The peptidyl-prolyl imide bond cis/trans isomerization of Xaa-Pro motif in the peptide and protein plays an important role to influence their conformation and function. Here, a series of model peptides including phosphorylated and its unphosphorylated counterparts were designed and synthesized. Preliminary 1H NMR experiments and molecular dynamics (MD) simulation were used to analyze the peptidyl-prolyl cis/trans imide bond isomerization. The data indicated that the side-chain O-phosphorylation of the Xaa residues preceding proline affected evidently the isomerization and thereby regulated the peptides conformations. The charges of the phosphate moiety as well as their steric effects might be the driving force for the conformational changes of these phosphopeptides.Moreover, the obtained most stable multiple configurations and their statistic cis/trans concentration distribution in MD simulation were basically consistent with the NMR experiments, which demonstrated that phosphorylation increased the cis conformation of the peptide and the maximum cis ratio is given while the phosphate group has no negative charge.

Objective To investigate the effects of Trim34α on the activation of luciferase reporter gene containing NF-κB promoter induced by adaptor proteins TAB2. Methods The total RNA was isolated from HeLa cells. After amplification with RT-PCR, the target sequences were cloned into 5'-Flag-pcDNA3.1 (+) vector. The recombinant vector was confirmed by restriction enzyme digestion, colony PCR and sequencing. It was transfected into HEK293T cells to detected Trim34α expression by Western blot. Simultaneously, the effects of Trim34α on the NF-κB activation induced by TAB2 were determined by dual-luciferase reporter assay. Results Restriction enzyme digestion, colony PCR and sequencing confirmed the vector was constructed successfully, furthermore it expressed Trim34α protein in HEK293T cells. Moreover, trim34α could form high-molecular-weight oligomeric protein, and here we called it trimsome. Interestingly, dual-luciferase assay showed that Trim34α could effectively block TAB2-induced NF-κB activation. Conclusion Trim34α was involved in negative regulation of TAB2-induced NF-κB activation and could form high-molecular-weight oligomer.

Objective To evaluate the method, clinical efficacy and safety of one phase treat-ment of renal calculi associated with pyonephrosis by percutaneous nephrolithotripsy(PCNL) by pneu-matic combined with ultrasonic lithotriptor. Methods Sixty-six cases of renal calculi accompanied with pyonephrosis were treated with PCNL. The renal calyx was punctured under ultrasound gui-dance, then the tract was dilated from F8 to F16 by peel-away vascular access sheathes. After the in-sertion of the flexible sheath, metallic dilator was inserted and the flexible sheath was pulled out. The tract was dilated by metallic sheath to F21 and the operation sheath and nephroseope were placed into working tract. EMS III LithoClast Master was used. Ultrasonic powered lithotriptor probe with suc-tion was used to clear the liquor puris and calculus fragments with low-pressure or no-pressure. The combined pneumatic and ultrasonic powered lithotriptor was used to break and clear the calculi. Re-salts Of the 66 cases, there was no bacteremia or pyaemia intraoperatively and postoperatively. And there was no other severe complication occurred intraoperatively. One phase PCNL was successfully completed in 60 cases. Other 4 cases had residual calculi less than 1.5 em in diameter and received ESWL to break the calculi, 2 cases had bigger residual calculi and accepted second PCNL 1 week after the first intervention. In the follow-up period, the 3 month post-operative serum Cr was 56-203 μmol/L with an average decrease of 40 μmol/L, GFR was 5.0-56.2 ml/min with an average increase of 23.6 ml/min compared with the pre-operative data. At 6 months postoperative serum Cr was 56-158 μmol/L with average decrease of 31 μmol/L, GFR was 5.0-79.2 ml/min with an average in-crease of 30.2 ml/min. Conclusion Application of PCNL in the treatment of patients with renal cal-culi accompanied with pyonephrosis is safe, cost-effective and clinically efficient by pneumatic com-bined with ultrasonic lithotriptor.

OBJECTIVETo explore the relationship between the needling sensation of catgut embedding therapy and the depth of embedded catgut so as to improve the safety of the needle insertion and catgut implantation of the therapy.

