OBJECTIVE: This study investigated the effects of bis (2-butoxyethyl) phthalate (BBOP) on the onset of male puberty by affecting Leydig cell development in rats. METHODS: Thirty 35-day-old male Sprague-Dawley rats were randomly allocated to five groups mg/kg bw per day that were gavaged for 21 days with BBOP at 0, 10, 100, 250, or 500 mg/kg bw per day. The hormone profiles; Leydig cell morphological metrics; mRNA and protein levels; oxidative stress; and AKT, mTOR, ERK1/2, and GSK3β pathways were assessed. RESULTS: BBOP at 250 and/or 500 mg/kg bw per day decreased serum testosterone, luteinizing hormone, and follicle-stimulating hormone levels mg/kg bw per day (P < 0.05). BBOP at 500 mg/kg bw per day decreased Leydig cell number mg/kg bw per day and downregulated Cyp11a1, Insl3, Hsd11b1, and Dhh in the testes, and Lhb and Fshb mRNAs in the pituitary gland (P < 0.05). The malondialdehyde content in the testis significantly increased, while Sod1 and Sod2 mRNAs were markedly down-regulated, by BBOP treatment at 250-500 mg/kg bw per day (P < 0.05). Furthermore, BBOP at 500 mg/kg bw per day decreased AKT1/AKT2, mTOR, and ERK1/2 phosphorylation, and GSK3β and SIRT1 levels mg/kg bw per day (P < 0.05). Finally, BBOP at 100 or 500 μmol/L induced ROS and apoptosis in Leydig cells after 24 h of treatment in vitro (P < 0.05). CONCLUSION: BBOP delays puberty onset by increasing oxidative stress and apoptosis in Leydig cells in rats. UNLABELLED The graphical abstract is available on the website www.besjournal.com.
Rats ; Male ; Animals ; Leydig Cells/metabolism* ; Testosterone ; Glycogen Synthase Kinase 3 beta/pharmacology* ; Rats, Sprague-Dawley ; Sexual Maturation ; Testis ; Oxidative Stress ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis

This study aims to investigate the effect of Astragali Radix-Curcumae Rhizoma(AC) combination on the proliferation, migration, and invasion of colon cancer HT-29 cells based on epithelial-mesenchymal transition(EMT). HT-29 cells were respectively treated with 0, 3, 6 and 12 g·kg~(-1) AC-containing serum for 48 h. The survival and growth of cells were measured by thiazole blue(MTT) colorimetry, and the proliferation, migration, and invasion of cells were detected by 5-ethynyl-2'-deoxyuridine(EdU) test and Transwell assay. Cell apoptosis was examined by flow cytometry. The BALB/c nude mouse model of subcutaneous colon cancer xenograft was established, and then model mice were classified into blank control group, 6 g·kg~(-1) AC group, and 12 g·kg~(-1) AC group. The tumor weight and volume of mice were recorded, and the histopathological morphology of the tumor was observed based on hematoxylin-eosin(HE) staining. The expression of apoptosis-associated proteins B-cell lymphoma-2-associated X protein(Bax), cysteine-aspartic acid protease-3(caspase-3), and cleaved caspase-3, and EMT-associated proteins E-cadherin, MMP9, MMP2 and vimentin in HT-29 cells and mouse tumor tissues after the treatment of AC was determined by Western blot. The results showed that cell survival rate and the number of cells at proliferation stage decreased compared with those in the blank control group. The number of migrating and invading cells reduced and the number of apoptotic cells increased in the administration groups compared with those in the blank control group. As for the in vivo experiment, compared with the blank control group, the administration groups had small tumors with low mass and shrinkage of cells and karyopycnosis in the tumor tissue, indicating that the AC combination may improve EMT. In addition, the expression of Bcl2 and E-cadherin increased and the expression of Bax, caspase-3, cleaved caspase-3, MMP9, MMP2, and vimentin decreased in HT-29 cells and tumor tissues in each administration group. In summary, the AC combination can significantly inhibit the proliferation, invasion, migration, and EMT of HT-29 cells in vivo and in vitro and promote the apoptosis of colon cancer cells.
Humans ; Animals ; Mice ; Caspase 3 ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Vimentin ; HT29 Cells ; bcl-2-Associated X Protein ; Colonic Neoplasms ; Cell Proliferation

