Objective To investigate the effect of IGF-1R inhibitor AG1024 on the esophageal cancer xenografts and the underlying mechanisms.Methods The mouse model was established by injecting EC9706 cells subcutaneous in nude mice.When the tumors were 100 mm3 in size,the mice were divided into 4 groups randomly withcontrol group with no treatment; irradiation group with 8 Gy 6 MV Xrays at 1 and 8 d each time; AG1024-treatment group with 30 μg/(kg-d) AG1024 injected intraperitoneally (ip) 5 times a week for two weeks ; combination group:receiving both 30 μg/(kg· d)-1 AG1024 ip and irradiation of 8 Gy X-rays.The diameters of the tumors were measured every 3 days.The mice were sacrificed and the weights of tumors were measured at 15 d after treatment.Tumor inhibition rate was calculated.The cell cycle was examined by flow cytometry.The expression of cell cycle protein D1 (CyclinDl) of the tumors were detected by immunohistochemical staining.Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) assay was used to detect the cell apoptosis in the tumor tissue.Results The tumor weight in the irradiation group,AG1024-treatment group and combination group were significantly decreased compared with the control group,and the inhibition rate were 39.16％,18.73％,57.04％,respectively (F =13.566,P ＜ 0.05).After treatment with AG1024 and irradiation,tumor tissue cells were significantly accumulatedin the G0/G1 and G2/M phases and decreased in S phase compared to the irradiation group (t =-6.654,-16.738,12.871,P ＜ 0.05).The CyclinD1 expression of the combination group was significantly decreased compared with the control group.In the combination group,the apoptotic cells were detected by TUNEL assay.Conclusions IGF-1R inhibitor AG1024 could change the cell cycle and induce cell apoptosis,which might result in the enhancement of radiosensitivity on the esophageal cancer xenografts.

Objective To explore the association between SLC2A9,SLC17A3,ABCG2 single nucleotide polymorphisms and gout susceptibility in Quanzhou.Methods One hundred and fifty-four cases of gout patients and 160 healthy controls were selected,single nucleotide polymorphisms (SNP) of SLC2A9 SLC17A3,ABCG2 with tri-primer polymerase chain reaction (PCR) were tested and the relation between different genotypes and primary gout prevalence were analyzed.Results High risk genotype frequency of rs16890979 was 93.5％ and 70.0％ in patients and healthy people,respectively (the difference of genotype frequency between the two groups was statistically significant (x2=55.377,P＜0.01).High risk allele frequency was 79.9％ and 48.4％ in patients and healthy people,respectively (allele frequency in different population was statistically significant,x2=67.128,P＜0.01).High risk genotype frequency of rs2231142 was 68.8％ and 38.7％ in patients and healthy people,respectively (the difference of the genotype frequency was statistically significant,x2=29.129,P＜0.01);High risk allele frequency was 43.5％ and 23.4％ in patients and healthy people,respectively (the difference of allele frequency was statistically significant,x2=28.468,P＜0.01) ; rs1165205was a protective SNP,low risk genotype frequency was 42.2％ and 45.6％ in patients and healthy people,respectively (the difference of genotype frequency was statistically significant,x2=0.373,P=0.571); High risk allele frequency was 26.0％ and 28.1％ in patients and healthy people,respectively (the difference of allele frequency was not statistically significant,x2=0.270,P=0.364).Conclusion SNP loci rs16890979 of SLC2A9 gene and rs2231142 of ABCG2 gene can be used as genetic markers for primary gout susceptibility in the Quanzhou area,but SNP loci rs1165205 of SLC17A3 gene has little correlation with the prevalence of primary gout in Quanzhou residents.

