Inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ) is one of important kinases in inflammation to phosphorylate inhibitor of nuclear factor kappa-B (IκBα) and then activate nuclear factor kappa-B (NF-κB). Inhibition of IKKβ has been a therapeutic strategy for inflammatory and autoimmune diseases. Here we report that IKKβ is constitutively activated in healthy donors and healthy Ikkβ ^C46A (cysteine 46 mutated to alanine) knock-in mice although they possess intensive IKKβ-IκBα-NF-κB signaling activation. These indicate that IKKβ activation probably plays homeostatic role instead of causing inflammation. Compared to Ikkβ ^WT littermates, lipopolysaccharides (LPS) could induce high mortality rate in Ikkβ ^C46A mice which is correlated to breaking the homeostasis by intensively activating p-IκBα-NF-κB signaling and inhibiting phosphorylation of 5' adenosine monophosphate-activated protein kinase (p-AMPK) expression. We then demonstrated that IKKβ kinase domain (KD) phosphorylates AMPKα1 via interacting with residues Thr183, Ser184, and Thr388, while IKKβ helix-loop-helix motifs is essential to phosphorylate IκBα according to the previous reports. Kinase assay further demonstrated that IKKβ simultaneously catalyzes phosphorylation of AMPK and IκBα to mediate homeostasis. Accordingly, activation of AMPK rather than inhibition of IKKβ could substantially rescue LPS-induced mortality in Ikkβ ^C46A mice by rebuilding the homeostasis. We conclude that IKKβ activates AMPK to restrict inflammation and IKKβ mediates homeostatic function in inflammation via competitively phosphorylating AMPK and IκBα.

ObjectiveTo investigate the effect of betulinic acid （BA） on apoptosis and autophagy of human colorectal cancer SW620 cells and the regulatory role of phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. MethodCell viability was detected by methyl thiazolyl tetrazolium （MTT） colorimetry to determine the optimal administration time and dosage for subsequent experiments. Four groups were designed， including blank group and low-， medium-， and high-dose BA groups. Hematoxylin-eosin （HE） staining was conducted for the observation of SW620 cell morphology， and annexin-V/propidium iodide double staining for the determination of apoptosis rate in SW620 cells. Hoechst33258 staining and MDC staining were used for the observation of apoptosis and autophagy， respectively. Western blotting was employed to determine the protein levels of B-cell lymphoma/leukemia-2（Bcl-2）-associated X protein （Bax）， aspartate proteolytic enzyme-9 （Caspase-9）， activated aspartate proteolytic enzyme-3 （cleaved Caspase-3）， microtubule-associated protein 1 light chain 3 （LC3）， the mammalian homolog of yeast Atg6 （Beclin-1）， p62， phosphorylated PI3K （p-PI3K）， phosphorylated Akt （p-Akt）， and phosphorylated mTOR （p-mTOR） in SW620 cells. ResultBA inhibited the activity of SW620， HT29， and HCT116 cells in a concentration- and time-dependent manner. The cells treated with BA for 48 h had lower viability than those treated for 24 h （P<0.05， P<0.01）. The half maximal inhibitory concentration (IC₅₀） value of BA at the time point of 48 h was also lower than that at the time point of 24 h （P<0.01）， and that for SW620 cells was the minimum. BA induced the apoptosis in a concentration-dependent manner and increased the autophagosomes. Compared with the blank group， BA increased the apoptosis rate （P<0.01）， up-regulated the protein levels of Bax， Caspase-9， cleaved Caspase-3， and LC3 Ⅱ （P<0.05， P<0.01）， and down-regulated the protein levels of p62， p-Akt， p-PI3K， and p-mTOR （P<0.01）. Additionally， medium- and high-dose BA up-regulated the protein level of beclin-1 （P<0.01）. ConclusionBA may inhibit the activity of SW620 cells by hindering the PI3K/Akt/mTOR signaling pathway to induce cell apoptosis and autophagy.

