ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus， in order to obtain the optimal fermentation conditions and flora structure， and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2（ITS2） high-throughput sequencing， combined with fungal community diversity analysis and fungal community structure analysis， were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7， 14， 21 d（numbered Y1-Y3 for the former， and numbered F1-F3 for the latter）， respectively. At the same time， the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， combined with principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and compound retention time， parent ions， characteristic fragment ions and other information， the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method， while in the non-pressure-shelf natural fermentation method， there was a significant difference with the fermentation process， and at the genus level， the dominant genus of samples Y1， Y2， Y3 and F2 was Aspergillus， while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable， and the microbial community structure was more stable， which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis， including 70 flavonoids， 38 coumarins， 10 alkaloids， 34 organic acids and 3 other compounds. After fermentation， two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition， multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods， mainly including flavonoids， organic acids and coumarins. Comprehensively， the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable， the species richness was high and the overall content of differential compounds was high， which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation， the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds， and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus， as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.

ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus， in order to obtain the optimal fermentation conditions and flora structure， and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2（ITS2） high-throughput sequencing， combined with fungal community diversity analysis and fungal community structure analysis， were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7， 14， 21 d（numbered Y1-Y3 for the former， and numbered F1-F3 for the latter）， respectively. At the same time， the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， combined with principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and compound retention time， parent ions， characteristic fragment ions and other information， the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method， while in the non-pressure-shelf natural fermentation method， there was a significant difference with the fermentation process， and at the genus level， the dominant genus of samples Y1， Y2， Y3 and F2 was Aspergillus， while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable， and the microbial community structure was more stable， which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis， including 70 flavonoids， 38 coumarins， 10 alkaloids， 34 organic acids and 3 other compounds. After fermentation， two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition， multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods， mainly including flavonoids， organic acids and coumarins. Comprehensively， the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable， the species richness was high and the overall content of differential compounds was high， which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation， the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds， and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus， as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.

AIM:This study explores whether curcumin(Cur)promotes mitophagy to attenuate nonalcoholic steatohepatitis(NASH)in mice,as well as the possible molecular mechanisms involved.METHODS:A high-fat and high-cholesterol diet was used to replicate the NASH mouse model.Thirty-two male C57BL/6J mice were randomly divided into normal control(NC)group,high-fat and high-cholesterol model(M)group,M+low-dose Cur(Cur-L)group,and M+high-dose Cur(Cur-H)group,with 8 mice in each group.The weight of 8 mice in each group was recorded weekly.After feeding for 18 weeks,the serum and liver of mice were collected.Serum levels of total cholesterol(TC),triglyceride(TG),low-density lipoprotein(LDL-C),alanine aminotransferase(ALT),aspartate aminotransferase(AST),and tumor necrosis factors-α(TNF-α)were measured.Liver index was calculated,and steatosis,inflammation,and fibrosis of the liver were observed by HE and Masson staining.Western blot analysis was performed to detect the protein expression of mi-tophagy-related protein,TNF-α and α-SMA in the liver.(2)HepG2 cells were treated with oleic acid and cholesterol to replicate the hepatocyte injury model,which was divided into NC group,Cur group,M group,and M+Cur group.Small interfering RNA for PTEN-induced kinase 1(PINK1)knockdown was used to explore the relationship between PINK1-me-diated mitophagy and NASH.Compound C(CC)was used to inhibit AMP-activated protein kinase(AMPK)to explore the effect of the AMPK/silent information regulator 1(Sirt1)pathway on mitophagy.The lipid droplets of HepG2 cells were ob-served by oil red O staining,and the levels of TC,TG,LDL-C,ALT,and AST in cell suspension were detected.RE-SULTS:(1)Compared with M group,treatment with Cur significantly reduced the body weight,liver coefficient,and se-rum levels of TC,TG,LDL-C,ALT,AST,and TNF-α in NASH mice,while the steatosis and fibrosis in the liver were improved(P<0.05).(2)Different concentrations of Cur could increase or decrease the expression of mitophagy-related proteins in HepG2 cells in a concentration gradient.Compared with the M group,Cur reduced lipid droplets and de-creased TC,TG,LDL-C,ALT,and AST levels(P<0.05).(3)Compared with the NC group,the expression levels of mi-tophagy-related proteins in the liver of mice in the M group decreased,and the expression levels of TNF-α and α-SMA pro-teins increased.Different concentrations of Cur intervention promoted the increase of mitophagy-related proteins and the decrease of TNF-α and α-SMA proteins(P<0.05).(4)After Cur intervention,the expression levels of mitophagy-related proteins increased and the expression levels of in TNF-α and α-SMA levels decreased in HepG2 cells induced by oleic acid and cholesterol(P<0.05).(5)Compared with M group,oleic-acidand cholesterol-induced mitophagy function in HepG2 cells was decreased after PINK1 knockdown(P<0.05).After CC inhibited AMPK,Cur increased the expression of p-AMPK(P<0.01),Sirt1(P<0.01),peroxisome proliferator-activated receptor γ coactivator-1α(P>0.05),PINK1(P<0.01)and parkin(P<0.01)proteins to some extent.CONCLUSION:Treatment with Cur attenuates liver injury in NASH mice and reduces lipid accumulation in HepG2 cells induced by oleic acid and cholesterol,and the mechanism may be related to promotion of mitophagy,which may involve the AMPK/Sirt1 signaling pathway.

