Objective To construct cDNA library of the treated Changliver cell by switching mechanism at 5′ end of RNA transcript (SMART) technique and analyze its quality.Methods cDNA of Changliver cell was aquired with reverse transcription polymerase chain reaction (RT-PCR) and long-distance PCR (LD-PCR),then the cDNA library was constructed with SMART cDNA library construction kit.Results Through testing,the high quality cDNA library containing full length cDNA of Changliver cell had been constructed.The titer of the amplified cDNA library was 4.5 × 1010 pfu*ml-1 and the average exogenous inserts of the recombinants was 1.5 kb.Conclusion These results suggest that the Changliver cell cDNA library has a high quality and lays a solid foundation for researching on Changliver cell and screening

Objective To treat femur fracture in neonates using a new homemade harness designed and observe the effec of this harness. Methods Designed a new harness used in femur fracture in neonates,and used this harness in 7(8 femur fracture)csses,including 6 boys(7 femur fracture)and 1 girl,the age from 1 day to 12 days(average 4.7 days).Those cases included proximal thirds(2 cases)and middle thirds(6 cases)of femur fracture.The angle of fracture was from 44°to 83°(average 62.4°)before treatment. Results The angle of fracture was from 0°to 22°(average 14.0°)after treatment using the harness.Hespitalization was from 2 to 3 days.There were no skin sloughing or harness breaking off.All cases were followed up 6 to 36 months(average 21.3 months).All femur fracture healed in good alignment with leg-length discrepancy＜1 cm.Movement of hip and knee was normal. Conclusions The harness is a better method to treat femur fracture of the proximal and middle thirds in neonates.Advantages of it include simply operation,minimal hospitalization,minimal cost,and easy nursing.

Objective To study the success rate of ventilator weaning in acute exacerbation of chronic obstructive pulmonary disease (AECOPD) patients influenced by bobbed endotracheal intubation. Methods One hundred and sixteen AECOPD patients who were given invasive mechanical ventilation and reached the standards of off-ventilator were divided into control group (general endotracheal intubation group) and experiment group bobbed (tracheal intubation) by random digits table method with 58 cases each. The respiratory rate (RR), heart rate (HR), rapid and shallow breathing index (RSBI), oxygenation index (OI), pulse blood oxygen saturation (SpO2), tidal volume and success rate of ventilator weaning were compared. Both groups were implemented of extubation if they reached the standards of the extubation time by observing their ability in spontaneous breathing test (SBT). Results The RR, HR and RSBI before extubation in experiment group were significantly lower than those in control group:(19.7 ± 2.3) times/min vs. (23.5 ± 2.3) times/min, (91.2 ± 6.3) times/min vs. (93.4 ± 8.1) times/min and 80.2 ± 6.7 vs. 90.5 ± 9.6, P<0.05, and the OI, SpO2, tidal volume and success rate of ventilator weaning were significantly higher than those in control group: (269 ± 9) mmHg (1 mmHg = 0.133 kPa) vs. (245 ± 16) mmHg, 0.929 ± 0.014 vs. 0.870 ± 0.037, (6.1 ± 1.2) ml/kg vs. (5.1 ± 0.8) ml/kg and 91.38%(53/58) vs. 77.59%(45/58), P<0.05. Conclusions The bobbed endotracheal intubations can improve the success rate of ventilator weaning in patients with AECOPD.

Objective To investigate the DNA methylation of skeletal muscle in spastic paralysis rats and correlation with the muscle fiber configuration. Methods 100 5-day old Wistar rats were randomly divided into model group and control group. The former was established the spastic paralysis modle and reared for 30 days. Then, tissues from the gastrocnemius of all the rats were observed with triplicate DNA methylation, myosin heavy chain-I (MHC-I) mRNA with RT-PCR and transmission electron microscopy. Results The DNA methylation was (4.95±0.83)×10% in the model group, significantly less than (6.59±0.75)×10% in the control group (P<0.001); while the MHC-I mRNA was (1.23±0.31), significantly more than (0.44±0.29) in the control group (P<0.001). The Z-line was disordered, and the mitochondria near the Z-line increased, with edema and partially broken in cristae. The balance between the thick and thin filaments was broken, and myofibrils envelope fused. Conclusion Hypomethylation and hyperexpression of MHC-I mRNA have been found in skeletal muscle of spastic paralysis rats, which may result in type I fibers increase. However, there was no sufficient evidence to support the correlation between the DNA methylation and the secondary pathological changes.

