OBJECTIVE To investigate the effects and mechanism of galangin （GAL） on hepatocyte apoptosis in rats with obstructive jaundice （OJ） based on the Janus kinase 2 （JAK2）/signal transduction and activator of transcription 3 （STAT3） signaling pathway. METHODS Taking male SD rats as the object， the OJ model was established by double ligation of common bile duct， and 48 rats with successful modeling were randomly separated into OJ model group （model group）， low-dose GAL group （GAL-L group）， high-dose GAL group （GAL-H group） and high-dose GAL+JAK2 activator colivelin group （GAL-H+ colivelin group）， with 12 rats in each group； another 12 SD rats with laparotomy/abdominal closure without ligation were selected as sham operation group （sham group）. Each administration group was given relevant medicine intragastrically and/or intraperitoneally， once a day， for 7 consecutive days. After the last medication， the morphology of liver tissue in rats was observed； the serum levels of total bilirubin （TBIL）， direct bilirubin （DBIL）， alanine transaminase （ALT） and γ -glutamyltransferase （GGT）， as well as the levels of superoxide dismutase （SOD） and malondialdehyde （MDA） in mail：guoweishengjcy@126.com liver tissue were detected. The apoptotic rate of liver tissue cells， the expression levels of signaling pathway-related proteins （phosphorylated JAK2， JAK2， phosphorylated STAT3， STAT3） and apoptosis-related proteins [B cell lymphoma 2 （Bcl-2）， Bcl-2 related X protein （Bax）] were determined. RESULTS Compared with sham group， congestion of liver sinusoids， damage to liver lobules， disordered arrangement and swollen morphology of liver cells， the disappearance of nucleoli， and significant infiltration of inflammatory cells and fibrous tissue proliferation were observed in model group； the serum levels of TBIL， DBIL， ALT and GGT， the level of MDA in liver tissue， the apoptosis rate of liver cells， the protein expression of Bax， and the protein phosphorylation levels of JAK2 and STAT3 in liver tissue of model group were increased significantly （P＜0.05）； the level of SOD and the protein expression of Bcl-2 in liver tissue were decreased significantly （P＜0.05）. Compared with the model group， the pathological injuries of liver tissue were relieved in GAL-L group and GAL-H group， all quantitative indicators had significantly improved， and the effect of GAL-H group was more significant （P＜ 0.05）. Colivelin could significantly reverse the improvement effects of GAL on liver injury and related indicators of OJ rats （P＜ 0.05）. CONCLUSIONS GAL may inhibit liver cell apoptosis in OJ rats， improve liver function and alleviate oxidant stress， the mechanism of which may be associated with inhibiting JAK2/STAT3 signaling pathway.

Objective:To study the application of 3D printing technology in multi-center fenestrated/branched endovascular repair (F/B-EVAR) for endovascular repair of abdominal aortic diseases.Methods:From Feb 2018 to Mar 2023, The clinical and followup data of 316 cases of abdominal aortic lesions undergoing repair with F/B-EVAR at 69 medical centers nationwide using 3D printing technology to guide physician-modified stent graft were retrospectively analyzed.Results:The mean follow-up time of the patients was 23 months (2-60 months), and 24 cases were lost to follow up, the follow-up rate was 92.4% (292/316), the mean postoperative hospitalization time was (8.2±4.9) days. A total of 944 main abdominal branch arteries were reconstructed. Intraoperative reconstruction of 11 branches failed, with a success rate of 98.8% (933/944). Within 30 days after surgery, 8 patients died (2.5%), and 6 patients died during follow-up, a total of 14 patients died (4.4%). There were 11 cases (3.5%) of spinal cord ischemia and no patient suffered from permanent paraplegia. There were 19 patients (6.0%) with postoperative renal function injury. Internal leakage was found in 26 patients, and the rate of internal leakage was 8.2%.Conclusion:3D printing technology can accurately locate the location of branch arteries, simplifing the surgical process, shortening the learning curve , and improving clinical efficacy.

