Objective:To investigate the effect of uridine diphosphate ceramide galactosyltransferase 8(UGT8)on colorectal cancer(CRC)cell growth and migration,elucidate an underlying mechanism,and assess the potential regulatory role of SRY-box transcription factor 9(SOX9)on UGT8.Methods:UGT8 and SOX9 mRNA expression levels in CRC tissues,and correlation between their expression levels,were analyzed using GEPIA2,UALCAN,and TIMER 2.0 online databases.UGT8 and SOX9 protein expression in CRC and adjacent tissues was detec-ted using immunohistochemistry,and relationships between their expression and clinicopathological characteristics were analyzed.Impact of UGT8 knockdown on CRC cell proliferation was assessed using a CCK-8 assay,and cell migration was evaluated using Transwell and wound healing assays.Western blot was performed to detect expression of epithelial-mesenchymal transition(EMT)markers(E-cadherin and ZEB1).RT-qPCR and Western blot were used to measure UGT8 mRNA and protein expression levels after SOX9 knockdown.The JASPAR online database was used to assess SOX9 potential for binding to the UGT8 promoter.Results:Bioinformatics analyses revealed significantly higher mRNA expression levels of both UGT8 and SOX9 in CRC tissues than in normal tissues.Positive correlation was observed between expres-sion levels.Immunohistochemistry results showed that tumor UGT8 and SOX9 protein levels were significantly higher than those in adjacent tissues.UGT8 protein level was found to correlates with N stage,and SOX9 protein level correlated with T stage.A positive correlation was observed between UGT8 and SOX9 expression levels.Following UGT8 knockdown,cell proliferation capacity was attenuated and cell migra-tion ability was reduced.E-cadherin expression concurrently increased and ZEB1 expression decreased.RT-qPCR and Western blot results showed that SOX9 knockdown significantly reduced UGT8 mRNA and protein levels.The JASPER website predicts that SOX9 will bind to the UGT8 promoter.Conclusions:UGT8 and SOX9 are highly expressed in CRC tissues,and their expression levels correlate with clinicopatholo-gical features.UGT8 and SOX9 expression levels display significant positive correlation.Mechanistically,UGT8 promotes CRC cell prolifera-tion and migration by facilitating epithelial-mesenchymal transition(EMT).SOX9 enhances UGT8 mRNA and protein expression and may bind to the UGT8 promoter region.

Objective:To investigate the effect of uridine diphosphate ceramide galactosyltransferase 8(UGT8)on colorectal cancer(CRC)cell growth and migration,elucidate an underlying mechanism,and assess the potential regulatory role of SRY-box transcription factor 9(SOX9)on UGT8.Methods:UGT8 and SOX9 mRNA expression levels in CRC tissues,and correlation between their expression levels,were analyzed using GEPIA2,UALCAN,and TIMER 2.0 online databases.UGT8 and SOX9 protein expression in CRC and adjacent tissues was detec-ted using immunohistochemistry,and relationships between their expression and clinicopathological characteristics were analyzed.Impact of UGT8 knockdown on CRC cell proliferation was assessed using a CCK-8 assay,and cell migration was evaluated using Transwell and wound healing assays.Western blot was performed to detect expression of epithelial-mesenchymal transition(EMT)markers(E-cadherin and ZEB1).RT-qPCR and Western blot were used to measure UGT8 mRNA and protein expression levels after SOX9 knockdown.The JASPAR online database was used to assess SOX9 potential for binding to the UGT8 promoter.Results:Bioinformatics analyses revealed significantly higher mRNA expression levels of both UGT8 and SOX9 in CRC tissues than in normal tissues.Positive correlation was observed between expres-sion levels.Immunohistochemistry results showed that tumor UGT8 and SOX9 protein levels were significantly higher than those in adjacent tissues.UGT8 protein level was found to correlates with N stage,and SOX9 protein level correlated with T stage.A positive correlation was observed between UGT8 and SOX9 expression levels.Following UGT8 knockdown,cell proliferation capacity was attenuated and cell migra-tion ability was reduced.E-cadherin expression concurrently increased and ZEB1 expression decreased.RT-qPCR and Western blot results showed that SOX9 knockdown significantly reduced UGT8 mRNA and protein levels.The JASPER website predicts that SOX9 will bind to the UGT8 promoter.Conclusions:UGT8 and SOX9 are highly expressed in CRC tissues,and their expression levels correlate with clinicopatholo-gical features.UGT8 and SOX9 expression levels display significant positive correlation.Mechanistically,UGT8 promotes CRC cell prolifera-tion and migration by facilitating epithelial-mesenchymal transition(EMT).SOX9 enhances UGT8 mRNA and protein expression and may bind to the UGT8 promoter region.

