Objective To investigate the effect of temperature on cell proliferation and osteogenic differentiation inhibition of preosteoblast induced by hydrogen peroxide(H2 O2).Methods The MC3T3-E1 cells in the logarithmic phase were randomly divided into 0,450,500,550,600,650 μmol·L-1 H2O2 intervention groups and incubated with 0,450,500,550,600,650 μmol·L-1 H2O2 for 2 h,respectively.Other MC3T3-E1 cells in the logarithmic phase were selected and randomly divided into the control group,model group,low-temperature group,and high-temperature group.Cells in the control group were cul-tured in an incubator with 5％CO2 for 24 h at 37 ℃;cells in the model group were incubated with H2O2 for 2 h and cultured in an incubator with 5％CO2 for 24 h at 37 ℃;cells in the low-temperature group were incubated with H2O2 for 2 h and cultured in an incubator with 5％CO2 for 24 h at 32 ℃;cells in the high-temperature group were incubated with H2O2 for 2 h and cultured in an incubator with 5％CO2 for 24 h at 40 ℃.The cell proliferation in all groups was detected by cell counting kit-8.The expression levels of Runt-related transcription factor 2(RUNX2),osteopontin(OPN)and osteocalcin(OC)mRNA were detected by real-time fluorescence quantitave polymerase chain reaction;and the expression levels of RUNX2,OPN and OC protein were detected by Western blot.Results There was no statistically significant difference in cell proliferation among the 0,450 and 500 μmol·L-1 H2O2 intervention groups(P＞0.05);the cell proliferation rate in the 550,600 and 650 μmol·L-1 H2O2 intervention groups was significantly lower than that in the 0,450 and 500 μmol·L-1 H2O2 intervention groups,showing a significant decrease in cell proliferation with the increase of H2O2 concentrations(P＜0.05).In order to ensure that there were enough cells to perform the following experiments,550 μmol·L-1 H2 O2 was chosen.The cell proliferation rate in the model group and the low-temperature group was significantly lower than that in the control group and high-temperature group(P＜0.05);there was no significant difference in the cell proliferation rate between the control group and high-temperature group(P＞0.05).The relative expression of RUNX2 mRNA in the model group and high-temperature group were significantly higher than that in the control group and low-temperature group(P＜0.05);the relative expression of RUNX2 mRNA in the low-temperature group was significantly lower than that in the control group(P＜0.05);there was no significant difference in the relative expression of RUNX2 mRNA between the model group and high-temperature group(P＞0.05).The relative expression of OPN mRNA in the model group,low-temperature group and high-temperature group was significantly higher than that in the control group(P＜0.05);the relative expression of OPN mRNA in the low-temperature group and high-temperature group was significantly higher than that in the model group(P＜0.05);the relative expression of OPN mRNA in the low-tem-perature group was significantly higher than that in the high-temperature group(P＜0.05).The relative expression of OC mRNA in the model group,low-temperature group and high-temperature group was significantly than that in the control group(P＜0.05);the relative expression of OC mRNA in the low-temperature group and high-temperature group was significantly higher than that in the model group(P＜0.05);there was no significant difference in the relative expression of OC mRNA between the low-temperature group and high-temperature group(P＞0.05).The relative expressions of RUNX2,OPN and OC protein the model group,low-temperature group and high-temperature group were significantly lower than those in the control group(P＜0.05);the relative expressions of RUNX2 and OPN protein in the low-temperature group were significantly lower than those in the model group and high-temperature group(P＜0.05);the relative expression of OC protein was significantly lower than that in the high-temperature group(P＜0.05);and there was no siqnificantly difference in the relatiwe experesson of OC protein between the low-temperature group and model group(P＞0.05);the relative expressions of RUNX2,OPN and OC protein in the high-temperature group were significantly higher than those in the model group(P＜0.05).Conclusion The inhibitory effects of H2O2 on cell proliferation and osteogenic differentiation are observed in MC3T3-E1 cells;low-tempera-ture incubation can enhance the inhibition of H2O2 on cell proliferation and osteogenic differentiation in MC3T3-E1 cells,while high-temperature incubation can relieve its inhibitory effect on cell proliferation and osteogenic differentiation.RUNX2,OPN and OC protein might play an important role in cell proliferation and osteogenic differentiation mediated by temperature.

