To systematically analyzed the reporting status of core elements in publicly available clinical practice guideline(hereafter referred to as "guideline") protocols published domestically and internationally over the past decade, identified existing problems, and provided evidence to inform the standardized writing and publication of future guideline protocols.

Objective: To analyze the impact of premature death due to four major chronic diseases on life expectancy in Yangpu District, Shanghai Municipality from 2010 to 2021, so as to provide the evidence for formulating chronic disease prevention and control strategies. Methods : Mortality data of registered residents in Yangpu District from 2010 to 2021 were collected through the Death Information Registration and Management System of the Shanghai Municipal Disease Control and Prevention Information Management Platform. The premature death probability of malignant tumors, diabetes, cardiovascular and cerebrovascular diseases, and chronic respiratory diseases, and life expectancy of residents were calculated using the abridged life table method. Trends in premature death probability for four types of chronic diseases were analyzed using the average annual percent change (AAPC). The impact of premature death probability due to four chronic diseases on life expectancy was assessed by Arriaga's decomposition method. Results : The premature death probability due to four major chronic diseases in Yangpu District decreased from 9.88% in 2010 to 9.22% in 2021, showing an overall declining trend (AAPC=-0.540%, P<0.05). Among females, the premature death probability declined from 6.71% to 4.90% (AAPC=-2.715%, P<0.05), whereas no statistically significant trend was observed in males (P>0.05). Life expectancy increased from 82.52 years in 2010 to 84.50 years in 2021, with an overall upward trend (AAPC=0.244%, P<0.05). Life expectancy rose by 1.71 years for males and 2.34 years for females (AAPC=0.197% and 0.303%,both P<0.05). Declines in premature death probability from malignant tumors (AAPC=-0.967%, P< 0.05) and chronic respiratory diseases (AAPC=-3.071%, P<0.05) contributed to gains in life expectancy of 0.30 years and 0.03 years, with contribution rates of 12.18% and 1.29%, respectively. Changes in premature death probability due to diabetes as well as cardiovascular and cerebrovascular diseases were not statistically significant (both P>0.05), resulting in reductions in life expectancy of 0.05 years and 0.10 years, with contribution rates of -2.40% and -5.05%, respectively. Notably, an increase in premature death probability due to cardiovascular and cerebrovascular diseases among males (AAPC=1.673%) contributed to a decrease of 0.22 years in male life expectancy, whereas a decrease among females (AAPC=-3.824%) contributed to an increase of 0.03 years in female life expectancy, with contribution rates of -13.03% and 1.14%, respectively. Conclusions From 2010 to 2021, Yangpu District experienced an overall decline in premature death probability due to four major chronic diseases and an increase in life expectancy. Greater attention should be paid to the negative impacts of premature death probability from diabetes as well as cardiovascular and cerebrovascular diseases among males on life expectancy.

Objective To explore the effects of individualized nutrition intervention combined with one-day outpatient mode on blood glucose management and pregnancy outcomes in pregnant women with gestational diabetes mellitus(GDM).Methods A total of 395 pregnant women diagnosed with GDM who underwent prenatal examination in Maternity and Child Health Care of Guangxi Zhuang Autonomous Region from January 2023 to January 2024 were selected as study objects.According to the nutritional intervention measures,the pregnant women were divided into control group(102 cases),nutrition intervention group(141 cases)and combined intervention group(152 cases).The control group was given routine diet education,and nutrition intervention group received individualized nutrition intervention on the basis of control group,and combined intervention group received one-day outpatient intervention on the basis of nutrition intervention group.All pregnant women in three groups were intervened until delivery.The general data,blood glucose indexes and pregnancy outcome of three groups were compared.Results After the intervention,the levels of preprandial blood glucose and 2h postprandial blood glucose on the day of delivery,glycated hemoglobin in late pregnancy and postpartum fasting blood glucose in combined intervention group were significantly lower than those in nutrition intervention group and control group(P＜0.05).The levels of preprandial blood glucose on the day of delivery and postpartum fasting blood glucose in nutrition intervention group were significantly lower than those in control group(P＜0.05).The incidence of preterm birth,low birth weight infants,neonatal hyperbilirubinemia and neonatal hypoglycemia in nutrition intervention group and combined intervention group were significantly lower than those in control group(P＜0.05).Conclusion Individualized nutrition intervention combined with one-day outpatient mode can help to manage blood glucose in pregnant women with GDM and improve pregnancy outcome.

