Objective To establish a rapid and accurate method of fluorescent quantitative PCR detection for Avian Influenza Virus H7N9 type.Methods A pair of primers and TaqMan fluorescent probes were designed specificly for H7N9 gene ac-cording to the published nucleotide sequence from National Center for Biotechnology Information.Positive recombinant plas-mid was built and the minimum copies of detection,repeatability and specificitv were tested.Results The results showed that this assay obtained positive recombinant plasmid,the linear relation was fine in the range from 102 to 108 copies/μl.The minimum copies was 500 copies/μl with this method,specificitv and repeatability(CV<1%)were fine,no cross reaction with other viruses.The precision was 100% (16/16)with this method for detecting 8 positive samples and 8 negative sam-ples.Conclution The establishment methods of fluorescent quantitative PCR detection could be used for the rapid diagnosis Avian Influenza virus H7N9 types.

Objective To study the effects of PNESF at different voltages on the life spans of ascites tumor bearing mice.Methods Forty ascites tumor bearing mice were divided into 5 groups and exposed to PNESF at 0,10,17,23 and 30 kV,respectively,for 30 minutes daily on 4 successive days.All of the animals then received routine care until death.Their eating habits,body weights and life spans were observed.A straight line fitting method was used for statistical analysis. Results The body weights of the mice exposed to 17 kV and 23 kV PNESF increased quickly during the four days of PNESF exposure.The weight gain in all groups declined after PNESF exposure was stopped.Thereafter the body weights of the mice exposed to the 17 kV PNESF increased more slowly than those in the control group.The mice exposed to the 23 kV PNESF gained weight more quickly than other groups.The mice exposed to the 17 kV PNESF lived the longest,demonstrating a lengthening of life of 25.5%compared with the control group.Conclusion Exposure to a PNESF of appropriate intensity might significantly increase the life span of ascites tumor bearing mice.

Objective: To investigate the role of X-box binding protein 1 (XBP1) in the treatment of human osteosarcoma HOS cells by pyropheophorbide-α methyl ester-mediated photodynamic therapy (MPPα-PDT), and to explore the possible mechanism. Methods: HOS cells were treated by MPPα-PDT for 6, 12 and 24 h, and the expression levels of inositol-requiring enzyme 1 alpha (IRE1α)-XBP1 pathway-related protein IRE1α and XBP1 were detected by Western blotting. The specific siRNA targeting XBP1 gene was transfected into HOS cells by lipofection, and treated by MPPα-PDT. The cells were divided into the blank group, siRNA-negative control (NC) group, siRNA-XBP1 group, MPPα-PDT group and MPPα-PDT+siRNA-XBP1 group. The expressions of XBP1 mRNA and protein were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The proliferation of HOS cells was detected by CCK-8 assay. The apoptotic rate was analyzed by flow cytometry (FCM). The expression levels of cleaved-caspase 3 and cleaved-poly ADP-ribose polymerase (cleaved-PARP) were determined by Western blotting. The intracellular level of reactive oxygen species (ROS) was detected by DCFH-DA probe staining, then the expression levels of Catalase and superoxide dismutase 1 (SOD1) proteins were detected by Western blotting. Results: The expression levels of IRE1α-XBP1 pathway-related proteins IRE1α and XBP1 were increased after the treatment of MPPα-PDT (both P < 0.05 ). siRNA-XBP1 inhibited the expression levels of XBP1 mRNA and protein (both P < 0.01). Silencing siRNA-XBP1 expression inhibited the proliferation activity of HOS cells (P < 0.01), up-regulated the apoptosis rate and the expression level of apoptosis-related protein cleaved-caspase 3 (both P < 0.05), and down-regulated the expression levels of antioxidant enzyme-related proteins Catalase and SOD1 (both P < 0.05). Compared with the MPPα-PDT group, the proliferation activity of HOS cells treated with MPPα-PDT+siRNA-XBP1 was decreased (P < 0.01), the apoptosis rate and the expression levels of cleaved-caspase 3 and cleaved-PARP were increased (all P < 0.05), the intracellular ROS level was up-regulated (P < 0.01), and the expression levels of Catalase and SOD1 were decreased (both P < 0.01). Conclusion: MPPα-PDT can induce the activation of IRE1α-XBP1 pathway in HOS cells. Silencing XBP1 can inhibit the proliferation activity of HOS cells, up-regulate the apoptosis rate, and increase the sensitivity of HOS cells to MPPα-PDT. The mechanism may be related to the up-regulation of intracellular ROS level and the down-regulation of anti-oxidation molecules.

