OBJECTIVE To analyze and identify the metabolites of pirtobrutinib （PTN） in rats， and clarify the possible metabolic pathways of PTN in rats. METHODS Six rats were intragastrically administered with 10 mg/kg PTN suspension. Blood samples were collected from the rats 30 minutes before administration and at 0.25， 0.5， 1， 2， 4， 6， 8， 12， 24 hours after administration. Urine and feces samples were collected 12 hours before administration and 24 hours after administration. UHPLC- Orbitrap Exploris 240 system combined with Compound Discoverer 3.0 and Xcalibur 2.0 software were adopted for structural identification and metabolic pathway analysis of PTN metabolites in rat plasma， urine， and feces. RESULTS A total of 29 PTN metabolites were identified， including 17， 19 and 22 metabolites in plasma， urine and feces， respectively. The metabolic pathways of PTN mainly included oxidation， sulfation， glucuronidation， etc.， and its metabolites were mostly combination products of two or more different metabolic forms. In detail， a total of 26 metabolites were associated with phase Ⅰ metabolic reactions （14 oxidation metabolites， 9 reduction/dehydrogenation metabolites， 8 demethylation metabolites， and 5 hydrolysis metabolites）. Meanwhile， a total of 20 products were involved in phase Ⅱ metabolites （14 sulfation metabolites and 8 glucuronic acid binding metabolites）. CONCLUSIONS PTN exhibits a diverse range of metabolites in rat fecal samples， with the primary metabolic pathways being oxidation， sulfation， glucuronidation， and others.

Objective:To investigate the impact of daily step count on glycemic outcomes in community residents with impaired glucose tolerance (IGT).Methods:This was a prospective cohort study, in October 2018, 204 residents who met the criteria of IGT were recruited in the Shijingshan District in Beijing. The subjects were tested for fasting blood glucose, oral glucose tolerance test 2-hour blood glucose (2hBG), glycated hemoglobin A 1c (HbA 1c), lipid profile, liver and kidney function, as well as measurements of height, weight and waist circumference. A dedicated mobile application was used to deliver prediabetes health knowledge monthly. Online guidance was provided to answer questions and daily step count was collected using the application. Three years later, a follow-up was conducted to assess the participants′ glycemic outcomes and other indexes, and a total of 142 participants completed the follow-up review. According to daily step count, the subjects were categorized into high step count group (42 cases,>7 000 steps daily), moderate step count group (54 cases, 5 000-7 000 steps daily), and low step count group (46 cases,<5 000 steps daily). Subjects were categorized into diabetes group (30 cases), prediabetes group (77 cases) and normal glucose tolerance group (35 cases) with glycemic outcomes. Independent sample t test was used to compare the differences in blood glucose, blood lipids, and step counts between the two groups. Kruskal-Wallis H test or one-way ANOVA was used to compare the differences in blood glucose, blood lipids, and step counts between multiple groups. The χ2 test was used to compare the differences in glycemic outcomes between multiple groups. Multivariate logistic regression analysis was used to assess the impact of daily step counts and body mass index on glycemic outcomes. Linear regression analysis was used to evaluate the relationship between daily step counts and 2 h BG. Results:A total of 142 participants completed the 3-year follow-up, including 43 males and 99 females, with a mean age of (60.15±5.67) years. At baseline, males had significantly higher body mass index, waist circumference, and fasting blood glucose when compared to those in females [(26.97±2.43) vs (24.89±2.93) kg/m 2, (92.68±7.75) vs (83.83±8.60) cm, (5.83±0.61) vs (5.62±0.52) mmol/L], the total cholesterol and HDL-C were also significantly lower in males than those in females [(5.10±1.16) vs (5.55±0.95) mmol/L, (1.35±0.34) vs (1.56±0.35) mmol/L] (all P<0.05). After 3-year follow-up, 21.1% (30/142) of IGT participants progressed to diabetes, with an annual conversion rate of approximately 7%. The normal glucose tolerance group showed significantly higher daily step counts when compared with the prediabetes and diabetes groups [(7 886±2 867) vs (5 981±2 655) vs (4 117±2 674) steps] ( H=31.778, P<0.001). Individuals with higher daily step counts exhibited lower body mass index, 2 h BG, and HbA 1c level when compared with those in the ones with moderate and low step counts [(24.26±3.09) vs (25.44±3.38) vs (26.26±3.59) kg/m 2, (7.50±1.71) vs (9.15±3.30) vs (11.19±3.84) mmol/L, 5.97%±0.46% vs 6.14%±0.99% vs 6.40%±0.96%] (all P<0.05). Higher step count was positively correlated with the reversal of prediabetes to normal blood glucose levels (moderate step count, OR=0.297, 95% CI: 0.109-0.804; low step count, OR=0.055, 95% CI: 0.010-0.287), lower daily step count correlated positively with prediabetes progressing to diabetes ( OR=4.857, 95% CI: 1.140-20.689) (all P<0.05). For every additional 1 000 steps per day, the 2 h BG decreased by 0.5 mmol/L. Conclusion:As daily step count increases, the glucose metabolism improves in IGT community residents. Higher daily step count is associated with reversal of IGT to normal glucose tolerance, while lower daily step count may be associated with the progression of IGT to diabetes.

