Vortex-assisted dispersive liquid-liquid microextraction based on the solidification of sedimentary deep eutectic solvents for the determination of triazine and phenylurea herbicides in milk samples.

Vortex-assisted dispersive liquid-liquid microextraction based on the solidification of sedimentary deep eutectic solvents was developed and applied to the extraction of triazine and phenylurea herbicides in milk samples. In this study, a series of novel hydrophobic deep eutectic solvents were prepared using tetrabutylammonium chloride as the hydrogen bond acceptor and perfluorooctanol as the hydrogen bond donor, and their structures, viscosities, densities and melting points were determined. The deep eutectic solvent was used as the extraction solvent and dispersed in the sample solution with the assistance of vortex. After extraction, through centrifugation and subsequent cooling in an ice bath, the deep eutectic solvent was solidified and deposited on the bottom of the centrifuge tube. Subsequently, the deep eutectic solvent combined with the target analytes was diluted and used for chromatographic analysis. Some parameters, including the extraction temperature, type and volume of the deep eutectic solvent, amount of NaCl, vortex time and pH of the sample solution, were optimized by the single-factor experiment, Plackett-Burman design and Box-Behnken design. The limits of detection and quantification were in the range of 0.41-0.59 µg L-1 and 1.37-1.95 µg L-1, respectively. The intra-day precision and inter-day precision were in the range of 0.28-2.14% and 2.02-7.99%, respectively. The present method was successfully applied to the determination of triazine and phenylurea herbicides in milk samples.

The mechanism of miR-363-3p/DUSP10 signaling pathway involved in the gastric mucosal injury induced by clopidogrel.

Clopidogrel-induced gastric injury is an important clinical problem. However, the exact mechanism is still unclarified. Increasing evidence indicates that miRNAs may be involved in the pathogenesis of gastric mucosal injury. In this study, the aim was to investigate the role of miR-363-3p in the gastric mucosal injury caused by clopidogrel. MiRNA microarray analysis was performed using paired gastric mucosal in order to find differential expression of miRNAs. The levels of miR-363-3p were examined in gastric mucosal injury caused by clopidogrel. The GES-1 cells were used as a model system, miR-363-3p mimic/inhibitor was transfected into GES-1 cells, then GES-1 cells were treated with clopidogrel. The levels of miR-363-3p and DUSP10 were examined in GES-1 cells using quantitative real-time PCR (qRT-PCR). CCK-8 assay and flow cytometry analysis were used to detect cell proliferation and apoptosis, respectively. Western blot assay was used to measure the protein levels of DUSP10. The interaction between miR-363-3p and DUSP10 was determined by luciferase reporter assay. MiR-363-3p was selected as a differentially expressed miRNA. The expression of miR-363-3p in gastric mucosal injury caused by clopidogrel was higher than that in normal samples. Also, depletion of miR-363-3p increased the proliferation of GES-1 cells and reduced the apoptosis. Luciferase-reporting assay results confirmed that DUSP10 was one of the target genes of miR-363-3p. DUSP10 inhibited apoptosis in GES-1 cells treated by clopidogrel. Moreover, DUSP10 knockdown abrogated the inhibitory effects on apoptosis in GES-1 cells mediated by miR-363-3p inhibitor. Knockdown of miR-363-3p increased the proliferation and reduced the apoptosis by targeting DUSP10 in GES-1 cells treated by clopidogrel, indicating that miR-363-3p may be a potential therapeutic target for gastric mucosal injury caused by clopidogrel.

Method for the detection of submucosal tumors of the esophagus in submucosal tunnel under the preoperative identification / 中国内镜杂志

Objective To investigate the preoperative identification of esophageal submucosal tumor by endoscope. Methods 40 patients of esophageal submucosal tumors with lesions range from 1.0 to 2.0 cm from March 2012 to August 2016 were randomly divided into A, B groups. Patients in group A underwent submucosal tunneling endoscopic resection (STER) 3.0 cm from the lesions, while patients in group B first underwent submucosal injection of saline to mark the lesions, then perform STER in the same way. Then record the time of checking. Results The average time of group A was (420.0 ± 25.0) s, the average time of B group was (300.0 ± 25.0) s, there was statistical differences between the two groups. Conclusion Preoperative identification of the lesions before STER holds more advantages.

