BACKGROUND:Nerve growth factor is a protein that induces nerve growth and regulates biological behaviors such as proliferation and differentiation of mesenchymal stem cells. OBJECTIVE:To investigate the promoting effect of nerve growth factor on chondrogenic differentiation of bone marrow mesenchymal stem cells. METHODS:Rabbit bone marrow mesenchymal stem cells were isolated and cultured,and nerve growth factor was transfected into bone marrow mesenchymal stem cells by lentiviral transfection.The effects of nerve growth factor on the proliferation,migration,hypertrophic differentiation,and chondrogenic differentiation of bone marrow mesenchymal stem cells were detected by CCK-8 assay,cell scratch assay,alizarin red staining,and western blot assay,using the transfected null-loaded virus as control.To further investigate the promoting effect of nerve growth factor on the chondrogenic differentiation of bone marrow mesenchymal stem cells,interleukin 1β was added in bone marrow mesenchymal stem cells transfected with empty virus and nerve growth factor for 14 days.The expression of proteins related to chondrogenic differentiation and hypertrophic differentiation was detected by western blot assay. RESULTS AND CONCLUSION:(1)CCK-8 assay results showed that nerve growth factor had no significant effect on the proliferation of bone marrow mesenchymal stem cells.(2)Compared with the control group,overexpression of nerve growth factor enhanced the migration ability of the cells,and the expression of cartilage-associated proteins type II collagen and SOX9 was up-regulated(P<0.05),while the expression of hypertrophic-associated proteins type X collagen and Runx2 was down-regulated(P<0.05).(3)Compared with the empty virus+interleukin 1β group,the expression of cartilage-associated proteins type II collagen and Sox9 was up-regulated(P<0.05),and the expression of hypertrophy-associated proteins type X collagen and Runx2 was down-regulated after overexpression of nerve growth factor(P<0.05).(4)The results indicated that nerve growth factor could promote the chondrogenic differentiation of bone marrow mesenchymal stem cells.

Objective To evaluate the effectiveness of thermal tomography in breast cancer (BC) screening. Methods We conducted a general population-based BC screening in three regions of Hubei Province (Xiantao, Hongan, and Yangxin Districts). Participants underwent a questionnaire-based interview for baseline data collection. They then received a physical examination, thermal tomography, and ultrasound from doctors and technicians. We compared the efficacies, including sensitivity, specificity, and false-positive rates, of ultrasound and thermal tomography in BC screening. Results A total of 59 712 eligible women were included in this screening program. The BI-RADS 1, 2, 3, 4, and 5 accordance rates between the two screening methods were 0.9549, 0.8047, 0.9037, 0.3352, and 0, respectively. The overall accordance rate was 0.933 1. The kappa consistency results showed that the Cicchetti-Allison and Fleiss-Cohen kappa values were 0.7970 and 0.8583, respectively. Although the two methods had equal sensitivities, specificity was higher in thermal tomography than in ultrasound. The false-positive rate was lower in thermal tomography than in ultrasound. The area under the receiver operating characteristic (ROC) curve of thermal tomography was significantly larger than that of ultrasound. Conclusion The general consistency between thermal tomography and ultrasound in BC screening was high. Thermal tomography outperformed ultrasound in terms of specificity and diagnostic efficiency. Therefore, thermal tomography has a high application value for general population-based BC screening.

Nonalcoholic fatty liver disease （NAFLD） is an abnormal lipid metabolic disorder of the liver characterized by accumulation of a large amount of lipids in the liver， and it is currently the most common liver disease around the world. Mendelian randomization （MR） incorporates genomic data into traditional epidemiological study designs to infer the causal relationship between exposure factors and disease risk. In recent years， MR has been widely used in studies on inference of the etiology of NAFLD. This article systematically summarizes the advances in the application of MR in NAFLD research， so as to provide new ideas for understanding the nature of the disease and scientific interventions.