Objective:To observe the surgical methods and clinical efficacy on reconstruction of digital flexion function using transfer of modified free gracilis myocutaneous flap.Methods:Between March 2014 and August 2022, 7 male patients, aged between 23 and 38 (average age 28) years old, were treated by reconstruction of forearm flexor function using modified free gracilis myocutaneous flap transfer to overcome the dysfunction in the Department of Hand and Foot Surgery, Zhejiang Rongjun Hospital. After the scar excision, the defects in forearm ranged from 15.0 cm × 4.5 cm to 28.0 cm × 6.5 cm, with flap excision areas of 17.0 cm × 5.5 cm to 30.0 cm × 8.0 cm. The nerve carried by the myocutaneous flap was anastomosed with the musculi branch of the median nerve, and the perforator artery and vein of the gracilis muscle were anastomosed with the brachial artery or the branch of ulnar artery and radial artery. Six patients had the flap donor sites directly closed primarily. A medium-thickness skin graft was taken from the ipsilateral groin area in 1 patient, to cover the remaining wound after the primary closure for wound reduction. Postoperative clinical efficacy was monitored through the follow-ups via visits of outpatient clinic, telephone calls or WeChat interviews.Results:After surgery, 1 patient experienced a vascular compromise of the flap, which was resolved after reanastomosis. One patient had a necrotic in distal flap about 2.0 cm×1.0 cm in size, which healed after dressing change. All other 5 flaps survived successfully. The postoperative follow-up ranged from 12 to 36 months, at 18 months in average. The flaps had good appearance and texture. Protective sensation recovered to varying degrees at 6 months after surgery. At 2 years after surgery, all of 7 patients showed significant improvement in hand and forearm appearance, along with restored finger flexion, grasping and partial hand function. According to the Evaluation Standard of Upper Limb Functional of Hand Surgery of Chinese Medical Association, flexion function recovered to excellent in 1 patient, good in 3 patients and fair in 3 patients. In the donor sites, there were only linear scars or mild pigmentation without significant loss of function.Conclusion:A modified free gracilis myocutaneous flap transfer is used to reconstruct digit flexion function. The procedure is safe and reliable, with a quick and relatively postoperative and satisfactory recovery in forearm flexion function, hence makes it an ideal surgical technique for reconstruction of digit flexion function.

Objective To evaluate the clinical effect of treatment for deep burn wound on finger by proper digital artery flap.Methods From March 2013 to October 2016, 24 patients with deep burn wound on finger were treated by proper digital artery flap.Postoperative observation included wound repair, flap survival, complications and functional recovery of fingers.Results All the 24 flaps survived and no necrosis happened.The marginal abnormal circulation of flap occurred in only 5 cases, which cured by dress changing.All flaps kept well in contour, skin color, temperature and texture.Movement function of donor and recipient fingers was nearly normal.Conclusion Proper digital artery flap avoided the deficiencies distant pedicled flap, so it is a favorite choice for digital soft tissue defect caused by deep burn injury.

OBJECTIVETo explore the immuno-effect of plasmacytoid dendritic cells (pDC) on bacteria infection induced spontaneous remission (SR) of leukemia.

METHODSBoth pDC and myeloid dendritic cells (mDC) were isolated and purified from leukemic patient with SR and healthy donor by combination of immunomagnetic beads and flow cytometry. pDC were cultured in RPMI1640 medium and stimulated with different bacteria. The T cells proliferation was detected by MTT, and cytokine production by ELISA kits.

