Purpose: Mixed-lineage leukemia protein 4 (MLL4/KMT2D) is a histone methyltransferase, and its mutation has been reported to be associated with a poor prognosis in many cancers, including lung cancer. We investigated the function of MLL4 in lung carcinogenesis. Materials and Methods: RNA sequencing (RNA-seq) in A549 cells transfected with control siRNA or MLL4 siRNA was performed. Also, we used EdU incorporation assay, colony formation assays, growth curve analysis, transwell invasion assays, immunohistochemical staining, and in vivo bioluminescence assay to investigate the function of MLL4 in lung carcinogenesis. Results: We found that MLL4 expression was downregulated in non–small cell lung cancer (NSCLC) tissues compared to adjacent normal tissues and tended to decrease with disease stage progression. We analyzed the transcriptomes in control and MLL4- deficient cells using high-throughput RNA deep sequencing (RNA-seq) and identified a cohort of target genes, such as SOX2, ATF1, FOXP4, PIK3IP1, SIRT4, TENT5B, and LFNG, some of which are related to proliferation and metastasis. Our results showed that low expression of MLL4 promotes NSCLC cell proliferation and metastasis and is required for the maintenance of NSCLC stem cell properties. Conclusion Our findings identify an important role of MLL4 in lung carcinogenesis through transcriptional regulation of PIK3IP1, affecting the PI3K/AKT/SOX2 axis, and suggest that MLL4 could be a potential prognostic indicator and target for NSCLC therapy.

Objective    To investigate the feasibility of magnamosis rings designed based on magnetic compression technique in esophageal anastomosis reconstruction. Methods    According to the anatomical characteristics of esophagus in SD rats, the esophageal magnamosis rings were designed. SD rats were used as animal models (n=10, 5 males and 5 females) to complete the magnetic anastomosis reconstruction of the cervical esophagus using magnamosis rings, and the operation time, animal survival, postoperative complications, magnetic rings excretion time were recorded. Two weeks after operation, the rats were killed, and the esophageal anastomotic specimens were obtained. The blasting pressure of the anastomotic site was measured and the formation of the anastomotic site was observed with naked eyes. Results    Esophageal magnamosis was successfully performed in 10 SD rats, and the median operation time was 11 (8-13) min. All rats survived without anastomotic leakage, anastomotic stenosis, or magnetic rings incarceration. The magnetic rings were discharged after 8 (5-10) days and the burst pressure was higher than 300 mm Hg. Visual observation showed that the anastomotic muscle healed well and the mucosa was smooth. Conclusion    The magnetic compression technique  can be used for anastomosis reconstruction of esophagus, which has the advantages of simple operation and reliable anastomosis effect, and has clinical application prospect.

Objective: To investigate the effect and the mechanism of Astragalus membranaceus (Huangqi in Chinese, HQ) extract on the intestinal absorption of six alkaloids of Aconitum carmichaelii (Fuzi in Chinese, FZ) in rats with spleen deficiency and provide novel insights into the application of HQ on modulating intestinal barrier. Methods: Four-week-old male Sprague-Dawley rats were fed with Xiaochengqi Decoction to induce the spleen deficiency model for 40 d. Single-pass intestinal perfusion model were used to study the effects of HQ extract on the absorption of alkaloids. Protein expression and mRNA levels of MRP2 and BCRP and tight junction proteins (TJ, including Claudin-1, Occludin and ZO-1) were measured using Western blot and real-time PCR, respectively. The location and expression of TJ protein was also investigated by the immunofluorescence method. Results: Compared with the normal group, the protein expression of MRP2, BCRP and TJ proteins in the model group were significantly down-regulated. After oral administration of HQ, the alkaloid absorption in intestinal villi was inhibited, MRP2, BCRP and TJ proteins were up-regulated, the green fluorescence staining of Claudin-1, Occludin, and ZO-1 was enhanced, and a thick layer of mucus was deposited on the surface of the epithelium of the intestinal cavity. Conclusion: HQ as an intestinal barrier modulator improves the physiological changes of the intestinal environment of spleen deficiency to reduce the absorption of toxic components, leading to a decrease in the absorption of drug-like molecules.