Objective:To explore the distribution of pathogenic bacteria causing postoperative infections in chil-dren undergoing surgery for Hirschsprung's disease(HD),and the clinical significance of serum triggering receptor ex-pressed on myeloid cells-1(sTREM-1)and high mobility group box 1(HMGB1).Methods:98 children with HD admit-ted from January 2021 to January 2025 were divided into infection group(n=42)and non-infection group(n=56)accord-ing to whether the children were complicated with infection after operation.Automatic bacterial identification system was used to detect the distribution of pathogenic bacteria in secretion samples from infected sites of HD children after operation.ELISA was used to measure serum sTREM-1 and HMGB1.ROC was used to explore the diagnostic value of serum sTREM-1 and HMGB1 for severe infection in children with HD.Results:A total of 49 strains of pathogenic bac-teria were isolated from 42 children with HD in infection group,of which 63.27%(31/49)were gram-negative bacteria,accounting for the highest proportion,followed by Gram positive bacteria at 30.61%(15/49)and fungi at 6.12%(3/49).Among them,Pseudomonas aeruginosa had the highest proportion of 24.49%(12/49)among Gram negative bacteria.there were 15 cases of mild infection(15 strains of pathogens),16 cases of moderate infection(17 strains of pathogens)and 11 cases of severe infection(17 strains of pathogens).With the aggravation of the infection degree of HD children,the distribution pattern of pathogenic bacteria changed,and gradually changed to mainly gram-negative bacteria infec-tion.Compared with non infected group,the levels of serum sTREM-1 and HMGB1 in infected group were higher(P<0.05).The levels of serum sTREM-1 and HMGB1 in children with severe HD infection are higher than those in children with mild and moderate HD infection(P<0.05).The AUC of the combined diagnosis of serum sTREM-1 and HMGB1 for severe infection in HD children was the highest,at 0.985,which was superior to the separate diagnosis of serum sTREM-1 and HMGB1(Zcombination-sTREM-1=1.988,P=0.047;Zcombination-HMGB1=2.126,P=0.034).Conclusion:Serum sTREM-1 and HMGB1 are elevated in HD children with postoperative infection,and the pathogenic bacteria are mainly gram-negative bacteria.In addition,the combined detection of the two provides a reference for the clinical diagnosis of severe infection in HD children.

Objective:To explore the application effects of the blended teaching methodology of "teaching-selection-investigation-analysis-presentation-discussion" in Medical Immunology. Methods:Eight-year program clinical medical students who were enrolled at Peking Union Medical College in 2016, 2017, and 2018 were selected as the research subjects. An anonymous questionnaire survey was used to analyze students' multidimensional evaluations of the new teaching methodology. The percentage of frontier hotspot topics in the "Immunology Forum" was used to analyze the students' mastery of cutting-edge knowledge in immunology. The number of "Immunology Forum" related "College Students Innovative Training Plan Program" from 2020 to 2023 was used to analyze the effectiveness of this new teaching method in cultivating students' scientific research and innovation abilities. Statistical analysis was conducted using SPSS 25.0 software, and the normality of all continuous variables was assessed using the Shapiro-Wilk test.Results:The questionnaire survey showed 100.00% satisfaction with the course and 95.90% recognition of the new teaching method. More than 90% of students agreed that the new teaching method improved their learning ability, research ability, innovation ability, internal drive, and academic communication ability. The average proportion of hotspot topics in the "Immunology Forum" was 90.37%±7.12%, which was significantly higher than the proportion of non-hotspot topics (5.67%±3.12%). The average number of topics related to "Immunology Forum" in the "College Students Innovative Training Plan Program" was 17.67±1.15 per session, which was significantly higher than the number of topics not related to "Immunology Forum" (8.00±1.73).Conclusions:The blended teaching methodology of "teaching-selection-investigation-analysis-presentation-discussion" can help students timely grasp the cutting-edge knowledge of immunology, cultivate their learning ability, internal drive, academic communication ability, innovation ability, and research ability, and lay a foundation for students to further explore their scientific research and innovation activities.