Objective:To assess the effects of low temperature environment on skeletal and serum metabolites, and explore underlying mechanism.Methods:C57BL/6J mice were divided into normal temperature control group(control group) and intermittent low temperature exposure group(low temperature group). After 12 weeks of intervention, Micro-computed tomography(CT) was used to detect the bone microstructure. The mouse femur was stained by hematoxylin-eosin(HE) staining, tartrate-resistant acid phosphase(TRAP) staining, and type Ⅰ collagen staining. The serum metabolites were detected by liquid chromatography-mass spectrometry(LC-MS), comprehensive principal component analysis and random forest were used to identify potential biomarkers influencing bone metabolism in serum samples, and the metabolic pathways affecting bone metabolism were enriched through metabolic database analysis.Results:Micro-CT results showed that compared to the control group, the bone mineral density, bone volume/tissue volume, trabecular thickness, and trabecular number decreased(all P<0.05) in low temperature group of mice, while trabecular separation and bone surface/bone volume ratio increased(both P<0.05). The staining results of HE indicated that, compared with the control group, low temperature group exhibited disordered bone trabecular space structure, widened gaps, reduced unit area, and the presence of numerous adipose vacuoles. TRAP staining suggested that compared to the control group, low temperature group had an increased number of osteoclasts. The results of type Ⅰ collagen staining showed that the number of type Ⅰ collagen in the low temperature group was lower than that in the control group, and the structure was disordered. On the other hand, metabolomics identified 40 metabolites in serum, including deoxycholic acid, methionine, bilirubin, and salicylic acid, which are involved in lipid metabolism and amino acid metabolism. Conclusion:Low temperature leads to decreased bone mass, which may be related to the regulation of lipid metabolism and amino acid metabolism.