Objective:To investigate the effectiveness of new cryoprotective agents in preserving and transplanting human adipose tissue.Methods:The adipose tissue samples were obtained from healthy adult females who underwent liposuction at the Department of Plastic Surgery of Peking University Third Hospital from January to March 2022. The adipose tissue samples were centrifuged and then randomly divided into 9 groups. These groups were cryopreserved in liquid nitrogen using different cryoprotective agents [group A, group B, and dimethyl sulfoxide (DMSO) group] and cryopreservation times (1-month, 2-month, and 3-month groups), respectively. The cryoprotective agent formulation in group A was dextrose glycoside 40 (DEX), amino acids, vitamins, and inorganic salts. In group B, the formulation included DMSO and DEX. The ratio of cryoprotective agent in the DMSO group was 10% DMSO, 20% fetal bovine serum (FBS), and 70% DMEM-12. For cryopreservation, 5 ml cryogenic tubes were used with a fat to cryoprotective agent ratio of 3∶2, and each group contains 6 tubes for cryopreservation. After thawing the adipose tissue, HE staining was used to observe the histological morphology. Immunohistochemical staining was employed for the quantitative analysis of lipid droplet-encapsulated protein (Perilipin), and the Perilipin positivity rate was calculated by the ratio of the number of positive cells to the total number of cells. Adipocyte viability was assessed using the CCK-8 method. Thirty-eight healthy, clean nude mice were selected and divided into 3 groups of 12 mice each according to the use of different cryoprotective agents (groups A, B, and DMSO), while the other 2 mice were used as the day 0 control group. The mean fat freezing duration for all groups was 3 months. After nude mice were anesthetized intraperitoneally, 0.9 ml of thawed cryopreserved fat was injected into the dorsum bilaterally. The rate of adipose tissue retention was calculated by MRI scanning and three-dimensional software at 1, 2, and 3 months after transplantation, and compared between the groups. The fat grafts were explanted from the mice after they were sacrificed, and then subjected to histological morphology and quantitative analysis of Perilipin by using HE staining and immunohistochemical staining. GraphPad Prism 8.0 software was used for statistical analysis of the data. The data that conformed to a normal distribution were expressed as Mean ± SD. The overall comparison between multiple groups used analysis of variance for repeated measures. The comparison of data between groups at the same time point used Tukey’s multiple comparison test.Results:The morphology of adipose tissue in different cryoprotective agent groups closely resembled that of normal fresh adipose tissue after being cryopreserved in liquid nitrogen for 1-3 months. The difference in the proportion of Perilipin-stained positive cells in each group was not statistically significant ( P>0.05). The CCK-8 method indicated that the effect of the DMSO group was superior to groups A and B at 1 and 3 months of cryopreservation ( P<0.01), and that the DMSO group and group B were superior to group A at 2 months of cryopreservation ( P<0.01). In the animal experiments, there was no statistically significant difference between the groups in the volume retention rate 1-3 months after cryopreserved fat transplantation ( P>0.05). Additionally, the adipose tissues in each group exhibited varying degrees of localized necrosis accompanied by an inflammatory reaction 1-3 months after transplantation. There was no statistically significant difference in the Perilipin staining positivity between the groups ( P>0.05). Conclusion:The use of new cryoprotective agents for cryopreserving adipose tissue does not show a significant difference compared to the traditional cryoprotective agent. However, it is theoretically safer as it avoids the potential toxic effects of using DMSO or FBS on the human body.