RESULTSThe human bacterial pathogen Staphylococcus aureus and Pseudomonas aeruginosa stimulation for 48 h resulted in the maturation of pDC with production of high quantity of IFN-α at (15.34 ± 2.91) ng/ml and (10.38 ± 1.41) ng/ml, respectively, comparing with that of negative group at (1.36 ± 0.13) ng/ml (P<0.01). Activated pDC could promote the differentiation of naive CD4⁺ T cells to Th1 cells with secretion of IFN-γ at (2.16 ± 0.37) ng/ml and (2.73 ± 1.11) ng/ml, respectively, comparing with that of positive control at (2.55 ± 0.23) ng/ml (P > 0.05). Activated pDC showed higher T cell stimulatory capacities [proliferation index (PI) was 4.36 and 4.05, respectively] than that of non-activated pDC (PI was 1.23 and 0.13, respectively) (P < 0.01).

CONCLUSIONStaphylococcus aureus and Pseudomonas aeruginosa activated pDC may play a key role in SR of leukemia following severe infections.

CD4-Positive T-Lymphocytes ; Dendritic Cells ; immunology ; Flow Cytometry ; Humans ; Interferon-alpha ; Leukemia ; diagnosis ; immunology ; Lymphocyte Activation ; Pseudomonas aeruginosa ; immunology ; Remission, Spontaneous ; Staphylococcus aureus ; immunology

A Cleanert Alumina-N-SPE column (0.5 g/6 mL) chromatograpy with 5 mL of chloroform-methanol (7: 3) as eluent, instead of aluminum oxide column (100-200 mesh, 5 g, 1 cm) chromatograpy eluted successively with chloroform and the chloroform-methanol (7:3) (20 mL each), was applied to enrich matrine and oxymatrine in Sophora flavescens. Also, the optimization of the HPLC determination conditions with acetonitrile-ethanol absolute-3% phosphoric acid solution (84: 6: 10) as mobile phase, instead of acetonitrile-ethanol absolute -3% Phosphoric acid solution (80: 10: 10) recorded in Chinese Pharmacopoeia 2010 Edition, was more suitable for determination of matrine and oxymatrine in S. flavescens. This method has advantage of reducing sample handling time and solvent volume and increasing the accuracy and feasibility, which can simplify the procedure for determination of matrine and oxymatrine in S. flavescens.
Alkaloids ; analysis ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Quinolizines ; analysis ; isolation & purification ; Solid Phase Extraction ; methods ; Sophora ; chemistry

The newly emerged Middle East respiratory syndrome coronavirus (MERS-CoV) is a highly pathogenic respiratory virus with pathogenic mechanisms that may be driven by innate immune pathways. The goal of this study is to characterize the expression of the structural (S, E, M, N) and accessory (ORF 3, ORF 4a, ORF 4b, ORF 5) proteins of MERS-CoV and to determine whether any of these proteins acts as an interferon antagonist. Individual structural and accessory protein-coding plasmids with an N-terminal HA tag were constructed and transiently transfected into cells, and their native expression and subcellular localization were assessed using Wes tern blotting and indirect immunofluorescence. While ORF 4b demonstrated majorly nuclear localization, all of the other proteins demonstrated cytoplasmic localization. In addition, for the first time, our experiments revealed that the M, ORF 4a, ORF 4b, and ORF 5 proteins are potent interferon antagonists. Further examination revealed that the ORF 4a protein of MERS-CoV has the most potential to counteract the antiviral effects of IFN via the inhibition of both the interferon production (IFN-β promoter activity, IRF-3/7 and NF-κB activation) and ISRE promoter element signaling pathways. Together, our results provide new insights into the function and pathogenic role of the structural and accessory proteins of MERS-CoV.
Cell Line ; Coronavirus ; genetics ; pathogenicity ; Genes, Viral ; Humans ; Interferons ; antagonists & inhibitors ; Open Reading Frames ; Recombinant Proteins ; genetics ; metabolism ; Viral Matrix Proteins ; genetics ; metabolism ; Viral Regulatory and Accessory Proteins ; genetics ; metabolism ; Viral Structural Proteins ; genetics ; metabolism