Tuberous sclerosis complex(TSC)is a rare genetic disease that can lead to benign dysplasia in multiple organs such as the skin, brain, eyes, oral cavity, heart, lungs, kidneys, liver, and bones. Its main symptoms include epilepsy, intellectual disabilities, skin depigmentation, and facial angiofibromas, whilst incidence is approximately 1 in 10 000 to 1 in 6000 newborns. This case presents a middle-aged woman who initially manifested with epilepsy and nodular depigmentation. Later, she developed a lower abdominal mass, elevated creatinine, and severe anemia. Based on clinical features and whole exome sequencing, the primary diagnosis was confirmed as TSC. Laboratory and imaging examinations revealed that the lower abdominal mass originated from the uterus. CT-guided biopsy pathology and surgical pathology suggested a combination of leiomyoma and abscess. With the involvement of multiple organs and various complications beyond the main diagnosis, the diagnostic and therapeutic process for this patient highlights the importance of rigorous clinical thinking and multidisciplinary collaboration in the diagnosis and treatment of rare and challenging diseases.

ObjectiveTo investigate the pollution level of deoxynivalenol （DON） in wheat flour and its products sold in Shanghai， and to assess the health risks of DON exposure for residents in Shanghai who ingested DON from wheat flour and its products. MethodsRisk monitoring data of DON in wheat flour and its products sold in Shanghai from 2017 to 2021 were combined with the consumption data of wheat flour and its products by Shanghai residents. A probabilistic assessment method was used to assess dietary exposure of DON in wheat flour and its products. ResultsThe overall detection rate of DON in wheat flour and its products was 77.3% （1 041/1 347）， with a mean concentration of 226.3 μg·kg^-1， P₅₀ of 130.0 μg·kg^-1 and a maximum value of 3 080.0 μg·kg^-1. The mean daily exposure and 95th percentile daily exposure （by body weight） of DON from wheat flour and its products in Shanghai residents were 0.279 μg·kg^-1 and 1.146 μg·kg， accounting for 27.9% and 114.6% of the daily tolerable intake of DON TDI， 1 μg·kg， respectively. The probability assessment results indicated that 6.1% of the whole population in Shanghai had DON exposure exceeding the TDI value. Among them， 12.8% of the population aged 6 years old and below， 16.4% of the population aged between 7 and 17 years old， 3.9% of the population aged between 18 and 59 years old and 3.2% of the population aged 60 years old and above exceeded the TDI value for daily DON exposure through wheat flour and its products. ConclusionCertain populations in Shanghai may face certain health risks from daily DON intake wheat flour and its products. Special attention should be paid to the health risk of daily DON exposure through wheat flour and its products for individuals age below 18 years old .

ObjectiveTo assess the health risk of dietary exposure to nonylphenol in infants aged 0-36 months through infant formula in Shanghai. MethodsA monitoring of nonylphenol pollution in infant formula was conducted in 2022. A total of 90 samples were obtained from maternal and infant stores， supermarkets， and online stores in Shanghai. Based on the daily consumption data of infant formula， a point assessment method was used to assess the dietary exposure to nonylphenol in infant formula. ResultsThe prevalence of nonylphenol in infant formula retailed in Shanghai was 95.6% （86/90）. The amount of nonylphenol varied from non-detected to 22.70 μg·kg^-1， with the mean value of 8.47 μg·kg^-1 and the P₅₀ value of 7.77 μg·kg^-1. The mean daily nonylphenol exposure （estimated by body weight） from infant formula in infants aged 0-6 months， 7-12 months and 13-36 months in Shanghai was 0.091， 0.068 and 0.054 μg·kg^-1， respectively； furthermore， the P₉₅ value of daily exposure （by body weight） was 0.228， 0.152 and 0.119 μg·kg^-1， respectively. These amounts were much lower than the tolerable daily intake （TDI） of nonylphenol （by body weight 5 μg·kg^-1）. ConclusionThe health risk of daliy nonylphenol intake from infant formula remains low among infants aged 0-36 months in Shanghai.

