Objective To investigate the biological basis of disease and syndrome by studying the spectrum of myocardial tissue metabolites in the rat model of coronary heart disease with heart blood stasis syndrome.Methods SD rats were randomly divided into sham-operation group and model group.The left anterior descending coronary artery was ligated to prepare the rat model of coronary heart disease with heart blood stasis syndrome.The general condition was observed,and the tongue chromaticity,electrocardiogram,cardiac function were detected.HE staining and transmission electron microscopy were used to observe myocardial tissue morphology and ultrastructure.UPLC-MS technology was used to investigate the differential metabolites in rat myocardial tissue,and enrichment analysis was conducted on metabolic pathways.Results Compared with the sham-operation group,the tongue chromaticity R,G,B values of model group rats were significantly reduced(P<0.05),ECG heart rate and ST segment elevation amplitude significantly increased(P<0.05),LVEF and LVFS significantly decreased,and LVIDs and LVIDd significantly increased(P<0.05).Myocardial tissue pathology revealed that the structure was blurred,inflammatory cells infiltrated,mitochondria swelled,ruptured,and dissolved,and crista structure fracture decreased.A total of 29 potential biomarkers with significant differences between the sham-operation group and the model group were identified in metabolomics(7 upregulated and 22 downregulated),with the majority of 10 pathways enriched in thiamine metabolism,arginine biosynthesis,purine metabolism,aminoacyl-tRNA biosynthesis,alanine,aspartate and glutamate metabolism,pentose and glucuronate interconversions,glycolysis/gluconeogenesis,valine,leucine and isoleucine degradation,TCA cycle,pyruvate metabolism.Conclusion Ligation of the left anterior descending coronary artery can mimic the pathological process of coronary heart disease with blood stasis syndrome in a good way,and its pathological mechanism involves the disruption of multi-level metabolic networks such as glucose metabolism,mitochondrial energy metabolism,amino acid metabolism,protein biosynthesis,and purine metabolism.

【Objective】 To identify the best surgical treatment for patients with calculous pyonephrosis by evaluating the clinical effectiveness of percutaneous nephrolithotripsy (PCNL) in stageⅠ, percutaneous nephrostomy (PCN) in stageⅠ, and percutaneous nephrolithotripsy in stageⅡ. 【Methods】 For publications published between Jan.2012 and Oct.2022, we thoroughly examined the databases of PubMed, Cochrane Library, Web of Science, EMBASE, CNKI, Wanfang, and VIP.We then chosed the literature based on the inclusion and exclusion criteria.After data were retrieved and literature quality was assessed, Review Manager software (RevMan 5.4.3, Cochrane Collaboration, Oxford, UK) was utilized to Meta-analysis. 【Results】 Out of 688 participants across 105 researches, we chosed 11 trials.Among them, 341 patients received both stageⅠPCN and stage ⅡPCNL (hereinafter referred to as phase ⅡPCNL), while 347 patients received stageⅠPCNL.According to the results of the Meta-analysis, there was no discernible difference between stage Ⅰ PCNL and stage Ⅱ PCNL in terms of stone clearance rate (P=0.95), operation duration (P=0.48), postoperative septic shock (P=0.36), or perirenal effusion infection (P=0.27).There were significant differences between stage Ⅰ PCNL and stage Ⅱ PCNL in fever (P=0.03), indwelling nephrostomy tube time (P=0.01), hospitalization costs (P=0.01), hospitalization time (P=0.01), and postoperative hospitalization time (P=0.02).The following two regimens were comparable in terms of stone clearance rate, operating time, postoperative perirenal effusion infection, and septic shock for patients with calculous pyonephrosis.Despite the fact that there were more patients who developed fever after stage ⅠPCNL, hospitalization costs were lower, indwelling nephrostomy tube time, overall hospitalization time, and postoperative hospitalization time were all reduced. 【Conclusion】 ⅠPCNL was affordable, safe, and successful for treating renal and upper ureteral calculi with pyonephrosis, and it had some promotional value in clinical practice.

