Objective To investigate the expression of autophagy marker in peripheral blood T and B lymphocytes of patients with autoimmune hepatitis (AIH) and its clinical significance. Methods Peripheral blood samples were collected from 62 AIH patients who were treated in Beijing YouAn Hospital affiliated to Capital Medical University from October 2019 to October 2020 who were treated in Beijing YouAn Hospital affiliated to Capital Medical University from October 2019 to October 2020 and 8 healthy controls to detect autophagy of T and B lymphocyte subsets, and then subgroup analyses were performed based on treatment, diagnostic type, and presence or absence of liver cirrhosis and liver failure. The t -test was used for comparison of normally distributed continuous data between two groups; the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups, and the Mann-Whitney U test was used for comparison between two groups; the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups. Results Compared with the healthy control group, the AIH group had a significantly higher mean fluorescence intensity (MFI) of the autophagy marker LC3B in CD4 + T, CD8 + T, CD19 + B, and CD4 + CD25 + T lymphocytes (all P < 0.05), especially in CD19 + B lymphocytes. The non-treatment group and the partial remission group had a significantly higher MFI of autophagy marker in CD19 + B lymphocytes than the complete remission group ( P =0.037 and 0.040); the idiopathic AIH (I-AIH) group and the drug-induced AIH(DI-AIH) group had a significantly higher MFI than the primary biliary cholangitis (PBC)-AIH overlap syndrome group ( P =0.037 and 0.031); the non-cirrhosis group and the decompensated cirrhosis group had a significantly higher MFI than the compensated cirrhosis group ( P =0.009 and 0.003); the liver failure group had a significantly higher MFI than the non-liver failure group ( P =0.042). The PBC-AIH group had a significantly higher MFI of autophagy marker in CD4 + CD25 + T lymphocytes than the I-AIH group and the DI-AIH group ( P =0.042 and 0.044), the compensated cirrhosis group had a significantly lower MFI than the non-cirrhosis group ( P =0.037), and the non-liver failure group had a significantly higher MFI than the liver failure group ( P =0.043). Conclusion AIH patients have a significant increase in the expression of autophagy marker in peripheral blood T and B lymphocyte subsets compared with healthy individuals, and the level of autophagy is associated with treatment, diagnostic type, and disease severity.

Hepatitis, Autoimmune ; Humans ; Liver Failure ; Prognosis

Objective:To evaluate the effect of deep learning based on DWI and fluid attenuated inversion recovery (FLAIR) to construct a prediction model of the onset time in acute stroke.Methods:A total of 324 cases of acute stroke with clear onset time, from January 2017 to May 2020 in Nanjing First Hospital, were retrospectively enrolled and analyzed. The patients were divided into a training set of 226 patients and a test set of 98 patients according to the complete randomization method using a 7∶3 ratio, and the patients were divided into ≤ 4.5 h and >4.5 h according to symptom onset time in each group. The acute infarction areas on DWI and the corresponding high signal area on FLAIR were manually outlined by physician. Using the InceptionV3 model as the basic model for image features extraction, the deep learning prediction model based on single sequence (DWI, FLAIR) and multi sequences (DWI+FLAIR) were established and verified. Then the area under curve (AUC), accuracy of human readings, single sequence model and multi sequence model in predicting the acute stroke onset time from imaging were compared.Results:DWI-FLAIR mismatch was found in 94 cases (94/207) of patients with symptom onset time from imaging ≤ 4.5 h, while in 28 cases (28/117) of patients with symptom onset time from imaging >4.5 h. ROC analysis showed that the AUC of DWI-FLAIR mismatch in predicting acute stroke onset time from imaging was 0.607, and the accuracy was 60.2%. The prediction model of deep learning based on single sequence showed that the AUC of FLAIR was 0.761 and the accuracy was 71.4%; the AUC of DWI was 0.836 and the accuracy was 81.6%. The AUC of predicting stroke onset time based on the multi-sequence (DWI+FLAIR) deep learning model was 0.852, which was significantly better than that of manual identification ( Z = 0.617, P = 0.002), FLAIR sequence deep learning model ( Z = 2.133, P = 0.006) and DWI sequence deep learning model ( Z = 1.846, P = 0.012). Conclusion:The deep learning model based on DWI and FLAIR is superior to human readings in predicting acute stroke onset time from imaging, which could provide guidance for intravenous thrombolytic therapy for acute stroke patients with unknown onset time.