Objective:To evaluate the clinical efficacy and safety of a skin care ointment containing oligomeric maltose X in the adjuvant treatment of eczema-related pruritus.Methods:A multicenter, randomized, double-blind, vehicle-controlled clinical study was conducted. From March to September 2021, outpatients with mild to moderate eczema were collected from departments of dermatology of 4 hospitals, including Beijing Friendship Hospital, Hebei Traditional Chinese Medical Hospital, the Third People′s Hospital of Hubei Province, and Taizhou Central Hospital in Zhejiang Province. The patients were randomly divided into two groups by using a random number table: observation group topically treated with a skin care ointment containing oligomeric maltose X, and vehicle control group topically treated with an ointment vehicle. The ointments were applied during the attacks of itching for 14 consecutive days. Visits were scheduled before, 7, and 14 days after the start of the adjuvant treatment. The efficacy was evaluated according to the eczema area and severity index (EASI) and visual analog scale (VAS) , and adverse events were recorded. The efficacy and safety analyses were conducted by using chi-square test and t test. Results:Totally, 232 patients with eczema were enrolled, including 90 males and 142 females, with the age being 40.13 ± 13.36 years; 156 patients were in the observation group, and 76 in the vehicle control group. Before the adjuvant treatment, there were no significant differences in EASI (2.07 ± 2.24 points vs. 2.29 ± 2.28 points) or VAS (6.22 ± 1.78 points vs. 6.20 ± 1.79 points) scores between the observation group and vehicle control group ( t = -0.70, 0.06, P = 0.486, 0.955, respectively) . After one-day treatment, the VAS scores significantly decreased compared with the baseline scores in the two groups ( P < 0.001, P = 0.003, respectively) . After 14-day treatment, the VAS score was significantly lower in the observation group (2.67 points) than in the vehicle control group (3.35 points; t = -2.28, P = 0.024) . After 7- and 14-day treatment, the EASI scores significantly decreased compared with the baseline scores in both the two groups (all P < 0.001) , but there were no significant differences between the two groups ( P = 0.853, 0.731) . No adjuvant treatment-related adverse events were recorded in either of the two groups. Conclusion:The skin care ointment containing oligomeric maltose X is safe and effective in the adjuvant treatment of eczema-related pruritus, and can be applied to clinical practice.

ObjectiveTo investigate chlorate contamination level in infant formula sold in Shanghai， and to evaluate the dietary exposure risk to infants in Shanghai. MethodsWith the risk monitoring data of chlorate in infant formula sold in Shanghai in 2020， combined with the dietary consumption data of infants， the dietary exposure of chlorate in infant formula was assessed via the point assessment method. ResultsIn 2020， the overall detection rate of chlorate in 120 infant formula samples was 98.3% （118/120）， the mean content was 124.5 μg⋅kg^-1， the 50 percentile value was 64.6 μg⋅kg^-1， and the maximum value was 1 475.0 μg⋅kg^-1. The mean and 95 percentile value of daily chlorate intake from infant formula for infants aged 0‒36 months in Shanghai were 1.10 and 1.84 μg⋅kg^-1， accounting for 36.7% and 61.3% of the tolerable daily intake （TDI） of chlorate （3μg⋅kg^-1）， respectively. The mean， 50 percentile value and 95 percentile value of daily chlorate exposure of infants in different month-age groups （0‒6 months， 6‒12 months， 12‒36 months） through infant formula were lower than the TDI value. ConclusionThe health risk of daily chlorate intake from infant formula for infants and young children aged 0‒36 months in Shanghai is at an acceptable level.

Objective:To analyze the application value of sakubatril valsartan in the treatment of chronic heart failure (CHF) based on cardiopulmonary test system.Methods:One hundred and thirty-five CHF patients admitted to the Affiliated Hospital of Jining Medical Collegefrom January 2019 to August 2020 were divided into the observation group (67 cases) and the control group (68cases) by random number table method. Both groups were treated with bisoprolol. The observation group was treated with the combination of sakubatril valsartan, and the control group was treated with the combination of benapril. The efficacy and cardiac function indicators of the two groups were compared. The cardiopulmonary exercise test system was used to measure the patient′s maximum exercise time (Tmax), maximum exercise Watt (Wmax), peak volume oxygen (Peak VO 2) and volume of anaerobic threshold oxygen (VO 2AT), and the incidence of adverse reactions were calculated. Results:The total effective rate in the observation group was higher than that in the control group: 92.54% (62/67) vs. 77.94%(53/68), the difference was statistically significant ( χ2 = 5.70, P<0.05). After the treatment, the levels of N-terminal pro-brain natriuretic peptide (NT-proBNP) and soluble ST2 (sST2) in the observation group were lower than those in the control group: (2 000.47 ± 517.85) ng/L vs. (2 777.39 ± 812.49) ng/L, (0.33 ± 0.10) μg/L vs. (0.37 ± 0.09) μg/L, and the left ventricular ejection fraction (LVEF) was higher than that in the control group: (8.12 ± 6.44)% vs. (41.93 ± 6.73)%, the differences were statistically significant ( P<0.05). After the treatment, the left ventricular end-diastolic diameter (LVEDd), left ventricular end-systolic diameter (LVEDs), left ventricular mass index (LVMI), left atrial volume index (LAVI) in the observation group were lower than those in the control group: (55.47 ± 6.93) mm vs. (62.00 ± 7.18) mm, (37.14 ± 6.36) mm vs. (41.35 ± 6.43) mm, (136.76 ± 7.13) mg/m 2 vs. (140.98 ± 7.47) mg/m 2, (28.23 ± 2.59) ml/m 2 vs. (31.98 ± 2.17) ml/m 2; the Tmax, Wmax, PeakVO 2 and VO 2AT in the observation group were higher than those in the control group: (619.08 ± 65.36) s vs. (58.70 ± 52.44) s, (142.96 ± 16.05) W vs. (124.19 ± 13.38) W, (20.00 ± 5.74) ml/(min·kg) vs. (18.13 ± 3.58) ml/(min·kg), (13.89 ± 3.69) ml/(min·kg) vs. (11.23 ± 2.36) ml/(min·kg), the differences were statistically significant ( P<0.05). However, there was no statistically significant in the incidence of adverse reactions between the two groups ( P>0.05). Conclusions:Sakubatril valsartan in the treatment of CHF can not only optimize the efficacy and improve cardiac function, but also benefit cardiac exercise rehabilitation of patients, and not increase the safety risk.