Objective To explore the optimal dose and ways of anesthesia for creating a rat model of spastic paralysis with intermittent bilateral common carotid artery ligation. Methods 60 Wistar rats were randomly divided into groups A, B, C, D, E and F. Group A was anaesthetized with 10% chloral hydrate 5 ml/kg injected subcutaneously, while group B with 4 ml/kg subcutaneously, group C with 4 ml/kg intraperitoneally, group D with 3 ml/kg subcutaneously, group E with 3 ml/kg intraperitoneally, group F with 2 ml/kg subcutaneously. The onset and duration of anesthesia, and intraoperative and postoperative mortality were compared. Results All the rats in the group A died during anesthesia, while the group F did not achieve the depth of anesthesia, and abandoned. The onset time was (6.5±0.7) min, maintained (121.4± 3.9) min, mortality was 0 in the group B, and it was (5.5±1.1) min, (122.0±3.6) min, 30% in the group C; (9.6±0.8) min, (106.7±3.7) min, 0 in the group D, (7.4±1.2) min, (105.3±3.5) min, 20% in the group E. The overall mortality rate was 0 in the groups accepted subcutaneous injected and 25% of intraperitoneal injection. Conclusion Anesthesia with 10% chloral hydrate 4 ml/kg subcutaneous injection is optimal of lower mortality, faster onset and longer maintaining in rats for spastic paralysis model with intermittent bilateral common carotid artery ligation.

Objective To evaluate the measurement uncertainty of flavonoids in loquat honey by UV spectrophotometry. Methods Through analyzing the whole process of the total flavonoids in loquat honey by UV spectrophotometry detection,the mathematical model was established,the uncertainty factors were determined,and each uncertainty was evaluated.Then the combined uncertainty was calculated and the expanded uncertainty of the measurement results at the 95％ confidence interval was given. Results The measurement uncertainty of the totle flavonoids content in loquat honey by UV spectrophotometry was (38.497±5. 674) μg?g-1 . Conclusion The uncertainty evaluation method is suitable for determination of the total flavonoids in loquat honey by UV method,and the uncertainty is mainly introduced by preparation of reference stock solution and standard curve fitting.

Objective To evaluate the clinical efficacy of arthroscopic treatment with anchor fixation for Segond fractures.Methods A total of 29 patients with Segond fractures treated with arthroscopic evaluation and anchor fixation technique between January 2011 and December 2015 were included in this retrospective case series study.There were 17 males and 12 females,with an average age of 41 years (range,27-62 years).Among the patients with Segond fractures,there were 20 patients combined with anterior cruciate ligament (ACL) rupture,three with posterior cruciate ligament (PCL) rupture,seven with medial collateral ligament (MCL) lesion,six with lateral collateral ligament (LCL) rupture,12 with meniscus lesion,and two with tibial plateau fracture.The time duration from injury to operation was 5-14 days (mean,7 days).The operation time and intraoperative blood loss volume were recorded.The postoperative pain,complications,knee joint stability,and fracture healing were observed.The pain was evaluated using numerical rating scale (NRS).Drawer test,Lachman test,lateral stress test,and Lysholm scores were performed to assess the knee function before and after operation.Results The operation time was 40-125 minutes (mean,85 minutes),and intraoperative blood loss was 10-30 ml (mean,15 ml).All patients were followed up for 12-18 months (mean,14 months).Two weeks after operation,the pain was not obvious,with NRS of 0-1.One patient had deep venous thrombosis one week after operation,and recovered after vascular intervention therapy.Other patients had no infection,deep vein thrombosis or other postoperative complications.The results of drawer test,Lachman test,and lateral stress test were negative in all patients,which indicated good knee joint stability.All patients were seen bone union,with the healing rate of 100％.The preoperative Lysholm scores were 35-62 points [(52.0 ± 4.2) points],and the scores one year after operation were 86-95 points [(91.9 ± 1.4) points] (P ＜ 0.05).Conclusion Arthroscopic treatment with anchor fixation has advantages of minor trauma,little hemorrhage,high fracture healing rate and good function recovery,and hence is an effective surgical treatment for Segond fractures.

