Role of HBV DNA polymerase in mediating the immune escape of tumor cells in HBV-related hepatocellular carcinoma / 临床肝胆病杂志

ObjectiveTo determine whether HBV DNA polymerase is associated with T-cell failure and thus mediates the immune escape of HBV-related hepatocellular carcinoma （HCC） tumor cells， and to investigate the specific molecular mechanisms. MethodsLiver cancer cell lines Huh7 and HepG2 stably transfected with HBV DNA polymerase expression plasmid with Flag （Flag-HBV-P） and intercellular adhesion molecule-1 （ICAM1） were co-cultured with Jurkat cells， and MTT assay， qRT-PCR， and ELISA were used to measure Jurkat cell proliferation， activation （CD69 expression）， and secretion of the cytokine IFN-γ. RNA-seq was used to screen for differentially expressed immune-associated molecules between stably transfected cell lines and control cells， and mRNA half-life and protein half-life assays were used to determine the specific levels of the immune-associated molecules that were affected by HBV DNA polymerase. Related websites were used to predict the transcription factors that may bind to the promoter region of this immune-associated molecule， Western blot was used to verify the effect of transcription factors on the immune-associated molecule， and rescue experiment was used to determine whether HBV DNA polymerase affects the expression level of the immune-associated molecule through this transcription factor. The independent-samples t test was used for comparison between two groups. ResultsThe experimental group had significant reductions in Jurkat cell proliferation， activation， and cytokine secretion compared with the control group （all P<0.01）. Compared with the control group， the experimental group （Huh7 and HepG2 cell lines） had significant reductions in the mRNA and protein expression levels of ICAM1 （all P<0.01）. Website prediction identified the ICAM1 promoter and preliminarily highlighted NFKB1， RELA， and STAT3. Compared with the control group， the experimental group （Huh7 and HepG2 cell lines） had a significant reduction in the protein expression level of p65 （all P<0.01）. After p65 overexpression， there was a significant increase in the protein expression level of ICAM1， and after the expression of p65 was reduced， there was a significant reduction in the protein expression level of ICAM1 （all P<0.01）. In the rescue experiment， there was no significant difference in the protein expression level of ICAM1 between the control group and the experimental group after p65 overexpression （all P>0.05）. After the overexpression of ICAM1， there were no significant differences in the proliferation， activation， and cytokine secretion of Jurkat cells between the control group and the experimental group （Huh7 and HepG2 cell lines） （all P>0.05）. ConclusionHBV DNA polymerase downregulates the level of ICAM1 to mediate HCC immune escape by inhibiting the expression of p65 in NF-κB.

Solution structure of the RNA recognition domain of METTL3-METTL14 N-methyladenosine methyltransferase

N-methyladenosine (mA), a ubiquitous RNA modification, is installed by METTL3-METTL14 complex. The structure of the heterodimeric complex between the methyltransferase domains (MTDs) of METTL3 and METTL14 has been previously determined. However, the MTDs alone possess no enzymatic activity. Here we present the solution structure for the zinc finger domain (ZFD) of METTL3, the inclusion of which fulfills the methyltransferase activity of METTL3-METTL14. We show that the ZFD specifically binds to an RNA containing 5'-GGACU-3' consensus sequence, but does not to one without. The ZFD thus serves as the target recognition domain, a structural feature previously shown for DNA methyltransferases, and cooperates with the MTDs of METTL3-METTL14 for catalysis. However, the interaction between the ZFD and the specific RNA is extremely weak, with the binding affinity at several hundred micromolar under physiological conditions. The ZFD contains two CCCH-type zinc fingers connected by an anti-parallel β-sheet. Mutational analysis and NMR titrations have mapped the functional interface to a contiguous surface. As a division of labor, the RNA-binding interface comprises basic residues from zinc finger 1 and hydrophobic residues from β-sheet and zinc finger 2. Further we show that the linker between the ZFD and MTD of METTL3 is flexible but partially folded, which may permit the cooperation between the two domains during catalysis. Together, the structural characterization of METTL3 ZFD paves the way to elucidate the atomic details of the entire process of RNA mA modification.