Objective:To compare the efficacy of sakubatril valsartan and valsartan in the treatment of patients with chronic cardiac insufficiency and the influence on zinc finger protein A20 and nuclear factor-κB (NF-κB) in peripheral bloodmononuclear cells (PBMCs).Methods:Ninety-senven patients with chronic cardiac insufficiency admitted to the Affiliated Hospital of Jining Medical College from February 2019 to January 2020 were continuously selected and randomly divided into the control group (48 cases) and the observation group (49 cases). Both groups received routine anti-heart failure according to the guidelines. The control group added with valsartan and the observation group added with sakubatril valsartan treatment. Before the treatment and after 3 months of treatment, the changes of cardiac function indexes and the changes of inflammatory markers such as hypersensitive C-reactive protein (hs-CRP), tumor necrosis factor-α (TNF-α), matrix metalloproteinase 9 (MMP-9), and N-terminal pro B-type natriuretic peptide (NT-proBNP) were compared. PBMCs was extracted to detect zinc finger protein A20 and NF-κB levels. The incidence of adverse reactions in the two groups was recorded, and the relationship between zinc finger proteins A20, NF-κB and the myocardial injury marker NT-proBNP were analyzed.Results:After 3 months of treatment, the changes of cardiac function indexes in the observation group were better than those in the control group and the levels of hs-CRP, TNF-α, MMP-9, NT-proBNP in the observation group were lower than those in the control group: (1.96 ± 0.57) mg/L vs. (2.87 ± 0.79) mg/L, (7.11 ± 1.46) μg/L vs. (8.24 ± 1.57) μg/L, (110.14 ± 10.63) μg/L vs. (129.52 ± 17.96) μg/L, (716.91 ± 105.78) ng/L vs. (965.25 ± 97.41) ng/L, there were statistical differences ( P<0.05). After 3 months of treatment, the levels of finger protein A20, NF-κB in the observation group were lower than those in the control group: (3.57 ± 1.13) % vs. (4.41 ± 1.32) %, (29.87 ± 6.58) ng/L vs. (35.71 ± 10.02) ng/L, there were statistical differences ( P<0.05). Finger protein A20 and NF-κB in patients with chronic cardiac insufficiency were positively correlated with NT-proBNP ( r = 0.487, 0.738, P<0.01). Conclusions:On the basis of conventional treatment, compared with valsartan, the addition of sakubatril valsartan, can improve the cardiac function of patients with chronic cardiac insufficiency, reduce the body′s inflammatory response, reduce the expression of myocardial injury marker NT-proBNP, inhibit the activation of PBMCs NF-κB, and reduce the level offinger protein A20.