Objective: To study the specific killing effect of CD4 membrane protein targeted chimeric antigen receptor modified T (CAR-T) cell. Methods: The second generation CD4 targeted chimeric antigen receptor containing 4-1BB costimulation domain was insert into lentiviral vector through recombinant DNA technology. Lentivirus was prepared and packaged by 293T cells with four plasmids. Beads activated T cells were transduced with lentivirus and the transduction efficiency was checked with Protein L and flow cytometry. T cell subsets and IFN-γ concentrations were detected with probe-tagged antibody and cytometric bead assay. Results: ①The transduction efficiency of activated T cells with prepared lentivirus were 50.0%-70.0%. A subset of CD8⁺ T cell acquired dim expression of CD4 membrane protein after activation. CD4⁺T cell and CD8⁺CD4^dim T cell were gradually killed by CD4 targeted CAR-T post lentivirus transduction. ②The kill efficacy of CD4 targeted CAR-T cell and control T cell toward KARPAS 299 T cell at an E∶T ratio of 8∶1 for 24 h was (96.9±2.1)% and (11.2±3.1)%, CAR-T cell has a higher killing efficacy than control T cell (t=7.137, P=0.028). The IFN-γ concentrations in culture supernatant of CAR-T cell with K562-CD4 cell, CAR-T cell with K562 cell and CAR-T cell alone were (15 648±2 168), (1 978±354) and (1 785±268) pg/ml, CAR-T cell cocultured with K562-CD4 cell produced more IFN-γ than the other two controls (P<0.01). Conclusions: CD4 targeted CAR-T has an immunophenotype of CD8⁺CD4^-T cell. CD4 targeted CAR-T cell has killing efficacy toward normal CD4⁺T cell and CD4⁺T lymphoma cell. CD4 targeted CAR-T cell also has a killing efficacy toward CD4^dim target cell.
CD4 Antigens ; CD8-Positive T-Lymphocytes ; Cell Line, Tumor ; Humans ; Lymphoma ; Receptors, Antigen, T-Cell ; Receptors, Chimeric Antigen

Objective: To explore the efficacy and safety of haploidentical hematopoietic stem cell transplantation (Haplo-HSCT) for paroxysmal nocturnal hemoglobinuria (PNH). Methods: A total of 17 patients with PNH who received Haplo-HSCT from January 2013 to September 2017 were analyzed retrospectively. Results: Of them, 4 patients had de novo PNH, 13 patients had aplastic anemia-PNH syndrome (AA-PNH). All patients received modified busulfan and Cytoxan (BuCy)-based regimens combined with anti-thymocyte globulin (ATG). Granulocyte colony-stimulating factor-mobilized bone marrow and peripheral blood stem cells were transplanted as graft. Prophylaxis for graft-versus-host disease (GVHD) was ciclosporin A+ mycophenolate mofetil (MMF)+short-term methotrexate (MTX). All patients were engrafted successfully. The median time of neutrophils to 0.5×10(9)/L and platelets to 20×10(9)/L was 12(10-15) days and 14(11-45) days, respectively. All of the 17 patients achieved full donor chimerism at 30 d after Haplo-HSCT. Seven patients developed grade Ⅱ-Ⅳ acute GVHD, and 4 chronic GVHD. Median follow-up time was 27.1 (8.6-60.4) months. Of the 17 patients, 15 survived and 2 died of severe pulmonary infection and transplant associated thrombotic microangiopathy. Three-year overall survival was (77.8±15.2)%. Conclusion: Haplo-HSCT may be effective and safe for PNH patients who did not have matched donor.
Anemia, Aplastic ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Hemoglobins ; Humans ; Retrospective Studies ; Transplantation Conditioning

BACKGROUNDPatients on hemodialysis have a high-mortality risk. This study analyzed factors associated with death in patients on maintenance hemodialysis (MHD). While some studies used baseline data of MHD patients, this study used the most recent data obtained from patients just prior to either a primary endpoint or the end of the study period to find the characteristics of patients preceding death.

METHODSParticipants were selected from 16 blood purification centers in China from January 2012 to December 2014. Patients' data were collected retrospectively. Based on survival status, the participants were divided into two groups: survival group and the death group. Logistic regression analysis was performed to determine factors associated with all-cause mortality.

