ObjectiveTo investigate the therapeutic effect of sitravatinib on carbon tetrachloride （CCl₄）-induced liver fibrosis in mice. MethodsA total of 30 male C57BL/6J mice， aged 8 weeks， were randomly divided into control group， CCl₄ model group， and low- （5 mg/kg）， middle- （10 mg/kg）， and high-dose （20 mg/kg） sitravatinib groups. All mice except those in the control group were given intraperitoneal injection of CCl₄ for 4 consecutive weeks to induce liver fibrosis， and since the first day of modeling， the mice in the low-， middle-， and high-dose sitravatinib groups were given sitravatinib at the corresponding dose by gavage every day. The serum levels of total cholesterol （TC）， triglyceride （TG）， and alanine aminotransferase （ALT） were measured for the mice in each group； hepatic hydroxyproline content was measured； HE staining， Masson staining， and Sirius Red staining were used to observe liver histopathological changes； quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression levels of α-smooth muscle actin （α-SMA） and collagen type I alpha 1 （Col1a1） in liver tissue. The therapeutic effect of sitravatinib was assessed based on the above results. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the control group， the model group had significant increases in the levels of TC， TG， and ALT （all P<0.05）， and there were no significant differences in the levels of TC， TG， and ALT between the model group and the low-， middle-， and high-dose sitravatinib groups （all P>0.05）. Hepatic hydroxyproline content decreased after sitravatinib intervention， with a significant difference between the middle-/high-dose sitravatinib groups and the CCl₄ model group （both P<0.05）. Histopathological staining showed that the sitravatinib treatment groups had a reduction in collagen deposition， along with thinning and fragmentation of fibrous septa， and in the high-dose sitravatinib group， 4 mice had a fibrosis stage of S0—S1 and 2 mice had a fibrosis stage of S2—S3， suggesting a certain degree of alleviation of liver fibrosis degree compared with the CCl₄ model group （mainly S3—S4）. The measurement of related molecules showed that sitravatinib downregulated the mRNA and protein expression levels of α-SMA and Col1a1 （all P<0.05）. ConclusionSitravatinib can effectively alleviate CCl₄-induced liver fibrosis in mice， possibly by inhibiting hepatic stellate cell activation and collagen synthesis.

Objective:To investigate the effect of the antioxidant Tempol on the skin depigmentation of a vitiligo-like mouse model induced by the combination of the endogenous reactive oxygen species (ROS) producer AAPH and tyrosinase-related protein 2 (TRP2) -180 nonapeptide.Methods:A vitiligo-like skin depigmentation model was established by immunizing mice with injections of a TRP2-180 nonapeptide mixture into the foot pads twice and into the tails twice, with the injection interval being 1 week. After the first injection, 12 immune-induced mouse models of vitiligo were randomly divided into 4 groups (3 mice per group) : a model group, an AAPH group, a Tempol group, and a combined treatment group; additionally, 3 untreated mice injected with an ovalbumin (OVA) 257-264 peptide served as a sham control group. Mice in the AAPH group, the Tempol group, and the combined treatment group were subcutaneously injected with AAPH into the tails, intraperitoneally injected with Tempol, and received the above both treatments, respectively, while mice in the model group and the sham control group received phosphate-buffered saline injections into the tail and/or abdomen. Drug interventions were carried out 3 times per week for 3 consecutive weeks. Six weeks after the last immunization, mice were sacrificed. The area of depigmented macules on the tail was measured using a point-counting method, X-gal staining and double immunofluorescence staining were performed to assess the distribution and number of melanocytes, mast cells, and CD8 + T cells in depigmented macules on the tail. HaCaT cells were in vitro co-cultured with AAPH and/or Tempol, and a conventional culture group served as the control. Cellular ROS levels were measured by dichlorofluorescin diacetate labeling and flow cytometry; Western blot analysis was performed to determine the expression of matrix metalloproteinase 9 (MMP9) and stem cell factor (SCF) in cell lysates, and to detect soluble SCF levels in the culture supernatant. Comparisons among multiple groups were conducted using one-way analysis of variance, and multiple comparisons were performed using least significant difference- t test. Results:Depigmented macules were observed on the tails of mice in all groups except the sham control group. The area of depigmented macules was significantly larger in the AAPH group (7.27 ± 0.31 cm 2) than in the model group and combined treatment group (3.53 ± 0.21 cm 2, 4.07 ± 0.40 cm 2; t = 13.48, 11.56, respectively, both P < 0.001), while there was no significant difference between the Tempol group (3.30 ± 0.40 cm 2) and the model group ( P = 0.424). X-gal staining and double immunofluorescence