Objective:To investigate the effectiveness of new cryoprotective agents in preserving and transplanting human adipose tissue.Methods:The adipose tissue samples were obtained from healthy adult females who underwent liposuction at the Department of Plastic Surgery of Peking University Third Hospital from January to March 2022. The adipose tissue samples were centrifuged and then randomly divided into 9 groups. These groups were cryopreserved in liquid nitrogen using different cryoprotective agents [group A, group B, and dimethyl sulfoxide (DMSO) group] and cryopreservation times (1-month, 2-month, and 3-month groups), respectively. The cryoprotective agent formulation in group A was dextrose glycoside 40 (DEX), amino acids, vitamins, and inorganic salts. In group B, the formulation included DMSO and DEX. The ratio of cryoprotective agent in the DMSO group was 10% DMSO, 20% fetal bovine serum (FBS), and 70% DMEM-12. For cryopreservation, 5 ml cryogenic tubes were used with a fat to cryoprotective agent ratio of 3∶2, and each group contains 6 tubes for cryopreservation. After thawing the adipose tissue, HE staining was used to observe the histological morphology. Immunohistochemical staining was employed for the quantitative analysis of lipid droplet-encapsulated protein (Perilipin), and the Perilipin positivity rate was calculated by the ratio of the number of positive cells to the total number of cells. Adipocyte viability was assessed using the CCK-8 method. Thirty-eight healthy, clean nude mice were selected and divided into 3 groups of 12 mice each according to the use of different cryoprotective agents (groups A, B, and DMSO), while the other 2 mice were used as the day 0 control group. The mean fat freezing duration for all groups was 3 months. After nude mice were anesthetized intraperitoneally, 0.9 ml of thawed cryopreserved fat was injected into the dorsum bilaterally. The rate of adipose tissue retention was calculated by MRI scanning and three-dimensional software at 1, 2, and 3 months after transplantation, and compared between the groups. The fat grafts were explanted from the mice after they were sacrificed, and then subjected to histological morphology and quantitative analysis of Perilipin by using HE staining and immunohistochemical staining. GraphPad Prism 8.0 software was used for statistical analysis of the data. The data that conformed to a normal distribution were expressed as Mean ± SD. The overall comparison between multiple groups used analysis of variance for repeated measures. The comparison of data between groups at the same time point used Tukey’s multiple comparison test.Results:The morphology of adipose tissue in different cryoprotective agent groups closely resembled that of normal fresh adipose tissue after being cryopreserved in liquid nitrogen for 1-3 months. The difference in the proportion of Perilipin-stained positive cells in each group was not statistically significant ( P>0.05). The CCK-8 method indicated that the effect of the DMSO group was superior to groups A and B at 1 and 3 months of cryopreservation ( P<0.01), and that the DMSO group and group B were superior to group A at 2 months of cryopreservation ( P<0.01). In the animal experiments, there was no statistically significant difference between the groups in the volume retention rate 1-3 months after cryopreserved fat transplantation ( P>0.05). Additionally, the adipose tissues in each group exhibited varying degrees of localized necrosis accompanied by an inflammatory reaction 1-3 months after transplantation. There was no statistically significant difference in the Perilipin staining positivity between the groups ( P>0.05). Conclusion:The use of new cryoprotective agents for cryopreserving adipose tissue does not show a significant difference compared to the traditional cryoprotective agent. However, it is theoretically safer as it avoids the potential toxic effects of using DMSO or FBS on the human body.