Objective: To analyze the clinical characteristics of hypophosphatemic osteomalacia induced by adefovir dipivoxil (ADV) in order to improve the understanding of the disease.Methods: A retrospective analysis was performed according to the medical records of 11 cases of ADV-induced hypophosphatemic osteomalacia.The medical history, laboratory indicators (ALT, AST, ALB, SCr, UA, blood glucose, blood pH, BE), bone metabolic markers (25OHD3, PTH, tP1NP, β-CTX, OC), urine indicators (urine pH, 24h urine Ca, 24h urine P, 24h urine Pro, urine Scr), DXA and skeleton ECT signs of the patients with hypophosphatemic osteomalacia induced by ADV were analyzed, and the symptoms, blood P, AKP level and urine routines were followed up after 1-month withdrawal and in July, 2016, respectively.Results: The mean ADV administration time of the 11 patients was (5.7±1.2) years, and the bone pain time was (2.2±0.6) years.The serum P was (0.45±0.99)mmol·L-1, 24h urine P was (17.9±4.8)mmol, AKP was (248±107)IU·L-1,the concentration threshold of renal phosphate was(0.31±0.10)mmol·L-1.After the one-month withdrawal of ADV, the bone pain in the patients were all relieved, and with the phosphorus supplement, the level of serum phosphorus was increased.In July of 2016, the average withdrawal time of ADV was (18.3±10.7) months, the serum phosphorus significantly increased and AKP significantly decreased when compared with that on the admission and 1 month after the ADV withdrawal (P<0.05), and the serum phosphorus of 2 patients returned to normal with the recovery rate of 20% (2/10).The regression analysis showed that the influencing factors on serum phosphorus on the admission were renal concentration threshold of phosphate and tP1NP (P<0.05);the influencing factor on serum phosphorus on the last follow-up was bone mineral density at the admission (P<0.05).Conclusion: Hypophosphatemic osteomalacia is a potential side effect of ADV, and ADV-induced renal injury is not completely reversible, which should be paid more attention in clinical work.

Objective To prepare 18F-5-fluoro-N-(2-(diethylamino) ethyl) picolinamide (18F-5-FPN) and evaluate its binding affinity with melanin.Methods 5-bromo-N-(2-(diethylamino) ethyl)picolinamide (precursor) was synthesized and the structure was characterized by ultraviolet (UV),nuclear magnetic resonance (NMR) and mass spectrometry.18F-5-FPN was prepared with FX-XN module through nucleophilic substitution reaction.The product was purified and identified by HPLC.The binding specificity of 18F-5-FPN with melanin was demonstrated by in vitro study of cellular uptake,and in vivo static PET imaging of pigmented B16F10 and amelanotic A375m allografts.Biodistribution study was performed to evaluate the pharmacokinetics of 18F-5-FPN in vivo.Two-sample t test was used for data analysis.Results The structure of precursor was characterized by UV,1H NMR,13C NMR and mass spectrometry.18 F-5-FPN was successfully prepared with radiochemical yield of 5％-8％,radiochemical purity ＞95％ and the specific activity of 100-120 GBq/μmol.The cell uptake study showed that the uptakes of 18F-5-FPN in B16F10 cells at 30,60 and 120 min ((9.80±0.46) ％,(10.34±0.32) ％,(7.27±0.26)％) were significantly higher than those in A375m cells ((1.36±0.14)％,(1.75±0.12)％,(1.54±0.09)％;t =30.3,46.8,38.4,all P＜0.05),and the optimal uptakes were observed at 60 min for both cells.In static PET imaging,the tumors in B16F10-bearing mice were clearly visible with the uptake value of (18.20±3.21) ％ID/g and the T/B ratio of 19.17±10.03 at 1 h postinjection,while no tumor uptake was seen in A375m-bearing mice.The main clearance pathway of 18F-5-FPN was the renal system,which cleared the unbound tracer rapidly.Conclusions 18F-5-FPN can specifically target the melanin in vitro and in vivo with favorable pharmacokinetics and good T/B ratio.18F-5-FPN may be an ideal molecular probe for diagnosis of malignant melanoma.