Objective:To investigate the clinical efficacy of endoscopic magnetic compression bilio-enteric anastomosis for the treatment of biliary obstruction after complex abdominal surgery.Methods:The retrospective and descriptive study was conducted. The clinical data of 3 patients with biliary obstruction after complex abdominal surgery who were admitted to the First Affiliated Hospital of Xi′an Jiaotong University between January 2012 and December 2019 were collected. There were 2 males and 1 female, aged from 27 to 57 years, with a median age of 56 years. The 3 patients underwent endoscopic magnetic compression bilio-enteric anastomosis to complete internal drainage of bile ducts after several unsuccessful endoscopic or interventional therapy. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow up using outpatient examination was performed to detect the biliary stent, liver function and patency of anastomotic stoma up to December 2019.Results:(1) Surgical situations: all the 3 patients underwent successful endoscopic magnetic compression bilio-enteric anastomosis, including 2 cases with magnetic compression choledochoduodenal anastomosis and 1 case with magnetic compression choledochojejunostomy. The length of biliary stricture, length and width of magnetic device subsidiary magnet/patent magnet, time of magnetic compression for the 3 patients were 7.1 mm, 3.0 mm, 2.0 mm, 7 mm/8 mm, 6 mm/6 mm, 5 mm/5 mm, 130 minutes, 90 minutes, 75 minutes, respectively. (2) Postoperative situations: the time to biliary tract recanalization after operation for the 3 patients were 15 days, 8 days, 9 days, respectively. None of the patients encountered gastrointestinal perforation, hemorrhage or infection. (3) Follow-up: the biliary stents were inserted into the anastomotic stoma for the 3 patients after biliary tract recanalization, including a catheter of percutaneous transhepatic cholangiodrainage (PTCD) with 12 Fr size, a biliary plastic stent with 7 Fr size, a catheter of PTCD with 14 Fr size, respectively. The biliary stents were removed after 17 months, 2 months, and 6 months from the 3 patients, respectively. The 3 patients were followed up for 40 months, 20 months, and 5 months respectively after removing biliary stents, and the concentration of total bilirubin, concentration of aspartate aminotransferase, concentration of alanine aminotransferase for the 3 patients were 5-19 μmol/L, 25-40 U/L, and 20-35 U/L, respectively. The results of ultrasonic examination or magnetic resonance cholangiopancreatography examination showed that no intra-hepatic bile duct dilation or stricture of choledochojejunostomy was found within the 3 patients. One of the 3 patients was hospitalized for biliary tract infection after 37 months from removing biliary stents, and the results of ultrasonic examination or magnetic resonance cholangiopancreatography examination showed intrahepatic cholelithiasis. The cholelithiasis was removed under endoscopy and stricture of choledochojejunostomy was not found within patient by the endoscopic examination or cholangiography examination.Conclusion:The endoscopic magnetic compression bilio-enteric anastomosis is a safe and feasible technique for the treatment of biliary obstruction after complex abdominal surgery with good long-term effects.

Objective:To characterize limb traumatic osteomyelitis in a regional trauma center in South China.Methods:The case system at Division of Orthopaedics and Traumatology, Department of Orthopaedics, Nanfang Hospital was searched for the data of confirmed limb traumatic osteomyelitis from January 1, 2010 through September 1, 2019. The clinical items collected were gender, age of onset, cause of injury, injury nature (open or closed), infected site (single site or multiple sites), intraoperative culture of pathogenic microorganisms, number and type of pathogenic microorganisms, and amputation. The above data were sorted out and analyzed statistically.Results:A total of 674 patients suffered limb traumatic osteomyelitis in South China with a male to female ratio of 4.81∶ 1. The limb traumatic osteomyelitis resulted mainly from open injury (64.09%, 432/674). It was mainly caused by a traffic accident (39.51%, 211/534). It was featured by single-site infection (83.68%, 564/674). It affected mainly the lower limbs: the tibia (53.19%, 300/564), the femur (18.97%, 107/564), the calcaneus (11.70%, 66/564) and other foot bones (4.61%, 26/564). The positive rate of intraoperative microorganisms culture was 65.26% (355/544). In the majority of patients (74.65%, 265/355), the traumatic osteomyelitis was caused by infection of a single pathogenic microorganism. The most common single pathogenic microorganism was Staphylococcus aureus (38.11%, 101/265), followed by Pseudomonas aeruginosa (20.00%, 53/265). The rate of amputation related to traumatic osteomyelitis was 3.71% (25/674), and the overall disability rate was 4.45% (30/674).Conclusions:In South China, more males were prone to limb traumatic osteomyelitis. Most cases were secondary to an open fracture and caused by a traffic accident. Traumatic osteomyelitis usually occurred at a lower limb. The most common pathogenic microorganism was Staphylococcus aureus.