Objective:In order to explore the impact of corona virus disease 2019(COVID-19)on the hospitalization of children with bronchiolitis and to improve clinicians′ understanding of the characteristics of bronchiolitis during the COVID-19 epidemic.Methods:This was a multicenter clinical study, and the data have been collected from 23 children′s medical centers in China.All the clinical data were retrospectively collected from children with bronchiolitis who were hospitalized at each study center from January 1, 2019 to December 31, 2021.The results included gender, age at hospitalization, length of stay, respiratory syncytial virus(RSV) test results, severity rating, ICU treatment, and the total number of children hospitalized with respiratory tract infection during the same period.The clinical data of children with bronchiolitis in 2019 before COVID-19 epidemic and in 2020、2021 during COVID-19 epidemic were statistically analyzed and compared.Results:According to a summary of data provided by 23 children′s medical centers, there were 4 909 cases of bronchiolitis in 2019, 2 654 cases in 2020, and 3 500 cases in 2021.Compared with 2019, the number of bronchiolitis cases decreased by 45.94% in 2020 and 28.70% in 2021.In 2019, 2020 and 2021, there were no significant differences in gender ratio, age, and duration of hospitalization.Compared with 2019, the ratio of bronchiolitis to the total number of hospitalizations for respiratory tract infection decreased significantly in 2020 and 2021( χ2=12.762, P<0.05; χ2=84.845, P<0.05).The proportion of moderate to severe bronchiolitis cases in both 2020 and 2021 was lower than that in 2019, and the difference was statistically significant ( χ2=4.054, P<0.05; χ2=8.109, P<0.05).There was no statistically significant difference in the proportion of bronchiolitis cases requiring ICU treatment between 2019, 2020, and 2021 ( χ2=1.914, P>0.05).In 2019, a total of 52.60%(2 582/4 909) of children with bronchiolitis underwent RSV pathogen testing, and among them, there were 708 cases with RSV positive, accounting for 28.00%.In 2020, 54.14%(1 437/2 654) of children with bronchiolitis underwent RSV pathogen testing, and there were 403 cases with RSV positive, accounting for 28.04%.In 2021, 66.80%(2 238/3 500) of children with bronchiolitis underwent RSV pathogen testing, and there were 935 cases with RSV positive, accounting for 41.78%.Compared with 2019 and 2020, the RSV positive rate in 2021 showed a significant increase( χ2=99.673, P<0.05; χ2=71.292, P<0.05). Conclusion:During the COVID-19 epidemic, the implementation of epidemic prevention and control measures reduced the hospitalization rate and severity of bronchiolitis, but did not reduce the positive rate of RSV detection.

Objective:To evaluate the effect of febutostat on vascular endothelial function, intima-media thickness(C-IMT) and elasticity of the carotid artery in patients with asymptomatic hyperuricemia.Methods:This study was a randomized controlled clinical trial that enrolled asymptomatic hyperuricemia patients from the outpatient and inpatient departments of Huai′an First People′s Hospital from October 2018 to October 2020. The participants were randomly divided into two groups: the Febuxostat group and the control group. Serum triglyceride(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), fasting blood glucose(FBG), fasting insulin(FINS), nitric oxide(NO), endothelin-1(ET-1), malondialdehyde(MDA), and superoxide dismutase(SOD) were measured at baseline and 1, 3, and 6 months after treatment, and brachial artery flow-mediated dilation(FMD) was quantified by color Doppler ultrasound. The following parameters of the common carotid artery were detected at baseline and 12 months after treatment: C-IMT, arterial compliance(AC), one-point pulse wave velocity(PWV), stiffness index(β), and pressure-strain elasticity modulus(Ep). The differences before and after treatment and between the two groups were compared. Pearson correlation was used to analyze the correlation between ΔUA and ΔNO, ΔET-1, ΔC-IMT, ΔAC, Δβ, ΔEp, and ΔPWVβ after treatment with febuxostat. Results:Compared with baseline, TG, HOMA-IR, ET-1 and MDA were significantly lower, while FMD, NO and SOD were significantly higher after 3-months treatment with febuxostat. After 12-months treatment, there was no significant difference in C-IMT or Ep, but there was an increase in AC and a decrease in PWVβ or β compared with baseline. There was a negative correlation between ΔFMD and ΔUA( r=-0.403, P=0.004), but there were no correlations between ΔNO and ΔUA( r=-0.187, P=0.194), ΔET-1 and ΔUA( r=0.038, P=0.791) after 6-months treatment. And ΔUA was an independent factor for ΔFMD( F=2.94, P=0.003, adjusted R2=0.139). After 12-months treatment, there was a negative correlation between ΔAC and ΔUA, and a positive correlation between ΔPWVβ and ΔUA, but there were no correlations between the following indicators: ΔC-IMT and ΔUA( r=0.169, P=0.240), Δβ and ΔUA( r=-0.214, P=0.136), ΔEp and ΔUA( r=-0.077, P=0.597). In the control group, there were no differences among the above indicators between each follow-up time and baseline. Conclusion:Febuxostat improves vascular endothelial function and elasticity in patients with asymptomatic hyperuricemia, which may be related to the decreased oxidative stress response.