Objective: This study aimed to explore the role of SGK-1 in the occurrence of temporomandibular joint(TMJ) osteoarthritis(TMJ-OA). Methods: Sixteen rats were randomly divided into Control group(Control group) and TMJ-OA group(TMJ-OA group), and TMJ-OA group was injected with sodium iodoacetate(MIA) intra articular cavity while Control group was injected with 0.9% sodium chloride solution. Pain behavior was assessed by measuring the head withdrawal threshold(HWT) with a von-Frey apparatus. Hematoxylin-eosin(HE) and Safranin O-fast green stains were used to observe the histological structure changes of the condyle of TMJ-OA rats. Real-time PCR was performed to exam the expression levels of mRNA of SGK-1, MMP-13, IL-1β, COX-2 in mandibular condylar cartilage(MCC). HE stain was used to observe the histological structure changes of the trigeminal ganglion(TG) of TMJ-OA rats. Real-time PCR was performed to exam the expression levels of mRNA of SGK-1, COX-2 in TG. Results: MIA injection induced typical OA-like lesions in the TMJ within 28 days. Administration of MIA led to the significant decrease in HWT, disordered of the condyle cartilage and subchondral bone structure, demyelination aggravated of nerve fibers in TMJ rats. Compared with rats in Control groups, the expression levels of SGK-1 in MCC and TG of rats in TMJ-OA group were upregulated. Conclusion In the pathological process of TMJ-OA, SGK-1 may plays an important role not only in cartilage structural damage but also in pain transmission.

Based on the principle of magnetic anastomosis technique, the design of magnetic anastomosis system for endoscopic tissue clamping is proposed. The system includes a semi-ring magnet, a special structure transparent cap and a detachable push rod. With the help of the existing digestive endoscopy and endoscopic tissue gripper, the endoscopic close clamping and anastomosis of the bleeding or perforated tissue can be completed. After the anastomosis, the magnet falls off and is discharged through the digestive tract. Animal experiments showed that the system was easy to use, the fistula was clamped firmly, the magnet was discharged for 7~21 days, and there was no magnet retention and digestive tract obstruction. Further safety verification, optimization of endoscopic operation, the system can be used in clinical trial.
Anastomosis, Surgical ; Animals ; Constriction ; Endoscopy, Gastrointestinal ; Magnetics ; Magnets

Objective To systematically review the clinical efficacy and safety of early use of recombinant human erythropoietin (rHu-EPO) for neuroprotection in premature infants.Method From establishment of the databases to November 2017,clinical randomized controlled trials (RCTs) of early use of rHu-EPO in premature infants were searched on English databases (PubMed,Embase,Cochrane Collaboration) and Chinese databases (CNKI,SinoMed,Wanfang,and VIP Database).Patients receiving conventional therapy were assigned into control group,and patients receiving both conventional therapy and rHu-EPO were assigned into rHu-EPO group.rHu-EPO group was subdivided into high-dose continuous group and low-dose intermittent group.We retrieved the related literatures,evaluated the quality,and then performed Meta-analysis using RevMan 5.3 software.Result A total of 2 588 patients in 17 studies were included and analyzed.Compared with the control group,Meta-analysis showed that early use of rHu-EPO was more effective in improving NBNA scores at 40 weeks of corrected gestational age (cGA) (MD=2.46,95％CI 1.58～3.33,P＜0.001),and high-dose continuous group was better than low-dose intermittent group (MD=3.19,95％CI 0.45～5.93,P=0.002;MD=2.22,95％CI 1.29～3.16,P＜0.001).Low-dose intermittent use of rHu-EPO significantly increased mental development index (MDI) (MD=6.66,95％CI 0.80～12.51,P=0.010) and psychomotor development index (PDI) (MD=5.77,95％CI 4.00～7.55,P＜0.001),and reduced the incidence of MDI＜70 at cGA 18～22 months (OR=0.42,95％CI 0.27～0.67,P＜0.001).As to the safety and side effects,treatment with rHu-EPO reduced the risk of necrotizing enterocolitis (OR=0.48,95％CI 0.31 ～0.72,P＜0.001) and periventricular leukomalacia (OR=0.52,95％CI 0.35～0.75,P＜0.001)without increasing the risk of bronchial dysplasia,patent ductus arteriosus,retinopathy in prematurity and intraventricular hemorrhage.Conclusion Current evidence shows that the early use of rHu-EPO in premature infants is relatively effective and safe,but multi-center RCTs are still needed.