Role of topical tranexamic acid plus cocktail analgesic in reducing blood loss during total knee arthroplasty / 中华创伤杂志

Objective To investigate the effect and safety of topical tranexamic acid (TXA) plus cocktail analgesic for reducing blood loss during total knee arthroplasty (TKA).Methods A prospective case control study was made on 60 patients scheduled to undergo TKA because of knee injuries between August 2015 to June 2016.There were 13 males and 47 females,with the mean age of 65.5 years (range,51-80 years).Traumatic arthritis occurred in 44 patients and degenerative arthritis in 16 patients.The patients were assigned to separate cocktail analgesic group (Group A,n =30) and topical TXA plus cocktail analgesic group(Group B,n =30),according to the random number table.Patients in Group A received multiple-point intra-articular cocktail analgesic injection before implantation of the prosthesis in TKA.While patients in Group B received multiple-point intra-articular TXA plus cocktail analgesic injection before implantation of the prosthesis.Between-group differences were compared with respect to intraoperative blood loss,hemoglobin change (Hb),haematocrit (Hct),postoperative drainage,total blood loss,hidden blood loss,blood transfusion rate,Hospital for Special Surgery (HSS) score,incidence of deep venous thrombosis (DVT) and other complications.Results All patients were followed up for 3 months.Perioperative Hb reduction in Group B was 18.5 (13.0,26.0) g/L,less than 23.0 (21.0,35.5) g/L in Group A (P ＜ 0.05).Hct was reduced by 5.6 (4.1,7.8) ％ in Group B,while 7.2 (6.1,10.7) ％ in Group A (P ＜ 0.05).Postoperative drainage volume,total blood loss and occult blood loss in Group B were 105.0(60.0,223.8) ml,596.0(426.1,795.3) ml,422.3 (228.9,624.0) ml respectively,decreased compared to Group A [162.5 (118.8,245.0) ml,788.3 (583.0,1 082.4) ml,603.2 (435.2,884.7)ml respectively] (P ＜0.05).There were no significant differences in intraoperative blood loss,blood transfusion rate,HSS score and DVT incidence between the two groups (P ＞0.05).Conclusion Topical TXA plus cocktail analgesic can reduce blood loss during perioperative period in TKA,without increasing the risk of DVT.

Estrogen promotes spontaneous repair of osteochondral defect via inhibition of osteoclast activity / 中华创伤杂志

Objective To investigate the role of estrogen in spontaneous repair of osteochondral defect,so as to provide guidance for the application of estrogen in tissue engineering.Methods Seventytwo healthy adult female SD rats (4-month-old) were assigned to negative control group (group A,n =24),ovariectomized group (group B,n =24) and ovariectomized + estrogen treatment group (group C,n =24) according to the random number table.Rats in group C were intraperitoneally injected 0.1 mg/kg estradiol benzoate once per week after operation.An osteochondral defect model (diameter:2 mm,height:1.5 mm) was established in femoral trochlea of rats 4 weeks after operation.At 2,10 and 20 weeks after the modeling,8 rats from each group were sacrificed.Femoral condyles were collected to measure bone volume fraction (BVF),number of bone trabeculae (Tb.N),trabecular thickness (Tb.Th) and trabecular spacing (Tb.Sp) in the osteochondral defect area by micro-CT scanning.Morphological changes were observed by HE staining,histomorphological changes and cartilage regeneration by safranine O-fast green staining,osteoclast count by tartrate-resistant acid phosphatase (TRAP) staining,distribution of matrix metalloproteinase-9 (MMP-9) by immunohistochemistry and expressions of osteoprotegerin (OPG),receptor activator of nuclear factor-κB ligand (RANKL) and MMP-9 by real-time-PCR.Results Compared to group B,increased BVF and Tb.N and decreased Tb.Sp were observed in groups A and C at each time point.More bone matrix formations were observed in groups A and C than in group B at 2 weeks,cysts (also known as cystic cavities) in regenerated subchondral bone and cracks in subchondral bone plate were observed in group B at 10 and 20 weeks,and proliferation of osteoclasts was active within lesions in group B at 10 and 20 weeks.Compared to group B,lowered levels of RANKL and MMP-9 (P ＜ 0.05) and significantly increased level of OPG (P ＜ 0.01) were observed in groups A and C at each time point,and osteoclast count in groups A and C was lowered at 10 and 20 weeks (P＜0.05).No regeneration of cartilage layer was observed in all groups.Conclusions Excessive proliferation of osteoclasts results in formation of cystic cavities and cracks.Estrogen improves the speed and quality of bone repair by regulating activities of osteoblasts and osteoclasts,so it can be used to treat osteochondral defect in tissue engineering.