RESULTSIn total, 4104 patients (57.58% male, median age 59 years) were included. Compared with the survival group, the death group had more men and more patients with diabetic nephropathy (DN) and hypertensive nephropathy. The patients preceding death also had lower levels of diastolic blood pressure, hemoglobin, serum albumin, serum calcium, serum phosphate, Kt/V, and higher age. Multivariate analysis revealed that male sex (odd ratio [OR]: 1.437, 95% confidence interval [CI]: 1.094-1.886), age (OR: 1.046, 95% CI: 1.036-1.057), and presence of DN (OR: 1.837, 95% CI: 1.322-2.552) were the risk factors associated with mortality. High serum calcium (OR: 0.585, 95% CI: 0.346-0.989), hemoglobin (OR: 0.974, 95% CI: 0.967-0.981), albumin (OR: 0.939, 95% CI: 0.915-0.963) levels, and dialysis with noncuffed catheter (OR: 0.165, 95% CI: 0.070-0.386) were protective factors based on a multivariate analysis.

CONCLUSIONSHemodialysis patients preceding death had lower hemoglobin, albumin, and serum calcium levels. Multivariate analysis showed that male sex, age, DN, low hemoglobin, low albumin, and low serum calcium were associated with death in hemodialysis patients.

Adult ; Aged ; China ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Renal Dialysis ; adverse effects ; mortality ; Retrospective Studies ; Risk Factors

BACKGROUNDThe prevalence of developmental dysplasia of the hip (DDH) is unknown in China. We aimed to determine the prevalence of DDH in Chinese adults.

METHODSIn this study, we performed a cross-sectional survey of a nationally representative sample of Chinese adults. All participants underwent questionnaire investigation, physical examination, and X-ray examination. Factors associated with DDH were analyzed with logistic regression.

RESULTSWe invited 29,180 individuals aged 18 years and over to participate, randomly selected from 18 primary sampling units (street districts in urban areas and townships in rural areas). The survey and examination were completed in 25,767 people (10,296 men and 15,471 women). DDH was diagnosed in 391 people, yielding an overall DDH prevalence of 1.52%. Based on this information, we estimate the number of individuals with DDH in China to be approximately 16.05 million. DDH prevalence increased with age (odds ratio = 1.53 [1.03-2.27], P = 0.036), was significantly higher among women than men (2.07% vs. 0.75%, P< 0.001), and was higher among rural residents than urban residents (1.75% vs. 1.29%, P< 0.001). Economic development was independently associated with the presence of DDH. There was no evidence of an association between body mass index alone, education, or current smoking or drinking and risk of DDH (P > 0.05).

CONCLUSIONSDDH has become an important public health problem. Special attention should be paid to residents with DDH. Screening for DDH should be performed in China.

Adult ; Age Distribution ; Aged ; Body Mass Index ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Hip Dislocation, Congenital ; epidemiology ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Prevalence ; Risk Factors ; Sex Distribution

Objective: To explore the risk predictive value of lipoprotein-associated phospholipase A2 (Lp-PLA2) on acute coronary syndrome(ACS) and to study the relationship between Lp-PLA2 and the severity of coronary stenosis in ACS patients. Methods:A total of 155 ACS patients admitted in our hospital were enrolled. The patient were divided into 2 groups:AMI (acute myocardial infarction) group, n=49 and UA (unstable angina)group, n=106; in addition, there was a Control group, n=44 subjects with normal coronary angiography (CAG).Blood levels of Lp-PLA2 were examined, CAG was conducted and GRACE score, SYNTAX score,Gensini score were calculated. Based on Grace score, ACS patients were divided into 3 subgroups: Low risk subgroup, Grace score≤108, Mid risk subgroup,Grace score 109-140 and High risk subgroup,Grace score≥140.The above parameters were comparedamong different groups. Results: Compared with UA group and Control group, AMI group had increased blood level of Lp-PLA2, P<0.05. Compared with Low risk subgroup, High risk subgroup had much higher Lp-PLA2, P<0.05. Correlation analysis showed that Lp-PLA2 level was positively related to Gracescore (r=0.301, P<0.001). By SYNTAX score and Gensini score evaluation,Lp-PLA2 levels were similar among different subgroups. Conclusion:Blood level of Lp-PLA2 had certain risk predictive value in ACS patients; while it was not related to the severity of coronary stenosis.

Objective To understand the forming cause of the Oncomelania hupensis snail-existent non-endemic areas of schistosomiasis(SENEAS),and to verify the conclusion of previous studies,so as to provide the evidence for schistosomiasis monitoring in such areas in Nantong City,Jiangsu Province. Methods The controlled field tests were carried out to observe the O. hupensis snails artificially infected by schistosome miracidia in SENEAS. The influence of the soil from SENEAS and the en-demic areas on O. hupensis snails artificially infected by miracidia were observed. Results All the experimental snails could be infected by schistosome miracidia except the smooth-shell snails from Tangyuan Village in the controlled field test environment of SENEAS or the endemic areas. The infection rates of the smooth-shell snails were lower than those of the ribbed-shell snails , but there were no statistically significant differences. The mortality rates of the smooth-shell snails were higher than those of the ribbed-shell snails,which were statistically significant (χ2Xindian = 135.118,χ2Shuangdian = 122.836,χ2Baipu =154.436,χ2Dingyan =138.288,χ2Control=151.923,all P<0.01). There were no significant differences in the infection rates of snails between each test group of the soil from SENEAS and the endemic areas(χ2Rugao=0.071,χ2Rudong=0.216,both P>0.05). Also there was no signifi-cant difference between each test group and the control group without soil(χ2=7.148,P>0.05). Conclusion It is likely to form the spread of schistosomiasis in SENEAS in Nantong City with sufficient amount of infection source of schistosomiasis im-ported. It is still necessary to implement the surveillance of schistosomiasis and O. hupensis snails in Nantong City.

Objective:To retrospectively analyze the magnetic resonance imaging (MRI) signal differences between prostate cancer,benign prostatic hyperplasia,and normal prostate,and to explore the value of apparent diffusion coefficient (ADC) in prostate cancer grading.Methods:Forty cases of prostate cancer and eleven cases of benign prostatic hyperplasia in Zhongshan Hospital Affiliated to Fudan University between January 2013 to March 2014 were enrolled,which were all confirmed by surgery or biopsy.At the same time,20 normal cases were arranged as the control group,which were healthy volunteers.All the patients and volunteers were obtained informed consent.The T2WI signal intensity,T1 WI signal intensity (including unenhanced,arterial and venous phase) and ADC value of the three groups were compared after MRI and magnetic resonance diffusion weighted imaging (DWI).Then the correlation between ADC value of prostate cancer and Gleason score was analyzed to determine the critical value of ADC in the identification of high-risk and low-risk prostate cancer.Results:In terms of T2WI signal intensity and ADC values,there were significant differences among the three groups (P＜0.05).In terms of T1 WI signal intensity,only the difference of dynamic and venous phase in central gland of normal prostate was significantly different from that in prostate cancer and benign prostatic hyperplasia (P＜0.05),with the former higher than the latter two.The ADC value of prostate cancer was negatively correlated with Gleason score (r=-0.401,P=0.010).The higher the Gleason score,the smaller the ADC value.When the critical value of ADC was 0.70 × 10-3 rm2/s,the sensitivity of identifying low/intermediate-risk group and high-risk group for prostate cancer was 96.4％,and the specificity was 58.3％.Conclusions:T2WI signal intensity and ADC value can be used to identify prostate cancer,benign prostatic hyperplasia,and normal prostate.T2WI signal differences of dynamic and venous phase also have a certain reference value.The ADC value of prostate cancer is related to Gleason score,suggesting that it may be used for grading the clinical risk of prostate cancer.

OBJECTIVETo explore the relationship between serum creatinine (Scr) reference values in healthy adults and geographic factors and provide evidence for establishing Scr reference values in different regions.

METHODSWe collected 29 697 Scr reference values from healthy adults measured by 347 medical facilities from 23 provinces, 4 municipalities and 5 autonomous regions. We chose 23 geographical factors and analyzed their correlation with Scr reference values to identify the factors correlated significantly with Scr reference values. According to the Principal component analysis and Ridge regression analysis, two predictive models were constructed and the optimal model was chosen after comparison of the two model's fitting degree of predicted results and measured results. The distribution map of Scr reference values was drawn using the Kriging interpolation method.

RESULTSSeven geographic factors, including latitude, annual sunshine duration, annual average temperature, annual average relative humidity, annual precipitation, annual temperature range and topsoil (silt) cation exchange capacity were found to correlate significantly with Scr reference values. The overall distribution of Scr reference values featured a pattern that the values were high in the south and low in the north, varying consistently with the latitude change.

CONCLUSIONThe data of the geographic factors in a given region allows the prediction of the Scr values in healthy adults. Analysis of these geographical factors can facilitate the determination of the reference values specific to a region to improve the accuracy for clinical diagnoses.