Celastrol and wilforlide A are the main active triterpenoids of the traditional Chinese medicine Lei Gong Teng, which have anti-tumour, anti-inflammatory and immunosuppressive activities, and are the material basis for the clinical efficacy of Lei Gong Teng-related Chinese medicinal preparations. By analysing the biosynthetic pathway of active ingredients, optimizing genetic elements and utilizing "cell factory" to produce triterpenoids heterologously will be an effective way to obtain from Tripterygium wilfordii in the future in a "low-cost and high-efficient" manner. CYP712Ks are the first cytochrome P450s involved in the skeleton modification of friedelane-type and oleanane-type triterpenoids in T. wilfordii, and they can catalyse the generation of polpunonic acid from friedelin and 3-epi-katonic acid from β-amyrin by carboxylation at C-29 position. In this study, four multifunctional TwCYP712K1/2/3/5 were used to clarify the catalytic function and substrate selection preference using in vivo functional characterization in Saccharomyces cerevisiae. The spatial structure of the protein-substrate binding was clarified through homology modeling and molecular docking, and then the differential amino acids in the active pocket of the protein were mutated to clarify the crucial amino acids determining the catalytic function and the selection of substrate structure. A total of 63 mutant elements were constructed, and the amino acid sites affecting the carboxylation function of TwCYP712Ks were analyzed. In particular, the key amino acids affecting the substrate selectivity of TwCYP712K2 towards oleanane-type and friedelane-type triterpenoids were revealed and the TwCYP712K2^F127I and TwCYP712K2^A227T mutants would result in a reversal of the product ratio. In conclusion, four proteins of TwCYP712Ks were semi-rationally designed by homologous protein alignment and mutual mutation to elucidate multiple amino acid sites determining the catalytic function of the proteins, and a series of activity-enhancing or altering mutants were obtained, which provide abundant catalytic elements for the biosynthesis of active triterpenoids from T. wilfordii, and the mechanism of carboxylation in the C-29 position was initially elucidated.

Objective To compare the influence between self-monitoring of blood gluocose(SMBG)combined with digital diabetes management and traditional management mode on the related clinical indexes in the patients with type 2 diabetes mellitus(T2DM).Methods A total of 100 patients with T2DM treated in the endocrinology and metabolism outpatient department of this hospital from January 2022 to June 2022 and meeting the inclusion criteria of this study were successively included.They were divided into the experimental group and control group.The experimental group was managed by SMBG combined with digital diabetes man-agement mode,while the control group adopted the traditional management mode,the outpatient clinic follow up once a month.After 6 months of follow-up,fasting blood glucose,glycosylated hemoglobin(HbA1c),low density lipoprotein cholesterol(LDL-C)and urinary microalbumin/creatinine ratio(UACR)were compared between the two groups.Results The FBG,HbA1c,LDL-C,and UACR of the experimental group decreased after intervention when compared with baseline.Compared with the control group,the FBG[8.7(7.7,9.2)mmol/L vs.10.8(8.8,12.7)mmol/L,Z=-4.660,P＜0.001],HbA1c[6.3％(5.3,7.8)％vs.8.5％(7.2,10.0)％,Z=-5.130,P＜0.001],LDL-C[2.6(1.8,3.1)mmol/L vs.3.3(2.6,4.0)mmol/L,Z=-4.112,P＜0.001],UACR[16.1(3.5,46.5)mg/g vs.58.4(11.9,108.0)mg/g,Z=-2.220,P=0.026]for patients in the expriemental group after intervention were significantly decreased.Conclusion SMBG combined with digital diabetes management model can significantly improve the clinical indicators of patients.

