Objective To investigate the role of tet methylcytosine dioxygenase 2 (TET2) in the regulation of transforming growth factor-β1 (TGF-β1) expression in human glomerular mesangial cells induced by high glucose.Methods Cultured human glomerular mesangial cells were divided into normal control group (5.5 mmol/L glucose) and high glucose group (30.0 mmol/L glucose) which was cultured for 12 h to 72 h.The gene expression of TET2 in mesangial cells were inhibited by small molecule chemical called SC1,and which were divided into high glucose group (30.0 mmol/L glucose+ DMEM),DMSO group (30.0 mmol/L glucose+0.1％DMSO) and SC1 group (30.0 mmol/L glucose+3 μmol/L SC1).The mRNA and protein expression of TGF-β1,TET1 to 3 and α-smooth muscle actin (α-SMA) was detected by quantitative real-time PCR and Western blotting.Methylation of CpG islands in the regulation region of TGF-β1 was detected by bisulfite sequencing PCR (BSP).The activity of mesangial cell proliferation was assessed by colorimetry of thiazolyl blue (MTT).Results Compared with normal control group,the mRNA and protein expression of TET2 in mesangial cells induced by high glucose was increased significantly in a time-dependent manner (all P ＜ 0.05),but the expression of TET1 and TET3 was not affected.Meanwhile methylation rate of 4 CG sites from 24 h to 72 h were decreased in the first exon of TGF-β1 (P ＜ 0.01),but not in the promoter.Compared with high glucose group,when the expression of TET2 was inhibited by SC1,the methylation rate of TGF-β1 was recovered evidently (P ＜ 0.05),the mRNA and protein expression of TGF-β1 and α-SMA was suppressed,and the proliferation of mesangial cells was decreased (all P ＜ 0.05).Conclusions Demethylation of the CpG island mediated by TET2 may play an important role in the expression of TGF-β1 and mesangial cell phenotype transformation induced by high glucose.

Objective: To apply optical coherence tomography angiography (OCTA) to evaluate the clinical effects of intravitreal injection of antivascular endothelial growth factor (VEGF) agents on wet age-related macular degeneration (AMD). Methods: The 31 eyes of 31 wet AMD patients in the Department of Ophthalmology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine from April 2018 to December 2018 were included. These patients received monthly intravitreal injection of anti-VEGF agents for three consecutive months. The best-corrected visual acuity (BCVA) before and 1, 2 and 3 months after first injection was compared. The macular fovea thickness and choroidal neovascularization (CNV) area at different time points before and after treatment were detected by OCTA and compared. Results: The baseline BCVA (logarithm of the minimum angle of resolution) of the included patients was 0.93±0.43, and the BCVA was significantly improved after treatment (P<0.05). The results of OCTA showed that before treatment the inner limiting membrane-retinal pigment epithealium thickness and inner limiting membrane-retinal pigment epithelial fit thickness were (329.03±110.73) μm and (468.84±209.50) μm, respectively, and they both decreased significantly after treatment (P<0.05). The CNV area before treatment was (4.78±3.24) mm2, and it decreased gradually with time after treatment (P<0.05). Conclusion: Intravitreal injection of anti-VEGF agents can reduce macular edema and inhibit CNV in the wet AMD patients. OCTA can be used to evaluate the efficacy of intravitreal injection of anti-VEGF agents in the treatment of wet AMD.

Objective To investigate the value of delayed 18F-FDG PET/CT with oral intake small dosage diuretics for diagnosing urogenital cancers.Methods Patients with suspected urogenital system cancers were divided into routine dosage diuretic group (n=12) and small dosage diuretics group (n=35).All patients underwent whole-body PET/CT followed by delayed scanning after oral 40 mg or 20 mg Furosemide respectively.The urine maximum standard uptake value (SUVmax) and T/U (the ratio of urine and lesion SUVmax) before and after diuresis were compared respectively.Diagnostic efficacy for malignant urogenital system cancers of small dosage group was calculated.Results The urine SUVmax and T/U were statistically different between routine whole body and delayed scans in both groups (P＜0.05).SUVmax and T/U of routine and delayed scans had no statistical differences between the two groups (P＞0.05).In small dosage group,the sensitivity,specificity,positive predictive value,negative predictive value and accuracy of delayed imaging and routine imaging was 96.77％ (30/31)and 61.29％ (19/31),75.00％ (3/4) and 50.00％ (2/4),96.77％ (30/31) and 90.48％ (19/21),75.00％ (3/4)and 14.29％ (2/14),94.29％ (33/35) and 60.00％ (21/35),respectively.The sensitivity and accuracy were statistically different between routine and delayed imaging (P＜0.001).Conclusion Delayed PET/CT imaging with oral small dosage Furosemide has the same efficacy as PET/CT using routine dosage diuretics,which is useful for diagnosing urogenital cancers.