Objective To explore the relationship between serum soluble semaphorin 4D(sSema4D),CXC chemokine ligand 12(CXCL12)levels and left ventricular diastolic function in young and middle-aged patients with essential hypertension.Methods A total of 148 young and middle-aged patients with essential hyperten-sion admitted to a hospital from November 2020 to November 2022 were selected as the study subjects,and were grouped into left ventricular diastolic dysfunction group(n=41)and normal left ventricular diastolic function group(n=107)according to their left ventricular diastolic function.The serum levels of sSema4D and CXCL12 were detected by enzyme-linked immunosorbent assay.Pearson correlation analysis was applied to analyze the correlation between the serum levels of sSema4D and CXCL12 and the left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD),left ventricular septal thickness(IVST),left ventricular end-diastolic posterior wall thickness(LVPWT),left ventricular ejection fraction(LVEF),E peak/A peak(E/A)and maximum velocity of tricuspid regurgitation(TRVmax).The predictive value of ser-um sSema4D and CXCL12 levels in left ventricular diastolic dysfunction in young and middle-aged patients with essential hypertension was analyzed by receiver operating characteristic(ROC)curve.Results There were significant differences in diastolic blood pressure and gender between the left ventricular diastolic dys-function group and the left ventricular diastolic function normal group(P＜0.05).Compared with the normal left ventricular diastolic function group,serum levels of sSema4D,CXCL12 in the left ventricular diastolic dys-function group were obviously increased,and the difference was statistically significant(P＜0.05).Compared with normal left ventricular diastolic function group,IVST and LVPWT in the left ventricular diastolic dys-function group were significantly increased,and E/A was significantly decreased,with statistical significance(P＜0.05).Pearson correlation analysis showed that serum sSema4D and CXCL12 levels were positively cor-related with LVEDD,IVST and LVPWT(P＜0.05),and negatively correlated with E/A(P＜0.05).ROC curve analysis showed that the area under the curve(AUC)of serum sSema4D and CXCL12 combined in pre-dicting left ventricular diastolic dysfunction in young and middle-aged patients with essential hypertension was 0.894(95％CI:0.833-0.939),which was significantly greater than that of sSema4D alone in predicting left ventricular diastolic dysfunction in young and middle-aged patients with essential hypertension(Z=3.142,P=0.002)and CXCL12 alone predicted the AUC of left ventricular diastolic dysfunction in young and middle-aged patients with essential hypertension(Z=3.268,P=0.001).Conclusion Serum sSema4D and CXCL12 levels are associated with left ventricular diastolic function in young and middle-aged patients with essential hypertension.

Objective:To investigate the prognostic value of baseline peripheral blood inflammatory biomarkers for prognosis in patients with advanced hepatocellular carcinoma (HCC) receiving immunotherapy combined with targeted therapy.Methods:The clinical data of a total of 120 patients with advanced HCC who received immunotherapy combined with targeted therapy at Cancer Center of Renmin Hospital of Wuhan University from December 2019 to March 2022 were analyzed retrospectively. Receiver operating characteristic (ROC) curve was used to calculate the optimal cut-off values of neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), systemic immune inflammation index (SII) and prognostic nutritional index (PNI). According to the optimal cut-off values, the study objects were divided into high value group and low value group. The Kaplan-Meier method was used for survival analysis. Cox proportional hazard regression model was applied to analyze the factors associated with prognosis.Results:By the end of follow-up, 74 patients died and 46 survived. The median follow-up time was 23.0 months, the median overall survival (mOS) was 15.6 months, and the median progression-free survival (mPFS) was 13.1 months. ROC curve analysis showed that the optimal cut-off values of NLR, PLR, SII, LMR and PNI were 3.45, 131.87, 626.21, 2.12 and 43.30, respectively. The mPFS (18.3 months vs. 8.7 months) and mOS (26.6 months vs. 10.9 months) of patients in the low-NLR group ( n=75) were longer than those of the high-NLR group ( n=45), and there were statistically significant differences ( χ2=55.64, P<0.001; χ2=64.14, P<0.001). The mPFS (17.9 months vs. 10.9 months) and mOS (24.5 months vs. 13.5 months) of patients in the low-PLR group ( n=55) were longer than those of the high-PLR group ( n=65), and there were statistically significant differences ( χ2=5.27, P=0.023; χ2=11.84, P<0.001). The mPFS (18.0 months vs. 10.7 months) and mOS (25.7 months vs. 12.8 months) of patients in the low-SII group ( n=75) were longer than those of the high-SII group ( n=45), and there were statistically significant differences ( χ2=24.46, P<0.001; χ2=25.42, P<0.001). The mPFS (18.2 months vs. 10.9 months) and mOS (26.6 months vs. 13.2 months) of patients in the high-LMR group ( n=56) were longer than those of the low-LMR group ( n=64), and there were statistically significant differences ( χ2=19.25, P<0.001; χ2=19.92, P<0.001). The mPFS (17.9 months vs. 10.9 months) and mOS (25.4 months vs. 13.4 months) of patients in the high-PNI group ( n=62) were longer than those of the low-PNI group ( n=58), and there were statistically significant differences ( χ2=13.69, P<0.001; χ2=19.07, P<0.001). Univariate analysis showed that Barcelona clinic liver cancer (BCLC) stage ( HR=1.83, 95% CI: 1.17-2.87, P=0.008), Child-Pugh grade ( HR=2.21, 95% CI: 1.47-3.34, P<0.001), modified albumin-bilirubin (mALBI) grade ( HR=1.35, 95% CI: 1.01-1.81, P=0.045), extrahepatic metastases ( HR=2.18, 95% CI: 1.47-3.25, P<0.001), NLR ( HR=1.40, 95% CI: 1.28-1.54, P<0.001), PLR ( HR=1.00, 95% CI: 1.00-1.01, P=0.001), SII ( HR=1.00, 95% CI: 1.00-1.00, P<0.001), LMR ( HR=0.64, 95% CI: 0.51-0.79, P<0.001) and PNI ( HR=0.95, 95% CI: 0.93-0.98, P=0.001) were correlated with PFS; BCLC stage ( HR=2.18, 95% CI: 1.21-3.91, P=0.009), Child-Pugh grade ( HR=2.57, 95% CI: 1.61-4.09, P<0.001), Eastern Cooperative Oncology Group performance status score ( HR=1.59, 95% CI: 1.01-2.51, P=0.044), mALBI grade ( HR=1.60, 95% CI: 1.17-2.17, P=0.003), extrahepatic metastasis ( HR=2.51, 95% CI: 1.59-3.96, P<0.001), NLR ( HR=1.45, 95% CI: 1.32-1.60, P<0.001), PLR ( HR=1.01, 95% CI: 1.01-1.01, P<0.001), SII ( HR=1.01, 95% CI: 1.01-1.01, P<0.001), LMR ( HR=0.57, 95% CI: 0.40-0.72, P<0.001) and PNI ( HR=0.92, 95% CI: 0.89-0.96, P<0.001) were correlated with OS. Multivariate analysis showed that extrahepatic metastasis ( HR=1.78, 95% CI: 1.10-2.87, P=0.018) and NLR ( HR=1.46, 95% CI: 1.24-1.73, P<0.001) were independent influencing factors for PFS; extrahepatic metastasis ( HR=2.09, 95% CI: 1.21-3.61, P=0.009), NLR ( HR=1.56, 95% CI: 1.29-1.88, P<0.001), SII ( HR=1.00, 95% CI: 1.00-1.00, P=0.025), LMR ( HR=0.59, 95% CI: 0.45-0.78, P=0.008) and PNI ( HR=0.93, 95% CI: 0.88-0.99, P=0.013) were independent influencing factors for OS. Conclusion:NLR and extrahepatic metastasis can be regarded as important indicators to predict PFS in patients with advanced HCC receiving immunotherapy combined with targeted therapy, and NLR, SII, LMR, PNI and extrahepatic metastasis can be regarded as important indicators to predict OS in patients with advanced HCC receiving immunotherapy combined with targeted therapy. High NLR, high SII, low LMR, low PNI and extrahepatic metastasis indicate poor prognosis of HCC patients.