OBJECTIVE: To predict the incidence of lung cancer in Jiashan county from 2017 to 2019 on the basis of the incidence rates of lung cancer during 1987-2016. METHODS: Lung cancer incident cases were derived from cancer registry system of Jiashan. Crude incidence, age-standardized incidence rate by the Chinese standard population (ASR China) and the world standard population (ASR world) were calculated. Annual percent change (APC) was used to examine the temporal trend, and the autoregressive integrated moving average method (ARIMA) of time series model was used to predict the incidence rates from 2017 to 2019. RESULTS: There were 6103 lung cancer incident cases during 1987-2016 in Jiashan county. Averagely, the crude incidence rate, ASR China and ASR world were 53.77/10, 25.24/10 and 34.15/10, respectively. The crude incidence rate, ASR China and ASR world in male were 78.30/10, 34.77/10 and 51.87/10, which were higher than those in female (29.15/10, 14.31/10 and 17.99/10). Crude incidence rate increased from 27.58/10 in 1987 to 111.24/10 in 2016, and the APC was 5.28%. Crude incidence rate predicted by ARIMA model from 2017 to 2019 would be 135.64/10, 145.97/10 and 152.63/10, and the predicted crude incidence rate for 2017 was close to the real incidence rate in 2017 (135.95/10). CONCLUSIONS The incidence of lung cancer in Jiashan has been increased dramatically over the past 30 years and will continue to increase in the future.
China ; epidemiology ; Female ; Humans ; Incidence ; Lung Neoplasms ; epidemiology ; Male ; Registries

OBJECTIVE To explore the effect of Lianhuaqingwen capsules ( LHQW ) on junction protein expression in mouse lung tissue of lipopolysaccharide (LPS)-induced acute lung injury ( ALl). METHODS 120 male mice were randomly divided into six groups: normal control, model, model+dexa-methasone 5 mg.kg-1 , model +LHQW 2, 4 and 8 g.kg-1 groups. Dexamethasone and LHQW were administered orally, once daily, for 7 d. 24 h after the last administration, LPS solution was instilled into the tracheas of mice except the normal control group to prepare the mouse model of ALl. 24 h after the establishment of the ALl model, the mice were sacrificed and the pathological changes in the mouse lung tissue were observed by optical microscopy and ultrastructure of alveolar epithelium was observed by transmission electron microscopy. The cell percentage of positive expression of tumor necrosis factor-α(TNF-α) in the peripheral blood T lymphocytes was detected by flow cytometry. The expressions of con-nexin 43 ( Cx43), occludin and zonula occludens protein-1 ( ZO-1) in lung tissues were detected by immunohistochemistry. RESULTS Under the light microscope, the mouse lung of model group showed a large amount of inflammatory cell infiltration and alveolar wall thickening. Compared with model group, inflammatory cell infiltration was reduced in model+dexamethasone, model+LHQW 2,4 and 8 mg.kg-1 groups. Under the electron microscope, the mouse alveolar epithelial cells of model group showed injury. Compared with model group, the damage was reduced in model+dexamethasone, and model+LHQW 2, 4 and 8 mg.kg-1 groups. The cell percentage of TNF-α positive expression in peripheral blood T lympho-cytes in normal control, model, model+dexamethasone, model+LHQW 2,4 and 8 mg.kg-1 groups was (3.6±0.9)%, (6.4±0.8)%, (2.8±0.7)%, (4.7±1.6)%, (4.0±1.5)% and (3.6±1.2)%, respectively. The percentage in model group was obviously higher than that in normal control group( P<0.01), but was lower in the four drug treatment groups than in model group(P<0.05, P<0.01). The expression of Cx43, occludin and ZO-1 in lung tissue of model group was lower than that of normal control group(P<0.01), but higher in model+dexamethasone, model + LHQW 4 and 8 mg.kg-1 groups than in model group(P<0.05). CONCLUSION LHQW may alleviate ALl induced by LPS and play a protective role by inhibiting inflammatory cell infiltration and improving protein connection expression in alveolar epithelial cells and pulmonary vascular endothelial cells.