METHODSTwenty healthy adults were selected. Under the ultrasound, the structure of the cervical Jiaji (EX-B 2) was observed. In the ultrasound guidance, the catgut was embedded. The two-dimensional imaging method was adopted to observe the anatomic structure and the procedure of needle insertion at the cervical Jiaji (EX-B 2). The high-frequency ultrasound was used to collect the images at Jiaji (EX-B 2) of C5 and determine the depths from the skin surface to the different layers of the point. Additionally, the visual analogue scale (VAS) was adopted to score the needling sensations when the needle inserted at different layers. The persistent sensation duration in the local area was followed continuously.

RESULTSUnder the ultrasound, the anatomic structure and tissue layers of cervical Jiaji (EX-B 2) were displayed clearly. The difference was significant in the average depth from the skin surface to the subcutaneous tissue, trapezius, splenius capitis, semispinalis capitis, semipinalis cervicis, multifidus and vertebral arch between the males and females (all P<0. 01). During the needle insertion, the sensations were apparently different when the implantation went to different layers. The qi arrival presented when the catgut was embedded to the trapezius, splenius capitis, semispinalis capitis, semipinalis cervicis and multifidus. But the distending pain was the most significant when in the myofascial. Commonly, the embedded catgut 2. 5 cm in length may be implanted deeply to the multifidus and the local needling sensation lasted averagely for (72. 0 ± 10. 2) h. Conclusion Under the ultrasound guidance, the depth of embedded catgut is clearly displayed at cervical Jiaji (EX-B 2). The needle insertion and the implanted material are visible, and the relationship between qi arrival and the layer of needle insertion is determined. The accuracy and safety of minimally invasive catgut embedding therapy is improved in the treatment of cervical spondylosis.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Catgut ; utilization ; Female ; Humans ; Male ; Middle Aged ; Neck Muscles ; anatomy & histology ; diagnostic imaging ; Qi ; Sensation ; Spondylosis ; diagnostic imaging ; therapy ; Ultrasonography

Objective To study the effect of TLR4 activation with LPS on BMP9-induced osteogenic differentiation of immortalized mouse embryonic fibroblasts ( iMEFs).Methods The activation of TLR4/NF-κB signaling path-way was detected by ICC.iMEFs were treated with LPS,BAY11-7082,Adnovirus GFP and BMP9.The early osteo-genic differentiation capability of iMEFs was detected by ALP staining and quantitative assay .The later osteogenic differentiation capability was detected by alizarin red S staining .The expression of later osteogenic differentiation marker gene OCN and OPN were detected by PCR and Western blot .The change of p-Smad1/5/8 was detected by Western blot.The expression of Runx2 and Dlx5 were detected by PCR and Western blot .Results LPS can effec-tively stimulate TLR4/NF-κB signaling pathway .TLR4 activation inhibited BMP 9-induced osteogenic differentiation . BMP9-induced osteogenic differentiation related gene and Smad 1/5/8 signaling activation were inhibited by TLR4 activation .The inhibition effect was partly reversed by BAY 11-7082 ( P<0.05 ) .Conclusions TLR4 activation with LPS can inhibit BMP9-induced osteogenic differentiation of iMEFs cells via NF-κB signaling pathway .