Objective: To investigate the safety and feasibility of caudal-medial approach combined with "page-turning" middle lymphadenectomy in the laparoscopic right hemicolectomy. Methods: A descriptive cohort study was conducted. Clinical data of 35 patients who underwent laparoscopic radical right hemicolectomy using caudal-medial approach combined with "page-turning" middle lymphadenectomy at Department of Gastrointestinal Surgery, Guangdong Hospital of Chinese Medicine from April 2018 to May 2020 were retrospectively analyzed. All operations were performed consecutively by the same surgeon. The caudal-medial approach was used to dissect the right Toldt's fascia and the anterior pancreaticoduodenal space in a caudal-to-cranial and medial-to-lateral manner guided by the duodenum. The "page-turning" middle lymphadenectomy was used to dissect the mesocolon along the superior mesenteric vein with ileocolic vein, Henle's trunk and pancreas exposed preferentially. Results: All the 35 patients completed the operation successfully, and there was no damage and bleeding of superior mesenteric vessels and their branches. The operative time was (186.9±46.2) minutes, and the blood loss was 50 (10-200) ml. The first time to flatus was (2.1±0.6) days, and the time to fluid intake was (2.5±0.8) days. The postoperative hospital stay was 6 (3-18) d. The overall morbidity of postoperative complication was 8.6% (3/35), including grade II in 1 cases (2.8%) and grade IIIa in 2 case (5.7%) according to the Clavien-Dindo grading standard. The total number of lymph node dissected was 30.2±5.6, and the positive lymph node was 0 (0-7). Tumor staging revealed 5 cases of stage I, 18 cases of stage II, 11 cases of stage III, and 1 case of stage IVA. In this study, the median follow-up time was 15 (4-29) months. One patient died due to cerebrovascular accident 12 months after surgery, and no tumor recurrence or metastasis was observed in all other patients. Conclusions: Laparoscopic radical right hemicolectomy using caudal-medial approach combined with "page-turning" middle lymphadenectomy is safe and feasible. The anterior pancreaticoduodenal space is preferentially mobilized, which reduces the difficulty of central vascular dissection.
Cohort Studies ; Colectomy ; Colonic Neoplasms/surgery* ; Humans ; Laparoscopy ; Lymph Node Excision ; Retrospective Studies

Objective:To investigate the risk factors for death in children with acute necrotizing encephalopathy (ANE) in pediatric intensive care unit (PICU).Methods:This was a multicenter retrospective study. Thirty-nine children with ANE were from PICUs in 4 centers from December 1, 2014 to December 1, 2020. The 4 participating centers were Beijing Children′s Hospital, Shengjing Hospital of China Medical University, Hebei Children′s Hospital, and Bao′an Maternity & Child Health Hospital. Patients were divided into survival and non-survival groups by the outcome at discharge, and the differences in clinical data between the two groups were compared. Risk factors for death in children with ANE and the odds ratios ( OR) were analyzed by univariable Logistic regression. Results:Thirty-nine children with ANE were included. There were 18 males and 21 females. The median onset age was 30 months. The mortality at discharge was 41% (16/39). The onset age of most patients (74%, 29/39) was younger than 4 years old. Influenza virus was the most common precursor infection (80%, 20/25). Patients with shock at PICU admission were more common in the non-survival group (12/16 vs. 17% (4/23), P=0.001). Glasgow coma score (GCS) at PICU admission was significantly lower in the non-survival group than survival group (3 (3, 6) vs. 6 (5, 7), Z=-2.598, P=0.009). The optimal cut-off value was 4. The proportion of patients with GCS ≤ 4 at PICU admission was higher in the non-survival group (10/16 vs. 22% (5/23), P=0.018). ANE severity score (ANE-SS) at PICU admission was significantly higher in the non-survival group (5 (2, 6) vs. 2 (1, 4), Z=-2.436, P=0.015). The proportion of patients with high risk ANE-SS was higher in non-survival group than the survival group (9/16 vs. 22% (5/23), P=0.043). The proportion of application of high-dose methylprednisolone (20 mg/(kg·d)) was significantly higher in survival group than non-survival group (43% (10/23) vs. 1/13, P=0.031). Univariable Logistic regression indicated that risk factors for death in children with ANE were shock ( OR=14.250, 95% CI 2.985-68.018, P=0.001), GCS≤4 ( OR=6.000, 95% CI 1.456-24.733, P=0.013) and high risk ANE-SS ( OR=4.629, 95% CI 1.142-18.752, P=0.032) at PICU admission. Conclusions:ANE usually occurs in children under 4 years old after influenza infection. Shock, GCS≤4 and high risk ANE-SS at PICU admission were risk factors for death in children with ANE. High-dose methylprednisolone may improve the prognosis of children with ANE.