Objective To investigate the early clinical efficacy of Masquelet membrane induction technique in the treatment of traumatic long bone defects.Methods A retrospective case series study was conducted to analyze the clinical data of 41 patients with traumatic long bone defects admitted to the General Hospital of the Northern Theater Command from January 2012 to April 2017.There were 36 males and five females,aged 15-70 years,with an average of 38.2 years.There were 20 patients with bone defect at the femur,19 at the tibia,one at the fibula,and one at the ulna.All patients received staged treatment using the Masquelet membrane induction technique.In stage Ⅰ surgery,thorough debridement was first performed,and the secretions were taken for bacterial culture.The average bone defect length after debridement was 6.9 cm (2.0-18.5 cm).The bone defect was filled with antibiotic bone cement to induce the biofilm formation.If the postoperative bacterial culture showed positive results,debridement surgery was performed again.Stage Ⅱ surgery was performed after 6-12 weeks.The white blood cell count,C-reactive protein (CRP),procalcitonin (PCT),erythrocyte sedimentation rate (ESR) were measured before the operation.During the operation,bone biopsy was performed,and the bone cement placeholder was completely removed.The autologous cancellous bone and artificial bone were implanted in the bone defect areas,and the induced membrane was sutured.The healing time of bone defects was recorded,and the Paley fracture healing scoring criteria were used to evaluate the limb function.The complications were observed.The inflammatory markers were reviewed at the last follow-up.Results All patients were followed up for 7-36 months with an average of 13.6 months.A total of 37 patients obtained bone healing.The fracture healing rate of stage Ⅰ was 90％,and the healing time was 6-13 months,with an average of 9 months.According to the Paley fracture healing scoring criteria,the results were excellent in 25 patients,good in 10,and fair in two patients,with the excellent and good rate of 85％.In terms of complications,one patient with superficial infection recovered after dressing change,three patients had deep infection,of which one patient was treated with amputation and two received other treatments,and three patients were treated with membrane induction again because of bone resorption.At the last follow-up,there were significant differences between preoperative and postoperative White blood cell count,CRP,PCT and ESR(P ＜ 0.05).Conclusion For traumatic long bone defects,Masquelet membrane induction technique can promote fracture healing,restore limb function and reduce complications.

OBJECTIVE： To study the effects of Aconitum carmichaelii water extract on the expression of 3 kinds of efflux transporters and 3 kinds of tight junction proteins as well as their genes in duodenum tissues of rats. METHODS： Thirty-two SD male rats were randomly divided into normal group， A. carmichaelii water extract low-dose， medium-dose and high-dose groups [0.45， 0.9， 1.8 g/（kg·d）， by crude drug]， with 8 rats in each group. They were given water and relevant liquid 0.1 mL/kg intragastrically for consecutive 7 d. After last administration， the duodenal segments of rats were collected. Western blotting assay was used to detect the expression of efflux transporters as P-glycoprotein （P-gp）， breast cancer resistance protein （Bcrp）， multidrug resistance protein 2 （Mrp2） as well as tight junction proteins as Claudin-1， Occludin and ZO-1. mRNA expression of 6 kinds of proteins relevant gene were determined by qRT-PCR respectively. RESULTS： Compared with normal group， the protein expression of P-gp， Mrp2， Bcrp， Claudin-1， Occludin and ZO-1 in duodenum of rats were increased significantly in A. carmichaelii water extract groups （P＜0.01）. mRNA expression of P-gp in A. carmichaelii water extract groups， mRNA expression of Bcrp in A. carmichaelii water extract low-dose and high-dose groups as well as mRNA expression of Claudin-1 in A. carmichaelli water extract medium-dose and high-dose groups were increased significantly， with statistical significance （P＜0.05 or P＜0.01）. There was no statistical significance in mRNA expression of other genes （P＞0.05）. CONCLUSIONS： A. carmichaelii water extract can up-regulate the expression of 3 kinds of efflux transporters and 3 kinds of tight junction proteins at the level of mRNA and/or protein， thus may interact with other substrates of the aforementioned efflux transporters and drugs with cell bypass pathway as the main transport pathway. In clinical practice， adjustment of dosage may be considered in drug combination.

