment of the lower extremities by the full-length X-ray films and no recurrence of deformity, infection, implant breakage, or delayed union or non-union of the osteotomy sites.Conclusions:For patients with multiplanar deformities at lower extremity, individualized preoperative planning can be made before surgery to obtain the accurate parameters relevant to the osteotomy. In combination with various fixations, a patient-specific template made by 3D printing can be used to achieve accurate osteotomy, ensuring good efficacy, safety, efficiency and accuracy of the surgery.

Objective To rapidly establish a mouse model for optical imaging of the dynamical process of pseudotyped Middle East respiratory syndrome coronavirus (MERS-CoV) infection. Methods In vitro expression in HeLa cells and functions of hDPP4 encoded by recombinant adenovirus Ad5 and pseudo-typed MERS-CoV were verified. The recombinant adenovirus expressing hDPP4 (Ad5-hDPP4) was injected into BALB/ c mice, which were then injected with pseudotyped MERS-CoV expressing firefly luciferase at a titer of 3×107 TCID50(50％ tissue culture infective dose) via intrathoracic (I. T. ) or intraperitoneal (I. P. ) injection. MERS-CoV infection and tissue distribution were observed using optical imaging techniques. Re-sults hDPP4 and firefly luciferase were efficiently expressed in HeLa cells. In BALB/ c mice injected with Ad5-hDPP4 via I. P. , firefly luciferase expression were detected in abdomen between 48-96 h after pseudo-typed MERS-CoV infection. The expression of firely luciferase was also detected in chests of BALB/ c mice injected with Ad5-hDPP4 via I. T. around 48 h after pseudotyped MERS-CoV infection. Conclusions This study reported a simple and rapid method for establishing a mouse model for conveniently and dynamically monitoring pseudotyped MERS-CoV infection, which might provide an effective means for in vivo evaluation of neutralizing antibodies or entry inhibitors by visualization with optical imaging techniques.

Objective:To observe the effect of taping on the kinematic characteristics of the ankle joint during forward and lateral jumps by male basketball players with chronic ankle instability (CAI).Methods:A Vicon 3D motion capture system and a Kistler 3D ergometer were used to collect data describing the landing data with or without taping from forward and lateral jumps of 29 male basketball players with CAI. The landing data included the dorsiflexion and plantarflexion angles, valgus and inversion angles and external and internal rotation angles. Dorsiflexion or plantarflexion angular velocity was also recorded along with valgus or inversion angular velocity and external or internal rotation angular velocity 200ms, 150ms, 100ms and 50ms before and after touchdown. The data obtained were modeled using three-dimensional motion analysis software, and then analyzed.Results:Taping reduced the ankle plantarflexion in landing from a forward jump by 3.27° 50ms before landing and by 2.70° at touchdown. The ankle inversion angle was reduced 2.13° 50ms before touchdown, while the angle of external rotation decreased by 2.59° 200ms before touchdown and 2.17° 150ms before. Moreover, the angle of external rotation 100ms after landing was reduced by a significant 1.59° compared with that without taping. In lateral jumps taping reduced the average ankle plantarflexion angle by 1.94° 50ms before landing and 3.23° at touchdown compared with no taping. Ankle inversion was reduced significantly by 2.86° 50ms before landing and by 2.87° at touchdown. External rotation was a significant 0.93° less 200ms before landing and 2.36° smaller 150ms before touchdown. In the forward jump landing, taping reduced the average angular velocity of ankle dorsiflexion on landing by a significant 58.5°/s and by 28.39°/s 100ms later. In the lateral jump landings the average ankle dorsiflexion velocity decreased by significant 20.5°/s with taping, but the valgus velocity increased by 49.7°/s compared with no taping. However, 50ms after touchdown the speed of external rotation with taping was 30.3°/s slower than without taping.Conclusions:Ankle taping can modify ankle rotation angles and angular velocities during landing from jumps. This is particularly helpful for basketball players with CAI.

Objective:To investigate the changes in gut microbiota diversity with age in elderly people using high-throughput sequencing.Methods:Ninety healthy volunteers were recruited. People who were <60 years old (middle-aged group) were set up as a baseline control group (Age A group), while those aged ≥60 years old were further divided into four groups (60-<70: Age B group, 70-<80: Age C group, 80-<90: Age D group, ≥90: Age E group). Fecal samples were collected to extract DNA. The second-generation sequencing technology was used to amplify and sequence the V3-V4 hypervariable region of 16S rDNA. Bioinformatics analysis was performed to compare the differences in gut microbiota and functional genes among groups.Results:At the phylum level, gut microbiota were composed mainly of Firmicute, Bacteroidetes, Proteobacteria and Actinobacteria in different groups. The proportion of Firmicute was the highest, accounting for over 60%, followed by that of Bacteroidetes. At the genus level, the abundance of Faecalibacterium genus decreased with age. The α diversity analysis showed that the gut microbiota in the elderly of different ages had higher abundance and uniformity, and there was no significant difference among groups. However, the β diversity analysis showed that in community structure there was difference between Age A and Age B groups, and similarity between Age B and Age C groups. Conclusions:The community structure of gut microbiota changed significantly between young and middle-aged people and the elderly over 60 years old. It tended to be relatively stable in people of 60-80 years old, but changed again when they were over 80 years old. Chronic inflammatory diseases, metabolic diseases and tumors in the elderly might be associated with the decrease in Faecalibacterium.