Objective:To evaluate the clinical efficacy and safety of endoscopic submucosal dissection (ESD) for treatment of duodenal lesions.Methods:A retrospective analysis was performed on the data of 45 patients with 46 duodenal lesions who underwent ESD at the Second Xiangya Hospital of Central South University from January 2011 to May 2019. The lesion features, en bloc resection rate, complete resection rate, complications, postoperative pathology and recurrence were assessed.Results:Among the 45 patients, 20 were males and 25 were females, with age of 52.0±11.8 years. Of the 46 lesions, 31 (67.4%) were located in the duodenal bulb, 12 (26.1%) in the descending part, and 3 (6.5%) at the junction of bulb and descending part. The diameter of the lesions was 2.4±1.9 cm. There were 14 (30.4%) lesions originated from mucosal layer, 29 (63.1%) from submucosal layer, and 3 (6.5%) from muscularis propria.Postoperative pathology showed 11 (23.9%) cases of Brunner gland tumors, 9 (19.6%) neuroendocrine tumors, 5 (10.9%) ectopic pancreas, 5 (10.9%) lipomas, and 16 (34.8%) other pathological patterns. All 45 patients with 46 lesions underwent ESD successfully, and the en bloc resection rate was 100.0% (46/46), complete resection rate was 91.3% (42/46). Intraoperative bleeding occurred in 1 case (2.2%), which was successfully treated by endoscopy. One (2.2%) delayed perforation occurred and was treated by surgical intervention. Electrocoagulation syndrome occurred in 1 case (2.2%), which was relieved after conservative medical therapy. Two cases received further surgery after ESD. The mean hospital stay was 6.2 days (ranged 2-21 days) and no death occurred. Forty-one cases were followed up for 1-78 months, with mean time of 30 months. During the follow-up period, local recurrence occurred in 1 patient (2.4%).Conclusion:ESD is an effective and safe treatment for duodenal lesions and has a good clinical practical value.