Objective·To evaluate the relationship between body mass index(BMI)and chronic metabolic diseases.Methods·The elderly(≥60 years old)who were underwent physical examination in the Physical Examination Center of Renji Hospital,Shanghai Jiao Tong University School of Medicine from 2014 to 2021 were studied.Their results of biochemical indicators were collected.Their height,body weight,and blood pressure were measured by trained nurses.The history of chronic metabolic diseases was collected by self-reported questionnaire.Systolic blood pressure≥140 mmHg(1 mmHg=0.133 kPa),diastolic blood pressure≥90 mmHg,or self-reported hypertension history was defined as hypertension.Fasting blood glucose≥7.0 mmol/L or self-reported history of diabetes was defined as diabetes.Total cholesterol≥6.2 mmol/L,triglyceride≥2.3 mmol/L,or self-reported history of dyslipidemia was defined as dyslipidemia.The relationship between BMI and hypertension,diabetes,and dyslipidemia was evaluated by using receiver operator characteristic(ROC)curve analysis and binary logistic regression.Results·Data of 59 083 subjects were collected[30 807 men and 28 276 women,average age:(67.9±6.3)years old].The prevalence of hypertension,diabetes and dyslipidemia was 76.5%(45 219/59 083),24.1%(14 225/59 083)and 50.0%(29 544/59 083),respectively.Compared to the elderly people aged 60?74 years,those aged 75 years and above had a higher proportion of hypertension and diabetes,and a lower proportion of dyslipidemia and no metabolic abnormalities.With ROC analysis,the BMI cut-off values for hypertension,diabetes,and dyslipidemia were 24.3,23.9,and 23.9 kg/m2.The BMI cut-off values for hypertension and diabetes in elderly men were similar to those in elderly women(for hypertension:24.3 kg/m2 in elderly men vs 24.2 kg/m2 in elderly women;for diabetes:24.0 kg/m2 in elderly men vs 23.7 kg/m2 in elderly women);however,BMI cut-off value for dyslipidemia was obviously higher in elderly men than that in elderly women(24.0 kg/m2 in elderly men vs 22.5 kg/m2 in elderly women).The BMI cut-off value for chronic metabolic diseases was higher in the elderly people aged 60?74 years than that in the elderly people aged 75 years and above(24.2?24.7 kg/m2 vs 22.9?23.8 kg/m2).Conclusion·Elderly people aged 60?74 years should maintain the BMI below 24.0 kg/m2,while those aged 75 years and above should aim for the BMI below 23.0 kg/m2,so as to reduce the risk of chronic metabolic diseases.

With persistent advancement of surgical instruments, methods and techniques, clinical efficacy of liver transplantation has been steadily enhanced. However, the length of anhepatic phase is still an important factor affecting the efficacy of liver transplantation. Rat is one of the major animal models for liver transplantation-related basic research. In this article, multiple approaches for prolonging the anhepatic phase and shortening the operation time during anhepatic phase in rat liver transplantation were reviewed, which consisted of sevoflurane inhalation anesthesia, intravenous infusion via jugular vein indwelling needle, clamping of the abdominal aorta before anhepatic phase, injection of normal saline into portal vein before anhepatic phase, subcutaneous transposition of the spleen, electrocoagulation of hepatic esophageal artery, magnetic ring anastomosis of the superior and inferior hepatic vena cava, cannula anastomosis of the superior and inferior hepatic vena cava, stent anastomosis of the superior and inferior hepatic vena cava, rapid connection device and cannula of portal vein, and ring-shaped cannula of hepatic tissue-preserving inferior hepatic vena cava, aiming to add evidence for prolonging the duration of anhepatic phase, improving the operation efficiency during anhepatic phase and elevating the success rate of rat liver transplantation.

Objective:To investigate the value of serum miR-143 level combined with MRI in predicting the early response to concurrent chemoradiotherapy (CCRT) in cervical cancer.Methods:A total of 85 patients with pathologically confirmed cervical cancer underwent conventional MRI, intravoxel incoherent motion diffusion-weighted imaging (IVIM-DWI), and dynamic contrast-enhanced MRI (DCE-MRI) before CCRT. The biopsy tissues and serum samples were collected. The differential expression of miRNA in the biopsy tissues was determined by microarray chip. The expression level of miR-143 in the serum samples was analyzed by qRT-PCR. All patients were divided into the non-residual and residual tumor groups according to post-treatment MRI. Pre-treatment clinical factors, MRI parameters and miR-143 between two groups were statistically analyzed by the univariate and multivariate analyses. The optimal thresholds and predictive performance for post-treatment incidence of residual tumors were estimated by drawing the ROC curve.Results:At one month after CCRT, there were 52 patients in the non-residual tumor group and 33 patients in the residual tumor group. In the residual tumor group, pre-treatment FIGO staging, apparent diffusion coefficient (ADC), D and V e were significantly higher (all P<0.05), whereas K trans value was significantly lower ( P<0.001) when compared to those in the non-residual tumor group. The miRNA array analysis showed that there were 16 miRNAs with differential expression levels between two groups (all P<0.05). Among them, the increase of miR-143 was the most significant in the residual tumor group. Compared with the residual tumor group, the expression level of serum miR-143 was significantly down-regulated in the non-residual tumor group ( P=0.002). Compared with the SiHa cells, the expression level of miR-143 in the SiHa-R cells was significantly up-regulated ( P<0.05). Multivariate analysis showed that only miR-143, D, K trans and V e were the independent prognostic factors. The combination of multi-parametric MRI and miR-143 exhibited the highest predictive performance (AUC=0.975), with a sensitivity of 84.8% and a specificity of 96.2%. Conclusion:The combination of multi-parametric MRI with miR-143 further improves the predictive performance for residual tumors after CCRT, which contributes to the personalized treatment of cervical cancer.

The construction of efficient and stable Lactobacillus expression vector is critical for strain improvement and development of customized strains. In this study, four endogenous plasmids were isolated from Lacticaseibacillus paracasei ZY-1 and subjected to functional analysis. The Escherichia coli-Lactobacillus shuttle vectors pLPZ3N and pLPZ4N were constructed by combining the replicon rep from pLPZ3 or pLPZ4, the chloramphenicol acetyltransferase gene cat from pNZ5319 and the replicon ori from pUC19. Moreover, the expression vectors pLPZ3E and pLPZ4E with the promoter P_ldh3 of lactic acid dehydrogenase and the mCherry red fluorescent protein as a reporter gene were obtained. The size of pLPZ3 and pLPZ4 were 6 289 bp and 5 087 bp, respectively, and its GC content, 40.94% and 39.51%, were similar. Both shuttle vectors were successfully transformed into Lacticaseibacillus, and the transformation efficiency of pLPZ4N (5.23×10²-8.93×10² CFU/μg) was slightly higher than that of pLPZ3N. Furthermore, the mCherry fluorescent protein was successfully expressed after transforming the expression plasmids pLPZ3E and pLPZ4E into L. paracasei S-NB. The β-galactosidase activity of the recombinant strain obtained from the plasmid pLPZ4E-lacG constructed with P_ldh3 as promoter was higher than that of the wild-type strain. The construction of shuttle vectors and expression vectors provide novel molecular tools for the genetic engineering of Lacticaseibacillus strains.
Lacticaseibacillus ; Lacticaseibacillus paracasei ; Plasmids/genetics* ; Genetic Vectors/genetics* ; Lactobacillus/genetics* ; Escherichia coli/genetics*

ObjectiveBased on network pharmacology and molecular docking techniques， the mechanism of essential oil from Chimonanthus nitens leaves （CLO） in the treatment of acute lung injury （ALI） was predicted， and a rat model of ALI was established to verify the mechanism of CLO. MethodThe composition of CLO was determined by gas chromatography-mass spectrometry （GC-MS）. The component targets were obtained from PharmMapper and SwissTargetPrediction databases， ALI-related targets were obtained from GeneCards， Online Mendelian Inheritance in Man （OMIM） and DisGeNET， cross-over analysis with differential expressed genes （DEGs） of ALI obtained from Gene Expression Omnibus （GEO） on the Venny 2.1.0 platform yielded potential anti-ALI targets of CLO. Protein-protein interaction （PPI） analysis of potential targets was carried out by STRING 11.5. The tissue expression profiles of potential targets were obtained from the National Center for Biotechnology Information （NCBI） and the target tissue distribution maps were constructed. Potential targets were analyzed for Gene Ontology （GO） function and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment by RStudio 4.0.0 software. Composition-target-pathway network was constructed by Cytoscape 3.9.1 software， and key components and pathways were screened out and verified by molecular docking. ALI model was established by lipopolysaccharide （LPS） induction， levels of interleukin （IL）-6 and tumor necrosis factor （TNF）-α in serum of rats were measured， the expression levels of IL-17 protein in the lung tissue of ALI rats were analyzed by immunohistochemistry. ResultA total of 19 components of CLO were identified by GC-MS， and 18 potential targets were obtained by target screening. After PPI analysis， 15 target proteins with interaction relationship were obtained， further analysis showed that they were highly expressed in lung and thymus. The network diagram of component-target-pathway was analyzed to obtain the key components， including bornyl acetate， linalool， elemol， geranyl isobutyrate， and the core targets of matrix metalloproteinase 13 （MMP13）， S100 calcium binding protein A9 （S100A9）， spleen tyrosine kinase （SYK）， as well as the main signaling pathways， such as IL-17 and TNF. The results of molecular docking showed that the key components were stably bound to MMP13 and S100A9 of IL-17 signaling pathway. The results of pharmacological experiment confirmed that CLO could significantly inhibit the expression of IL-6 and TNF-α in serum of ALI rats， and decrease the expression of IL-17 protein in lung tissue of ALI rats. ConclusionCLO can achieve the therapeutic effect on ALI and protect lung tissue， the mechanism may be related to the decrease of the expression of IL-6 and TNF-α in serum and the inhibition of the activation of IL-17 signaling pathway in lung tissue of ALI rats.

BACKGROUND: Breast cancer patients who are positive for hormone receptor typically exhibit a favorable prognosis. It is controversial whether chemotherapy is necessary for them after surgery. Our study aimed to establish a multigene model to predict the relapse of hormone receptor-positive early-stage Chinese breast cancer after surgery and direct individualized application of chemotherapy in breast cancer patients after surgery. METHODS: In this study, differentially expressed genes (DEGs) were identified between relapse and nonrelapse breast cancer groups based on RNA sequencing. Gene set enrichment analysis (GSEA) was performed to identify potential relapse-relevant pathways. CIBERSORT and Microenvironment Cell Populations-counter algorithms were used to analyze immune infiltration. The least absolute shrinkage and selection operator (LASSO) regression, log-rank tests, and multiple Cox regression were performed to identify prognostic signatures. A predictive model was developed and validated based on Kaplan-Meier analysis, receiver operating characteristic curve (ROC). RESULTS: A total of 234 out of 487 patients were enrolled in this study, and 1588 DEGs were identified between the relapse and nonrelapse groups. GSEA results showed that immune-related pathways were enriched in the nonrelapse group, whereas cell cycle- and metabolism-relevant pathways were enriched in the relapse group. A predictive model was developed using three genes ( CKMT1B , SMR3B , and OR11M1P ) generated from the LASSO regression. The model stratified breast cancer patients into high- and low-risk subgroups with significantly different prognostic statuses, and our model was independent of other clinical factors. Time-dependent ROC showed high predictive performance of the model. CONCLUSIONS A multigene model was established from RNA-sequencing data to direct risk classification and predict relapse of hormone receptor-positive breast cancer in Chinese patients. Utilization of the model could provide individualized evaluation of chemotherapy after surgery for breast cancer patients.
Humans ; Female ; Breast Neoplasms/genetics* ; East Asian People ; Neoplasm Recurrence, Local/genetics* ; Breast ; Algorithms ; Chronic Disease ; Prognosis ; Tumor Microenvironment