Objective:To investigate the therapeutic effect of cardiac rehabilitation exercise on ventricular remodeling in patients with acute myocardial infarction undergoing percutaneous coronary intervention.Methods:A total of 100 patients with acute myocardial infarction undergoing percutaneous coronary intervention who received treatment in Wenzhou Central Hospital from June 2018 to June 2019 were included in this study. They were randomly divided into a rehabilitation group and a conventional treatment group ( n = 50/group). Patients in the conventional treatment group underwent conventional postoperative rehabilitation education while those in the rehabilitation group received targeted cardiac rehabilitation exercise. After surgery, all patients were followed up for 12 months. Real time three-dimensional echocardiography was used to evaluate ventricular remodeling (left ventricular ejection fraction, left ventricular end-diastolic volume , left ventricular end-systolic volume, left ventricular remodeling index) and ventricular synchrony (Tmsv-16dif, Tmsv-16sd, Tmsv16-dif%, Tmsv16-sd%) before and 3, 6 and 12 months after surgery. In addition, serum levels of ventricular remodeling indexes (fibroblast growth factor 23, PICP and PIIINP) were measured. The incidence of cardiovascular end-point events within 12 months was calculated. Results:At 3, 6 and 12 months after surgery, left ventricular ejection fraction was (51.81 ± 5.43)%, (55.88 ± 5.46)%, (55.63 ± 5.57)% in the rehabilitation group, which was significantly higher than (47.16 ± 5.38)%, (52.31 ± 5.44)%, (51.84 ± 5.59)% respectively in the conventional treatment group ( t = 4.302, 3.275, 3.396, all P < 0.05). At 3, 6 and 12 months after surgery, left ventricular end-diastolic volume was (124.65 ± 15.56) mL, (98.54 ± 14.54) mL, (99.82 ± 13.18) mL, respectively in the rehabilitation group, which was lower than (132.64 ± 16.58) mL, (112.55 ± 15.61) mL and (114.84 ± 17.35) mL, respectively in the conventional treatment group ( t = 2.485, 4.644, 4.874, all P < 0.05). At 6 and 12 months after surgery, left ventricular end-systolic volume was (52.26 ± 5.48) mL and (52.15 ± 5.32) mL respectively in the rehabilitation group, which was significantly lower than (57.92 ± 5.46) mL and (58.51 ± 5.72) mL in the conventional treatment group ( t = 5.174, 5.757, both P < 0.05). At 6 and 12 months after surgery, left ventricular remodeling index was (1.75 ± 0.42) g/mL and (1.74 ± 0.35) g/mL respectively in the rehabilitation group, which was significantly higher than (1.52 ± 0.37) g/mL and (1.50 ± 0.32) g/mL, respectively in the conventional treatment group ( t = 2.906, 3.579, both P < 0.05). At 3, 6 and 12 months after surgery, Tmsv-16dif ( t = 2.753, 4.283, 4.088, all P < 0.05), Tmsv-16sd ( t = 5.134, 4.326, 4.670, all P < 0.05), Tmsv-16dif% ( t = 7.714, 8.587, 7.800, all P < 0.05) and Tmsv16-sd% ( t = 9.004, 14.061, 10.305, all P < 0.05) respectively in the rehabilitation group, were significantly lower than those in the conventional treatment group. At 3, 6 and 12 months after surgery, fibroblast growth factor 23 ( t = 6.303, 5.053, 4.619, all P < 0.05). PICP ( t = 3.772, 2.798, 3.788, all P < 0.05) and PIIINP ( t = 3.110, 5.912, 4.294, all P < 0.05) in the rehabilitation group were significantly lower than those in the conventional treatment group. Within 12 months, the total incidence of cardiovascular end-point events in the rehabilitation group [12.00% (6/50)] was significantly lower than that in the conventional treatment [32.00% (16/50)] ( χ2 = 5.828, P < 0.05). Conclusion:Cardiac rehabilitation exercise can improve ventricular remodeling and synchrony in patients with acute myocardial infarction undergoing percutaneous coronary intervention and reduce the incidence of cardiovascular end-point events.

Heparin and heparan sulfate are a class of glycosaminoglycans for clinical anticoagulation. Heparosan N-sulfate-glucuronate 5-epimerase (C5, EC 5.1.3.17) is a critical modifying enzyme in the synthesis of heparin and heparan sulfate, and catalyzes the inversion of carboxyl group at position 5 on D-glucuronic acid (D-GlcA) of N-sulfoheparosan to form L-iduronic acid (L-IdoA). In this study, the heparin C5 epimerase gene Glce from zebrafish was expressed and molecularly modified in Escherichia coli. After comparing three expression vectors of pET-20b (+), pET-28a (+) and pCold Ⅲ, C5 activity reached the highest ((1 873.61±5.42) U/L) with the vector pCold Ⅲ. Then we fused the solution-promoting label SET2 at the N-terminal for increasing the soluble expression of C5. As a result, the soluble protein expression was increased by 50% compared with the control, and the enzyme activity reached (2 409±6.43) U/L. Based on this, site-directed mutations near the substrate binding pocket were performed through rational design, the optimal mutant (V153R) enzyme activity and specific enzyme activity were (5 804±5.63) U/L and (145.1±2.33) U/mg, respectively 2.41-fold and 2.28-fold of the original enzyme. Modification and expression optimization of heparin C5 epimerase has laid the foundation for heparin enzymatic catalytic biosynthesis.
Animals ; Carbohydrate Epimerases ; biosynthesis ; chemistry ; genetics ; Escherichia coli ; Gene Expression ; Heparin ; metabolism ; Heparitin Sulfate ; metabolism ; Iduronic Acid ; metabolism ; Zebrafish Proteins ; biosynthesis ; chemistry ; genetics

