Objective To study endocrine disruptor chemicals causing malformed frogs in an irrigation canal in T city and a pool in CH city. Methods Water samples collected from an irrigation canal in T city and a pool in CH city where malformed frogs were found were concentrated through H103 resin. The concentrated samples were purified and derivated. The endocrine disruptor chemicals in the derivatives were determined by GC/MS and the estrogenic activities in concentrated water samples were assessed by the recombinant yeast estrogen system (RYES). Results Endocrine disruptor chemicals bisphenol_A (5.6?10-3～1.52 ?g/L) and nonylphenol (0.46～0.58 ?g/L) were found in the concentrated water samples of the irrigation canal in T city and the pool in CH city. RYES showed that the estrogenic activities were positive and this result accorded with the data given by GC/MS. Conclusion The results indicated that the cause resulted in malformation of frogs in the irrigation canal in T city and the pool in CH city might be related to endocrine disruptor chemicals.

Objective To investigate the possibility of plastic and cosmetic operation incorporating epicanthoplasty with the double-eyelid procedure in one stage to achieve a more subtle scar in epicanthus patients with single eyelid. Methods After designing incision line of the epicanthic flap, double-eyelid line and the lower eyelid incision curved line, we cut the medial canthus skin transversely to reach a new medial canthus point, incised the part of front angle of medial palpebral ligamentum longitudinally and fixed the end point of front angle of medial palpebral ligamentum to nasal dorsum fascia firmly. Double eyelid operation was routinely performed to remove redundant small triangle skin as well as some patchy of orbicularis on the new medial canthus point. Along with eyelid edge about 1 to 2 mm to eyelash, the temporal side was cut with curved line till its disappearance by eyelid's "cat ear". The lower eyelid flap was separated downward and the superfluous flap and some patchy of orbicularis cut. The skin was su-tured to make postoperative scar hidden, blepharophimosis increased and the fold disappeared. Results 46 eyes (23 cases) were operated and satisfactory aesthetic results were obtained. Palpebral fissure was enlarged to 2 to 4 mm with epicanthic scar disappearance and formation of double-eyelid. Conclusions This is a simple and effective procedure with hidden epicanthic scar and the double-eyelid blepharoplasty could be performed simultaneously. Most patients receive satisfactory results during the 0.5 to 2 years' follow-up period except 2 cases with mild proliferation of epicanthus in half a year. It is especially suitable to correct the severe epicanthus palpebralis or epieanthus tarsalis.

A novel method for the simultaneous quantification of seven metabolites of polycyclic aromatic hydrocarbon in human urine was developed using online solid phase extraction-HPLC with double ternary liquid chromatography system combined with fluorescence detector. The target compounds were online concentrated on the Turboflow Cyclone solid phase extraction column at first, then transferred by the six-way valve to the Hypersil Green PAH column for separation with acetonitrile and water as mobile phase at a flow rate of 1. 0 mL/min and at 35 ℃. A single sample analysis cycle took only 20 min. Under the optimized chromatographic conditions, the method showed good linear relationship ( r≥0. 999 ) in the range of 5-2000 ng/L or 50-20000 ng/L. The LODs were 0. 5-15 ng/L, and the recoveries were 80. 7%-110. 7%. The proposed method was successfully applied in the detection of metabolites of polycyclic aromatic hydrocarbons in urine from several smokers and non-smokers. The concentrations of 2-hydroxynaphthalene, 1-hydroxynaphthalene, 2-hydroxyfluorene, 2-hydroxyphenanthrene, 4-hydroxyphenanthrene and 6-hydroxychrysene in the smokers urine were much higher than that in non-smokers.