Objective: To investigate the effect of intracoronary administration of nicorandil prior to primary percutaneous coronary intervention (PPCI) on myocardial perfusion and short-term clinical outcomes in patients with ST-segment elevation myocardial infarction (STEMI). Methods: A total of 158 patients with STEMI undergoing PPCI from January 2014 to December 2015 in Fuzhou General Hospital were enrolled consecutively in this prospective controlled randomized trial. Patients were assigned into three groups with random number table: the nicorandil group (patients received intracoronary administration of 6 mg nicorandil after guide wire or balloon successfully crossed the target lesion, n=53), the nitroglycerin group (patients received intracoronary administration of 300 μg nitroglycerin after after guide wire or balloon successfully crossed the target lesion, n=52) and the control group(patients received routine treatment, n=53). The primary outcomes were myocardial perfusion, including the levels of corrected TIMI frame count (cTFC), and the incidence of no reflow or slow flow after PPCI. The secondary outcomes included the incidence of major adverse cardiovascular events (MACE) during hospitalization (all-cause death, reperfusion arrhythmia within 2 hours after PPCI, angina within 24 hours after PPCI, new heart failure or worsening cardiac function, and repeat revascularization) and within 3 months of follow-up (all-cause death, nonfatal myocardial infarction, repeat revascularization, post-infarction angina, and re-hospitalization for congestive heart failure). Results: The age of enrolled patients was (62.9±11.3) years old, and 130 cases (82.3%) of them were male. The median time of symptom-onset to balloon was 4.50 (3.20, 6.43) hours. There were significantly difference in cTFC immediately after PPCI((21.68±7.43)frames, (24.74±8.66)frames, and(27.06±10.40)frames), incidence of no reflow or slow flow after PPCI(5.7%(3/53), 13.5%(7/52), and 22.6%(12/53)), ST-segment resolution at 2 hours after procedure(90.6%(48/53), 84.6%(44/52), and 74.5%(38/53)), and reperfusion arrhythmia at 2 hours after procedure(15.1%(8/53), 36.6%(19/52), and 34.0%(18/53)) among the 3 groups(P<0.01 or 0.05). In the multivariate logistic regression models, intracoronary administration of nicorandil could lower the cTFC level (OR=0.17, 95%CI 0.10-0.41, P=0.001), acted as a protecting factor on lowering the incidence of no reflow or slow flow (OR=0.13, 95%CI 0.02-0.96, P=0.045) and reperfusion arrhythmia (OR=0.26, 95%CI 0.09-0.74, P=0.012), as well as facilitating the ST-segment resolution at 2 hours after procedure (OR=4.62, 95%CI 1.14-18.82, P=0.033). However, observed parameters were similar between intracoronary administration of nitroglycerin group compared with control group (all P>0.05). MACE within 3 months of follow-up were similar among the 3 groups(all P>0.05). Conclusion Intracoronary administration of nicorandil prior to balloon dilation can significantly improve the myocardial perfusion and reduce the occurrence of reperfusion arrhythmia during PPCI for STEMI, but does not affect the short-term prognosis in STEMI patients.

Objective To analyze the medical service pricing policy of 30 provinces and autonomous regions in China and recommend on problems and challenges, for reference of medical service pricing reform in the future.Methods Literature review and policy analysis were used for summary and performance appraisal of medical service pricing of China.Results Most of the areas surveyed followed the step-by-step approach to gradually adjust pricing, while a few areas completed it in a one-time adjustment manner.2015 witnessed most areas, up to 20 (66.67%), in price adjustment;their price adjustments focused on price reduction of large-scale medical equipment inspection fees and some medical tests.On the other hand, prices of diagnosis, nursing care, and treatment which are labor-intensive were increased.Conclusions Specifications of medical services are improving, yet there exist great gaps in the convergence of medical insurance.It is advisable to expedite the convergence of the new version specifications, to achieve value regression and dynamic price adjustment of medical services.