Objective:To explore the clinical characteristics and establish a corresponding prognostic scoring model in patients with early-stage clinical features of hepatitis B-induced acute-on-chronic liver failure (HBV-ACLF).Methods:Clinical characteristics of 725 cases with hepatitis B-related acute-on-chronic hepatic dysfunction (HBV-ACHD) were retrospectively analyzed using Chinese group on the study of severe hepatitis B (COSSH). The independent risk factors associated with 90-day prognosis to establish a prognostic scoring model was analyzed by multivariate Cox regression, and was validated by 500 internal and 390 external HBV-ACHD patients.Results:Among 725 cases with HBV-ACHD, 76.8% were male, 96.8% had cirrhosis base,66.5% had complications of ascites, 4.1% had coagulation failure in respect to organ failure, and 9.2% had 90-day mortality rate. Multivariate Cox regression analysis showed that TBil, WBC and ALP were the best predictors of 90-day mortality rate in HBV-ACHD patients. The established scoring model was COSS-HACHADs = 0.75 × ln(WBC) + 0.57 × ln(TBil)-0.94 × ln(ALP) +10. The area under the receiver operating characteristic curve (AUROC) of subjects was significantly higher than MELD, MELD-Na, CTP and CLIF-C ADs( P < 0.05). An analysis of 500 and 390 cases of internal random selection group and external group had similar verified results. Conclusion:HBV-ACHD patients are a group of people with decompensated cirrhosis combined with small number of organ failure, and the 90-day mortality rate is 9.2%. COSSH-ACHDs have a higher predictive effect on HBV-ACHD patients' 90-day prognosis, and thus provide evidence-based medicine for early clinical diagnosis and treatment.

Objective: To investigate the endoplasmic reticulum stress (ERS) role in the course of liver failure induced by severe hepatitis B virus (HBV) infection and its related mechanism. Methods: Liver tissue samples and clinical data [chronic hepatitis B patients (12 cases, chronic hepatitis B group), hepatic failure induced by severe hepatitis B virus (12 cases, severe hepatitis B virus liver failure group), and normal subjects (8 cases, control group)] were collected from the Beijing You'an Hospital affiliated to Capital Medical University between 2009 to 2011. Statistical analysis was performed on the clinical indicators of each group. The structure of endoplasmic reticulum in liver tissue was observed by transmission electron microscopy. Western blot and qRT-PCR were used to detect the expression of endoplasmic reticulum stress and apoptosis-related factors, including glucose-regulated protein (Grp), and C/EBP homologous protein (CHOP). Frozen sections of liver tissues were prepared for immunofluorescence test. All data were expressed as mean ± standard deviation. LSD-t test was used to compare the results between groups. A p value < 0.05 was considered as statistically significant. Results: Transmission electron microscopy showed that the morphological structure of the endoplasmic reticulum was damaged in both groups (chronic hepatitis B and liver failure induced by severe hepatitis B virus), and liver failure induced by severe hepatitis B virus group was more critical. Western blot and qRT-PCR showed that Grp78, Grp94 and Caspase-4 were highly expressed in normal group and chronic hepatitis B group, and the relative protein expressions were 1.20 ± 0.13 and 0.78 ± 0.11, 0.90 ± 0.06 and 0.11 ± 0.01, 0.15 ± 0.02 and 0.22 ± 0.04, respectively. The expression of protein was weakened in liver failure induced by severe hepatitis B virus group (relative protein expression was 0.01 ± 0, 0.01 ± 0, and 0.11 ± 0.02, respectively).There was a statistically significant difference between the two groups (P < 0.05). The expression of CHOP was consistent with the results of immunofluorescence, and increased with the stressing of injury. Conclusion During the course of severe hepatitis B infection, dysregulated endoplasmic reticulum stress activated mild stress in chronic hepatitis B group, while severe stress in hepatic failure induced by severe hepatitis B virus group. Therefore, endoplasmic reticulum stress plays an important and complex role in the pathogenesis of hepatic failure induced by severe hepatitis B virus.