Objective Non-invasive detection is the focus of intense research in diagnosis of acute renal allograft rejection currently. Urine protein is considered the cue to reflect the pathological changes in kidney disease. In this study, we explored the urine markers for early acute renal allograft rejection. Methods The urine protein of two patients with acute renal allograft rejection were examined by 2D gel electrophoresis and bioinformatics. We adopted pH 4-7 ready strip IPG and stained the gel with Sypro-Ruby. The digitized 2D maps of urine protein were quantitatively analyzed using 2D-analysis software packages. By analyzing the differential expressions of proteome between different time points (1, 2, 3 days before acute rejection and 7, 14, 21 days after acute rejection), 30 protein spots were selected and analyzed by MALDI-TOF-MS/MS. Results We obtained 2D gel electrophoresis maps of urine protein of the patients with acute renal allograft rejection, which are of good reproducibility and resolution. Sixteen protein spots were identified, resulting in thirteen corresponding proteins. Out of these proteins, we screened three proteins (alpha-1-antichymotrypsin, tumor rejection antigen gp96, Zn-Alpha-2-Glycoprotein) closely related to acute rejection. Conclusion The urine protein spots on 2D gel electrophoresis maps for the patients with acute renal allograft rejection were of obvious difference when detected at different time points of acute rejection. Alpha-1-antichymotrypsin, tumor rejection antigen gp96 and Zn-Alpha-2-Glycoprotein might be the candidate protein markers to diagnose acute renal allograft rejection after renal transplantation.

Objective To evaluate the safety and efficiency of domestic snare applied during retrograde percutaneous coronary intervention (PCI) for chronic total occlusion (CTO) lesions.Methods A total of 27 patients who underwent retrograde PCI for CTO and used domestic snare during the procedure were enrolled in our study from March 2012 to November 2016.Clinical data, angiographic characteristics and PCI details were collected.Clinical data, characteristics of CTO lesion, effect of the domestic snare and snaring time were retrospectively analyzed.Special complications related to the domestic snare and major adverse cardiovascular events (MACE) were also documented.Results Domestic snare was used in all the procedures, which included facillitating the micro-catheter to pass through the CTO lesions in 6 patients and assisting the RG3/rotational guide-wire externalization in all the 27 patients.Mean snaring time was 3.5±5.4 minutes.Stents were successfully implanted in 26 patients except in 1 patient who failed to receive stent implantation for severe coronary calcification.No complications including coronary dissection, fracture of guide-wire and unreleased snare happened during the procedures and no MACE occurred during hospitalization.Conclusions Domestic snare facilitates retrograde micro-catheter crossing CTO lesions and retrograde guide-wire entering the guiding catheter and externalization.It is a simple, safe and efficient method.

Objective:To study the expression of microRNA (miRNA)-4783-3p in liver cancer tissue and its effect on the proliferation and migration of liver cancer Huh-7 cells.Methods:The cBioPortal database was used to analyze the expression of miR-4783-3p in liver cancer tissues and adjacent tissues. In strict accordance with the instructions of Lipofectamine? 2000 transfection kit, miR-4783-3p overexpression mimics or overexpression control mimics were transfected into Huh-7 cells respectively, namely overexpression group and control group. The proliferation of Huh-7 cells was analyzed by CCK-8 assay, and the migration of Huh-7 cells was analyzed by cell scratch method. The targeting relationship between miR-4783-3p and insulin-like growth factor binding protein 2 ( IGFBP2) mRNA was detected by dual-luciferase reporter gene assay. RT-qPCR was used to detect the expression of IGFBP2 mRNA. Western-blotting was used to detect the expression of IGFBP2 protein and EGFR-STAT3 molecular pathway proteins. Results:The expression of miR-4783-3p in liver cancer tissues was significantly lower than that in adjacent tissues ( P<0.01). Compared with the control group, the proliferation ability of Huh-7 cells in the overexpression group was significantly decreased ( P<0.05). The scratch healing rates of Huh-7 cells in the control group and the overexpression group were (67.71±9.04)% and (29.58±10.51)%, respectively, and the scratch healing rate in the overexpression group was significantly lower ( P<0.01). miR-4783-3p targeted and bound to IGFBP2 mRNA ( P<0.01). The expression of IGFBP2 mRNA in the control and overexpression groups was 5.76±1.44 and 0.99±0.47, respectively, and miR-4783-3p negatively regulated the expression of IGFBP2 mRNA ( P<0.01). Compared with the control group, the expressions of IGFBP2 protein and EGFR-STAT3 molecular pathway protein were decreased in the overexpression group. Conclusions:miR-4783-3p is lowly expressed in liver cancer tissues. miR-4783-3p can attenuate the proliferation and invasion ability of liver cancer Huh-7 cells by inducing the low expression of IGFBP2 gene.