Expression of PSAT1 in pancreatic cancer tissues and the mechanism underlying PSAT1-mediated cell proliferation and invasion / 中国肿瘤临床

Objectives: To investigate the expression of phosphoserine aminotransferase 1 (PSAT1) in pancreatic cancer tissues, and its potential role in pancreatic cancer. Methods: The expression of PSAT1 in 98 human pancreatic cancer tissues, which were collected from the People＇s Hospital of Guizhou, between July 2013 to July 2017, and the corresponding adjacent normal tissues was analyzed by immunohistochemical staining. Additionally, the relationship between the expression of PSAT1 and the clinicopathological parame-ters, overall survival (OS), and disease-free survival (DFS) of patients with pancreatic cancer was evaluated. The human pancreatic can-cer cell lines, BxPC-3 and SW1990, were transfected with PSAT1-siRNA, to investigate the effect of PSAT1 knockdown on cell prolifera-tion, migration, and invasion. Additionally, we performed Western blot to assess the expression of PI3K/Akt/mTOR-related proteins in PSAT1-knockdown cells. Results: The percentages of PSAT1-positive cells in pancreatic cancer and adjacent non-tumor tissues were 69.4% (68/98) and 5.0% (5/98), respectively, indicating a significantly higher expression of PSAT1 in pancreatic cancer tissues com-pared to adjacent non-tumor tissues (P<0.05). The increased expression of PSAT1 in pancreatic cancer tissues correlated with lymph node metastasis and TNM stage. Kaplan-Meier analysis suggested that a high expression of PSAT1 correlated with a poor OS and DFS compared to a low expression of PSAT1 (P<0.05). Cox regression analysis showed that the expression of PSAT1 is an independent prog-nostic marker for OS and DFS in pancreatic cancer patients (P<0.05, all). Transient transfection of BxPC-3 and SW1990 cells with PSAT1-siRNA markedly reduced the cell proliferation, migration, and invasion abilities of these cells compared to transfection with NC-siRNA (P<0.05). Knockdown of PSAT1 in pancreatic cancer cells also inhibited the expression of p-Akt and p-mTOR (P<0.05). Conclusions: The expression of PSAT1 increases in human pancreatic cancer tissues and cell lines. Additionally, PSAT1 regulates cell proliferation and in-vasion through the PI3K/Akt/mTOR pathway.

Design of intelligent platform for medical treatment quality management in perioperative period and its implementation / 中华医学图书情报杂志

After the status quo of medical treatment quality management and its relevant problems were analyzed, how to build the platform for medical treatment quality management was proposed by establishing perioperative clinical data center and intelligent management engine through data sharing, integration and reconstitution based on the present hospital information system in order to standardize perioperative medical behaviors and protect the safety of patients.

Detection of Acute Toxoplasmosis in Pigs Using Loop-Mediated Isothermal Amplification and Quantitative PCR

A loop-mediated isothermal amplification (LAMP) assay allows rapid diagnosis of Toxoplasma gondii infection. In the present study, the LAMP assay was evaluated using blood from both naturally and experimentally infected pigs. The sensitivity of the LAMP assay was compared with that of Q-PCR. Both assays detected T. gondii in the blood of experimentally infected pigs, with 100% agreement. In infected blood samples, the parasite was detected as early as 2 days post-infection and reached a peak in 3-5 days. In 216 field serum samples, the detection rates of LAMP and Q-PCR assays were 6.9% and 7.8%, respectively. This result indicates that the sensitivity of the LAMP assay was slightly lower than that of the Q-PCR assay. However, the LAMP may be an attractive diagnostic method in conditions where sophisticated and expensive equipment is unavailable. This assay could be a powerful supplement to current diagnostic methods.

Assistance with Artis Zeego Robot Imaging System in ERCP for biliary calculi / 中华消化内镜杂志

Objective To explore the value of Artis zeego robot imaging system endoscopic retrograde cholangiopancreatography (ERCP) for biliary calculi. Methods ERCP was performed on 12 patients with biliary tract dilation, diagnosed by B ultrasonography. Artis zeego robot was used simultaneously to acquire 3D images and biliary system reconstruction. The diagnostic consistency was assessed based on endoscopy and surgery if necessary. Results 3D rotating acquisition and biliary reconstruction were performed in the 12 patients. All diagnosis coincided with those of surgery and ERCP findings, achieving a consistency rate of 100%. Patients with extrahepatic bile duct stones of uncertain number (n = 2), with suspected biliary duct calculi ( n = 1 ) and with suspected intrahepatic bile duct stones ( n = 2) under ERCP were all diagnosed by the robot imaging system. Conclusion During ERCP, 3D rotating image acquisition and biliary reconstruction with Artis zeego robot is helpful for precise diagnosis of biliary tract stones.