Data of patients with maxillofacial and jugular venous malformations involving isthmus faucium area from January 2012 to July 2016 were collected. Patients were questioned about the medical history before operation, and the airway was adequately evaluated. The patients diagnosed as having non-difficult airways were endotracheally tubed using fast induction of anesthesia. Tracheal intubation was per-formed using fast induction of anesthesia combined with topical anesthesia after visual laryngoscopy in the patients assessed as having difficult airways. Endotracheal intubation was guided with a visual hard endo-scope or a fibrobronchoscope in the patients with difficulty in opening mouth after multiple treatments. Post-operative airway management was as follows: the tracheal tube was removed after extubation, the tracheal tube was retained for 24-48 h, or preventive tracheotomy was performed. Oxygen was inhaled by mask. A total of 157 patients were included in this study, 55 patients diagnosed as having difficult airways, and a-mong the 55 patients, 87％ cases were intubated after visual laryngoscopy and 13％ cases received tracheot-omy. There were 10 patients with difficulty in opening mouth after multiple treatments and 5 cases under-went tracheotomy in the outer hospital. Sixteen patients presented with a transient increase in airway pres-sure during intraoperative injection of anhydrous ethanol. There were 106 cases in whom the tracheal tube was removed immediately after emergence, and among them, 32. 1％ cases needed tongue traction and 2. 8％ cases underwent emergency tracheotomy after extubation. Thirty-eight patients needed to retain the tracheal tube for 24-48 h after operation, and among them, 37％ cases needed tongue traction and 3％ ca-ses required emergency tracheotomy after extubation. Thirteen cases underwent preventive tracheotomy. The preoperative visit and assessment are especially important, appropriate airway management strategies should be developed, vital signs should be closely observed during operation, and the timing of extubation should be grasped for this type of patients, and the SpO2 and airway pressure should be mainly observed during op-eration especially for the patients who underwent anhydrous ethanol injection.

In industrial fermentation processes, bacteria have to adapt environmental stresses. Sometimes, such a self-adaption does not work and will cause fermentation failures, although such adaptation also can generate unexpected positive effects with improved fermentation performance. Our review introduces cell self-adaption to environmental variations or stress, process optimization based on such self-adaptions, with heterologous proteins production by Pichia pastoris and butanol fermentation as examples. Our review can sever as reference for fermentation optimization based on cell self-adaption.
Adaptation, Physiological ; Butanols ; metabolism ; Environment ; Fermentation ; Pichia ; cytology ; metabolism

Objective To investigate the endoplasmic reticulum stress(ERS)role in the course of liver failure induced by severe hepatitis B virus(HBV)infection and its related mechanism.Methods Liver tissue samples and clinical data [chronic hepatitis B patients(12 cases,chronic hepatitis B group),hepatic failure induced by severe hepatitis B virus(12 cases,severe hepatitis B virus liver failure group),and normal subjects(8 cases,control group)] were collected from the Beijing You'an Hospital affiliated to Capital Medical University between 2009 to 2011.Statistical analysis was performed on the clinical indicators of each group.The structure of endoplasmic reticulum in liver tissue was observed by transmission electron microscopy.Western blot and qRT-PCR were used to detect the expression of endoplasmic reticulum stress and apoptosis-related factors,including glucose-regulated protein(Grp),and C/EBP homologous protein(CHOP).Frozen sections of liver tissues were prepared for immunofluorescence test.All data were expressed as mean±standard deviation.LSD-t test was used to compare the results between groups.A p value<0.05 was considered as statistically significant.Results Transmission electron microscopy showed that the morphological structure of the endoplasmic reticulum was damaged in both groups(chronic hepatitis B and liver failure induced by severe hepatitis B virus),and liver failure induced by severe hepatitis B virus group was more critical.Western blot and qRT-PCR showed that Grp78,Grp94 and Caspase-4 were highly expressed in normal group and chronic hepatitis B group,and the relative protein expressions were 1.20±0.13 and 0.78±0.11,0.90±0.06 and 0.11±0.01,0.15±0.02 and 0.22±0.04,respectively.The expression of protein was weakened in liver failure induced by severe hepatitis B virus group(relative protein expression was 0.01±0,0.01±0,and 0.11±0.02,respectively).There was a statistically significant difference between the two groups(P<0.05).The expression of CHOP was consistent with the results of immunofluorescence,and increased with the stressing of injury.Conclusion During the course of severe hepatitis B infection,dysregulated endoplasmic reticulum stress activated mild stress in chronic hepatitis B group,while severe stress in hepatic failure induced by severe hepatitis B virus group.Therefore,endoplasmic reticulum stress plays an important and complex role in the pathogenesis of hepatic failure induced by severe hepatitis B virus.