Objective To investigate the accuracy and feasibility of brachial artery peak velocity variation (ΔVpeakbrach) and inferior vena cava variability (VIVC) as indicators of fluid responsiveness in critically ill patients. Methods A single-center prospective observation was conducted. The patients on mechanical ventilation with spontaneously breathing admitted to Department of Critical Care Medicine of the Second Affiliated Hospital of Kunming Medical University from June 2013 to August 2015 were enrolled. The patients were diagnosed as severe sepsis or sepsis shock. The peak velocity in brachial artery and diameter of the inferior vena cava at the end of inspiration and expiration was measured by bedside portable ultrasonic machine, and then ΔVpeakbrach and VIVC were calculated. The hemodynamic parameters were collected at baseline and after volume expansion (VE). The stroke volume (SV) was measured by pulse-indicated continuous cardiac output (PiCCO). Patients were classified as responders or non-responders according to the variation of SV (ΔSV) increased ≥ 15% or not after VE. Receiver operating characteristic curve (ROC) was plotted to evaluate the sensitivity and specificity of ΔVpeakbrach and VIVC in predicting volume responsiveness. Results Among 58 patients after VE, 32 patients were defined as responders and the rest 26 were defined as non-responders.There were no differences in gender, age, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, dose of vasoactive agent, ventilator parameters and infection site. Compared with baseline hemodynamic parameters, heart rate (HR) was decreased (bpm: 95±18 vs. 103±21), and systolic blood pressure (SBP) was increased [mmHg (1 mmHg = 0.133 kPa): 92±8 vs. 80±7] after VE in responders; central venous pressure (CVP) was increased after VE in non-responders (mmHg: 11±4 vs. 8±3, all P < 0.05). The ΔVpeakbrach [(15.4±4.3)% vs. (11.2±3.5)%] and VIVC [(18.6±4.1)% vs. (14.3±3.6)%] in responders were significantly increased as compared with those of non-responders (both P < 0.05). The area under ROC curve (AUC) of ΔVpeakbrach for predicting volume responsiveness was 0.816. When the cut-off value of ΔVpeakbrach was ≥ 13.3%, the sensitivity was 71.9%, and the specificity was 80.8%. AUC of VIVC for predicting volume responsiveness was 0.733. When the cut-off value of VIVC was ≥ 19.25%, the sensitivity was 53.1%, and the specificity was 88.5%. Conclusion ΔVpeakbrach and VIVC are reliable indicators for predicting volume responsiveness in critical patients.

Objective To provide useful information for the diagnosis and differentiation through exploring the tumor with polyserositis as the initial symptom.Methods There was a case present with fever,poor spirit,abdominal distention and edema in Children's Hospital of Hebei Province.The characteristics and results of physical examination were summarized and discussed according to the condition evolution phases.The possible etiological factors were analyzed.The treatment was adjusted.Further physical examination was improved.The final diagnosis was tracked.Results After getting admitted to hospital,the patient received CT scan and ultrasound examination for many times.The results suggested the pericardial effusion,pleural effusion,peritoneal effusion and cardiac insufficiency.The child had made a slight improvement through treatment.However,he had hemiparesis.Cerebral infarction was demonstrated through magnetic resonance imaging(MRI),magnetic resonance angiography (MRA),magnetic resonance venography (MRV).Ultrasound showed that there was more pericardial effusion,aortic root solid low echo mass,pulmonary arterial wall stiffness,left and right pulmonary artery blood flow speed increased.Neck and chest enhanced CT showed mediastinal lesions.Pathology examination results suggested myeloid sarcoma after referral to superior hospital.Conclusions When children had an unexplained polyserositis,more comprehensive analysis were needed.The illness condition should be explained with Monism as far as possible.Repeated inspections would be necessary when the pathogenesis was not clear.Careful watch for the tumor should be kept.

Halohydrin dehalogenase is of great significance for biodegradation of the chlorinated pollutants, and also serves as an important biocatalyst in the synthesis of chiral pharmaceutical intermediates. A putative halohydrin dehalogenase (HheTM) gene from Tistrella mobilis KA081020-065 was cloned and over-expressed in Escherichia coli BL21 (DE3). The recombinant enzyme was purified by Ni-NTA column and characterized. Gel filtration and SDS-PAGE analysis showed that the native form of HheTM was a tetramer. It exhibited the highest activity at 50 degrees C. The nature and pH of the buffer had a great effect on its activity. The enzyme maintained high stability under the alkaline conditions and below 30 degrees C. HheTM catalyzed the transformation of ethyl(S)-4-chloro-3-hydroxybutyrate in the presence of cyanide, to give ethyl (R)-4-cyano-3-hydroxybutyrate, a key intermediate for the synthesis of atorvastatin.
3-Hydroxybutyric Acid ; chemistry ; Bacterial Proteins ; genetics ; metabolism ; Cloning, Molecular ; Escherichia coli ; Hydrolases ; genetics ; metabolism ; Hydroxybutyrates ; chemistry ; Recombinant Proteins ; genetics ; metabolism ; Rhodospirillaceae ; enzymology ; genetics