Objective:To investigate the personnel, equipment, new technology of various radiotherapy units in Gansu Province, aiming to deeply understand the current status of radiotherapy in Gansu Province.Methods:From March 2023 to March 2024, the data of personnel, equipment and technology of different radiotherapy units in Gansu Province were investigated via online questionnaire survey and offline field visits by Gansu Provincial Quality Control Center for Radiation Oncology. The questionable data were reviewed, logical verification and outlier identification were conducted, and all information was entered according to the principle of double entry. Qualitative data were described by frequency (percentage).Results:As of March 2024, there are 26 radiotherapy units in Gansu Province, including 7 units in Lanzhou, capital city of Gansu Province and none in Linxia Hui Autonomous Prefecture and Gannan Tibetan Autonomous Prefecture. The ratio of radiotherapy physicians and physicists was 2.71 vs. 1, lower than the national level of 3.51 vs. 1. Radiotherapy physicians with intermediate and senior titles accounted for 66.2%, 54% for physicists with intermediate and senior titles and 34.0% for therapists with intermediate and senior titles, respectively. There were 1.70 accelerators per million population in Gansu Province. Among 42 accelerators in Gansu Province, 40 were linac and 2 heavy ion accelerators. In Lanzhou, 7 radiotherapy units had 17 linac, and the remaining 11 cities had 23 linac. All 40 linac could perform 3D conformal radiotherapy and intensity-modulated radiotherapy. The high-end equipment could apply advanced technologies such as volumetric modulated arc radiotherapy, stereotactic body radiotherapy, optical surface-guided radiation therapy and adaptive radiation therapy, etc. Relevant data in this survey were improved compared with those in the national surveys conducted in 2006, 2015 and 2019. Conclusions:The radiotherapy status has developed in Gansu Province, whereas uneven development still exists among regions. According to the actual demands of equipment, talents and technology in each region, the structural adjustment should be oriented to the underdeveloped cities and counties in the province to meet the local medical needs.