Polyhydroxyalkanoates (PHA) is a family of microbially synthesized polyesters consisting of various 3-hydroxyalkanoate monomers. Aeromonas hydrophila 4AK4 could be able to synthesize PHA copolymer consisting of 3-hydroxybutyrate (3-HB) and 3-hydroxyhexanoate (3-HHx). No data has been reported about the ability to synthesize the PHA with other monomers in A. hydrophila. In this study, propionic acid, valeric acid, heptanoic acid, nonanoic acid and undecanoic acid were used together with gluconate to find out whether A. hydrophila 4AK4 could synthesize the PHA consisting of odd carbon atom number monomers. The result showed that A. hydrophila 4AK4 could not growth when supplied with propionic acid, valeric acid, heptanoic acid and nonanoic acid and only undecanoic acid could be used to synthesize PHA. Wild type and recombinant A. hydrophila 4AK4 harboring phaA (beta-ketothiolase) and phaB (acetoacetyl-CoA reductase) were cultivated with undecanoic acid and glucose or undecanoic acid and gluconate served as carbon sources. PHA consisting of 3-HB and 3-hydroxyvalerate (3-HV) could be produced by both wild type and recombinant A. hydrophila 4AK4 and the latter could produce PHA with more 3-HB monomer. When the ratio of glucose or gluconate to undecanoic acid was 1:1, the cell dry weight (CDW) of A. hydrophila 4AK4 reached 1.14 g/L and PHA content was 60% of the CDW after cultivation for 24 h. When lauric acid and undecanoic acid were served as co-substrate, A. hydrophila 4AK4 could produce copolyester consisting of 3-HB, 3-HV and 3-HHx. Along with the increase of undecanoic acid proportion in the mixed carbon source, the 3-HV content of copolymer was increased while the 3-HB and 3-HHx content were decreased. In all cases, the CDW decreased along with the increase of undecanoic acid concentration, which indicated that undecanoic acid was not very good for A. hydrophila 4AK4 growth.
Aeromonas hydrophila ; metabolism ; Fatty Acids ; metabolism ; Glucose ; metabolism ; Lauric Acids ; metabolism ; Pentanoic Acids ; metabolism ; Polyhydroxyalkanoates ; biosynthesis

OBJECTIVE: To determine whether diffusion kurtosis imaging (DKI) is effective in monitoring tumor response to neoadjuvant chemotherapy in patients with osteosarcoma. MATERIALS AND METHODS: Twenty-nine osteosarcoma patients (20 men and 9 women; mean age, 17.6 ± 7.8 years) who had undergone magnetic resonance imaging (MRI) and DKI before and after neoadjuvant chemotherapy were included. Tumor volume, apparent diffusion coefficient (ADC), mean diffusivity (MD), mean kurtosis (MK), and change ratio (ΔX) between pre- and post-treatment were calculated. Based on histologic response, the patients were divided into those with good response (≥ 90% necrosis, n = 12) and those with poor response (< 90% necrosis, n = 17). Several MRI parameters between the groups were compared using Student's t test. The correlation between image indexes and tumor necrosis was determined using Pearson's correlation, and diagnostic performance was compared using receiver operating characteristic curves. RESULTS: In good responders, MDpost, ADCpost, and MKpost values were significantly higher than in poor responders (p < 0.001, p < 0.001, and p = 0.042, respectively). The ΔMD and ΔADC were also significantly higher in good responders than in poor responders (p < 0.001 and p = 0.01, respectively). However, no significant difference was observed in ΔMK (p = 0.092). MDpost and ΔMD showed high correlations with tumor necrosis rate (r = 0.669 and r = 0.622, respectively), and MDpost had higher diagnostic performance than ADCpost (p = 0.037) and MKpost (p = 0.011). Similarly, ΔMD also showed higher diagnostic performance than ΔADC (p = 0.033) and ΔMK (p = 0.037). CONCLUSION: MD is a promising biomarker for monitoring tumor response to preoperative chemotherapy in patients with osteosarcoma.
Bone Neoplasms ; Diffusion ; Drug Therapy ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Necrosis ; Osteosarcoma ; ROC Curve ; Tumor Burden

