Objective To establish a risk model of rivaroxaban-related bleeding events in elderly patients with non-valvular atrial fibrillation(NVAF)based on gene polymorphism.Methods A total of 268 elderly NVAF patients receiving rivaroxaban treatment in Department of General Practice of the First Affiliated Hospital of Shihezi University from January 2021 to July 2023 were enrolled in this study.According to whether bleeding events occurred in 12 months'follow-up,they were divided into a bleeding group(47 cases)and a non-bleeding group(221 cases).The clinical data and results of gene polymorphism were compared between the two groups.Multivari-ate logistic regression was adopted to construct a risk prediction model of bleeding events based on gene polymorphism,and the predictive performance was verified.Results Significantly ad-vanced age and lower creatinine level were observed in the bleeding group than the non-bleeding group(P＜0.01).The bleeding group had obviously lower GG genotype frequency at the rs1128503 locus and TT genotype frequency at the rs4148738 of ATP-binding cassette,sub-family B member 1(ABCB1),and lower AC genotype frequency at the rs1057910 locus of cytochrome P450 2C9(CYP2C9)than the non-bleeding group(P＜0.05).Multivariate logistic regression analy-sis showed that age(OR=1.136,95％CI:1.031-1.251),and AA genotype(OR=15.407,95％CI:4.259-55.741)and GA genotype(OR=6.990,95％CI:1.599-30.546)of ABCB1 rs1 128503 were risk factors for rivaroxaban-related bleeding events in elderly NVAF patients(P＜0.01).Creatinine(OR=0.943,95％CI:0.899-0.989),TT genotype at ABCB1 rs4148738(OR=0.048,95％CI:0.009-0.242)and AC genotype of CYP2C9 rs1057910(OR=0.092,95％CI:0.021-0.408)were protective factors for the risk(P＜0.05).ROC curve analysis indicated that the AUC value of the risk model based on gene polymorphism was 0.827(95％CI:0.776-0.870),the spe-cificity was 81.90％,and the sensitivity was 76.60％.Conclusion Our risk model of rivaroxaban-related bleeding events based on ABCB1 gene and CYP2C9 gene has high application value for elderly NVAF patients.

Objective To establish a risk model of rivaroxaban-related bleeding events in elderly patients with non-valvular atrial fibrillation(NVAF)based on gene polymorphism.Methods A total of 268 elderly NVAF patients receiving rivaroxaban treatment in Department of General Practice of the First Affiliated Hospital of Shihezi University from January 2021 to July 2023 were enrolled in this study.According to whether bleeding events occurred in 12 months'follow-up,they were divided into a bleeding group(47 cases)and a non-bleeding group(221 cases).The clinical data and results of gene polymorphism were compared between the two groups.Multivari-ate logistic regression was adopted to construct a risk prediction model of bleeding events based on gene polymorphism,and the predictive performance was verified.Results Significantly ad-vanced age and lower creatinine level were observed in the bleeding group than the non-bleeding group(P＜0.01).The bleeding group had obviously lower GG genotype frequency at the rs1128503 locus and TT genotype frequency at the rs4148738 of ATP-binding cassette,sub-family B member 1(ABCB1),and lower AC genotype frequency at the rs1057910 locus of cytochrome P450 2C9(CYP2C9)than the non-bleeding group(P＜0.05).Multivariate logistic regression analy-sis showed that age(OR=1.136,95％CI:1.031-1.251),and AA genotype(OR=15.407,95％CI:4.259-55.741)and GA genotype(OR=6.990,95％CI:1.599-30.546)of ABCB1 rs1 128503 were risk factors for rivaroxaban-related bleeding events in elderly NVAF patients(P＜0.01).Creatinine(OR=0.943,95％CI:0.899-0.989),TT genotype at ABCB1 rs4148738(OR=0.048,95％CI:0.009-0.242)and AC genotype of CYP2C9 rs1057910(OR=0.092,95％CI:0.021-0.408)were protective factors for the risk(P＜0.05).ROC curve analysis indicated that the AUC value of the risk model based on gene polymorphism was 0.827(95％CI:0.776-0.870),the spe-cificity was 81.90％,and the sensitivity was 76.60％.Conclusion Our risk model of rivaroxaban-related bleeding events based on ABCB1 gene and CYP2C9 gene has high application value for elderly NVAF patients.

Objective:To explore whether left ventricular interstitial fibrosis is associated with left atrial enlargement and left atrial dysfunction in patients of hypertrophic cardiomyopathy(HCM) with preserved ejection fraction.Methods:From October 2018 to September 2021, 59 HCM including 30 with enlarged maximal left artrial volume index (LAVI max), 29 with normal LAVI max and 28 age-and gender-matched controls were retrospectively enrolled. Imaging protocol included cine sequence, late gadolinium enhancement and T 1 mapping.The relationships between left ventricular mass index (LVMI), quantitative myocardial fibrosis and left atrial-related indexes were analyzed. One-way analysis of variance with Bonferroni post hoc correction or Kruskal-Wallis was performed for continuous variables. Categorical variables were assessed using the Chi-square test or Fisher′s exact test. Pearson or Spearman analysis was used for linear or monotonic nonlinear correlations. Results:The left ventricular end-diastolic volume index, left ventricular end-systolic volume index, left ventricular cardiac output and LVMI of HCM with enlarged LAVI max group were higher than HCM with normal LAVI max group and control group( P<0.05).Correlation analysis showed that LVMI correlated positively with LAVI max( r=0.780, P<0.001) and minimal left artrial volume index (LAVI min) ( r=0.816, P<0.001), extracellular volume correlated positively with LAVI max( r=0.462, P<0.001) and LAVI min( r=0.483, P<0.001),%LGE was correlated positively with LAVI max( r=0.311, P<0.05) and LAVI min( r=0.327, P<0.05),left ventricular index interstitial volume was correlated negatively with left atrial ejection fraction of reservoir ( r=-0.669, P<0.001),left atrial ejection fraction of conduit ( r=-0.472, P<0.001),left atrial ejection fraction of pump ( r=-0.518, P<0.001)and left atrial expansion index( r=-0.626, P<0.001). Conclusion:There is association between LVMI and fibrosis and left atrial enlargement and phases dysfunction in HCM with preserved ejection fraction.

Objective:To investigate the effects of fluoride exposure on proliferation, apoptosis and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in mice.Methods:BMSCs were isolated and cultured from femur bone marrow of C57BL/6 mice (6 - 8 weeks). The cells in passage 3 were used to detect the surface markers of stem cells by flow cytometry. The cells were cultured in media with a final fluoride concentration of 0.0, 0.1, 1.0, 5.0, 10.0, 15.0, 20.0 and 40.0 mg/L, respectively. The effects of different fluoride concentrations on BMSCs cell proliferation (CCK8 method), apoptosis (flow cytometry analysis), osteogenic differentiation ability [alizarin red and alkaline phosphatase (ALP) staining] were detected. Western blot was applied to detect the levels of apoptosis-related proteins [poly ADP-ribose polymerase (PARP)], mitogen-activated protein kinase (MAPK) pathway member proteins [extracellular regulated protein kinase 1/2 (ERK1/2), c-Jun amino-terminal kinase (JNK), p38 and phosphorylated ERK, JNK, p38 (p-ERK, p-JNK, p-p38)], osteogenic differentiation-related protein [Runt-related transcription factor 2 (Runx2), ALP] and Wnt/β-catenin pathway member proteins [glycogen synthase kinase-3β (GSK3β), phosphorylated GSK3β (p-GSK3β) and β-catenin]. Immunocytofluorescense staining was applied to evaluate the expression levels of p-GSK3β and β-catenin. The two pathways (MAPK and Wnt/β-catenin) were blocked by SP600125 and DKK-1, respectively, to testify their involvement in mechanisms of apoptosis and osteogenic differentiation.Results:The mouse BMSCs were successfully isolated and cultured. Flow cytometry analysis showed that the mesenchymal stem cell surface biomarkers (CD73, CD90 and CD105) were positively expressed. The comparison of cell proliferation at three time points (24, 48 and 72 h) in each concentration group was statistically significant ( F = 65.36, 160.04 and 365.32, P < 0.001), and the comparison of early apoptosis (24 h) in each concentration group was statistically significant ( F = 214.04, P < 0.001); compared with the 0.0 mg/L group, the cell proliferation in 15.0, 20.0 and 40.0 mg/L groups decreased, and the early apoptosis rate in 10.0, 15.0 and 20.0 mg/L groups increased ( P < 0.05). When cells were treated with 15.0 mg/L fluoride for 0 - 24 h, the p-JNK/JNK ratio was higher at 2, 4, 8, 12, 18 and 24 h compared with that at 0 min ( P < 0.05); compared with the fluoride group (15.0 mg/L), the early apoptosis rate of cells after SP600125 block decreased ( P < 0.05), and the protein expression levels of PARP and p-JNK decreased ( P < 0.05). After osteogenic induction, compared with the 0.0 mg/L group, in 0.1 and 1.0 mg/L groups ALP staining was enhanced and the number of calcified nodules increased, and the protein expression levels of Runx2 and ALP in the 0.1 and 1.0 mg/L groups were higher ( P < 0.05). After osteogenic induction, compared with the 0.0 mg/L group, the p-GSK3β/GSK3β ratio and β-catenin protein level were significantly higher in the 0.1 and 1.0 mg/L groups ( P < 0.05); and compared with the fluoride group (1.0 mg/L), addition of DKK-1 significantly decreased the protein expression levels of p-GSK3β and β-catenin and reduced the nuclear entry of β-catenin, and ALP staining decreased and the number of calcified nodules decreased. Conclusions:High concentration of fluoride (> 10.0 mg/L) inhibits the proliferation and promotes apoptosis of BMSCs, while low concentration of fluoride (0.1, 1.0 mg/L) promotes osteogenic differentiation. The MAPK/JNK pathway and the classical Wnt pathway are involved in the above cellular processes, respectively.

Objective:To explore the influencing factors of embryos quality during the cycle of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) and pregnancy outcomes of frozen-thawed embryo transfer (FET) in patients with polycystic ovary syndrome (PCOS). Methods:A retrospective case-control study design was used to analyze patients who received IVF/ICSI treatment at the Reproductive Medicine Center of Tianjin Central Obstetrics and Gynecology Hospital from January 2015 to December 2019, underwent whole embryo cryopreserved and performed the first FET. The 1233 cycles included were divided into control group ( n=561) and PCOS group ( n=672) according to PCOS diagnosis. The general clinical characteristics, laboratory-related indicators and pregnancy outcomes of patients between the two groups were compared, and the affecting factors of the late miscarriage rate were analyzed by multivariate logistic regression. Results:1) In terms of the general clinical characteristics between the two groups, the differences of duration of infertility [(3.95±2.01) years vs. (4.84±2.91) years, P=0.007], body mass index (BMI) [(21.96±2.52) kg/m 2vs. (23.96±3.50) kg/m 2, P<0.001], basal luteinizing hormone [(4.71±2.38) mU/L vs. (8.18±5.40) mU/L, P<0.001], basal estradiol [(45.49±31.80) ng/L vs. (56.67±54.17) ng/L, P=0.032], basal testosterone [(42.80±13.45) ng/L vs. (53.45±38.67) ng/L, P=0.001], gonadortopin initial used dosage [(230.80±54.07) U vs. (192.11±53.79) U, P<0.001] were statistically significant. The endometrium preparation plan in the FET cycle, more PCOS group patients received hormone replacement treatment [64.1% (431/672) vs. 26.6% (149/561)], while more patients in control group received natural cycle transplantation [73.4% (412/561) vs. 35.9% (241/672)], and the differences were statistically significant (all P<0.001). 2) In terms of the laboratory results, the number of oocytes retrieved in PCOS group (23.36±9.53) was higher than that in control group (20.32±8.81, P=0.002). The number of high-quality embryos and the rate of high-quality embryos in PCOS group [2.94±3.13; 33.3% (2016/6048)] were lower than those in control group [4.17±3.65, P=0.034; 46.3% (2339/5049), P<0.001], and the differences were statistically significant. 3) In the pregnancy outcomes, the high-quality embryo transfer rate and the biochemical pregnancy rate in control group were higher than those in PCOS group [71.0% (743/1046) vs. 59.3% (761/1284), P<0.001; 7.3% (41/561) vs. 4.5% (30/672), P=0.033], and the late miscarriage rate in PCOS group [10.3% (43/418)] was higher than that in control group [4.3% (16/326), P=0.002]. 4) Logistic regression analysis was performed on the influencing factors of late miscarriage. After correcting the confounding factors, PCOS ( OR=2.573, 95% CI=1.270-5.212, P=0.009) and maternal high BMI ( OR=1.080, 95% CI=0.991-1.176, P=0.031) were the risk factors for late miscarriage. Conclusion:The number of high-quality embryos and the rate of high-quality embryos in PCOS patients were lower than those in non-PCOS patients. PCOS and high BMI were risk factors for late miscarriage in patients. Improving endocrine disorders and weight control in PCOS patients before fertility treatment is of positive significance for improving the pregnancy outcome of patients.

Objective:To explore the influencing factors of embryos quality during the cycle of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) and pregnancy outcomes of frozen-thawed embryo transfer (FET) in patients with polycystic ovary syndrome (PCOS). Methods:A retrospective case-control study design was used to analyze patients who received IVF/ICSI treatment at the Reproductive Medicine Center of Tianjin Central Obstetrics and Gynecology Hospital from January 2015 to December 2019, underwent whole embryo cryopreserved and performed the first FET. The 1233 cycles included were divided into control group ( n=561) and PCOS group ( n=672) according to PCOS diagnosis. The general clinical characteristics, laboratory-related indicators and pregnancy outcomes of patients between the two groups were compared, and the affecting factors of the late miscarriage rate were analyzed by multivariate logistic regression. Results:1) In terms of the general clinical characteristics between the two groups, the differences of duration of infertility [(3.95±2.01) years vs. (4.84±2.91) years, P=0.007], body mass index (BMI) [(21.96±2.52) kg/m 2vs. (23.96±3.50) kg/m 2, P<0.001], basal luteinizing hormone [(4.71±2.38) mU/L vs. (8.18±5.40) mU/L, P<0.001], basal estradiol [(45.49±31.80) ng/L vs. (56.67±54.17) ng/L, P=0.032], basal testosterone [(42.80±13.45) ng/L vs. (53.45±38.67) ng/L, P=0.001], gonadortopin initial used dosage [(230.80±54.07) U vs. (192.11±53.79) U, P<0.001] were statistically significant. The endometrium preparation plan in the FET cycle, more PCOS group patients received hormone replacement treatment [64.1% (431/672) vs. 26.6% (149/561)], while more patients in control group received natural cycle transplantation [73.4% (412/561) vs. 35.9% (241/672)], and the differences were statistically significant (all P<0.001). 2) In terms of the laboratory results, the number of oocytes retrieved in PCOS group (23.36±9.53) was higher than that in control group (20.32±8.81, P=0.002). The number of high-quality embryos and the rate of high-quality embryos in PCOS group [2.94±3.13; 33.3% (2016/6048)] were lower than those in control group [4.17±3.65, P=0.034; 46.3% (2339/5049), P<0.001], and the differences were statistically significant. 3) In the pregnancy outcomes, the high-quality embryo transfer rate and the biochemical pregnancy rate in control group were higher than those in PCOS group [71.0% (743/1046) vs. 59.3% (761/1284), P<0.001; 7.3% (41/561) vs. 4.5% (30/672), P=0.033], and the late miscarriage rate in PCOS group [10.3% (43/418)] was higher than that in control group [4.3% (16/326), P=0.002]. 4) Logistic regression analysis was performed on the influencing factors of late miscarriage. After correcting the confounding factors, PCOS ( OR=2.573, 95% CI=1.270-5.212, P=0.009) and maternal high BMI ( OR=1.080, 95% CI=0.991-1.176, P=0.031) were the risk factors for late miscarriage. Conclusion:The number of high-quality embryos and the rate of high-quality embryos in PCOS patients were lower than those in non-PCOS patients. PCOS and high BMI were risk factors for late miscarriage in patients. Improving endocrine disorders and weight control in PCOS patients before fertility treatment is of positive significance for improving the pregnancy outcome of patients.

OBJECTIVE：To establish a method for the determination of aloesin in plasma of rats ，and to investigate pharmacokinetic characteristics of aloesin. METHODS ：The plasma samples were precipitated with methanol. Using aloeresin D as internal standard ，the plasma concentration of aloesin was determined by LC-MS/MS. The determination was performed on Synergi Hydro-RP column with mobile phase consisted of 0.1‰ formic acid-methanol （gradient elution ）at the flow rate of 0.50 mL/min. The column temperature was 30 ℃，and sample size was 5 µL. The electrospray ionization source was applied to carry out negative ion detection with multiple reaction monitoring mode . The ion transitions for quantitative analysis were m/z 393.1→272.9（aloesin） and m/z 555.3→144.9（internal standard ），respectively. The concentration of aloesin in venous blood was determined by above method at 0.083，0.167，0.333，0.667，1，1.5，2.5，4，6，8，10 h after intravenous injection （3.35 mg/kg）and intragastric administration（16.75 mg/kg）of aloesin. DAS 3.0 software was used to calculate pharmacokinetic parameters. RESULTS ：The linear range of aloesin were 1-600 ng/mL（r＝0.994 5）. The lower limit of quantification was 1 ng/mL，and RSDs of within and between batches were less than 15％；accuracies within and between batches were within ±15％. The matrix factors were （92.74± 4.33）％-（94.84±2.57）％，and extraction recoveries were （69.04±2.13）％-（75.03±2.84）％；the deviation between the measured results of the stability test and the theoretical values were within ±15％. After intravenous injection and intragastric administration of aloesin ，main pharmacokinetic parameters were as follows ：cmax were（10 693.3±2 745.3）and（223.3±36.2）ng/mL；t1/2 were （2.45±1.45）and（3.33±1.91）h；AUC0－24h were（4 190.6±883.6）and（1 210.1±93.9）ng·h/mL（n＝3）. Absolute bioavailabi- lity was 11.13％. CONCLUSIONS ：The established method is rapid and sensitive for plasma determination of aloesin ，and suitable for its pharmacokinetic study.

Objective ToexplorethevalueofMRIindifferentiatingbetweenmass-formingchronicpancreatitis(MFCP)andpancreatic carcinoma(PC).Methods MRIdataof19caseswith MFCP,36caseswithPCand30normalcontrolcasesconfirmedbypathology orfollow-upwereanalyzedretrospectively.AllofthesubjectsunderwentroutineMRIandDWIscan.MRIcharacteristicsofdiseases andnormalpancreaswereanalyzed,andADCvalueswerecomparedamongthethreegroups.Results TheaverageADCvalueofthe MFCPgroupwas(1.41±0.25)×10-3 mm2/s,higherthanthatofthePCgroup (1.13±0.11)×10-3 mm2/s(P<0.05),andlower thanthatofthenormalcontrolgroup(1.50±0.27)×10-3 mm2/s(P<0.05).IntheT2WI,enhancedscanningarterialphase,andenhanced scanningportalphase,thesignalcharacteristicsofthelesionswerestatisticallydifferentbetweentheMFCPandPCgroup (P<0.05).The sensitivityandspecificityofthecombinationT2WI,enhancedarterialimagingandADCvaluewere86.9%and88.9%indifferentiatingMFCPand PC,whichwasbetterthananysinglemethod.Conclusion MRImulti-sequencecombinationisoneoftheeffectivemethodsforidentifyingPCand MFCP,andhasreferencevalueforclinicaldiagnosis.

Objective To investigate the diagnostic value of multiparametric MRI in early prostate cancer(PCa) based on PI-RADS version 2.Methods 27 surgically-proved early PCa patients were collected in this retrospective study.T2WI,DWI and DCE were evaluated by two blinded radiologists.By 12 sub-region classification method the possibility of the presence of cancer at each sub-region was scored according to the PI-RADS V2.The receiver operating characteristic (ROC) curve was used to analyze the diagnosic efficacy of the following 4 protocols:T2WI alone(protocol 1),T2WI+DWI(protocol 2),T2WI+DCE(protocol 3),T2WI+DWI+DCE(protocol 4).The sensitivity,specificity and accuracy for each protocol were calculated.The average scores of cancerous sub-regions and non-cancerous sub-regions were calculated and the independent sample t test was used to compare the four protocols.Results 324 sub-regions were analyzed in 27 early PCa patients and then divided into 119 cancerous sub-regions and 205 non-cancerous sub-regions,including 64 peripheral zone cancerous sub-regions and transition zone cancerous sub-regions.In protocol 1-4, the average scores of cancerous sub-regions in orderwere 3.13±1.19,3.27±1.15,3.28±1.23, 3.33±1.16,respectively.Non-cancerous sub-regions's scores in order were 1.98±0.90,1.91±0.91, 2.03±0.99,1.94±0.96 respectively and there were significant differences among each protocol (P<0.05).The area under the ROC curve of the 4 protocols for region-based analysis were displayed in descending order: protocol 4 (0.819), protocol 2 (0.810), protocol 3 (0.772), protocol 1 (0.765) and there were no significant differences between any two protocols (P>0.05).In four protocols, the sensitivity in order were 45.40%, 56.30%, 59.70%, 61.34%, while the specificity in order were 95.10%, 96.10%, 89.80%, 96.60%, and the accuracy in order were 76.85%, 81.48%, 78.70%, 83.65%.Conclusion Multiparametric MRI can improve the diagnostic accuracy for the detection of early PCa, and T2WI+DWI+DCE is with the highest value.The PI-RADS V2 system is a better semi quantitative method for evaluation of early PCa.

OBJECTIVETo establish a method for isolation, cryopreservation and recovery of the highly viable human peripheral blood monomuclear cells (PBMNCs) so as to achieve the long-term preservation of PBMNCs.

METHODSA total of 80-100 ml peripheral blood were collected from the healthy volumteers aged over 50 years old. The PBMNCs were isolated by the Ficoll density gradient technique and cryopreserved gradually by program control method in liquid nitrogen freezer of -196 °C. The serum-free medium and autoloqous plasma medium were test for preservation of PBMNCs. The cell viability was assessed at time point of 1, 2, 4, 8, 12 and 24 months after thawing. Finally, the proliferation ability, purity and cytotoxicity were compared between the autologous immune lymphocytes (AIL) induced from cryopreserved PBMNCs and AIL as control from fresh PBMNCs.

RESULTSAfter separating, the cell viability was 99.6%±0.4%, and the recovery rate of lymphocytes was 58.4%±6.52%. The cell recovery rate of lymphocyte was 89.7%±3.82% at 24 months. The quality assurance program was reliable within 2 years of running. The AIL cells induced with cryopreserved PBMNCs were not significantly different from those induced from fresh PBMNCs in terms of proliferative action, purity and cytotoxicity(CD3(+)CD8(+) ≥45%,CD3(+)CD56(+) NKT≥10%,CD4(+)CD25(+) NKT≤10%).

CONCLUSIONManual separation of lymphocytes in vitro can get enough high-quality PBMNCs. The long-term cryopreserved PBMNC still maintain their high viability. The reinfusion of the clinical autologous immune cells would be advantageous for early tumor immunotherapy. Human AIL induced from cryopreserved PBMNC maintain their anti-tumor ability. These findings have the important implications for the application of these cells to adoptive cellular therapy.