AIM:To investigate the role of anti-miR-21 oligonucleotide ( AMO) in the anti-leukemic activity of decitabine (DCA) in vitro.METHODS:AMO and scramble oligonucleotide (SCR) were constructed and transfected into HL-60 cells.The miR-21 expression was analyzed by real-time PCR to identify the transfection efficiency .The cells were treated with DCA at gradient concentrations (0.5, 2.0 and 4.0 μmol/L) for 48 h.The mRNA expression of human period circadian protein 3 (hPer3) was detected by real-time PCR.The early apoptotic rates were determined by flow cy-tometry with Annexin V/PI staining.Mean fluorescence intensities ( MFI) of CD117 and CD11b were also measured by flow cytometry.RESULTS:The miR-21 relative expression level in AMO group was significantly lower than that in blank group and SCR group (P<0.01).IC50 of DCA in AMO group was significantly lower than that in blank group and SCR group (P<0.01).With the same concentration of DCA, the early apoptotic rate, the mRNA expression of hPer3 and the MFI of CD11b in AMO group were significantly higher than those in blank group and SCR group (P<0.01).The MFI of CD117 in AMO group were significantly lower than those in blank group and SCR group ( P<0.01 ) .CONCLUSION:Activation of hPer3 expression plays an important role in enhanced anti-leukemic activity of decitabine by AMO in vitro.

Aim To investigate the effect of Jinlida on cholesterol-related genes in skeletal muscle in fat-in-duced insulin resistance ApoE-/ - mice. Methods Ten male C57 BL/6 J mice were selected as normal group ( NF );50 male ApoE-/ - mice with a high-fat feeding after 16 weeks ( HF) were divided into model group, rosiglitazone ( LGLT ) , Jinlida low dose group ( JLDL, 0. 95 g · kg-1 · d-1 ) , Jinlida medium dose group ( JLDM, 1. 9 g·kg-1 ·d-1 ) , Jinlida high dose group (JLDH, 3. 8 g·kg-1·d-1), which were per-formed intragastric administration for 8 weeks. Oil red O staining of mouse skeletal muscle was used for fat ac-cumulation. Insulin receptor ( INSR) , insulin receptor body substrate-1 ( IRS-1 ) , low-density lipoprotein re-ceptor ( LDLR ) , cholesterol sensor ( SCAP ) mRNA and protein expression in mouse skeletal muscle were measured by quantitative reverse transcription PCR ( RT-PCR ) and Western blot. Results Compared with NF group, fasting blood glucose ( FBG) , choles-terol ( TC ) , triglyceride ( TG ) and low density lipo-protein cholesterol ( LDL-C ) of HF mice were signifi-cantly elevated, while high-density lipoprotein ( HDL-C ) significantly decreased ( P < 0. 05 ) . Compared with HF group, Jinlida group could reduce to varying degrees FBG, TC, TG and LDL-C in mice, and in-crease HDL-C ( P <0. 05 ) . Jinlida could downgrade fasting serum insulin ( FINS ) level, and improve the insulin sensitive index ( ISI ) ( P < 0. 05 ) . Jinlida could obviously improve skeletal muscle fat accumula-tion of mice. Compared with NF group, skeletal mus-cle INSR, IRS-1, LDLR mRNA and protein levels of HF group were significantly decreased ( P <0. 05 ) , while SCAP mRNA and protein level increased signifi-cantly (P<0. 05). Compared with HF group, Jinlida could increase to varying degrees INSR, IRS-1, LDLR mRNA and protein levels ( P < 0. 05 ) , and lower SCAP mRNA and protein levels ( P<0. 05 ) . Conclu-sion Jinlida can alleviate fat-induced insulin resist-ance in ApoE-/ - mice through regulation of cholester-ol-related gene expression.

OBJECTIVETo evaluate the sensitivity and specificity of optimized sequential screening program of colorectal cancer, and provide evidence for the further optimization of colorectal cancer screening program.