Objective To explore the correlation between serum fibroblast growth factor-23(FGF23),Klotho protein levels and cardiac valve calcification(CVC)in patients with maintenance hemodialysis(MHD).Methods A total of 169 MHD patients in the Blood Purification Center of Jinshan Hospital,Fudan University from January 2022 to January 2023 were selected.The patients'CVC status was examined by color Doppler echocardiography.Results There were 111 CVC cases,accounting for 65.68%.Compared with the non-CVC group,the CVC group had significantly higher levels of FGF23,β2-microglobulin,brain natriuretic peptide(BNP),age,and the proportion of male,while significantly lower levels of Klotho,predialysis creatinine,hemoglobin,plasma albumin and iPTH(P＜0.05).Pearson/Spearman correlation coefficient analysis showed that FGF23,age,β2-microglobulin and BNP were significantly positively correlated with CVC(r=0.20,0.41,0.15,0.28,0.12),while Klotho,predialysis creatinine,hemoglobin,plasma albumin and iPTH were significantly negatively correlated with CVC(r=﹣0.16,﹣0.17,﹣0.16,﹣0.17,﹣0.16).Unconditioned binary logistic regression analysis showed that FGF23,Klotho,age and β2-microglobulin were independent risk factors for CVC in MHD patients.Conclusions MHD patients are prone to CVC.Serum FGF23 protein level is significantly positively correlated with CVC,while Klotho protein level is significantly negatively correlated with CVC.FGF23,Klotho,age and β2-microglobulin are independent risk factors for CVC in MHD patients.

Objective To investigate the effect of expression of Cullin 4B (CUL4B) on the prognosis of patients after liver transplantation for hepatocellular carcinoma (HCC).Methods The retrospective case-control study was conducted.The clinicopathological data of 79 patients who underwent liver transplantation for HCC in the First Affiliated Hospital of Sun Yat-sen University between January 1,2014 and June 30,2015 were collected.The specimens of HCC tissues were collected and embedded in paraffin,and then were detected by immunohistochemistry staining.Observation indicators:(1) expression of CUL4B in HCC tissues;(2) follow-up and survival;(3) prognostic factors analysis after liver transplantation;(4) association between expression of CUL4B and recurrence and metastasis of tumor after liver transplantation.Follow-up using outpatient examination and telephone interview was performed to detect tumor recurrence or metastasis and survival up to June 2018.Measurement data with normal distribution were represented as (x)±s.The comparison between groups of count data was done using the chi-square test.The survival curve drawn using the Kaplan-Meier method,and the survival analysis was done by Log-rank test.The univariate and multivariate analysis were respectively done using the COX regression model.The association analysis was done using the Pearson test.Results (1) Expression of CUL4B in HCC tissues:immunohistochemistry staining showed that CUL4B was mainly expressed in the cytoplasm,with a powerful brownish-yellow staining.The high expression and low expression of CUL4B in HCC tissues were detected in 64 and 15 patients,respectively.(2) Follow-up and survival:79 patients were followed up for 38-56 months,with an average time of 46 months.During the follow-up,37 patients had no tumor recurrence and 42 had tumor recurrence (32 with tumor extrahepatic metastasis and 10 with intrahepatic metastasis);36 had survival and 43 died;the 1-and 3-year overall survival rates were respectively 86.84％ and 63.25％,and 1-and 3-year tumorfree survival rates were respectively 62.31％ and 51.27％.(3) Prognostic factors analysis after liver transplantation:① Results of univariate analysis showed that preoperative alpha-fetoprotein (AFP),Child-Pugh score,maximum tumour dimension,capsular invasion,intravascular tumor thrombus,Edmonson pathological grading and expression of CUL4B were related factors affecting the 3-year overall survival rate of patients after liver transplantation for HCC [Hazard Ratio (HR) =2.17,3.36,3.66,2.43,2.19,3.36,2.84,95％ confidence interval(CI):1.17-4.04,1.53-7.42,2.10-6.42,1.33-4.17,1.08-9.04,1.58-7.59,1.17-6.32,P＜ 0.05].The preoperative alpha-fetoprotein (AFP),Child-Pugh score,maximum tumour dimension,capsular invasion,intravascular tumor thrombus,Edmonson pathological grading and expression of CUL4B were related factors affecting the 3-year tumor-free survival rate of patients after liver transplantation for HCC (HR =2.06,3.72,3.16,2.36,2.83,3.21,1.69,95％CI:1.34-4.85,1.72-8.63,1.79-7.31,1.46-4.86,1.19-8.63,1.19-7.92,1.06-4.87,P＜0.05).② Results of multivariate analysis showed that maximum tumour dimension,intravascular tumor thrombus and expression of CUL4B were independent factors affecting the 3-year overall survival rate of patients after liver transplantation for HCC [Odds ratio(OR) =3.43,3.69,2.81,95％CI:1.16-6.02,1.96-9.38,1.04-9.63,P＜0.05].The maximum tumour dimension,intravascular tumor thrombus and expression of CUL4B were independent factors affecting the 3-year tumor-free survival rate of patients after liver transplantation for HCC (OR=2.25,4.72,2.74,95％C1:1.16-4.02,1.98-9.47,1.03-7.10,P＜ 0.05).The 3-year overall survival rate in patients with high-and low-expressions of CUL4B was respectively 66.7％ and 32.8％,with a statistically significant difference (x2 =5.69,P＜0.05).The 3-year tumor-free survival rate in patients with high-and low-expressions of CUL4B was respectively 73.3％ and 18.6％,with a statistically significant difference (x2 =4.63,P＜0.05).(4) Association between expression of CUL4B and recurrence and metastasis of tumor after liver transplantation:results of Pearson test showed that expression of CUL4B was significantly associated with HCC recurrence and metastasis after liver transplantation (r =0.62,P＜0.05).The further analysis showed that expression of CUL4B was significantly associated with extrahepatic metastasis after liver transplantation (r=0.84,P ＜ 0.05).Conclusion The expression of CUL4B is associated with HCC recurrence after liver transplantation,and it can be as a predictor for HCC recurrence and distant metastasis after liver transplantation.