Objective: To explore the high expression level of serum heat shock protein 90α(HSP90α) and gene HSP90 AA1 in cancer tissue, and to discover the prognosis of lung cancer. Methods: A total of 109 cases of lung cancer were collected as the experimental group; 38 lung inflammation groups as the reference group; and 30 healthy controls. The serum HSP90α levels between the three groups were compared; the correlation between HSP90α and clinical parameters were analyzed. The TCGA data were used to analyze the correlation between the expression level of HSP90 AA1 and various pathological features, as well as its influence on the prognosis of lung cancers. Results: The serum HSP90α concentrations in the experimental group were higher than those of the reference group and the control group(P<0.05). The ROC curve area of HSP90α in the diagnosis of lung cancer was 0.898(P<0.05); the expression of HSP90α in the lung squamous cell carcinoma group(LUSC) and the small cell lung cancer group(SCLC) were higher than those in lung adenocarcinoma group(LUAD)(P<0.05); the level of HSP90α decreased when condition alleviated, while increased significantly when disease progressed(P<0.05); Further TCGA database showed that the expression of HSP90 AA1 in cancer tissues was higher than that of adjacent cancer tissues(P<0.05),meanwhile, remarkably increased in LUSC compared with in LUAD(P<0.05). The expression of HSP90 AA1 has no significant correlation with the age, gender, clinical stage, and tumor residue of lung cancer patients. Survival analysis and Cox regression analysis showed that high expression of HSP90 AA1 reduced the overall survival(OS) of lung cancer patients; HSP90 AA1 was an independent prognostic factor for lung cancer patients. Conclusion Serum HSP90α and HSPAA1 in cancer tissues are elevated in patients with lung cancer, which should be used as an auxiliary diagnosis method for lung cancer. Meanwhile they should be used for therapeutic effect observation and survival status prediction.

Objective: To investigate the clinical features and pathogenic genes of a family with osteosclerosis. Methods: Six patients and six family members from a family in Jiangsu were tested for biochemical parameters, bone metabolic markers, bone mineral density, thoracolumbar anterior lateral slices, skull positive lateral radiographs, and pelvic plain films. Meanwhile, Sanger sequencing was performed to detect gene mutations of the proband and five other family members with high bone mass. The conformation of the mutational low-density lipoprotein receptor-related protein 5 (LRP5) protein was predicted by SWISS-MODEL. Results: Four adult patients (one male and three females) were tall, with mandibular enlargement and kyphosis in the center of the lower jaw, and none of the four had fractures. Their X ray examination revealed that the skull and long bone cortex was thickened, while the sella and mandible was enlarged. In addition, the absolute values of bone mineral density at each site of all patients were significantly higher as compared with the standard age- and sex-matched adults or adolescent mean reference values, with Z scores of L2-4, femoral neck and total hip being (6.31±4.03) SD, (6.56±2.36) SD, and (7.19±2.03) SD, respectively. The results of genetic sequencing revealed that all six patients carried a heterozygous mutation (c.331G>T; D111Y) in exon 2 of LRP5 gene, while other family members showed wild type (c.331G>G; D111D). Functional prediction indicated that this mutation was located at the amino acid terminal of exon 2 of LRP5 gene, which encodes the first β-helix-generating region of LRP5 protein. Conclusion The D111Y mutation in LRP5 gene leads to a clinical phenotype characterized by benign increased bone mineral density without increasing the risk of fracture. This mutation may further affect the downstream Wnt signaling pathway by altering the spatial structure of LRP5 protein, thereby promoting maturation and differentiation of osteoblasts and resulting in osteosclerosis.