Aged ; Aged, 80 and over ; External Fixators ; Female ; Humans ; Male ; Manipulation, Orthopedic ; Middle Aged ; Radius Fractures ; therapy ; Splints

BACKGROUNDThe induced expression of heme oxygenase-1 (HO-1) in donor islets improves allograft survival. Cobalt protoporphyrin (CoPP) could significantly enhance the expression of HO-1 mRNA and protein in rat islet safely. Our work was to study how to protect pancreatic islet xenograft by CoPP-induction.

METHODSIslet xenografts treated with CoPP-induction and CoPP + Zinc protoporphyrin (ZnPP) in vitro and in vivo were randomly transplanted into murine subrenal capsule; then the graft survival time was compared by blood glucose level and pathological examination and meanwhile the interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), interleukin 10 (IL-10) and IL-1β level in serum and their mRNA and HO-1 mRNA and protein expression were examined.

RESULTSIslets with CoPP-induction under low- and high-glucose stimulation exhibited much higher insulin secretion compared with other three groups. CoPP-induction could increase higher expression of HO-1 (mRNA: 3.33- and 76.09-fold in vitro and in vivo; protein: 2.85- and 58.72-fold). The normoglycemia time in induction groups ((14.63 ± 1.19) and (16.88 ± 1.64) days) was significantly longer. The pathological examination showed less lymphocyte infiltration in induction groups. The IL-10 level and its mRNA in induction groups were significantly higher.

CONCLUSIONSThe HO-1 induced by CoPP would significantly improve function, prolong normoglycemia time and reduce lymphocyte infiltration. Meanwhile CoPP-induction in vivo had more beneficial effects than in vitro. Its mechanism could be related to immune-modulation of IL-10.

Animals ; Blotting, Western ; Graft Survival ; Heme Oxygenase-1 ; genetics ; metabolism ; Interleukin-10 ; blood ; Islets of Langerhans ; enzymology ; metabolism ; Islets of Langerhans Transplantation ; Male ; Mice ; Mice, Inbred BALB C ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Heterologous

OBJECTIVEVasoactive intestinal peptide (VIP) is a neuro-peptide that can modulate immunity. Previous studies indicated that VIP can attenuate the deleterious consequences of severe sepsis and septic shock by regulating production of inflammatory cytokines in immune activated cells. The signaling induced by bacterial components occurs primarily through Toll like receptors (TLRs). TLRs have been recognized to play a key role in pathogen recognition and innate immunity. It was convincingly demonstrated that lung is one of early suffered disaster organ and may trigger multiple organ dysfunction syndrome in sepsis. The present study was conducted to investigate the effects of VIP on TLR2/4 mRNA expressions on acute lung injury of endotoxic shock induced by lipopolysaccharide (LPS) in rat.

METHODForty Sprague-Dawley rats were randomly divided into 3 groups, i.e., LPS shock group (n = 16), LPS + VIP group (n = 16), and control group (n = 8). LPS shock model was established by LPS (E. coli O(55)B(5) 10 mg/kg) with tail intravenous injection. The rats in LPS + VIP group were given a bolus of 5 nmol VIP intravenous injection follow by LPS. The rats in control group were given normal saline. The rats were sacrificed at 6 h, 24 h after being injected. The lung tissues were collected. The TLR2 mRNA and TLR4 mRNA expressions were detected by RT-PCR from the lung tissues. Pathological changes of the lungs were observed by light microscope and electron microscope 24 h after LPS injection.