Objective To explore the efficacy and safety of ticagrelor de-escalation and nicorandil therapy in elderly patients with acute coronary syndrome(ACS)after percutaneous coronary intervention(PCI).Methods A total of 300 elderly patients with ACS were selected from the Sixth and Seventh Medical Center of Chinese PLA General Hospital and Beijing Chaoyang Integrative Medicine Emergency Rescue and First Aid Hospital from November 2016 to June 2019,including 153 males and 147 females,aged>65 years old.All the patients received PCI,and all had double antiplatelet therapy(DAPT)scores≥2 and a new DAPT(PRECISE-DAPT)score of≥25.All patients were divided into two groups by random number table method before operation:ticagrelor group(n=146,ticagrelor 180 mg load dose followed by PCI,and ticagrelor 90 mg bid after surgery)and ticagrelor de-escalation + nicorandil group(n=154,ticagrelor 180 mg load dose followed by PCI,ticagrelor 90 mg bid+nicorandil 5 mg tid after surgery,changed to ticagrelor 60 mg bid+ nicorandil 5 mg tid 6 months later).Follow-up was 12 months.The composite end points of cardiovascular death,myocardial infarction and stroke,the composite end points of mild hemorrhage,minor hemorrhage,other major hemorrhage and major fatal/life-threatening hemorrhage as defined by the PLATO study,and the composite end points of cardiovascular death,myocardial infarction,stroke and bleeding within 12 months in the two groups were observed.Results The comparison of general baseline data between the two groups showed no statistically significant difference(P>0.05).There was also no significant difference in the composite end points of cardiovascular death,myocardial infarction and stroke between the two groups(P>0.05).The cumulative incidence of bleeding events in ticagrelor de-escalation + nicorandil group was significantly lower than that in ticagrelor group(P<0.05),while the composite end points of cardiovascular death,myocardial infarction,stroke and bleeding were also significantly lower than those in tecagrelor group(P<0.05).Conclusion In elderly patients with ACS,the treatment of ticagrelor de-escalation + nicorandil after PCI may not increase the incidence of ischemic events such as cardiovascular death,myocardial infarction or stroke,and it may reduce the incidence of hemorrhagic events.

Objective:To establish the animal model of cervicofacial venous malformations(VMs)by surgical reconstruction of exter-nal jugular vein in sheep.Methods:The external jugular veins of 5 sheep were dissected,and the position,course,branch and exter-nal diameter were observed and measured.The models of VMs with draining and returning veins were constructed by suturing or constric-ting the proximal part of main trunk and ligating or constricting the distal part of the jugular or branch veins.The animal model was eval-uated by Doppler ultrasound,gross observation and histological observation at the 4th week after surgery.Results:The external jugular veins of sheep is in the lateral side of bilateral neck,and the main trunk is formed by the maxillary vein and lingual facial vein.The ex-ternal diameter ranges from 6 to 12 mm,with an average external diameter of 9.3 mm.Immediately after the external jugular vein was sutured and narrowed at the proximal part of the main vein,the distal part of the vein branch was ligated or narrowed,the blood flow speed slowed down and the veins in the model area bulged.4 weeks after surgery,gross observation showed that most veins narrowed and thrombosis was formed in part of the venous lumen.The central region of some specimens was dilated,and the peripheral collateral veins were dilated in some models.Doppler ultrasonography showed that the lumens of most veins were dilated and the returning veins and the inflow veins were narrowed.Colored blood flow was seen in the lumen.Histological observation showed that the structure of vein endothelium and wall was close to the normal vein,and the vein vessel wall of some specimens was thickened.Conclusion:The VMs model estab-lished by external jugular vein of sheep basically meets the re-quirements and is expected to be used in the therapeutic meth-odology research of cervicofacial VMs.