staining showed that the number of melanocytes in the normal skin around the depigmented macules was significantly lower in the AAPH group and the combined treatment group than in the model group ( t = 6.31, 5.16, respectively, both P < 0.001), and no significant difference was observed between the AAPH group and the combined treatment group ( P = 0.516). The numbers of CD8 + T cells and mast cells were significantly higher in the AAPH group than in the model group and the combined treatment group (all P < 0.001). The numbers of the 3 types of cells mentioned above in the Tempol group did not differ from those in the model group (all P > 0.05). The ROS levels in HaCaT cells in the AAPH group were the highest, and significantly higher than those in the control group and the combined treatment group (both P < 0.001). Western blot analysis showed that the MMP9 level in the cell lysates and soluble SCF level in the culture supernatant were significantly higher in the AAPH group than in the control group and the combined treatment group (all P < 0.05) ; no significant difference was observed in the membrane-bound SCF level in cell lysates among the groups ( F = 0.06, P = 0.977) . Conclusion:The antioxidant Tempol could inhibit the formation of skin depigmented macules in vitiligo-like mouse models under AAPH-induced oxidative stress.

Objective:To investigate effects of rutin on ultraviolet irradiation (UVR) -induced human dermal fibroblast (FB) senescence and melanogenesis in mouse ear skin.Methods:The third- to fifth-passage FBs were divided into 4 groups: a blank control group, a UVR group, a rutin group, and a combined treatment group. In the UVR group, FBs were irradiated using an ultraviolet irradiator at a single dose of 0.6 J/cm 2 UVA combined with 0.03 J/cm 2 UVB once daily for 5 consecutive days; FBs in the rutin group were cultured in Dulbecco's modified Eagle medium containing 50 μmol/L rutin for 5 days; the combined treatment group received both UVR and the treatment with 50 μmol/L rutin for 5 days; the blank control group underwent no treatment. β-Galactosidase staining was performed to assess the senescence of FBs, real-time quantitative PCR (qPCR) to measure the telomere length in FBs, and Western blot analysis to detect the expression levels of stem cell factor (SCF) in FB cell lysates and culture supernatants. FB culture supernatants were collected from each group, and mixed with M254 medium at a ratio of 3∶1 to prepare conditioned medium, which was then used to treat PIG1 melanocytes for 24 hours. Western blot analysis was conducted to determine the tyrosinase (TYR) expression in PIG1 melanocytes in each group, while the 5-ethynyl-2′-deoxyuridine (EdU) incorporation assay was applied to assess the proliferative activity of PIG1 cells in each group. Ten Dct-LacZ transgenic mice were divided into a control group and a UVR group. For each mouse, 5% rutin-containing cream was topically applied to the right ear after UVR, while the left ear treated with the cream base alone served as a control. Skin biopsies were performed after 4 weeks, followed by X-gal staining and Avidin/fluorescein isothiocyanate (FITC) staining to count the numbers of melanocytes and mast cells in mouse ear skin. Results:In the UVR group, the number of senescent FBs (25.67 ± 2.89), relative protein expression levels of SCF (1.95 ± 0.22), and relative levels of SCF in the cell culture supernatant (1.52 ± 0.34) were all significantly higher than those in the blank control group (5.67 ± 1.56, 0.95 ± 0.11, 1.01 ± 0.31, respectively), while these indicators were significantly lower in the combined treatment group (12.00 ± 1.63, 1.32 ± 0.19, 1.15 ± 0.32, respectively) than in the UVR group (all P < 0.05). The relative telomere length in FBs was significantly shorter in the UVR group (0.49 ± 0.12) than in the blank control group (0.94 ± 0.11; LSD- t = 3.15, P = 0.021), but significantly longer in the combined treatment group (0.81 ± 0.13) than in the UVR group (LSD- t = 4.30, P = 0.034). After the treatment with FB conditioned medium, the relative expression level of TYR in PIG1 melanocytes and the number of EdU-positive cells were significantly higher in the UVR group (2.54 ± 0.21, 33.54 ± 3.21, respectively) than in the blank control group (0.97 ± 0.19, 21.45 ± 2.51, respectively; both P < 0.001), but significantly lower in the combined treatment group (1.63 ± 0.12, 18.54 ± 3.87, respectively) than in the UVR group (both P < 0.001). X-gal staining and Avidin/FITC staining showed that the numbers of melanocytes and mast cells in the mouse left ear skin in the UVR group (5.00 ± 1.22, 98.60 ± 8.47, respectively) were significantly higher than those in the mouse left ear skin in the control group (1.80 ± 0.45, 53.80 ± 5.76, respectively) and those in the mouse right ear skin treated with the rutin-containing cream in the UVR group (2.80 ± 0.45, 69.60 ± 8.89, respectively) (all P < 0.05) . Conclusion:Rutin may inhibit UVR-induced skin melanogenesis by suppressing the senescence of dermal FBs and paracrine secretion of SCF.