Objective:To evaluate the clinical feasibility and image quality of three-dimensional breath-hold gradient and spin-echo (3D BH-GRASE) sequence in magnetic resonance cholangiopancreatography (MRCP).Methods:In this prospective study, 59 patients with clinically suspected pancreaticobiliary duct disease performed MRCP with both 3D BH-GRASE and 3D respiration-triggered turbo spin-echo (3D RT-TSE) sequences on 3.0 T scanner in the Affiliated Zhangjiagang Hospital of Soochow University from November 2017 to December 2018. The overall image quality was scored independently by 3 experienced radiologists based on the visibility of different anatomical features of the pancreaticobiliary duct. For comparing the 2 sequences, the statistical difference in scan time was assessed with a paired t test; while subjective scores, signal-to-noise ratios (SNR), contrast ratios (CR) and contrast noise ratios (CNR) were compared with Wilcoxon signed rank test. Results:The scan time of 3D BH-GRASE sequence was 16.4 s while that of 3D RT-TSE was (258.6±42.2) s. Their difference was statistically significant ( t=44.073, P<0.001), with the scan time for 3D BH-GRASE shortened by 94%. The overall quality scores of 3D BH-GRASE images were better than those of 3D RT-TSE ( Z=-6.595, P<0.001). There was no statistical difference ( P>0.05) in the scores regarding the visibility of the upper, middle and lower parts of common bile duct and the first and second branches of left and right hepatic ducts. For visualizing the bottom, body, neck and duct of gallbladder, the 3D BH-GRASE sequence received a higher score than the 3D RT-TSE sequence ( P<0.001). For displaying the proximal, middle and distal segments of main pancreatic duct, the 3D RT-TSE sequence was scored higher than the 3D BH-GRASE sequence ( P<0.05). There was no significant difference of SNR between the two sequences ( Z=0.403, P=0.687), whereas CR and CNR of 3D RT-TSE MRCP were better than those of 3D BH-GRASE MRCP ( Z=6.215, P<0.001 and Z=3.046, P=0.002, respectively). Conclusion:Under the prerequisite of ensuring image quality, a proper use of 3D BH-GRASE sequence can significantly shorten the scan time and thus greatly improve the working efficiency of MRCP examination.

Objective To evaluate the effect of age factor on the analgesic efficacy of morphine during recovery from remifentanil-based anesthesia in pediatric patients undergoing minor surgery.Methods Fifty pediatric patients of both sexes,aged 3-10 yr,with body mass index ≤ 30 kg/m2,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective tonsillectomy,were divided into preschool group (3-5 yr,n=30) and school-age group (6-10 yr,n =20) according to age.Anesthesia was induced by inhaling 8％ sevoflurane and Ⅳ morphine 0.1 mg/kg.Pediatric patients were mechanically ventilated after tracheal intubation,and end-tidal pressure of carbon dioxide was maintained between 35-45 mmHg.Anesthesia was maintained by inhalation of 2％-3％ sevoflurane and Ⅳ infusion of remifentanil 0.2 μg · kg-1 · man-1.Pain was evaluated using Faces Pain Scale (FPS) and Face Legs Activity Cry Consolability (FLACC) scale during the recovery period in the postanesthesia care unit.When FPS or FLACC scores ≥ 4.morphine 0.05 mg/kg was intravenously injected.When pain was still unrelieved after morphine was given for 2 times (time interval 5 min),fentanyl 1 μg/kg was intravenously injected until FPS and FLACC scores ＜4.The requirement for analgesics and consumption of analgesics (fentanyl consunption was converted into morphine consumption) were recorded.The development of nausea and vomiting,pruritus and respiratory depression was also recorded.Results Compared with preschool group,the requirement for analgesics and consunption of analgesics were significantly decreased during recovery from anesthesia (P＜0.05),and no significant change was found in the incidence of nausea and vomiting in school-age group (P＞0.05).No pruritus or respiratory depression was found in two groups.Conclusion The analgesic efficacy of morphine is affected by age factors during recovery from remifentanil-based anesthesia in pediatric patients undergoing minor surgery,and morphine produces better analgesic efficacy in school-age pediatric patients than in preschool pediatric patients.

Based on the experience of Public-Private Partnership (PPPs) model in domestic and foreign medical field, this study innovatively explored and designed the Build-Transfer-Operate-Transfer (BTOT) operational structure taking the second phase project of ZP Hospital in Pudong New District as a case study.The exploration results showed that the model established the social capital of private non-profit medical institutions by setting up the project company, and used the asset rental and operating management fees in the form of return path.In view of this, the study put forward some relevant countermeasures and suggestions from the aspects of the institution nature and national treatment, property right ownership and risk mitigation, balancing of interests and the cooperation period and solving the core staff etc.All of these measures were taken in order to explore the replicable PPP model in the field of public hospital reform and development.