Objective To explore the value of ultrasonic quantitative measurement of hepatorenal index(HRI)for diagnosing non-alcoholic fatty liver disease(NAFLD)in children.Methods Abdominal ultrasound and upper abdominal MRI data of 70 obese children were retrospectively analyzed.ROI with different sizes and shapes of liver and right kidney were delineated on longitudinal and transverse ultrasound images,respectively,and the echo intensity of ROIs were measured to obtain HRIsmall ROI on longitudinal section,HRIsmall ROI on transverse section,HRIlarge ROI on longitudinal section and HRIlarge ROI on transverse section,i.e.HRI1,HRI2,HRI3,HRI4,while the gray,skewness and kurtosis of liver ultrasound image were recorded.Liver proton density fat fraction(PDFF)were measured based on MRI,and NAFLD was diagnoses taken PDFF≥6%as standard.The correlations of HRI with PDFF and liver ultrasound image related parameters were analyzed.Taken MRI as the standard,receiver operating characteristic(ROC)curve was drawn to evaluate the diagnostic efficacy of HRI for NAFLD.Multivariate logistic regression analysis was performed taken age,sex,body mass index(BMI)percentile,HRI3 and liver ultrasound image related parameters as independent variables and MRI diagnosis of NAFLD as dependent variable to screen the predictors of MRI diagnosis of NAFLD.Results HRI1,HRI2,HRI3 and HRI4 obtained with ultrasound was 1.89±0.52,1.88±0.55,1.97±0.51 and 1.92±0.55,respectively.PDFF obtained with MRI was(12.53±3.14)%,and diagnosed NAFLD in 34 cases.HRI and PDFF had moderate positive correlation(r=0.51-0.61,all P＜0.01).The correlation between HRI3 and PDFF was the strongest(r=0.61),and HRI3 was weakly correlated with liver gray3(r=-0.270,P=0.020),with area under the curve(AUC)for diagnosing NAFLD of 0.93(P＜0.01).BMI percentile(OR=1.06),HRI3(OR=34.20)and liver gray3(OR=0.79)were all predictive factors for MRI diagnosis of NAFLD.Conclusion Ultrasonic quantitative measurement of HRI had high clinical value for diagnosing NAFLD in children.

Objective To investigate the clinical characteristics and risk factors of the occurrence of hypoxic hepatitis(HH)in children with shock.Methods Clinical data(general situation,clinical situation and prognosis)of 234 children with different types of shock admitted to the pediatrics department of Hainan Women and Children's Medical Center from January 2016 to December 2023 were collected.Serum biochemical indexes on day 1(d1),day 2(d2),day 3(d3),day 5(d5),day 7(d7)of children with shock were collected,including serum alanine aminotransferase(ALT),glutamate aminotransferase(AST),lactate dehydrogenase(LDH),alkaline phosphatase(ALP),albumin(ALB),total bilirubin(TBIL),urea nitrogen(BUN),C reactive protein(CRP),international standardized time(INR)and lactate.And the clinical characteristics and risk factors of the occurrence of HH in children with shock were analyzed.Results A total of 234 cases(150 cases of males)of different types of shock in children were enrolled,ages ranged from 1 month to 16 years old.According to whether developed HH,the children with shock were divided into HH group(32 cases)and non-HH group(202 cases).The total incidence of HH was 13.7%(32/234).Septic shock was the main disease of the occurrence of HH in children with shock,accounting for 65.63%(21/32).According to the prognosis,children with shock combined HH were divided into survival group(17 cases)and death group(15 cases).The case fatality rate of children with shock combined HH was 46.9%.The ALT and AST values in children with shock combined HH were more than 20 times of the upper limits of normal ranges.The peak values of enzymatic indexes in the survival group appeared at the onset time d 1 to d 2,they were close to the normal ranges at the onset time d7,while the enzymatic indexes in the death group were higher than the normal ranges during the whole disease stage.Compared with the survival group,the peak values of ALT,AST,ALP,TBIL,CRP,BUN,and INR in the death group were obviously higher(P<0.05,both).The lowest value of ALB in the death group was lower than that in the survival group(P<0.05).The results of multivariate Logistic regression analysis showed that pediatric critical illness score(PCIS),lactate,multiple organ dysfunction syndrome(MODS),decompensated shock and cardiogenic shock were the risk factors for the occurrence of HH in children with shock(P<0.05 or 0.01),and left ventricular ejection fraction(LVEF)and mean arterial pressure(MAP)were the risk factors of the death of HH in children with shock(P<0.01).Conclusions Children with shock who have the risk factors as decreased PCIS,increased lactate level,MODS,decompensated shock and cardiogenic shock are apt to suffer from HH.Poor prognosis in the chil-dren with shock combined HH is associated with worse condition and lower LVEF.There is a high fatality rate of shock combined HH in children,so that liver function indicators should be closely monitored to early detect HH,and reasonable treatment should be given.