Objective Using M2-Gi1α fusion protein expressed by baculovirus-Sf9 cell system to find the specific ligand for M2 receptor and detect the interaction of the two parts of the fusion protein.Methods The fused M2-Gi1α cDNAs were generated in a two-step PCR and then expressed in Sf9 cells.[3H]QNB and[35S]GTPγS binding experiments were employed to study the function of M2-Gi1α fusion protein.Results The expression level of M2-Gi1α fusion protein was 8.44±0.39 nmol·g-1 protein.The affinity of GDP to the Gi1α part changed under the affection of different ligands.The IC50 value in the appearance of acetylcholine,oxotremorine,arecoline,atropine,fangchinoline,levitimide were 21.35 μmol·L-1,23.86 μmol·L-1,11.91 μmol·L-1,0.13 μmol·L-1,1.05 μmol·L-1,1.75 μmol·L-1,and 2.5 μmol·L-1 when there was no ligand.Conclusion The M2-Gi1α fusion protein expressed in baculovirus-Sf9 cell system has pharmacological specificity for M2 receptor and the efficient coupling function between the two parts.The M2-Gi1αfusion protein is a helpful tool for detecting the new specific ligands of the M2 receptor.

Aim The m_5AChR-G_ 11? fusion protein was expressed by baculovirus-Sf9 cells system, then using it to identify the specific agonists and antagonists for m_5AChR via detecting the affinity of GDP and m_5AChR-G_ 11?. Methods The m_5AChR-G_ 11? fused cDNAs were generated via a two-step PCR protocol and inserted into pBacPAK9 virus vector. We expressed m_5AChR-G_ 11? fusion protein and m_5AChR protein using baculovirus-Sf9 cell system. [ 3H]QNB and [ 35S]GTP?S binding tests were performed to detect the expressional level of receptor proteins and determine the affinity of GDP and m_5AChR-G_ 11? fusion protein. Results The expression level of m_5AChR-G_ 11? was (47.6?3.2) nmol?g -1 protein. The affinity of GDP to G_ 11? partner changed in the presence of different muscarinic ligands. IC_ 50 values of GDP in the presence of ACh, YM796, Oxotremorine, Methixene, Dextimide and atropine were 128.0, 72.1, 68.5, 16.2, 14.9 and 9.7 ?mol?L -1 respectively, and that in the absence of muscarinic ligand was 20.8 ?mol?L -1. Conclusion The m_5AChR-G_ 11? fusion protein has the pharmacological specificity of M_5 receptor and the efficient coupling interaction of the two partner. Affinity of GDP to ligand bound m_5AChR-G_ 11? fusion protein represents the species of muscarinic ligands. ACh is a full agonist for m_5AChR-G_ 11? fusion protein, YM796 and oxotremorine are partial agonists, while methixene, dextimide and atropine are antagonists.

Aim To generate m3AChR-G11 fusion protein in baculovirus-Sf9 cells and test the couping function,the interation and the influence factors be-tween m3AChR and G11 protein,as well as screen the specific ligands for m3AChR. Methods m3AChR-G11 fused DNA was generated through a two-step PCR and then expressed in Sf9 cells to produce fusion protein. The total concentration for membrane protein was de-tected by BCA method,[3H]QNB and [35 S]GTP?S binding experiment as perfomed to study the function of m3AChR-G11 fusion protein. Results The expression level of m3AChR-G11 was 7. 76 ? 10 -9 mol?g -1. The affinity of GDP to G11 partner changed in the presence of different muscarinic ligands. IC50 values of GDP in the presence of ACh,Pilo,CCh,MCN-A-343,Atro,4-Damp and Dafi were 82. 2,93. 70,12. 10,14. 30, 1. 93,1. 37,0. 72 ? 10 -6 mol ? L -1 respectively,and that in the absence of muscarinic was 1. 99 ? 10 -6 mol ?L-1.Concluslons The m3AChR-G11 fusion protein has the pharmacological specificity of m3 receptor and the efficient coupling interaction of the two partners. Affinity of GDP to ligand-bound fusion protein represents the species of muscarinic ligands. This is helpful in screening and detecting the new specific ligands to muscarinic receptors.