RESULT(1) Lung histopathology: the alveolar space was full with leukocyte, necrotic cells, segmental hemorrhage and protein effusion. Partial alveolar space was enlarged, lung interstitial edema were observed in LPS shock group. However, pathological changes of LPS + VIP group were milder than those in LPS shock group. (2) The expressions of TLR2 mRNA and TLR4 mRNA were significantly higher in LPS shock group compared with those of the control group (F = 16.638, P = 0.000; t = 5.876, P = 0.000), TLR2 mRNA and TLR4 mRNA expression on 24 h was down-regulated in LPS + VIP shock subgroup than those in LPS shock subgroup (F = 16.676, P = 0.000; t = 3.946, P < 0.001).

CONCLUSIONExpressions of TLR2 mRNA and TLR4 mRNA were up-regulated on LPS induced lung injury in rats. VIP mitigated lung injury induced by LPS, which may be related to TLR2 mRNA and TLR4 mRNA down-regulation of expression. The effect of VIP may suggest a protective mechanism in sepsis. VIP may play a potential protective role in severe infection.

Acute Lung Injury ; chemically induced ; metabolism ; pathology ; Animals ; Down-Regulation ; Lipopolysaccharides ; Lung ; metabolism ; pathology ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Up-Regulation ; Vasoactive Intestinal Peptide ; pharmacology

[Objective]To radiolabel the PSMA aptamer A10-3.2 with 99mTc , and explore its biological characteristics in vivo and in vitro.[Methods]Using Succinimidyl 6-hydrazinonicotinate hydrochloride (SHNH) as the bifunctional chelating agent to label aptamer A10-3.2 with 99mTc, then tested for the stability in vitro, the specific uptake by prostate cancer LNCaP cells (PSMA+) , the characteristics of SPECT/CT imaging and biodistribution in LNCaP tumor-bearing NOD/SCID mice.[Results]The labeling rate and radiochemical purity of the products (99mTc-SHNH-A10-3.2) are(71.31 ± 6.78)% and 97.03%,respectively. 99mTc-SHNH-A10-3.2 had obvious target specificity for PSMA positive prostate cancer LNCaP cells, its uptake rate was significantly higher than PSMA nega?tive PC-3 cells (P<0.01). And in tumor-bearing mice, the tumor has a certain uptake and a high ratio of the tumor tissue to the mus?cle.[Conclusion]This study successfully constructed 99mTc-labeled PSMA-targeted aptamer A10-3.2, which has a good stability and targeting in vivo and in vitro, has a high tumor tissue/muscle ratio in tumor-bearing mice, which show that it may be a potential target?ed molecular imaging agent for prostate cancer.

OBJECTIVETo select and identify the target genes related to oral squamous cell carcinoma (OSCC) and provide target genes for designing oligo-nucleotide functional microarray of OSCC.

METHODSGenes possibly related to oral squamous cell carcinoma were selected from the 5 years' published data of differently expressed profiles with microarray testing in OSCC. Then mRNA expression of selected genes were evaluated by real time quantitative polymerase chain reaction (RT-PCR) in 22 cases of OSCC, including tumor tissues and paried normal mucosas and quantified according to an internal control GAPDH.

RESULTSEight genes were tested. The overexpression of SPARC, PDGF-A, SERPINE1, TGF-beta(1) and VEGF-C genes were measured in 16, 18, 16, 20, 18 cases of tumor specimens, respectively. The expression of CK15 gene was lower than that of its normal tissue. There were overexpression of CCND1, BIRC3 in tumor tissues, but there was no significant difference of CCND1 and BIRC3 expression between tumor tissue and normal tissue (P > 0.05).

CONCLUSIONSSPARC, PDGF-A, SERPINE1, TGF-beta(1), VEGF-C and CK15 genes were closely related to tumor progress of OSCC. They can be used as the target genes for designing oligo-nucleotide functional microarray of OSCC.

Adult ; Aged ; Biomarkers, Tumor ; genetics ; Carcinoma, Squamous Cell ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mouth Neoplasms ; genetics ; Oligonucleotide Array Sequence Analysis ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods