Ketosis in dairy cows is often accompanied by apoptotic and inflammatory responses in adipose tissue.In order to investigate the effect of pro-apoptotic protein Bid on adipocyte apoptosis in cows with ketosis,the adipose tissue was stained by TUNEL staining technique in this study to observe the apoptotic changes in adipose tissue of cows with ketosis.In the in vivo test,protein ex-pression of apoptosis-related factors Bid,Bax,C-Caspase-3,Bcl-2 and inflammation marker factors C1q,IL-1β,IL-10 and IL-6 in adipose tissues of healthy cows and ketosis cows were detected by Western blot.In the in vitro assay,the adipocyte lipolysis model was constructed by culturing pri-mary bovine adipocytes in vitro to inhibit Bid and adding isoproterenol(ISO),and the protein ex-pression levels of apoptosis-related molecules and inflammation-related molecules in adipocytes were detected by Western blot technique.The results of TUNEL staining showed that the protein expression of pro-apoptotic factors Bid,Bax and C-Caspase-3,and pro-inflammatory markers TNF-a,IL-1β,and IL-6 were significantly higher,and the protein expression of complement C1q,anti-ap-optotic factor Bcl-2,and anti-inflammatory factor IL-10 were significantly lower in adipose tissues of ketosis cows compared with that of healthy cows.The in vitro results showed that the protein expression levels of apoptosis and inflammation-related factors in adipocytes treated with the ISO group were significantly higher compared with those in the control group,while the protein ex-pression levels of apoptosis and inflammation-related factors in adipocytes treated with the addi-tion of the Bid inhibitor group were significantly lower.The above results showed that inhibition of Bid could alleviate the apoptotic and inflammatory responses of ISO on adipocytes.This will fur-ther clarify the role of Bid/C1q in the regulation of adipose tissue and cell apoptosis and inflamma-tion in ketosis cows.

OBJECTIVE: To explore the genetic basis for four patients suspected for Marfan syndrome (MFS). METHODS: Four male patients with suspected MFS and their family members who were treated at West China Second Hospital of Sichuan University from September 12, 2019 to March 27, 2021 were selected as the study subjects. Peripheral venous blood samples were collected from the patients and their parents or other pedigree members for the extraction of genomic DNA. Whole exome sequencing was carried out, and candidate variants were validated by Sanger sequencing. The pathogenicity of the variants was determined based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). RESULTS: Genetic testing revealed that all four patients have harbored variants of the FBN1 gene, including c.430_433del (p.His144fs) deletional variant in exon 5, c.493C>T (p.Arg165*) nonsense variant in exon 6, c.5304_5306del (p.Asp1768del) deletional variant in exon 44 and c.5165C>G (p.Ser1722Cys) missense variant in exon 42. According to the ACMG guidelines, the c.430_433del and c.493C>T were classified as pathogenic variants (PVS1+PM2_Supporting+PP4; PVS1+PS1+PS2+PM2_Supporting+PP4). c.5304_5306del and c.5165C>G were classified as likely pathogenic variants (PS2+PM2_Supporting+PM4+PP4; PS2_Moderate+PS1+PM1+PM2_Supporting). CONCLUSION The c.430_433del and c.5304_5306del variants of the FBN1 gene identified in this study were unreported previously. Above results have enriched the variation spectrum of the FBN1 gene and provided a basis for genetic counseling and prenatal diagnosis of patients with MFS and acromicric dysplasia.
Female ; Pregnancy ; Humans ; Male ; Exons ; China ; Family ; Genetic Counseling ; Genetic Testing ; Marfan Syndrome/genetics* ; Mutation ; Fibrillin-1/genetics*

AIM: To investigate the effects of phorbol ester (TPA) on the anti-tumor effect and renal toxicity of cisplatin (CP). METHODS: MTT assay was used to examine the effect of TPA on the proliferation inhibition of CP in A549 and SPC-A-1 lung cancer cells. Also the effect of TPA on acute toxicity of CP was observed by once injection of high dose CP through caudal vein; The tumor-bearing mice model was explored to investigate the effect of TPA on tumor inhibition ratio and renal toxicity of CP in vivo. And the effect of TPA on renal oxidative stress induced by CP was detected. RESULTS: 1 ng/mL TPA could significantly enhance the inhibitory effect of CP on cell proliferation. In acute toxicity test, TPA could significantly reduce the toxicity of CP and prolong the survival time of animals. And the tumor weight (P<0.05), serum creatinine (P<0.05) and urea nitrogen levels (P<0.01) in TPA combined with CP group were significantly lower than those in CP group. Meanwhile, the results of HE staining showed that the renal tissue damage was significantly reduced in the combined group compared with CP group. The contents of MDA in renal tissue were decreased (P<0.01). However, the contents of GSH and the activity of SOD were increased (P<0.05) in TPA and CP combined group. CONCLUSION: TPA can enhance the inhibitory effect of CP on cell proliferation and inhibit tumor growth in tumor-bearing mice. At the same time, TPA can reduce the renal toxicity of CP, which may be related to the inhibition of renal oxidative stress induced by CP.

Objective:To investigate the effects of PDCA method on improving the accuracy of the Autar Deep Venous Thrombosis (DVT) Scale used by nurses to assess the risk of deep venous thrombosis in surgical patients.Methods:Applied the way of PDCA, namely, raising questions, analyzing reasons, implementing measures, feeding back effects to professional explanate the difficulty in using the scale, moreover, training the nurses used level education method between August 2017 and December2018.The accuracy and consistency of the scale were compared before and after interventions.Results:A total 396 patients were evaluated in the study. There were significant differences in the consistency of evaluation among hospitalized, post-operative and discharged patients after intervention compared with before intervention ( P< 0.0167).The consistency of assessment of team members and responsible nurses increased from 68 to 120 cases, and the consistency of assessment reached 90.91%. The Kappa consistency of assessment of responsible nurses and team members increased from 0.354 before intervention to 0.879 after intervention. The effect was remarkable. Conclusions:PDCA cycle method is based on the problems found in clinical practice. It clarifies the professional terms and rules used in the scale, improves the accuracy and consistency of the Autar DVT scale used by nurses, correctly reflects the risk of thrombosis, ensures the safety of patients, and is worthy of promotion and reference.

Objective:To evaluate the role of CXC chemokine receptor 6 (CXCR6)-mediated activation of natural killer T (NKT) cells in renal fibrosis following acute kidney injury (AKI) in mice.Methods:Eighteen male wild-type C57BL/6 mice and 18 CXCR6 knockout C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 6 groups ( n=6 each) using a random number table method: wild-type mouse control group (group WT-CON), CXCR6 knockout mouse control group (group CXCR6 -/--CON), wild-type mouse with AKI group (group WT-AKI), CXCR6 knockout mouse with AKI group (group CXCR6 -/--AKI), wild-type mouse with AKI + NKT cell adoptive transfer group (group WT-AKI-NKT) and CXCR6 knockout mouse with AKI + NKT cell adoptive transfer group (group CXCR6 -/--AKI-NKT). Folic acid 250 mg/kg was intraperitoneally injected to establish the model of renal fibrosis in mice with AKI.NKT cellsuspension 250 μl(1×10 6 cells) was injected through the tail vein on the 4th and 9th days after folic acid injection in group WT-AKI-NKT and group CXCR6 -/--AKI-NKT, respectively.Blood samples were taken from orbital at day 14 after folic acid injection for determination of the concentrations of serum blood urea nitrogen (BUN) and creatinine (Cr). The animals were sacrificed, and renal tissues were obtained for observation of the area of renal fibrosis (by Sirius red staining) and renal injury (using H&E staining) which was scored and for determination of the proportion of CD1d Tetramer+ cells (by flow cytometry), the number of CD206 and α-smooth muscle actin (α-SMA) double positive (CD206 + -α-SMA + ) cells (by immunofluorescence) and expression of interleukin (IL)-4 and IL-13 mRNA (by real-time polymerase chain reaction). Results:Compared with group WT-CON, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group WT-AKI and WT-AKI-NKT ( P<0.05). Compared with group WT-AKI, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group WT-AKI-NKT ( P<0.05), and the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly decreased, and the expression of IL-4 and IL-13 mRNA was down-regulated in group CXCR6 -/--AKI ( P<0.05). Compared with group CXCR6 -/--CON, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased in group CXCR6 -/--AKI and group CXCR6 -/--AKI-NKT ( P<0.05). Compared with group CXCR6 -/--AKI, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group CXCR6 -/--AKI-NKT ( P<0.05). Conclusion:CXCR6-mediated activation of NKT cells is involved in renal fibrosis following AKI in mice, and the mechanism may be related to promoting Th2 cytokine-mediated M2 macrophage-myofibroblast transformation.

Objective: To evaluate the effect of dexmedetomidine on hypoxia-inducible factor-1alpha (HIF-1 α) signaling pathway during hypoxia in mice with lung cancer. Methods: Eighteen clean-grade healthy adult male BALB/c nude mice, aged 8 weeks, weighing 20-30 g, were divided into 3 groups (n=6 each) using a random number table method: lung cancer group (group L), hypoxia+ lung cancer group (group HL), and hypoxia plus lung cancer plus dexmedetomidine group (group HLD). Human lung adenocarcinoma A549 cell suspension 1×10⁶/150 μl was injected via the tail vein to establish the mouse model of lung cancer.After the model was established successfully, chronic intermittent hypoxia was performed as follows: the mice were placed in air-tight modular incubation chambers, and the atmosphere was controlled by a constant gas flow containing 10% O₂ for 3 h once a day for 3 weeks.The mice in group HL were exposed to hypoxia.After the end of hypoxia exposure, the animals in group HLD were intraperitoneally injected with dexmedetomidine 25 μg/kg 3 times a week for 3 weeks in total.The mice in group L were placed in air-tight modular incubation chambers and the atmosphere was controlled by a constant gas flow containing 21% O₂ for 3 h once a day for 3 weeks, and the equal volume of normal saline was injected intraperitoneally.The mice were sacrificed by cervical dislocation, and the lung tissues were removed for determination of lung cancer nodules count, HIF-1α expression (by immunohistochemistry), expression of HIF-1α, survivin and X chromosome linked inhibitor of apoptosis protein (XIAP) (by Western blot), and expression of matrix metalloproteinase-2 (MMP-2) and MMP-9 (by real-time polymerase chain reaction). Results: Compared with group L, the number of lung cancer nodules was significantly increased, and the expression of HIF-1α, survivin, XIAP, MMP-2 and MMP-9 was up-regulated in the other two groups (P<0.05). Compared with group HL, the number of lung cancer nodules was significantly increased, and the expression of HIF-1α, survivin, XIAP, MMP-2 and MMP-9 was up-regulated in group HLD (P<0.05). A small number of HIF-1α positive cells were found in group L, a medium number of HIF-1α positive cells in group HL, and a large number of HIF-1α positive cells in group HAD. Conclusion Dexmedetomidine can promote proliferation of tumor cells in mice with lung cancer during hypoxia, and the mechanism is related to activating HIF-1α signaling pathway.

Objective To investigate the effect and mechanism of suberoylanilide hydroxamic acid (SAHA) on the fear extinction in mice with chronic social defeat stress (SD). Methods Fifty-six male C57BL/6J mice aged 7-8 weeks were randomly divided into control group,social defeat group,control-SAHA group and social defeat-SAHA group to investigate the effect of SAHA and social defeat group,social defeat-AAV BDNF group and social defeat-AAV blank group to investigate the effect of BDNF. Fear extinction in mice was evaluated by fear conditioning test (FC). The levels of BDNF and HDAC2 in mice hippocampus were detected by Western blot (WB). The expression of BDNF-overexpressing virus in hippocampus of mice was detected by immunofluorescence assay. Results (1) Compared with control group,fear extinction in the social defeat group was significantly decreased (P<0. 05). Compared with control group, the level of HDAC2(0. 50±0. 02) in the social defeat group was significantly increased (P<0. 001),while the level of BDNF(0. 16 ± 0. 03) was significantly decreased (P<0. 001) in the social defeat group. ( 2) After using SAHA,fear extinction of mice significantly improved (P<0. 05). Compared with control group,the level of HDAC2 (0. 26±0. 02) in the control-SAHA group was significantly decreased(P<0. 001),and the level of BDNF (0. 40±0. 03) was significantly increased (P<0. 001). Compared with social defeat group,the level of HDAC2 (0. 39±0. 03) in the social defeat-SAHA group was significantly decreased (P<0. 001),and the lev-el of BDNF (0. 28±0. 01) was significantly increased (P<0. 001). (3)After injection BDNF-overexpressing virus,fear extinction was significantly improved(P<0. 05). Conclusion SAHA can enhance fear extinction in mice with chronic social defeat stress and its mechanism may be related to the up-regulation of BDNF ex-pression in hippocampus by inhibiting HDAC2 in hippocampal.

Objectives To investigate the effects of cell aging on the disorders relating to gastric mucosa aging.Methods A treatment of 200 μmol/L H2O2 was used to induce senescence of human gastric epithelial cell line GES-1,and the cell growth curve was monitored.Senescence secretory phenotypes were observed by detecting the protein level of p53 and p16INK4a with senescence-associated β-galactosidase(SA-β gal)staining and Western blot testing.The mRNA levels of senescence-associated secretory phenotype(SASP)factors in human gastric epithelial GES-1 cell including IL-1β,IL-6,IL-8,TGF-β、IFN-γ,and VEGF-A were detected by RT PCR.The mRNA expression levels of IL-1β,IL-6,IL-8,TGF-β,IFN-γ,and VEGF in the conditioned medium were detected by ELISA analysis.Results The 200 μmol/L H2O2-induced GES-1 cells stopped proliferating after 3 days of treatment,and cells enlarged and flattened at 10 days.The increased SA-β-gal staining(P＜0.001) and the increased expression levels of p53 and p16INK4a proteins indicated the success of establishing the aging model of GES-1.The mRNA levels of IL 1β,IL6,IL8,TGF-β,and IFNγ were higher(t=2.94,3.38,3.15,3.64,2.97;P=0.015,0.000,0.000,0.000,0.000)and the mRNA level of VEGF-A was lower(t=2.31,P =0.20) in senescent GES-1 cells than in the control group.In the conditioned medium of senescent GES-1 cells,the levels of IL-1β,IL6,IL8,TGF-β1,and IFNγ were higher in the H2O2-induced group [(3.12±0.21)μg/L,(4.26±0.15)μg/L,(3.37±0.14)μg/L,(5.34±0.19)μg/L,and(2.90±0.47)μg/L]than in the negative control group[(0.24±0.04,0.04±0.07,0.52±0.02,1.05±0.10,0.52±0.02,respectively,P＜0.001)],while the level of VEGF was lower in the H2O2-induced group than in the negative control group(0.21±0.03)μg/L vs (0.59±0.07)μg/L(P＜0.05).Conclusions The changes in senescence-associated secretory phenotype factors of the aging human gastric epithelial cells induced by oxidative stress may promote chronic gastritis and gastric cancer.

Objective To investigate the rescue effect of fixed position rescue team on emergency ectopic pregnancy patients.Methods A total of 42 ectopic pregnancy treated in our hospital from January to July 2017 were selected as observation group,given fixed position rescue.Another 40 ectopic pregnancy cases treated in our hospital from June to December 2016 were selected as control group,given routine emergency care.The treatment effect,the satisfaction of doctors and patients and the pressure of nursing staff were compared between the two groups.Results The observation group had shorter rescue time and the total rescue time,higher success rate of rescue,less hospital stay than the control group (P ＜ 0.05).The satisfaction of the patients and physicians in the observation group was higher,the physical and mental pressure and risk degree of the nursing staff were lower than that in the control group (P ＜ 0.05).Conclusion The fixed position rescue team can improve the rescue effect of emergency ectopic pregnancy patients,reduce the physical and mental pressure of nursing staff,and improve patients'satisfaction.

Objective To investigate the rescue effect of fixed position rescue team on emergency ectopic pregnancy patients.Methods A total of 42 ectopic pregnancy treated in our hospital from January to July 2017 were selected as observation group,given fixed position rescue.Another 40 ectopic pregnancy cases treated in our hospital from June to December 2016 were selected as control group,given routine emergency care.The treatment effect,the satisfaction of doctors and patients and the pressure of nursing staff were compared between the two groups.Results The observation group had shorter rescue time and the total rescue time,higher success rate of rescue,less hospital stay than the control group (P ＜ 0.05).The satisfaction of the patients and physicians in the observation group was higher,the physical and mental pressure and risk degree of the nursing staff were lower than that in the control group (P ＜ 0.05).Conclusion The fixed position rescue team can improve the rescue effect of emergency ectopic pregnancy patients,reduce the physical and mental pressure of nursing staff,and improve patients'satisfaction.