Objective:To investigate the effect of the antioxidant Tempol on the skin depigmentation of a vitiligo-like mouse model induced by the combination of the endogenous reactive oxygen species (ROS) producer AAPH and tyrosinase-related protein 2 (TRP2) -180 nonapeptide.Methods:A vitiligo-like skin depigmentation model was established by immunizing mice with injections of a TRP2-180 nonapeptide mixture into the foot pads twice and into the tails twice, with the injection interval being 1 week. After the first injection, 12 immune-induced mouse models of vitiligo were randomly divided into 4 groups (3 mice per group) : a model group, an AAPH group, a Tempol group, and a combined treatment group; additionally, 3 untreated mice injected with an ovalbumin (OVA) 257-264 peptide served as a sham control group. Mice in the AAPH group, the Tempol group, and the combined treatment group were subcutaneously injected with AAPH into the tails, intraperitoneally injected with Tempol, and received the above both treatments, respectively, while mice in the model group and the sham control group received phosphate-buffered saline injections into the tail and/or abdomen. Drug interventions were carried out 3 times per week for 3 consecutive weeks. Six weeks after the last immunization, mice were sacrificed. The area of depigmented macules on the tail was measured using a point-counting method, X-gal staining and double immunofluorescence staining were performed to assess the distribution and number of melanocytes, mast cells, and CD8 + T cells in depigmented macules on the tail. HaCaT cells were in vitro co-cultured with AAPH and/or Tempol, and a conventional culture group served as the control. Cellular ROS levels were measured by dichlorofluorescin diacetate labeling and flow cytometry; Western blot analysis was performed to determine the expression of matrix metalloproteinase 9 (MMP9) and stem cell factor (SCF) in cell lysates, and to detect soluble SCF levels in the culture supernatant. Comparisons among multiple groups were conducted using one-way analysis of variance, and multiple comparisons were performed using least significant difference- t test. Results:Depigmented macules were observed on the tails of mice in all groups except the sham control group. The area of depigmented macules was significantly larger in the AAPH group (7.27 ± 0.31 cm 2) than in the model group and combined treatment group (3.53 ± 0.21 cm 2, 4.07 ± 0.40 cm 2; t = 13.48, 11.56, respectively, both P < 0.001), while there was no significant difference between the Tempol group (3.30 ± 0.40 cm 2) and the model group ( P = 0.424). X-gal staining and double immunofluorescence staining showed that the number of melanocytes in the normal skin around the depigmented macules was significantly lower in the AAPH group and the combined treatment group than in the model group ( t = 6.31, 5.16, respectively, both P < 0.001), and no significant difference was observed between the AAPH group and the combined treatment group ( P = 0.516). The numbers of CD8 + T cells and mast cells were significantly higher in the AAPH group than in the model group and the combined treatment group (all P < 0.001). The numbers of the 3 types of cells mentioned above in the Tempol group did not differ from those in the model group (all P > 0.05). The ROS levels in HaCaT cells in the AAPH group were the highest, and significantly higher than those in the control group and the combined treatment group (both P < 0.001). Western blot analysis showed that the MMP9 level in the cell lysates and soluble SCF level in the culture supernatant were significantly higher in the AAPH group than in the control group and the combined treatment group (all P < 0.05) ; no significant difference was observed in the membrane-bound SCF level in cell lysates among the groups ( F = 0.06, P = 0.977) . Conclusion:The antioxidant Tempol could inhibit the formation of skin depigmented macules in vitiligo-like mouse models under AAPH-induced oxidative stress.