AIM To study the chemical constituents from the stems and leaves of Dendrobium formosum Roxb.ex Lindl.and their biological activities.METHODS The 95％ethanol extract from the stems and leaves of D.formosum was isolated and purified by silica gel,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.Their inhibitory activities onα-glucosidase were determined by PNPG method,and their in vitro anti-inflammatory activities were evaluated by RAW264.7 model.RESULTS Fifteen compounds were isolated and identified as coniferyl p-coumarate(1),(-)-pinoresinol(2),2,5,7-trihydroxy-4-methoxy-9,10-dihydrophenanthrene(3),naringenin(4),spiropreussomerin A(5),7-hydroxy-14-de-O-methyl-lasiodiplodin(6),(4S,5S,6Z,8E)-5-hydroxydeca-6,8-dien-4-olide(7),(6S,9R)-blumenol C(8),p-hydroxybenzoic acid(9),m-hydroxybenzoic acid(10),p-hydroxy benzenepropanoic acid(11),5,7-dihydroxy-isobenzofuran(12),2-(4-hydroxyphenyl)-ethanol(13),β-sitostenone(14),β-sitosterol(15).The IC50 values of compounds 1 and 4 on α-glucosidase inhibition were(65.60±3.31)and(98.95±2.53)μmol/L,respectively.Compound 3 presented inhibitory activity on NO production in RAW 264.7 cells,with IC50 value of(3.97±0.12)μmol/L.CONCLUSION Compounds 5-6,8 and 12 are isolated from Orchidacae family for the first time,and 2-15 are first isolated from this plant.Compounds 1 and 4 have α-glucosidase inhibitory activities,and 3 has anti-inflammatory activity.

Aim To investigate the neuroprotective effect of Takeda G protein-coupled receptor-5(TGR5)activated by INT-777 on hypoxic-ischemic encephalop-athy(HIE)in neonatal rats.Methods Seven-day-old SD rats were randomly divided into the sham opera-tion group(Sham,S),model group(HIE,G),INT-777 low-dose(L),medium-dose(M),and high-dose(H)groups.The modified Rice-Vanucci method was used to construct the HIE model and Intranasal admin-istration 1 h after modeling.Short-term neurobehavioral tests were performed 48 h after modeling to evaluate the neurological function of neonatal rats,TTC staining was used to determine the volume of cerebral infarction,dry and wet specific gravity was used to determine the brain water content,ferrous ion kit was used to deter-mine the brain ferrous ion content,HE staining was used to observe the pathological damage of brain tis-sue,Nissl staining was used to observe the loss of Nissl substance,Transmission electron microscopy(TEM)was used to observe the mitochondrial morphological changes of cortical neurons,and Western blot was em-ployed to detect the expression of ferroptosis-related proteins TFR1 and GPX4.Results Compared with group S,group G had increased short-term neurobehav-ioral test consumption time,higher scores,increased cerebral infarct volume,brain water content,and brain ferrous iron content,significant brain tissue damage on the affected side,severe loss of Nissl substance,smaller neuronal mitochondria,decreased mitochondrial cris-tae,and increased expression of TFR1 and reduced ex-pression of GPX4.Compared with group G,the INT-777 administration group had a shorter consumption time for short-term neurobehavioral tests,lower scores,the cerebral infarction volume,brain water content,and brain ferrous ion content decreased,the brain tissue damage on the affected side was reduced,and there was insignificant loss of Nissl substance,larger neuronal mi-tochondrial volume,increased mitochondrial cristae,re-duced expression of TFR1,and increased expression of GPX4.Conclusions INT-777 agonist TGR5 has a protective effect against hypoxic-ischemic encephalopa-thy in neonatal rats,and its mechanism of action may be related to the inhibition of neuronal ferroptosis.

Objective To explore the effects of individualized nutrition intervention combined with one-day outpatient mode on blood glucose management and pregnancy outcomes in pregnant women with gestational diabetes mellitus(GDM).Methods A total of 395 pregnant women diagnosed with GDM who underwent prenatal examination in Maternity and Child Health Care of Guangxi Zhuang Autonomous Region from January 2023 to January 2024 were selected as study objects.According to the nutritional intervention measures,the pregnant women were divided into control group(102 cases),nutrition intervention group(141 cases)and combined intervention group(152 cases).The control group was given routine diet education,and nutrition intervention group received individualized nutrition intervention on the basis of control group,and combined intervention group received one-day outpatient intervention on the basis of nutrition intervention group.All pregnant women in three groups were intervened until delivery.The general data,blood glucose indexes and pregnancy outcome of three groups were compared.Results After the intervention,the levels of preprandial blood glucose and 2h postprandial blood glucose on the day of delivery,glycated hemoglobin in late pregnancy and postpartum fasting blood glucose in combined intervention group were significantly lower than those in nutrition intervention group and control group(P＜0.05).The levels of preprandial blood glucose on the day of delivery and postpartum fasting blood glucose in nutrition intervention group were significantly lower than those in control group(P＜0.05).The incidence of preterm birth,low birth weight infants,neonatal hyperbilirubinemia and neonatal hypoglycemia in nutrition intervention group and combined intervention group were significantly lower than those in control group(P＜0.05).Conclusion Individualized nutrition intervention combined with one-day outpatient mode can help to manage blood glucose in pregnant women with GDM and improve pregnancy outcome.