Objective To study the effect of mitochondrial reactive oxygen species(ROS)on male fertility and its application value in assisted reproduction so as to provide a new method for fertility assessment and etiological treatment of male infertility.Methods The relationship between sperm ROS and male infertility,sperm nuclear DNA integrity and sperm membrane function was analyzed.Then the relationships of the ROS of sperms for fertiliza-tion with fertilization rate,cleavage rate,embryo rate and pregnancy outcome were analyzed.Results(1)The percentage of the high ROS in sperm membrane of the male infertility group was significantly higher than that of the normal fertility group(P<0.01),but there was no statistical difference in the percentage of the high ROS in the sperm mitochondria between the two groups.(2)The normal rate of sperm membrane function in the normal group was significantly higher than that in the abnormal group(P<0.01),but there was no significant difference in the sperm DNA fragmentation index(DFI)between the groups.(3)There was significant difference in the fertilization rate between the IVF control group and IVF observation group,but no significant difference was seen in the cleavage rate,excellent embryo rate and clinical pregnancy rate between the groups.Conclusion Sperm ROS is related to male fertility to some extent.Abnormal ROS may lead to impaired sperm membrane function and affect sperm fertilization ability,thus affecting male fertility.The detection of ROS in sperm can be used as a new method to evaluate male fertility and provide a basis for diagnosis or treatment of male infertility.For patients with ART or infertility,the appropriate time to conceive can be selected according to the sperm ROS level.