Objective:To investigate the protective role of extracellular signal-regulated kinase (ERK) signaling pathway in the process that vasonatrin peptide (VNP) reduces hepatic ischemia-reperfusion injury in rats.Methods:Twenty SD rats, weighting 200-250 g, were randomly divided into four groups and each group has five rats. The four groups were sham operation group (S group), ischemia-reperfusion group (I/R group), VNP group (V group) and PD98059+ VNP group (P+ V group). In the rat model of hepatic warm ischemia and reperfusion, the hepatic artery and portal vein of the left lobe and middle lobe of the liver were clamped with arterial clamp for 45 min followed by reperfusion for 120 min. In the V group, VNP (50 μg/kg) was injected 10 minutes before ischemia. In the P+ V group, PD98059 (2 mg/kg) was injected 20 min before VNP injection followed by VNP administration and I/R treatment. The serum levels of alanine amino transaminase (ALT), aspartate amino transferase (AST), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and the superoxide dismutase (SOD) in liver tissue homogenate and malondialdehyde (MDA) were measured. The histopathology of liver tissue was observed. The contents of p-ERK1/2 were detected by Western blot.Results:Compared with S group, in I/R group and P+ V group the serum levels of ALT [(489.65±11.22), (333.05±24.77) vs. (33.78±4.88) U/L], AST [(651.43±14.99), (503.18±21.48) vs. (154.84±12.32) U/L], TNF-α [(12.83±1.09), (9.64±0.57) vs. (2.11±0.11) ng/L], IL-1β [(7.19±0.62), (5.12±0.22) vs. (1.10±0.49) ng/L], MDA [(8.00±0.88), (5.60±1.01) vs. (2.76±1.29) μmol/mg] increased, while SOD [(54.89±10.60), (68.85±8.33) vs. (126.10±15.63) nmol/mg]decreased (all P<0.05). The histopathology of liver tissue revealed that liver structure damaged more seriously in I/R group and P+ V group. Western blot analysis showed that p-ERK1/2 decreased significantly in I/R group and P+ V group. Compared with I/R group, ALT, AST, MDA, TNF-α and IL-1β decreased significantly and SOD increased significantly in V group (all P<0.05). The histopathology of liver tissue revealed that liver structure was damaged slightly, and p-ERK1/2 increased significantly in V group compared with I/R group ( P<0.05). Conclusion:VNP can significantly reduce hepatic ischemia-reperfusion injury through activation of p-ERK1/2 signaling pathway and inhibition of hepatocyte inflammatory response.

Objective: To investigate the health status of occupational mercury workers and reveal the effects of mercury exposure on the cardiovascular system. Methods: In June 2019, a total of 2651 mercury workers participated in the occupational health examination between 2016-2018 from a thermometer manufacturing plant and a fluorescent lamp manufacturing plant were included in this study. Then, they were divided into a high-level mercury exposure group (425 workers whose urine mercury concentration >35 μg/g creatinine) and a low-mercury mercury exposure group (2226 workers whose urinary mercury concentration <35 μg/g creatinine) . Mercury concentration in the workplace was also detected. Finally, the results of electrocardiogram (ECG) , blood routine, blood biochemistry and other physical examinations were analyzed. The measurement data of age and exposure years were analyzed by test. Urinary mercury and blood parameters were analyzed by Mann-Whitney nonparametric test. Chi-square test was used for the analyses of gender, ECG abnormality rate and other categorical data. Results: The 8-hour weighted average allowable concentration (CTWA) of mercury in the workplace of high-exposure group was 0.002 2-0.152 mg/m³. The abnormal rate of ECG in the high-exposed group (29.6%) was higher than that in the low-exposure group (10.1%) in 2018 (P<0.01) . Compared with the low-exposure group, the WBC of the high-exposure group from 2016 to 2018 was increased, with statistically significance (P<0.05) ; the RBC of the high-exposure group in 2016 and 2017 was decreased, with statistically significance (P<0.01) ; the total bilirubin concentration in the high-exposure group was decreased from 2016 to 2018, with statistically significance (P<0.05) . Conclusion Long-term exposure to high concentration of mercury in the workplace may influence cardiovascular system. Therefore, engineering protection and individual protection should be implemented well.