Objective:To investigate effects of rutin on ultraviolet irradiation (UVR) -induced human dermal fibroblast (FB) senescence and melanogenesis in mouse ear skin.Methods:The third- to fifth-passage FBs were divided into 4 groups: a blank control group, a UVR group, a rutin group, and a combined treatment group. In the UVR group, FBs were irradiated using an ultraviolet irradiator at a single dose of 0.6 J/cm 2 UVA combined with 0.03 J/cm 2 UVB once daily for 5 consecutive days; FBs in the rutin group were cultured in Dulbecco's modified Eagle medium containing 50 μmol/L rutin for 5 days; the combined treatment group received both UVR and the treatment with 50 μmol/L rutin for 5 days; the blank control group underwent no treatment. β-Galactosidase staining was performed to assess the senescence of FBs, real-time quantitative PCR (qPCR) to measure the telomere length in FBs, and Western blot analysis to detect the expression levels of stem cell factor (SCF) in FB cell lysates and culture supernatants. FB culture supernatants were collected from each group, and mixed with M254 medium at a ratio of 3∶1 to prepare conditioned medium, which was then used to treat PIG1 melanocytes for 24 hours. Western blot analysis was conducted to determine the tyrosinase (TYR) expression in PIG1 melanocytes in each group, while the 5-ethynyl-2′-deoxyuridine (EdU) incorporation assay was applied to assess the proliferative activity of PIG1 cells in each group. Ten Dct-LacZ transgenic mice were divided into a control group and a UVR group. For each mouse, 5% rutin-containing cream was topically applied to the right ear after UVR, while the left ear treated with the cream base alone served as a control. Skin biopsies were performed after 4 weeks, followed by X-gal staining and Avidin/fluorescein isothiocyanate (FITC) staining to count the numbers of melanocytes and mast cells in mouse ear skin. Results:In the UVR group, the number of senescent FBs (25.67 ± 2.89), relative protein expression levels of SCF (1.95 ± 0.22), and relative levels of SCF in the cell culture supernatant (1.52 ± 0.34) were all significantly higher than those in the blank control group (5.67 ± 1.56, 0.95 ± 0.11, 1.01 ± 0.31, respectively), while these indicators were significantly lower in the combined treatment group (12.00 ± 1.63, 1.32 ± 0.19, 1.15 ± 0.32, respectively) than in the UVR group (all P < 0.05). The relative telomere length in FBs was significantly shorter in the UVR group (0.49 ± 0.12) than in the blank control group (0.94 ± 0.11; LSD- t = 3.15, P = 0.021), but significantly longer in the combined treatment group (0.81 ± 0.13) than in the UVR group (LSD- t = 4.30, P = 0.034). After the treatment with FB conditioned medium, the relative expression level of TYR in PIG1 melanocytes and the number of EdU-positive cells were significantly higher in the UVR group (2.54 ± 0.21, 33.54 ± 3.21, respectively) than in the blank control group (0.97 ± 0.19, 21.45 ± 2.51, respectively; both P < 0.001), but significantly lower in the combined treatment group (1.63 ± 0.12, 18.54 ± 3.87, respectively) than in the UVR group (both P < 0.001). X-gal staining and Avidin/FITC staining showed that the numbers of melanocytes and mast cells in the mouse left ear skin in the UVR group (5.00 ± 1.22, 98.60 ± 8.47, respectively) were significantly higher than those in the mouse left ear skin in the control group (1.80 ± 0.45, 53.80 ± 5.76, respectively) and those in the mouse right ear skin treated with the rutin-containing cream in the UVR group (2.80 ± 0.45, 69.60 ± 8.89, respectively) (all P < 0.05) . Conclusion:Rutin may inhibit UVR-induced skin melanogenesis by suppressing the senescence of dermal FBs and paracrine secretion of SCF.