Aim To investigate the neuroprotective effect of Takeda G protein-coupled receptor-5(TGR5)activated by INT-777 on hypoxic-ischemic encephalop-athy(HIE)in neonatal rats.Methods Seven-day-old SD rats were randomly divided into the sham opera-tion group(Sham,S),model group(HIE,G),INT-777 low-dose(L),medium-dose(M),and high-dose(H)groups.The modified Rice-Vanucci method was used to construct the HIE model and Intranasal admin-istration 1 h after modeling.Short-term neurobehavioral tests were performed 48 h after modeling to evaluate the neurological function of neonatal rats,TTC staining was used to determine the volume of cerebral infarction,dry and wet specific gravity was used to determine the brain water content,ferrous ion kit was used to deter-mine the brain ferrous ion content,HE staining was used to observe the pathological damage of brain tis-sue,Nissl staining was used to observe the loss of Nissl substance,Transmission electron microscopy(TEM)was used to observe the mitochondrial morphological changes of cortical neurons,and Western blot was em-ployed to detect the expression of ferroptosis-related proteins TFR1 and GPX4.Results Compared with group S,group G had increased short-term neurobehav-ioral test consumption time,higher scores,increased cerebral infarct volume,brain water content,and brain ferrous iron content,significant brain tissue damage on the affected side,severe loss of Nissl substance,smaller neuronal mitochondria,decreased mitochondrial cris-tae,and increased expression of TFR1 and reduced ex-pression of GPX4.Compared with group G,the INT-777 administration group had a shorter consumption time for short-term neurobehavioral tests,lower scores,the cerebral infarction volume,brain water content,and brain ferrous ion content decreased,the brain tissue damage on the affected side was reduced,and there was insignificant loss of Nissl substance,larger neuronal mi-tochondrial volume,increased mitochondrial cristae,re-duced expression of TFR1,and increased expression of GPX4.Conclusions INT-777 agonist TGR5 has a protective effect against hypoxic-ischemic encephalopa-thy in neonatal rats,and its mechanism of action may be related to the inhibition of neuronal ferroptosis.

OBJECTIVE To screen and obtain nanobodies with neutralizing activity against granulo-cyte-macrophage colony stimulating factor(GM-CSF)to contribute to investigations into the mecha-nism of inflammation interventions and the development new drugs.METHODS Recombinant human GM-CSF was subcutaneously injected to immunize camels.Peripheral blood was collected after five immunizations,and mononuclear cells were isolated.Total mRNA was extracted,and the variable domains of the heavy chain of heavy-chain antibody(VHH,also called nanobody)genes were obtained by PCR amplification after reverse transcription.The genes were cloned into the pADSCFV-S phage display vector and electrotransformed into TG1 competent cells to construct a nanobody immune library that was screened with recombinant human GM-CSF immobilized on a solid phase.The VHH genes specifi-cally binding to human GM-CSF were cloned into the pABG eukaryotic expression vector before VHH-Fc samples were prepared by using the human embryonic kidney 293 fibroblast expression system.The binding activity of candidate VHH-Fc molecules to GM-CSF was investigated through ELISA response curves,and binding colorimetric values with different antigens were detected to determine their specificity.The binding affinity between VHH-Fc candidates and GM-CSF was measured using biolayer interferom-etry(BLI).The inhibition rate curve of VHH-Fc candidates on GM-CSF was detected through cell prolif-eration assays to determine its neutralization activity.The Uncle system was used to investigate its thermal stability.100 μg of VHH-Fc was injected into mice via the tail vein,and the serum concentration of VHH-Fc was quantitatively detected by ELISA to examine its pharmacokinetic curve in mice.RESULTS The camel serum titer of anti-human GM-CSF antibodies was higher than 1:800 000 after the fifth immuni-zation,and the capacity of the constructed nanobody library was about 5.55×107.Following the screening process,five candidate VHH-Fc molecules specifically binding to human GM-CSF were obtained.Among these,22N10 effectively neutralized the cell proliferation-promoting activity of GM-CSF(the IC50 value was 17.23 nmol·L-1).Subsequent studies revealed that 22N10 interacted with human GM-CSF with an affinity of 1.97×10-8 mol·L-1,blocked the binding of GM-CSF to its receptor CSF2Rα,exhibited good thermal stability(Tm1=59.2℃),and showed favorable metabolic characteristics in mice.CONCLU-SION A new candidate nanobody molecule 22N10 targeting human GM-CSF is obtained which is expected to facilitate the drug development and mechanism investigations.