Objective To observe the expression level of macrophage stimulating protein (MSP) in acute on-chronic liver.failure (ACLF) patients,and to explore the clinical significance and correlation with different immune regulatory factors.Methods The double antigen sandwich enzyme-linked immunosorbent assay method was used to detect MSP in the peripheral blood of 45 patients who were diagnosed with ACLF and 32 cases of chronic hepatitis B (CHB).The MSP levels were compared among ACLF patients with different outcomes,and the MSP level of healthy person was used as control.Meanwhile,liver function and hepatitis B virus (HBV) load were detected,and the expressions of peripheral blood CD4+ interferon (IFN)γ+ (helper T cell 1 Th1),CD4+ interleukin (IL)-4+ (helper T cell 2,Th2),CD4+IL-17+ (helper T cell 17,Th17) and CD4+ CD25+ Foxp3+ (regular T cell,Treg) were measured by flow cytometry.The comparison of means between two samples was done by t test,and oneway ANOVA and linear correlation analysis were also used.Results The serum MSP levels in ACLF patients,CHB group and healthy control were (1.65±0.46) ng/mL,(1.43±0.32) ng/mL and (1.23±0.21) ng/mL,respectively.The serum MSP level in ACLF patients was significantly higher than both CHB patients (t=2.163,P=0.035) and healthy control (t=4.032,P=0.01).The MSP level in ACLF survival group was statistically higher compared with ACLF death group ([2.29 ± 0.42] ng/mL vs [1.42±0.17] ng/mL,t=1.973,P=0.042).Th2,Th17 cells in ACLF group,CHB group and healthy control group were (1.51±0.27) ％ and (1.94±1.02)％,(0.42±0.08)％ and (0.55±0.36)％,(0.23±0.19) ％ and (0.26±0.19) ％,respectively,which were all significantly different (F=7.759 and 37.229,respectively;both P＜0.01).The MSP level was positively associated with the number of Th2 (r=0.386,P=0.032) and Th17 (r=0.644,P=0.000),and the ratio of Th17/Treg (r =0.605,P=0.000);while it was negatively associated with the number of Th1 (r=-0.212),Treg (r=-0.262) and the ratio of Th1/Th2 (r=-0.394) (all P＞0.05).Conclusion MSP is involved in the progress of ACLF,and it may be associated with clinical outcomes and cellular immune imbalance of ACLF patients.

Objective To investigate the effects of dihydropteridine reductase (QDPR) on regulating apoptosis induced by plamitic acid(PA). Methods The transfection of HEK293T cells experiment was divided into 3 groups. A group was the control vector group. B group was the control vector group induced by PA. C group was the recombinant plasmid QDPR group induced by PA. First, control vector and recombinant plasmid QDPR was respectively transfected into HEK293T cells. After 24 h, PA with concentration of 0. 5 mmol/L was added into the medium of above cells. The cells of control vector group, the cells of control vector group induced by PA and the cells of recombinant plasmid QDPR group induced by PA were cultured for another 24 hours. At last, cells were harvested to detect tetrahydrobiopterin (BH4) and reactive oxygen species(ROS) generation, Beclin 1, Caspase 3 and Beclin 2 expression. Results ① After transfection, the recombinant plasmid QDPR was successfully constructed and expressed in cells.② There was no significant difference between A group and B group in BH4 generation. Compared with B group, BH4 generation increased in C group (P < 0. 05).③ ROS generation was increased in B group compared with A group, and decreased ROS generation in C group compared with B group (P < 0. 05).④ Western blot analysis revealed that Beclin 1 and Caspase 3 increased (P < 0. 05 ) while Beclin 2 decreased in B group compared with A group (P < 0. 05). Compared to B group, Beclin 1 and Caspase 3 decreased while Beclin 2 increased in C group (P < 0. 05 ). Conclusion QDPR may regulate apoptosis induced by fatty acids by decreasing the generation of ROS and increasing the level of BH4 and the expression of Beclin 2 associated with anti-apoptosis.