Objective To evaluate the efficacy and safety of early use of high-dose rHu-EPO in the prevention of anemia in premature infants .Method Available data from clinical randomized controlled trials of early application of high-dose rHu-EPO to prevent premature infant anemia since database building till February 2017 were retrieved, which includes Pubmed, Embase, Cochrane Collaboration and CNKI , SinoMed, Wanfang, VIP.Premature infants less than gestational age of 37 weeks, with routine prevention of anemia as the control group and early combined use of high-dose rHu-EPO≥750 IU/( kg· w) as the treatment group.The relevant data from the literatures obtained were then screened and analyzed by the RevMan 5.3 software.Result A total of 1779 preterm infants from 21 articles including 898 cases of rHu-EPO treatment group and 881 cases in control group.Meta-analysis showed that rHu-EPO treatment group apparently increased the hemoglobin ( MD=27.42, 95%CI 20.01~34.84, P<0.001), hematocrit and reticulocyte count , reduced the blood transfusion. As to the risk of retinopathy of prematurity in rHu-EPO treatment group was significantly higher than that in the control group (RR=1.21, 95%CI 1.01~1.46, P=0.04), and reduced risks of bronchopulmonary dysplasia (RR=0.79, 95%CI 0.46 ~1.35, P =0.39) and patent ductus arteriosus ( RR =0.83, 95% CI 0.68 ~1.01, P=0.07) the difference, however, was not statistically significant, and there was no increasing risks of necrotizing enterocolitis , periventricular leukomalacia , intraventricular hemorrhage .Conclusion Early administration of high-dose of rHu-EPO can effectively prevent anemia in premature infants , but there is an increased risk of retinopathy of prematurity .

Objective To investigate the difference of amplitude of low-frequency fluctuation (ALFF) and fraction of amplitude of low-frequency fluctuation(fALFF) between Alzheimer's disease (AD)patients and normal aging (NA) controls by voxel-based analysis.Methods Thirty-one AD patients and 44 NA controls were enrolled in the study.Blood oxygen level dependent functional (BOLD) EPI data were obtained during resting-state by using 32-channel head coil.Data were realigned,normalized and then smoothed with 8 mm FWHM kernel.Resting-state fMRI toolkit(version 1.6) was used to generate ALFF and fALFF images.Independent two sample t-test was performed with SPM5 to compare ALFF and fALFF of AD and NA controls.Pearson correlation analysis was performed to examine the relationship between MMSE score and ALFF,fALFF parameters.The significance level was set to be uncorrected O.001 on the voxel level and 0.05 on the cluster level.Results AD patients showed increased ALFF in left temporal lobe (0.492 ±0.119) and right cingulated cortex (0.434 ± 0.093) of AD patients,which were 0.443 ± 0.068 and 0.380 ±0.081 in NA controls (t =2.658,2.227,P ＜ 0.05).Decreased fALFF was found in bilateral posterior cingulate cortices (1.167 ± 0.203) and increased fALFF was found in bilateral temporal lobes (left 1.226 ±0.127,right 1.146 ±0.214) with left side dominance,which were 1.453 ±0.269,1.134 ±0.088,1.014 ± O.132 in NA controls (t =5.001,3.695,3.285,P ＜ 0.05).Bilateral temporal ALFF and fALFF correlated with MMSE positively (r =0.768—0.909,P ＜ 0.05) with left dominance.Conclusion AD patients showed increased resting-state functional MRI changes correlated with MMSE score in the temporal lobes with left dominance,which indicated left temporal lobe may be the best location for the observation of disease progression in AD patients.