The synthesis of fine chemicals using multi-enzyme cascade reactions is a recent hot research topic in the field of biocatalysis. The traditional chemical synthesis methods were replaced by constructing in vitro multi-enzyme cascades, then the green synthesis of a variety of bifunctional chemicals can be achieved. This article summarizes the construction strategies of different types of multi-enzyme cascade reactions and their characteristics. In addition, the general methods for recruiting enzymes used in cascade reactions, as well as the regeneration of coenzyme such as NAD(P)H or ATP and their application in multi-enzyme cascade reactions are summarized. Finally, we illustrate the application of multi-enzyme cascades in the synthesis of six bifunctional chemicals, including ω-amino fatty acids, alkyl lactams, α, ω-dicarboxylic acids, α, ω-diamines, α, ω-diols, and ω-amino alcohols.
Amino Acids ; Biocatalysis ; Amino Alcohols ; Coenzymes/metabolism* ; Diamines

Objective To investigate the value of interleukin-6 (IL-6) combined with Model for End-stage Liver Disease (MELD) score in predicting the prognosis of patients with hepatitis B virus (HBV)-related acute-on-chronic liver failure (HBV-ACLF). Methods A total of 86 patients with HBV-ACLF who were hospitalized in The First Affiliated Hospital of Soochow University from January 2015 to December 2018 were enrolled, and according to their survival status after follow-up for 90 days, they were divided into death group with 50 patients and survival group with 36 patients. ELISA was used to measure the serum level of IL-6, and a statistical analysis was performed for general information. The t -test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. A Pearson correlation analysis was performed for IL-6 and other variables; a binary logistic regression analysis was used to investigate the independent risk factors for the prognosis of patients with HBV-ACLF; the receiver operating characteristic (ROC) curve was used to assess the value of IL-6 combined with MELD score in predicting the prognosis of HBV-ACLF. Results There were significant differences between the death group and the survival group in hematocrit ( t =2.413), platelet count ( t =6.584), total bilirubin (TBil) ( t =-8.070), prothrombin time (PT) ( U =77.500), international standardized ratio ( U =102.000), HBV DNA ( t =-2.767), IL-6 ( t =-16.543), and MELD score ( t =-8.192), and the death group had a significantly higher level of IL-6 than the survival group (27.13±12.18 pg/mL vs 9.72±5.56 pg/mL, P < 0.001). The Pearson correlation analysis showed that IL-6 was positively correlated with TBil and PT ( r =0.579 and 0.681, both P < 0.001). The binary logistic regression analysis showed that IL-6 (odds ratio[ OR ]=1.480, 95% confidence interval [ CI ]: 1.196~1.833, P =0.007) and MELD score ( OR =1.128, 95% CI : 1.033~1.231, P < 0.001) were independent risk factors for the death of HBV-ACLF patients within 90 days. IL-6 combined with MELD score had an area under the ROC curve (AUC) of 0.891 (95% CI : 0.778~0.999), with a higher AUC than IL-6 (AUC=0.838, 95% CI : 0.687~0.989) or MELD score (AUC=0.783, 95% CI : 0.634~0.933). IL-6 combined with MELD score had a significantly higher value than IL-6 alone in predicting the prognosis of patients with HBV-ACLF ( Z =-2.257, P =0.024). Conclusion IL-6 combined with MELD score can be used as a good model for predicting the short-term prognosis of patients with HBV-ACLF.

ObjectiveTo investigate the role of coagulation function parameters and platelet indices in thrombotic events in patients with hepatitis B virus-associated acute-on-chronic liver failure (HBV-ACLF). MethodsA total of 56 patients with HBV-ACLF who were hospitalized in The First Affiliated Hospital of Soochow University from January 2015 to December 2019 were enrolled and divided into thrombotic complication (TC) group with 24 patients and non-thrombotic complication (NTC) group with 32 patients. A retrospective analysis was performed for their general clinical data on admission, and the patients were observed in terms of the changes in coagulation function, platelet count (PLT), and the platelet function-related index mean platelet volume (MPV) on days 1-7 after admission. The t-test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U rank sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. A repeated measures analysis of variance was used to compare coagulation markers within and between groups at different time points. ResultsOn admission, the TC group had a significantly younger age than the NTC group ［31.5 (29.0-34.0) years vs 48.5 (36.0-50.7) years, Z=-2.637, P=0.008］. On the day of admission, there was no significant difference in MPV between the TC group and the NTC group (P ＞0.05), while on days 2-7 after admission, there was a significant difference in MPV between the two groups (t=-2.696、-2.742、-2.894、-4.174、-3.945、-4.716,all P ＜0.01). In the TC group, MPV reached the peak value on day 5 of admission, with a mean value of 13.90±1.12 fl, which was higher than the range of normal values. On admission, all patients had a mean prothrombin time (PT) of 28.8±7.2 s, a mean activated partial thromboplastin time (APTT) of 50.5±8.7 s, and a mean international normalized ratio (INR) of 2.6±0.7, which were higher than normal values; all patients had a mean fibrinogen (Fb) level of 1.16±0.3 g/L and a mean PLT of (107.7±26.5)×109/L, which were lower than normal values. There were no significant differences in PT, APTT, Fb, INR, and PLT between the TC group and the NTC group (all P ＞0.05). ConclusionCoagulation disorder in patients with liver failure is more of a low-equilibrium state, which is complex and heterogeneous and requires individualized treatment. For patients with HBV-ACLF, the development of thrombotic events may be more associated with platelet function than PLT or conventional coagulation markers.