A biliary contrast agents pushing device, including a syringe pushing system and a remote controller is introduced. The syringe pushing system comprises an injector card slot, a support platform and an injection bolus fader. A 20 mL syringe can be fitted on the syringe pushing system and kept with the ground about 30 degree. This system can perform air bubble pumping back and contrast agents bolus injection as well as speed adjustment. Remote controller is an infrared remote control which can start and stop the syringe pushing system. With this device, the remote controlled cholangiography technology can be achieved, which can not only protect doctors from X-ray radiation but also improve the traditional T-tube cholangiography and the contrast effect, reduce postoperative complications in patients as well. The application of this device will improve the current diagnosis and treatment system, the device will benefit the majority of doctors and patients.
Contrast Media ; administration & dosage ; Humans ; Injections ; Postoperative Complications ; Syringes

A motion unit for sucking robot with a stable motion, convenient operation and process simulation is introduced. The key parameters and process data of the sucking operation were obtained from the clinical work, which provided the basis for the design of the sucking robot motion unit. According to the points of sucking action, robotic thumb, forefinger and metacarpophalangeal joints were used to grip the suction tube, and the servo and arm structure were used to simulate the motion of the wrist and elbow to complete the rotation and push of the sputum suction tube. The feasibility is verified through the advanced sputum suction training model. The movement unit is stable in movement, and can smoothly complete the clamping, feeding, back off protection and rotating tube removal of the sputum suction tube, so as to achieve effective sputum suction.
Catheterization ; Intubation, Intratracheal ; Robotics ; Suction

BACKGROUND: Lung cancer is the most common malignancy world-wide. There are a variety of immune infiltrating cells in tumor microenvironment, which is an important component of tumor immunity and has clinical significance for the prognosis of patients. CD45RO is a surface marker of memory T cells. The expression of CD45RO⁺ tumor infiltrating lymphocytes (TILs) is associated with the prognosis of many tumors. The purpose of this study was to evaluate the relationship between the density of CD45RO⁺ TILs in tumor and stromal area and the clinical characteristics of patients with non-small cell lung cancer (NSCLC) and its impact on the prognosis of patients. We aimed to explore the clinical value of CD45RO⁺ TILs and programmed cell death ligand 1 (PD-L1) as prognostic markers. METHODS: Multiple fluorescent immunohistochemical staining was used to stain the tissue microarray chips of 167 patients with NSCLC, marking CD45RO, cytokeratin (CK) and PD-L1. Using artificial intelligence image recognition technology and tumor cell-specific CK staining, divide the tumor and stromal area in the tissue, evaluate the density of CD45RO⁺ TILs in the tumor and stromal area, and the expression level of PD-L1 in tumor cells. The non-parametric test was used to analyze the relationship between CD45RO⁺ TILs and the clinical characteristics of patients, and the Kaplan-Meier method and Cox risk ratio model were used to analyze the relationship between CD45RO⁺ TILs independently or in combination with PD-L1 and tumor prognosis. RESULTS: The density of CD45RO⁺ TILs was significantly associated with patient age, smoking, tumor stage, and pathological type. Single-factor survival analysis showed that NSCLC (P=0.007) stromal region and lung adenocarcinoma (LUAD) (P<0.001) with CD45RO⁺ TILs high density had better OS. Multivariate survival analysis showed that the high density of CD45RO⁺ TILs in the stromal region of NSCLC (HR=0.559, 95%CI: 0.377-0.829, P=0.004) and lung adenocarcinoma (HR=0.352, 95%CI: 0.193-0.641, P=0.001) were independent prognostic factors for overall survival time (OS). Combined with PD-L1 score of tumor cells in tumor tissues and infiltration score of CD45RO⁺ TILs in all tumor tissues, the patients were divided into 4 groups: patients with PD-L1⁺/CD45RO⁺ had the longest disease-free survival (DFS) time, and patients with PD-L1⁺/CD45RO- had the shortest DFS time. Multivariate Cox regression analysis showed that PD-L1⁺/CD45RO- was an independent prognostic factor for DFS and had a higher risk of poor prognosis compared to the other three groups (HR=2.221, 95%CI: 1.258-3.919, P=0.006). CONCLUSIONS In tumor tissues, the density of CD45RO⁺ TILs, as well as the combination of CD45RO⁺ TILs and PD-L1 in tumor areas, significantly correlated with clinicopathological features and prognosis of NSCLC, which can be used as a new prognosis marker.