Effects of dexamethasone on toll-like receptor(TLR)2 and 4 in CD14+ monocyte of patients during and after cardiac surgery with cardiopulmonary bypass / 中华胸心血管外科杂志

Objective To investigate the kinetics processes of TLR2 and TLR4 in CD14~+ monocyte of patients during and after cardiac surgery with cardiopulmonary bypass(CPB) and the effects of dexamethasone(DXM) on the regulation of TLR2 and 4 in CD14~+ monocyte. Methods Twenty patients undergoing elective atrial/ventricular septal defect correction were randomized to received 1 mg/kg dexamethasone or placebo before induction of anesthesia. The CD14~+ monocyte surface TLR2 and TLR4 and the intracellular HSP70 were stained and analyzed by flow cytometry, and plasma level of TNF-?, IL-6, IL-10, NO and MDA were measured at following times: before the dexamethasone or placebo were administer(T1), before starting CPB(T2), immediately after aortic declamping(T3), 30min after aortic declamping(T4), 5h after skin closure(T5) and 24h after skin closure(T6). Results Both the HSP70~+ TLR2~+ monocytes and HSP70~+-TLR4~+ monocytes,the plasma concentration of TNF-?, IL-6, NO and IL-10 were upregulated after introduction (P

Quantitative electroencephalographic analysis as a tool for predicting the depth of sedation of sedation induced by propofol, midazolam and ketamine / 中华麻醉学杂志

Objective To evaluated the acccuracy of the two quantitative electroencephalographic parameters-bispectral index (BIS) and 95% specrral edge frequency (95% SEF) for measuring the depth of sedation induced by propofol, midazolam and ketamine. Methods Forty-five ASA Ⅰ - Ⅱ patients aged 30-59 yr weighing 46-80 kg scheduled for elective general thoracic or abdominal surgery were randomized to receive an infusion of propofol at a rate of 8 mg?kg-1?h-1 (group P , n = 15) or midazolam at 0.5 mg?kg-1?h-1 (group M, n = 15) or ketamine at 4 mg?kg-1? h-1 (group K, n = 15) . The patients were unpremedicated. The depth of sedation was assessed using OAAS scale (5 = wide awake , 1 = no response to prodding or shaking ) at 3 min intervals. BIS and 95 % SEF were continuously monitored. The BIS and 95% SEF values at each OAAS score (5-1) were recorded. The relations between BIS, 95 % SEF and sedation scores were determined in each group. The ED50 values of BIS and 95% SEF50 for loss of consciousness and their 95% confidence internals were calculated. Prediction probability(Pk) values for BIS and 95% SEF were compared among the drugs. Results There were no significant differences among the 3 groups with respect to age, body weight, sex and duration of drug infusion. With increasing sedation there was a progressive decrease in BIS and 95 % SEF values in group P and M but no significant changes in BIS and 95 % SEF values were seen in group K. The BIS and 95 % SEF positively correlated with OAAS score in group P and M but not in group K. The BIS50 was 65.9 in group P and 70.7 in group M,but inestimable in group K.The 95% SEF50 was 20.4 in group P and inestimable in group M and K. The Pk values for BIS and 95 % SEF were higher in P group than in M group and were not significantly different from 0.5 indicating a very poor predictive performance . Conclusion The accuracy of BIS and 95 % SEF for assessing the depth of sedation is greater with propofol. BIS is more sensitive than 95% SEF for the same anesthesia.

The clinical evaluation of the bispectralindex of electroencephalogram in predicting movement during skin incision in patients under sevoflurane anesthesia / 中华麻醉学杂志

Objective: To evaluate the efficacy of the bispectral index (BI) in predicting patient movement on skin incision under sevoflurane anesthesia with the PK statistic. Method: Twenty-nine adult patients. scheduled for elective upper abdominal surgery, were selected. Anesthesia was induced with propofol and 4%-5% sevoflurane in oxygen and maintained with one of four randomized sevoflurane concentrations (0.6MAC, 1.0MAC. 1.5MAC and 2.0MAC) for fifteen minutes, then skin incision was performed by the surgeon at the planned site of the surgery, and each patient was observed carefully about 2 minutes to detect purposeful movement. The data of BI, 95% SEF, MAP, SABP and HR in one minute before skin incision were applied to statistically analysis. Result: The P_K was significantly greater than 0.5 for the sevoflurane level and the BI. The PK of 95% SEF. SABP and MAP were less than that of the level of sevoflu or Bl. but was significantly greater than 0.5 The P_K of HR was close to 0.5. Conclusion: These indicators of B, BP and 95% SEF may predict patient movement to skin incision under sevoflurane anesthesia. of which the BI is the most sensi tive.