AIM:This study explores whether curcumin(Cur)promotes mitophagy to attenuate nonalcoholic steatohepatitis(NASH)in mice,as well as the possible molecular mechanisms involved.METHODS:A high-fat and high-cholesterol diet was used to replicate the NASH mouse model.Thirty-two male C57BL/6J mice were randomly divided into normal control(NC)group,high-fat and high-cholesterol model(M)group,M+low-dose Cur(Cur-L)group,and M+high-dose Cur(Cur-H)group,with 8 mice in each group.The weight of 8 mice in each group was recorded weekly.After feeding for 18 weeks,the serum and liver of mice were collected.Serum levels of total cholesterol(TC),triglyceride(TG),low-density lipoprotein(LDL-C),alanine aminotransferase(ALT),aspartate aminotransferase(AST),and tumor necrosis factors-α(TNF-α)were measured.Liver index was calculated,and steatosis,inflammation,and fibrosis of the liver were observed by HE and Masson staining.Western blot analysis was performed to detect the protein expression of mi-tophagy-related protein,TNF-α and α-SMA in the liver.(2)HepG2 cells were treated with oleic acid and cholesterol to replicate the hepatocyte injury model,which was divided into NC group,Cur group,M group,and M+Cur group.Small interfering RNA for PTEN-induced kinase 1(PINK1)knockdown was used to explore the relationship between PINK1-me-diated mitophagy and NASH.Compound C(CC)was used to inhibit AMP-activated protein kinase(AMPK)to explore the effect of the AMPK/silent information regulator 1(Sirt1)pathway on mitophagy.The lipid droplets of HepG2 cells were ob-served by oil red O staining,and the levels of TC,TG,LDL-C,ALT,and AST in cell suspension were detected.RE-SULTS:(1)Compared with M group,treatment with Cur significantly reduced the body weight,liver coefficient,and se-rum levels of TC,TG,LDL-C,ALT,AST,and TNF-α in NASH mice,while the steatosis and fibrosis in the liver were improved(P<0.05).(2)Different concentrations of Cur could increase or decrease the expression of mitophagy-related proteins in HepG2 cells in a concentration gradient.Compared with the M group,Cur reduced lipid droplets and de-creased TC,TG,LDL-C,ALT,and AST levels(P<0.05).(3)Compared with the NC group,the expression levels of mi-tophagy-related proteins in the liver of mice in the M group decreased,and the expression levels of TNF-α and α-SMA pro-teins increased.Different concentrations of Cur intervention promoted the increase of mitophagy-related proteins and the decrease of TNF-α and α-SMA proteins(P<0.05).(4)After Cur intervention,the expression levels of mitophagy-related proteins increased and the expression levels of in TNF-α and α-SMA levels decreased in HepG2 cells induced by oleic acid and cholesterol(P<0.05).(5)Compared with M group,oleic-acidand cholesterol-induced mitophagy function in HepG2 cells was decreased after PINK1 knockdown(P<0.05).After CC inhibited AMPK,Cur increased the expression of p-AMPK(P<0.01),Sirt1(P<0.01),peroxisome proliferator-activated receptor γ coactivator-1α(P>0.05),PINK1(P<0.01)and parkin(P<0.01)proteins to some extent.CONCLUSION:Treatment with Cur attenuates liver injury in NASH mice and reduces lipid accumulation in HepG2 cells induced by oleic acid and cholesterol,and the mechanism may be related to promotion of mitophagy,which may involve the AMPK/Sirt1 signaling pathway.

A 35-year-old female patient with advanced lung cancer was treated with paclitaxel(albumin-bound)and carboplatin chemotherapy,combined with tislelizumab immunotherapy for one cycle.After 6 days of treatment,the patient developed sagging of the right eyelid compared to the leftside,mild morning and heavy evening,systemic muscle soreness,abnormal transaminases and myocardial enzymes,and abnormal electrocardiogram.It is suspected that there may be a correlation between tislelizumab and myasthenia gravis or related myositis.After admission,patients were treated with methylprednisolone,intravenous immunoglobulin,plasma exchange,mycophenolate mofetil,temporary pacemaker installation,and tracheal intubation.After more than a month of treatment,the patient's indicators decreased,but the patient became unconscious,the patient's family requested discharge.The association between the multisystem adverse reactions and tislelizumab was evaluated using the Naranjo's Assessment Scale,the correlation score was 7 and evaluated as probably relevant.It is suggested that when using tislelizumab in clinical practice,risk factor assessment and medication monitoring should be strengthened to ensure drug safety.