Objective To analyze the changes in lung function and inflammatory indicators such as eosino-phils(EOS),Creola bodies and exhaled nitric oxide(FeNO),and to explore their value in early diagnosing asthma. Methods One hundred and thirty-five infants with high-risk asthma were selected as an observation group who were admitted to Department of Respiratory,Childrenˊs Hospital of Soochow University from April 2016 to August 2017 due to asthmatic bronchitis and asthmatic bronchopneumonia,and a total of 200 non -asthmatic and non -allergic healthy children who were screened and followed up at the Department of Cardiology in the same period were selected as a healthy control group for the measurement of moist respiratory lung function and FeNO. In the observation group,the bronchial dilation test(BDT),EOS and Creola bodies were performed simultaneously. Classification of lung function:23%≤time to peak ratio(TPTEF/Te)<28% had mild obstruction,15%≤TPTEF/Te<23% had moderate obstruc-tion,and TPTEF/Te<15% showed severe obstruction. Results The TPTEF/Te[17. 20%(8. 10%)],volume to peak ratio(VPEF/Ve)[21. 20%(6. 20%)],tidal volume per kilogram of body weight(VT/kg)[7. 80(3. 70)]and ratio of tidal expiratory flow 25% to peak tidal expiratory flow(25/PF)[0. 54(0. 20)]in the observation group were signifi-cantly lower than those in the healthy control group[22. 30%(9. 22%),27. 15%(7. 10%),8. 90(3. 17),0. 60 (0. 18)],and the differences were statistically significant(Z＝ -6. 81,-9. 35,-3. 16,-3. 52,all P<0. 05). BDT positive rate in the mild obstruction group was 20. 00%(3/15 cases),BDT positive rate in the moderate obstruction group was 26. 56%(17/64 cases),and BDT positive rate in the severe obstruction group was 48. 72%(19/39 cases). The higher the degree of obstruction,the higher the value of BDT positive diagnosis(F＝6. 353,P<0. 05). BDT of VPEF/Ve,25/PF and TPTEF/Te were consistent and statistically significant(Kappa＝0. 78,0. 49,all P<0. 001). Ti-dal expiratory flow 50% -remaining(TEF50% -r)[(117. 86 ± 42. 16)mL/s],tidal expiratory flow 25% -remai- ning(TEF25% -r)[(82. 82 ± 35. 44)mL/s]in the second wheezing group were higher than those in the first whee-zing group[(92. 81 ± 28. 40)mL/s,(65. 22 ± 24. 93)mL/s],and the differences were statistically significant( t＝3. 34,2. 77,all P <0. 05). There was no statistically significant difference in sputum EOS,FeNO and Creola body scores among wheezing children between the first and second groups(all P>0. 05). FeNO in the observation group [3. 80(5. 43)μg/L]was significantly lower than that in the healthy control group[9. 60(11. 3)μg/L],and the diffe-rence was statistically significant(Z＝14. 56,P<0. 05). Sputum EOS had a positive correlation with blood EOS and Creola bodies(r＝0. 20,0. 21,all P<0. 05);there was no correlation between lung function parameters and inflamma-tory indices(all P>0. 05). Conclusions In the acute phase of infant asthma attack,the pulmonary function presents different degrees of obstructive ventilation dysfunction,the higher the degree of obstruction,the higher the diagnostic value of BDT. The value of single FeNO measurement is limited,and continuous dynamic monitoring may be more mea-ningful in predicting the occurrence of asthma. Detection of Creola bodies may be helpful in predicting asthma.

For unresectable advanced hepatocellular carcinoma (HCC),besides sorafenib,alternative drugs and treatment modalities are required.Clinical studies of hepatic arterial infusion chemotherapy (HAIC),transcatheter arterial chemoembolization (TACE),and system chemotherapy have shown favorable efficacy and tolerance in advanced HCC patients.In addition,the potential efficacy of sorafenib combined with focal treatment is also an interesting issue.As more therapies become available,decision-making for treating advanced HCC becomes increasingly complex.In our opinion,diverse treatment modalities should be utilized for the best interest of patients.Based on predictive biomarkers,we should develop a precise patient stratification system to select suitable candidates for each treatment modality in future studies,as is useful for improving prognosis of patients with advanced HCC.

Objective To develop the clinical practice guidelines for the management of medication adherence to highly active antiretroviral therapy (HAART) in China. Methods The development methods included qualitative interview of 31 stakeholders, questionnaire survey of 423 PLHIV, adaptation of 30 clinical practice guidelines related to AIDS care, and overviews of reviews of 44 systematic reviews/Meta-analysis. Results 10 clinical practice guidelines and 10 systematic reviews/Meta-analysis were included. The clinical practice guidelines for the management of HAART were formed. Conclusions The formed clinical practice guidelines showed better applicability and higher general quality. It is recommended to use the guidelines in AIDS care.