Objective The objective of this study was to analyze the long-term change trend of gastric cancer incidence in China from 1990 to 2021,and to provide scientific suggestion for the prevention and control of gastric cancer in China.Methods The incidence burden data of gastric cancer from 1990 to 2021 were obtained from the Global Burden of Disease(GBD)2021 database.The age-period-cohort model was to evaluate its independent effects,and the Nordpred model was used to predict its incidence trend from 2022 to 2031.Results The number and incidence of gastric cancer from 1990 to 2021 in China showed an upward trend,while the age-standardized incidence showed a decreasing trend.The results of age-period-cohort model showed that in the same birth cohort,the incidence of gastric cancer in China increased with age,the incidence of the total population increases from 0.91/100,000 to 233.37/100,000 in total population,the incidence of men increases from 0.91/100,000 to 508.90/100,000,and the incidence of women increases from 1.04/100,000 to 115.86/100,000.During the period from 1992-1996 to 2017-2021,the relative risk(RR)of gastric cancer incidence showed a decreasing trend with the passage of time.The RR of the total population period decreased from 1.10 to 0.72,with males decreasing from 1.06 to 0.75 and females decreasing from 1.19 to 0.65.The later the birth cohort through-out the birth cohort years,the lower the risk of onset;The RR of gastric cancer incidence in the total population decreased from 1.60 to 0.34,from 1.36 to 0.40 for males and from 2.23 to 0.22 for females.The incidence prediction results showed that by 2031,the in-cidence of gastric cancer would increase to 470,429 cases,including 323,399 cases for men and 147,029 cases for women.The stand-ardized incidence of the total population would decrease to 23.80/100,000 cases,35.13/100,000 cases for men and 24.17/100,000 cases for women.Conclusion The prevention and treatment measures of gastric cancer from 1990 to 2021 have achieved certain re-sults in reducing the risk of gastric cancer,but the incidence of gastric cancer is still serious,and it is necessary to focus on strengthe-ning the intervention for men over 50 years old.

Objective The objective of this study was to analyze the long-term change trend of gastric cancer incidence in China from 1990 to 2021,and to provide scientific suggestion for the prevention and control of gastric cancer in China.Methods The incidence burden data of gastric cancer from 1990 to 2021 were obtained from the Global Burden of Disease(GBD)2021 database.The age-period-cohort model was to evaluate its independent effects,and the Nordpred model was used to predict its incidence trend from 2022 to 2031.Results The number and incidence of gastric cancer from 1990 to 2021 in China showed an upward trend,while the age-standardized incidence showed a decreasing trend.The results of age-period-cohort model showed that in the same birth cohort,the incidence of gastric cancer in China increased with age,the incidence of the total population increases from 0.91/100,000 to 233.37/100,000 in total population,the incidence of men increases from 0.91/100,000 to 508.90/100,000,and the incidence of women increases from 1.04/100,000 to 115.86/100,000.During the period from 1992-1996 to 2017-2021,the relative risk(RR)of gastric cancer incidence showed a decreasing trend with the passage of time.The RR of the total population period decreased from 1.10 to 0.72,with males decreasing from 1.06 to 0.75 and females decreasing from 1.19 to 0.65.The later the birth cohort through-out the birth cohort years,the lower the risk of onset;The RR of gastric cancer incidence in the total population decreased from 1.60 to 0.34,from 1.36 to 0.40 for males and from 2.23 to 0.22 for females.The incidence prediction results showed that by 2031,the in-cidence of gastric cancer would increase to 470,429 cases,including 323,399 cases for men and 147,029 cases for women.The stand-ardized incidence of the total population would decrease to 23.80/100,000 cases,35.13/100,000 cases for men and 24.17/100,000 cases for women.Conclusion The prevention and treatment measures of gastric cancer from 1990 to 2021 have achieved certain re-sults in reducing the risk of gastric cancer,but the incidence of gastric cancer is still serious,and it is necessary to focus on strengthe-ning the intervention for men over 50 years old.

Objective To explore the correlation of single nucleotide polymorphism ( SNP ) in Nrf2 gene with acute mountain sickness ( AMS) among Han populations in China .Methods As a nested case-control study , 603 Chinese Han young men who had been quickly exposed to 3700 m were adopted and divided into case group and control group ( 369 vs 234,respectively) by Lake Louise Self-assessment Scoring System(LLSS).The Sequenom Mass Array system was used to detect the SNPs of rs10497511 and rs2364722 in Nrf2 gene.Results Alleles of rs10497511 and rs2364722 were respec-tively detected in both case and control groups , which were T-C and A-G.Statistically significant difference was not found in allele frequencies ( P>0.05 ) .Further analysis showed that there was still no significant difference between the codomi -nant, dominant and recessive genotype models (P>0.05).Conclusion rs10497511 and rs2364722 of Nrf2 gene may not be related to susceptibility to AMS in Chinese Han populations .