METHODSUsing cluster sampling method, 4 administrative villages were selected from Jiashan county as a census district in 2011 to 2013. Volunteers of 40 to 74 years old in the census were recruited, and tested by both optimized sequential screening (including questionnaire survey and fecal occult blood test) and colonoscopy for colorectal cancer. Sensitivity and specificity of different screening methods were calculated, respectively.

RESULTSA total of 2 607 volunteers took both simultaneously screening and colonoscopy at the same time. 20 colorectal cancer cases, 85 advanced adenoma cases, 271 non-advanced adenomas cases, and 141 non-adenomatous polyps cases were detected. Sensitivity of optimized sequential screening for colorectal cancer, advanced adenomas, and non-advanced adenomas were 70.0% (14/20) , 57.6% (49/85) and 36.5% (99/271) , specificity was 68.7% (1 776/2 587) , 69.2% (1 746/2 522) and 68.9% (1 610/2 336) , respectively. Sensitivity of the fecal occult blood test of colorectal cancer, advanced adenomas and non-advanced adenomas were 70.0% (14/20) , 47.1% (40/85) and 26.6% (72/271), specificity was 79.4% (2 053/2 587), 79.9% (2 014/2 522) and 79.6% (1 860/2 336). The sensitivity of fecal occult blood test and those of optimized sequential screening for colorectal cancer, advanced adenomas was not significant (χ(2) = 0.00, 1.91, all P values > 0.05). Sensitivity of questionnaire survey of colorectal cancer, advanced adenomas and non-advanced adenomas were 10.0% (2/20), 14.1% (12/85), 12.9% (35/271), specificity was 87.6% (2 266/2 587), 87.7% (2 211/2 522), 87.6% (2 046/2 336). There were no significant difference between non-advanced adenomas. The sensitivity of advanced adenomas and non-advanced adenomas showed no significant decline when the following six term were removed from screening programs: chronic diarrhea, chronic constipation, mucus or bloody history, history of chronic appendicitis or appendectomy surgery, chronic cholecystitis or gallbladder surgery, adverse events in the history of life, while the sensitivity of colorectal cancer remained nearly the same 70.0% (14/20), 52.9% (45/85), 31.4% (85/271) (χ(2) = 0.38, 1.61, all P values > 0.05).

CONCLUSIONCurrent optimized sequential screening programs for colorectal cancer in China have a high sensitivity and specificity. However, further optimization is viable and necessary.

Adenoma ; China ; Colonic Polyps ; Colonoscopy ; Colorectal Neoplasms ; Early Detection of Cancer ; Humans ; Mass Screening ; Occult Blood ; Sensitivity and Specificity ; Surveys and Questionnaires

Objective To investigate the FH/Wjd rat model of alcoholic liver disease.Methods Thirty-six 16-18 week old SPF grade FH/Wjd rats (male:female=1:1) were used in this study.The rats were divided into two groups randomly by body weight:water intake group and alcohol intake group.The rats took water or alcohol freely.16 weeks lat-er, ALT, AST, TBIL, TG, CHO in the serum and TG, GSH in the liver homogenate were detected.The expression of PPARαprotein in the liver tissue was detected by Western blot.The apoptosis rate of liver cells was assessed by flow cy-tometry.The pathological changes of liver tissue were examined using HE staining.Results Compared with the water in-take group, the serum TBIL and TG were significantly increased in rats of both sexes of the alcohol intake group, moreover, ALT and CHO of the female rats in the alcohol intake group were significantly decreased.TG in the liver homogenate in-creased obviously, while GSH in the liver homogenate showed a decreasing tendency.Hepatocyte apoptosis in rats of both sexes in the alcohol intake group showed an increasing tendency.The PPARαprotein expression was up-regulated obvious-ly, and the main pathological change in the liver tissue was microvesicular fatty degeneration.Conclusion Spontaneous long-term alcohol intake can induce liver injuries in FH/Wjd rats.