The construction of a tissue-engineered tendon mimicking the transitional architecture at the ligament-bone interface in rabbit / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate a method that constructing a tissue-engineered tendon with a continuous and heterogeneous transition region.</p><p><b>METHODS</b>Fibroblasts derived from rabbit epithelial tissue were cultured in vitro and collagen gel was prepared. The experimental groups were scaffold only group, fibroblasts+ chondrocytes group (Fb+ CC group), fibroblasts+ osteoblasts group (Fb+ OB group), fibroblasts+ chondrocytes+ osteoblasts group (Fb+ CC+ OB group). Heterogeneous cell populations(fibroblasts, chondrocytes and osteoblasts) with collagen gel were seeded within three predesigned specific regions (fibrogenesis, chondrogenesis, and osteogenesis) of decellularized rabbit achilles tendons to fabricate a stratified scaffold containing three biofunctional regions supporting fibrogenesis, chondrogenesis, and osteogenesis. The tests of morphology, architecture and cytocompatibility of the scaffolds were performed. Gradient tissue-specific matrix formation was analysed within the predesignated regions via histological staining and immunofluorescence assays.</p><p><b>RESULTS</b>The HE staining and scanning electron microscopy analysis demonstrated that no major cell fragments or nuclear material was evident, and increased intra-fascicular and inter-fascicular spaces were found, the cytocompatibility of the scaffolds showed that the numbers of viable cells on the scaffold surfaces increase steadily, no significant differences were found between the scaffold only containing ordinary culture medium and scaffold containing gel groups. Histological staining and immunofluorescence assays demonstrated that the cartilage-related markers (GAG, COL2A1) were found only in the chondrogenesis region, but bone-related proteins only in the osteogenesis region of bone tunnel, and fibrosis was remarkable for the fibrogenesis region in the joint cavity. The transitional architecture with ligament-fibrocartilage-bone was constructed in the ligament-bone tunnel interface.</p><p><b>CONCLUSIONS</b>A transitional interface (fiber-fiberocartilage-bone) could be replicated in a decellularized tendon through stratified tissue integration in vitro. The cell-tendon complex offers the advantages of a multi-tissue transition involving controlled cellular interactions and matrix heterogeneity.</p>

Early complications of cervical spine surgery in 96 patients / 中国基层医药

Objective To evaluate the early complications of cervical spine surgery .Methods We retro-spectively analyzed 96 cervical spine surgery patients in our department ,including 56 cervical spondylotic myelopathy , 21 cervical fracture and/or dislocation ,11 cervical spine tumor ,5 atlantoaxial dislocation ,3 Chiari malformation .By analyzing causes of complications ,the countermeasures were developed .Results 27 patients had complications .The major complications were:death in 1 case,incision hematoma in 2 cases,incision infection in 4 cases,spine cord inju-ry or nerve root injury in 3 cases,cerebrospinal fluid leakage in 3 cases,superior laryngeal nerve and recurrent laryn-geal nerve injury in 4 cases,pulmonary infection in 5 cases,urinary tract infection in 4 cases.There were no esophage-al fistula and vertebral artery injury in these patients .The incidence rate in anterior ,posterior,anterior combined with posterior surgery was 24.6%(14/57),36.8%(7/19),40.0%(6/15) respectively.Conclusion Cervical spine surgery is likely to get early complications .Adequate preoperative preparation and improving operative techniques , timely and correctly handle the complications could reduce complications and improve cure rate .