ObjectiveTo investigate the value of Model for End-Stage Liver Disease (MELD) score combined with platelet-to-white blood cell ratio (PWR) in predicting the short-term prognosis of patients with hepatitis B virus-associated acute-on-chronic liver failure (HBV-ACLF). MethodsA retrospective analysis was performed for the clinical data of 123 HBV-ACLF patients who were admitted to The First Affiliated Hospital of Suzhou University from June 2014 to June 2019, and according to the prognosis on day 90 after admission, these patients were divided into survival group with 53 patients and death group with 70 patients. Related clinical data were recorded, including age, sex, and total bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), serum creatinine (SCr), Albumin (Alb), prealbumin (PAB), international normalized ratio (INR), white blood cell count (WBC), lymphocyte count (LY), monocyte count (MO), neutrophil count (NE), hemoglobin (Hb), and platelet count (PLT) within 24 hours after admission, and PWR and MELD score were calculated. The t-test and the Mann-Whitney U test were used for comparison of continuous data between two groups; univariate and multivariate binary logistic regression analyses were used to analyze the association between each factor and the prognosis of HBV-ACLF; a predictive model of MELD score combined with PWR was established. The receiver operating characteristic (ROC) curve was plotted, and Youden index, cut-off value, sensitivity, and specificity were calculated; the area under the ROC curve (AUC) was calculated for MELD score alone or combined with PWR to compare their value in predicting the prognosis of HBV-ACLF patients. ResultsThere were significant differences between the two groups in TBil, ALT, SCr, INR, WBC, MO, NE, Hb, PLT, PWR, and MELD score (all P＜0.05). TBil, SCr, INR, WBC, MO, NE, and MELD score were risk factors for prognosis of HBV-ACLF patients(all P＜0.05); PWR (odds ratio ［OR］=0.883, 95% confidence interval ［CI］: 0.798-0.977, P=0.016) and MELD score (OR=1.442, 95%CI: 1.225-1698, P＜0.001) were independent predictive factors for the prognosis of HBV-ACLF patients. MELD score combined with PWR had a stronger predictive efficiency than MELD score alone in predicting the prognosis of HBV-ACLF patients ［0.895 (95%CI: 0.827-0943) vs 0.842 (95%CI: 0.765-0.902), P＜0.05］. ConclusionMELD score combined with PWR can improve the efficiency of MELD score alone in predicting the prognosis of HBV-ACLF patients.