【Objective】 To investigate the sperm retrieval rate (SRR) of microdissection testicular sperm extraction (M-TESE) in patients with non-obstructive azoospermia (NOA) caused by different causes. 【Methods】 A retrospective analysis was performed on 225 NOA patients during Jan.2020 and Dec.2022. The relation between SRR and patients’ age,body mass index (BMI),testicular volume,endocrine hormones and different etiological classifications were analyzed. 【Results】 According to whether sperm was obtained by surgery,the patients were divided into two groups,including 107 cases in the sperm group and 118 cases in the non-sperm group. There were no significant differences in patients’ age,testicular volume and levels of endocrine hormones between the two groups (P>0.05). According to the different causes,NOA patients with mumps history,cryptorchidism history,AZFc deletion or Klinefelter syndrome (KS) had higher SRR,while idiopathic NOA patients had the lowest SRR (P<0.05). 【Conclusion】 M-TESE is an effective treatment of NOA. There is no correlation between SRR and patients’ age,MBI,testicular volume and levels of endocrine hormones. NOA caused by different etiological classifications may have different SRR.

【Objective】 To investigate the effects of the loss of exon 1 of TFE3 on nuclear localization of chimeric TFE3 protein in TFE3 translocation renal cell carcinoma (TFE3 tRCC). 【Methods】 The localization of TFE3 protein in TFE3 tRCC and clear cell renal cell carcinoma (ccRCC) were detected with immunochemistry. The exon retention of TFE3 gene in TFE3 tRCC was analyzed in databases and literatures. The plasmids containing TFE3 full-length and different-length of TFE3 exons which were constructed to pCDH-MCS-EGFP-Puro were transfected into HEK293T using Lipo Fiter^TM. The localization of EGFP protein in HEK293T cells were detected with confocal microscopy. The localization of TFE3 protein and truncated TFE3 protein were detected with Western blotting. The mRNA expression of the downstream genes of TFE3 protein were detected with q-PCR. 【Results】 Strong nuclear signal of TFE3 protein was observed in TFE3 tRCC, whereas TFE3 protein in ccRCC was mainly localized in cytoplasm. The results of fluorescence imaging and Western blotting showed that TFE3 full-length protein was expressed both in nucleus and cytoplasm, and the expression of truncated TFE3 protein was mainly localized in nucleus. The q-PCR analysis demonstrated that the deletion of exon 1 in TFE3 gene led to a higher transcriptional level of targeted genes of TFE3 protein. 【Conclusion】 The loss of exon 1 in TFE3 played a critical role in preventing TFE3 protein from entering the nucleus. In TFE3 tRCC, the loss of exon 1 in TFE3 gene leads to the nuclear localization of TFE3 fusion protein and activation of its downstream target genes. This mechanism promises to uncover the occurrence and development of TFE3 tRCC.

Humans ; Hypertension, Pulmonary/surgery* ; Learning Curve ; Angioplasty, Balloon ; Pulmonary Embolism/therapy* ; Chronic Disease ; Pulmonary Artery/surgery*

Objective:To explore the value of pre-ablation stimulated thyroglobulin (psTg) before 131I treatment combined with lymph node ratio (LNR) in predicting 131I treatment response in patients with papillary thyroid cancer (PTC). Methods:From January 2016 to December 2018, 178 PTC patients (47 males, 131 females; age (43.2±12.6) years) treated with 131I in the Affiliated Cancer Hospital of Zhengzhou University were retrospectively analyzed. According to 131I treatment response, patients were divided into excellent response (ER) group and non-ER group. The clinical data of the two groups were compared by χ2 test, independent-sample t test and Mann-Whitney U test. The cut-off values and AUCs of psTg and LNR to predict treatment response were calculated according to the ROC curve. Factors affecting 131I treatment response were analyzed by logistic multivariate regression analysis. Results:There were 118 patients (66.3%, 118/178) in ER group and 60 patients (33.7%, 60/178) in non-ER group, and there were significant differences in N stage ( χ2=11.15, P=0.004), 131I treatment dose ( χ2=12.65, P<0.001), American Thyroid Association (ATA) initial risk stratification ( χ2=15.25, P<0.001), number of metastatic lymph nodes ( χ2=22.63, P<0.001), LNR ( U=1 506.00, P<0.001) and psTg ( U=919.00, P<0.001) between the two groups. The cut-off values of psTg and LNR predicting ER were 3.97 μg/L and 0.29, with the AUC of 0.870 and 0.787 respectively. PsTg (odds ratio ( OR)=10.88, 95% CI: 4.67-25.36, P<0.001) and LNR ( OR=5.30, 95% CI: 1.85-15.23, P=0.002) were independent factors to predict 131I treatment response in PTC patients. When psTg≥3.97 μg/L, LNR ( OR=9.40, 95% CI: 2.06-42.92, P=0.004) was an independent factor affecting 131I treatment response in PTC patients. Conclusions:PsTg and LNR are independent factors affecting 131I treatment response in PTC patients. When psTg≥3.97 μg/L, LNR can be used as a supplementary factor to predict 131I treatment response. The combination of psTg and LNR can better predict 131I treatment response in PTC patients.