The study on the mechanim of clopidogrel in human gastric epithelial GES-1 cell line injury / 中华消化杂志

ObjectiveTo explore the mechanism of clopidogrel in human gastric epithelial cell line (GES-1) injury.MethodsSet up GES-1 cells monolayer culture model.Then the GES-1 cells were divided into negative control group,U0126 intervented group,clopidogrel intervented group and combined intervented group (U0t26 treated firstly then clopidogrel intervented).The cell proliferation and apoptosis in each group was examined by methyl thiazolyl tetrazolium (MTT) assay and Flow cytometry.TheexpressionofphosphorylatedERK1/2ineachgroupwasdetectedby immunocytochemistry method,and the expression quantity of phosphorylated ERK1/2 in each group was measured by western blot.ResultsThe result of MTT assay showed that compared with negative control group,the proliferation of GES-1 cells was inhibited in U0126 group,clopidogrel group and combined intervented group,and the inhibition percentage was 21.8％ ±2.7％,46.3％ ± 3.4％ and 82.9 ％ ± 0.8 ％ respectively ( F=615.556,P =0.000 ).The result of immunocytochemistry indicated that the expression of p-ERK in U0126 group,Clopidogrel group and combined intervented group decreased compared with negative control group,which was 10.80±1.64,7.20± 1.64,4.40±0.89and 1.40±0.55 respecitively (F=49.426,P=0.000).The result of western blot and immunocytochemistry was of the same trend.Conclusion In GES-1 cell model,clopidogrel may injureGES-1 cells through MAPK/EPK signal transduction pathway.

Injured bile duct repair and liver disease / 中华肝胆外科杂志

The wound healing research in the biliary tree lag significantly behind similar studies on the skin and gastrointestinal tract. This is at least partly attributable to the difficulty to study on the biliary tract. But clinical relevance, more interest in biliary epithelial cell (BEC) pathophysiology, and widespread availability of BEC cultures are factors reversing this trend. In the extra-hepatic biliary tree,ineffectual wound healing, scarring and stricture development are pressing issues. In the smallest intra-hepatic bile ducts either impaired BEC proliferation or an exuberant response can contribute to liver disease. Chronic inflammation and persistent wound healing reactions in large and small bile ducts often lead to hepatic cirrhosis and liver cancer.

Methodology of transfecting gene into liver graft mediated by adeno-associated virus vector / 中华消化外科杂志

Objective To investigate the effective route and proper method in transfecting gene into liver graft mediated by adeno-associated virus vector.Methods Three routes including hepatic artery,portal vein and hepatic artery+portal vein,and 3 methods,i.e.routine,circulation and clamping were employed for infusion.The best infusion route and method of gene transfection into liver graft were determined by observing the color change of liver and detecting liver function and transfoetion rate of liver cells.The safety of these methods was evaluated.Results In all the infusion procedures,the color of the liver grafts turned from red to white,no apparent color differenee of the livers and no enlargement nor mottling were observed under surgical microscope.The liver color was back to normal immediately after blood flow was restored.No significantly statistical differences of the ALT values were observed among all the groups(F=0.343,1.265,0.055,P＞0.05).Adeno-associated virus vectors coding for the enhanced green fluorescence protein(AAV2-EGFP)were successfully transfected into liver cells by the 3 infusion routes 1 week later,and the difierences of transfection rates via the 3 routes had no statistical significance(F=0.080,0.091,0.045,P＞0.05).The transfoction rate of AAV2-EGFP was the highest at any time points when using the clamping method,and then followed by circulation method and routine method,with statistical differenee(F=3.880,2.976,5.129,P＜0.05).The transfection rates of AAV2-EGFP were increased progressively and peaked at the 6th week,and then they were decreased gradually.Conclusions Infusion via hepatic artery is the effective route for gene transfection and clamping the vessels can elevate the transfection rate of AAV2-EGFP.All procedures were performed without detectable liver injury.The transfection of gene into liver graft mediated by adeno-associated virus vector is a slow and persistent process.

Genotype identification of Orientia tsutsugamushi isolated from Nan Peng Lie Islands in China / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To identify genotype of eight strains of Orientia tsutsugamushi (O. tsutsugamushi) isolated from Nan Peng Lie Islands in China and establish tsutsugamushi disease nature foci for this region.</p><p><b>METHODS</b>The nested polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) were used. Three primers were selected from the DNA sequence of the gene encoding type-specific 56-kDa protein of the Karp strain. The positive products were digested by Hine II and Pst I, meanwhile profiles specific to each strain were analyzed.</p><p><b>RESULTS</b>Three genotypes of O. tsutsugamushi including Karp, Kato and a new strain existed on Nan Peng Lie Islands.</p><p><b>CONCLUSION</b>Nan Peng Lie Islands is tsutsugamushi disease nature foci.</p>