ObjectiveTo investigate the association between mean platelet volume (MPV) and prognosis of hepatitis B virus-associated acute-on-chronic liver failure (HBV-ACLF) and the value of MPV in prognostic evaluation. MethodsA retrospective analysis was performed for the clinical data of 37 patients with hepatitis B cirrhosis, 44 patients with HBV-ACLF, and 27 patients with chronic hepatitis B, who were admitted to Department of Infectious Diseases, The First Affiliated Hospital of Soochow University, from January 2015 to December 2019, as well as 24 individuals who underwent physical examination during the same period of time. Related clinical data included sex, age, total bilirubin (TBil), creatinine (Cr), blood urea nitrogen (BUN), albumin (Alb), blood ammonia (NH3), routine blood test results, Model for End-Stage Liver Disease (MELD) score, and blood coagulation parameters. According to prognosis, the patients with HBV-ACLF patients were divided into improvement group with 19 patients and non-improvement group with 25 patients, and the patients were followed up for 3 months. The Kruskal-Wallis H test was used for comparison between multiple groups, and the Wilcoxon rank sum test was used for comparison within each group， the Mann-Whitney U test was used for comparison between two groups； a binary logistic regression analysis was used to investigate the independent influencing factors for prognosis, and the receiver operating characteristic (ROC) curve was used to predict the accuracy of variables. ResultsThere were significant differences in MPV, platelet count, prothrombin time (PT), international normalized ratio (INR), Alb, BUN, TBil, and MELD score between the liver cirrhosis group, the HBV-ACLF group, the chronic hepatitis group, and the healthy group (χ2=39.031， 75.837, 59.894, 56.033, 22.760, 83.353 and 70.299, all P＜0.001). Further comparison of MPV between two groups showed that the HBV-ACLF group had a significantly higher MPV than the healthy group and the chronic hepatitis group (Z=9.076 and 4.435, both corrected P＜0.001), and the liver cirrhosis group had a significantly higher MPV than the healthy group (Z=2.646, corrected P=0.049). For the patients with HBV-ACLF, there were significant differences in MPV, age, PT, INR, and MELD score between the improvement group and the non-improvement group (Z=-3.710， -2.726， -2.678， -2.322 and -2.610, all P＜0.05), and further binary logistic regression analysis showed that MPV (odds ratio ［OR］=1.175, 95% confidence interval ［CI］: 1.067-2.756, P=0.026) and MELD score (OR=1.643, 95% CI: 1.021-2.644, P=0.041) were independent influencing factors for the prognosis of HBV-ACLF. MPV alone, MELD score alone, and MPV combined with MELD score had an area under the ROC curve of 0.742, 0.731, and 0.791, respectively, in predicting the prognosis of HBV-ACLF, and MPV combined with MELD score had relatively high sensitivity (0.72) and specificity (0.895). ConclusionMPV is an independent influencing factor for the prognosis of HBV-ACLF and has a good value in predicting prognosis. MPV combined with MELD score has advantages over MPV or MELD score alone in predicting the prognosis of HBV-ACLF.

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.

Industrial enzymes have become the core "chip" for bio-manufacturing technology. Design and development of novel and efficient enzymes is the key to the development of industrial biotechnology. The scientific basis for the innovative design of industrial catalysts is an in-depth analysis of the structure-activity relationship between enzymes and substrates, as well as their regulatory mechanisms. With the development of bioinformatics and computational technology, the catalytic mechanism of the enzyme can be solved by various calculation methods. Subsequently, the specific regions of the structure can be rationally reconstructed to improve the catalytic performance, which will further promote the industrial application of the target enzyme. Computational simulation and rational design based on the analysis of the structure-activity relationship have become the crucial technology for the preparation of high-efficiency industrial enzymes. This review provides a brief introduction and discussion on various calculation methods and design strategies as well as future trends.
Biocatalysis ; Biotechnology ; Enzymes ; chemistry ; metabolism ; Metabolic Engineering ; Protein Engineering ; Structure-Activity Relationship

Objective: To explore the characteristics underlying Th1/Th2/Th17 expression level after entecavir (ETV) discontinuation of chronic hepatitis B (CHB) patients who were HBeAg-positive and define the role of Th1/Th2/Th17 in maintaining virological response after ETV discontinuation. Methods: We selected 112 HBeAg positive CHB patients who met the withdrawal criteria according to the guideline of prevention and treatment of chronic hepatitis B (2010 version), and we also separated them into virology sustained response (SVR) group and virological relapse (VR) group according to the recurrence in 52 weeks. We detected serum level of Th1/Th2/h17 related cytokines during 0, 12, 24 and 52 weeks follow-up to further analyze their dynamic changes and expression differences. Results: The results of the study reveals that serum levels of IFN-γ in the group of SVR were at a higher level compared with VR group during follow-up (all P<0.05). While, the serum level of IL-10 decreased in SVR group and was lower than the paired IL-10 level in VR group with significance during follow-up. In VR group, the serum level of IFN-γ decreased during the first 24 weeks after ETV withdrawal, and after then, the level rose. The serum levels of IFN-γ (Th1) in SVR patients were significantly higher than those in VR patients in the different follow-up time points after ETV discontinuation (all P<0.05). And the serum levels of IL-10 (Th2) in SVR patients were significantly lower than that in VR patients (all P<0.05). The comparison of serum levels of IL-17A (Th17) between the two groups had no significant difference (all P>0.005). Conclusions The serum level of IFN-γ in SVR group maintained a high level is conducive to maintaining virological response after ETV withdrawal and high level of IL-10 may be related to virologic relapse.