Objective To evaluate the impact of cardiovascular risk factors on index of microvascular resistance (IMR)and coronary flow reserve (CFR) and to explore the characteristics of IMR and CFR and the relationship between IMR and angiographic features in patients with intermediate coronary stenosis and chest pain.Methods Fractional flow reserve (FFR),CFR,and IMR were measured in patients who underwent invasive coronary angiography with 40％-70％ stenosis by visual assessment.All patients with FFR＞0.75 were enrolled and grouped with the cut-off points of IMR≥25 and CFR≤2.0.Patients with IMR≥25 were group H,including two sub-groups (high IMR-low CFR,group H1 and high IMR-high CFR,group H2),while those with IMR＜25 were group N.The thrombolysis in myocardial infarction (TIMI) frame were counted.Results A total of 34 patients with FFR＞0.75 were enrolled with 61.8％(21 cases) of males and 38.2％ (13 cases) of females.The mean age was (57.3±8.1) years old.High IMR accounted for 47.1％ of all cases.There was significant difference between group H and N in TIMI frame (33.0 vs.20.8,P=0.031).There were significant differences between group H1 and H2 in homocysteine (17.8 μmol/L vs.12.0 μmol/L,P=0.005) and IMRcorr (58.0 vs.36.1,P=0.002).IMRcorrwas correlated to TIMI frame (r=0.40,P=0.012) for all cases.The sensitivity and specificity of inferring IMR≥35.3 by TIMI frame were 0.75 and 0.65 (P=0.049) with TIMI frame over 40.5.Conclusions High IMR may be one of the reasons for chest pain in patients with intermediate coronary stenosis.There is no correlation between vascular risk factors and IMR or CFR,while there is positive correlation between TIMI frame and IMR.The specificity is 65％ for inferring IMR rise with TIMI frame over 40.5.

Kidney organoids are induced and differentiated from human pluripotent stem cells(PSCs)or adult stem cells(ASCs)derived from tissue sources,primarily composed of nephron struc-tures.However,due to the absence of a supporting vascular network,kidney organoids often exhibit immature tissue structures and limited growth.Therefore,vascularization in kidney organoids remains a pressing challenge in this field.Currently,the methods such as transplantation into immunodeficient animals,alterations in induced differentiation protocols,utilization of microfluidic chips,and manipula-tion of extracellular matrix and oxygen concentrations may facilitate vascularization of kidney organoids,which provides a new perspective for the scientific study and clinical application of kidney organoids.