Background::Gut-resident macrophages (gMacs) supplemented by monocytes-to-gMacs differentiation play a critical role in maintaining intestinal homeostasis. Activating transcription factor 4 (ATF4) is involved in immune cell differentiation. We therefore set out to investigate the role of ATF4-regulated monocytes-to-gMacs differentiation in sepsis-induced intestinal injury.Methods::Sepsis was induced in C57BL/6 wild type (WT) mice and Atf4-knockdown ( Atf4+/-) mice by cecal ligation and puncture or administration of lipopolysaccharide (LPS). Colon, peripheral blood mononuclear cells, sera, lung, liver, and mesenteric lymph nodes were collected for flow cytometry, hematoxylin and eosin staining, immunohistochemistry, quantitative reverse transcription polymerase chain reaction, and enzyme-linked immunosorbent assay, respectively. Results::CD64, CD11b, Ly6C, major histocompatibility complex-II (MHC-II), CX3CR1, Ly6G, and SSC were identified as optimal primary markers for detecting the process of monocytes-to-gMacs differentiation in the colon of WT mice. Monocytes-to-gMacs differentiation was impaired in the colon during sepsis and was associated with decreased expression of ATF4 in P1 (Ly6C hi monocytes), the precursor cells of gMacs. Atf4 knockdown exacerbated the impairment of monocytes-to-gMacs differentiation in response to LPS, resulting in a significant reduction of gMacs in the colon. Furthermore, compared with WT mice, Atf4+/- mice exhibited higher pathology scores, increased expression of inflammatory factor genes ( TNF-α, IL-1β), suppressed expression of CD31 and vascular endothelial-cadherin in the colon, and increased translocation of intestinal bacteria to lymph nodes and lungs following exposure to LPS. However, the aggravation of sepsis-induced intestinal injury resulting from Atf4 knockdown was not caused by the enhanced inflammatory effect of Ly6C hi monocytes and gMacs. Conclusion::ATF4, as a novel regulator of monocytes-to-gMacs differentiation, plays a critical role in protecting mice against sepsis-induced intestinal injury, suggesting that ATF4 might be a potential therapeutic target for sepsis treatment.

Objective:To investigate the effect and mechanism of 6-formylindolo[3,2-b]carbazole (FICZ) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice.Methods:Male C57BL/6J mice aged 8-12 weeks were divided into 4 groups with 8 mice in each group, according to the method of simple random sampling. Sepsis-induced ALI mice model was established by intraperitoneal injection of LPS 5 mg/kg (LPS group), and phosphate buffer saline (PBS) control group (PBS group) was injected with equal volume of PBS. The LPS+FICZ group was intervened by intraperitoneal injection of 1 μg FICZ 1 hour after LPS stimuli, while the FICZ control group (FICZ group) was given the same amount of FICZ 1 hour after intraperitoneal injection of PBS. Serum and lung tissue were collected 24 hours after LPS stimuli, and the pathological changes of lung tissue were analyzed by hematoxylin-eosin (HE) staining and wet/dry weight (W/D) ratio of lung tissue. The concentrations of inflammatory factors in serum and lung tissue were detected by enzyme linked immunosorbent assay (ELISA). The expression levels of endoplasmic reticulum stress signaling pathway related molecules were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) and Western blotting.Results:Compared with PBS group, inflammatory cell infiltration, alveolar collapse and obvious alveolar exudative lesions had increased, lung tissue W/D ratio was significantly increased, serum interleukin-6 (IL-6) level, lung tissue IL-6 mRNA expression, and the mRNA expressions of glucose-regulated protein 78 (GRP78), protein kinase R-like endoplasmic reticulum kinase (PERK), CCAAT/EBP homologous protein (CHOP), and the protein expressions of GRP78, PERK, activating transcription factor 6 (ATF6), CHOP in lung tissue were significantly increased in LPS group. However, the indexes of FICZ group were not affected. Compared with LPS group, LPS+FICZ group had less inflammatory cell infiltration, relatively intact alveolar structure. Lung W/D weight ratio in LPS+FICZ group was significantly decreased (5.38±0.10 vs. 6.60±0.30, P < 0.01), so as serum IL-6 (ng/L: 15.55±3.77 vs. 32.22±3.84) and lung IL-6 mRNA expression (2 -ΔΔCt: 0.79±0.21 vs. 6.89±0.92, both P < 0.01). The mRNA expressions of GRP78, PERK and CHOP were also significantly decreased [GRP78 mRNA (2 -ΔΔCt): 1.90±0.16 vs. 7.55±1.29, PERK mRNA (2 -ΔΔCt): 1.68±0.20 vs. 4.54±0.89, CHOP mRNA (2 -ΔΔCt): 1.13±0.24 vs. 4.44±1.13, all P < 0.05], and the protein expressions of GRP78, PERK, ATF6 and CHOP were significantly decreased (GRP78/GAPDH: 0.59±0.02 vs. 0.77±0.01, PERK/GAPDH: 0.48±0.03 vs. 1.04±0.05, ATF6/GAPDH: 0.51±0.03 vs. 0.65±0.01, CHOP/GAPDH: 0.91±0.05 vs. 1.11±0.07, all P < 0.05). Conclusion:FICZ protects LPS-induced ALI possibly via suppressing endoplasmic reticulum stress and reducing IL-6 expression in blood and lung tissue.