A 35-year-old female patient with advanced lung cancer was treated with paclitaxel(albumin-bound)and carboplatin chemotherapy,combined with tislelizumab immunotherapy for one cycle.After 6 days of treatment,the patient developed sagging of the right eyelid compared to the leftside,mild morning and heavy evening,systemic muscle soreness,abnormal transaminases and myocardial enzymes,and abnormal electrocardiogram.It is suspected that there may be a correlation between tislelizumab and myasthenia gravis or related myositis.After admission,patients were treated with methylprednisolone,intravenous immunoglobulin,plasma exchange,mycophenolate mofetil,temporary pacemaker installation,and tracheal intubation.After more than a month of treatment,the patient's indicators decreased,but the patient became unconscious,the patient's family requested discharge.The association between the multisystem adverse reactions and tislelizumab was evaluated using the Naranjo's Assessment Scale,the correlation score was 7 and evaluated as probably relevant.It is suggested that when using tislelizumab in clinical practice,risk factor assessment and medication monitoring should be strengthened to ensure drug safety.

Objective:To investigate the personnel, equipment, new technology of various radiotherapy units in Gansu Province, aiming to deeply understand the current status of radiotherapy in Gansu Province.Methods:From March 2023 to March 2024, the data of personnel, equipment and technology of different radiotherapy units in Gansu Province were investigated via online questionnaire survey and offline field visits by Gansu Provincial Quality Control Center for Radiation Oncology. The questionable data were reviewed, logical verification and outlier identification were conducted, and all information was entered according to the principle of double entry. Qualitative data were described by frequency (percentage).Results:As of March 2024, there are 26 radiotherapy units in Gansu Province, including 7 units in Lanzhou, capital city of Gansu Province and none in Linxia Hui Autonomous Prefecture and Gannan Tibetan Autonomous Prefecture. The ratio of radiotherapy physicians and physicists was 2.71 vs. 1, lower than the national level of 3.51 vs. 1. Radiotherapy physicians with intermediate and senior titles accounted for 66.2%, 54% for physicists with intermediate and senior titles and 34.0% for therapists with intermediate and senior titles, respectively. There were 1.70 accelerators per million population in Gansu Province. Among 42 accelerators in Gansu Province, 40 were linac and 2 heavy ion accelerators. In Lanzhou, 7 radiotherapy units had 17 linac, and the remaining 11 cities had 23 linac. All 40 linac could perform 3D conformal radiotherapy and intensity-modulated radiotherapy. The high-end equipment could apply advanced technologies such as volumetric modulated arc radiotherapy, stereotactic body radiotherapy, optical surface-guided radiation therapy and adaptive radiation therapy, etc. Relevant data in this survey were improved compared with those in the national surveys conducted in 2006, 2015 and 2019. Conclusions:The radiotherapy status has developed in Gansu Province, whereas uneven development still exists among regions. According to the actual demands of equipment, talents and technology in each region, the structural adjustment should be oriented to the underdeveloped cities and counties in the province to meet the local medical needs.

AIM:The aim of this study was to investigate the effects and mechanism of high fat on autolyso-somes in hepatoma cells before and after serum deprivation.METHODS:HepG2 cells were intervened with 1 mmol/L so-dium oleate to create a cell high-fat model.The gene expression of transcription factor EB(TFEB)in HepG2 cells was knocked down using TFEB small interfering RNA(TFEB-siRNA)transfection reagent.AMP-activated protein kinase(AMPK)inhibitor compound C(CC)was used to inhibit AMPK phosphorylation expression in HepG2 cells.The expres-sion of nuclear and cytoplasmic TFEB,lysosome-associated membrane protein 1(LAMP1),microtubule-associated pro-tein light chain 3(LC3),autophagy adaptor protein(p62),AMPK,and p-AMPK proteins in each group was analyzed through Western blot experiments.Lipid metabolism and liver function damage in each group were analyzed using total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)assay kits.The accumulation of lipid droplets in each group of cells was analyzed through oil red O staining.RESULTS:(1)Sodium oleate intervention resulted in a concentration-dependent decrease in the protein expression levels of LAMP1,LC3-Ⅱ/LC3-Ⅰ,and nuclear TFEB,while increasing the protein expression level of p62(P＜0.01).(2)Compared to the NC group,the sodium oleate group showed decreased expression of LAMP1,LC3-Ⅱ/LC3-Ⅰ,nuclear TFEB,and AMPK phosphorylation levels,with an increase in p62 expression.Compared to the sodium oleate group,the sodium oleate+serum deprivation combined intervention group showed increased nuclear TFEB,LAMP1,LC3-Ⅱ/LC3-Ⅰ,AMPK phosphorylation levels,and decreased p62 expression levels(P＜0.05).(3)The levels of TC,TG,LDL-C,ALT,and AST were increased in the sodium oleate group compared to the NC group.Serum deprivation reduced the number of lipid droplets induced by sodium oleate in HepG2 cells and decreased the levels of TC,TG,LDL-C,ALT,and AST compared to the sodium oleate group(P＜0.05).(4)Knockdown of TFEB did not result in significant changes in the levels of nuclear TFEB,LAMP1,LC3-Ⅱ/LC3-Ⅰ,p62,TC,TG,LDL-C,ALT and AST compared to the so-dium oleate group.(5)Inhibition of AMPK phosphorylation did not result in significant changes in the levels of nuclear TFEB,LAMP1,LC3-Ⅱ/LC3-Ⅰ,p62,and AMPK phosphorylation in the sodium oleate+serum deprivation group com-pared to the sodium oleate group.CONCLUSION:Serum deprivation improves sodium oleate-induced lipid metabolism damage in HepG2 cells through the autophagolysosome pathway mediated by AMPK-TFEB.