Pancreatic cancer is among the most malignant cancers,and thus early intervention is the key to better survival outcomes.However,no methods have been derived that can reliably identify early precursors of development into malignancy.Therefore,it is urgent to discover early molecular changes during pancreatic tumorigenesis.As aberrant glycosylation is closely associated with cancer progression,numerous efforts have been made to mine glycosylation changes as biomarkers for diagnosis;however,detailed glycoproteomic information,especially site-specific N-glycosylation changes in pancreatic cancer with and without drug treatment,needs to be further explored.Herein,we used comprehensive solid-phase chemoenzymatic glycoproteomics to analyze glycans,glycosites,and intact glycopeptides in pancreatic cancer cells and patient sera.The profiling of N-glycans in cancer cells revealed an increase in the secreted glycoproteins from the primary tumor of MIA PaCa-2 cells,whereas human sera,which contain many secreted glycoproteins,had significant changes of glycans at their specific glycosites.These results indicated the potential role for tumor-specific glycosylation as disease biomarkers.We also found that AMG-510,a small molecule inhibitor against Kirsten rat sarcoma viral oncogene homolog(KRAS)G12C mutation,profoundly reduced the glycosylation level in MIA PaCa-2 cells,suggesting that KRAS plays a role in the cellular glycosylation process,and thus glycosylation inhibition contributes to the anti-tumor effect of AMG-510.

Objective:To research the association between exposure to solid fuels for heating and its duration and the risk of respiratory diseases morbidity.Methods:Data from the China Kadoorie Biobank project sited in Pengzhou City, Sichuan Province. Cox proportional hazard regression model was used to analyze the association between exposure to solid fuels for heating and its duration and the risk of total respiratory diseases and the association between exposure to solid fuels for heating and the risk of chronic obstructive pulmonary disease (COPD) and pneumonia among respiratory diseases.Results:A total of 46 082 participants aged 30-79 years were enrolled, with 11 634 (25.25%) heating during the winter, of whom 8 885 (19.28%) used clean fuels and 2 749 (5.97%) used solid fuels, of whom 34 448 (74.75%) did not heat. After controlling for multiple confounding factors, Cox proportional hazard regression model was used, which revealed that compared with clean fuels, unheating could reduce the risk of total respiratory disease ( HR=0.81,95% CI:0.77-0.86), COPD ( HR=0.86,95% CI:0.78-0.95) and pneumonia ( HR=0.80,95% CI:0.74-0.86), respectively. Exposure to solid fuels increased the risk of total respiratory disease ( HR=1.10, 95% CI:1.01-1.20) and were not associated with COPD and pneumonia. Compared with no solid fuel exposure, the risk of total respiratory disease (1-19 years: HR=1.23, 95% CI:1.10-1.37; 20-39 years: HR=1.25, 95% CI:1.16-1.35; ≥40 years: HR=1.26, 95% CI:1.15-1.39) and COPD (1-19 years: HR=1.21, 95% CI:1.03-1.42; 20-39 years: HR=1.30, 95% CI:1.16-1.46; ≥40 years: HR=1.35, 95% CI:1.18-1.54) increased with the length of exposure of solid fuels (trend test P<0.001). Solid fuels exposure for 1-19 years and 20-39 years increased the risk of COPD by 23% ( HR=1.23,95% CI:1.02-1.49) and 16% ( HR=1.16, 95% CI:1.00-1.35). Conclusion:Heating solid fuels exposure increases the risk of total respiratory disease, COPD, and pneumonia.