Objective: To evaluate a self-designed external fixation restorer used for femoral shaft fractures in children. Methods: From September 2016 to October 2017, 19 children were treated at Department of Pediatric Orthopaedics, The Third Hospital of Hebei Medical University for irreducible femoral shaft fractures using our self-designed external fixation restorer. They were 15 males and 4 females, aged from 4 years and 2 months to 8 years (average, 6.3 years). There were 7 transverse fractures, 11 short oblique fractures and one oblique fracture. The restorer was applied directly to the femur for traction and temporary external fixation. The femoral shaft fractures were reduced closely before internal fixation with elastic stable intramedullary nails. The Flynn criteria for lower limb fracture were used to evaluate the curative effects postoperatively. Results: The operative time ranged from 32 to 45 minutes (37 minutes on average). All the fractures obtained closed reduction. No such intraoperative complications occurred as traction and compression injury to soft tissues like muscle, nerve and blood vessel. Follow-ups ranged from 8 to 20 months (average, 13 months). All the fractures got united after 7 to 15 weeks (average, 10.1 weeks). No implant failure or breakage occurred. The implants were removed 6 to 8 months after operation when the fractures got united. According to the Flynn evaluation criteria at the last follow-up, 18 cases were excellent and one was good. Conclusions Due to its advantages of simplicity, easy manipulation and direct action on the femur for traction, our self-designed external fixation restorer can improve the closed reduction for femoral shaft fractures in children so that its sustained and effective traction force and high quality of fracture closure avoid surgical opening. The temporarily fixation it provides after fracture reduction can facilitate intraoperative fluoroscopy of the femur.

Objective To compare the consistency between the estimated values of nine common resting energy expenditure estimation equations and the measured values among the elderly population in pension institutions. Methods A total of 181 elderly individuals from Henan Guangda Ouan Leling Nursing Center were recruited as study objects through cluster sampling. The differences between the estimated values of nine common equations and the measured values obtained by indirect calorimetry were compared, and subgroup analyses were conducted to compare their consistency across different subgroups. Results Among the nine equations, the Cunningham equation and the Owen equation showed relatively small deviations from the measured values, with no significant differences (deviation rates of -1.9% and -2.0%, root mean square errors of 222.7 kcal/d and 239.4 kcal/d, and coincidence rates of 48.4% and 47.5%, respectively). In subgroups stratified by gender and body mass index (BMI), the Cunningham equation showed no significant difference compared with the measured values. The Owen equation showed no significant difference compared with the measured values only in some subgroups [males with BMI of 20~ < 27 kg/m² and ≥27 kg/m², and females with BMI < 20 kg/m²]. Conclusion The Cunningham equation, which utilizes body composition data, can assess the resting energy expenditure of the elderly in pension institutions with relatively high accuracy and speed, demonstrating strong applicability in this population.

Objective:To investigate the effect of overexpression of T cell restricted intracellular antigen 1(TIA1)gene in M2-type tumor-associated macrophages(M2-TAMs)on invasion and migration of gastric cancer cells and its mechanism.Methods:Primary TAMs were extracted from gastric cancer tissues and induced to differentiate into M2-TAMs using IL-4 and IL-13.The TIA1 overex-pression plasmid(oe-TIA1)and its empty vector were transfected into M2-TAMs.The expression levels of TIA1 mRNA and protein were detected by qRT-PCR and Western blot.The expression levels of CD206 and CD163 were detected by flow cytometry.The levels of IL-10,TGF-β,VEGF-A and Arg-1 in cell culture supernatant were detected by ELISA.The transfected M2-TAMs were co-cultured with gastric cancer BGC-823 cells by Transwell co-cultivation system,and PI3K agonist 740Y-P was used to intervene in parallel.The cell migration ability was detected by scratch assay.Transwell assay was used to detect cell invasion ability.Protein expression levels of PI3K,p-PI3K,AKT,p-AKT,MMP-2 and MMP-9 in the cells were detected by Western blot.Results:①Compared with primary TAMs,the levels of CD206 and CD163 expression in M2-TAMs cells and IL-10,TGF-β,VEGF-A and Arg-1 in cell culture superna-tant were significantly increased(P<0.05),while the expression levels of TIA1 mRNA and protein were significantly decreased(P<0.05).Overexpression of TIA1 gene significantly decreased the expression levels of CD206 and CD163 in M2-TAMs and IL-10,TGF-β,VEGF-A and Arg-1 in cell culture supernatant(P<0.05).②Overexpression of TIA1 gene in M2-TAMs could significantly reduce the migration and invasion ability and the expression levels of p-PI3K/PI3K,p-AKT/AKT,MMP-2 and MMP-9 proteins in BGC-823 cells(P<0.05).③740Y-P could significantly reverse the inhibitory effects of overexpression of TIA1 gene in M2-TAMs on migration,inva-sion and PI3K/AKT signaling pathway of BGC-823 cells.Conclusion:Overexpression of TIA1 gene in M2-TAMs can affect the inva-sion and migration of gastric cancer cells by blocking the PI3K/AKT signaling pathway.