AIM:The aim of this study was to investigate the effects and mechanism of high fat on autolyso-somes in hepatoma cells before and after serum deprivation.METHODS:HepG2 cells were intervened with 1 mmol/L so-dium oleate to create a cell high-fat model.The gene expression of transcription factor EB(TFEB)in HepG2 cells was knocked down using TFEB small interfering RNA(TFEB-siRNA)transfection reagent.AMP-activated protein kinase(AMPK)inhibitor compound C(CC)was used to inhibit AMPK phosphorylation expression in HepG2 cells.The expres-sion of nuclear and cytoplasmic TFEB,lysosome-associated membrane protein 1(LAMP1),microtubule-associated pro-tein light chain 3(LC3),autophagy adaptor protein(p62),AMPK,and p-AMPK proteins in each group was analyzed through Western blot experiments.Lipid metabolism and liver function damage in each group were analyzed using total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)assay kits.The accumulation of lipid droplets in each group of cells was analyzed through oil red O staining.RESULTS:(1)Sodium oleate intervention resulted in a concentration-dependent decrease in the protein expression levels of LAMP1,LC3-Ⅱ/LC3-Ⅰ,and nuclear TFEB,while increasing the protein expression level of p62(P＜0.01).(2)Compared to the NC group,the sodium oleate group showed decreased expression of LAMP1,LC3-Ⅱ/LC3-Ⅰ,nuclear TFEB,and AMPK phosphorylation levels,with an increase in p62 expression.Compared to the sodium oleate group,the sodium oleate+serum deprivation combined intervention group showed increased nuclear TFEB,LAMP1,LC3-Ⅱ/LC3-Ⅰ,AMPK phosphorylation levels,and decreased p62 expression levels(P＜0.05).(3)The levels of TC,TG,LDL-C,ALT,and AST were increased in the sodium oleate group compared to the NC group.Serum deprivation reduced the number of lipid droplets induced by sodium oleate in HepG2 cells and decreased the levels of TC,TG,LDL-C,ALT,and AST compared to the sodium oleate group(P＜0.05).(4)Knockdown of TFEB did not result in significant changes in the levels of nuclear TFEB,LAMP1,LC3-Ⅱ/LC3-Ⅰ,p62,TC,TG,LDL-C,ALT and AST compared to the so-dium oleate group.(5)Inhibition of AMPK phosphorylation did not result in significant changes in the levels of nuclear TFEB,LAMP1,LC3-Ⅱ/LC3-Ⅰ,p62,and AMPK phosphorylation in the sodium oleate+serum deprivation group com-pared to the sodium oleate group.CONCLUSION:Serum deprivation improves sodium oleate-induced lipid metabolism damage in HepG2 cells through the autophagolysosome pathway mediated by AMPK-TFEB.