Objective:To investigate the morbidity of cerebrovascular disease among residents ≥30 years in Pengzhou, Sichuan Province, and analyze the effect of physical activity level on the risk of morbidity of cerebrovascular disease.Methods:From 2004 to 2008, people from Pengzhou, Sichuan Province were randomly selected. All the local people aged 30-79 were asked to receive a questionnaire survey, physical examination, and long-term follow-up to determine the morbidity of cerebrovascular disease. The physical activity level and the morbidity of cerebrovascular disease were described, and Cox proportional hazard regression models were used to evaluate the association of domain-specific physical activity with the risk of morbidity of cerebrovascular disease.Results:In 55 126 participants, there were 5 290 new cases of cerebrovascular disease, with a cumulative incidence of 9.60%. After the adjustment for multiple confounding factors, multivariate Cox proportional hazard regression analysis showed that increased levels of occupational, transportation, and total physical activity reduced the risk of cerebrovascular disease and its subtypes (cerebral hemorrhage, cerebral infarction). The highest group of occupational physical activity level had the lowest risk of cerebrovascular disease, with a hazard ratio ( HR) value of 0.81 (95% CI: 0.75-0.88), the highest group of transportation physical activity level had the lowest risk of cerebrovascular disease, with an HR value of 0.84 (95% CI: 0.78-0.91), the highest group of total physical activity level had the lowest risk of cerebrovascular disease, with an HR value of 0.87 (95% CI: 0.80-0.94), compared with the lowest group of corresponding physical activity. No association was found between the household/leisure-time physical activity level and the risk of cerebrovascular disease and its subtypes (cerebral hemorrhage, cerebral infarction). Conclusions:In project areas of Pengzhou, Sichuan Province, increased physical activity has been associated with reduced morbidity of cerebrovascular disease and its subtypes (cerebral hemorrhage, cerebral infarction). Increased levels of physical activity in adults are encouraged for health benefits.

Objective:To understand the infection status, epidemiological characteristics and drug resistance of Diarrheagenic Escherichia coli (DEC) in Shanghai and provide evidence for the disease surveillance. Methods:The epidemiological data of diarrhea cases in Shanghai from 2016 to 2022 were collected from Shanghai Diarrhea Comprehensive Surveillance System, and stool samples were collected from the cases for DEC detection. The drug resistance data was obtained from Chinese Pathogen Identification Network. Statistical analysis was conducted by using χ2 and fisher test. Results:In 24 883 diarrhea cases detected during 2016-2022, the DEC positive rate was 9.13% (2 271/24 883), the single DEC positive rate was 8.83% (2 197/24 883) and the mixed DEC positive rate was 0.30% (74/24 883). The main type of DEC was Enterotoxigenic Escherichia coli (ETEC) [4.33% (1 077/24 883)]. The DEC positive rate was highest in people aged ≤5 years 18.48% (22/119). The annual peak of DEC positive rate was observed during July - September [5.91% (1 470/24 883)]. The DEC positive rate were 9.47% (554/5 847) and 9.02% (1 717/19 036) in urban area and in suburbs, respectively, Enteroaggregative Escherichia coli (EAEC) [3.98% (233/5 847)] and ETEC [4.56% (868/19 036)] were mainly detected. From 2016 to 2019, the DEC positive rate was 9.42% (1 821/19 330), while it was 8.10% (450/5 553) from 2020 to 2022, the main DEC types were ETEC (4.87%, 941/19 330) and EAEC (4.70%, 261/5 553). The multi-drug resistance rate was 40.21% (618/1 537). The top three antibiotics with high drug resistance rates were ampicillin [64.74% (995/1 537)], nalidixic acid [58.49% (899/1 537)] and tetracycline [45.09% (693/1 537)]. Conclusions:Compared with 2016- 2019, a decrease in DEC detection rate was observed during 2020-2022, and the main type of DEC detected shifted from ETEC to EAEC. The prevalence of multi-drug resistance was severe. Therefore, it is necessary to further strengthen the surveillance for DEC drug resistance and standardize the use of clinical antibiotics.